Adenosine Deaminase Acting on RNA 1 Associates with Orf Virus OV20.0 and Enhances Viral Replication.
Orf virus (ORFV) infects sheep and goats and is also an important zoonotic pathogen. The viral protein OV20.0 has been shown to suppress innate immunity by targeting the double-stranded RNA (dsRNA)-activated protein kinase (PKR) by multiple mechanisms. These mechanisms include a direct interaction with PKR and binding with two PKR activators, dsRNA and the cellular PKR activator (PACT), which ultimately leads to the inhibition of PKR activation. In the present study, we identified a novel association between OV20.0 and adenosine deaminase acting on RNA 1 (ADAR1). OV20.0 bound directly to the dsRNA binding domains (RBDs) of ADAR1 in the absence of dsRNA. Additionally, OV20.0 preferentially interacted with RBD1 of ADAR1, which was essential for its dsRNA binding ability and for the homodimerization that is critical for intact adenosine-to-inosine (A-to-I)-editing activity. Finally, the association with OV20.0 suppressed the A-to-I-editing ability of ADAR1, while ADAR1 played a proviral role during ORFV infection by inhibiting PKR phosphorylation. These observations revealed a new strategy used by OV20.0 to evade antiviral responses via PKR.IMPORTANCE Viruses evolve specific strategies to counteract host innate immunity. ORFV, an important zoonotic pathogen, encodes OV20.0 to suppress PKR activation via multiple mechanisms, including interactions with PKR and two PKR activators. In this study, we demonstrated that OV20.0 interacts with ADAR1, a cellular enzyme responsible for converting adenosine (A) to inosine (I) in RNA. The RNA binding domains, but not the catalytic domain, of ADAR1 are required for this interaction. The OV20.0-ADAR1 association affects the functions of both proteins; OV20.0 suppressed the A-to-I editing of ADAR1, while ADAR1 elevated OV20.0 expression. The proviral role of ADAR1 is likely due to the inhibition of PKR phosphorylation. As RNA editing by ADAR1 contributes to the stability of the genetic code and the structure of RNA, these observations suggest that in addition to serving as a PKR inhibitor, OV20.0 might modulate ADAR1-dependent gene expression to combat antiviral responses or achieve efficient viral infection.


Functional characterization of Aspergillus nidulans ypkA, a homologue of the mammalian kinase SGK.
The serum- and glucocorticoid-regulated protein kinase (SGK) is an AGC kinase involved in signal cascades regulated by glucocorticoid hormones and serum in mammals. The Saccharomyces cerevisiae ypk1 and ypk2 genes were identified as SGK homologues and Ypk1 was shown to regulate the balance of sphingolipids between the inner and outer plasma membrane. This investigation characterized the Aspergillus nidulans YPK1 homologue, YpkA, representing the first filamentous fungal YPK1 homologue. Two conditional mutant strains were constructed by replacing the endogenous ypk1 promoter with two different regulatable promoters, alcA (from the alcohol dehydrogenase gene) and niiA (from the nitrate reductase gene). Both constructs confirmed that ypkA was an essential gene in A. nidulans. Repression of ypkA caused decreased radial growth, a delay in conidial germination, deficient polar axis establishment, intense branching during late stages of growth, a lack of asexual spores, and a terminal phenotype. Membrane lipid polarization, endocytosis, eisosomes and vacuolar distribution were also affected by ypkA repression, suggesting that YpkA plays a role in hyphal morphogenesis via coordinating the delivery of cell membrane and wall constituents to the hyphal apex. The A. nidulans Pkh1 homologue pkhA was also shown to be an essential gene, and preliminary genetic analysis suggested that the ypkA gene is not directly downstream of pkhA or epistatic to pkhA, rather, ypkA and pkhA are genetically independent or in parallel. BarA is a homologue of the yeast Lag1 acyl-CoA-dependent ceramide synthase, which catalyzes the condensation of phytosphingosine with a fatty acyl-CoA to form phytoceramide. When barA was absent, ypkA repression was lethal to the cell. Therefore, there appears to be a genetic interaction between ypkA, barA, and the sphingolipid synthesis. Transcriptional profiling of ypkA overexpression and down-regulation revealed several putative YpkA targets associated with the observed phenotypes.


An MRI-based method to register patient-specific wall shear stress data to histology.
Wall shear stress (WSS), the frictional force exerted on endothelial cells by blood flow, is hypothesised to influence atherosclerotic plaque growth and composition. We developed a methodology for image registration of MR and histology images of advanced human carotid plaques and corresponding WSS data, obtained by MRI and computational fluid dynamics. The image registration method requires four types of input images, in vivo MRI, ex vivo MRI, photographs of transversally sectioned plaque tissue and histology images. These images are transformed to a shared 3D image domain by applying a combination of rigid and non-rigid registration algorithms. Transformation matrices obtained from registration of these images are used to transform subject-specific WSS data to the shared 3D image domain as well. WSS values originating from the 3D WSS map are visualised in 2D on the corresponding lumen locations in the histological sections and divided into eight radial segments. In each radial segment, the correlation between WSS values and plaque composition based on histological parameters can be assessed. The registration method was successfully applied to two carotid endarterectomy specimens. The resulting matched contours from the imaging modalities had Hausdorff distances between 0.57 and 0.70 mm, which is in the order of magnitude of the in vivo MRI resolution. We simulated the effect of a mismatch in the rigid registration of imaging modalities on WSS results by relocating the WSS data with respect to the stack of histology images. A 0.6 mm relocation altered the mean WSS values projected on radial bins on average by 0.59 Pa, compared to the output of original registration. This mismatch of one image slice did not change the correlation between WSS and plaque thickness. In conclusion, we created a method to investigate correlations between WSS and plaque composition.


Authorship Order in Medical Education Publications: In Search of Practical Guidance for the Community.
Phenomenon: With scholarly collaborations come questions about the order of authorship. Authorship order is an important consideration because it often used as an indicator of seniority, expertise, leadership, and scholarly productivity. As a result, authorship order factors into decisions about hiring, salary, resource allocation, and professional advancement. This study describes principles commonly applied to authorship order decisions within the medical education community and educators' opinions about the significance of authorship order. Approach: A questionnaire was developed to ascertain current practices related to authorship decisions. Sixteen items were rated in terms of frequency of actual use and the desirability of the practice using a 4-point rating scale: 1 (never), 2 (sometimes), 3 (often), and 4 (always). Additional questions addressed the perceived significance of authorship order. The last set of questions provided information about respondents' personal and career characteristics. The survey was delivered via e-mail to a random sample of 391 subscribers from the DR-ED listserv. Findings: Fifty-four e-mail addresses were returned as undeliverable; of the remaining 337 mailed surveys, 109 responses (32.3%) were received. Five of the current practices for determining authorship order were rated as both frequent and desirable; 4 items had low ratings suggesting that these practices were both infrequent and undesirable. For 7 items, there was a significant gap between the ratings of practice frequency and desirability. When asked about preferred authorship order strategies, most respondents (94%) endorsed listing authors by descending order based on contribution but were split in identifying the last author (47%) or second author (46%) as the next best placement after first author. Respondents supported the practice of many journals requiring authors to disclose their contributions, agreeing (69%) that it provides useful information for promotion and tenure committees; however, 43% were uncertain about how disclosed contributions were reflected in authorship order. Insights: Respondents strongly endorsed the importance of listing authors in order of decreasing contribution, although the meaning of second versus last author lacks consensus. This finding, together the other strategies that received strong endorsement and those that were not strongly endorsed, provides a starting point to develop guidance for medical educators about how to determine authorship order. Clear guidance for authors would promote fairness and accountability within the medical education community as well as provide more consistent interpretation for those who consider authorship order for career- and resource-related decisions.


Microplastics: What happens in the human digestive tract? First evidences in adults using in vitro gut models.
Microplastics (MPs) are ubiquitous in the environment and humans are inevitably exposed to them. However, the effects of MPs in the human digestive environment are largely unknown. The aim of our study was to investigate the impact of repeated exposure to polyethylene (PE) MPs on the human gut microbiota and intestinal barrier using, under adult conditions, the Mucosal Artificial Colon (M-ARCOL) model, coupled with a co-culture of intestinal epithelial and mucus-secreting cells. The composition of the luminal and mucosal gut microbiota was determined by 16S metabarcoding and microbial activities were characterized by gas, short chain fatty acid, volatolomic and AhR activity analyses. Gut barrier integrity was assessed via intestinal permeability, inflammation and mucin synthesis. First, exposure to PE MPs induced donor-dependent effects. Second, an increase in abundances of potentially harmful pathobionts, Desulfovibrionaceae and Enterobacteriaceae, and a decrease in beneficial bacteria such as Christensenellaceae and Akkermansiaceae were observed. These bacterial shifts were associated with changes in volatile organic compounds profiles, notably characterized by increased indole 3-methyl- production. Finally, no significant impact of PE MPs mediated by changes in gut microbial metabolites was reported on the intestinal barrier. Given these adverse effects of repeated ingestion of PE MPs on the human gut microbiota, studying at-risk populations like infants would be a valuable advance.


[The repair processes and cell populations relationship in bone marrow of patients with acute radiation injury: morphological study].
The results of bone marrow pathomorphologic investigation in patients who died of acute radiation sickness (3.7-13.7 Gy) after the Chernobyl accident are presented. Stratification into three consecutive periods was performed according to the length of postexposure survival (14-96 days) and the morphologic picture dynamics: 1) bone marrow aplasia (2-3 weeks after exposure) with a domination of stromal and plasma cell populations, exhibiting the morphologic characteristics of functional activity; 2) initial repair period (3-4 weeks after exposure) was characterized by the beginning of hematopoietic proliferation with simultaneous occurrence of cell death via apoptosis; 3) active repair period (4 and more weeks after exposure) was defined by increased marrow cellularity (trilineage regeneration), accompanied by decreased number of plasma cells and occurrence of large granular lymphocytes.


Evaluation of decision rules to identify postmenopausal women for intervention related to osteoporosis.
Decision rules for intervention that utilize screening tools for bone mineral density (BMD) testing and incorporating the BMD findings and other risk factors to identify high-risk women to prevent fracture have not been evaluated. We examine the sensitivity and specificity of decision rules for intervention based on two pre-BMD screening tools: Simple Calculated Osteoporosis Risk Estimation (SCORE) and a Study of Osteoporotic Fractures (SOF)-based tool. Women 60 years of age and older without previous osteoporosis diagnosis were randomly selected from a managed care population and invited to receive a BMD test. Four hundred sixteen women had complete information and were included in the study. Women were classified as high risk requiring intervention using three different criteria: World Health Organization (t-score -2.5 or less), National Osteoporosis Foundation (t-score -2.0 or less, or -1.5 or less with one or more risk factors), and the SOF-based criteria (prior fracture; or age 60-64 with t-score less than -2.5 or age 65 or older with z-score less than -0.43 and five or more risk factors). SCORE identified 82% of the women as appropriate for BMD testing, whereas the SOF-based tool identified 26%. Sensitivity and specificity were 89.8%-96.5% and 23.8%-34.8%, respectively, for the decision rule using SCORE as the screening tool and 30.5%-84.9% and 76.0%-95.8%, respectively, for the decision rule based on SOF screening criteria. SCORE correctly identified more women who were at high risk for intervention, whereas the SOF-based tool correctly identified more women who do not meet intervention criteria. The appropriate selection of a screening tool depends upon the objective for intervention and trade-off between not identifying women for BMD testing who are at high risk and identifying more women for BMD testing who are at low risk.


The effect of stimulus duration on over-selectivity: Evidence for the role of within-compound associations.
The phenomenon whereby behavior becomes controlled by one aspect of the environment at the expense of other aspects of the environment (stimulus overselectivity) is widespread across many intellectual and developmental disabilities. However, the theoretical mechanisms underpinning overselectivity are not understood. Given similarities between overselectivity and overshadowing, exploring overselectivity using associative learning paradigms might allow better theoretical understanding of the phenomenon. Three experiments explored overselectivity using a simultaneous discrimination task with typically developing participants undergoing a cognitively demanding task. Experiment 1 investigated whether stimulus duration effects found within the overshadowing literature also occurred in an overselectivity paradigm, and demonstrated that greater overselectivity was observed when stimuli were presented for short durations (2s and 5s) compared with longer durations (10s). Experiment 2 demonstrated that a posttraining revaluation procedure resulted in retrospective revaluation for stimuli presented at shorter durations (2s) and mediated extinction for stimuli presented at longer durations (10s). Such results replicate findings from the overshadowing literature that have been interpreted in terms of within-compound associations while also supporting assumptions made by an extended comparator hypothesis. Experiment 3 uses an additional control condition to further demonstrate that the retrospective revaluation is a genuine revaluation effect. Additionally, the experiment provides further evidence for the within-compound association explanation of the results through manipulating the consistency with which elements of a compound were paired during training. Taken together, the findings highlight the necessity to consider the role of within-compound associations in overselectivity, allowing for a better understanding of overselectivity effects. (PsycINFO Database Record


Exploring the formation of gold/silver nanoalloys with gas-phase synthesis and machine-learning assisted simulations.
While nanoalloys are of paramount scientific and practical interest, the main processes leading to their formation are still poorly understood. Key structural features in the alloy systems, including the crystal phase, chemical ordering, and morphology, are challenging to control at the nanoscale, making it difficult to extend their use to industrial applications. In this contribution, we focus on the gold/silver system that has two of the most prevalent noble metals and combine experiments with simulations to uncover the formation mechanisms at the atomic level. Nanoparticles were produced using a state-of-the-art inert-gas aggregation source and analyzed using transmission electron microscopy and energy-dispersive X-ray spectroscopy. Machine-learning-assisted molecular dynamics simulations were employed to model the crystallization process from liquid droplets to nanocrystals. Our study finds a preponderance of nanoparticles with five-fold symmetric morphology, including icosahedra and decahedra which is consistent with previous results on mono-metallic nanoparticles. However, we observed that gold atoms, rather than silver atoms, segregate at the surface of the obtained nanoparticles for all the considered alloy compositions. These segregation tendencies are in contrast to previous studies and have consequences on the crystallization dynamics and the subsequent crystal ordering. We finally showed that the underpinning of this surprising segregation dynamics is due to charge transfer and electrostatic interactions rather than surface energy considerations.


Transcriptomic profiles of Japanese medaka (Oryzias latipes) in response to alkalinity stress.
Oryzias latipes (Adrianichthyidae), known as Japanese medaka or Japanese killifish, is a small 2-4 cm long fish common in rice paddies in coastal Southeast Asia and is also a popular aquarium fish. It has been widely used as a research model because of its small size and because it is very easy to rear. Alkalinity stress is considered to be one of the major stressors on fish in saline-alkaline water. As very little is known about molecular genetic responses of aquatic organisms to alkalinity stress, we examined genome-wide gene expression profiles of Japanese medaka in response to carbonate alkalinity stress. Adult fish were exposed to freshwater and high carbonate alkaline water in the laboratory. We designed a microarray containing 26,429 genes for measuring gene expression change in the gills of the fish exposed to high carbonate alkalinity stress. Among these genes, 512 were up-regulated and 501 were down-regulated in the gills. These differentially expressed genes can be divided into gene groups using gene ontology, including biological processes, cellular components and molecular function. These gene groups are related to acid-base and ion regulation, cellular stress response, metabolism, immune response, and reproduction processes. Biological pathways including amino sugar and nucleotide sugar metabolism, porphyrin and chlorophyll metabolism, metabolism of xenobiotics by cytochrome P450, drug metabolism, aminoacyl-tRNA biosynthesis, glycine, serine and threonine metabolism, ascorbate and aldarate metabolism, pentose and glucuronate interconversions, glutathione metabolism, and fructose and mannose metabolism were significantly up-regulated. Alkalinity stress stimulates the energy and ion regulation pathways, and it also slows down the pathways related to the immune system and reproduction.


Measuring Surface Pressure on Rotating Compressor Blades Using Pressure Sensitive Paint.
Pressure sensitive paint (PSP) was used to measure pressure on the blades of a radial compressor with a 51 mm inlet diameter rotating at speeds up to 50 krpm using the so called lifetime method. A diode laser with a scanning-mirror system was used to illuminate the paint and the luminescent lifetime was registered using a photo multiplier. With the described technique the surface-pressure fields were acquired for eight points in the compressor map, useful for general understanding of the flow field and for CFD validation. The PSP was of so called fast type, which makes it possible to observe pressure variations with frequencies up to several kHz. Through frequency spectrum analysis we were able to detect the pulsating flow frequency when the compressor was driven to surge.


Wintering greater scaup as biomonitors of metal contamination in federal wildlife refuges in the Long Island region.
Tissues of greater scaup (Aythya marila mariloides) and components of their habitat (sediment, plankton, macroalgae, and invertebrates) were collected for heavy metal analysis in the winter of 1996-97 from US Department of the Interior wildlife refuges in the Long Island region. Geographic and temporal relationships between the concentration of nine metals in tissue and in habitat components were examined. In greater scaup tissues and habitat components, concentrations of As and Se were highest in Branford, Connecticut; Pb values were greatest in Oyster Bay, New York; and Hg concentrations were largest in Sandy Hook, New Jersey. Over the course of the winter, the concentration of Hg in liver increased, and concentrations of Cd, Cr, Cu, Hg, Pb, Se, and Zn in kidney decreased. Based on several criteria derived from geographic and temporal trends, metals were ranked using the apparent biomonitoring efficacy of greater scaup (As = Cr > Cu = Pb = Zn = Hg > Se = Cd > Ni). Although the seasonal migration and daily mobility of greater scaup are drawbacks to using this species as a sentinel for metal pollution, it was possible to demonstrate a relationship between geographic and temporal patterns of metals in habitat and greater scaup tissue. However, most metal concentrations in tissue were below thresholds known to adversely affect health of waterfowl.


Low Dimensional Platinum-Based Bimetallic Nanostructures for Advanced Catalysis.
The development of renewable energy storage and conversion has been greatly promoted by the achievements in platinum (Pt)-based catalysts, which possess remarkable catalytic performance. However, the high cost and limited resources of Pt have hindered the practical applications and thus stimulated extensive efforts to achieve maximized catalytic performance with minimized Pt content. Low dimensional Pt-based bimetallic nanomaterials (such as nanoplates and nanowires) hold enormous potential to realize this target owing to their special atomic arrangement and electronic structures. Recent achievements reveal that strain engineering (e.g., the compressive or tensile strain existing on the Pt skin), surface engineering (e.g., high-index facets, Pt-rich surface, and highly open structures), and interface engineering (e.g., composition-segregated nanostructures) for such nanomaterials can readily lead to electronic modification, more active sites, and strong synergistic effect, thus opening up new avenues toward greatly enhanced catalytic performance. In this Account, we focus on recent advances in low dimensional Pt-based bimetallic nanomaterials as promising catalysts with high activity, long-term stability, and enhanced selectivity for both electrocatalysis and heterogeneous reactions. We begin by illustrating the important role of several strategies on optimizing the catalytic performance: (1) regulated electronic structure by strain effect, (2) increased active sites by surface modification, and (3) the optimized synergistic effect by interfacial engineering. First of all, a difference in atomic bonding strength can result in compressive or tensile force, leading to downshift or upshift of the d-band center. Such effects can be significantly amplified in low-dimensionally confined nanostructures, producing optimized bonding strength for improved catalysis. Furthermore, a high density of high-index facets and a Pt-rich surface in shape-controlled nanostructures based on surface engineering provide further enhancement due to the increased Pt atom utilization and optimal adsorption energy. Finally, interfacial engineering of low dimensional Pt-based bimetallic nanomaterials with high composition-segregation can facilitate the catalytic process due to a strong synergetic effect, which effectively tunes the electronic structure, modifies the coordination environment, and prevents catalysts from serious aggregation. The rational design of low dimensional Pt-based bimetallic nanomaterials with superior catalytic properties based on strain, surface, and interface engineering could help realize enhanced catalysis, gain deep understanding of the structure-performance relationship, and expand access to Pt-based materials for general communities of materials science, chemical engineering, and catalysis in renewable energy research fields.


In situ localization of CD83-positive dendritic cells in psoriatic lesions.
Dendritic cells (DC) are considered to be the most potent antigen-presenting cells, and CD83 is expressed at a high level on immune-competent, activated and mature DC. Although changes in the number or localization of mature and activated CD83+ DC could be expected in psoriasis, there is little information on such changes. Morphological identification of CD83+ DC in psoriatic skin lesions. Immunohistochemical staining was performed in 5 specimens of psoriasis vulgaris and 6 specimens of pustular psoriasis. Formalin-fixed, paraffin-embedded sections were used for examination in this study. The skin sections were pretreated with 0.1% trypsin for 60 min at 37 degrees C prior to immunostaining for CD83. A small but significant subpopulation of CD83+ DC was found in the upper dermis. In addition, CD83+ DC were occasionally scattered in the epidermis. The most common distribution pattern of CD83+ DC was as clusters with mononuclear lymphoid cells in the upper dermis. CD83+ DC were in close contact with lymphocytes. High-intensity staining of CD83 antigens was detected not only on the surface, but also in the cytoplasm of DC. These results indicate that activated and mature CD83+ DC may play a role in the immune response in psoriasis and provide in vivo support for the concept that CD83+ DC provide signals for direct intralesional T cell activation.


Respiratory chain complex I deficiency caused by mitochondrial DNA mutations.
Defects of the mitochondrial respiratory chain are associated with a diverse spectrum of clinical phenotypes, and may be caused by mutations in either the nuclear or the mitochondrial genome (mitochondrial DNA (mtDNA)). Isolated complex I deficiency is the most common enzyme defect in mitochondrial disorders, particularly in children in whom family history is often consistent with sporadic or autosomal recessive inheritance, implicating a nuclear genetic cause. In contrast, although a number of recurrent, pathogenic mtDNA mutations have been described, historically, these have been perceived as rare causes of paediatric complex I deficiency. We reviewed the clinical and genetic findings in a large cohort of 109 paediatric patients with isolated complex I deficiency from 101 families. Pathogenic mtDNA mutations were found in 29 of 101 probands (29%), 21 in MTND subunit genes and 8 in mtDNA tRNA genes. Nuclear gene defects were inferred in 38 of 101 (38%) probands based on cell hybrid studies, mtDNA sequencing or mutation analysis (nuclear gene mutations were identified in 22 probands). Leigh or Leigh-like disease was the most common clinical presentation in both mtDNA and nuclear genetic defects. The median age at onset was higher in mtDNA patients (12 months) than in patients with a nuclear gene defect (3 months). However, considerable overlap existed, with onset varying from 0 to >60 months in both groups. Our findings confirm that pathogenic mtDNA mutations are a significant cause of complex I deficiency in children. In the absence of parental consanguinity, we recommend whole mitochondrial genome sequencing as a key approach to elucidate the underlying molecular genetic abnormality.


Humans use continuous visual feedback from the hand to control both the direction and distance of pointing movements.
Vision of the hand during reaching provides dynamic feedback that can be used to control movement. We investigated the relative contributions of feedback about the direction and distance of the hand relative to a target. Subjects made pointing movements in a 3-D virtual environment, in which a small sphere provided dynamic visual feedback about the position of their unseen fingertip. On a subset of trials, the position of the virtual fingertip was smoothly shifted by 2 cm during movement, either (1) in the direction of movement, which would require adjustments to the distance moved, or (2) orthogonal to the direction of movement, which would require adjustments to the direction moved. Despite not noticing the perturbations, subjects adjusted their movements to compensate for both types of visual shifts. Corrective responses to direction perturbations were observed within 117 ms, and response latencies were invariant to movement speed and perturbation onset time. Initial corrections to distance perturbations were smaller and appeared after longer delays of 130-200 ms, and both the speed and magnitude of responses were reduced for early onset perturbations. Simulations of a feedback control model that optimally integrates visual information over time show that the results can be explained by differences in the sensory noise levels in the visual dimensions relevant for direction and distance control.


Monitoring for HHV-6 infection after renal transplantation: evaluation of risk factors for sustained viral replication.
Human herpesvirus-6 (HHV-6) is known to reactivate after renal transplantation and has been associated with several clinical manifestations. Risk factors for sustained viral replication, however, remain unclear. Thirty consecutive kidney transplant patients were prospectively followed for HHV-6 replication between February 2007 and February 2008. Plasma samples for DNA detection were collected from the donor and the recipient before transplantation and from the recipient weekly for the first 2 months after transplantation and then every 2 weeks for 2 additional months. HHV-6 active infection was defined as detection of viral DNA in plasma, by polymerase chain reaction, in at least two consecutive samples over an interval of at least 1 week. Active viral infection was detected in 25% of the recipients before transplantation and 27% (8 of 30) of the patients after transplantation. The mean time to onset of viral replication was 28.1 days after transplantation and 7 of 8 (87.5%) were asymptomatic. Risk factors associated with active HHV-6 infection were receiving an organ from a living donor (P=0.028), recipients with IgM antibodies detected before transplantation (P=0.005), and pretransplantation recipient HHV-6 viral load more than 10,000 copies/mL plasma (P=0.034). Active HHV-6 infection occurs early after renal transplantation and is mostly asymptomatic. Donor or recipient infection may occur at the time of transplantation and are related to higher rates of posttransplantation infections.


[The surgical treatment using inverted graft insertion technique for left ventricular aneurysm combined with aortic stenosis; report of a case].
An 83-year-old female with congestive heart failure[ New York Heart Association(NYHA) classification III ] due to left ventricular aneurysm, aortic stenosis( AS) with an aortic valve area of 0.66 cm2 and moderate tricuspid regurgitation, underwent endoventricular circular patch plasty using inverted graft insertion technique, aortic valve replacement (AVR) and tricuspid annuloplasty (TAP) successfully. It is highly probable that moderate aortic stenosis and localized apical transmural infarction had been gradually getting worse and they coincidentally caused both progressed AS and ventricular aneurismal change in our case.


Cell-Penetrating Dynamic-Covalent Benzopolysulfane Networks.
Cyclic oligochalcogenides (COCs) are emerging as promising systems to penetrate cells. Clearly better than and different to the reported diselenolanes and epidithiodiketopiperazines, we introduce the benzopolysulfanes (BPS), which show efficient delivery, insensitivity to inhibitors of endocytosis, and compatibility with substrates as large as proteins. This high activity coincides with high reactivity, selectively toward thiols, exceeding exchange rates of disulfides under tension. The result is a dynamic-covalent network of extreme sulfur species, including cyclic oligomers, from dimers to heptamers, with up to nineteen sulfurs in the ring. Selection from this unfolding adaptive network then yields the reactivities and selectivities needed to access new uptake pathways. Contrary to other COCs, BPS show high retention on thiol affinity columns. The identification of new modes of cell penetration is important because they promise new solutions to challenges in delivery and beyond.


Regulatory T cells in maintenance and reversal of peripheral tolerance in vivo.
Whether regulatory T cells could suppress the immune responses to an unrelated Ag and could induce tolerance to that Ag was investigated. In female B6 (I-Ab) mice tolerized to MHC class II I-Abm12 (bm12) alloantigen by cyclophosphamide-induced system, regulatory T cells specific for bm12 Ag were induced. These regulatory T cells suppressed in vivo immune responses to an unrelated Ag (male Ag) that was co-expressed with bm12 Ag on the same graft. However, the immune responses to male Ag suppressed by regulatory T cells were reverted by immunization with male Ag alone, indicating regulatory T cells suppressed the induction of immune responses to the unrelated Ag co-expressed with suppressogenic determinant, but did not inactivate T cells specific for that third Ag, such as clonal anergy or deletion. These results provided that minor Ag adjacent to major Ag, even if they were expressed on the professional APC, could be suppressed in the periphery, and the responses to those minor Ag could be activated when the suppression mechanism was abrogated or ineffective.


A preliminary in vitro evaluation of the bioactive potential of cryogel scaffolds incorporated with Manuka honey for the treatment of chronic bone infections.
Previous studies have identified honey as an agent in bacterial inhibition and a mediator in lowering the pH at the wound site. Manuka honey (MH), indigenous to New Zealand, contains a Unique Manuka Factor that provides an additional antibacterial agent. While there are many potential benefits to incorporating MH into wounds, there is currently no ideal way to deliver the material to the site of injury. Cryogels are a type of scaffold that possess high porosity, mechanical stability, and a sponge-like consistency. This study uniquely incorporates varying amounts of MH into cryogel scaffolds, utilizing its properties in a sustained release fashion to assist in the overall healing process, while using the cryogel structure as a tissue template. All cryogels were evaluated to determine the effects of MH on porosity, swelling potential, mechanical durability, and cell compatibility. The release of MH was also quantified to evaluate bacterial clearance potential, and the scaffolds were mineralized to replicate native bone. It was determined that a 5% MH silk fibroin cryogel has the potential to inhibit bacterial growth while still maintaining adequate porosity, mechanical properties, and cell infiltration. Such a scaffold would have use in a number of applications, including bone regeneration. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1918-1933, 2018.


Retinoic acid and tributyrin induce in-vitro radioiodine uptake and inhibition of cell proliferation in a poorly differentiated follicular thyroid carcinoma.
Radioiodine ablation is ineffective in patients with radioiodine nonresponsive thyroid carcinoma. We investigated the effects of all-trans retinoic acid (ATRA) combined with histone deacetylase inhibitor, tributyrin on sodium-iodide symporter (NIS) expression, radioiodine uptake, and inhibition of cell proliferation in a poorly differentiated follicular thyroid carcinoma (FTC-133) in vitro. FTC-133 cells were cultured in the presence of ATRA and tributyrin either as a single agent or in combinations for 48 h. The expression of NIS mRNA and protein was, respectively, detected by quantitative real-time polymerase chain reaction and western blot. The radioiodine uptake was determined after incubation of FTC-133 cells with 125I-iodide. Finally, the cell proliferation test of FTC-133 was performed after treatment. Enhanced expression of NIS mRNA and protein was observed in FTC-133 cells treated with ATRA and tributyrin, which further resulted in significant higher levels of radioiodine uptake than that of untreated control cells and cells treated with ATRA alone. Additive inhibition of the proliferation of FTC-133 cells was also observed with the combination of ATRA and tributyrin. The combination of ATRA and tributyrin induced a synergistic effect on radioiodine uptake and inhibition of FTC-133 cells proliferation in vitro. However, further in-vivo studies and additional molecular research will be needed to determine the absolute efficiency of radioiodine therapy.


Identification of vulnerable plaques and patients by intracoronary near-infrared spectroscopy and ultrasound (PROSPECT II): a prospective natural history study.
Near-infrared spectroscopy (NIRS) and intravascular ultrasound are promising imaging modalities to identify non-obstructive plaques likely to cause coronary-related events. We aimed to assess whether combined NIRS and intravascular ultrasound can identify high-risk plaques and patients that are at risk for future major adverse cardiac events (MACEs). PROSPECT II is an investigator-sponsored, multicentre, prospective natural history study done at 14 university hospitals and two community hospitals in Denmark, Norway, and Sweden. We recruited patients of any age with recent (within past 4 weeks) myocardial infarction. After treatment of all flow-limiting coronary lesions, three-vessel imaging was done with a combined NIRS and intravascular ultrasound catheter. Untreated lesions (also known as non-culprit lesions) were identified by intravascular ultrasound and their lipid content was assessed by NIRS. The primary outcome was the covariate-adjusted rate of MACEs (the composite of cardiac death, myocardial infarction, unstable angina, or progressive angina) arising from untreated non-culprit lesions during follow-up. The relations between plaques with high lipid content, large plaque burden, and small lumen areas and patient-level and lesion-level events were determined. This trial is registered with ClinicalTrials.gov, NCT02171065. Between June 10, 2014, and Dec 20, 2017, 3629 non-culprit lesions were characterised in 898 patients (153 [17%] women, 745 [83%] men; median age 63 [IQR 55-70] years). Median follow-up was 3·7 (IQR 3·0-4·4) years. Adverse events within 4 years occurred in 112 (13·2%, 95% CI 11·0-15·6) of 898 patients, with 66 (8·0%, 95% CI 6·2-10·0) arising from 78 untreated non-culprit lesions (mean baseline angiographic diameter stenosis 46·9% [SD 15·9]). Highly lipidic lesions (851 [24%] of 3500 lesions, present in 520 [59%] of 884 patients) were an independent predictor of patient-level non-culprit lesion-related MACEs (adjusted odds ratio 2·27, 95% CI 1·25-4·13) and non-culprit lesion-specific MACEs (7·83, 4·12-14·89). Large plaque burden (787 [22%] of 3629 lesions, present in 530 [59%] of 898 patients) was also an independent predictor of non-culprit lesion-related MACEs. Lesions with both large plaque burden by intravascular ultrasound and large lipid-rich cores by NIRS had a 4-year non-culprit lesion-related MACE rate of 7·0% (95% CI 4·0-10·0). Patients in whom one or more such lesions were identified had a 4-year non-culprit lesion-related MACE rate of 13·2% (95% CI 9·4-17·6). Combined NIRS and intravascular ultrasound detects angiographically non-obstructive lesions with a high lipid content and large plaque burden that are at increased risk for future adverse cardiac outcomes. Abbott Vascular, Infraredx, and The Medicines Company.


Detection of mosaic variants using genome sequencing in a large pediatric cohort.
The increased use of next-generation sequencing has expanded our understanding of the involvement and prevalence of mosaicism in genetic disorders. We describe a total of eleven cases: nine in which mosaic variants detected by genome sequencing (GS) and/or targeted gene panels (TGPs) were considered to be causative for the proband's phenotype, and two of apparent parental mosaicism. Variants were identified in the following genes: PHACTR1, SCN8A, KCNT1, CDKL5, NEXMIF, CUX1, TSC2, GABRB2, and SMARCB1. In addition, we identified one large duplication including three genes, UBE3A, GABRB3, and MAGEL2, and one large deletion including deletion of ARFGAP1, EEF1A2, CHRNA4, and KCNQ2. All patients were enrolled in the NYCKidSeq study, a research program studying the communication of genomic information in clinical care, as well as the clinical utility and diagnostic yield of GS for children with suspected genetic disorders in diverse populations in New York City. We observed variability in the correlation between reported variant allele fraction and the severity of the patient's phenotype, although we were not able to determine the mosaicism percentage in clinically relevant tissue(s). Although our study was not sufficiently powered to assess differences in mosaicism detection between the two testing modalities, we saw a trend toward better detection by GS as compared with TGP testing. This case series supports the importance of mosaicism in childhood-onset genetic conditions and informs guidelines for laboratory and clinical interpretation of mosaic variants detected by GS.


TORCH infections.
TORCH infections classically comprise toxoplasmosis, Treponema pallidum, rubella, cytomegalovirus, herpesvirus, hepatitis viruses, human immunodeficiency virus, and other infections, such as varicella, parvovirus B19, and enteroviruses. The epidemiology of these infections varies; in low-income and middle-income countries, TORCH infections are major contributors to prenatal, perinatal, and postnatal morbidity and mortality. Evidence of infection may be seen at birth, in infancy, or years later. For many of these pathogens, treatment or prevention strategies are available. Early recognition, including prenatal screening, is key. This article covers toxoplasmosis, parvovirus B19, syphilis, rubella, hepatitis B virus, hepatitis C virus, and human immunodeficiency virus.


CAMKII and calcineurin regulate the lifespan of Caenorhabditis elegans through the FOXO transcription factor DAF-16.
The insulin-like signaling pathway maintains a relatively short wild-type lifespan in Caenorhabditis elegans by phosphorylating and inactivating DAF-16, the ortholog of the FOXO transcription factors of mammalian cells. DAF-16 is phosphorylated by the AKT kinases, preventing its nuclear translocation. Calcineurin (PP2B phosphatase) also limits the lifespan of C. elegans, but the mechanism through which it does so is unknown. Herein, we show that TAX-6•CNB-1 and UNC-43, the C. elegans Calcineurin and Ca(2+)/calmodulin-dependent kinase type II (CAMKII) orthologs, respectively, also regulate lifespan through DAF-16. Moreover, UNC-43 regulates DAF-16 in response to various stress conditions, including starvation, heat or oxidative stress, and cooperatively contributes to lifespan regulation by insulin signaling. However, unlike insulin signaling, UNC-43 phosphorylates and activates DAF-16, thus promoting its nuclear localization. The phosphorylation of DAF-16 at S286 by UNC-43 is removed by TAX-6•CNB-1, leading to DAF-16 inactivation. Mammalian FOXO3 is also regulated by CAMKIIA and Calcineurin. DOI:http://dx.doi.org/10.7554/eLife.00518.001.


Mesodermal patterning activity of SCL.
The transcription factor SCL is critically required for establishing hematopoiesis and for proper endothelial development, but not for maintenance of hematopoietic stem cells or endothelial cells in the adult. Conflicting data exists regarding the developmental function of SCL, namely, whether it acts as a master regulator, actively patterning mesoderm toward hematopoietic development at the expense of other lineages, or is merely necessary to maintain the earliest committed hematopoietic precursors. To answer this question, we have engineered murine embryonic stem cells with a conditional doxycycline-inducible SCL transgene, and evaluated the effects of SCL expression at defined time points during in vitro development. Continual SCL expression during differentiation results in markedly increased hematopoiesis. By using pulses of gene expression over a 6-day differentiation time course, we map and characterize windows of SCL responsiveness. Notably, a pulse of SCL expression during early mesodermal patterning (48 to 72 hours of differentiation) promoted Flk1+ PDGFRalphaneg presumptive extraembryonic/lateral plate mesoderm at the expense of PDGFRalpha+ Flk1neg presumptive paraxial mesoderm. Consistent with this, the early pulse of SCL expression also expanded hematopoietic colony-forming cell numbers, while concomitantly repressing expression of paraxial and cardiac markers, and inhibited development of beating cardiomyocytes. By mixing the inducible embryonic stem cells with fluorescently labeled wild-type cells in chimeric embryoid bodies, we show that these effects of SCL are cell autonomous. These data support a master-regulatory role for SCL in mesodermal patterning, in addition to its established later roles in hematopoietic differentiation.


The human transmembrane secretory component (poly-Ig receptor): molecular cloning, restriction fragment length polymorphism and chromosomal sublocalization.
The human transmembrane secretory component (SC) mediates glandular translocation of polymeric IgA and IgM into exocrine secretions. A 2898-bp cDNA clone, encoding the entire sequence of the human transmembrane SC, was isolated from a colonic adenocarcinoma cell line cDNA library. The deduced amino-acid sequence had a length of 764 residues and showed an overall similarity of 56% and 64% with the rabbit and rat counterpart, respectively. A restriction fragment length polymorphism (RFLP) was found with PvuII, revealing a two-alle RFLP with an autosomal codominant inheritance pattern and allele frequencies of 0.65 and 0.35. Southern blot analysis of human-rodent somatic hybrid panels, including hybrids with translocation chromosomes carrying different parts of chromosome 1, assigned the SC gene to 1q31-q42, thus confirming a previously reported provisional assignment.


A proto-dictionary of psychoanalysis.
In this paper I take Freud's first published book, his monograph 'On aphasia', and look carefully at the terms he used in it, suggesting some later reappearances of the conception expressed by that particular word. There are several words that entered the analytic vocabulary with the monograph, indicating, as Stengel proposed, that the book 'laid the foundations of psychoanalysis'. I review the following words: association, divided attention, cathexis, complex, connection, physiological correlate, impulse to speak, memory-image, primary, representation, self-observation, spontaneous speech and transference.


Recent Surgical and Medical Advances in the Treatment of Dupuytren's Disease - A Systematic Review of the Literature.
Dupuytren's disease (DD) is a type of fibromatosis which progressively results in the shortening and thickening of the fibrous tissue of the palmar fascia. This condition which predominantly affects white-northern Europeans has been identified since 1614. DD can affect certain activities of daily living such as face washing, combing hair and putting hand in a glove. The origin of Dupuytren's contracture is still unknown, but there are a number of treatments that doctors have come across throughout the years. Historically surgery has been the mainstay treatment for DD but not the only one. The objective is to make a structured review of the most recent advances in treatment of DD including the surgical and medical interventions. We have looked at the most relevant published articles regarding the various treatment options for DD. This review has taken 55 articles into consideration which have met the inclusion criteria. The most recent treatments used are multi-needle aponeurotomy, extensive percutaneous aponeurotomy and lipografting, injecting collagenase Clostridium histolyticum, INF-gamma and shockwave therapy as well as radiotherapy. Each of these treatments has certain advantages and drawbacks and cannot be used for every patient. In order to prevent this condition, spending more time and money in the topic is required to reach better and more consistent treatments and ultimately to eradicate this disease.


Detection of H3+ in the diffuse interstellar medium toward Cygnus OB2 No. 12.
The molecular ion H3+ is considered the cornerstone of interstellar chemistry because it initiates the reactions responsible for the production of many larger molecules. Recently discovered in dense molecular clouds, H3+ has now been observed in the diffuse interstellar medium toward Cygnus OB2 No. 12. Analysis of H3+ chemistry suggests that the high H3+ column density (3.8 x 10(14) per square centimeter) is due not to a high H3+ concentration but to a long absorption path. This and other work demonstrate the ubiquity of H3+ and its potential as a probe of the physical and chemical conditions in the interstellar medium.


Contrary to Psychological and Popular Opinion, There Is No Compelling Evidence That Older Adults Are Disproportionately Victimized by Consumer Fraud.
According to psychological and popular opinion, older persons are especially susceptible to consumer fraud. Research on cognitive and affective aging reveals age-related changes that could increase the vulnerability of older persons to consumer fraud. However, this research does not show that consumer fraud actually is more prevalent among older persons. In generalizing from laboratory findings of cognitive decline to age differences in the prevalence of consumer fraud, psychologists may underestimate the influence in everyday life of possible protective factors associated with old age, including increased experience and changes in goals, lifestyle, income, as well as purchasing and risk behaviors. We review evidence on the prevalence of consumer fraud and conclude that there is no clear indication that it is more prevalent among older persons. Aggregating across all consumer frauds, there is evidence that consumer fraud is less common among older persons than adults of other ages. This evidence is potentially flawed, however, because of failings inherent in the methodologies. In the absence of irrefutable data, it is premature to conclude that consumer fraud is less prevalent among older adults, but it is also premature to conclude that consumer fraud is more prevalent among older persons, as is assumed in conventional and psychological wisdom.


Hemoglobin Raleigh as the cause of a falsely increased hemoglobin A1C in an automated ion-exchange HPLC method.
Irreversible glycation of the hemoglobin A0 (HbA0) beta chain leads to the production of HbA1C, which can be used to monitor long-term blood glucose control in patients with diabetes mellitus. HbA1C is less positively charged than nonglycated HbA0, and this decrease in charge is the basis of ion-exchange and electrophoretic methods that measure HbA1C. We recently identified a sample that appeared to contain 46% HbA1C by an automated ion-exchange HPLC method (Bio-Rad Variant) but only 3.8% by an immunoinhibition latex agglutination method. A combination of traditional and mass spectrometric protein analysis and genomic DNA analysis of the Hb beta chain and genes revealed that the patient was heterozygotic for Hb-Raleigh, a variant containing a valine-->alanine substitution at position 1 of the beta chain. The amino-terminal alanine in this variant Hb is posttranslationally modified by acetylation, leading to a charge difference similar to glycation and making the behavior of HbA1C and Hb Raleigh virtually identical in the ion-exchange HPLC method. This observation suggests that it is important to confirm HbA1C values in excess of 15%, especially if they are not consistent with the clinical picture, by an independent HbA1C method such as immunoassay or boronic acid affinity chromatography. However, for this particular variant Hb, even these latter methods might be misleading, because the acetylated N-terminal amino acid of the Hb-Raleigh beta chain cannot be glycated.


Experimental Constraint on an Exotic Spin- and Velocity-Dependent Interaction in the Sub-meV Range of Axion Mass with a Spin-Exchange Relaxation-Free Magnetometer.
We conducted a search for an exotic spin- and velocity-dependent interaction for polarized electrons with an experimental approach based on a high-sensitivity spin-exchange relaxation-free (SERF) magnetometer, which serves as both a source of polarized electrons and a magnetic-field sensor. The experiment aims to sensitively detect magnetic-fieldlike effects from the exotic interaction between the polarized electrons in a SERF vapor cell and unpolarized nucleons of a closely located solid-state mass. We report experimental results on the interaction with 82 h of data averaging, which sets an experimental limit on the coupling strength around 10^{-19} for the axion mass m_{a}≲10^{-3} eV, within the important axion window.


Type 2 iodothyronine deiodinase is upregulated in rat slow- and fast-twitch skeletal muscle during cold exposure.
During cold acclimation, shivering is progressively replaced by nonshivering thermogenesis. Brown adipose tissue (BAT) and skeletal muscle are relevant for nonshivering thermogenesis, which depends largely on thyroid hormone. Since the skeletal muscle fibers progressively adapt to cold exposure through poorly defined mechanisms, our intent was to determine whether skeletal muscle type 2 deiodinase (D2) induction could be implicated in the long-term skeletal muscle cold acclimation. We demonstrate that in the red oxidative soleus muscle, D2 activity increased 2.3-fold after 3 days at 4°C together with the brown adipose tissue D2 activity, which increased 10-fold. Soleus muscle and BAT D2 activities returned to the control levels after 10 days of cold exposure, when an increase of 2.8-fold in D2 activity was detected in white glycolytic gastrocnemius but not in red oxidative gastrocnemius fibers. Propranolol did not prevent muscle D2 induction, but it impaired the decrease of D2 in BAT and soleus after 10 days at 4°C. Cold exposure is accompanied by increased oxygen consumption, UCP3, and PGC-1α genes expression in skeletal muscles, which were partialy prevented by propranolol in soleus and gastrocnemius. Serum total and free T3 is increased during cold exposure in rats, even after 10 days, when BAT D2 is already normalized, suggesting that skeletal muscle D2 activity contributes significantly to circulating T3 under this adaptive condition. In conclusion, cold exposure is accompanied by concerted changes in the metabolism of BAT and oxidative and glycolytic skeletal muscles that are paralleled by type 2 deiodinase activation.


Niacin restriction with NAMPT-inhibition is synthetic lethal to neuroendocrine carcinoma.
Nicotinamide phosphoribosyltransferase (NAMPT) plays a major role in NAD biosynthesis in many cancers and is an attractive potential cancer target. However, factors dictating therapeutic efficacy of NAMPT inhibitors (NAMPTi) are unclear. We report that neuroendocrine phenotypes predict lung and prostate carcinoma vulnerability to NAMPTi, and that NAMPTi therapy against those cancers is enhanced by dietary modification. Neuroendocrine differentiation of tumor cells is associated with down-regulation of genes relevant to quinolinate phosphoribosyltransferase-dependent de novo NAD synthesis, promoting NAMPTi susceptibility in vitro. We also report that circulating nicotinic acid riboside (NAR), a non-canonical niacin absent in culture media, antagonizes NAMPTi efficacy as it fuels NAMPT-independent but nicotinamide riboside kinase 1-dependent NAD synthesis in tumors. In mouse transplantation models, depleting blood NAR by nutritional or genetic manipulations is synthetic lethal to tumors when combined with NAMPTi. Our findings provide a rationale for simultaneous targeting of NAR metabolism and NAMPT therapeutically in neuroendocrine carcinoma.


Voltage-gated proton channels.
Voltage-gated proton channels, HV1, have vaulted from the realm of the esoteric into the forefront of a central question facing ion channel biophysicists, namely, the mechanism by which voltage-dependent gating occurs. This transformation is the result of several factors. Identification of the gene in 2006 revealed that proton channels are homologues of the voltage-sensing domain of most other voltage-gated ion channels. Unique, or at least eccentric, properties of proton channels include dimeric architecture with dual conduction pathways, perfect proton selectivity, a single-channel conductance approximately 10(3) times smaller than most ion channels, voltage-dependent gating that is strongly modulated by the pH gradient, ΔpH, and potent inhibition by Zn(2+) (in many species) but an absence of other potent inhibitors. The recent identification of HV1 in three unicellular marine plankton species has dramatically expanded the phylogenetic family tree. Interest in proton channels in their own right has increased as important physiological roles have been identified in many cells. Proton channels trigger the bioluminescent flash of dinoflagellates, facilitate calcification by coccolithophores, regulate pH-dependent processes in eggs and sperm during fertilization, secrete acid to control the pH of airway fluids, facilitate histamine secretion by basophils, and play a signaling role in facilitating B-cell receptor mediated responses in B-lymphocytes. The most elaborate and best-established functions occur in phagocytes, where proton channels optimize the activity of NADPH oxidase, an important producer of reactive oxygen species. Proton efflux mediated by HV1 balances the charge translocated across the membrane by electrons through NADPH oxidase, minimizes changes in cytoplasmic and phagosomal pH, limits osmotic swelling of the phagosome, and provides substrate H(+) for the production of H2O2 and HOCl, reactive oxygen species crucial to killing pathogens.


Maternal choline supplementation protects against age-associated cholinergic and GABAergic basal forebrain neuron degeneration in the Ts65Dn mouse model of Down syndrome and Alzheimer's disease.
Down syndrome (DS) is a genetic disorder caused by triplication of human chromosome 21. In addition to intellectual disability, DS is defined by a premature aging phenotype and Alzheimer's disease (AD) neuropathology, including septohippocampal circuit vulnerability and degeneration of basal forebrain cholinergic neurons (BFCNs). The Ts65Dn mouse model recapitulates key aspects of DS/AD pathology, namely age-associated atrophy of BFCNs and cognitive decline in septohippocampal-dependent behavioral tasks. We investigated whether maternal choline supplementation (MCS), a well-tolerated treatment modality, protects vulnerable BFCNs from age- and genotype-associated degeneration in trisomic offspring. We also examined the effect of trisomy, and MCS, on GABAergic basal forebrain parvalbumin neurons (BFPNs), an unexplored neuronal population in this DS model. Unbiased stereological analyses of choline acetyltransferase (ChAT)-immunoreactive BFCNs and parvalbumin-immunoreactive BFPNs were conducted using confocal z-stacks of the medial septal nucleus and the vertical limb of the diagonal band (MSN/VDB) in Ts65Dn mice and disomic (2N) littermates at 3-4 and 10-12 months of age. MCS trisomic offspring displayed significant increases in ChAT-immunoreactive neuron number and density compared to unsupplemented counterparts, as well as increases in the area of the MSN/VDB occupied by ChAT-immunoreactive neuropil. MCS also rescued BFPN number and density in Ts65Dn offspring, a novel rescue of a non-cholinergic cell population. Furthermore, MCS prevented age-associated loss of BFCNs and MSN/VDB regional area in 2N offspring, indicating genotype-independent neuroprotective benefits. These findings demonstrate MCS provides neuroprotection of vulnerable BFCNs and non-cholinergic septohippocampal BFPNs, indicating this modality has translational value as an early life therapy for DS, as well as extending benefits to the aging population at large.


Five-year efficacy and safety of asfotase alfa therapy for adults and adolescents with hypophosphatasia.
Hypophosphatasia (HPP) features low tissue-nonspecific alkaline phosphatase (TNSALP) isoenzyme activity resulting in extracellular accumulation of its substrates including pyridoxal 5'-phosphate (PLP), the principal circulating form of vitamin B6, and inorganic pyrophosphate (PPi), a potent inhibitor of mineralization. Asfotase alfa is an enzyme replacement therapy developed to treat HPP. This multinational, randomized, open-label study (NCT01163149; EudraCT 2010-019850-42) evaluated the efficacy and safety of asfotase alfa in adults and adolescents 13-66 years of age with HPP. The study comprised a 6-month primary treatment period and a 4.5-year extension phase. In the primary treatment period, 19 patients were randomized to receive asfotase alfa 0.3 mg/kg/d subcutaneously (SC; n = 7), asfotase alfa 0.5 mg/kg/d SC (n = 6), or no treatment (control; n = 6) for 6 months. In the extension phase, patients received asfotase alfa (0.5 mg/kg/d for 6 mo-1 y, then 1 mg/kg/d 6 d/wk). During the primary treatment period, changes from Baseline to Month 6 in plasma PLP and PPi concentrations (coprimary efficacy measure) were greater in the combined asfotase alfa group compared with the control group, reaching statistical significance for PLP (P = 0.0285) but not for PPi (P = 0.0715). However, for the total cohort, the within subject changes in both PLP and PPi after 6 months and over 5 years of treatment with asfotase alfa were significant (P < 0.05). Secondary efficacy measures included transiliac crest histomorphometry, dual-energy X-ray absorptiometry (DXA), and the 6-Minute Walk Test (6MWT). A significant decrease from Baseline in mineralization lag time was observed in the combined asfotase alfa group at Year 1. There were no significant differences between treated and control patients in DXA mean bone mineral density results at 6 months; Z-scores and T-scores were within the expected range for age at Baseline and remained so over 5 years of treatment. On the 6MWT, median (min, max) distance walked increased from 355 (10, 620; n = 19) meters before treatment to 450 (280, 707; n = 13) meters at 5 years (P < 0.05). Results for the exploratory outcome measures suggested improvements in gross motor function, muscle strength, and patient-reported functional disability over 5 years of treatment. There were no deaths during this study. Asfotase alfa was generally well tolerated; the most common adverse events were mild to moderate injection site reactions. This study suggests that in adults and adolescents with pediatric-onset HPP, treatment with asfotase alfa is associated with normalization of circulating TNSALP substrate levels and improved functional abilities.


Predatory activity of the fungi Duddingtonia flagrans, Monacrosporium thaumasium, Monacrosporium sinense and Arthrobotrys robusta on Angiostrongylus vasorum first-stage larvae.
Angiostrongylus vasorum is a nematode that parasitizes domestic dogs and wild canids. We compared the predatory capacity of isolates from the predatory fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on first-stage larvae (L1) of A. vasorum under laboratory conditions. L1 A. vasorum were plated on 2% water-agar (WA) Petri dishes marked into 4 mm diameter fields with the four grown isolates and a control without fungus. Plates of treated groups contained each 1000 L1 A. vasorum and 1000 conidia of the fungal isolates AC001, NF34, SF53 and I31 on 2% WA. Plates of the control group (without fungus) contained only 1000 L1 A. vasorum on 2% WA. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for 7 days. Nematophagous fungi were not observed in the control group during the experiment. There was no variation in the predatory capacity among the tested fungal isolates (P>0.05) during the 7 days of the experiment. There was a significant reduction (P < 0.05) of 80.3%, 74.5%, 74.2% and 71.8% in the means of A. vasorum L1 recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively, compared to the control without fungi. In this study, the four isolates of predatory fungi were efficient in the in vitro capture and destruction of A. vasorum L1, confirming previous work on the efficiency of nematophagous fungi in the control of nematode parasites of dogs and as a possible alternative method of biological control.


Movements of the pelvis and lumbar spine during walking in people with acute low back pain.
Little is known about how acute low back pain affects pelvic and lumbar movements during walking. The aim of the present study was to determine if measurement of the amplitude of the angular movements of the pelvis and lumbar spine during walking is useful in the evaluation of people with acute low back pain. The study used a repeated-measures and correlational design; 11 individuals with low back pain (tested in the acute phase and six weeks later when symptoms had resolved) and matched control subjects were tested during treadmill walking. A video-analysis system was used to measure the amplitudes of movements of the pelvis and lumbar spine during walking. Pain level was measured with a visual analogue scale (VAS). During walking, movements of the pelvis (axial rotation) and lumbar spine (lateral flexion) were reduced in people with acute back pain compared with the resolved condition, but were not different from a group without a history of back pain. The amplitudes of the frontal plane movements of the pelvis and lumbar spine were negatively correlated with the intensity of pain (r(s) = -0.74). Measurement of pelvic and lumbar movements during walking is unlikely to have useful clinical applications for individuals, or when discriminating between impaired and unimpaired people, but may be applied to groups for hypothesis testing in evaluating change in back pain over time. An hypothesized model to explain the observed movements has been proposed.


Durable molecular response to imatinib mesylate following nonmyeloablative allogeneic stem-cell transplantation for persisting myeloid blast crisis in chronic myeloid leukemia.
We report a chronic myeloid leukemia (CML) patient in chronic phase (CP) who developed blast crisis (BC) under imatinib mesylate administered in a dose reduced and non-continuous fashion because of hematologic intolerance. The patient underwent nonmyeloablative stem-cell transplant from a matched unrelated donor, but failed to achieve full donor chimerism and antileukemic response resulting in persistence of advanced disease. Complete hematologic, cytogenetic and molecular responses were attained 5 weeks after readministration of regularly dosed imatinib and two-step nested RT-PCR confirmed molecular remission throughout a 6 month follow-up period. This is the first case demonstrating that imatinib mesylate is a highly effective and safe treatment option to induce durable molecular remission in patients with CML who remain in myeloid blast crisis after nonmyeloablative allogeneic stem-cell transplantation.


Biosorption of nanoparticles to heterotrophic wastewater biomass.
Sorption to activated sludge is a major removal mechanism for pollutants, including manufactured nanoparticles (NPs), in conventional activated sludge wastewater treatment plants. The objectives of this work were to (1) image sorption of fluorescent NPs to wastewater biomass; (2) quantify and compare biosorption of different types of NPs exposed to wastewater biomass; (3) quantify the effects of natural organic matter (NOM), extracellular polymeric substances (EPS), surfactants, and salt on NP biosorption; and (4) explore how different surface functionalities for fullerenes affect biosorption. Batch sorption isotherm experiments were conducted with activated sludge as sorbent and a total of eight types of NPs as sorbates. Epifluorescence images clearly show the biosorption of fluorescent silica NPs; the greater the concentration of NPs exposed to biomass, the greater the quantity of NPs that biosorb. Furthermore, biosorption removes different types of NPs from water to different extents. Upon exposure to 400 mg/L total suspended solids (TSS) of wastewater biomass, 97% of silver nanoparticles were removed, probably in part by aggregation and sedimentation, whereas biosorption was predominantly responsible for the removal of 88% of aqueous fullerenes, 39% of functionalized silver NPs, 23% of nanoscale titanium dioxide, and 13% of fullerol NPs. Of the NP types investigated, only aq-nC(60) showed a change in the degree of removal when the NP suspension was equilibrated with NOM or when EPS was extracted from the biomass. Further study of carbonaceous NPs showed that different surface functionalities affect biosorption. Thus, the production and transformations in NP surface properties will be key factors in determining their fate in the environment.


Clinical, virological and epidemiological characterization of dengue outbreak in Myanmar, 2015.
Hospital-based surveillance was conducted at two widely separated regions in Myanmar during the 2015 dengue epidemic. Acute phase serum samples were collected from 332 clinically diagnosed dengue patients during the peak season of dengue cases. Viremia levels were measured by quantitative real-time PCR and plaque assays using FcγRIIA-expressing and non-FcγRIIA-expressing BHK cells to specifically determine the infectious virus particles. By serology and molecular techniques, 280/332 (84·3%) were confirmed as dengue patients. All four serotypes of dengue virus (DENV) were isolated from among 104 laboratory-confirmed patients including two cases infected with two DENV serotypes. High percentage of primary infection was noted among the severe dengue patients. Patients with primary infection or DENV IgM negative demonstrated significantly higher viral loads but there was no significant difference among the severity groups. Viremia levels among dengue patients were notably high for a long period which was assumed to support the spread of the virus by the mosquito vector during epidemic. Phylogenetic analyses of the envelope gene of the epidemic strains revealed close similarity with the strains previously isolated in Myanmar and neighboring countries. DENV-1 dominated the epidemic in 2015 and the serotype (except DENV-3) and genotype distributions were similar in both study sites.


The changes in early phase insulin secretion in newly diagnosed, drug naive korean prediabetes subjects.
There have been no systematic observations regarding changes in early phase insulin secretion among Korean prediabetes and early stage type 2 diabetes mellitus (T2DM) patients. We conducted 75-g oral glucose tolerance tests (OGTT) in 873 subjects with suspected abnormal glucose tolerance. All subjects were diagnosed as having normal glucose tolerance (NGT), prediabetes (preDM), or T2DM according to the OGTT results and the insulin secretory and insulin resistance indices of each subject were calculated. Additionally, we analyzed the changes in early phase insulin secretion according to changes in fasting (Glc(0)), post-prandial (Glc(120)) glucose and HbA1c (A1c) levels. As compared to subjects with NGT, the insulin secretory indices of the preDM and T2DM subjects progressively declined, and the insulin resistance indices were progressively aggravated. Early phase insulin secretion decreased rapidly according to the increments of Glc(0), Glc(120) and A1c, and these changes were most prominent in the NGT stage. Compared to the control group, the early phase insulin secretion levels of the preDM or T2DM subjects were less than 50% when Glc(0) was over 100 mg/dL, Glc(120) was over 145 mg/dL, and A1c was over 5.8%. This study suggests that progressive beta cell dysfunction in Koreans may be initiated and rapidly aggravated during the period generally designated as 'normal.'


Different composition and mitotic activity of the haemopoietic tissue in bone marrow, spleen and liver in chronic myeloid leukaemia.
Aspirates from bone marrow, spleen and liver were analyzed in 15 untreated patients with chronic myeloid leukaemia (CML). The proportion of erythroblasts was higher in the spleen and liver than in the bone marrow. An increased frequency of basophilic leucocytes was recorded in the extramedullary sites compared to the bone marrow. In the liver aspirates a considerable admixture of lymphocytes was found. The mitotic indices of the granulopoietic and erythropoietic precursor cells were lower in the spleen and liver than in the bone marrow. The results indicate that the composition and proliferative activity of the haemopoietic tissue differ in bone marrow and extramedullary sites in CML. The discrepancies may be due to differences in the microenvironment for the haemopoietic cells or to a selective accumulation of cells with a low mitotic activity in extramedullary sites. Since the final blastic transformation of CML may originate outside the bone marrow, further studies of the extramedullary haemopoietic tissue may be worthwhile.


Role of cytokines in neurological disorders.
The balance between cytokines with pro- and anti-inflammatory effects contributes to the course of the Guillain-Barré syndrome and chronic inflammatory demyelinating polyneuropathy. TNFalpha seems to be an important factor in the cascade of events leading to demyelination and even axonal damage. During the acute phase, the serum concentrations of TNFalpha and IL-6 are elevated while anti-inflammatory cytokines are up-regulated in the recovery phase. Cytokines also have a key role in the pathogenesis of multiple sclerosis and most data suggest that this effect is mediated by myelin-specific CD4 T lymphocytes secreting Th type 1 cytokines. However, several different immune cells including B lymphocytes, CD8 T lymphocytes and NK T lymphocytes are also involved in the pathogenesis. Both Th1 and Th2 lymphocytes and cytokines probably participate in the development of myasthenia gravis (MG). The IFNalpha production is probably related to the severity of the disease, with clinical improvement associated with decreased production. The serum levels of IL-18 are significantly elevated in MG, with highest concentrations in patients with generalized disease. The immune system may be involved in the pathogenesis of AD by the effect of microglia, which can induce microglial activation with subsequent release of pro-inflammatory cytokines. In parkinsonism, there is evidence of chronic inflammation in the substantia nigra and striatum. Activated microglia, producing proinflammatory cytokines, surround the degenerating dopaminergic neurons and may contribute to the dopaminergic neuron loss. Studies of patients with epilepsy and animals with experimentally induced seizures indicate that cytokines may also influence the electrophysiological properties of neurons.


The meaning of nursing home friendships.
This research report on the social networks of institutionalized elderly people describes four types of networks and focuses on resident-resident interaction. Friendship themes of intimacy, social support, reciprocity, and companionship provide a basis for examining similarities in and differences between the network types. Discussion draws on the study results through examination of the meaning of friendship and interpretation of life-span themes.


X chromosome inactivation during induced differentiation of a female mouse embryonal carcinoma cell line.
P10 is a line of embryonal carcinoma cells with a euploid female karyotype. By making use of variant alleles of X-linked genes and of cytogenetic procedures, we have previously shown that the P10 cells have two genetically active X chromosomes. We show here that P10 cells rapidly differentiate into a cell type resembling extraembryonic endoderm when cultured in the presence of retinoic acid. This differentiation is accompanied by X chromosome inactivation as determined by the appearance of a late-replicating X chromosome. Analysis of the X-linked variant alleles indicated that the P10-derived endoderm did not preferentially inactivate paternally derived X chromosomes. This result is in contrast to the situation in normal extraembryonic endoderm, and suggests that the X inactivation process in differentiating P10 cultures resembles that which occurs in normal embryonic rather than extraembryonic tissues.


k-nearest neighbors directed noise injection in multilayer perceptron training.
The relation between classifier complexity and learning set size is very important in discriminant analysis. One of the ways to overcome the complexity control problem is to add noise to the training objects, increasing in this way the size of the training set. Both the amount and the directions of noise injection are important factors which determine the effectiveness for classifier training. In this paper the effect is studied of the injection of Gaussian spherical noise and -nearest neighbors directed noise on the performance of multilayer perceptrons. As it is impossible to provide an analytical investigation for multilayer perceptrons, a theoretical analysis is made for statistical classifiers. The goal is to get a better understanding of the effect of noise injection on the accuracy of sample-based classifiers. By both empirical as well as theoretical studies, it is shown that the -nearest neighbors directed noise injection is preferable over the Gaussian spherical noise injection for data with low intrinsic dimensionality.


Current guidelines for the management of non-alcoholic fatty liver disease: A systematic review with comparative analysis.
The current epidemic of non-alcoholic fatty liver disease (NAFLD) is reshaping the field of hepatology all around the world. The widespread diffusion of metabolic risk factors such as obesity, type2-diabetes mellitus, and dyslipidemia has led to a worldwide diffusion of NAFLD. In parallel to the increased availability of effective anti-viral agents, NAFLD is rapidly becoming the most common cause of chronic liver disease in Western Countries, and a similar trend is expected in Eastern Countries in the next years. This epidemic and its consequences have prompted experts from all over the word in identifying effective strategies for the diagnosis, management, and treatment of NAFLD. Different scientific societies from Europe, America, and Asia-Pacific regions have proposed guidelines based on the most recent evidence about NAFLD. These guidelines are consistent with the key elements in the management of NAFLD, but still, show significant difference about some critical points. We reviewed the current literature in English language to identify the most recent scientific guidelines about NAFLD with the aim to find and critically analyse the main differences. We distinguished guidelines from 5 different scientific societies whose reputation is worldwide recognised and who are representative of the clinical practice in different geographical regions. Differences were noted in: the definition of NAFLD, the opportunity of NAFLD screening in high-risk patients, the non-invasive test proposed for the diagnosis of NAFLD and the identification of NAFLD patients with advanced fibrosis, in the follow-up protocols and, finally, in the treatment strategy (especially in the proposed pharmacological management). These difference have been discussed in the light of the possible evolution of the scenario of NAFLD in the next years.


Acute myocardial infarction in the puerperium.
The authors describe the case of a forty-two-year-old patient with acute myocardial infarction (MI) on the third day of puerperium, who underwent direct angioplasty. The authors review the main physiological cardiovascular and hemostatic changes in pregnancy, which are usually well tolerated but which increase the probability of cardiovascular events when risk factors or previous cardiac disease are associated. Blood volume and cardiac output increase by around 50% during pregnancy, hemoglobin concentration falls, and reduced peripheral vascular resistance leads to a fall in arterial blood pressure during the first half of pregnancy. End-diastolic pressure, stroke volume and cardiac output increase in the immediate puerperium, and cardiovascular risk is highest in the perinatal period. The incidence of MI during pregnancy and puerperium is low; however, it is on the increase due to the growing number of pregnancies in women close to the end of their reproductive age.


Separation of protein charge variants by ultrafiltration.
The removal of product variants that form during downstream processing remains a challenge in the purification of recombinant therapeutic proteins. We examined the feasibility of separating variants with slightly different net charge using high-performance membrane ultrafiltration. A myoglobin variant was formed by reaction of the lysine epsilon-amino group with succinic anhydride. Sieving data were obtained over a range of solution conditions using commercial polyethersulfone ultrafiltration membranes. Maximum selectivity of about 7-fold was obtained at very low conductivity due to the strong electrostatic repulsion of the more negatively charged variant. Protein separations were performed by diafiltration. A two-stage process generated solutions of the normal myoglobin (in the permeate) and the charge variant (in the retentate), both at greater than 9-fold purification and 90% yield. These results provide the first demonstration that membrane systems can be used to separate proteins that differ by only a single charged amino acid residue.


Prevalence and pattern of renal bone disease in end stage renal disease patients in Ile-Ife, Nigeria.
Information on renal bone disease (RBD) is sparse in Nigeria. The prevalence of RBD in a dialysis population worldwide ranges between 33% and 67% and it increases with progression of renal insufficiency. To determine the prevalence and magnitude of RBD in patients with end stage renal disease (ESRD). Thirty consecutive ESRD patients were recruited. They had thorough clinical evaluation and investigations which included serum calcium, phosphate, alkaline phosphatase, albumin and skeletal survey. The serum iPTH, osteocalcin, and 1,25 (OH2) D3 were assessed in 20 patients. The patients were aged 18-72 years with a mean of 38.93+/-15.7 years. There was a male:female ratio of 4:1. Uraemic symptoms were the major presenting complaints. None of the patients complained of bone pain or fracture. The mean values for serum creatinine, urea, creatinine clearance, calcium, phosphate, albumin, alkaline phosphatase, iPTH, osteocalcin and 1,25 (OH)2 Vit D3 were 1478.96 +/- 771.12 micromol/L, 22.33 +/- 7.42 mmol/L, 3.38 +/-2.22 mls/min, 1.8 +/- 0.5 mmol/L, 1.61 +/- 0.65 mmol/L, 30.2 +/- 6.1 g/L, 124.33 +/- 63.37 IU/L, 22.66 +/- 24.72, 45.14 +/- 43.8, 37.7 +/- 22.3 respectively. There were hypocalcaemia and hyper-phosphataemia in 80% and 60% of the patients respectively. Alkaline phosphatase was elevated in 44% of the patients while 11.8% had hyperparathyroidism. Level of 1,25 (OH)2 Vit D3 was low in 83.3% of the patients. There was a significant negative correlation between serum calcium and iPTH levels (r = -0.915, p=0.029). There was also significant negative correlation between alkaline phosphatase and 1,25 (OH)2 Vit D3 and serum albumin. Radiological evidence of RBD occurred in only 16.7% of the patients. Renal bone disease is common in our patients with ESRD associated most commonly with low bone turnover while occurrence of hyperparathyroid bone disease appears low.


Opioid epidemic in the United States.
Over the past two decades, as the prevalence of chronic pain and health care costs have exploded, an opioid epidemic with adverse consequences has escalated. Efforts to increase opioid use and a campaign touting the alleged undertreatment of pain continue to be significant factors in the escalation. Many arguments in favor of opioids are based solely on traditions, expert opinion, practical experience and uncontrolled anecdotal observations. Over the past 20 years, the liberalization of laws governing the prescribing of opioids for the treatment of chronic non-cancer pain by the state medical boards has led to dramatic increases in opioid use. This has evolved into the present stage, with the introduction of new pain management standards by the Joint Commission on the Accreditation of Healthcare Organizations (JCAHO) in 2000, an increased awareness of the right to pain relief, the support of various organizations supporting the use of opioids in large doses, and finally, aggressive marketing by the pharmaceutical industry. These positions are based on unsound science and blatant misinformation, and accompanied by the dangerous assumptions that opioids are highly effective and safe, and devoid of adverse events when prescribed by physicians. Results of the 2010 National Survey on Drug Use and Health (NSDUH) showed that an estimated 22.6 million, or 8.9% of Americans, aged 12 or older, were current or past month illicit drug users, The survey showed that just behind the 7 million people who had used marijuana, 5.1 million had used pain relievers. It has also been shown that only one in 6 or 17.3% of users of non-therapeutic opioids indicated that they received the drugs through a prescription from one doctor. The escalating use of therapeutic opioids shows hydrocodone topping all prescriptions with 136.7 million prescriptions in 2011, with all narcotic analgesics exceeding 238 million prescriptions. It has also been illustrated that opioid analgesics are now responsible for more deaths than the number of deaths from both suicide and motor vehicle crashes, or deaths from cocaine and heroin combined. A significant relationship exists between sales of opioid pain relievers and deaths. The majority of deaths (60%) occur in patients when they are given prescriptions based on prescribing guidelines by medical boards, with 20% of deaths in low dose opioid therapy of 100 mg of morphine equivalent dose or less per day and 40% in those receiving morphine of over 100 mg per day. In comparison, 40% of deaths occur in individuals abusing the drugs obtained through multiple prescriptions, doctor shopping, and drug diversion. The purpose of this comprehensive review is to describe various aspects of crisis of opioid use in the United States. The obstacles that must be surmounted are primarily inappropriate prescribing patterns, which are largely based on a lack of knowledge, perceived safety, and inaccurate belief of undertreatment of pain.


Post-traumatic hypoxia exacerbates neuronal cell death in the hippocampus.
Hypoxia frequently occurs in patients with traumatic brain injury (TBI) and is associated with increased morbidity and mortality. This study examined the effects of immediate or delayed post-traumatic hypoxia (fraction of inspired oxygen [FiO(2)] 11%) on acute neuronal degeneration and long-term neuronal survival in hippocampal fields after moderate fluid percussion injury in rats. In Experiment 1, hypoxia was induced for 15 or 30 min alone or immediately following TBI. In Experiments 2 and 3, 30 min of hypoxia was induced immediately after TBI or delayed until 60 min after TBI. In Experiment 1, acute neurodegeneration was evaluated in the hippocampal fields 24 h after insults using Fluoro-Jade staining and stereological quantification. During hypoxia alone, or in combination with TBI, mean arterial blood pressure was significantly reduced by approximately 30%, followed by a rapid return to normal values upon return to pre-injury FiO(2). Hypoxia alone failed to cause hippocampal neuronal degeneration when measured at 24 h after insult. TBI alone resulted in neuronal degeneration in each ipsilateral hippocampal field, predominantly in CA2-CA3 and the dentate gyrus. Compared to TBI alone, TBI plus immediate hypoxia for either 15 or 30 min significantly increased neuronal loss in most ipsilateral hippocampal fields and in the contralateral hilus and dentate gyrus. In Experiment 2, TBI plus hypoxia delayed 30 min significantly increased degeneration only in ipsilateral CA2-CA3. In Experiment 3, 30 min of immediate hypoxia significantly reduced the numbers of surviving neurons in the CA3 at 14 days after TBI. The greatly increased vulnerability in all hippocampal fields by immediate 30 min post-traumatic hypoxia provides a relevant model of TBI complicated with hypoxia/hypotension. These data underscore the significance of the secondary insult, the necessity to better characterize the range of injuries experienced by the TBI patient, and the importance of strictly avoiding hypoxia in the early management of TBI patients.


[Obtention with polymerase chain reaction (PCR) of gene rbcL in the scrophulariaceae holoparasite Lathraea clandestina L. Comparison with autotroph Digitalis purpurea L. and Melampyrum pratense L].
Amyloplastid DNA was extracted from the Scrophulariaceae holoparasite Lathraea clandestina L. and then purified. An rbcL gene amplification was performed using polymerase chain reaction. Two regions of well conserved sequences in Tobacco, Spinach and Maize rbcL gene have been used as primers. PCR yields a sequence of about 1,230 base pairs, almost nine tenths of the rbcL coding region. Identical results were obtained with plastid DNAs from Tobacco and two other species of Scrophulariaceae: the non-parasitic Digitalis purpurea L. and the hemiparasite Melampyrum pratense L. PCR products were digested with BamHI restriction enzyme and no changes were shown in the localization of the restriction site whatever the species assayed. Identical restriction patterns were also observed with Tobacco and Digitalis PCR products restricted with PstI and KpnI, whereas Melampyrum and Lathraea exhibited a different restriction pattern with PstI. So, despite slight differences, some analogies between Lathraea and Digitalis or Tobacco gene were evidenced. Cloning and sequencing of these PCR products could give a more accurate response to the following question: to what extent have changes occurred in the rbcL gene in a plant which lacks chlorophyll?


Dual role for the zeste-white3/shaggy-encoded kinase in mesoderm and heart development of Drosophila.
A Drosophila homolog of the serine/threonine kinase GSK-3 beta, encoded by the zest-white3/shaggy gene (zw3), has been implicated as a maternally provided antagonist of zygotic signaling by the secreted segmentation gene wingless (wg). The wg signal apparently causes a spatially localized inhibition of the ubiquitous repressor function of zw3. This double negative mechanism of signal transduction has been shown to mediate the patterning function of Wg in a number of developmental processes. Although wg is absolutely required for specifying the heart progenitors within the mesoderm of Drosophila, the role of zw3 in this process has been unclear. Here, we present evidence that zw3 has a dual role in mesoderm development: (1) zw3 acts as an antagonist in cardiogenic wg signal transduction, and (2) zw3 also seems to be required to promote positively the formation of a larger mesodermal region, the tinman- and dpp-dependent "dorsal mesoderm," which is a prerequisite not only for cardiogenesis, but also for visceral mesoderm formation. We also demonstrate that a recently identified proximal component of the wg cascade, which is a transcription factor encoded by pangolin/dTCF (dTCF), also seems to mediate wg-dependent cardiogenesis. Further, we present evidence that Notch (N), which opposes wg signaling in other situations, is unlikely to be directly involved in the cardiogenic wg pathway, but seems to have multiple other myogenic functions, one of which is to inhibit mesoderm differentiation altogether, when overexpressed as a constitutively active form.


Usefulness of a stool to stabilize dental chairs for cardiopulmonary resuscitation (CPR).
Cardiopulmonary resuscitation (CPR) requires immediate start of manual chest compression (MCC) and defibrillation as soon as possible. During dental surgery, CPR could be started in the dental chair considering difficulty to move the patient from the dental chair to the floor. However, all types of dental chairs are not stable for MCC. We previously developed a procedure to stabilize a dental chair by using a stool. EUROPEAN RESUSCITATION COUNCIL (ERC) guideline 2015 adopted our procedure when cardiac arrest during dental surgery. The objective of this study was to verify the efficacy of a stool as a stabilizer in different types of dental chairs. Three health care providers participated in this study, and 8 kinds of dental chairs were examined. MCC were performed on a manikin that was laid on the backrest of a dental chair. A stool was placed under the backrest to stabilize the dental chair. The vertical displacement of the backrest by MCC was recorded by a camcorder and measured by millimeter. Next, the vertical displacement of the backrest by MCC were compared between with and without a stool. In all 8 dental chairs, the method by using a stool significantly reduced the vertical displacements of the backrest by during MCC. The reduction ratio (mean [interquartile range]) varied between nearly 27 [20] and 87 [5] %. In the largest stabilization case, the displacement was 3.5 [0.5] mm with a stool versus 26 [5.5] mm without a stool (p < 0.001). Our procedure to stabilize dental chairs by using a stool reduced the displacement of a backrest against MCC in all chairs. Effective MCC could be performed in dental chairs by using a stool when sudden cardiac arrest occurs during dental surgery.


Risk Factors for Developing Postlumbar Puncture Headache: A Case-Control Study.
Postlumbar puncture headache (PLPH) is a common adverse event after lumbar puncture. The specific cause for headache remains uncertain but is assumed to be cerebrospinal fluid leakage. The headache affects the patient, family, and workplace as well as society in general. The aim of this study was to identify risk factors associated with the development of PLPH. This quantitative case-control study included 285 patients having undergone diagnostic lumbar puncture at a department of infectious diseases during 2015 to 2016. We examined blood glucose and blood albumin levels as well as systolic, diastolic, and mean arterial blood pressure. Statistical analysis included Student t test, χ, and logistic regression. Compared with the control group, patients with PLPH tended to have a lower glucose level (5.7 vs 6.4 mmol/L), lower systolic pressure (126 vs 137 mm Hg), and lower mean arterial blood pressure (90 vs 96 mm Hg). Logistic regression analysis showed that lower systolic blood pressure (≤126 mm Hg) significantly increased the risk of PLPH (odds ratio, 0.977 [95% confidence interval, 0.957-0.998]) along with age younger than 40 years (odds ratio, 0.954 [95% confidence interval, 0.935-0.973]). Patients with a lower systolic blood pressure had a statistically significantly increased risk of PLPH. Furthermore, younger age was also associated with an increased risk of PLPH.


Transnational telepathology consultations using a basic digital microscope: experience in the Italy-Slovenjia INTERREG project "patient without borders".
In recent years, a number of technological advancements started to modify the long standing appearance and functionalities of traditional optical microscopes used in Pathology and other medical fields. In fact, at present many new tools for microscopical visualization exist that are based on digital imaging, robotization, and remote communication. Such tools are typically adopted in activities ranging from education to telediagnosis to remote consultation. Present paper describes the features of a basic digital microscope that has been tested to verify its performance for occasional remote consultation inside an international project between Italy and Slovenija, funded by Interreg initiative of the European Regional Development Fund. The system is composed by a pair of digital microscopes (Leica DMD108, Leitz Microsystems, Wetzlar, Germany) associated to a high resolution videoconferencing systems (Tandberg 990, Lysaker, Norway). The systems are connected through the Internet. Sixty histology and cytology cases have been collaboratively diagnosed between two Pathology Institutes to verify the diagnostic performance of the system, regarding the image quality point of view as well as time needed for diagnosis. The system has also been tested for compatibility with standard videoconferencing software. No discrepancies between local and remote diagnoses have been identified, with diagnosis time reasonably close to typical microscope observation times. Time needed for most operations is not far from that needed on a traditional microscope, except for startup. The system can be considered usable as a standard microscope, and also for occasional remote consultations.