Timing of the onset of acute respiratory distress syndrome: a population-based study.
Many patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) have had recent healthcare interventions prior to developing ALI/ARDS. To determine the timing of ALI/ARDS onset in relation to hospital admission and other healthcare interventions. We conducted a population-based observational cohort study with a validated electronic surveillance tool, and identified patients with possible ALI/ARDS among critically ill adults at Mayo Clinic hospitals that provide critical care services for Olmsted County, Minnesota, in 2006. Trained investigators independently reviewed electronic medical records and confirmed the presence and timing of ALI/ARDS based on the American-European consensus definition. Of 124 episodes of ALI in 118 patients, only 5 did not fulfill the ARDS criteria. The syndrome developed a median 30 hours (IQR 10-82 h) after hospital admission in 79 patients (67%). ARDS was present on admission in 39 patients (33%), of whom 14 had recent hospitalization, 6 were transferred from nursing homes, and 3 had recent out-patient contact (1 antibiotic prescription, 1 surgical intervention, and 1 chemotherapy). Only 16 ARDS patients (14%) did not have known recent contact with a healthcare system. Compared to ARDS on admission, hospital-acquired ARDS was more likely to occur in surgery patients (54% vs 15%, P < .001), and had longer adjusted hospital stay (mean difference 8.9 d, 95% CI 0.3-17.4, P = .04). ARDS in the community most often develops either during hospitalization or in patients who recently had contact with a healthcare system. These findings have important implications for potential preventive strategies.


Stromally expressed β-catenin modulates Wnt9b signaling in the ureteric epithelium.
The mammalian kidney undergoes cell interactions between the epithelium and mesenchyme to form the essential filtration unit of the kidney, termed the nephron. A third cell type, the kidney stroma, is a population of fibroblasts located in the kidney capsule, cortex and medulla and is ideally located to affect kidney formation. We found β-catenin, a transcriptional co-activator, is strongly expressed in distinctive intracellular patterns in the capsular, cortical, and medullary renal stroma. We investigated β-catenin function in the renal stroma using a conditional knockout strategy that genetically deleted β-catenin specifically in the renal stroma cell lineage (β-cats-/-). β-cats-/- mutant mice demonstrate marked kidney abnormalities, and surprisingly we show β-catenin in the renal stroma is essential for regulating the condensing mesenchyme cell population. We show that the population of induced mesenchyme cells is significantly reduced in β-cats-/- mutants and exhibited decreased cell proliferation and a specific loss of Cited 1, while maintaining the expression of other essential nephron progenitor proteins. Wnt9b, the key signal for the induction of nephron progenitors, was markedly reduced in adjacent ureteric epithelial cells in β-cats-/-. Analysis of Wnt9b-dependent genes in the neighboring nephron progenitors was significantly reduced while Wnt9b-independent genes remained unchanged. In contrast mice overexpressing β-catenin exclusively in the renal stroma demonstrated massive increases in the condensing mesenchyme population and Wnt9b was markedly elevated. We propose that β-catenin in the renal stroma modulates a genetic program in ureteric epithelium that is required for the induction of nephron progenitors.


Success rate of cervical exploration for hyperparathyroidism.
In a three-year experience with 361 patients who underwent exploratory surgery for hyperparathyroidism, the success rate for primary cervical operations was 95% and that for secondary operations (cervical and mediastinal) was 62%; for all operations, it was 94%. The three most important factors in determining successful cervical exploration of the parathyroid glands are correct preoperative diagnosis, meticulous surgical technique, and accurate determination of abnormal locations of pathologic conditions. Ancillary studies or techniques for preoperative localization of parathyroid tissue were utilized in only a few patients who had unusually complicated problems. The results of this study suggest that, because of cost, time involvement, and potential risk, selective venous sampling with radioimmunoassay of parathyroid hormone or arteriography or both should be reserved for complicated problems and for patients being considered for a second or third exploratory operation.


Ventral and dorsal streams processing visual motion perception (FDG-PET study).
Earlier functional imaging studies on visually induced self-motion perception (vection) disclosed a bilateral network of activations within primary and secondary visual cortex areas which was combined with signal decreases, i.e., deactivations, in multisensory vestibular cortex areas. This finding led to the concept of a reciprocal inhibitory interaction between the visual and vestibular systems. In order to define areas involved in special aspects of self-motion perception such as intensity and duration of the perceived circular vection (CV) or the amount of head tilt, correlation analyses of the regional cerebral glucose metabolism, rCGM (measured by fluorodeoxyglucose positron-emission tomography, FDG-PET) and these perceptual covariates were performed in 14 healthy volunteers. For analyses of the visual-vestibular interaction, the CV data were compared to a random dot motion stimulation condition (not inducing vection) and a control group at rest (no stimulation at all). Group subtraction analyses showed that the visual-vestibular interaction was modified during CV, i.e., the activations within the cerebellar vermis and parieto-occipital areas were enhanced. The correlation analysis between the rCGM and the intensity of visually induced vection, experienced as body tilt, showed a relationship for areas of the multisensory vestibular cortical network (inferior parietal lobule bilaterally, anterior cingulate gyrus), the medial parieto-occipital cortex, the frontal eye fields and the cerebellar vermis. The "earlier" multisensory vestibular areas like the parieto-insular vestibular cortex and the superior temporal gyrus did not appear in the latter analysis. The duration of perceived vection after stimulus stop was positively correlated with rCGM in medial temporal lobe areas bilaterally, which included the (para-)hippocampus, known to be involved in various aspects of memory processing. The amount of head tilt was found to be positively correlated with the rCGM of bilateral basal ganglia regions responsible for the control of motor function of the head. Our data gave further insights into subfunctions within the complex cortical network involved in the processing of visual-vestibular interaction during CV. Specific areas of this cortical network could be attributed to the ventral stream ("what" pathway) responsible for the duration after stimulus stop and to the dorsal stream ("where/how" pathway) responsible for intensity aspects.


Multi-locus sequence typing: a tool for global epidemiology.
The characterization of pathogenic isolates plays a pivotal role in the epidemiology of infectious diseases, generating the information necessary for identifying, tracking, and intervening against disease outbreaks. In 1998 multi-locus sequence typing (MLST) was proposed as a nucleotide sequence-based approach that could be applied to many bacterial pathogens. It combined developments in high-throughput sequencing and bioinformatics with established population genetics techniques to provide a portable, reproducible, and scalable typing system that reflected the population and evolutionary biology of bacterial pathogens. MLST schemes have been developed for a variety of procaryotic and eucaryotic pathogens and the data generated have contributed to both epidemiological surveillance and fundamental studies of pathogen biology.


RTEC-intrinsic IL-17-driven inflammatory circuit amplifies antibody-induced glomerulonephritis and is constrained by Regnase-1.
Antibody-mediated glomerulonephritis (AGN) is a clinical manifestation of many autoimmune kidney diseases for which few effective treatments exist. Chronic inflammatory circuits in renal glomerular and tubular cells lead to tissue damage in AGN. These cells are targeted by the cytokine IL-17, which has recently been shown to be a central driver of the pathogenesis of AGN. However, surprisingly little is known about the regulation of pathogenic IL-17 signaling in the kidney. Here, using a well-characterized mouse model of AGN, we show that IL-17 signaling in renal tubular epithelial cells (RTECs) is necessary for AGN development. We also show that Regnase-1, an RNA binding protein with endoribonuclease activity, is a negative regulator of IL-17 signaling in RTECs. Accordingly, mice with a selective Regnase-1 deficiency in RTECs exhibited exacerbated kidney dysfunction in AGN. Mechanistically, Regnase-1 inhibits IL-17-driven expression of the transcription factor IκBξ and, consequently, its downstream gene targets, including Il6 and Lcn2. Moreover, deletion of Regnase-1 in human RTECs reduced inflammatory gene expression in a IκBξ-dependent manner. Overall, these data identify an IL-17-driven inflammatory circuit in RTECs during AGN that is constrained by Regnase-1.


Short-term global motion adaptation induces a compression in the subjective duration of dynamic visual events.
Apparent duration can be manipulated in a local region of visual field by long-term adaptation to motion or flicker (Johnston, Arnold, & Nishida, 2006). These effects show narrow spatial tuning (Ayhan, Bruno, Nishida, & Johnston, 2009), as well as retinotopic position dependency (Bruno, Ayhan, & Johnston, 2010), supporting early locus in the visual pathway. Here, we introduce a new effect using RDK as a short-term visual adaptor and demonstrate that a brief, subsecond range adaptation induces a significant subjective duration compression (∼10%) on a subsequently presented test stimulus (RDK pattern) only for global motion patterns drifting at 50% motion coherence but not for those drifting at 0% coherence, suggesting a higher level area as a source of origin. In another set of experiments using a plaid stimulus as the adaptor and gratings as the tests, we report again a significant duration compression following a brief motion adaptation, although the effect does not seem to be consistently selective for a particular direction of the standard test relative to that of the plaid adaptor (two-dimensional motion) or its components (one-dimensional motion). Finally, we conduct an experiment using shutter glasses and find that the effects of a short-term adaptor presented monocularly to one eye transfer to the nonadapted eye, providing evidence for the interocular transfer. In a series of control experiments, we also show that the duration effects cannot be explained by adaptation-induced changes in perceived speed, perceived onset-and-offset, and attentional resource allocation. Overall, the duration compression effect requiring motion coherence in RDK, persisting in plaid stimulus, and showing interocular transfer imply explicit genuine mechanisms mediating duration effects in the higher level motion areas.


Cytokine production by infrapatellar fat pad can be stimulated by interleukin 1β and inhibited by peroxisome proliferator activated receptor α agonist.
Infrapatellar fat pad (IPFP) might be involved in osteoarthritis (OA) by production of cytokines. It was hypothesised that production of cytokines is sensitive to environmental conditions. To evaluate cytokine production by IPFP in response to interleukin (IL)1β and investigate the ability to modulate this response with an agonist for peroxisome proliferator activated receptor α (PPARα), which is also activated by lipid-lowering drugs such as fibrates. Cytokine secretion of IPFP was analysed in the medium of explant cultures of 29 osteoarthritic patients. IPFP (five donors) and synovium (six donors) were cultured with IL-1β and PPARα agonist Wy14643. Gene expression of IL-1β, monocyte chemoattractant protein (MCP1), (IL-6, tumour necrosis factor (TNF)α, leptin, vascular endothelial growth factor (VEGF), IL-10, prostaglandin-endoperoxide synthase (PTGS)2 and release of TNFα, MCP1 and prostaglandin E(2) were compared with unstimulated IPFP and synovium explants. IPFP released large amounts of inflammatory cytokines, adipokines and growth factors. IL-1β increased gene expression of PTGS2, TNFα, IL-1β, IL-6 and VEGF and increased TNFα release in IPFP. MCP1, leptin, IL-10 gene expression and MCP1, leptin and PGE(2) release did not increase significantly. Synovium responded to IL-1β similarly to IPFP, except for VEGF gene expression. Wy14643 decreased gene expression of PTGS2, IL-1β, TNFα, MCP1, VEGF and leptin in IPFP explants and IL-1β, TNFα, IL-6, IL-10 and VEGF in synovium that responded to IL-1β. IPFP is an active tissue within the joint. IPFP cytokine production is increased by IL-1β and decreased by a PPARα agonist. The effects were similar to effects seen in synovium. Fibrates may represent a potential disease-modifying drug for OA by modulating inflammatory properties of IPFP and synovium.


Antigenicity and immunogenicity of allogeneic retinal transplants.
The transplantation of neuronal cells and tissues represents a promising approach for the treatment of incurable neurodegenerative diseases. Indeed, it has been reported recently that retinal transplantation can rescue photoreceptor cells and delay age-related changes in various retinal layers in rodents. However, retinal grafts deteriorate progressively after placement in recipients' eyes. Here we investigated whether a host's immune response elicited toward the graft contributes to its deterioration. Using an ELISA spot assay, we measured T cell responses to retinal tissues placed in the vitreous cavity of syngeneic and allogeneic mice. We found that allogeneic retinas induced potent alloimmune responses mediated by T cells secreting type 1 cytokines (IFN-gamma and IL-2). No response was found in mice engrafted with syngeneic retinas. In addition, all syngeneic retinal grafts displayed no signs of tissue damage (at 55 days), while the majority of allogeneic retinas deteriorated as early as 12 days after placement. Next, we showed that anti-donor responses occurred within two phenotypically and functionally distinct T cell subsets: CD4+ T cells secreting IL-2 and CD8+ T cells producing IFN-gamma. Importantly, CD4+ T cells were necessary and sufficient to cause graft deterioration, while CD8+ T cells did not contribute to this process.


Occurrence of Triclocarban and Triclosan in an Agro-ecosystem Following Application of Biosolids.
Triclocarban (TCC) and triclosan (TCS), two of the most commonly used antimicrobial compounds, can be introduced into ecosystems by applying wastewater treatment plant biosolids to agricultural fields. Concentrations of TCC and TCS were measured in different trophic levels within a terrestrial food web encompassing land-applied biosolids, soil, earthworms (Lumbricus), deer mice (Peromyscus maniculatus), and eggs of European starlings (Sturnus vulgaris) and American kestrels (Falco sparverius) at an experimental site amended with biosolids for the previous 7 years. The samples from this site were compared to the same types of samples from a reference (biosolids-free) agricultural site. Inter-site comparisons showed that concentrations of both antimicrobials were higher on the experimental site in the soil, earthworms, mice (livers), and European starling eggs, but not American kestrel eggs, compared to the control site. Inter-species comparisons on the experimental site indicated significantly higher TCC concentrations in mice (TCC: 12.6-33.3 ng/g) and in starling eggs (TCC: 15.4-31.4 ng/g) than in kestrel eggs (TCC: 3.6 ng/g). Nesting success of kestrels only was significantly lower on the experimental site compared to the reference site due to nest abandonment. This study demonstrates that biosolids-derived TCC and TCS are present throughout the terrestrial food web, including secondary (e.g., starlings) and tertiary (i.e., kestrels) consumers, after repeated, long-term biosolids application.


Suctioning habits in the delivery room and the influence on postnatal adaptation - a video analysis.
To determine how often infants are suctioned during delivery and how it affects the neonate. Single-center analysis of video-recorded delivery room management after c-section from January 2012 until April 2013. Time point, duration, and frequency of suctioning in term and preterm newborns were analyzed along with vital parameters (heart rate (HR) and saturation values). Three hundred forty-six videos were analyzed. Twenty-three percent of term and 66% of preterm newborns were suctioned. Newborns were suctioned up to 14 times; total duration spent for suctioning was between 2 and 154 s. Suctioning before face mask application occurred in 31% of the suctioned newborns requiring respiratory support. No severe bradycardia (<60 bpm) was noticed. Suctioning did not have an effect on HR and saturation in preterm infants but was associated with significantly higher HR in term infants requiring respiratory support. Term infants who did not require respiratory support showed significantly higher saturation values at 3, 5, 6, 7, 8, 9, and 10 min if they were not suctioned. Suctioning of newborns in the delivery room does not adhere to recommendations of international guidelines. However, previously described side effects of suctioning could not be confirmed.


Characteristics of a novel treatment system for linear accelerator-based stereotactic radiosurgery.
The purpose of this study is to characterize the dosimetric properties and accuracy of a novel treatment platform (Edge radiosurgery system) for localizing and treating patients with frameless, image-guided stereotactic radiosurgery (SRS) and stereotactic body radiotherapy (SBRT). Initial measurements of various components of the system, such as a comprehensive assessment of the dosimetric properties of the flattening filter-free (FFF) beams for both high definition (HD120) MLC and conical cone-based treatment, positioning accuracy and beam attenuation of a six degree of freedom (6DoF) couch, treatment head leakage test, and integrated end-to-end accuracy tests, have been performed. The end-to-end test of the system was performed by CT imaging a phantom and registering hidden targets on the treatment couch to determine the localization accuracy of the optical surface monitoring system (OSMS), cone-beam CT (CBCT), and MV imaging systems, as well as the radiation isocenter targeting accuracy. The deviations between the percent depth-dose curves acquired on the new linac-based system (Edge), and the previously published machine with FFF beams (TrueBeam) beyond D(max) were within 1.0% for both energies. The maximum deviation of output factors between the Edge and TrueBeam was 1.6%. The optimized dosimetric leaf gap values, which were fitted using Eclipse dose calculations and measurements based on representative spine radiosurgery plans, were 0.700 mm and 1.000 mm, respectively. For the conical cones, 6X FFF has sharper penumbra ranging from 1.2-1.8 mm (80%-20%) and 1.9-3.8 mm (90%-10%) relative to 10X FFF, which has 1.2-2.2mm and 2.3-5.1mm, respectively. The relative attenuation measurements of the couch for PA, PA (rails-in), oblique, oblique (rails-out), oblique (rails-in) were: -2.0%, -2.5%, -15.6%, -2.5%, -5.0% for 6X FFF and -1.4%, -1.5%, -12.2%, -2.5%, -5.0% for 10X FFF, respectively, with a slight decrease in attenuation versus field size. The systematic deviation between the OSMS and CBCT was -0.4 ± 0.2 mm, 0.1± 0.3mm, and 0.0 ± 0.1 mm in the vertical, longitudinal, and lateral directions. The mean values and standard deviations of the average deviation and maximum deviation of the daily Winston-Lutz tests over three months are 0.20 ± 0.03 mm and 0.66 ± 0.18 mm, respectively. Initial testing of this novel system demonstrates the technology to be highly accurate and suitable for frameless, linac-based SRS and SBRT treatment.


SMA Selectively Codes the Active Accumulation of Temporal, Not Spatial, Magnitude.
Estimating duration depends on the sequential integration (accumulation) of temporal information in working memory. Using fMRI, we directly compared the accumulation of information in temporal versus spatial domains. Participants estimated either the duration or distance of the dynamic trajectory of a moving dot or, in a control condition, a static line stimulus. Comparing the duration versus distance of static lines activated an extensive cortico-striatal network. By contrast, comparing the duration versus distance of dynamic trajectories, both of which required sequential integration of information, activated SMA alone. Indeed, activity in SMA, as well as right inferior occipital cortex, increased parametrically as a function of stimulus duration and also correlated with individual differences in the propensity to overestimate stimulus duration. By contrast, activity in primary visual cortex increased parametrically as a function of stimulus distance. Crucially, a direct comparison of the parametric responses to duration versus distance revealed that activity in SMA increased incrementally as a function of stimulus duration but not as a function of stimulus distance. Collectively, our results indicate that SMA responds to the active accumulation of information selectively in the temporal domain.


[A new diarrhea pathogen: entero-SLTs-producing and invasive Escherichia coli was over-looked as normal flora E. coli].
In clinical laboratories of Beijing, China, no known entero-pathogen but almost pure E. coli strains could be isolated from more than 60% fecal samples of diarrheal patients, which had been recognized as normal flora E. coli and dismissed. We suspected that some of the so-called normal flora E. coli strains might be virulent. To prove our idea. We collected 172 strains of E. coli isolated from diarrheal patients from whom no other enteric bacterial pathogens had been identified, including EPEC, EIEC, ETEC. With plasmid DNA analysis, Hep-2 cell adherence assay and 10 DNA probe hybridization, we found that the so-called normal flora E. coli was abnormal, 44% of them were virulent, of which 16 (9.3) were EHEC, 8 EPEC (4.7%), 11 EAggEC (6.8%). Fifty-four of 172 strains were hybridized with INV and SLT1 or SLT2 probes, which had never been reported. These strains could invade Hep-2 cells, but were lack of ipaB gene, a key gene of invasiveness gene cluster of Shigella species and EIEC. The aggregative adherence to Hep-2 cells was observed, but the strains were not hybridized with EAggEC specific DNA probe. The purified toxin protein and cell filtrate were toxic to vero cells. Based on the data obtained, we believed that this is a new category of diarrhea-genic E. coli, named as entero-SLTs-producing and invasive E.coli (ESIEC). ESIEC occupied 31.4% of the strains tested, the isolation of it was probably higher than those of ETEC, EPEC in P R China.


Supramolecular photocatalysis: combining confinement and non-covalent interactions to control light initiated reactions.
Using non-bonding interactions to control photochemical reactions requires an understanding of not only thermodynamics and kinetics of ground state and excited state processes but also the intricate interactions that dictate the dynamics within the system of interest. This review is geared towards a conceptual understanding of how one can control the reactivity and selectivity in the excited state by employing confinement and non-covalent interactions. Photochemical reactivity of organic molecules within confined containers and organized assemblies as well as organic templates that interact through H-bonding and/or cation-carbonyl/cation-π interactions is reviewed with an eye towards understanding supramolecular effects and photocatalysis.


Genetic aspects of adolescent idiopathic scoliosis in a family with multiple affected members: a research article.
The etiology of idiopathic scoliosis remains unknown and different factors have been suggested as causal. Hereditary factors can also determine the etiology of the disease; however, the pattern of inheritance remains unknown. Autosomal dominant, X-linked and multifactorial patterns of inheritances have been reported. Other studies have suggested possible chromosome regions related to the etiology of idiopathic scoliosis. We report the genetic aspects of and investigate chromosome regions for adolescent idiopathic scoliosis in a Brazilian family. Evaluation of 57 family members, distributed over 4 generations of a Brazilian family, with 9 carriers of adolescent idiopathic scoliosis. The proband presented a scoliotic curve of 75 degrees, as determined by the Cobb method. Genomic DNA from family members was genotyped. Locating a chromosome region linked to adolescent idiopathic scoliosis was not possible in the family studied. While it was not possible to determine a chromosome region responsible for adolescent idiopathic scoliosis by investigation of genetic linkage using microsatellites markers during analysis of four generations of a Brazilian family with multiple affected members, analysis including other types of genomic variations, like single nucleotide polymorphisms (SNPs) could contribute to the continuity of this study.


Cbl-b regulates the CD28 dependence of T-cell activation.
Whereas co-stimulation of the T-cell antigen receptor (TCR) and CD28 triggers T-cell activation, stimulation of the TCR alone may result in an anergic state or T-cell deletion, both possible mechanisms of tolerance induction. Here we show that T cells that are deficient in the adaptor molecule Cbl-b (ref. 3) do not require CD28 engagement for interleukin-2 production, and that the Cbl-b-null mutation (Cbl-b(-/-)) fully restores T-cell-dependent antibody responses in CD28-/- mice. The main TCR signalling pathways, such as tyrosine kinases Zap-70 and Lck, Ras/mitogen-activated kinases, phospholipase Cgamma-1 and Ca2+ mobilization, were not affected in Cbl-b(-/-) T cells. In contrast, the activation of Vav, a guanine nucleotide exchange factor for Rac1/Rho/CDC42, was significantly enhanced. Our findings indicate that Cbl-b may influence the CD28 dependence of T-cell activation by selectively suppressing TCR-mediated Vav activation. Mice deficient in Cbl-b are highly susceptible to experimental autoimmune encephalomyelitis, suggesting that the dysregulation of signalling pathways modulated by Cbl-b may also contribute to human autoimmune diseases such as multiple sclerosis.


Effects of Mindful Parenting Training on Clinical Symptoms in Children with Attention Deficit Hyperactivity Disorder and Parenting Stress: Randomized Controlled Trial.
Children with attention deficit hyperactivity disorder (ADHD) are at risk of impairment in multiple domains. This study aimed to investigate the effectiveness of mindful parenting training in reducing clinical symptoms in children with ADHD and parenting stress of their parents. This randomized clinical trial was conducted on 2 groups (experimental and control) in 3 phases (pretest, posttest, and 8 weeks' follow-up). Sixty children with ADHD, who had been referred by the child psychiatrist in the Iranian city of Kashan in the second half of the year 2016, were selected along with their mothers. The mothers were assigned to one of the 2 groups via permuted blocked randomization. The mothers completed the parenting stress index-short form (PSI-SF 36) and the Swanson, Nolan, and Pelham Parent and Teacher rating scale (SNAP-IV). All the children in both groups received pharmacotherapy with either risperidone or Ritalin. The intervention group received 8 sessions (1 session each week, each session lasting 90 minutes) of mindful parenting training based on the Kabat-Zinn protocol. The data were analyzed using SPSS, version 20, via the t test, χ2 test, repeated measures analysis of variance, and nonparametric Friedman test. This study showed a reduction in parenting stress, negative parent-child interactions, and children's problematic characteristics in the mindful parenting training group compared with the control group in the posttest and follow-up. Our results also demonstrated a significant improvement in ADHD symptoms in the experimental group by comparison with the control group in the posttest and follow-up. Mindful parenting training was effective in reducing parenting stress and ADHD symptoms in our intervention group. Trial Registration Number: IRCT2016021026505N1.


Maternal common mental disorders and infant development in Ethiopia: the P-MaMiE Birth Cohort.
Chronicity and severity of early exposure to maternal common mental disorders (CMD) has been associated with poorer infant development in high-income countries. In low- and middle-income countries (LAMICs), perinatal CMD is inconsistently associated with infant development, but the impact of severity and persistence has not been examined. A nested population-based cohort of 258 pregnant women was identified from the Perinatal Maternal Mental Disorder in Ethiopia (P-MaMiE) study, and 194 (75.2%) were successfully followed up until the infants were 12 months of age. Maternal CMD was measured in pregnancy and at two and 12 months postnatal using the WHO Self-Reporting Questionnaire, validated for use in this setting. Infant outcomes were evaluated using the Bayley Scales of Infant Development. Antenatal maternal CMD symptoms were associated with poorer infant motor development (β^ -0.20; 95% CI: -0.37 to -0.03), but this became non-significant after adjusting for confounders. Postnatal CMD symptoms were not associated with any domain of infant development. There was evidence of a dose-response relationship between the number of time-points at which the mother had high levels of CMD symptoms (SRQ ≥ 6) and impaired infant motor development (β^ = -0.80; 95%CI -2.24, 0.65 for ante- or postnatal CMD only, β^ = -4.19; 95%CI -8.60, 0.21 for ante- and postnatal CMD, compared to no CMD; test-for-trend χ(2)13.08(1), p < 0.001). Although this association became non-significant in the fully adjusted model, the β^ coefficients were unchanged indicating that the relationship was not confounded. In multivariable analyses, lower socio-economic status and lower infant weight-for-age were associated with significantly lower scores on both motor and cognitive developmental scales. Maternal experience of physical violence was significantly associated with impaired cognitive development. The study supports the hypothesis that it is the accumulation of risk exposures across time rather than early exposure to maternal CMD per se that is more likely to affect child development. Further investigation of the impact of chronicity of maternal CMD upon child development in LAMICs is indicated. In the Ethiopian setting, poverty, interpersonal violence and infant undernutrition should be targets for interventions to reduce the loss of child developmental potential.


Molecular characterization of Bacillus pasteurii UreE, a metal-binding chaperone for the assembly of the urease active site.
The present study describes the cloning, isolation, and thorough biochemical characterization of UreE from Bacillus pasteurii, a novel protein putatively involved in the transport of Ni in the urease assembly process. A DNA fragment of the B. pasteurii urease operon, containing all four accessory genes (ureE, ureF, ureG, and ureD) required for the incorporation of Ni ions into the active site of urease, was cloned, sequenced, and analyzed. B. pasteurii ureE was cloned, and the UreE protein (BpUreE) was over-expressed and purified to homogeneity. The identity of the recombinant protein was determined by N- and C-terminal sequencing and by mass spectrometry. BpUreE has a chain length of 147 amino acids, and features a p I value of 4.7. As isolated, BpUreE contains one Zn(II) ion per dimer, while no Ni(II) is present, as shown by mass spectrometry and atomic absorption spectroscopy. BpUreE behaves as a dimer independently of the presence of Zn(II), as shown by gel filtration and mass spectrometry. Paramagnetic NMR spectroscopy on concentrated (2 mM) UreE solutions reveals a one Ni atom per tetramer stoichiometry, with the Ni(II) ion bound to histidines in an octahedral coordination environment. BpUreE has a high sequence similarity with UreE proteins isolated from different biological sources, while no sequence homology is observed with proteins belonging to different classes. In particular, BpUreE is most similar to UreE from Bacillus halodurans (55% identity). A multiple sequence alignment reveals the presence of four strictly conserved residues (Leu55, Gly97, Asn98, His100; BpUreE numbering), in addition to position 115, conservatively occupied by an Asp or a Glu residue. Several secondary structure elements, including a betaalphabetabetaalphabeta "ferredoxin-like" motif, are highly conserved throughout the UreE sequences.


Anti-inflammatory activity of quercetin and isoquercitrin in experimental murine allergic asthma.
Eosinophils and cytokines are implicated in the pathogenesis of allergic diseases. In the present study, we investigate the anti-inflammatory effect of quercetin and isoquercitrin in a murine model of asthma. BALB/c mice were immunized (ovalbumin/aluminum hydroxide, s. c.), followed by two intranasal ovalbumin challenges. From day 18 to day 22 after the first immunization, the mice received daily gavages of isoquercitrin (15 mg/kg) or quercetin (10 mg/kg). Dexamethasone (1 mg/kg, s. c.) was administered as a positive control. Leucocytes were analyzed in bronchoalveolar lavage fluid (BALF), blood and pulmonary parenchyma at 24 h after the last ovalbumin challenge. Interleukin-5 (IL-5) was analyzed in BALF and lung homogenates. In animals receiving isoquercitrin or quercetin, eosinophil counts were lower in the BALF, blood and lung parenchyma. Neutrophil counts in blood and IL-5 levels in lung homogenate were lower only in isoquercitrin-treated mice. No alterations in mononuclear cell numbers were observed. Quercetin and isoquercitrin are effective eosinophilic inflammation suppressors, suggesting a potential for treating allergies.


Synthesis of granulocyte colony-stimulating factor and its requirement for terminal divisions in chronic myelogenous leukemia.
In this paper we demonstrate that maturing neoplastic cells from patients with chronic myelogenous leukemia (CML) constitutively produce G-CSF and are also receptive for this molecule. G-CSF functions as an autocrine growth factor in stable phase CML, and thus is responsible for divisions of maturing leukemic cells leading to an expansion of the compartment of mature cells. This observation is well in line with in vivo features of CML in stable phase, i.e., the hyperplasia of the mature granulocyte compartment. In acute blastic phase of CML expression of the G-CSF gene seems to be less common and not related to autonomous blast growth.


Effect of vacuum-condensed or ultrafiltered milk on pasteurized process cheese.
Milk was concentrated by ultrafiltration (UF) or vacuum condensing (CM) and milks with 2 levels of protein: 4.5% (UF1 and CM1) and 6.0% (UF2 and CM2) for concentrates and a control with 3.2% protein were used for manufacturing 6 replicates of Cheddar cheese. For manufacturing pasteurized process cheese, a 1:1 blend of shredded 18- and 30-wk Cheddar cheese, butter oil, and disodium phosphate (3%) was heated and pasteurized at 74 degrees C for 2 min with direct steam injection. The moisture content of the resulting process cheeses was 39.4 (control), 39.3 (UF1), 39.4 (UF2), 39.4 (CM1), and 40.2% (CM2). Fat and protein contents were influenced by level and method of concentration of cheese milk. Fat content was the highest in control (35.0%) and the lowest in UF2 (31.6%), whereas protein content was the lowest in control (19.6%) and the highest in UF2 (22.46%). Ash content increased with increase in level of concentration of cheese milk with no effect of method of concentration. Meltability of process cheeses decreased with increase in level of concentration and was higher in control than in the cheeses made with concentrated milk. Hardness was highest in UF cheeses (8.45 and 9.90 kg for UF1 and UF2) followed by CM cheeses (6.27 and 9.13 kg, for CM1 and CM2) and controls (3.94 kg). Apparent viscosity of molten cheese at 80 degrees C was higher in the 6.0% protein treatments (1043 and 1208 cp, UF2 and CM2) than in 4.5% protein treatments (855 and 867 cp, UF1 and CM1) and in control (557 cp). Free oil in process cheeses was influenced by both level and method of concentration with control (14.3%) being the lowest and CM2 (18.9%) the highest. Overall flavor, body and texture, and acceptability were higher for process cheeses made with the concentrates compared with control. This study demonstrated that the application of concentrated milks (UF or CM) for Cheddar cheese making has an impact on pasteurized process cheese characteristics.


Low energy electron-induced decomposition of (η3-C3H5)Ru(CO)3Br, a potential focused electron beam induced deposition precursor with a heteroleptic ligand set.
Here we describe in detail low energy electron induced fragmentation of a potential focused electron beam induced deposition (FEBID) precursor, π-allyl ruthenium tricarbonyl bromide, i.e. (η3-C3H5)Ru(CO)3Br, specially designed to allow comparison of the effect of different ligands on the efficiency of low energy electron induced fragmentation of FEBID precursors. Specifically, we discuss the efficiency of dissociative electron attachment (DEA) and dissociative ionization (DI) with respect to electron-induced removal of the allyl, bromide and carbonyl ligands. We place this in perspective with a previous surface study on the same precursor and we propose a design strategy for FEBID precursor molecules to increase their susceptibility towards DEA.


Historical land-use and landscape change in southern Sweden and implications for present and future biodiversity.
The two major aims of this study are (1) To test the performance of the Landscape Reconstruction Algorithm (LRA) to quantify past landscape changes using historical maps and related written sources, and (2) to use the LRA and map reconstructions for a better understanding of the origin of landscape diversity and the recent loss of species diversity. Southern Sweden, hemiboreal vegetation zone. The LRA was applied on pollen records from three small bogs for four time windows between AD 1700 and 2010. The LRA estimates of % cover for woodland/forest, grassland, wetland, and cultivated land were compared with those extracted from historical maps within 3-km radius around each bog. Map-extracted land-use categories and pollen-based LRA estimates (in % cover) of the same land-use categories show a reasonable agreement in several cases; when they do not agree, the assumptions used in the data (maps)-model (LRA) comparison are a better explanation of the discrepancies between the two than possible biases of the LRA modeling approach. Both the LRA reconstructions and the historical maps reveal between-site differences in landscape characteristics through time, but they demonstrate comparable, profound transformations of the regional and local landscapes over time and space due to the agrarian reforms in southern Sweden during the 18th and 19th centuries. The LRA was found to be the most reasonable approach so far to reconstruct quantitatively past landscape changes from fossil pollen data. The existing landscape diversity in the region at the beginning of the 18th century had its origin in the long-term regional and local vegetation and land-use history over millennia. Agrarian reforms since the 18th century resulted in a dramatic loss of landscape diversity and evenness in both time and space over the last two centuries leading to a similarly dramatic loss of species (e.g., beetles).


Memory consolidation in aging and MCI after 1 week.
To assess consolidation in amnestic mild cognitive impairment (aMCI), controlling for differences in initial learning and using a protracted delay period for recall. 15 individuals with aMCI were compared with 15 healthy older adult controls on a story learning task. Subjects were trained to criteria to equalize initial learning across subjects. Recall was tested at both the 30-min typically used delay and a 1-week delay used to target consolidation. Using repeated measures ANOVAs adjusted for age, we found group × time point interactions across the entire task between the final trial and 30-min delay, and again between the 30-min and 1-week delay periods, with aMCI having greater declines in recall as compared with controls. Significant group main effects were also found, with aMCI recalling less than controls. Consolidation was impaired in aMCI as compared with controls. Our findings indicate that aMCI-related performance typically measured at 30 min underestimates aMCI-associated memory deficits. This is the first study to isolate consolidation by controlling for initial learning differences and using a protracted delay period to target consolidation in an aMCI sample.


HIV-1 reverse transcriptase-associated RNase H cleaves RNA/RNA in arrested complexes: implications for the mechanism by which RNase H discriminates between RNA/RNA and RNA/DNA.
Reverse transcription of human immunodeficiency virus type 1 (HIV-1) is primed by tRNA(Lys3), which forms an 18 base pair RNA homoduplex with its 3' terminus and the primer binding site (PBS) of the viral genome. Using an in vitro system mimicking initiation of minus strand DNA synthesis, we analyzed the mechanism by which HIV-1 reverse transcriptase (RT)-associated ribonuclease H (RNase H) distinguishes between RNA/DNA and RNA/RNA (dsRNA). tRNA(Lys3) was hybridized to a PBS-containing RNA template and extended by addition of deoxynucleoside triphosphates (dNTPs). In the presence of all four dNTPs, initial cleavage of the RNA template occurred immediately downstream of the tRNA-DNA junction, reflecting RNase H specificity for RNA in a RNA/DNA hybrid. However, in the absence of DNA synthesis, or limiting this by chain termination, the PBS was cleaved at a constant distance of 18 nucleotides upstream of the nascent primer 3' terminus. The position of cleavage remained in register with the position of DNA synthesis arrest, indicating that hydrolysis of homoduplex RNA is spatialy co-ordinated with DNA synthesis. Kinetic studies comparing cleavage rates of an analogous DNA primer/PBS heteroduplex and the tRNA(Lys3)/PBS homoduplex showed that while the former is cleaved as rapidly as RT polymerizes, the latter proceeds 30-fold slower. Although the RNase H domain hydrolyzes dsRNA when RT is artificially arrested, specificity for RNA/DNA hybrids is maintained when DNA is actively synthesized, since residency of the RNase H domain at a single base position is not long enough to allow significant cleavage on dsRNA.


Sleep enhances learning of a functional motor task in young adults.
Sleep has been demonstrated to enhance simple motor skill learning "offline" in young adults. "Offline learning" refers to either the stabilization or the enhancement of a memory through the passage of time without additional practice. It remains unclear whether a functional motor task will benefit from sleep to produce offline motor skill enhancement. Physical therapists often teach clients functional motor skills; therefore, it is important to understand how sleep affects learning of these skills. The purpose of this study was to determine whether sleep enhances the learning of a functional motor task. A prospective, cross-sectional, repeated-measures design was used. Young participants who were healthy (N=24) were randomly assigned to either a sleep group or a no-sleep group. The sleep group practiced a novel walking task in the evening and underwent retention testing the following morning, and the no-sleep group practiced the task in the morning and underwent retention testing in the evening. Outcome measures included time around the walking path and spatiotemporal gait parameters. Only participants who slept after practicing the novel walking task demonstrated a significant offline improvement in performance. Compared with the no-sleep group, participants in the sleep group demonstrated a significant decrease in the time around the walking path, an increase in tandem velocity, an increase in tandem step length, and a decline in tandem step time. Time-of-day effect and inability to ensure a certain amount of sleep quantity and quality of participants were limitations of the study. This study is the first to provide evidence that sleep facilitates learning clinically relevant functional motor tasks. Sleep is an important factor that physical therapists should consider when teaching clients motor skills.


Lack of clonal relationship between non-typhi Salmonella strain types from humans and those isolated from animals living in close contact.
Antibiogram patterns and chromosomal DNA typing were used to compare 151 non-typhoidal Salmonella spp. (NTS) isolated from patients and 78 from animals, environmental or food specimens obtained within or near the homes of patients with invasive salmonellosis. The majority of NTS from humans (137; 90.7%) were Salmonella enterica serotype Typhimurium (S. Typhimurium) and S. Enteritidis. Chicken specimens and feeds produced (24; 52.2%) S. Enteritidis, while S. Agona was the predominant (20; 77%) serovar among pigs and dairy cows. The majority (97; 64.2%) of NTS from humans were multidrug resistant, while NTS from cows, pigs, beef carcass swabs and sewers were fully susceptible to all antibiotics tested. Pulsed-field gel electrophoresis patterns of XbaI-digested genomic DNA of NTS from the humans and the chickens were different. However, S. Enteritidis from chickens, and S. Braenderup and S. Agona from cows and pigs were clustered together in one group. There was no significant relatedness between NTS isolates from humans and those from animals, food or the environment in close contact to humans.


Fibronectin regulates calvarial osteoblast differentiation.
The secretion of fibronectin by differentiating osteoblasts and its accumulation at sites of osteogenesis suggest that fibronectin participates in bone formation. To test this directly, we determined whether fibronectin-cell interactions regulate progressive differentiation of cultured fetal rat calvarial osteoblasts. Spatial distributions of alpha 5 integrin subunit, fibronectin, osteopontin (bone sialoprotein I) and osteocalcin (bone Gla-protein) were similar in fetal rat calvaria and mineralized, bone-like nodules formed by cultured osteoblasts. Addition of anti-fibronectin antibodies to cultures at confluence reduced subsequent formation of nodules to less than 10% of control values, showing that fibronectin is required for normal nodule morphogenesis. Anti-fibronectin antibodies selectively inhibited steady-state expression of mRNA for genes associated with osteoblast differentiation; mRNA levels for alkaline phosphatase and osteocalcin were suppressed, whereas fibronectin, type I collagen and osteopontin were unaffected. To identify functionally relevant domains of fibronectin, we treated cells with soluble fibronectin fragments and peptides. Cell-binding fibronectin fragments (type III repeats 6-10) containing the Arg-Gly-Asp (RGD) sequence blocked both nodule initiation and maturation, whether or not they contained a functional synergy site. In contrast, addition of the RGD-containing peptide GRGDSPK alone did not inhibit nodule initiation, although it did block nodule maturation. Thus, in addition to the RGD sequence, other features of the large cell-binding fragments contribute to the full osteogenic effects of fibronectin. Nodule formation and osteoblast differentiation resumed after anti-fibronectin antibodies or GRGDSPK peptides were omitted from the media, showing that the inhibition was reversible and the treatments were not cytotoxic. Outside the central cell-binding domain, peptides from the IIICS region and antibodies to the N terminus did not inhibit nodule formation. We conclude that osteoblasts interact with the central cell-binding domain of endogenously produced fibronectin during early stages of differentiation, and that these interactions regulate both normal morphogenesis and gene expression.


KOTOBUKI-1 apparatus for cryogenic coherent X-ray diffraction imaging.
We have developed an experimental apparatus named KOTOBUKI-1 for use in coherent X-ray diffraction imaging experiments of frozen-hydrated non-crystalline particles at cryogenic temperature. For cryogenic specimen stage with small positional fluctuation for a long exposure time of more than several minutes, we here use a cryogenic pot cooled by the evaporation cooling effect for liquid nitrogen. In addition, a loading device is developed to bring specimens stored in liquid nitrogen to the specimen stage in vacuum. The apparatus allows diffraction data collection for frozen-hydrated specimens at 66 K with a positional fluctuation of less than 0.4 μm and provides an experimental environment to easily exchange specimens from liquid nitrogen storage to the specimen stage. The apparatus was developed and utilized in diffraction data collection of non-crystalline particles with dimensions of μm from material and biological sciences, such as metal colloid particles and chloroplast, at BL29XU of SPring-8. Recently, it has been applied for single-shot diffraction data collection of non-crystalline particles with dimensions of sub-μm using X-ray free electron laser at BL3 of SACLA.


Rabring7 degrades c-Myc through complex formation with MM-1.
We have reported that a novel c-Myc-binding protein, MM-1, repressed E-box-dependent transcription and transforming activities of c-Myc and that a mutation of A157R in MM-1, which is often observed in patients with leukemia or lymphoma, abrogated all of the repressive activities of MM-1 toward c-Myc, indicating that MM-1 is a novel tumor suppressor. MM-1 also binds to the ubiquitin-proteasome system, leading to degradation of c-Myc. In this study, we identified Rabring7, a Rab7-binding and RING finger-containing protein, as an MM-1-binding protein, and we found that Rabring7 mono-ubiquitinated MM-1 in the cytoplasm without degradation of MM-1. Rabring7 was also found to bind to c-Myc and to ubiquitinate c-Myc in a threonine 58-dependent manner. When c-Myc was co-transfected with MM-1 and Rabring7, c-Myc was degraded. Furthermore, it was found that c-Myc was stabilized in MM-1-knockdown cells even when Rabring7 was transfected and that Rabring7 was bound to and co-localized with MM-1 and c-Myc after MM-1 and Rabring7 had been translocated from the cytoplasm to the nucleus. These results suggest that Rabring7 stimulates c-Myc degradation via mono-ubiquitination of MM-1.


Analgesic antipyretic use among young children in the TEDDY study: no association with islet autoimmunity.
The use of analgesic antipyretics (ANAP) in children have long been a matter of controversy. Data on their practical use on an individual level has, however, been scarce. There are indications of possible effects on glucose homeostasis and immune function related to the use of ANAP. The aim of this study was to analyze patterns of analgesic antipyretic use across the clinical centers of The Environmental Determinants of Diabetes in the Young (TEDDY) prospective cohort study and test if ANAP use was a risk factor for islet autoimmunity. Data were collected for 8542 children in the first 2.5 years of life. Incidence was analyzed using logistic regression with country and first child status as independent variables. Holm's procedure was used to adjust for multiplicity of intercountry comparisons. Time to autoantibody seroconversion was analyzed using a Cox proportional hazards model with cumulative analgesic use as primary time dependent covariate of interest. For each categorization, a generalized estimating equation (GEE) approach was used. Higher prevalence of ANAP use was found in the U.S. (95.7%) and Sweden (94.8%) compared to Finland (78.1%) and Germany (80.2%). First-born children were more commonly given acetaminophen (OR 1.26; 95% CI 1.07, 1.49; p = 0.007) but less commonly Non-Steroidal Anti-inflammatory Drugs (NSAID) (OR 0.86; 95% CI 0.78, 0.95; p = 0.002). Acetaminophen and NSAID use in the absence of fever and infection was more prevalent in the U.S. (40.4%; 26.3% of doses) compared to Sweden, Finland and Germany (p < 0.001). Acetaminophen or NSAID use before age 2.5 years did not predict development of islet autoimmunity by age 6 years (HR 1.02, 95% CI 0.99-1.09; p = 0.27). In a sub-analysis, acetaminophen use in children with fever weakly predicted development of islet autoimmunity by age 3 years (HR 1.05; 95% CI 1.01-1.09; p = 0.024). ANAP use in young children is not a risk factor for seroconversion by age 6 years. Use of ANAP is widespread in young children, and significantly higher in the U.S. compared to other study sites, where use is common also in absence of fever and infection.


The theory of organisational socialisation and its potential for improving transition experiences for new graduate nurses.
Graduate nurse transition continues to remain a difficult time for many new graduate nurses, with significant numbers of graduates being dissatisfied, ultimately considering leaving or exiting the profession. Currently, many graduate nurse programs within Australia and internationally reflect a homogeneous nature pertaining to content and program delivery. A refinement of graduate nurse transition programs through an adaptation of a model of organisational socialisation supports a more individualised approach to transition, improving graduate outcomes and addressing attrition rates. To propose a model which supports the accommodation of new graduates within a health service improving both new graduate and health service outcomes through; greater levels of job satisfaction, increased commitment to an organisation and decreased turnover of new staff. Theoretical paper based on a program of research. An adaptation of a model of organisational socialisation was applied to the process of transition for newly qualified graduate nurses. This adaptation was informed by a larger 2012 Australian study (findings reported extensively elsewhere) with 459 newly qualified graduate nurses reporting their transition experiences of the first year of practice. Newly qualified graduate nurses reported effective socialisation with transition based on the following; enduring and continuous orientation throughout the first year of practice, allocation of patient responsibilities reflecting a level of acuity commensurate with a beginning skill set to meet care needs, and feedback of a respectful nature to improve confidence and competence in practice. Negative transition experiences were noted by many new graduates if these factors were not considered. Graduate nurse turnover is costly and destabilising for health services. One means of addressing this is the creation of positive working environments which appropriately socialise new graduates into health services. Accommodating new employees through; individual recognition, modelling of behaviours and developing positive transition outcomes will improve graduate nurse satisfaction and importantly retention.


Active school transport and weekday physical activity in 9-11-year-old children from 12 countries.
Active school transport (AST) may increase the time that children spend in physical activity (PA). This study examined relationships between AST and weekday moderate-to-vigorous physical activity (MVPA), light physical activity (LPA), sedentary time (SED) and total activity during naturally organized time periods (daily, before school, during school and after school) in a sample of children from 12 countries. The sample included 6224 children aged 9-11 years. PA and sedentary time were objectively measured using Actigraph accelerometers. AST was self-reported by participants. Multilevel generalized linear and logistic regression statistical models were used to determine associations between PA, SED and AST across and within study sites. After adjustment for age, highest parental educational attainment, BMI z-score and accelerometer wear time, children who engaged in AST accumulated significantly more weekday MVPA during all studied time periods and significantly less time in LPA before school compared with children who used motorized transport to school. AST was unrelated to time spent in sedentary behaviors. Across all study sites, AST was associated with 6.0 min (95% confidence interval (CI): 4.7-7.3; P<0.0001) more of weekday MVPA; however, there was some evidence that this differed across study sites (P for interaction=0.06). Significant positive associations were identified within 7 of 12 study sites, with differences ranging from 4.6 min (95% CI: 0.3-8.9; P=0.04, in Canada) to 10.2 min (95% CI: 5.9-14.4; P<0.0001, in Brazil) more of daily MVPA among children who engaged in AST compared with motorized transport. The present study demonstrated that AST was associated with children spending more time engaged in MVPA throughout the day and less time in LPA before school. AST represents a good behavioral target to increase levels of PA in children.


Diverse echocardiographic changes in the course of hypoxia due to acute exacerbation of idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a progressive parenchymal disease. Pulmonary hypertension (PH) is a potentially lethal complication in the course of IPF. In almost all cases of IPF-PH there is gradual deterioration, but patients can also decline suddenly due to hypoxia. This case report describes the different echocardiographic changes observed in 2 episodes of hypoxic attack in a 73-year-old man. On admission, the tricuspid regurgitation peak gradient (TRPG) was 21 mmHg and the oxygen saturation rate was 94% (O2: 4 L/min). Five days after admission, the TRPG and oxygen saturation rate deteriorated [TRPG: 85 mmHg, oxygen saturation: 72% (O2; 4 L/min)]. He was diagnosed with IPF-PH due to hypoxic pulmonary vasoconstriction. Oxygen therapy and methylprednisolone pulse therapy (MPT) were administered. Five days after the MPT treatment, the hypoxia and PH improved [TRPG: 21 mmHg, oxygen saturation: 95% (O2: 4 L/min)]. Acute exacerbation of IPF (IPF-AE) occurred 20 days after the MPT, and a second dose of MPT was administered. The TRPG and oxygen saturation rate did not decline [TRPG: 27 mmHg, oxygen saturation: 94% (O2: 4 L/min)]. The patient died 10 days after the second dose of MPT. Divergent echocardiographic findings were observed during the deterioration of IPF-AE in the presence of IPF-PH.


Bronchopulmonary disease in Iranian housewives chronically exposed to indoor smoke.
Tobacco smoking is recognized as the main cause of chronic bronchitis followed by occupational exposure to dust, fumes and gases. Indoor inhalation of smoke is being investigated increasingly but mainly the acute morbidity has been described while the late features and complications which may lead to respiratory disability are less well known. Among the 388 female patients who were referred to the main Bronchoscopy Unit of Tehran University of Medical Sciences between 1986 and 1995, 10 nonsmoking elderly housewives were found with chronic bronchopulmonary lesions characterized by widespread mucosal swelling and anthracotic plaques of the airways. Enquiry into their past history revealed massive exposure to smoke over many years whilst baking household rustic bread inside dwellings. No other source of air pollution was detected. The main clinicopathological features were respiratory disability due to chronic bronchitis with intracellular as well as extracellular black particles. As similar cases may present to medical practitioners even in affluent societies as a result of increasing amounts of immigrants from developing countries, recognition of these patients is important to avoid irrelevant diagnoses and unnecessary interventions.


Whole Transcriptome Analysis of Pre-invasive and Invasive Early Squamous Lung Carcinoma in Archival Laser Microdissected Samples.
Preinvasive squamous cell cancer (PSCC) are local transformations of bronchial epithelia that are frequently observed in current or former smokers. Their different grades and sizes suggest a continuum of dysplastic change with increasing severity, which may culminate in invasive squamous cell carcinoma (ISCC). As a consequence of the difficulty in isolating cancerous cells from biopsies, the molecular pathology that underlies their histological variability remains largely unknown. To address this issue, we have employed microdissection to isolate normal bronchial epithelia and cancerous cells from low- and high-grade PSCC and ISCC, from paraffin embedded (FFPE) biopsies and determined gene expression using Affymetric Human Exon 1.0 ST arrays. Tests for differential gene expression were performed using the Bioconductor package limma followed by functional analyses of differentially expressed genes in IPA. Examination of differential gene expression showed small differences between low- and high-grade PSCC but substantial changes between PSCC and ISCC samples (184 vs 1200 p-value <0.05, fc ±1.75). However, the majority of the differentially expressed PSCC genes (142 genes: 77%) were shared with those in ISCC samples. Pathway analysis showed that these shared genes are associated with DNA damage response, DNA/RNA metabolism and inflammation as major biological themes. Cluster analysis identified 12 distinct patterns of gene expression including progressive up or down-regulation across PSCC and ISCC. Pathway analysis of incrementally up-regulated genes revealed again significant enrichment of terms related to DNA damage response, DNA/RNA metabolism, inflammation, survival and proliferation. Altered expression of selected genes was confirmed using RT-PCR, as well as immunohistochemistry in an independent set of 45 ISCCs. Gene expression profiles in PSCC and ISCC differ greatly in terms of numbers of genes with altered transcriptional activity. However, altered gene expression in PSCC affects canonical pathways and cellular and biological processes, such as inflammation and DNA damage response, which are highly consistent with hallmarks of cancer.


Mortality and pathology in brown bullheads Amieurus nebulosus associated with a spontaneous Edwardsiella ictaluri outbreak under tank culture conditions.
Brown bullheads Amieurus nebulosus (family Ictaluridae) are commonly used as a sentinel of environmental contamination. These fish are not generally cultured under laboratory conditions and little is known about their disease susceptibility. Here we report an outbreak of disease due to Edwardsiella ictaluri in a laboratory population of tank-reared, wild-caught brown bullheads. The isolate was positively identified as E. ictaluri using standard bacteriological substrate utilization tests and a monoclonal antibody specific for this bacterium. This pathogen causes a significant disease in channel catfish Ictalurus punctatus and is associated with disease in other ictalurid and non-ictalurid fishes. It appears that E. ictaluri is also a significant pathogen in brown bullheads and produces clinical signs and lesions similar but not identical to those observed in channel catfish. Since commercial sources of bullheads for laboratory tank studies are not available, precautions should be taken to prevent potential E. ictaluri disease outbreaks from wild-caught bullheads intended for laboratory research.


The detailed acoustic signature of a micro-confined cavitation bubble.
Numerous scenarios exist for a cavitation bubble growing in a liquid. We focus here on cavitation phenomena within water under static tension in a confined environment. Drawing inspiration from the natural materials in plants, we design a novel experimental setup where a micrometric volume of water is confined by a hydrogel-based material. We show that, submerging the sample in a hypertonic solution, the water within the cavity is placed under tension and the acoustic emission produced by the resulting bubble nucleation is precisely detected. This new experimental procedure is able to strongly reduce the acoustic reflections occurring at the hydrogel/air interface with more classical techniques. We also propose a mathematical model to characterise the pressure wave emitted in order to correctly take into account the dissipation effect induced by the visco-elastic behaviour of the confining hydrogel. Both bubble resonant frequency and damping are captured by the model and quantitatively match the values found in the experiments.


Substitution model of sequence evolution for the human immunodeficiency virus type 1 subtype B gp120 gene over the C2-V5 region.
Phylogenetic analyses frequently rely on models of sequence evolution that detail nucleotide substitution rates, nucleotide frequencies, and site-to-site rate heterogeneity. These models can influence hypothesis testing and can affect the accuracy of phylogenetic inferences. Maximum likelihood methods of simultaneously constructing phylogenetic tree topologies and estimating model parameters are computationally intensive, and are not feasible for sample sizes of 25 or greater using personal computers. Techniques that initially construct a tree topology and then use this non-maximized topology to estimate ML substitution rates, however, can quickly arrive at a model of sequence evolution. The accuracy of this two-step estimation technique was tested using simulated data sets with known model parameters. The results showed that for a star-like topology, as is often seen in human immunodeficiency virus type 1 (HIV-1) subtype B sequences, a random starting topology could produce nucleotide substitution rates that were not statistically different than the true rates. Samples were isolated from 100 HIV-1 subtype B infected individuals from the United States and a 620 nt region of the env gene was sequenced for each sample. The sequence data were used to obtain a substitution model of sequence evolution specific for HIV-1 subtype B env by estimating nucleotide substitution rates and the site-to-site heterogeneity in 100 individuals from the United States. The method of estimating the model should provide users of large data sets with a way to quickly compute a model of sequence evolution, while the nucleotide substitution model we identified should prove useful in the phylogenetic analysis of HIV-1 subtype B env sequences.


In vitro and in vivo evaluation of PEDOT microelectrodes for neural stimulation and recording.
Cortical neural prostheses require chronically implanted small-area microelectrode arrays that simultaneously record and stimulate neural activity. It is necessary to develop new materials with low interface impedance and large charge transfer capacity for this application and we explore the use of conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) for the same. We subjected PEDOT coated electrodes to voltage cycling between -0.6 and 0.8 V, 24 h continuous biphasic stimulation at 3 mC/cm² and accelerated aging for four weeks. Characterization was performed using cyclic voltammetry, electrochemical impedance spectroscopy, and voltage transient measurements. We found that PEDOT coated electrodes showed a charge injection limit 15 times higher than Platinum Iridium (PtIr) electrodes and electroplated Iridium Oxide (IrOx) electrodes when using constant current stimulation at zero voltage bias. In vivo chronic testing of microelectrode arrays implanted in rat cortex revealed that PEDOT coated electrodes show higher signal-to-noise recordings and superior charge injection compared to PtIr electrodes.


Cardiovascular Gene Therapy: Past, Present, and Future.
Cardiovascular diseases remain a large global health problem. Although several conventional small-molecule treatments are available for common cardiovascular problems, gene therapy is a potential treatment option for acquired and inherited cardiovascular diseases that remain with unmet clinical needs. Among potential targets for gene therapy are severe cardiac and peripheral ischemia, heart failure, vein graft failure, and some forms of dyslipidemias. The first approved gene therapy in the Western world was indicated for lipoprotein lipase deficiency, which causes high plasma triglyceride levels. With improved gene delivery methods and more efficient vectors, together with interventional transgene strategies aligned for a better understanding of the pathophysiology of these diseases, new approaches are currently tested for safety and efficacy in clinical trials. In this article, we integrate a historical perspective with recent advances that will likely affect clinical development in this research area.


Properties of neural crest-like cells differentiated from human embryonic stem cells.
Neural crest cells (NCCs) derive early in vertebrate ontogenesis from neural tube as a population of migratory cells with exquisite differentiation potential. Abnormalities in NCC behaviour are cause of debilitating diseases including cancers and a spectrum of neurocristopathies. Thanks to their multilineage differentiation capacity NCCs offer a cell source for regenerative medicine. Both these aspects make NCC biology an important issue to study, which can currently be addressed using methodologies based on pluripotent stem cells. Here we contributed to understanding the biology of human NCCs by refining the protocol for differentiation/propagation of NCClike cells from human embryonic stem cells and by characterizing the molecular and functional phenotype of such cells. Most importantly, we improved formulation of media for NCC culture, we found that poly-L-ornithine combined with fibronectin provide good support for NCC growth, we unravelled the tendency of cultured NCCs to maintain heterogeneity of CD271 expression, and we showed that NCCs derived here possess the capacity to react to BMP4 signals by dramatically up-regulating MSX1, which is linked to odontogenesis.


Magnesium Is a Key Player in Neuronal Maturation and Neuropathology.
Magnesium (Mg) is the second most abundant cation in mammalian cells, and it is essential for numerous cellular processes including enzymatic reactions, ion channel functions, metabolic cycles, cellular signaling, and DNA/RNA stabilities. Because of the versatile and universal nature of Mg2+, the homeostasis of intracellular Mg2+ is physiologically linked to growth, proliferation, differentiation, energy metabolism, and death of cells. On the cellular and tissue levels, maintaining Mg2+ within optimal levels according to the biological context, such as cell types, developmental stages, extracellular environments, and pathophysiological conditions, is crucial for development, normal functions, and diseases. Hence, Mg2+ is pathologically involved in cancers, diabetes, and neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease, and demyelination. In the research field regarding the roles and mechanisms of Mg2+ regulation, numerous controversies caused by its versatility and complexity still exist. As Mg2+, at least, plays critical roles in neuronal development, healthy normal functions, and diseases, appropriate Mg2+ supplementation exhibits neurotrophic effects in a majority of cases. Hence, the control of Mg2+ homeostasis can be a candidate for therapeutic targets in neuronal diseases. In this review, recent results regarding the roles of intracellular Mg2+ and its regulatory system in determining the cell phenotype, fate, and diseases in the nervous system are summarized, and an overview of the comprehensive roles of Mg2+ is provided.


SMAD3 represses androgen receptor-mediated transcription.
The androgen-signaling pathway is important in the growth and progression of prostate cancer. Androgen ablation therapy, which may result in programmed cell death, is often used to treat advanced prostate cancer. The growth-promoting effects of androgen are mediated mostly through the androgen receptor (AR). Transforming growth factor beta (TGF-beta) plays critical roles in controlling prostate cell proliferation, differentiation, and apoptosis. Normal transcripts and proteins of TGF-beta receptors are frequently lost in prostate cancer cells, especially in advanced stages of the disease. However, the mechanisms by which TGF-beta inhibits proliferation and induces apoptosis in prostate cancer cells is not clear. We investigated the molecular mechanism by which TGF-beta inhibits transcriptional activation mediated by AR. Using transient transfection systems, we demonstrated that Smad3 specifically represses transcriptional activation mediated by AR on two natural androgen-responsive promoters. This repression is transmitted through TGF-beta signaling and can be regulated by other Smad proteins. A protein-protein interaction between AR and Smad3 was identified in vitro and in vivo, and the transcription activation domain of AR and the MH2 of Smad3 were identified as being responsible for binding. Additional functional experiments showed that the repression of AR by Smad3 is mediated solely through the MH2 domain. These results provide fresh insight for understanding the mechanism by which TGF-beta regulates the androgen-signaling pathway in prostate cancer cells.


The competitive effects of health maintenance organizations: another look at the evidence from Hawaii, Rochester, and Minneapolis/St. Paul.
There has been much discussion about the potential cost-containing impact of HMOs upon the local medical care market. Three areas have been identified by various observers as experiencing such beneficial effects: Hawaii, after the development of Kaiser in the late 1950s; Rochester, New York, which experienced rapid HMO growth and declining Blue Cross hospital use in the late 1970s; and Minneapolis/St. Paul, which has been the focus of vigorous HMO competition in the last decade. While comprehensive data on health care expenditures are not available, bits of evidence can be pieced together to develop case studies of each area. Careful review of the available data often identifies internal inconsistencies and contradictions, but in none of the three sites is there a reduction in hospital use that is most plausibly attributed to HMO competition. Instead, the reported reductions are in each case attributable to other factors--including biases in data, long-term trends predating HMOs, indirect effects of other policy changes, and other forms of competition.


Effect of panretinal photocoagulation on corneal sensation and the corneal subbasal nerve plexus in diabetes mellitus.
To assess the effects of panretinal photocoagulation (PRP) for diabetic retinopathy (DR) on the human corneal subbasal nerve plexus (SBNP) and to investigate correlations between corneal subbasal nerve (SBN) density, corneal sensitivity, and diabetic peripheral neuropathy. Thirty-eight subjects with at least a 10-year history of diabetes mellitus (DM) or DR were included. Subjects were assigned to a PRP group (n = 19), having undergone a treatment of retinopathy in at least one eye or a non-PRP group (n = 19), with no history of PRP. The Michigan Neuropathy Screening Instrument (MNSI) was administered to enable quantification of neuropathic symptoms. Laser scanning in vivo confocal microscopy was performed to capture images of the corneal SBNP to allow determination of SBNP density. Central corneal sensitivity (CST) was evaluated by noncontact aesthesiometry and peripheral vibration perception threshold was measured with a biothesiometer. Mean SBNP densities were 12.27 ± 4.28 mm/mm²) in the PRP group and 12.75 ± 3.59 mm/mm² in the non-PRP group. There were no significant differences in SBNP density (P = 0.71), CST (P = 0.84), MNSI score (P = 0.19), and biothesiometry (P = 0.77) between the PRP and non-PRP groups. When data from both groups (n = 38) were combined, corneal sensitivity was modestly correlated with SBNP density (r = 0.30, P = 0.06), and peripheral biothesiometry (r = 0.26, P = 0.11). In DM correlation of corneal sensitivity, SBNP density, and peripheral biothesiometry may have a potential role in estimating the severity of peripheral neuropathy. Corneal SBNP density and sensitivity appear to be unaffected by PRP laser treatment compared with non-PRP diabetic eyes.


Whole blood chloroquine concentrations with Plasmodium vivax infection in Irian Jaya, Indonesia.
Whole blood concentrations of self-administered chloroquine (CQ) and its metabolite desethylchloroquine (DCQ) were measured in 168 patients with microscopically confirmed infection by Plasmodium vivax in northeastern Irian Jaya, Indonesia. The study consisted of both survey and passive case detection in four separate villages between 1992 and 1994. The subjects were Javanese people 4-51 years old who had lived in the Arso region for up to two years. The sum of CQ and DCQ ranged from 0 to 8,342 ng/ml of whole blood, and 122 subjects (73%) had > or = 100 ng/ml of CQ plus DCQ, the estimated minimally effective concentration (MEC) in whole blood against chloroquine-sensitive P. vivax. Among 56 subjects reporting to a clinic with symptoms of malaria, 53 (95%) had ordinarily effective levels of chloroquine in blood. Among 109 largely asymptomatic malaria patients found by survey case detection, 69 (63%) had chloroquine blood levels greater than the MEC. Virtually all clinical and most subclinical vivax malaria in this region occurs despite ordinarily effective levels of chloroquine in blood.


Assessing the effects of anti-homosexuality legislation in Uganda on HIV prevention, treatment, and care services.
Uganda's response to the HIV epidemic has been lauded for its robustness and achievements. However, a key component of HIV prevention programming has been missing, for men who have sex with men (MSM). The main reason cited has been criminalization of male homosexual behavior. In 2009, the Anti-Homosexuality Bill (AHB) was introduced in the parliament to enhance existing anti-homosexuality law. A multi-disciplinary team made a Health Impact Assessment of the proposed AHB. The bill as tabled would severely increase punishments, increased closeting. Social capital of MSM would be eroded by clauses mandating reporting by friends, relatives, and acquaintances. Health-care professionals would have to inform on homosexuals. Mandatory HIV testing would be a blow to programming. Probable disclosure of HIV status in a public space (court) would also be a deterrent. Heftier punishments for those testing positive increases stigma and hobbles subsequent care. The AHB argues for exclusion, and more discrimination targeting persons living with HIV and sexual minorities. It will exacerbate the negative public health consequences of the existing legislation. The government of Uganda should review guidance documents published by authoritative bodies including the World Bank, World Health Organization to develop and bring to scale Human rights-affirming HIV prevention, treatment, and care responses.


Treatment of unresectable hepatoblastoma with liver transplantation in the pediatric population.
The purpose of our study was to evaluate the outcome of children who underwent liver transplantation as treatment for unresectable hepatoblastoma. We prospectively collected data on 311 consecutive liver transplants performed at Children's Medical Center of Dallas between October 1984 and November 2000. There were nine recipients (five boys, four girls) with a diagnosis of unresectable hepatoblastoma. Postoperative survival of those currently alive ranged from 6 months to 16 years (mean 6.4 years, median 7.7 years). All recipients received preoperative chemotherapy: 67% received postoperative chemotherapy. Mean AFP level prior to transplantation was 1 448000 ng/mL. Mean age at diagnosis was 0.81 years. Mean age at transplantation was 1.87 years. Only two patients experienced acute cellular rejection in the postoperative period. There was a total of three deaths and one recurrence. The only instance in which AFP levels did not decrease to low or undetectable levels post-transplantation was in the patient with recurrent tumor. Liver transplantation has an established role in the treatment of hepatoblastoma. It accounted for 3% of pediatric liver transplants, and provided the only opportunity for survival in otherwise incurable patients. Early diagnosis and treatment were found to be associated with better results. Response to chemotherapy may be an important factor influencing survival. Rising AFP levels after transplantation are associated with recurrence.


Phase I study of every 2- or 3-week dosing of ramucirumab, a human immunoglobulin G1 monoclonal antibody targeting the vascular endothelial growth factor receptor-2 in patients with advanced solid tumors.
Ramucirumab is a fully human immunoglobulin G1 monoclonal antibody receptor antagonist designed to block the ligand-binding site of vascular endothelial growth factor receptor-2 (VEGFR-2). An initial phase I study evaluated ramucirumab administered weekly in advanced cancer patients. This phase I study of ramucirumab [administered every 2 or 3 weeks (Q2W or Q3W)] examined safety, maximum tolerated dose, pharmacokinetics, immunogenicity, antitumor activity, and pharmacodynamics. Patients with advanced solid malignancies were treated with escalating doses of ramucirumab i.v. over 1 h. Blood was sampled for pharmacokinetics studies throughout treatment; levels of circulating vascular endothelial growth factor-A (VEGF-A) and soluble VEGF receptors (R)-1 and -2 were assessed. Twenty-five patients were treated with ramucirumab: 13 with 6, 8, or 10 mg/kg Q2W, and 12 with 15 or 20 mg/kg Q3W. The median treatment duration was 12 weeks (range 2-81). No dose-limiting toxicities were observed. The most frequently reported adverse events (AEs) included proteinuria and hypertension (n = 6 each), and diarrhea, fatigue and headache (n = 4 each). Treatment-related grade 3/4 AEs were: two grade 3 hypertension (10 and 20 mg/kg), one each grade 3 vomiting, fatigue (20 mg/kg), atrial flutter (15 mg/kg), and one each grade 4 duodenal ulcer hemorrhage (6 mg/kg) and grade 4 pneumothorax (20 mg/kg). Pharmacokinetic analysis revealed low clearance and half-life of ∼110-160 h. Analysis of serum biomarkers indicated considerable patient-to-patient variability, but trends toward elevated VEGF-A and a transient decline in soluble VEGFR-2. Fifteen patients (60%) had best response of stable disease, with a median duration of 13 months (range 2-18 months) in tumor types including colorectal, renal, liver, and neuroendocrine cancers. Ramucirumab was well tolerated. Study results led to recommended phase II doses of 8 mg/kg Q2W and 10 mg/kg Q3W. Prolonged stable disease was observed, suggesting ramucirumab efficacy in various solid tumors. NCT00786383.


Genome-wide association study of 23,500 individuals identifies 7 loci associated with brain ventricular volume.
The volume of the lateral ventricles (LV) increases with age and their abnormal enlargement is a key feature of several neurological and psychiatric diseases. Although lateral ventricular volume is heritable, a comprehensive investigation of its genetic determinants is lacking. In this meta-analysis of genome-wide association studies of 23,533 healthy middle-aged to elderly individuals from 26 population-based cohorts, we identify 7 genetic loci associated with LV volume. These loci map to chromosomes 3q28, 7p22.3, 10p12.31, 11q23.1, 12q23.3, 16q24.2, and 22q13.1 and implicate pathways related to tau pathology, S1P signaling, and cytoskeleton organization. We also report a significant genetic overlap between the thalamus and LV volumes (ρgenetic = -0.59, p-value = 3.14 × 10-6), suggesting that these brain structures may share a common biology. These genetic associations of LV volume provide insights into brain morphology.


moz regulates Hox expression and pharyngeal segmental identity in zebrafish.
In vertebrate embryos, streams of cranial neural crest (CNC) cells migrate to form segmental pharyngeal arches and differentiate into segment-specific parts of the facial skeleton. To identify genes involved in specifying segmental identity in the vertebrate head, we screened for mutations affecting cartilage patterning in the zebrafish larval pharynx. We present the positional cloning and initial phenotypic characterization of a homeotic locus discovered in this screen. We show that a zebrafish ortholog of the human oncogenic histone acetyltransferase MOZ (monocytic leukemia zinc finger) is required for specifying segmental identity in the second through fourth pharyngeal arches. In moz mutant zebrafish, the second pharyngeal arch is dramatically transformed into a mirror-image duplicated jaw. This phenotype resembles a similar but stronger transformation than that seen in hox2 morpholino oligo (hox2-MO) injected animals. In addition, mild anterior homeotic transformations are seen in the third and fourth pharyngeal arches of moz mutants. moz is required for maintenance of most hox1-4 expression domains and this requirement probably at least partially accounts for the moz mutant homeotic phenotypes. Homeosis and defective Hox gene expression in moz mutants is rescued by inhibiting histone deacetylase activity with Trichostatin A. Although we find early patterning of the moz mutant hindbrain to be normal, we find a late defect in facial motoneuron migration in moz mutants. Pharyngeal musculature is transformed late, but not early, in moz mutants. We detect relatively minor defects in arch epithelia of moz mutants. Vital labeling of arch development reveals no detectable changes in CNC generation in moz mutants, but later prechondrogenic condensations are mispositioned and misshapen. Mirror-image hox2-dependent gene expression changes in postmigratory CNC prefigure the homeotic phenotype in moz mutants. Early second arch ventral expression of goosecoid (gsc) in moz mutants and in animals injected with hox2-MOs shifts from lateral to medial, mirroring the first arch pattern. bapx1, which is normally expressed in first arch postmigratory CNC prefiguring the jaw joint, is ectopically expressed in second arch CNC of moz mutants and hox2-MO injected animals. Reduction of bapx1 function in wild types causes loss of the jaw joint. Reduction of bapx1 function in moz mutants causes loss of both first and second arch joints, providing functional genetic evidence that bapx1 contributes to the moz-deficient homeotic pattern. Together, our results reveal an essential embryonic role and a crucial histone acetyltransferase activity for Moz in regulating Hox expression and segmental identity, and provide two early targets, bapx1 and gsc, of moz and hox2 signaling in the second pharyngeal arch.


Merkel cell carcinoma arising from the subcutaneous fat of the arm with intact skin.
Merkel cell carcinoma is a rare malignant neuroendocrine neoplasm characteristically arising from the dermis of sunlight-exposed skin. It rarely arises outside the skin. We present a patient with primary Merkel cell carcinoma arising from subcutaneous fat, with no involvement of the overlying skin. We describe the clinical manifestations and magnetic resonance imaging (MRI) findings. We report a 63-year-old woman with a primary lesion of Merkel cell carcinoma that arose from the subcutaneous fat layer of the left arm. The lesion presented as a subcutaneous nodule with intact overlying skin. MRI showed that the nodular lesion was located entirely in the subcutaneous fat layer, with no involvement of the dermis. Peritumoral infiltration around the lesion and enlarged lymph nodes deep to the lesion were noted. The patient received wide excision of the lesion with dissection of the regional lymph nodes and adjuvant radiotherapy and chemotherapy. Histopathologic examination confirmed the diagnosis of Merkel cell carcinoma with local lymphatic metastasis, and the lesion was completely located in the subcutaneous fat, with no involvement of the dermis. These findings were well correlated with MRI findings. Primary Merkel cell carcinoma may arise from the subcutaneous fat and present as an entirely subcutaneous lesion with intact skin. MRI is helpful to evaluate the local extension of the lesion and regional lymphatic metastasis.


Lipid profile variations in a group of healthy elderly and centenarians.
Epidemiological and clinical studies have clearly shown a close relationship between plasma cholesterol concentrations and vascular risk. We focused our attention on the phenotypic-biohumoral conditions capable of influencing longevity in relation to different age classes. We evaluated the lipid profile in an elderly institutionalized population of 80 subjects (20 males and 60 females divided into age classes) in the town of Catania. Our results revealed a statistically significant reduction in total cholesterol, triglycerides and LDL-cholesterol concentrations as well as Apolipoprotein B100/Apolipoprotein A1, total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL-cholesterol ratios, and a significant increase in HDL-cholesterol, Apolipoprotein A1, Apolipoprotein B100 and Lipoprotein (a) values. This changes are progressive with age. We believe that low total cholesterol, LDL-cholesterol and triglyceride concentrations, elevated HDL-cholesterol values, and low ratios protect subjects from ischemic and thrombotic events, thus favouring longevity. These changes are most evident and statistically significant in the most advanced decades of life, especially in centenarians, and may depend on diverse determinants, such as body composition, environmental factors, physical activity, diet and drugs.


Selective reporting bias of harm outcomes within studies: findings from a cohort of systematic reviews.
To determine the extent and nature of selective non-reporting of harm outcomes in clinical studies that were eligible for inclusion in a cohort of systematic reviews. Cohort study of systematic reviews from two databases. Outcome reporting bias in trials for harm outcomes (ORBIT II) in systematic reviews from the Cochrane Library and a separate cohort of systematic reviews of adverse events. 92 systematic reviews of randomised controlled trials and non-randomised studies published in the Cochrane Library between issue 9, 2012 and issue 2, 2013 (Cochrane cohort) and 230 systematic reviews published between 1 January 2007 and 31 December 2011 in other publications, synthesising data on harm outcomes (adverse event cohort). A 13 point classification system for missing outcome data on harm was developed and applied to the studies. 86% (79/92) of reviews in the Cochrane cohort did not include full data from the main harm outcome of interest of each review for all of the eligible studies included within that review; 76% (173/230) for the adverse event cohort. Overall, the single primary harm outcome was inadequately reported in 76% (705/931) of the studies included in the 92 reviews from the Cochrane cohort and not reported in 47% (4159/8837) of the 230 reviews in the adverse event cohort. In a sample of primary studies not reporting on the single primary harm outcome in the review, scrutiny of the study publication revealed that outcome reporting bias was suspected in nearly two thirds (63%, 248/393). The number of reviews suspected of outcome reporting bias as a result of missing or partially reported harm related outcomes from at least one eligible study is high. The declaration of important harms and the quality of the reporting of harm outcomes must be improved in both primary studies and systematic reviews.


Imaging skeletal muscle using second harmonic generation and coherent anti-Stokes Raman scattering microscopy.
We describe experimental results on label free imaging of striated skeletal muscle using second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy. The complementarity of the SHG and CARS data makes it possible to clearly identify the main sarcomere sub-structures such as actin, myosin, acto-myosin, and the intact T-tubular system as it emanates from the sarcolemma. Owing to sub-micron spatial resolution and the high sensitivity of the CARS microscopy technique we were able to resolve individual myofibrils. In addition, key organelles such as mitochondria, cell nuclei and their structural constituents were observed revealing the entire structure of the muscle functional units. There is a noticeable difference in the CARS response of the muscle structure within actin, myosin and t-tubule areas with respect to laser polarization. We attribute this to a preferential alignment of the probed molecular bonds along certain directions. The combined CARS and SHG microscopy approach yields more extensive and complementary information and has a potential to become an indispensable method for live skeletal muscle characterization.


Expression of the Oenococcus oeni trxA gene is induced by hydrogen peroxide and heat shock.
Sequencing of the DNA region located upstream of the alpha-acetolactate synthase and decarboxylase (alsS-alsD) cluster of Oenococcus oeni allowed identification of an ORF, named trxA. This encodes a protein of 104 amino acids very similar to known thioredoxins. The protein encoded by the cloned fragment was able to complement Escherichia coli strains lacking a functional thioredoxin. Considering the results of protein sequence comparisons and complementation experiments, it was concluded that the trxA gene encodes a functional thioredoxin. Studies of trxA expression showed that the abundance of trxA mRNA was similar during all growth stages. A significant increase in trxA mRNA levels was observed in the presence of hydrogen peroxide in the medium or after heat shock. A single transcriptional start site was determined with total RNA isolated from cells subjected or not subjected to oxidative stress or heat shock. In each case the same promoter region was identified and shown to have a high similarity to the consensus promoter sequence of Gram-positive bacteria, as well as to that of E. coli and the previously mapped promoters from O. oeni.


Pyrolysis of attapulgite clay blended with yak dung enhances pasture growth and soil health: Characterization and initial field trials.
Recent studies have shown that the pyrolysis of biomass combined with clay can result in both lower cost and increase in plant yields. One of the major sources of nutrients for pasture growth, as well as fuel and building materials in Tibet is yak dung. This paper reports on the initial field testing in a pasture setting in Tibet using yak dung, biochar, and attapulgite clay/yak dung biochars produced at ratios of 10/90 and 50/50 clay to dung. We found that the treatment with attapulgite clay/yak dung (50/50) biochar resulted in the highest pasture yields and grass nutrition quality. We also measured the properties and yields of mixtures of clay/yak dung biochar used in the field trials produced at 400°C and 500°C to help determine a possible optimum final pyrolysis temperature and dung/clay ratio. It was observed that increasing clay content increased carbon stability, overall biochar yield, pore size, carboxyl and ketone/aldehyde functional groups, hematite and ferrous/ferric sulphate/thiosulphate concentration, surface area and magnetic moment. Decreasing clay content resulted in higher pH, CEC, N content and an enhanced ability to accept and donate electrons. The resulting properties were a complex function of both processing temperature and the percentage of clay for the biochars processed at both 400°C and 500°C. It is possible that the increase in yield and nutrient uptake in the field trial is related to the higher concentration of C/O functional groups, higher surface area and pore volume and higher content of Fe/O/S nanoparticles of multiple oxidation state in the 50/50 clay/dung. These properties have been found to significantly increase the abundance of beneficial microorganisms and hence improve the nutrient cycling and availability in soil. Further field trials are required to determine the optimum pyrolysis production conditions and application rate on the abundance of beneficial microorganisms, yields and nutrient quality.


Axon plasticity in the mammalian central nervous system after injury.
It is widely recognized that severed axons in the adult central nervous system (CNS) have limited capacity to regenerate. However, mounting evidence from studies of CNS injury response and repair is challenging the prevalent view that the adult mammalian CNS is incapable of structural reorganization to adapt to an altered environment. Animal studies demonstrate the potential to achieve significant anatomical repair and functional recovery following CNS injury by manipulating axon growth regulators alone or in combination with activity-dependent strategies. With a growing understanding of the cellular and molecular mechanisms regulating axon plasticity, and the availability of new experimental tools to map detour circuits of functional importance, directing circuit rewiring to promote functional recovery may be achieved.


GFP-based expression screening of membrane proteins in insect cells using the baculovirus system.
A key step in the production of recombinant membrane proteins for structural studies is the optimization of protein yield and quality. One commonly used approach is to fuse the protein to green fluorescent protein (GFP), enabling expression to be tracked without the need to purify the protein. Combining fusion to green fluorescent protein with the baculovirus expression system provides a useful platform for both screening and production of eukaryotic membrane proteins. In this chapter we describe our protocol for the expression screening of membrane proteins in insect cells using fusion to GFP as a reporter. We use both SDS-PAGE with in-gel fluorescence imaging and fluorescence-detection size-exclusion chromatography (FSEC) to screen for expression.