Light-emitting-diode-induced fluorescence detection of fluorescent dyes for capillary electrophoresis microchip with cross-polarization method.
A cross-polarization scheme is presented to filter out the excitation light from the emission spectrum of fluorescent dyes using green light emitting diodes as a light source and a linear charge coupled device as an intensity detector. The excitation light was linearly polarized and was then used to illuminate the fluorescent dyes in the microchannels of a capillary electrophoresis microchip. The detector was shielded by the second polarizer, oriented perpendicular to the excitation light. The fluorescent signals from Rhodamine B dyes were measured in a dilution series with resulting emission signals and four different concentrations of fluorescent dyes were detected simultaneously with the same excitation source and detector. A limit-of-detection of 1 μM was demonstrated for Rhodamine B dye under the optimal conditions.


Pharmaceutical and pharmacological approaches for bioavailability enhancement of etoposide.
Etoposide, a semi-synthetic derivative of podophyllotoxin, is one of the most active and useful antineoplastic agent used routinely in firstline combination chemotherapy of testicular cancer, small-cell lung cancer and non-Hodgkin's lymphoma. Etoposide displays narrow therapeutic index, erratic pharmacokinetics and dose individualization that needs to be achieved for overcoming inter- and intra-patient variability (25-80 percent), so as to maintain proper drug exposure within a therapeutic range. Etoposide possess high plasma protein binding (97 percent) and is degraded via complex metabolic pathways. The main pharmacokinetic determinants of etoposide are still not completely defined in order to optimize the pharmaco-therapeutic parameters including dose, therapeutic schedule and route of administration. Much research has been done to determine drug-drug and herb-drug interactions for improving the bioavailability of etoposide. The present article gives insight on pharmaceutical and pharmacological attempts made from time to time to overcome the erratic inter- and intra-patient variability for improving the bioavailability of etoposide.


Acute interstitial pneumonia (hamman-rich syndrome) as a cause of idiopathic acute respiratory distress syndrome.
Hamman-Rich syndrome, also known as acute interstitial pneumonia, is a rare and fulminant form of idiopathic interstitial lung disease. It should be considered as a cause of idiopathic acute respiratory distress syndrome. Confirmatory diagnosis requires demonstration of diffuse alveolar damage on lung histopathology. The main treatment is supportive care. It is not clear if glucocorticoid therapy is effective in acute interstitial pneumonia. We report the case of a 77-year-old woman without pre-existing lung disease who initially presented with mild upper respiratory tract infection and then progressed to rapid onset of hypoxic respiratory failure similar to acute respiratory distress syndrome with unknown etiology. Despite glucocorticoid therapy, she did not achieve remission and expired after 35 days of hospitalization. The diagnosis of acute interstitial pneumonia was supported by the histopathologic findings on her lung biopsy.


[Perception of patients with chronic hemiplegia regarding the use of assistive walking devices].
To assess the perception of patients with chronic hemiplegia regarding the use of assistive walking devices (AWD). Twenty-three individuals who met the inclusion criteria and used AWD were recruited from a database of 360 stroke survivors. Their mean age was 58.4 years, mean time since stroke was 80.8 months, and mean time using AWD was 67.6 months. To assess the participants' perception, was used a standardized questionnaire covering the impact of AWDs on five aspects: weight-bearing on the paretic limb; ability to move the paretic limb; confidence; safety; and walking style. Possible answers were "improved," "unchanged," or "decreased." Fourteen individuals used canes and nine used elbow crutches; 21 (91.3%) used AWDs on public roads and only two (8.7%) used AWDs at home. The perception regarding the use of AWDs were positive in the first four questions (6.87 < χ2 < 29.83; 0.0001 < P < 0.03), with reports of improvement in weight-bearing (82.6%), ability to move the paretic limb (39.1%), confidence (82.6%), and safety (86.9%). No significant differences were found in terms of walking style (χ2 = 1.09; P = 0.30). The perception of chronic hemiplegic subjects regarding the use of AWD were positive, suggesting that these devices can improve mobility and independence in activities of daily living.


Computational self-assembly of colloidal crystals from Platonic polyhedral sphere clusters.
We explore a rich phase space of crystals self-assembled from colloidal "polyhedral sphere clusters (PSCs)," each of which consists of equal-sized "halo" spheres placed at the vertices of a polyhedron such that they just touch along each edge. Such clusters, created experimentally by fusing spheres, can facilitate assembly of useful colloidal crystal symmetries not attainable by unclustered spheres. While not crucial for their self-assembly, the center of the PSC can contain a "core" particle that can be used as a scaffold to build the PSC. Using Brownian dynamics simulations, we show the self-assembly of eight distinct crystalline phases from PSCs that correspond to the five Platonic polyhedra, and that are made of spheres with purely repulsive interactions. Strong crystalline order is seen in the centers of mass of the PSCs, or equivalently the core particles. The halo particles also may organize into crystal structures, usually with weaker crystalline order than the core particles. Notably, however, in crystals assembled from the octahedral and icosahedral PSCs, the halo particles are also well ordered, nesting within the crystals formed by the cores. Interestingly, despite the rounded nature of the PSCs, in some cases we obtain structures similar to those of the corresponding faceted polyhedra interacting only via excluded volume. Only the tetrahedral PSCs fail to self-assemble into a crystal, but we demonstrate that a pre-assembled crystal - whose halo particles sit on a close-packed face-centered cubic lattice, and whose core particles form a diamond structure - is stable at high density and melts into a hexagonal phase at lower density.


Possible regeneration of the islets of Langerhans in streptozotocin-diabetic rats given Gymnema sylvestre leaf extracts.
Two water soluble extracts, GS3 and GS4, obtained from the leaves of Gymnema sylvestre, were tested in streptozotocin treated rats for their effects on blood glucose homeostasis and pancreatic endocrine tissue. In the diabetic rats, fasting blood glucose levels returned to normal after 60 days of GS3 and after 20 days of GS4 oral administration. Blood collected during the conduct of oral glucose tolerance tests was used to assay for serum insulin. GS3 and GS4 therapy led to a rise in serum insulin to levels closer to normal fasting levels. In diabetic rat pancreas, both GS3 and GS4 were able to double the islet number and beta cell number. This herbal therapy appears to bring about blood glucose homeostasis through increased serum insulin levels provided by repair/regeneration of the endocrine pancreas.


Transporting neonates with hypoxic-ischemic encephalopathy utilizing active hypothermia.
The objective of this report was to evaluate a servo-controlled active hypothermia device used during the transport of neonates with HIE. Retrospective review of all cases of therapeutic hypothermia, both passive and active, using a servo-controlled device in a single regional referral neonatal intensive care unit from 2009-2013 RESULTS: An ambulance (43%), fixed wing aircraft (25%), or helicopter (32%) transported 28 neonates with active hypothermia. The servo-controlled device captured core temperatures in all 28 neonates, resulting in 2,985 minutes of data. All neonates attained a core temperature between 33-34 °C by 33 minutes of the transport. Once the neonates attained a core temperature, the average temperature for the remainder of the transport was 33 ± 0.2 °C. The neonates maintained the core temperature regardless of the type of transport vehicle. Servo-controlled hypothermia enables rapid attainment of targeted temperatures and maintains the temperature throughout the transport process in both ground and air transport.


Effect of forskolin and isobutylmethylxanthine on delta-opioid receptor activity in neuroblastoma x glioma NG108-15 cells.
Continuous elevation of intracellular cyclic AMP (cAMP) by culturing neuroblastoma x glioma NG108-15 hybrid cells in the presence of forskolin and isobutylmethylxanthine (IBMX) in a chemically defined medium resulted in differentiation of the hybrid cells, as indicated by extension of neurite-like structures and induction of a subclass of G-protein, Go, as monitored by Western analysis. This cellular differentiation also resulted in an initial 25 to 30% increase in [3H]diprenorphine binding 3 hr after forskolin and IBMX treatment, followed by a decrease in opioid receptor density to the maximal level of 35% of control 4 days later. However, the potencies and maximal inhibitory levels of various opioid agonists to inhibit adenylate cyclase activity was not altered during differentiation. When the differentiated hybrid cells were treated with DADLE chronically, an apparent decrease in the ability of the agonist to desensitize and to down-regulate the delta-opioid receptor was observed. It is unlikely that this observed attenuation was due to activation of cAMP-dependent protein kinase A, because (1) attenuation of DADLE desensitization was time-dependent, reaching maximal effects 48 hr after the initiation of treatment; and (2) pretreatment of NG108-15 cells with forskolin and IBMX resulted in attenuation of forskolin's ability to stimulate adenylate cyclase activity and parallel decrease in the ability of forskolin to activate the cAMP-dependent protein kinases in these cells was also observed. Thus, it is unlikely that the activation of protein kinase A by forskolin and IBMX is the cause for the attenuation of DADLE-induced delta-opioid receptor desensitization in differentiated NG108-15 cells.


Controlled Substance Lock-In Programs: Examining An Unintended Consequence Of A Prescription Drug Abuse Policy.
Controlled substance lock-in programs are garnering increased attention from payers and policy makers seeking to combat the epidemic of opioid misuse. These programs require high-risk patients to visit a single prescriber and pharmacy for coverage of controlled substance medication services. Despite high prevalence of the programs in Medicaid, we know little about their effects on patients' behavior and outcomes aside from reducing controlled substance-related claims. Our study was the first rigorous investigation of lock-in programs' effects on out-of-pocket controlled substance prescription fills, which circumvent the programs' restrictions and mitigate their potential public health benefits. We linked claims data and prescription drug monitoring program data for the period 2009-12 for 1,647 enrollees in North Carolina Medicaid's lock-in program and found that enrollment was associated with a roughly fourfold increase in the likelihood and frequency of out-of-pocket controlled substance prescription fills. This finding illuminates weaknesses of lock-in programs and highlights the need for further scrutiny of the appropriate role, optimal design, and potential unintended consequences of the programs as tools to prevent opioid abuse.


Left and right contributions to the Xenopus heart: implications for asymmetric morphogenesis.
The left-right asymmetry of the vertebrate heart is evident in the topology of the heart loop, and in the dissimilar morphology of the left and right chambers. How left-right asymmetric gene expression patterns influence the development of these features is not understood, since the individual roles of the left and right sides of the embryo in heart looping or chamber morphogenesis have not been specifically defined. To this end, we have constructed a bilateral heart-specific fate map of the left and right contributions to the developing heart in the Xenopus embryo. Both the left and right sides contribute to the conoventricular segment of the heart loop; however, the left side contributes to the inner curvature and ventral face of the loop while the right side contributes to the outer curvature and dorsal aspect. In contrast, the left atrium is derived mainly from the original left side of the embryo, while the right atrium is derived primarily from the right side. A comparison of our fate map with the domain of expression of the left-right gene, Pitx2, in the left lateral plate mesoderm, reveals that this Pitx2-expressing region is fated to form the inner curvature of the heart loop, the left atrioventricular canal, and the dorsal aspect of the left atrium. We discuss the implications of these results for the role of left-right asymmetric gene expression in heart looping and chamber morphogenesis.


Survivin-specific CD4+ T cells are decreased in patients with survivin-positive myeloma.
Survivin is a small protein inhibitor of apoptosis and a tumor associated antigen. Survivin expression in multiple myeloma is associated with poor prognosis, disease progression, and drug resistance. The CD4+ response against survivin remains uncharacterized. In order to better understand the anti-tumor immune response to survivin, and optimize vaccination strategies, we characterized the spontaneous CD4+CD25- T cell response against survivin in healthy donors and myeloma patients using survivin derived peptide pools. Healthy donors and myeloma patients' CD4+CD25- T cells exhibited a proliferative and IFN-gamma response against survivin peptides loaded onto autologous dendritic cells. We employed limiting dilution analysis to quantify the precursor frequency of survivin reactive CD4+CD25- T cells. Multiple myeloma patients (range 0% to 2.2x10(-3)%, n=12) had fewer survivin reactive CD4+CD25- T cells than healthy blood donors (range 1.1x10(-3) to 8.4x10(-3)%, n=10), p = 0.021. The survivin reactive CD4+CD25- T cell precursor frequency was inversely associated with tumor survivin mRNA expression (p = 0.0028, r = -1.0, n = 6), and survivin tumor protein expression by IHC (p = 0.0295, r = -0.67, n = 10). A full length mutant survivin protein-pulsed dendritic cell vaccine expanded survivin reactive CD4+CD25- T cells after 12 days of in vitro culture (range 0-540x,median = 42x), and expansion was achieved even in patients with low baseline survivin reactive CD4+ precursors. We have, for the first time, quantified the circulating CD4+CD25- precursor frequency against survivin and demonstrated this is lower in myeloma patients than healthy donors. The number of survivin reactive CD4+CD25- T cells is inversely associated with tumor survivin expression suggesting suppression of survivin responsive CD4+CD25- T cells. Further exploration of a full length mutant survivin protein vaccine which expands survivin reactive CD4+ cells independent of the survivin reactive precursor frequency is warranted.


A holin and an endopeptidase are essential for chitinolytic protein secretion in Serratia marcescens.
Pathogenic bacteria adapt to their environment and manipulate the biochemistry of hosts by secretion of effector molecules. Serratia marcescens is an opportunistic pathogen associated with healthcare-acquired infections and is a prolific secretor of proteins, including three chitinases (ChiA, ChiB, and ChiC) and a chitin binding protein (Cbp21). In this work, genetic, biochemical, and proteomic approaches identified genes that were required for secretion of all three chitinases and Cbp21. A genetic screen identified a holin-like protein (ChiW) and a putative l-alanyl-d-glutamate endopeptidase (ChiX), and subsequent biochemical analyses established that both were required for nonlytic secretion of the entire chitinolytic machinery, with chitinase secretion being blocked at a late stage in the mutants. In addition, live-cell imaging experiments demonstrated bimodal and coordinated expression of chiX and chiA and revealed that cells expressing chiA remained viable. It is proposed that ChiW and ChiX operate in tandem as components of a protein secretion system used by gram-negative bacteria.


Molecular mechanisms and potential therapeutical targets in Huntington's disease.
Huntington's disease (HD) is a neurodegenerative disorder caused by a CAG repeat expansion in the gene encoding for huntingtin protein. A lot has been learned about this disease since its first description in 1872 and the identification of its causative gene and mutation in 1993. We now know that the disease is characterized by several molecular and cellular abnormalities whose precise timing and relative roles in pathogenesis have yet to be understood. HD is triggered by the mutant protein, and both gain-of-function (of the mutant protein) and loss-of-function (of the normal protein) mechanisms are involved. Here we review the data that describe the emergence of the ancient huntingtin gene and of the polyglutamine trait during the last 800 million years of evolution. We focus on the known functions of wild-type huntingtin that are fundamental for the survival and functioning of the brain neurons that predominantly degenerate in HD. We summarize data indicating how the loss of these beneficial activities reduces the ability of these neurons to survive. We also review the different mechanisms by which the mutation in huntingtin causes toxicity. This may arise both from cell-autonomous processes and dysfunction of neuronal circuitries. We then focus on novel therapeutical targets and pathways and on the attractive option to counteract HD at its primary source, i.e., by blocking the production of the mutant protein. Strategies and technologies used to screen for candidate HD biomarkers and their potential application are presented. Furthermore, we discuss the opportunities offered by intracerebral cell transplantation and the likely need for these multiple routes into therapies to converge at some point as, ideally, one would wish to stop the disease process and, at the same time, possibly replace the damaged neurons.


Skull-fixated fiducial markers improve accuracy in staged frameless stereotactic epilepsy surgery in children.
Surgery to monitor and resect epileptogenic foci may be undertaken in 2 stages, providing an opportunity to use skull-fixated fiducials implanted during the first stage to improve the accuracy of cortical resection during the second stage. This study compared the intrinsic accuracy of skin-based and skull-fixated fiducial markers in registering frameless stereotaxy during pediatric epilepsy surgery. To the authors' knowledge, these modalities of registration have not previously been directly compared in this population. The authors undertook a retrospective review of pediatric patients who underwent resection of epileptogenic foci in 2 stages with frameless stereotactic assistance, performed by a single surgeon at Oregon Health & Science University. For the first stage (subdural grid implantation), 9 skin fiducial markers were used to register anatomical data in a frameless stereotactic station. Intraoperatively, four 3-mm screws were placed circumferentially around the craniotomy. Postoperatively, thin-slice brain MR and CT images were obtained and fused. For the second stage, the 4 screws were used as fiducial markers to register the stereotactic anatomical data. For both stages, accuracy (difference in millimeters from zero of the manual fiducial registration compared with the computer model) was determined using navigation software. The intrinsic accuracy of these 2 methods of fiducial registration was compared using a paired Student t-test. Between 2004 and 2009, 40 pediatric patients with epilepsy underwent frameless stereotactic surgical procedures. Fourteen patients who had 2-stage procedures using skin-based and skull-fixated registration with complete accuracy data were included in this retrospective review. Mean registration error was significantly lower using skull-fixated fiducials (1.35 mm, 95% CI 1.09-1.60 mm) than using skin-based fiducials (1.85 mm, 95% CI 1.56-2.13 mm; p = 0.0016). A significantly higher degree of accuracy was achieved using 4 skull-fixated fiducials compared with using 9 skin-based fiducials. This simple and accurate method for registering frameless stereotactic anatomical data does not involve the potential time, expense, discomfort, and morbidity of extraoperative skull-fixated fiducial placement. The method described in this paper could also be extrapolated to other planned 2-stage cranial surgical procedures such as combined skull base approaches.


The prevalence, course and clinical features of post-concussion syndrome in children.
To investigate the clinical features and the prevalence of symptoms of post-concussion syndrome in children with mild traumatic brain injury, and to evaluate their changes over time. The research involved two groups of 4-16 year-old children: the case group of 301 children who had experienced a single mild traumatic brain injury, and the control group of 301 children who had sustained any other mild body injury without head trauma. Groups were matched according to gender, age, and the date of admission to hospital. In total, 102 matched pairs were analyzed. The period between the date of the trauma and the examination was one to five years (median-27 months). Standardized questionnaires were sent by post to parents from both groups. Parents were asked about the health state as well as symptoms of post-concussion syndrome, their character and frequency. The respondents were inquired about the presence of the symptoms prior to the trauma, shortly after the trauma, and during the last year and the last month before the inquiry. The prevalence of headache, irritability, fears, sleep disorders, learning difficulties, as well as concentration and memory problems did not differ significantly between children with mild traumatic brain injury and the control group when the results of the last year before examination and the last month before the examination were compared. We have investigated how the period of time between the date of the trauma and the date when the questionnaire was filled in influenced the results. The comparison of the questionnaires that were filled a year (but less than two years) after the trauma to those that were filled in 2-5 years after the trauma revealed significant differences in the prevalence of a number of symptoms of the post-concussion syndrome. In children with mild traumatic brain injury, there was a significant decrease in the prevalence of learning difficulties soon after the trauma (p=0.032), headaches before (p=0.026) and soon after the trauma (p=0.01), and irritability the last month before the examination (p=0.043). In children from the control group, there was a significant decrease in the prevalence of concentration problems the last year before examination (p=0.023) and the last month before examination (p=0.036). More than one year after the trauma, the prevalence of the symptoms of the post-concussion syndrome is not significant higher in children with mild traumatic brain injury, compared to children with other mild body injuries, and is comparable by the changes over time.


Topical therapies for rosacea.
Therapeutic options for rosacea include topical agents, oral therapies, laser and light treatments, and surgical procedures. Topical therapies play a critical role in the treatment of patients with papulopustular rosacea and erythematotelangiectatic rosacea, and have the ability to effectively minimize certain manifestations of the disease, including papules, pustules, and erythema. The 3 primary agents for the topical treatment of rosacea are metronidazole, azelaic acid, and sodium sulfacetamide-sulfur. Each of these therapies is approved for the treatment of rosacea and has been validated by multiple studies. Additional topical therapies including benzoyl peroxide, clindamycin, retinoids, topical steroids, calcineurin inhibitors, and permethrin are not approved for the treatment of rosacea and play variable roles in the management of this condition.


Altered regulation of IL-2 production in systemic lupus erythematosus: an evolving paradigm.
In systemic lupus erythematosus (SLE), IL-2 production by T lymphocytes in vitro is impaired. Deficient IL-2 production may be an outcome of a primary SLE T cell disorder that is due to impaired signal transduction. In this issue of the JCI, evidence is presented that an anti-TCR/CD3 complex autoantibody present in SLE sera can bind to T cells and activate the Ca(2+)-calmodulin kinase IV (CaMKIV) signaling cascade, resulting in downregulation of IL-2 transcription and IL-2 production. Because IL-2 may contribute to the maintenance of T cell tolerance, deficient IL-2 production could promote a breach of T cell tolerance that results in autoantibody production in SLE.


HIRA-TAN: a real-time PCR-based system for the rapid identification of causative agents in pneumonia.
Identification of the causative pathogen(s) of pneumonia would allow the selection of effective antibiotics and thus reduce the mortality rate and the emergence of drug-resistant pathogens. To identify such pathogens and to obtain these benefits, it is necessary that a clinical test is rapid, accurate, easily performed, and cost-effective. Here, we devised a PCR-based test, named HIRA-TAN, which is able to discriminate therapeutic targets from commensal organisms (e.g. Streptococcus pneumoniae or Haemophilus influenzae) and to detect foreign organisms (e.g. Mycoplasma pneumoniae or Legionella pneumophila) in the sputum. The utility of this system was validated in a prospective study, using sputum samples from patients with pneumonia. 568 patients were enrolled and the HIRA-TAN assay identified the causative pathogens with an accuracy of 96.7% for H. influenzae; 93.2% for Pseudomonas aeruginosa; 80.6% for Klebsiella pneumoniae; 90.9% for Moraxella catarrhalis; 87.5% for Escherichia coli; 78.1% for MRSA and 91.6% for S. pneumoniae. Overall the HIRA-TAN procedure was able to identify the causative pathogens of pneumonia in 60% of the cases. Additionally, this procedure was able to determine when the pneumonia-causing organism was a commensal organism or a foreign organism in a single assay. The HIRA-TAN approach yielded reproducible results and provided valuable information to plan the course of treatment of pneumonia. Through the rapid identification of the causative pathogens, the HIRA-TAN will promote targeted treatments for pneumonias. UMIN000001694.


Engineered probiotics for local tumor delivery of checkpoint blockade nanobodies.
Checkpoint inhibitors have revolutionized cancer therapy but only work in a subset of patients and can lead to a multitude of toxicities, suggesting the need for more targeted delivery systems. Because of their preferential colonization of tumors, microbes are a natural platform for the local delivery of cancer therapeutics. Here, we engineer a probiotic bacteria system for the controlled production and intratumoral release of nanobodies targeting programmed cell death-ligand 1 (PD-L1) and cytotoxic T lymphocyte-associated protein-4 (CTLA-4) using a stabilized lysing release mechanism. We used computational modeling coupled with experimental validation of lysis circuit dynamics to determine the optimal genetic circuit parameters for maximal therapeutic efficacy. A single injection of this engineered system demonstrated an enhanced therapeutic response compared to analogous clinically relevant antibodies, resulting in tumor regression in syngeneic mouse models. Supporting the potentiation of a systemic immune response, we observed a relative increase in activated T cells, an abscopal effect, and corresponding increases in systemic T cell memory populations in mice treated with probiotically delivered checkpoint inhibitors. Last, we leveraged the modularity of our platform to achieve enhanced therapeutic efficacy in a poorly immunogenic syngeneic mouse model through effective combinations with a probiotically produced cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF). Together, these results demonstrate that our engineered probiotic system bridges synthetic biology and immunology to improve upon checkpoint blockade delivery.


Low-energy electron holography imaging of conformational variability of single-antibody molecules from electrospray ion beam deposition.
Imaging of proteins at the single-molecule level can reveal conformational variability, which is essential for the understanding of biomolecules. To this end, a biologically relevant state of the sample must be retained during both sample preparation and imaging. Native electrospray ionization (ESI) can transfer even the largest protein complexes into the gas phase while preserving their stoichiometry and overall shape. High-resolution imaging of protein structures following native ESI is thus of fundamental interest for establishing the relation between gas phase and solution structure. Taking advantage of low-energy electron holography's (LEEH) unique capability of imaging individual proteins with subnanometer resolution, we investigate the conformational flexibility of Herceptin, a monoclonal IgG antibody, deposited by native electrospray mass-selected ion beam deposition (ES-IBD) on graphene. Images reconstructed from holograms reveal a large variety of conformers. Some of these conformations can be mapped to the crystallographic structure of IgG, while others suggest that a compact, gas-phase-related conformation, adopted by the molecules during ES-IBD, is retained. We can steer the ratio of those two types of conformations by changing the landing energy of the protein on the single-layer graphene surface. Overall, we show that LEEH can elucidate the conformational heterogeneity of inherently flexible proteins, exemplified here by IgG antibodies, and thereby distinguish gas-phase collapse from rearrangement on surfaces.


Complete genome sequence of bile-isolated Enterococcus avium strain 352.
Enterococcus avium is a Gram-positive pathogenic bacterium belonging to the family Enterobacteriaceae. E. avium can cause bacteremia, peritonitis, and intracranial suppurative infection. However, the mechanism of its pathogenesis and its adaptation to a special niche is still unclear. In this study, the E. avium strain 352 was isolated from human bile and whole genome sequencing was performed. The E. avium strain 352 consists of a circular 4,794,392 bp chromosome as well as an 87,705 bp plasmid. The GC content of the chromosome is 38.98%. There are 4905 and 99 protein coding sequences in the chromosome and the plasmid, respectively. The genome of the E. avium strain 352 contains number of genes reported to be associated with bile adaption, including bsh, sbcC, mutS, nifI, galU, and hupB. There are also several virulence-associated genes including esp, fss1, fss3, ecbA, bsh, lap, clpC, clpE, and clpP. This study demonstrates the presence of various virulence factors of the E. avium strain 352, which has the potential to cause infections. Moreover, the genes involved in bile adaption might contribute to its ability to live in bile. Further comparative genomic studies would help to elucidate the evolution of pathogenesis of E. avium.


Prevalence of alcohol dependence among US adult drinkers, 2009-2011.
Excessive alcohol consumption is responsible for 88,000 deaths annually and cost the United States $223.5 billion in 2006. It is often assumed that most excessive drinkers are alcohol dependent. However, few studies have examined the prevalence of alcohol dependence among excessive drinkers. The objective of this study was to update prior estimates of the prevalence of alcohol dependence among US adult drinkers. Data were analyzed from the 138,100 adults who responded to the National Survey on Drug Use and Health in 2009, 2010, or 2011. Drinking patterns (ie, past-year drinking, excessive drinking, and binge drinking) were assessed by sociodemographic characteristics and alcohol dependence (assessed through self-reported survey responses and defined as meeting ≥3 of 7 criteria for dependence in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition). Excessive drinking, binge drinking, and alcohol dependence were most common among men and those aged 18 to 24. Binge drinking was most common among those with annual family incomes of $75,000 or more, whereas alcohol dependence was most common among those with annual family incomes of less than $25,000. The prevalence of alcohol dependence was 10.2% among excessive drinkers, 10.5% among binge drinkers, and 1.3% among non-binge drinkers. A positive relationship was found between alcohol dependence and binge drinking frequency. Most excessive drinkers (90%) did not meet the criteria for alcohol dependence. A comprehensive approach to reducing excessive drinking that emphasizes evidence-based policy strategies and clinical preventive services could have an impact on reducing excessive drinking in addition to focusing on the implementation of addiction treatment services.


Penicillinase producing strains of Neisseria gonorrhoeae in the West Midlands 1979-83.
All cases of gonorrhoea caused by penicillinase producing strains of Neisseria gonorrhoeae (PPNG) occurring in the population served by the West Midlands Regional Health Authority were surveyed. Isolates that were subjected to plasmid analysis were found to be mainly the "Asian" type in travellers from abroad and their immediate contacts, whereas isolates apparently indigenous in origin were mainly "African" type. The emerging sigmoid curve of yearly incidence, which is apparent for both the country and the region, may indicate that a dynamic equilibrium is being approached in the susceptible core populations. The importance of vigorous contact tracing is underlined by the contrast between the incidence of PPNG strains in the United Kingdom and the larger numbers found in areas where they are hyperendemic and where contact tracing is ineffectual or non-existent.


Discrete ephrin-B1 expression by specific layers of the primate retinogeniculostriate system continues throughout postnatal and adult life.
The molecular guidance cue ephrin-B1 has traditionally been associated with the early development of the visual system, encompassing retinocollicular mapping as well as development and maturation of synapses. Although little is known about its role in the visual system during the postnatal period and in adulthood, recent studies have demonstrated the expression of ephrin-B1 in the adult mouse brain, indicating a sustained role beyond early development. Therefore, we explored the spatiotemporal expression of ephrin-B1 in the postnatal and adult nonhuman primate visual system and demonstrated that a modulated expression continued following birth into adulthood in the lateral geniculate nucleus (LGN) and primary visual cortex (V1, striate cortex). This occurred in the layers involved in bidirectional geniculostriate communication: layers 3Bβ, 4, and 6 of V1 and the parvocellular (P) and magnocellular (M) layers of the LGN. Furthermore, discrete gradients between the ipsi- and contralateral inputs of the P and M layers of the LGN evolved between 1 month following birth and the start of the critical period (3 months), and continued into adulthood. We also detected the postsynaptic expression of ephrin-B1 by excitatory cells in adult LGN and V1 and a subset of interneurons in adult V1, suggestive of a more global rather than subtype-specific role. Together these results suggest a possible role for ephrin-B1 in the maturation of the primate retinogeniculostriate pathway throughout postnatal life, extending into adulthood.


Mycobacterium avium complex pulmonary disease: new epidemiology and management concepts.
The prevalence of Mycobacterium avium complex (MAC)-related pulmonary disease has been increasing because of environmental factors, changes in organism virulence, and evolving host susceptibility. Treatment is often complicated by adverse effects, development of drug resistance, and refractory disease, with recurrence rates as high as 25-45%. Aerosolization of water, soil, or dusts are the likely sources of MAC-related pulmonary disease in susceptible individuals. The management of MAC-related pulmonary disease requires a multimodality approach, including antimicrobial therapy in appropriate patients, employment of mucus clearance techniques, instituting changes in the individual's home environment and personal habits to reduce environmental exposure to MAC, prevention of reflux, and maintenance of a healthy body weight. When the standard treatment for MAC-related pulmonary disease is not possible because of drug intolerance, antibiotic resistance, or progression of disease, second-line agents such as inhaled amikacin, clofazimine, bedaquiline, and delamanid must be considered, despite limited experience and few studies to guide their use. Individuals who have proven to be susceptible to MAC-related pulmonary disease should institute measures to reduce exposure to environmental sources of infection. Further research is needed to assess the impact of such preventive strategies on the incidence of new infection and disease recurrence. The efficacy of new medications for MAC-related pulmonary disease and their use in different combinations also requires further study.


Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling.
Diffuse large B-cell lymphoma (DLBCL), the most common subtype of non-Hodgkin's lymphoma, is clinically heterogeneous: 40% of patients respond well to current therapy and have prolonged survival, whereas the remainder succumb to the disease. We proposed that this variability in natural history reflects unrecognized molecular heterogeneity in the tumours. Using DNA microarrays, we have conducted a systematic characterization of gene expression in B-cell malignancies. Here we show that there is diversity in gene expression among the tumours of DLBCL patients, apparently reflecting the variation in tumour proliferation rate, host response and differentiation state of the tumour. We identified two molecularly distinct forms of DLBCL which had gene expression patterns indicative of different stages of B-cell differentiation. One type expressed genes characteristic of germinal centre B cells ('germinal centre B-like DLBCL'); the second type expressed genes normally induced during in vitro activation of peripheral blood B cells ('activated B-like DLBCL'). Patients with germinal centre B-like DLBCL had a significantly better overall survival than those with activated B-like DLBCL. The molecular classification of tumours on the basis of gene expression can thus identify previously undetected and clinically significant subtypes of cancer.


Regularized Speaker Adaptation of KL-HMM for Dysarthric Speech Recognition.
This paper addresses the problem of recognizing the speech uttered by patients with dysarthria, which is a motor speech disorder impeding the physical production of speech. Patients with dysarthria have articulatory limitation, and therefore, they often have trouble in pronouncing certain sounds, resulting in undesirable phonetic variation. Modern automatic speech recognition systems designed for regular speakers are ineffective for dysarthric sufferers due to the phonetic variation. To capture the phonetic variation, Kullback-Leibler divergence-based hidden Markov model (KL-HMM) is adopted, where the emission probability of state is parameterized by a categorical distribution using phoneme posterior probabilities obtained from a deep neural network-based acoustic model. To further reflect speaker-specific phonetic variation patterns, a speaker adaptation method based on a combination of L2 regularization and confusion-reducing regularization, which can enhance discriminability between categorical distributions of the KL-HMM states while preserving speaker-specific information is proposed. Evaluation of the proposed speaker adaptation method on a database of several hundred words for 30 speakers consisting of 12 mildly dysarthric, 8 moderately dysarthric, and 10 non-dysarthric control speakers showed that the proposed approach significantly outperformed the conventional deep neural network-based speaker adapted system on dysarthric as well as non-dysarthric speech.


Structural and electronic properties of bulk and low-index surfaces of zincblende PtC.
Transition metal carbides have been extensively used in diverse applications over the past decade. Their versatility is in part thanks to their unique bonding, which displays a mixture of ionic, metallic and covalent character. While the bulk structure of zincblende (ZB) PtC has been investigated several times, a detailed understanding of the electronic and structural properties of its low-index surfaces is lacking. In this work, we present an ab initio investigation of the properties of five crystallographic ZB PtC surfaces (Pt/C-terminated PtC(1 0 0), PtC(1 1 0) and Pt/C-terminated PtC(1 1 1)). Upon geometry optimization, both polar and nonpolar surfaces undergo a mild interlayer relaxation, without extensive reconstructions. Calculated vacancy formation energies indicate facile C removal on the (1 1 1) surface while Pt-vacancy formation is endothermic. Finally, atomic O adsorption energies on all surfaces reveal a high affinity of the C-terminated surfaces towards this species.


Induction of a melanoma-specific antibody response by a monovalent, but not a divalent, synthetic GM2 neoglycopeptide.
The GM2 ganglioside represents an important target for specific anticancer immunotherapy. We designed and synthesized a neoglycopeptide immunogen displaying one or two copies of the GM2 tetrasaccharidic moiety. These glycopeptides were prepared using the Huisgen cycloaddition, which enables the efficient ligation of the alkyne-functionalized biosynthesized GM2 with an azido CD4(+) T cell epitope peptide. It is worth noting that the GM2 can be produced on a gram scale in bacteria, which can be advantageous for a scale-up of the process. We show here for the first time that a fully synthetic glycopeptide, which is based on a ganglioside carbohydrate moiety, can induce human tumor cell-specific antibodies after immunization in mice. Interestingly, the monovalent, but not the divalent, form of GM2 peptide construct induced antimelanoma antibodies. Unlike traditional vaccines, this vaccine is a pure chemically-defined entity, a key quality for consistent studies and safe clinical evaluation. Therefore, such carbohydrate-peptide conjugate represents a promising cancer vaccine strategy for active immunotherapy targeting gangliosides.


Genitourinary syndrome of menopause: effects on related factors, quality of life, and self-care power.
This study aimed to determine the effects of genitourinary syndrome of menopause on related factors, quality of life, and self-care power. This case-control epidemiological study included 300 postmenopausal women aged 48-60 years who were divided into two groups: the case group (141) with genitourinary syndrome and the control group (159) without genitourinary syndrome. Data were evaluated by chi-square and t-tests in independent groups and logistic regression analysis. The incidence of genitourinary syndrome of menopause was 0.61 times higher in women with a lower educational level than in those with a high educational level, 0.44 times higher in those with chronic disease than in those without, 2.54 times higher in those who continuously used medication than in those who did not, and 2.45 times higher in those with urinary incontinence than in those without. The mean Self-Care Power Scale score was significantly higher in the control group (101.52 ±21.50) than in the case group (83.96 ±25.64). The mean Menopause-Specific Quality of Life Scale score was significantly higher in the case group (65.52 ±33.47) than in the control group (37.06 ±25.81). Low educational level, chronic disease, urinary incontinence, and continuous drug use are associated with genitourinary syndrome of menopause, which negatively affects the quality of life and self-care power in women.


Cognition assessment using the NIH Toolbox.
Cognition is 1 of 4 domains measured by the NIH Toolbox for the Assessment of Neurological and Behavioral Function (NIH-TB), and complements modules testing motor function, sensation, and emotion. On the basis of expert panels, the cognition subdomains identified as most important for health, success in school and work, and independence in daily functioning were Executive Function, Episodic Memory, Language, Processing Speed, Working Memory, and Attention. Seven measures were designed to tap constructs within these subdomains. The instruments were validated in English, in a sample of 476 participants ranging in age from 3 to 85 years, with representation from both sexes, 3 racial/ethnic categories, and 3 levels of education. This report describes the development of the Cognition Battery and presents results on test-retest reliability, age effects on performance, and convergent and discriminant construct validity. The NIH-TB Cognition Battery is intended to serve as a brief, convenient set of measures to supplement other outcome measures in epidemiologic and longitudinal research and clinical trials. With a computerized format and national standardization, this battery will provide a "common currency" among researchers for comparisons across a wide range of studies and populations.


HSP70/HSF1 axis, regulated via a PI3K/AKT pathway, is a druggable target in chronic lymphocytic leukemia.
Considering the role played by the heat shock protein of 70 kDa (HSP70) in cancer, we characterized this protein and its major regulator, the heat shock factor 1 (HSF1), in chronic lymphocytic leukemia (CLL). We found both HSP70 and HSF1 overexpressed in CLL patients, correlated to poor prognosis and abnormally localized in the nucleus of leukemic B cells. The two proteins were strictly correlated each other and their levels decreased consensually in those patients responding to in vivo therapeutic regimens. HSP70 and HSF1 inhibition was proved to be effective in inducing a dose-dependent in vitro apoptosis of CLL B cells. Considering that HSF1 is finely regulated by kinases belonging to pathways triggered by rat sarcoma (RAS), we benefited from a previous proteomic study performed in CLL patients aiming to assess the activation/expression of key signaling proteins. We found that patients showing high levels of HSP70 also expressed high Akt-Ser473, thus activating HSF1. Inhibition of PI3K, which activates AKT, reduced the expression of HSF1 and HSP70. By contrast, HSP70-low patients displayed high activation of MEK1/2 and ERK1/2, known to negatively regulate HSF1. These data demonstrate that the HSP70 expression is regulated by the modulation of HSF1 activity through the activation of RAS-regulated pathways and suggest the HSP70/HSF1 interplay as an interesting target for antileukemic therapies. Finally, inhibition of PI3K, that activates AKT, reduced the expression of HSF1 and HSP70.


The homeobox transcription factor Even-skipped regulates acquisition of electrical properties in Drosophila neurons.
While developmental processes such as axon pathfinding and synapse formation have been characterized in detail, comparatively less is known of the intrinsic developmental mechanisms that regulate transcription of ion channel genes in embryonic neurons. Early decisions, including motoneuron axon targeting, are orchestrated by a cohort of transcription factors that act together in a combinatorial manner. These transcription factors include Even-skipped (Eve), islet and Lim3. The perdurance of these factors in late embryonic neurons is, however, indicative that they might also regulate additional aspects of neuron development, including the acquisition of electrical properties. To test the hypothesis that a combinatorial code transcription factor is also able to influence the acquisition of electrical properties in embryonic neurons we utilized the molecular genetics of Drosophila to manipulate the expression of Eve in identified motoneurons. We show that increasing expression of this transcription factor, in two Eve-positive motoneurons (aCC and RP2), is indeed sufficient to affect the electrical properties of these neurons in early first instar larvae. Specifically, we observed a decrease in both the fast K+ conductance (IKfast) and amplitude of quantal cholinergic synaptic input. We used charybdotoxin to pharmacologically separate the individual components of IKfast to show that increased Eve specifically down regulates the Slowpoke (a BK Ca2+-gated potassium channel), but not Shal, component of this current. Identification of target genes for Eve, using DNA adenine methyltransferase identification, revealed strong binding sites in slowpoke and nAcRalpha-96Aa (a nicotinic acetylcholine receptor subunit). Verification using real-time PCR shows that pan-neuronal expression of eve is sufficient to repress transcripts for both slo and nAcRalpha-96Aa. Taken together, our findings demonstrate, for the first time, that Eve is sufficient to regulate both voltage- and ligand-gated currents in motoneurons, extending its known repertoire of action beyond its already characterized role in axon guidance. Our data are also consistent with a common developmental program that utilizes a defined set of transcription factors to determine both morphological and functional neuronal properties.


Monitoring and evaluating the impact of national school-based deworming in Kenya: study design and baseline results.
An increasing number of countries in Africa and elsewhere are developing national plans for the control of neglected tropical diseases. A key component of such plans is school-based deworming (SBD) for the control of soil-transmitted helminths (STHs) and schistosomiasis. Monitoring and evaluation (M&E) of national programmes is essential to ensure they are achieving their stated aims and to evaluate when to reduce the frequency of treatment or when to halt it altogether. The article describes the M&E design of the Kenya national SBD programme and presents results from the baseline survey conducted in early 2012. The M&E design involves a stratified series of pre- and post-intervention, repeat cross-sectional surveys in a representative sample of 200 schools (over 20,000 children) across Kenya. Schools were sampled based on previous knowledge of STH endemicity and were proportional to population size. Stool (and where relevant urine) samples were obtained for microscopic examination and in a subset of schools; finger-prick blood samples were collected to estimate haemoglobin concentration. Descriptive and spatial analyses were conducted. The evaluation measured both prevalence and intensity of infection. Overall, 32.4% of children were infected with at least one STH species, with Ascaris lumbricoides as the most common species detected. The overall prevalence of Schistosoma mansoni was 2.1%, while in the Coast Province the prevalence of S. haematobium was 14.8%. There was marked geographical variation in the prevalence of species infection at school, district and province levels. The prevalence of hookworm infection was highest in Western Province (25.1%), while A. lumbricoides and T. trichiura prevalence was highest in the Rift Valley (27.1% and 11.9%). The lowest prevalence was observed in the Rift Valley for hookworm (3.5%), in the Coast for A. lumbricoides (1.0%), and in Nyanza for T. trichiura (3.6%). The prevalence of S. mansoni was most common in Western Province (4.1%). The current findings are consistent with the known spatial ecology of STH and schistosome infections and provide an important empirical basis on which to evaluate the impact of regular mass treatment through the school system in Kenya.


A cross-sectional study on obesity and related risk factors among women of the central market of Lusonga in Lubumbashi, Democratic Republic of Congo.
Obesity is known as one of adjuvant factors for increase in non-communiable diseases (NCDs). The aim of this study was to describe the prevalence of obesity and identify its risk factors among women of the central market of Lusonga in Lubumbashi, Democratic Republic of Congo. In October 2014, we interviewed a total of 430 women selling in the central market of Lusonga in Lubumbashi. Data on sociodemographic characteristics, health-related habits and behaviors, diet, physical activity, chronic diseases, blood pressure and anthropometric measurements were collected. A multivariate logistic regression model was fitted. Prevalence of overweight and obesity was 16.51% and 13.26% respectively. The logistic regression did not show any significant association between age and obesity. Risk of obesity was lower in married women (adjusted odds ratio (aOR) = 0.23 (0.08-0.63)). Women with low educational profile (primary school or less) were more likely to be obese than those with higher education (secondary or high school) (aOR = 2.50 (1.12-5.63)). Risk of obesity increased with living in urban area (aOR = 2.52 (1.00-6.36)), use of oral birth control pills (aOR = 11.07 (3.52-34.83)) and low consumption of fruit (aOR = 5.47 (1.88-15.92)) and vegetable (aOR = 2.42 (1.05-5.56)). Obese women were more likely to be hypertensive than non-obese (aOR = 7.15 (2.46-20.75)) and diabetics (aOR = 3.62 (1.62-8.11)). This study has reported a prevalence of 13.26% of obesity among women selling at Lusonga's market. Marital status, education level, residence, use of oral birth control pills and consumption of fruit and vegetables had a significant association with the prevalence of obesity in this category of women.


Toward precision medicine for preserving fertility in cancer patients: existing and emerging fertility preservation options for women.
As the number of young cancer survivors increases, quality of life after cancer treatment is becoming an ever more important consideration. According to a report from the American Cancer Society, approximately 810,170 women were diagnosed with cancer in 2015 in the United States. Among female cancer survivors, 1 in 250 are of reproductive age. Anticancer therapies can result in infertility or sterility and can have long-term negative effects on bone health, cardiovascular health as a result of reproductive endocrine function. Fertility preservation has been identified by many young patients diagnosed with cancer as second only to survival in terms of importance. The development of fertility preservation technologies aims to help patients diagnosed with cancer to preserve or protect their fertility prior to exposure to chemo- or radiation therapy, thus improving their chances of having a family and enhancing their quality of life as a cancer survivor. Currently, sperm, egg, and embryo banking are standard of care for preserving fertility for reproductive-age cancer patients; ovarian tissue cryopreservation is still considered experimental. Adoption and surrogate may also need to be considered. All patients should receive information about the fertility risks associated with their cancer treatment and the fertility preservation options available in a timely manner, whether or not they decide to ultimately pursue fertility preservation. Because of the ever expanding number of options for treating cancer and preserving fertility, there is now an opportunity to take a precision medicine approach to informing patients about the fertility risks associated with their cancer treatment and the fertility preservation options that are available to them.


Tris(2-pyridyl)phosphine as a versatile ligand for pnictogen acceptors.
We report cationic complexes of arsenic and antimony with the tris(2-pyridyl)phosphine ligand. Chloride ion abstraction from AsCl3 using TMSOTf in the presence of the ligand gives [P(Pyr)3As][OTf]3, in which the trication adopts a C3v symmetric cage structure. The reaction proceeds via the intermediate [P(Pyr)3AsCl][OTf]2, which undergoes chloride exchange to give [P(Pyr)3As][OTf]3 and [P(Pyr)3AsCl2][OTf]. The rearrangement reaction has been supported by the isolation of the antimony mono fluoride derivative [P(Pyr)3SbF][OTf]2. The asymmetric axial lone pairs in derivatives of [P(Pyr)3Pn]3+ are electronically separated. The HOMO-1 (for arsenic) and HOMO (for antimony) represent the major contribution to the phosphine lone pair indicating the possibility for nucleophilic behaviour despite the +3 charge. Less accessible is the HOMO-7, which represents the lone pair at arsenic or antimony, respectively.


Role of neuroimaging in analgesic drug development.
Rapidly developing, non-invasive, neuroimaging methods provide increasingly detailed structural and functional information about the nervous system, helping advance our understanding of pain processing, chronic pain conditions and the mechanisms of analgesia. However, effective treatment for many chronic pain conditions remains a large, unmet medical need. Neuroimaging techniques may enhance our understanding of why currently available analgesics are ineffective for so many patients and aid in identifying new neural targets for pharmacological interventions of pain. This review examines how neuroimaging has enhanced our understanding of the mechanisms of chronic pain, the neural correlates of pharmacological modulation of pain, and the role of neuroimaging in analgesic development. Rather than focusing on one method, we discuss the advantages and limitations of several techniques that may each serve a unique role in aiding drug development, and we discuss current issues that exist in the design and implementation of pharmacological neuroimaging studies. Particularly, experimental design must be carefully considered as there are limitations in terms of the pharmacokinetics of the drug of interest as well as in respect to the capabilities of the neuroimaging method in use. Finally, we identify future directions including novel approaches that may also play a role in furthering our knowledge of the neural basis of analgesia. In the future, neuroimaging will certainly impact the methodology of analgesic drug development as it may lead to quicker and more efficient methods of evaluating the neural modulation of chronic pain.


Functional analyses reveal extensive RRE plasticity in primary HIV-1 sequences selected under selective pressure.
HIV-1 Rev response element (RRE) is a functional region of viral RNA lying immediately downstream to the junction of gp120 and gp41 in the env coding sequence. The RRE is essential for HIV replication and binds with the Rev protein to facilitate the export of viral mRNA from nucleus to cytoplasm. It has been suggested that changes in the predicted secondary structure of primary RRE sequences impact the function of the RREs; however, functional assays have not yet been performed. The aim of this study was to characterize the genetic, structural and functional variation in the RRE primary sequences selected in vivo by Enfuvirtide pressure. Multiple RRE variants were obtained from viruses isolated from patients who failed an Enfuvirtide-containing regimen. Different alterations were observed in the predicted RRE secondary structures, with the abrogation of the primary Rev binding site in one of the variants. In spite of this, most of the RRE variants were able to bind Rev and promote the cytoplasmic export of the viral mRNAs with equivalent efficiency in a cell-based assay. Only RRE45 and RRE40-45 showed an impaired ability to bind Rev in a gel-shift binding assay. Unexpectedly, this impairment was not reflected in functional capacity when RNA export was evaluated using a reporter assay, or during virus replication in lymphoid cells, suggesting that in vivo the RRE would be highly malleable. The Rev-RRE functionality is unaffected in RRE variants selected in patients failing an ENF-containing regimen. Our data show that the current understanding of the Rev-RRE complex structure does not suffice and fails to rationally predict the function of naturally occurring RRE mutants. Therefore, this data should be taken into account in the development of antiviral agents that target the RRE-Rev complex.


A Prospective Study of Community Mediators on the Risk of Sepsis After Cancer.
Few studies have examined whether community factors mediate the relationship between patients surviving cancer and future development of sepsis. We determined the influence of community characteristics upon risk of sepsis after cancer, and whether there are differences by race. We performed a prospective analysis using data from the REasons for Geographic and Racial Differences in Stroke cohort years 2003 to 2012 complemented with county-level community characteristics from the American Community Survey and County Health Rankings. We categorized those with a self-reported prior cancer diagnosis as "cancer survivors" and those without a history of cancer as "no cancer history." We defined sepsis as hospitalization for a serious infection with ≥2 systemic inflammatory response syndrome criteria. We examined the mediation effect of community characteristics on the association between cancer survivorship and sepsis incidence using Cox proportional hazards models adjusted for age, sex, race, and total number of comorbidities. We repeated analysis stratified by race. There were 28 840 eligible participants, of which 2860 (9.92%) were cancer survivors, and 25 289 (90.08%) were no cancer history participants. The only observed community-level mediation effects were from income (% mediated 0.07%; natural indirect effect [NIE] on hazard scale] = 1.001, 95% confidence interval [95% CI]: 1.000-1.005) and prevalence of adult smoking (% mediated = 0.21%; NIE = 1.002, 95% CI: 1.000-1.004). We observed similar effects when stratified by race. Cancer survivors are at increased risk of sepsis; however, this association is weakly mediated by community poverty and smoking prevalence.


Effect of silage botanical composition on ruminal biohydrogenation and transfer of fatty acids to milk in dairy cows.
Ruminal biohydrogenation and transfer of fatty acids (FA) to milk were determined for 4 silages with different botanical compositions using 4 multiparous Norwegian Red dairy cows [(mean ± SD) 118 ± 40.9 d in milk, 22.5 ± 2.72 kg of milk/d, 631 ± 3.3 kg of body weight, 3.3 ± 0.40 points on body condition score at the start of the experiment] fitted with rumen cannulas. Treatments consisted of 4 experimental silages: a mix of the first and third cut of organically managed short-term grassland with timothy (Phleum pratense L.) and red clover (Trifolium pratense L.; 2 yr old; ORG-SG); organically managed long-term grassland with a high proportion of unsown species (6 yr old; ORG-LG); conventionally managed ley with perennial ryegrass (Lolium perenne L.; CON-PR); and conventionally managed ley with timothy (CON-TI). The herbages were cut, wilted, and preserved with additive in round bales and fed at 0.90 of ad libitum intake. A barley (Hordeum vulgare L.) concentrate constituted 300 g/kg of dry matter of the total feed offered. A Latin square design (4 × 4) with 3-wk periods and the last week in each period used for sampling was implemented. Omasal flows of FA were measured using Yb acetate, Cr-EDTA, and the indigestible neutral detergent fiber fraction as indigestible markers. The composition of FA was analyzed in feed, omasal digesta, and milk. Compared with ORG-LG, ORG-SG had a higher herbage proportion of red clover (0.36 vs. 0.01) and lower proportions of timothy (0.42 vs. 0.18), smooth meadowgrass (Poa pratensis L.), meadow fescue (Festuca pratensis Huds.), white clover (Trifolium repens L.), dandelion (Taraxacum spp.), and creeping buttercup (Ranunculus repens L.). The silages were well preserved. The concentration of neutral detergent fiber was higher and the concentration of Kjeldahl-N was lower for CON-TI than for the other silages. Silage type had no effect on dry matter intake, but milk yield was lower for CON-TI than for the other silages. Apparent biohydrogenation of C18:3n-3 was lower for ORG-SG (932 g/kg) than for ORG-LG (956 g/kg), CON-PR (959 g/kg), and CON-TI (958 g/kg). Compared with the grass-based silages, ORG-SG and ORG-LG resulted in higher omasal flows of C18:1 trans FA and higher milk fat proportions of C18:1 trans FA and C18:2 cis-9,trans-11. Apparent recovery of C18:3n-3 in milk was higher for ORG-SG (61 g/kg) than for ORG-LG (33 g/kg), CON-PR (34 g/kg), and CON-TI (38 g/kg), and milk fat proportion of C18:3n-3 was higher for ORG-SG than for CON-TI. Milk fat proportions of C16:0 were lower for ORG-SG and ORG-LG compared with those for CON-PR and CON-TI. It was concluded that high proportions of red clover and other dicotyledons in the silages affected ruminal biohydrogenation and increased milk fat proportions of beneficial FA.


Subunit interactions within the Saccharomyces cerevisiae DNA polymerase epsilon (pol epsilon ) complex. Demonstration of a dimeric pol epsilon.
Saccharomyces cerevisiae DNA polymerase epsilon (pol epsilon) is essential for chromosomal replication. A major form of pol epsilon purified from yeast consists of at least four subunits: Pol2p, Dpb2p, Dpb3p, and Dpb4p. We have investigated the protein/protein interactions between these polypeptides by using expression of individual subunits in baculovirus-infected Sf9 insect cells and by using the yeast two-hybrid assay. The essential subunits, Pol2p and Dpb2p, interact directly in the absence of the other two subunits, and the C-terminal half of POL2, the only essential portion of Pol2p, is sufficient for interaction with Dpb2p. Dpb3p and Dpb4p, non-essential subunits, also interact directly with each other in the absence of the other two subunits. We propose that Pol2p.Dpb2p and Dpb3p.Dpb4p complexes interact with each other and document several interactions between individual members of the two respective complexes. We present biochemical evidence to support the proposal that pol epsilon may be dimeric in vivo. Gel filtration of the Pol2p.Dpb2p complexes reveals a novel heterotetrameric form, consisting of two heterodimers of Pol2p.Dpb2p. Dpb2p, but not Pol2p, exists as a homodimer, and thus the Pol2p dimerization may be mediated by Dpb2p. The pol2-E and pol2-F mutations that cause replication defects in vivo weaken the interaction between Pol2p and Dpb2p and also reduce dimerization of Pol2p. This suggests, but does not prove, that dimerization may also occur in vivo and be essential for DNA replication.


Treatment of Lyme disease.
We compared phenoxymethyl penicillin, erythromycin, and tetracycline, in each instance 250 mg four times a day for 10 days, for the treatment of early Lyme disease (stage 1). None of 39 patients given tetracycline developed major late complications compared with 3 of 40 penicillin-treated patients and 4 of 29 given erythromycin (p = 0.07). However, with all three antibiotic agents, nearly half of patients had minor late symptoms. For neurologic abnormalities (stage 2), 12 patients were treated with high-dose intravenous penicillin, 20 million U a day for 10 days. Pain usually subsided during therapy, but a mean of 7 to 8 weeks was required for complete recovery of motor deficits. For the treatment of established arthritis (stage 3), 20 patients were assigned treatment with intramuscular benzathine penicillin (7.2 million U) and 20 patients received saline. Seven of the 20 penicillin-treated patients (35%) were apparently cured, but all 20 patients given placebo continued to have attacks of arthritis (P less than 0.02). Of 20 arthritis patients treated with intravenous penicillin G, 20 million U a day for 10 days, 11 (55%) were apparently cured. Thus, all 3 stages of Lyme disease can be treated with antibiotic therapy, but some patients with late disease may not respond.


Estimation of coronary risk factors in British schoolchildren: a preliminary report.
Surveys from several countries have identified the presence of risk factors known to be associated with coronary heart disease in children. Data on the distribution of coronary risk factor variables in British children are scarce. This study was therefore designed to test the feasibility of collecting coronary risk factor data from British children and to conduct a preliminary examination of the problem. One hundred and seven children (mean age 12.8 yr) had their height, weight, triceps skinfold and blood pressure checked. Blood samples for cholesterol and HDL-cholesterol analysis were obtained from 93 children. Peak VO2 was determined on 48 children, 76 children had their daily activity monitored and 59 children's stage of sexual maturity was assessed. The boys' peak VO2 was significantly higher than the girls, whether expressed in l.min-1 (p less than 0.05) or ml.kg.-1min-1 (p less than 0.01). No other significant differences (p greater than 0.05) between the sexes were detected. The results indicate that children have relatively high serum cholesterol levels (boys 4.58 +/- 0.79; girls 4.72 +/- 0.80 mmol.l-1). The willingness and enthusiasm of the children, parents and schools to take part in the study clearly demonstrated the feasibility of a large scale study being successfully pursued in the United Kingdom.


Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data.
Whole-genome bisulfite sequencing (WGBS) is becoming an increasingly accessible technique, used widely for both fundamental and disease-oriented research. Library preparation methods benefit from a variety of available kits, polymerases and bisulfite conversion protocols. Although some steps in the procedure, such as PCR amplification, are known to introduce biases, a systematic evaluation of biases in WGBS strategies is missing. We perform a comparative analysis of several commonly used pre- and post-bisulfite WGBS library preparation protocols for their performance and quality of sequencing outputs. Our results show that bisulfite conversion per se is the main trigger of pronounced sequencing biases, and PCR amplification builds on these underlying artefacts. The majority of standard library preparation methods yield a significantly biased sequence output and overestimate global methylation. Importantly, both absolute and relative methylation levels at specific genomic regions vary substantially between methods, with clear implications for DNA methylation studies. We show that amplification-free library preparation is the least biased approach for WGBS. In protocols with amplification, the choice of bisulfite conversion protocol or polymerase can significantly minimize artefacts. To aid with the quality assessment of existing WGBS datasets, we have integrated a bias diagnostic tool in the Bismark package and offer several approaches for consideration during the preparation and analysis of WGBS datasets.


Prolonged intubation injuries of the larynx: endoscopic diagnosis, classification, and treatment.
Laryngeal trauma from prolonged endotracheal intubation occurs in patients of all ages. Most changes are superficial and heal quickly. Injuries that are found consistently during intubation include nonspecific changes, edema, granulation tissue, ulceration, and other miscellaneous injuries. In this paper significant, severe, and lasting trauma of the larynx has been classified on the basis of the known factors in pathogenesis, observations made at endoscopy, and photographic documentation. This classification has required introduction of new descriptive terminology: "tongues of granulation tissue," "ulcerated troughs," "healed furrows," and "healed fibrous nodule." During intubation the degree of injury can be precisely assessed under general anesthesia by using telescopes for image magnification, thus assisting a decision whether to continue intubation or perform tracheotomy to minimize long-term morbidity. Changes that are found after extubation result from granulation tissue, ulceration, or a combination of both and have been illustrated on flow charts; a knowledge and understanding of these sequelae allows them to be identified by both indirect and direct laryngoscopy so that treatment can be planned.


Plasmodium falciparum PfSET7: enzymatic characterization and cellular localization of a novel protein methyltransferase in sporozoite, liver and erythrocytic stage parasites.
Epigenetic control via reversible histone methylation regulates transcriptional activation throughout the malaria parasite genome, controls the repression of multi-copy virulence gene families and determines sexual stage commitment. Plasmodium falciparum encodes ten predicted SET domain-containing protein methyltransferases, six of which have been shown to be refractory to knock-out in blood stage parasites. We have expressed and purified the first recombinant malaria methyltransferase in sufficient quantities to perform a full enzymatic characterization and reveal the ill-defined PfSET7 is an AdoMet-dependent histone H3 lysine methyltransferase with highest activity towards lysines 4 and 9. Steady-state kinetics of the PfSET7 enzyme are similar to previously characterized histone methyltransferase enzymes from other organisms, however, PfSET7 displays specific protein substrate preference towards nucleosomes with pre-existing histone H3 lysine 14 acetylation. Interestingly, PfSET7 localizes to distinct cytoplasmic foci adjacent to the nucleus in erythrocytic and liver stage parasites, and throughout the cytoplasm in salivary gland sporozoites. Characterized recombinant PfSET7 now allows for target based inhibitor discovery. Specific PfSET7 inhibitors can aid in further investigating the biological role of this specific methyltransferase in transmission, hepatic and blood stage parasites, and may ultimately lead to the development of suitable antimalarial drug candidates against this novel class of essential parasite enzymes.


An automatic step adjustment method for average power analysis technique used in fiber amplifiers.
An automatic step adjustment (ASA) method for average power analysis (APA) technique used in fiber amplifiers is proposed in this paper for the first time. In comparison with the traditional APA technique, the proposed method has suggested two unique merits such as a higher order accuracy and an ASA mechanism, so that it can significantly shorten the computing time and improve the solution accuracy. A test example demonstrates that, by comparing to the APA technique, the proposed method increases the computing speed by more than a hundredfold under the same errors. By computing the model equations of erbium-doped fiber amplifiers, the numerical results show that our method can improve the solution accuracy by over two orders of magnitude at the same amplifying section number. The proposed method has the capacity to rapidly and effectively compute the model equations of fiber Raman amplifiers and semiconductor lasers.


Geographical variation of overweight, obesity and related risk factors: Findings from the European Health Examination Survey in Luxembourg, 2013-2015.
The analyses of geographic variations in the prevalence of major chronic conditions, such as overweight and obesity, are an important public health tool to identify "hot spots" and inform allocation of funding for policy and health promotion campaigns, yet rarely performed. Here we aimed at exploring, for the first time in Luxembourg, potential geographic patterns in overweight/obesity prevalence in the country, adjusted for several demographic, socioeconomic, behavioural and health status characteristics. Data came from 720 men and 764 women, 25-64 years old, who participated in the European Health Examination Survey in Luxembourg (2013-2015). To investigate the geographical variation, geo-additive semi-parametric mixed model and Bayesian modelisations based on Markov Chain Monte Carlo techniques for inference were performed. Large disparities in the prevalence of overweight and obesity were found between municipalities, with the highest rates of obesity found in 3 municipalities located in the South-West of the country. Bayesian approach also underlined a nonlinear effect of age on overweight and obesity in both genders (significant in men) and highlighted the following risk factors: 1. country of birth for overweight in men born in a non-European country (Posterior Odds Ratio (POR): 3.24 [1.61-8.69]) and women born in Portugal (POR: 2.44 [1.25-4.43]), 2. low educational level (secondary or below) for overweight (POR: 1.66 (1.06-2.72)] and obesity (POR:2.09 [1.05-3.65]) in men, 3. single marital status for obesity in women (POR: 2.20 [1.24-3.91]), 4.fair (men: POR: 3.19 [1.58-6.79], women: POR: 2.24 [1.33-3.73]) to very bad health perception (men: POR: 15.01 [2.16-98.09]) for obesity, 5. sleeping more than 6 hours for obesity in unemployed men (POR: 3.66 [2.02-8.03]). Protective factors highlighted were: 1. single marital status against overweight (POR: [0.60 (0.38-0.96)]) and obesity (POR: 0.39 [0.16-0.84]) in men, 2. the fact to be widowed against overweight in women (POR: [0.30 (0.07-0.86)], as well as a non European country of birth (POR: 0.49 [0.19-0.98]), tertiary level of education (POR: 0.34 [0.18-0.64]), moderate alcohol consumption (POR: 0.54 [0.36-0.90]) and aerobic physical activity practice (POR: 0.44 [0.27-0.77]) against obesity in women. A double burden of environmental exposure due to historic mining and industrial activities and past economic vulnaribility in the South-West of the country may have participated to the higher prevalence of obesity found in this region. Other demographic, socioeconomic, behavioural and health status covariates could have been involved as well.


The effectiveness of e-learning in pediatric medical student education.
Electronic learning allows individualized education and may improve student performance. This study assessed the impact of e-modules about infection control and congenital infections on medical knowledge. A descriptive study was conducted involving third-year medical students on pediatric clerkship. e-Module content in three different formats was developed: a text monograph, a PowerPoint presentation, and a narrated PowerPoint lecture. Students' use of the e-modules was tracked, as was participation in the infectious disease rotation and the order of pediatric rotation. Pre- and posttests specific to the e-module content and National Board of Medical Examiners (NBME) pediatric exam scores were recorded. Among 67 participants, 63% of them visited at least one e-module. Neither accessing any e-modules, timing of pediatric clerkship, nor assignment to ID rotation resulted in improved posttest nor NBME scores. Seventy percent of students rated the e-modules as satisfactory and reported usage improved their confidence with the congenital infections topic. e-Modules did not improve student performance on NBME or posttest; however, they were perceived as satisfactory and to have improved confidence among those who used them. This study underscores the importance of formally evaluating electronic and other innovative curricula when implemented within existing medical education frameworks.


Paleopleurocapsa wopfnerii gen. et sp. nov.: A Late Precambrian alga and its modern counterpart.
Silicified dolomite of the approximately one billion year old Skillogalee Dolomite of the Adelaide Geosyncline, South Australia, contains organically preserved microfossils of a structurally complex, crustose pleurocapsalean cyanophyte, herein described as Paleopleurocapsa wopfnerii. Although actual cell contents have been degraded, lamellar sheath material faithfully preserves the morphology of the alga. Comparison with specimens of the modern genus Pleurocapsa Thuret demonstrates affinities at the family level and quite possibly even generic identity.


Factors associated with the annual change in forced expiratory volume in 1 second of officially acknowledged victims of pollution-related illness in improved environments: a longitudinal study.
[Purpose] We examined factors affecting annual change in pulmonary function in residents previously exposed to air pollution in an area where pollution has been reduced and a long time period has elapsed. [Subjects and Methods] Data of 730 officially acknowledged victims of pollution-related illness from an annual survey during 2000 to 2009 were analyzed. The primary outcome was forced expiratory volume in 1 second (FEV1), along with factors such as age, body composition, smoking habits, respiratory symptoms, and classification of medical management (an index of the need for treatment). Multiple regression analyses were used to identify factors associated with the annual change in FEV1. [Results] Three significant factors were identified: smoking habit, classification of medical management, and gender. Smoking habits and classification of medical management had stronger effects on the annual change in FEV1 than gender. [Conclusion] With an improved environment, continuation of smoking accelerates the decline in FEV1.


Statin therapy in kidney disease populations: potential benefits beyond lipid lowering and the need for clinical trials.
This review outlines the limited information currently available on the effects of statins among patients with chronic kidney disease, and summarizes the ongoing randomized trials designed to address this question. The effects of fluvastatin on major coronary events among renal transplant patients, and the effects of pravastatin and simvastatin in small subgroups of coronary patients with minor degrees of renal impairment, appear broadly compatible with those observed in trials conducted in non-renal populations. In addition, recent evidence from trials among patients with vascular disease or diabetes suggests that statin therapy may delay progressive loss of renal function. However, there remains substantial uncertainty regarding the effects of statin therapy among patients with established chronic kidney disease (pre-dialysis or dialysis patients). In particular, there does not appear to be a strongly positive relationship between blood cholesterol and cardiovascular events in such patients. This may be because uraemic cardiomyopathy and arteriosclerosis, which cause the majority of these events in chronic kidney disease patients, do not depend strongly on blood cholesterol. Statins appear effective for the prevention of vascular events in people with established vascular disease and mild renal impairment, and may delay renal disease progression in such individuals. However, the effects of statins in patients with established chronic kidney disease are unknown, and we await the results of ongoing large-scale randomized trials of statin therapy among such patients.


Selective attention supports working memory maintenance by modulating perceptual processing of distractors.
Selective attention has been shown to bias sensory processing in favor of relevant stimuli and against irrelevant or distracting stimuli in perceptual tasks. Increasing evidence suggests that selective attention plays an important role during working memory maintenance, possibly by biasing sensory processing in favor of to-be-remembered items. In the current study, we investigated whether selective attention may also support working memory by biasing processing against irrelevant and potentially distracting information. Event-related potentials (ERPs) were recorded while subjects (n = 22) performed a delayed-recognition task for faces and shoes. The delay period was filled with face or shoe distractors. Behavioral performance was impaired when distractors were congruent with the working memory domain (e.g., face distractor during working memory for faces) relative to when distractors were incongruent with the working memory domain (e.g., face distractor during shoe working memory). If attentional biasing against distractor processing is indeed functionally relevant in supporting working memory maintenance, perceptual processing of distractors is predicted to be attenuated when distractors are more behaviorally intrusive relative to when they are nonintrusive. As such, we predicted that perceptual processing of distracting faces, as measured by the face-sensitive N170 ERP component, would be reduced in the context of congruent (face) working memory relative to incongruent (shoe) working memory. The N170 elicited by distracting faces demonstrated reduced amplitude during congruent versus incongruent working memory. These results suggest that perceptual processing of distracting faces may be attenuated due to attentional biasing against sensory processing of distractors that are most behaviorally intrusive during working memory maintenance.


Beta-Arrestins: new roles in regulating heptahelical receptors' functions.
The last few years have seen a marked expansion in appreciation of the diversity of roles played by the betaArrestins in regulating GPCR functions. Originally discovered as molecules that desensitize such receptors, the roles of betaArrestins have expanded to include acting as signalling adapters or intermediates that recruit other key molecules to the GPCRs in an agonist-regulated fashion. For example, interactions with components of the endocytic machinery, such as clathrin, the adapter protein AP-2 and the N-ethylmaleimide sensitive fusion protein (NSF), demonstrate the ability of betaArrestins to act as adapters to facilitate the clathrin-mediated endocytosis of certain members of the GPCR family. BetaArrestins have also been shown to serve as signalling molecules. The Ras-dependent activation of ERK1/2 may involve the betaArrestin-dependent recruitment of c-Src to the beta2-adrenergic receptor (beta2-AR). More recently, betaArrestins have been shown to act as molecular scaffolds that coordinate the assembly of certain MAP kinase complexes that lead to the stimulation of either ERK1/2 or JNK3. Finally, long-term accumulation of arrestin-rhodopsin complexes, in photoreceptor cells has been shown to trigger apoptosis.


Light-trap design using multiple reflections and solid-angle attenuation: application to a spaceborne electron spectrometer.
The design and performance of a new light trap for a spaceborne electron spectrometer are described. The light trap has a measured photon-rejection ratio of 2 x 10(-11), allowing only one in 5 x 10(10) incident photons to reach the sensitive area of the instrument. This rejection is more than sufficient because the ambient ultraviolet in Earth orbit requires a rejection no better than 10(-8) to maintain the photon interference to less than 10 count/s. The light trap uses triple reflections to keep most of the light passing through the entrance slit away from the sensitive area of the spectrometer. However, because of electron-optic requirements, the edge of one of the metallic electrodes falls within the field of view of the sensitive area, allowing double-reflection photon paths to reach the sensitive area. Assuming diffuse reflectance r, the author shows that the photon rejection can be written as epsilon = G(2)r(2) + G(3)r(3) with G(3) approximately 10 times larger than G(2). Both coefficients depend only on the internal surface geometry; G(2) represents the electrode edge reflections (second-order) and G(3) represents the triple-reflection (third-order) paths. As shown by the analysis and measurements taken at two different values of r, the rejection is controlled by triple reflections if r > 0.08. It is shown that the average reflectance of all the internal surfaces must be less than 0.006, which is consistent with the data on the black coating applied to all surfaces. The analysis makes it possible to compare the photon contributions of each of the internal reflecting areas and to estimate the effective scattering width of the metallic electrode edge.


Efficacy of pharmacotherapies for short-term smoking abstinance: a systematic review and meta-analysis.
Smoking cessation has important immediate health benefits. The comparative short-term effectiveness of smoking cessation interventions is not well known. We aimed to determine the relative effectiveness of nicotine replacement therapy (NRT), bupropion and varenicline at 4 weeks post-target quit date. We searched 10 electronic medical databases (inception to October 2008). We selected randomized clinical trials [RCTs] evaluating interventions for our primary outcome of abstinence from smoking at at-least 4 weeks post-target quit date, with biochemical confirmation. We conducted random-effects odds ratio (OR) meta-analysis and meta-regression. We compared treatment effects across interventions using head-to-head trials and calculated indirect comparisons. We combined a total of 101 trials evaluating delivery of NRT versus inert controls at approximately 4 weeks post-target quit date (total n = 31,321). The pooled overall OR is OR 2.05 (95% Confidence Interval [CI], 1.89-2.23, P =< 0.0001). We pooled data from 31 bupropion trials contributing a total n of 11,118 participants and found a pooled OR of 2.25 (95% CI, 1.94-2.62, P =< 0.0001). We evaluated 9 varenicline trials compared to placebo. Our pooled estimate for cessation at 4 weeks post-target quit date found a pooled OR of 3.16 (95% CI, 2.55-3.91, P =< 0.0001). Two trials evaluated head to head comparisons of varenicline and bupropion and found a pooled estimate of OR 1.86 (95% CI, 1.49-2.33, P =< 0.0001 at 4 weeks post-target quit date. Indirect comparisons were: NRT and bupropion, OR, 1.09, 95% CI, 0.93-1.31, P = 0.28; varenicline and NRT, OR 1.56, 95% CI, 1.23-1.96, P = 0.0002; and, varenicline and bupropion, OR 1.40, 95% CI, 1.08-1.85, P = 0.01. Pharmacotherapeutic interventions are effective for increasing smoking abstinence rates in the short-term.


The overexpressed human 46-kDa mannose 6-phosphate receptor mediates endocytosis and sorting of beta-glucuronidase.
We studied the function of the human small (46-kDa) mannose 6-phosphate receptor (SMPR) in transfected mouse L cells that do not express the larger insulin-like growth factor II/mannose 6-phosphate receptor. Cells overexpressing human SMPR were studied for enzyme binding to cell surface receptors, for binding to intracellular receptors in permeabilized cells, and for receptor-mediated endocytosis of recombinant human beta-glucuronidase. Specific binding to human SMPR in permeabilized cells showed a pH optimum between pH 6.0 and pH 6.5. Binding was significant in the presence of EDTA but was enhanced by added divalent cations. Up to 2.3% of the total functional receptor could be detected on the cell surface by enzyme binding. We present experiments showing that at very high levels of overexpression, and at pH 6.5, human SMPR mediated the endocytosis of beta-glucuronidase. At pH 7.5, the rate of endocytosis was only 14% the rate seen at pH 6.5. Cells overexpressing human SMPR also showed reduced secretion of newly synthesized beta-glucuronidase when compared to cells transfected with vector only, suggesting that overexpressed human SMPR can participate in sorting of newly synthesized beta-glucuronidase and partially correct the sorting defect in mouse L cells that do not express the insulin-like growth factor II/mannose 6-phosphate receptor.


PERP regulates enamel formation via effects on cell-cell adhesion and gene expression.
Little is known about the role of cell-cell adhesion in the development of mineralized tissues. Here we report that PERP, a tetraspan membrane protein essential for epithelial integrity, regulates enamel formation. PERP is necessary for proper cell attachment and gene expression during tooth development, and its expression is controlled by P63, a master regulator of stratified epithelial development. During enamel formation, PERP is localized to the interface between the enamel-producing ameloblasts and the stratum intermedium (SI), a layer of cells subjacent to the ameloblasts. Perp-null mice display dramatic enamel defects, which are caused, in part, by the detachment of ameloblasts from the SI. Microarray analysis comparing gene expression in teeth of wild-type and Perp-null mice identified several differentially expressed genes during enamel formation. Analysis of these genes in ameloblast-derived LS8 cells upon knockdown of PERP confirmed the role for PERP in the regulation of gene expression. Together, our data show that PERP is necessary for the integrity of the ameloblast-SI interface and that a lack of Perp causes downregulation of genes that are required for proper enamel formation.


Toxicology and Carcinogenesis Studies of Naphthalene (CAS No. 91-20-3) in B6C3F1 Mice (Inhalation Studies).
Naphthalene, a white, crystalline powder, is used as a moth repellent and in the manufacture of phthalic and anthranilic acids, naphthylamines, and synthetic resins. The 2-year studies were conducted by exposing groups of male and female B6C3F1 mice to naphthalene (>99% pure) vapor for 6 hours daily, 5 days per week, for 104 weeks. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 2-Year Studies: Groups of male and female mice were exposed to atmospheres containing 0 (75 mice per group), 10 (75 mice per group), or 30 ppm (150 mice per group) naphthalene. Mice from each group were included for 14-day hematology evaluations (male: 0 ppm, 5 animals; 10 ppm, 4; 30 ppm, 10; female: 0 ppm, 4; 10 ppm, 5; 30 ppm, 10). Mean body weights of exposed mice were slightly lower than those of controls throughout the studies. Survival of male control mice was significantly less than that of exposed mice; the lower survival was the result of wound trauma and secondary infections related to fighting among the group-housed mice (0 ppm, 26/70, 37%; 10 ppm, 52/69, 75%; 30 ppm, 118/133, 89%). Survival of exposed female mice was similar to that of controls (59/69, 86%; 57/65, 88%; 102/135, 76%). Neoplastic and Nonneoplastic Effects in the 2-Year Studies: No increase in tumor incidence related to naphthalene administration was observed in male mice. In females, the incidence of pulmonary alveolar/bronchiolar adenomas was significantly greater in the high-dose group than in the controls (5/69, 7%; 2/65, 3%; 28/135, 21%). One other high-dose female had an alveolar/bronchiolar carcinoma. The combined incidence of alveolar/ bronchiolar adenomas and carcinomas in the high-dose females was above those for control female B6C3F1 mice from NTP feed, water, and inhalation studies (91/1,166, 7.8%, range 0%-16%). These lung tumors were attributed to naphthalene exposure. Nonneoplastic lesions attributed to naphthalene exposure were observed in the nose and lungs of mice of both sexes. In the nose, naphthalene exposure was associated with an increase in the incidence and severity of chronic inflammation, metaplasia of the olfactory epithelium, and hyperplasia of respiratory epithelium. Chronic inflammation in the lung was associated with chemical exposure. Genetic Toxicology: Naphthalene was negative for the induction of gene mutations in Salmonella typhimurium strains TA100, TA1535, TA1537, and TA98 with and without exogenous metabolic activation (S9). In cytogenetic tests with Chinese hamster ovary cells, naphthalene induced sister chromatid exchanges with and without S9 activation. Exposure to naphthalene induced a significant increase in chromosomal aberrations in Chinese hamster ovary cells in the presence of S9. Conclusions: Under the conditions of these 2-year inhalation studies, there was no evidence of carcinogenic activity of naphthalene in male B6C3F1 mice exposed to 10 or 30 ppm. There was some evidence of carcinogenic activity of naphthalene in female B6C3F1 mice, based on increased incidences of pulmonary alveolar/ bronchiolar adenomas. In both male and female mice, naphthalene caused increased incidences and severity of chronic inflammation, metaplasia of the olfactory epithelium, and hyperplasia of the respiratory epithelium in the nose and chronic inflammation in the lungs. Synonyms: Naphthalin, Naphthene, Tar Camphor


Racial differences in microbleed prevalence in primary intracerebral hemorrhage.
Primary intracerebral hemorrhage is two to three times more common in many racial populations, including black patients. Previous studies have shown that microbleeds, identified on gradient echo MRI (GRE), are present in 50-80% of patients with primary ICH. The objective of this study was to compare, by race, the rates, risk factors, and topography of microbleeds in patients hospitalized for primary ICH. Patients diagnosed with primary ICH at two metropolitan stroke centers were included. Clinical and neuroimaging data were recorded for each patient. Analyses were performed to compare baseline characteristics as well as imaging findings by race. A total of 87 patients met inclusion criteria (42 black subjects, 45 white subjects). The black cohort was younger (p < 0.001), and had a greater rate of hypertension (p = 0.001), but not other vascular risk factors. Microbleeds were more prevalent in the black population, with 74% of blacks having one or more microbleeds compared to 42% of whites (p = 0.005). The black population also tended to have a greater frequency of microbleeds in multiple territories than the white population (38% vs 22%, p = 0.106). When adjusting for age, hypertension, and alcohol use, race was an independent predictor of microbleeds (OR 3.308, 95% CI 1.144-9.571, p = 0.027). These pilot data suggest that significant racial differences exist in the frequency and topography of microbleeds in patients with primary ICH. Microbleeds may be an important emerging imaging biomarker with the potential to provide insights into ICH pathophysiology, prognosis, and disease progression, as well as possible therapeutic strategies, particularly in medically underserved populations.