Heat shock protein 27 influences the anti-cancer effect of curcumin in colon cancer cells through ROS production and autophagy activation.
The problem of therapeutic resistance and chemotherapeutic efficacy is tricky and critical in the management of colorectal cancer (CRC). Curcumin is a promising anti-cancer agent. Heat shock protein 27 (HSP27) is correlated with CRC progression and is said to affect CRC response to different therapies. However, the role of HSP27 on the therapeutic efficacy of curcumin remains unknown. HSP27 was silenced using small hairpin RNA (shRNA) technique. The cytotoxic and apoptotic effects of curcumin were assessed by sulforhodamine B (SRB) colorimetric assay, flow cytometric cell cycle analysis, and annexin V/propidium iodide (PI) double-labeling assays. Total reactive oxygen species (ROS)/superoxide and autophagy detection were performed, and the levels of apoptosis-related proteins were examined by Western blotting. It was found that the silencing of HSP27 (HSP27-KD) resulted in increased treatment resistance to curcumin in CRC cells. In addition, cell cycle analysis showed that the curcumin treatment caused cell cycle arrest at the G2/M phase in the control group, and apoptosis was reduced in the HSP27-KD group. Curcumin treatment also resulted in a decrease in anti-apoptotic proteins, p-Akt, Akt, Bcl-2 and p-Bad, and increase in pro-apoptotic proteins Bad and c-PARP levels in the control cells but not in the HSP27-KD cells. This was also followed by low reactive oxygen/nitrogen species (ROS/RNS), superoxide and autophagy induction levels in the HSP27-KD cells as compared to the control cells. Therefore, as silencing of HSP27 increases curcumin resistance by reducing apoptosis and reactive oxidative stress production, HSP27 is a potential selective target for curcumin treatment in CRC.


First Report of Tomato spotted wilt virus on Brugmansia sp. in Serbia.
Brugmansia (Brugmansia spp.), also known as Angel's trumpet, is a perennial shrub in the Solanaceae that is a popular landscape plant in the tropics and subtropics, and potted plant in temperate regions. In April 2012, virus-like symptoms including chlorotic leaf patterns and curling followed by necrosis and distortion of leaves were observed on five outdoor-grown brugmansia plants in a private garden in Mackovac, Rasina District, Serbia. Symptomatic leaves were tested for the presence of several common ornamental viruses including Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), Cucumber mosaic virus (CMV), and Tobacco mosaic virus (TMV) by commercial double-antibody sandwich (DAS)-ELISA diagnostic kits (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls and extract from healthy brugmansia leaves were included in each ELISA. TSWV was detected serologically in all five brugmansia samples and all tested samples were negative for INSV, CMV, and TMV. The virus was mechanically transmitted from an ELISA-positive sample (41-12) to five plants of each Petuina × hybrida and Nicotiana glutinosa. Inoculated P. × hybrida plants showed local necrotic lesions and N. glutinosa showed mosaic and systemic necrosis 4 and 12 days post-inoculation, respectively, which were consistent with symptoms caused by TSWV (1). For further confirmation of TSWV infection, reverse transcription (RT)-PCR was performed with the OneStep RT-PCR (Qiagen, Hilden, Germany) using a set of TSWV-specific primers, TSWV CP-f and TSWV CP-r (4), designed to amplify a 738-bp fragment of the nucleocapsid protein (N) gene. Total RNAs from naturally infected brugmansia and symptomatic N. glutinosa plants were extracted using the RNeasy Plant Mini Kit (Qiagen). Total RNAs obtained from the Serbian tobacco isolate of TSWV (GenBank Accession No. GQ373173) and healthy brugmansia plants were used as positive and negative controls, respectively. The expected size of the RT-PCR product was amplified from symptomatic brugmansia and N. glutinosa but not from healthy tissues. The amplified product derived from the isolate 41-12 was sequenced directly after purification with the QIAquick PCR Purification kit (Qiagen), deposited in GenBank (JX468080), and subjected to sequence analysis by MEGA5 software (3). Sequence comparisons revealed that the Serbian isolate 41-12 shared the highest nucleotide identity of 99.9% (99.5% amino acid identity) with an Italian TSWV isolate P105/2006RB (DQ915946) originating from pepper. To our knowledge, this is the first report of TSWV on brugmansia in Serbia. Due to the increasing popularity and economic importance of brugmansia as an ornamental crop, thorough inspections and subsequent testing for TSWV and other viruses are needed. This high-value ornamental plant may act also as reservoir for the virus that can infect other ornamentals and cultivated crops, considering that TSWV has a very broad host range (2). References: (1) Anonymous. OEPP/EPPO Bull. 34:271, 2004. (2) G. Parrella et al. J. Plant Pathol. 85:227, 2003. (3) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011. (4) A. Vučurović et al. Eur. J. Plant Pathol. 133:935, 2012.


Mu-8: visualizing differences between proteins and their families.
A complete understanding of the relationship between the amino acid sequence and resulting protein function remains an open problem in the biophysical sciences. Current approaches often rely on diagnosing functionally relevant mutations by determining whether an amino acid frequently occurs at a specific position within the protein family. However, these methods do not account for the biophysical properties and the 3D structure of the protein. We have developed an interactive visualization technique, Mu-8, that provides researchers with a holistic view of the differences of a selected protein with respect to a family of homologous proteins. Mu-8 helps to identify areas of the protein that exhibit: (1) significantly different bio-chemical characteristics, (2) relative conservation in the family, and (3) proximity to other regions that have suspect behavior in the folded protein. Our approach quantifies and communicates the difference between a reference protein and its family based on amino acid indices or principal components of amino acid index classes, while accounting for conservation, proximity amongst residues, and overall 3D structure. We demonstrate Mu-8 in a case study with data provided by the 2013 BioVis contest. When comparing the sequence of a dysfunctional protein to its functional family, Mu-8 reveals several candidate regions that may cause function to break down.


Predicting Optimal Outcomes in Cognitive Therapy or Interpersonal Psychotherapy for Depressed Individuals Using the Personalized Advantage Index Approach.
Although psychotherapies for depression produce equivalent outcomes, individual patients respond differently to different therapies. Predictors of outcome have been identified in the context of randomized trials, but this information has not been used to predict which treatment works best for the depressed individual. In this paper, we aim to replicate a recently developed treatment selection method, using data from an RCT comparing the effects of cognitive therapy (CT) and interpersonal psychotherapy (IPT). 134 depressed patients completed the pre- and post-treatment BDI-II assessment. First, we identified baseline predictors and moderators. Second, individual treatment recommendations were generated by combining the identified predictors and moderators in an algorithm that produces the Personalized Advantage Index (PAI), a measure of the predicted advantage in one therapy compared to the other, using standard regression analyses and the leave-one-out cross-validation approach. We found five predictors (gender, employment status, anxiety, personality disorder and quality of life) and six moderators (somatic complaints, cognitive problems, paranoid symptoms, interpersonal self-sacrificing, attributional style and number of life events) of treatment outcome. The mean average PAI value was 8.9 BDI points, and 63% of the sample was predicted to have a clinically meaningful advantage in one of the therapies. Those who were randomized to their predicted optimal treatment (either CT or IPT) had an observed mean end-BDI of 11.8, while those who received their predicted non-optimal treatment had an end-BDI of 17.8 (effect size for the difference = 0.51). Depressed patients who were randomized to their predicted optimal treatment fared much better than those randomized to their predicted non-optimal treatment. The PAI provides a great opportunity for formal decision-making to improve individual patient outcomes in depression. Although the utility of the PAI approach will need to be evaluated in prospective research, this study promotes the development of a treatment selection approach that can be used in regular mental health care, advancing the goals of personalized medicine.


Should the provision of home help services be contained?: validation of the new preventive care policy in Japan.
To maintain the sustainability of public long-term care insurance (LTCI) in Japan, a preventive care policy was introduced in 2006 that seeks to promote active improvement in functional status of elderly people who need only light care. This policy promotes the use of day care services to facilitate functional improvement, and contains the use of home help services that provide instrumental activity of daily living (IADL) support. However, the validity of this approach remains to be demonstrated. Subjects comprised 241 people aged 65 years and over who had recently been certified as being eligible for the lightest eligibility level and had began using either home help or day care services between April 2007 and October 2008 in a suburban city of Tokyo. A retrospective cohort study was conducted ending October 2009 to assess changes in the LTCI eligibility level of these subjects. Cox's proportional hazards model was used to calculate the relative risk of declining in function to eligibility Level 4 among users of the respective services. Multivariate analysis adjusted for factors related to service use demonstrated that the risk of decline in functional status was lower for users of home help services than for users of day care services (HR = 0.55, 95% CI: 0.31-0.98). The same result was obtained when stratified by whether the subject lived with family or not. Furthermore, those who used two or more hours of home help services did not show an increase in risk of decline when compared with those who used less than two hours. No evidence was obtained to support the effectiveness of the policy of promoting day care services and containing home help services for those requiring light care.


Exceptional lability of a genomic complex in rice and its close relatives revealed by interspecific and intraspecific comparison and population analysis.
Extensive DNA rearrangement of genic colinearity, as revealed by comparison of orthologous genomic regions, has been shown to be a general concept describing evolutionary dynamics of plant genomes. However, the nature, timing, lineages and adaptation of local genomic rearrangement in closely related species (e.g., within a genus) and haplotype variation of genomic rearrangement within populations have not been well documented. We previously identified a hotspot for genic rearrangement and transposon accumulation in the Orp region of Asian rice (Oryza sativa, AA) by comparison with its orthologous region in sorghum. Here, we report the comparative analysis of this region with its orthologous regions in the wild progenitor species (O. nivara, AA) of Asian rice and African rice (O. glaberrima) using the BB genome Oryza species (O. punctata) as an outgroup, and investigation of transposon insertion sites and a segmental inversion event in the AA genomes at the population level. We found that Orp region was primarily and recently expanded in the Asian rice species O. sativa and O. nivara. LTR-retrotransposons shared by the three AA-genomic regions have been fixed in all the 94 varieties that represent different populations of the AA-genome species/subspecies, indicating their adaptive role in genome differentiation. However, LTR-retrotransposons unique to either O. nivara or O. sativa regions exhibited dramatic haplotype variation regarding their presence or absence between or within populations/subpopulations. The LTR-retrotransposon insertion hotspot in the Orp region was formed recently, independently and concurrently in different AA-genome species, and that the genic rearrangements detected in different species appear to be differentially triggered by transposable elements. This region is located near the end of the short arm of chromosome 8 and contains a high proportion of LTR-retrotransposons similar to observed in the centromeric region of this same chromosome, and thus may represent a genomic region that has recently switched from euchromatic to heterochromatic states. The haplotype variation of LTR-retrotransposon insertions within this region reveals substantial admixture among various subpopulations as established by molecular markers at the whole genome level, and can be used to develop retrotransposon junction markers for simple and rapid classification of O. sativa germplasm.


Influence of the crown-to-implant length ratio on the clinical performance of implants supporting single crown restorations: a cross-sectional retrospective 5-year investigation.
The aim of this study was to investigate the influence of the crown-to-implant length ratio (c/i ratio) on the implant survival, changes of the marginal bone level (MBL) and the occurrence of biological and technical complications. This cross-sectional retrospective study included all patients with implants in the posterior segments supporting single crown restorations with a minimum follow-up of 5 years. All patients were questioned and examined clinically and radiographically. The technical and biological c/i ratio and the MBL were measured on digitized periapical radiographs. The following outcome parameters in relation to the c/i ratio and the co-factors were statistically analyzed: implant survival rate, MBL, occurrence of technical and biological complications. For statistical analysis, regression, correlation and survival analyses were applied (P<0.05). Seventy patients (mean age of 50.7 years [range 19.8-76.6 years]) with a total of 100 implants (24 Straumann type, 76 Brånemark type) were included in this study. The mean follow-up period was 6.2 years (range 4.73-11.7 years). Six implants failed during the follow-up period, yielding a cumulative survival rate of 95.8% at 5 years in function. The mean technical c/i ratio was 1.04 (±0.26, range 0.59-2.01). The mean biological c/i ratio was 1.48 (±0.42, range 0.82-3.24). No statistically significant influence of the technical and biological c/i ratio was found on the implant survival, MBL and occurrence of technical and biological complications. When adjusted for the biological c/i ratio, smoking was the only co-factor significantly associated with implant failure and biological complications. In the present study, the c/i ratio did not influence the clinical performance of implants supporting single crown restorations in the posterior segments of the jaw within the range tested.


Assessing bird avoidance of high-contrast lights using a choice test approach: implications for reducing human-induced avian mortality.
Avian collisions with man-made objects and vehicles (e.g., buildings, cars, airplanes, power lines) have increased recently. Lights have been proposed to alert birds and minimize the chances of collisions, but it is challenging to choose lights that are tuned to the avian eye and can also lead to avoidance given the differences between human and avian vision. We propose a choice test to address this problem by first identifying wavelengths of light that would over-stimulate the retina using species-specific perceptual models and by then assessing the avoidance/attraction responses of brown-headed cowbirds to these lights during daytime using a behavioral assay. We used perceptual models to estimate wavelength-specific light emitting diode (LED) lights with high chromatic contrast. The behavioral assay consisted of an arena where the bird moved in a single direction and was forced to make a choice (right/left) using a single-choice design (one side with the light on, the other with the light off) under diurnal light conditions. First, we identified lights with high saliency from the cowbird visual perspective: LED lights with peaks at 380 nm (ultraviolet), 470 nm (blue), 525 nm (green), 630 nm (red), and broad-spectrum (white) LED lights. Second, we found that cowbirds significantly avoided LED lights with peaks at 470 and 630 nm, but did not avoid or prefer LED lights with peaks at 380 and 525 nm or white lights. The two lights avoided had the highest chromatic contrast but relatively lower levels of achromatic contrast. Our approach can optimize limited resources to narrow down wavelengths of light with high visual saliency for a target species leading to avoidance. These lights can be used as candidates for visual deterrents to reduce collisions with man-made objects and vehicles.


Methodological approaches for monitoring opportunistic pathogens in premise plumbing: A review.
Opportunistic premise (i.e., building) plumbing pathogens (OPPPs, e.g., Legionella pneumophila, Mycobacterium avium complex, Pseudomonas aeruginosa, Acanthamoeba, and Naegleria fowleri) are a significant and growing source of disease. Because OPPPs establish and grow as part of the native drinking water microbiota, they do not correspond to fecal indicators, presenting a major challenge to standard drinking water monitoring practices. Further, different OPPPs present distinct requirements for sampling, preservation, and analysis, creating an impediment to their parallel detection. The aim of this critical review is to evaluate the state of the science of monitoring OPPPs and identify a path forward for their parallel detection and quantification in a manner commensurate with the need for reliable data that is informative to risk assessment and mitigation. Water and biofilm sampling procedures, as well as factors influencing sample representativeness and detection sensitivity, are critically evaluated with respect to the five representative bacterial and amoebal OPPPs noted above. Available culturing and molecular approaches are discussed in terms of their advantages, limitations, and applicability. Knowledge gaps and research needs towards standardized approaches are identified.


Use of health information technology to manage frequently presenting emergency department patients.
To determine if the effective use of Health Information Technologies (HIT) and the Electronic Medical Record (EMR) affects emergency department (ED) usage in a complicated frequently presenting patient population. A retrospective, observational study of 45 patients enrolled in our Frequent User Program called Community Resources for Emergency Department Overuse (CREDO) between June 2005 and July 2007. The study was conducted at an urban hospital with greater than 95,000 annual visits. Patients served as their own historical controls. In this pre-post study, the pre-intervention control period was determined by the number of months the patient had been enrolled in the program. The pre- and post-intervention time periods were the same for each patient but varied between patients. The intervention included using HIT to identify the most frequently presenting patients and creating individualized care plans for those patients. The care plans were made available through the EMR to all healthcare providers. Study variables in this study intervention included ED charges, lab studies ordered, number of ED visits, length of stay (LOS), and Total Emergency Department Contact Time (TEDCT), which is the product of the number of visits and the LOS. We analyzed these variables using paired T-tests. This study was approved by the institutional review board. Forty-five patients were enrolled, but nine were excluded for no post enrollment visits; thus, statistical analysis was conducted with n=36. The ED charges decreased by 24% from $64,721 to $49,208 (p=0.049). The number of lab studies ordered decreased by 28% from 1847 to 1328 (p=0.04). The average number of ED visits/patient decreased by 25% from 67.4 to 50.5 (p=0.046). The TEDCT decreased by 39% from 443.7 hours to 270.6 hours (p=0.003). In this pre-post analysis of an intervention targeting ED frequent users, the use of HIT and the EMR to identify patients and store easily accessible care plans significantly reduced ED charges, labs ordered, number of ED visits, and the TEDCT.


Effects of n-3 fatty acids on cognitive decline: a randomized, double-blind, placebo-controlled trial in stable myocardial infarction patients.
Epidemiological studies suggest a protective effect of n-3 fatty acids derived from fish (eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) against cognitive decline. For α-linolenic acid (ALA) obtained from vegetable sources, the effect on cognitive decline is unknown. We examined the effect of n-3 fatty acid supplementation on cognitive decline in coronary heart disease patients. The analysis included 2911 coronary patients (78% men) aged 60 to 80 years who participated in a double-blind placebo-controlled trial of n-3 fatty acids and cardiovascular diseases (Alpha Omega Trial). By using a 2 × 2 factorial design, patients were randomly assigned to margarines that provided 400 mg/d of EPA-DHA, 2 g/d of ALA, both EPA-DHA and ALA, or placebo for 40 months. Cognitive function was assessed by the Mini-Mental State Examination (MMSE) at baseline and after 40 months. The effect of n-3 fatty acids on change in MMSE score was assessed using analysis of variance. Logistic regression analysis was used to examine the effects on risk of cognitive decline, defined as a decrease of 3 or more points in MMSE score or incidence of dementia. Patients in the active treatment groups had an additional intake of 384 mg of EPA-DHA, 1.9 g of ALA, or both. The overall MMSE score in this cohort was 28.3 ± 1.6 points, which decreased by 0.67 ± 2.25 points during follow-up. Changes in MMSE score during intervention did not differ significantly between EPA-DHA and placebo (-0.65 vs -0.69 points, P = .44) or between ALA and placebo (-0.60 vs -0.74 points, P = .12). The risk of cognitive decline was 1.03 (95% confidence interval: 0.84-1.26, P = .80) for EPA-DHA (vs placebo) and 0.90 (0.74-1.10, P = .31) for ALA (vs placebo). This large intervention study showed no effect of dietary doses of n-3 fatty acids on global cognitive decline in coronary heart disease patients.


Gamma Knife radiosurgery for larger-volume vestibular schwannomas. Clinical article.
Stereotactic radiosurgery (SRS) is an important management option for patients with small- and medium-sized vestibular schwannomas. To assess the potential role of SRS in larger tumors, the authors reviewed their recent experience. Between 1994 and 2008, 65 patients with vestibular schwannomas between 3 and 4 cm in one extracanalicular maximum diameter (median tumor volume 9 ml) underwent Gamma Knife surgery. Seventeen patients (26%) had previously undergone resection. The median follow-up duration was 36 months (range 1-146 months). At the first planned imaging follow-up at 6 months, 5 tumors (8%) were slightly expanded, 53 (82%) were stable in size, and 7 (11%) were smaller. Two patients (3%) underwent resection within 6 months due to progressive symptoms. Two years later, with 63 tumors overall after the 2 post-SRS resections, 16 tumors (25%) had a volume reduction of more than 50%, 22 (35%) tumors had a volume reduction of 10-50%, 18 (29%) were stable in volume (volume change < 10%), and 7 (11%) had larger volumes (5 of the 7 patients underwent resection and 1 of the 7 underwent repeat SRS). Eighteen (82%) of 22 patients with serviceable hearing before SRS still had serviceable hearing after SRS more than 2 years later. Three patients (5%) developed symptomatic hydrocephalus and underwent placement of a ventriculoperitoneal shunt. In 4 patients (6%) trigeminal sensory dysfunction developed, and in 1 patient (2%) mild facial weakness (House-Brackmann Grade II) developed after SRS. In univariate analysis, patients who had a previous resection (p = 0.010), those with a tumor volume exceeding 10 ml (p = 0.05), and those with Koos Grade 4 tumors (p = 0.02) had less likelihood of tumor control after SRS. Although microsurgical resection remains the primary management choice in patients with low comorbidities, most vestibular schwannomas with a maximum diameter less than 4 cm and without significant mass effect can be managed satisfactorily with Gamma Knife radiosurgery.


Genome-wide epistatic interaction analysis reveals complex genetic determinants of circadian behavior in mice.
Genetic heterogeneity underlies many phenotypic variations observed in circadian rhythmicity. Continuous distributions in measures of circadian behavior observed among multiple inbred strains of mice suggest that the inherent contributions to variability are polygenic in nature. To identify genetic loci that underlie this complex behavior, we have carried out a genome-wide complex trait analysis in 196 (C57BL/6J X BALB/cJ)F(2) hybrid mice. We have characterized variation in this panel of F(2) mice among five circadian phenotypes: free-running circadian period, phase angle of entrainment, amplitude of the circadian rhythm, circadian activity level, and dissociation of rhythmicity. Our genetic analyses of these phenotypes have led to the identification of 14 loci having significant effects on this behavior, including significant main effect loci that contribute to three of these phenotypic measures: period, phase, and amplitude. We describe an additional locus detection method, genome-wide genetic interaction analysis, developed to identify locus pairs that may interact epistatically to significantly affect phenotype. Using this analysis, we identified two additional pairs of loci that have significant effects on dissociation and activity level; we also detected interaction effects in loci contributing to differences of period, phase, and amplitude. Although single gene mutations can affect circadian rhythms, the analysis of interstrain variants demonstrates that significant genetic complexity underlies this behavior. Importantly, most of the loci that we have detected by these methods map to locations that differ from the nine known clock genes, indicating the presence of additional clock-relevant genes in the mammalian circadian system. These data demonstrate the analytical value of both genome-wide complex trait and epistatic interaction analyses in further understanding complex phenotypes, and point to promising approaches for genetic analysis of such phenotypes in other mammals, including humans.


Therapeutic application of mesenchymal stem cells in bone and joint diseases.
Mesenchymal stem cells (MSCs), the non-hematopoietic progenitor cells, are multi-potent stem cells from a variety of tissues with the capability of self-renewal, proliferation, differentiation into multi-lineage cell types, as well as anti-inflammatory and immunomodulatory. These properties make MSCs an ideal source of cell therapy in bone and joint diseases. This review describes the advances of animal study and preliminary clinical application in the past few years, related to MSC-based cell therapy in the common bone and joint diseases, including osteoarthritis, rheumatoid arthritis, osteoporosis, osteonecrosis of the femoral head and osteogenesis imperfecta. It highlights the promising prospect of MSC in clinical application of bone and joint diseases.


Assessment of carbon in woody plants and soil across a vineyard-woodland landscape.
Quantification of ecosystem services, such as carbon (C) storage, can demonstrate the benefits of managing for both production and habitat conservation in agricultural landscapes. In this study, we evaluated C stocks and woody plant diversity across vineyard blocks and adjoining woodland ecosystems (wildlands) for an organic vineyard in northern California. Carbon was measured in soil from 44 one m deep pits, and in aboveground woody biomass from 93 vegetation plots. These data were combined with physical landscape variables to model C stocks using a geographic information system and multivariate linear regression. Field data showed wildlands to be heterogeneous in both C stocks and woody tree diversity, reflecting the mosaic of several different vegetation types, and storing on average 36.8 Mg C/ha in aboveground woody biomass and 89.3 Mg C/ha in soil. Not surprisingly, vineyard blocks showed less variation in above- and belowground C, with an average of 3.0 and 84.1 Mg C/ha, respectively. This research demonstrates that vineyards managed with practices that conserve some fraction of adjoining wildlands yield benefits for increasing overall C stocks and species and habitat diversity in integrated agricultural landscapes. For such complex landscapes, high resolution spatial modeling is challenging and requires accurate characterization of the landscape by vegetation type, physical structure, sufficient sampling, and allometric equations that relate tree species to each landscape. Geographic information systems and remote sensing techniques are useful for integrating the above variables into an analysis platform to estimate C stocks in these working landscapes, thereby helping land managers qualify for greenhouse gas mitigation credits. Carbon policy in California, however, shows a lack of focus on C stocks compared to emissions, and on agriculture compared to other sectors. Correcting these policy shortcomings could create incentives for ecosystem service provision, including C storage, as well as encourage better farm stewardship and habitat conservation.


Is a test of cure necessary following treatment for cervical infection with Chlamydia trachomatis?
Tests of cure (TOC) were performed on specimens from 106 women following treatment for chlamydial infection of the cervix. 91 women attended for the first TOC with enzyme immunoassay (EIA) within one week of finishing antibiotics. Three were EIA positive, 88 were EIA negative. These three women were subsequently EIA and culture negative although they received no further antibiotics. Ninety women returned for the second TOC with EIA and culture between seven and 27 days after completing treatment. All the results were negative. Routine TOC is unnecessary following appropriate antichlamydial therapy. If TOC is indicated antigen-detection methods should not be used immediately after finishing antibiotics as misleading positive results may be obtained. EIA or culture at one week or later after treatment is reliable.


Purinergic signaling involved in Müller cell function in the mammalian retina.
Purines (in particular, ATP and adenosine) act as neuro- and gliotransmitters in the sensory retina where they are involved in bidirectional neuron-glia signaling. This review summarizes the present knowledge about the expression and functional importance of P1 (adenosine) and P2 (nucleotide) receptors in Müller glial cells of the mammalian retina. Mammalian Müller cells express various subtypes of adenosine receptors and metabotropic P2Y receptors. Human Müller cells also express ionotropic P2X(7) receptors. Müller cells release ATP upon activation of metabotropic glutamate receptors and/or osmotic membrane stretching. The osmotic mechanism is abrogated under conditions associated with ischemia-hypoxia and inflammation, resulting in swelling of the Müller cells when the extracellular milieu is hypoosmotic. However, exogenous glutamate, which induces the release of ATP and adenosine, and thus activates P2Y(1) and A(1) adenosine receptors, respectively, prevents such osmotic swelling under pathological conditions, suggesting unimpaired receptor-induced release of ATP. In addition to the inhibition of swelling, which is implicated in regulating the volume of the extracellular space, purinergic signaling is involved in mediating neurovascular coupling. Furthermore, purinergic signals stimulate the proliferation of retinal precursor cells and Müller cells. In normal retinal information processing, Müller cells regulate the synaptic activity by the release of ATP and adenosine. In retinopathies, abrogation of the osmotic release of ATP, and the upregulation of ecto-apyrase (NTPDase1), may have neuroprotective effects by preventing the overactivation of neuronal P2X receptors that are implicated in apoptotic cell death. Pharmacological modulation of purinergic receptors of Müller cells may have clinical importance, e.g., for the clearance of retinal edema and for the inhibition of dysregulated cell proliferation in proliferative retinopathies.


Contribution of dose and frequency of administration to the therapeutic effect of growth hormone.
Forty two prepubertal children who were growth hormone insufficient were treated for the first time with 12 IU of biosynthetic methionyl growth hormone a week in three subcutaneous regimens for one year after one year of pretreatment observation. Thirteen received 4 IU growth hormone three days a week (group 1), 21 received 2 IU six days a week (group 2), and eight received 1 IU twice daily six days a week (group 3). The mean (SD) changes in height velocity standard deviation score (SDS) were +3.8 (1.7) in group 1, +5.3 (2.6) in group 2, and +5.9 (2.7) in group 3. There was a highly significant relation between the change in height velocity SDS and pretreatment height velocity SDS with a significant difference between the response of subjects receiving less than 15 IU/m2/week or greater than 15 IU/m2/week growth hormone. Age, bone age, height SDS, and height for bone age SDS did not contribute to the prediction of response during the first year of treatment. We recommend that growth hormone should be given in a dose of at least 15 IU/m2/week by equally divided daily subcutaneous injections.


Physical activity and other lifestyle behaviors in a Portuguese sample of adults: results from the Azorean Physical Activity and Health Study.
The aims of this study were to describe physical activity (PA) prevalence and compare it with other countries and to investigate possible associations between PA and other lifestyle behaviors in Azorean adults. 9991 adults (5723 women), aged 37.8 +/- 9.5 years, of the 2004 Azorean Physical Activity and Health Study. IPAQ assessed PA. All other lifestyle behaviors (age, gender, education level, income, employment, marital status, number of children, meal frequency, sleep time, sitting time, body mass index and alcohol and tobacco consumptions) were also self-reported. 57.1% of the participants met current PA recommendations and 32.2% were categorized as Health Enhancing PA (HEPA). Women were less likely to achieve PA recommendations, as well as the HEPA level. In both genders, higher education level, employment status, higher income, and sitting for more than 3h/day were negative predictors of HEPA; and, having at least 5 meals/day was positive predictor for the same PA level. There is a significant proportion of Azoreans, particularly women, that does not do enough PA. Targeted programs for Azoreans aimed to increase PA levels should pay special attention on women, and consider a multifactorial approach, once several lifestyle behaviors seem to interact with PA levels, in this population.


On interviewing people with pets: reflections from qualitative research on people with long-term conditions.
There is mounting evidence that pets are associated with physiological, psychological and social benefits for humans. Much of this research has come from western countries, where there have been consistent calls for greater engagement with pet ownership and health. Drawing on a secondary analysis of 61 in-depth interviews with people, or carers of people, with long-term conditions, we explore how pets feature in people's narrative accounts of their experiences. Our findings demonstrate the multifaceted nature of people's relationships with pets, and the embedded and embodied ways in which human-nonhuman interactions are played out in narratives of chronic illness. Our study differs from other work on pets and health in that, by returning to the interview video recordings, we were able to explore the sometimes three-way interactions, the co(a)gency, between participants, pets and researchers. Pets were often presented as important family members, yet the researchers' responses to the presence or talk about pets was often markedly different from their reactions to other household members. We conclude with cautioning against the downgrading of pets in narrative health research. Narrative approaches may invite participants to talk about what is important to them, yet they clearly have limitations in practice.


Assessment of nitrogen ceilings for Dutch agricultural soils to avoid adverse environmental impacts.
In the Netherlands, high traffic density and intensive animal husbandry have led to high emissions of reactive nitrogen (N) into the environment. This leads to a series of environmental impacts, including: (1) nitrate (NO3) contamination of drinking water, (2) eutrophication of freshwater lakes, (3) acidification and biodiversity impacts on terrestrial ecosystems, (4) ozone and particle formation affecting human health, and (5) global climate change induced by emissions of N2O. Measures to control reactive N emissions were, up to now, directed towards those different environmental themes. Here we summarize the results of a study to analyse the agricultural N problem in the Netherlands in an integrated way, which means that all relevant aspects are taken into account simultaneously. A simple N balance model was developed, representing all crucial processes in the N chain, to calculate acceptable N inputs to the farm (so-called N ceiling) and to the soil surface (application in the field) by feed concentrates, organic manure, fertiliser, deposition, and N fixation. The N ceilings were calculated on the basis of critical limits for NO 3 concentrations in groundwater, N concentrations in surface water, and ammonia (NH3) emission targets related to the protection of biodiversity of natural areas. Results show that in most parts of the Netherlands, except the western and the northern part, the N ceilings are limited by NH 3 emissions, which are derived from critical N loads for nature areas, rather than limits for both ground- and surface water. On the national scale, the N ceiling ranges between 372 and 858 kton year(-1) depending on the choice of critical limits. The current N import is 848 kton year(-1). A decrease of nearly 60% is needed to reach the ceilings that are necessary to protect the environment against all adverse impacts of N pollution from agriculture.


Spectrum of 68Ga-DOTA TATE Uptake in Patients With Neuroendocrine Tumors.
To analyze the biodistribution of Ga-DOTA-TATE in the normal tissues and uptake in benign, indeterminate, and malignant lesions in a population of patients with known neuroendocrine tumors (NET) using semiquantitative standardized uptake values (SUV) measurements. One hundred four consecutively scanned patients (51 men and 53 women; mean age, 56.4 years) with confirmed diagnosis of NET underwent PET/CT 1 hour after administration of Ga-DOTA-TATE. SUVmean, and SUVmax were measured in 37 normal anatomical structures for each patient. Abnormal uptake was divided into benign, indeterminate, and malignant categories based on imaging characteristic, clinical follow-up, and pathology. High physiologic uptake (SUVmax > 7) was observed in spleen, renal parenchyma, adrenal glands, pituitary gland, stomach, and liver (in decreasing order). Moderate uptake (3.5-7) was present in the prostate, jejunum, pancreas, ileum, and salivary glands. Mild uptake (2-3.5) was present in the uterus, colon, thyroid, rectum, and skeleton. A total of 678 lesions (limited to 5 lesions with highest uptake per organ) were included in the analysis, including 127 benign and 54 indeterminate lesions. Uptake was significantly higher in malignant lesions than in benign lesions, but an overlap was noted between the groups. Ga-DOTA TATE uptake in normal and abnormal structures is highly variable in patients with NET. SUV is a useful measure for characterizing benign versus malignant lesions. Anatomical and clinical correlation may be necessary to characterize foci of intermediate uptake.


To take responsibility or to be an onlooker. Nursing students' experiences of two models of supervision.
The present study aimed at describing how nursing students engaged in their clinical practice experienced two models of supervision: supervision on student wards and traditional supervision. Supervision for nursing students in clinical practice can be organized in different ways. In the present study, parts of nursing students' clinical practice were carried out on student wards in existing hospital departments. The purpose was to give students the opportunity to assume greater responsibility for their clinical education and to apply the nursing process more independently through peer learning. A descriptive design with a qualitative approach was used. Interviews were carried out with eight nursing students in their final semester of a 3-year degree program in nursing. The data were analyzed using content analysis. Two themes were revealed in the data analysis: When supervised on the student wards, nursing students experienced assuming responsibility and finding one's professional role, while during traditional supervision, they experienced being an onlooker and having difficulties assuming responsibility. Supervision on a student ward was found to give nursing students a feeling of acknowledgment and more opportunities to develop independence, continuity, cooperation and confidence.


Keratoprosthesis with biocolonizable microporous fluorocarbon haptic. Preliminary results in a 24-patient study.
Most complications of a keratoprosthesis occur at the tissue-to-implant interface. The ideal prosthesis would eliminate this interface by having the tissue actually grow into the supporting material. We present a prospective clinical human study of a novel biocolonizable keratoprosthesis in 24 eyes of 24 patients. To promote implant stability, the 9-mm-diameter haptic was fashioned using a custom-made microporous fluorocarbon with a 4-mm-diameter, 2.67-mm-long, central optic made of medical grade polymethylmethacrylate, giving a global visual field of 110 degrees to 130 degrees. Only bilaterally blind patients with untreatable corneal diseases were included in the study. The haptic was inserted into a lamellar pocket delaminated in the stroma, and the optic was positioned through a hole trephined in the central cornea. The average follow-up was 15.7 months (range, 4 to 28 months). The host corneal fibroblasts penetrated and proliferated into the peripheral microporous fluorocarbon and provided anchorage between the cornea and prosthesis. Seventeen patients (70.8%) had visual acuity improvements. Mean corrected final visual acuity was 20/100 (range, 20/30 to 20/400). Five anatomic failures occurred in the first 6 months (three extrusions, one dislocation of the optic, and one endophthalmitis). We had one case (4.1%) of treatable glaucoma. We successfully removed four of five retroprosthetic membranes that had occurred. No retinal detachment occurred. The biocompatible inert microporous polymer did not eliminate all mechanical complications associated with a keratoprosthesis.


Trabecular micro-bypass shunt (iStent®): basic science, clinical, and future).
The trabecular bypass stent offers an alternative to filtration surgery. Patients who may be ideal candidates for considering this procedure are those with prior conjunctival surgery; for example, those who had a 360° peritomy from a scleral buckle might not do well with a trabeculectomy and there is no space for a tube. Highly myopic patients do not tolerate hypotony well, and the iSTB may be an option for some of these patients. I have used the iSTB in patients on anticoagulants who could not stop them, and they needed something beyond medications and laser to lower the IOP in subjects with open-angle glaucoma. Young patients, especially those with one eye, who need rapid visual recovery (for instance to return to work) may be good candidates to consider the iSTB as well. Because of the position used for clear corneal cataract surgery, the temporal approach is best for doing these. Therefore, if you are doing cataract surgery on someone who needs a lower IOP, you already are in the correct position to implant the devices. Patients may need some medications after the procedure to lower the IOP to the level desired. The results from Armenia are encouraging, given an IOP of 11.8 mmHg after 2 iSTB stents and taking daily travoprost. These results are difficult to reach even with a trabeculectomy. When selecting your fist patients, avoid those with the congested episcleral veins, look for patients with wide open angles, and if you can see aqueous veins at the slit-lamp it may indicate a viable outflow system. Probably avoid patients with IOPs over 35 mmHg. The micro-invasive trabecular bypass stents offer an alternative surgical intervention for select patients with open-angle glaucoma. Recent studies show that combining these micro-stents with medications can lead to as low of an intraocular pressure (IOP) as is achieved by many more invasive incisional surgeries. The technique is quite precise and learning the procedure is similar to clear corneal phacoemulsification followed by a goniotomy. Long-term data are starting to come in and the safety is favorable. The IOP success appears to be based on the patency of the outflow system for a given patient. Key factors in determining the success involve the placement of trabecular bypass devices into the canal of Schlemm and require a down-stream patency of the collector channel system and a low episcleral venous pressure. Because accessing the collector system may require placement by a patent channel, the placement of two stents, a longer stent with scaffolding or somehow imaging the outflow system may lead to the best control of the IOP.


Tool-use and instrumental learning in the Eurasian jay (Garrulus glandarius).
Recent research with Rooks has demonstrated impressive tool-using abilities in captivity despite this species' classification as a non-tool-user in the wild. Here, we explored whether another non-tool-using corvid, the Eurasian Jay, would be capable of similar feats and investigated the relative contributions of causal knowledge and instrumental conditioning to the birds' performance on the tasks. Five jays were tested on a variety of tasks involving water displacement. Two birds reliably interacted with the apparatuses. In these tasks, both birds showed a preference for inserting stones into a tube containing liquid over a tube containing a solid or a baited 'empty' tube and also for inserting sinkable items over non-sinkable items into a tube of water. To investigate the contribution of instrumental conditioning, subjects were then tested on a series of tasks in which different cues were made available. It was found that, in the absence of any apparent causal cues, these birds showed a clear preference for the rewarded tube when the food incrementally approached with every stone insertion, but not when it simply "appeared" after the correct number of stone insertions. However, it was found that subjects did not prefer to insert stones into a tube rewarded by the incremental approach of food if the available causal cues violated the expectations created by existing causal knowledge (i.e. were counter-intuitive). An analysis of the proportion of correct and incorrect stone insertions made in each trial across tasks offering different types of information revealed that subjects were substantially more successful in experiments in which causal cues were available, but that rate of learning was comparable in all experiments. We suggest that these results indicate that Eurasian jays use the incremental approach of the food reward as a conditioned reinforcer allowing them to solve tasks involving raising the water level and that this learning is facilitated by the presence of causal cues.


Association between Pulse Pressure and Onset of Dementia in an Elderly Korean Population: A Cohort Study.
Objective: There is paucity of studies on the association between pulse pressure and the development of dementia, although this association has already been established. This study aimed at investigating the association between pulse pressure and the onset of dementia. Methods: We used the South Korean National Health Insurance Service claims cohort data to select 149,663 patients without dementia aged ≥60 years. We calculated adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) for dementia using Cox proportional hazard models according to a pulse pressure classification (<50, 50-59, 60-69, 70-79, 80-89, or 90+). Results: Compared to women with pulse pressure <50, those with pulse pressures of 50-59, 60-69, and 90+ had higher HRs for dementia (1.14, 1.22, and 1.03, respectively). These associations were particularly strong in those on Medicaid insurance and from rural regions. However, there were no statistically significant results among men. Conclusions: A higher pulse pressure was associated with an elevated risk of dementia in women aged >60 years, particularly those on Medicaid and from rural regions, possibly due to their inability to access hypertension and other medical treatment. The establishment of dementia indicators will help to guide future health policies for the prevention of dementia.


New observations on meniscal cysts.
The purpose of this study was to determine the incidence of meniscal cysts, assess the frequency of various magnetic resonance (MR) imaging characteristics, and emphasize radiographic observations not commonly reported. A total of 2,095 consecutive knee MR imaging reports from a 22 month period were retrospectively reviewed for the presence of meniscal cysts. Two musculoskeletal radiologists reviewed the cases where cysts were reported. A meniscal cyst was considered present if abnormally increased signal was identified within an enlarged meniscus (i.e., intrameniscal cyst) or if a loculated fluid-intensity lesion with a clear connection to the adjacent meniscus was identified (i.e., parameniscal cyst). Presence or absence of a meniscal tear, intrameniscal and parameniscal signal intensity, patient age, sex, location of meniscal cyst, presence of discoid meniscus, and size of the parameniscal cyst component were recorded. All knee imaging examinations were performed on a 1.5T MR unit. Imaging findings were correlated with arthroscopic reports when available. A total of 167 cases (8.0%) of meniscal cysts were diagnosed in 161 patients. Of the 167 cysts, 69 (41.3%) were located in the lateral meniscus and 98 (58.7%) in the medial meniscus. In 6 patients (3.7%), meniscal cysts were present in both menisci of the same knee. Twelve (7.2%) meniscal cysts were associated with discoid menisci. Ninety-seven (57.8%) meniscal cysts were associated with meniscal tears. Of the total number of meniscal cysts, 104 (62.3%) had a parameniscal cyst. An isolated intrameniscal cyst was present in 63 (37.7%) cases. One hundred (96%) of the parameniscal cyst components were isointense to fluid on T2-weighted FSE images. All cysts exhibited abnormal intrameniscal signal. Only 14 (8.4%) of the intrameniscal components were isointense to fluid on T2-weighted FSE images. The arthroscopic reports of 88 of the 161 (54.7%) patients were available for review and correlation. A tear extending to the articular surface of the meniscus was reported to be present in 74 of the 88 (84%) arthroscopic examinations. Knowledge of the spectrum of findings and the relative frequency of various MR imaging characteristics as well as common potential pitfalls is important to the accurate diagnosis and management of mensical cysts. In particular, controversy exists as to the incidence of articular surface tears in association with meniscal cysts, with some authors reporting greater than 98% correlation with tears. Only 58% of cysts were associated with an articular surface tear. Ninety six percent of the parameniscal signal was isointense to fluid, only 8% of the intramensical signal was isointense to fluid.


Improvement of psychiatrists' clinical knowledge of the treatment guidelines for schizophrenia and major depressive disorders using the 'Effectiveness of Guidelines for Dissemination and Education in Psychiatric Treatment (EGUIDE)' project: A nationwide dissemination, education, and evaluation study.
Although treatment guidelines for pharmacological therapy for schizophrenia and major depressive disorder have been issued by the Japanese Societies of Neuropsychopharmacology and Mood Disorders, these guidelines have not been well applied by psychiatrists throughout the nation. To address this issue, we developed the 'Effectiveness of Guidelines for Dissemination and Education in Psychiatric Treatment (EGUIDE)' integrated education programs for psychiatrists to disseminate the clinical guidelines. Additionally, we conducted a systematic efficacy evaluation of the programs. Four hundred thirteen out of 461 psychiatrists attended two 1-day educational programs based on the treatment guidelines for schizophrenia and major depressive disorder from October 2016 to March 2018. We measured the participants' clinical knowledge of the treatment guidelines using self-completed questionnaires administered before and after the program to assess the effectiveness of the programs for improving knowledge. We also examined the relation between the participants' demographics and their clinical knowledge scores. The clinical knowledge scores for both guidelines were significantly improved after the program. There was no correlation between clinical knowledge and participant demographics for the program on schizophrenia; however, a weak positive correlation was found between clinical knowledge and the years of professional experience for the program on major depressive disorder. Our results provide evidence that educational programs on the clinical practices recommended in guidelines for schizophrenia and major depressive disorder might effectively improve participants' clinical knowledge of the guidelines. These data are encouraging to facilitate the standardization of clinical practices for psychiatric disorders.


Effect of interfacial tension on propagating polymerization fronts.
This paper is devoted to the investigation of polymerization fronts converting a liquid monomer into a liquid polymer. We assume that the monomer and the polymer are immiscible and study the influence of the interfacial tension on the front stability. The mathematical model consists of the reaction-diffusion equations coupled with the Navier-Stokes equations through the convection terms. The jump conditions at the interface take into account the interfacial tension. Simple physical arguments show that the same temperature distribution could not lead to Marangoni instability for a nonreacting system. We fulfill a linear stability analysis and show that interaction of the chemical reaction and of the interfacial tension can lead to an instability that has another mechanism: the heat produced by the reaction decreases the interfacial tension and initiates the liquid motion. It brings more monomer to the reaction zone and increases even more the heat production. This feedback mechanism can lead to the instability if the frontal Marangoni number exceeds a critical value. (c) 2000 American Institute of Physics.


Early versus late onset of therapy. Experiences from animal studies.
Established primary hypertension is characterized by an "upward structural resetting," that encompasses all pressure-exposed cardiovascular sections. This also affects several of the dominating barostat mechanisms, such as those based on the cardiovascular baroreceptors and volume receptors and on the renal excretory function. One major long-term goal in antihypertensive treatment, therefore, is to achieve reversal and, if possible, normalization of this structural resetting, the more so because such reversal would also interrupt the positive feedback interaction with functional excitatory mechanisms. It is, however, very difficult to analyze precisely the rate and extent of structural cardiovascular reversal in humans, and also whether some antihypertensive drugs are in these respects more efficient than others. In this respect, model therapy studies in rat primary hypertension offer several advantages, thanks to the greatly compressed time scale, uniform material, and easily controlled conditions. This facilitates the elucidation of such important problems as the relative merits of early versus late treatment and optimal duration and intensity of treatment, as well as which drug actions are most efficient in normalizing cardiovascular design. The relatively few rat model studies of structural cardiovascular reversal so far performed are outlined briefly. It is concluded that extended studies along these lines would be of great value in clarifying some general principles of antihypertensive treatment that are also likely to be relevant to primary hypertension in humans.


Autologous stem cell transplant in adult multiple sclerosis patients: A study from North India.
Autologous Stem Cell Transplant (ASCT) provides long periods of progression-free-survival in multiple sclerosis (MS). This is an observational study to demonstrate the safety of ASCT in MS patients at a transplant center in North India using a lymphoablative regimen. MS patients > 18 years referred by a neurologist or who came of their own volition were evaluated. Kurtzke Expanded Disability Status Scale (EDSS) score was calculated and those with a score of >7 were excluded. Informed written consent was taken. Mobilization was done with G-CSF with prednisolone to prevent disease flare-up. A minimum of 2 × 106 CD34 cells/kg was collected. Conditioning regimen consisted of rabbit ATG and cyclophosphamide. Rituximab 375 mg/m2 was given to prevent EBV reactivation and disease relapse. Antibiotic prophylaxis was given with levofloxacin, fluconazole, and valacyclovir. Any persistent change in EDSS scores ≥0.5 was considered significant. Twenty patients were included. Seven patients had positive urine cultures prior to transplant and were treated before starting any chemotherapy. Majority patients were women (13/20). All patients developed febrile neutropenia, which was managed as per department policy. There was no mortality. Subjective symptoms improved in all patients. EDSS score improved in 6/19 patients (5/6 with RRMS) with no disease progression in any patient at a median follow-up duration of 242 days. ASCT can be done safely for patients with relatively high EDSS scores with additional precautions for screening for infections. RRMS patients with the active disease show most improvement. SPMS patients may not show significant improvement in the short term.


Mustard-inspired delivery shuttle for enhanced blood-brain barrier penetration and effective drug delivery in glioma therapy.
Effective penetration through the blood-brain barrier (BBB) remains a challenge for the treatment of many brain diseases. In this study, a small molecule, sinapic acid (SA), extracted from mustard, was selected as a novel bioinspired BBB-permeable ligand for efficient drug delivery in glioma treatment. SA was conjugated on the surface of zwitterionic polymer poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC)-encapsulated bovine serum albumin (BSA)-based nanoparticles, yielding nBSA-SA. The PMPC shell serves as a protective layer to prolong the in vivo blood circulation time with a better chance to cross the BBB. Furthermore, temozolomide (TMZ), which can be loaded onto the nanoparticles via electrostatic interactions with acrylic acid (AA) to generate AA-nBSA-SA-TMZ, was applied as an excellent chemotherapeutic drug for glioma therapy. The obtained nanoparticles with a distinct size show great BBB permeability. Through the mechanism study, it was found that the cell internalization of the SA-conjugated nanoparticles is an energy-dependent process with only transient disruption of the BBB. The biological evaluation results unambiguously suggest that drug-loaded nanoparticles can lead to strong apoptosis on the tumor site and increase the median survival time of glioma-bearing mice. Overall, this novel BBB-permeable ligand SA paves the way for the delivery of cargo into the brain and provides a powerful nanoplatform for glioma therapy via intravenous administration.


Comparison of plateletpheresis on three continuous flow cell separators.
Platelet concentrate (PC) remains one of the most important support measures in thrombocytopenic patients. An efficient cell separator is a prerequisite for an optimally functioning apheresis setup. Donor blood count may undergo a temporary reduction after the procedure. The aim was to find the extent of reduction in donor blood count (hemoglobin, hematocrit, white blood cell, and platelet) after plateletpheresis and to evaluate the cell separator for collection efficiency, processing time, and leukoreduction. Two hundred and thirty seven procedures performed on the Amicus (N = 121), Fenwal CS-3000 Plus (N = 50) and Cobe spectra (N = 66) in a one year period were evaluated. The procedures performed on the continuous flow centrifugation (CFC) cell separators and donor blood counts (pre and post donation) done were included in the study. The percent reduction in hemoglobin (HB), hematocrit (HCT), white blood cell (WBC) and platelet count ((PLT ct) was 2.9, 3.1, 9, 30.7 (Mean, N = 237) respectively after the procedure. The post donation PLT ct reduced to < 100x109/L (range 80-100) in five donors (N = 5/237, Amicus). The pre donation PLT ct in them was 150-200x109/L. Collection efficiency (percent) of Amicus (79.3) was better as compared to the other two machines (CS: 62.5, Cobe: 57.5). PC collected on Cobe spectra had <1x106 WBC. The donor pre donation PLT levels had a positive correlation to the product PLT yield (r = 0.30, P = 0.000). Monitoring donor blood counts helps to avoid pheresis induced adverse events. A cautious approach is necessary in donors whose pre donation PLT ct is 150-200x109/L. The main variable in PLT yield is donor PLT ct (pre donation). High collection efficiency is a direct measure of an optimally functioning cell separator.


Sex differences in the brain's dopamine signature of cigarette smoking.
Cigarette smoking is a major public health danger. Women and men smoke for different reasons and cessation treatments, such as the nicotine patch, are preferentially beneficial to men. The biological substrates of these sex differences are unknown. Earlier PET studies reported conflicting findings but were each hampered by experimental and/or analytical limitations. Our new image analysis technique, lp-ntPET (Normandin et al., 2012; Morris et al., 2013; Kim et al., 2014), has been optimized for capturing brief (lasting only minutes) and highly localized dopaminergic events in dynamic PET data. We coupled our analysis technique with high-resolution brain scanning and high-frequency motion correction to create the optimal experiment for capturing and characterizing the effects of smoking on the mesolimbic dopamine system in humans. Our main finding is that male smokers smoking in the PET scanner activate dopamine in the right ventral striatum during smoking but female smokers do not. This finding-men activating more ventrally than women-is consistent with the established notion that men smoke for the reinforcing drug effect of cigarettes whereas women smoke for other reasons, such as mood regulation and cue reactivity. lp-ntPET analysis produces a novel multidimensional endpoint: voxel-level temporal patterns of neurotransmitter release ("DA movies") in individual subjects. By examining these endpoints quantitatively, we demonstrate that the timing of dopaminergic responses to cigarette smoking differs between men and women. Men respond consistently and rapidly in the ventral striatum whereas women respond faster in a discrete subregion of the dorsal putamen.


Varicella in children with cancer: Seventy-seven cases.
The purpose of this study was to characterize varicella in childhood cancer patients. Seventeen of the 77 patients reviewed were in remission and off all therapy for 3 to 22 months. No one in this group died from varicella or had evidence of visceral dissemination. Among the remaining 60 patients, all of whom were receiving anticancer theapy when they developed varicella, 19 (32%) had visceral dissemination and 4 died, for a mortality rate of 7%. Each of the deaths was associated with primary varicella pneumonitis, with or without acute encephalitis. Visceral dissemination was not related to type or status of malignancy or to duration of specific anticancer therapy. Varicella was more likely to disseminate in children with absolute lymphopenia, less than 500 cells per cubic millimeter, than in patients with higher lymphocyte counts. Cessation of anticancer theapy prior to the onset of lesions appeared to lessen the risk of dissemination. These results show that varicella is more severe in cancer patients on therapy than the general population or in patients who have completed therapy, but is not highly fatal.


Replication-dependent sister chromatid recombination in rad1 mutants of Saccharomyces cerevisiae.
Homolog recombination and unequal sister chromatid recombination were monitored in rad1-1/rad1-1 diploid yeast cells deficient for excision repair, and in control cells, RAD1/rad1-1, after exposure to UV irradiation. In a rad1-1/rad1-1 diploid, UV irradiation stimulated much more sister chromatid recombination relative to homolog recombination when cells were irradiated in the G1 or the G2 phases of the cell cycle than was observed in RAD1/rad1-1 cells. Since sister chromatids are not present during G1, this result suggested that unexcised lesions can stimulate sister chromatid recombination events during or subsequent to DNA replication. The results of mating rescue experiments suggest that unexcised UV dimers do not stimulate sister chromatid recombination during the G2 phase, but only when they are present during DNA replication. We propose that there are two types of sister chromatid recombination in yeast. In the first type, unexcised UV dimers and other bulky lesions induce sister chromatid recombination during DNA replication as a mechanism to bypass lesions obstructing the passage of DNA polymerase, and this type is analogous to the type of sister chromatid exchange commonly observed cytologically in mammalian cells. In the second type, strand scissions created by X-irradiation or the excision of damaged bases create recombinogenic sites that result in sister chromatid recombination directly in G2. Further support for the existence of two types of sister chromatid recombination is the fact that events induced in rad1-1/rad1-1 were due almost entirely to gene conversion, whereas those in RAD1/rad1-1 cells were due to a mixture of gene conversion and reciprocal recombination.


VEDA: a web-based virtual environment for dynamic atomic force microscopy.
We describe here the theory and applications of virtual environment dynamic atomic force microscopy (VEDA), a suite of state-of-the-art simulation tools deployed on nanoHUB (www.nanohub.org) for the accurate simulation of tip motion in dynamic atomic force microscopy (dAFM) over organic and inorganic samples. VEDA takes advantage of nanoHUB's cyberinfrastructure to run high-fidelity dAFM tip dynamics computations on local clusters and the teragrid. Consequently, these tools are freely accessible and the dAFM simulations are run using standard web-based browsers without requiring additional software. A wide range of issues in dAFM ranging from optimal probe choice, probe stability, and tip-sample interaction forces, power dissipation, to material property extraction and scanning dynamics over hetereogeneous samples can be addressed.


Prevalence, Disparities, And Determinants Of Primary Cesarean Births Among First-Time Mothers In Mexico.
Mexico has the second-highest prevalence of cesarean deliveries in the Americas, behind Brazil. Having had a previous cesarean delivery is highly predictive of having subsequent cesarean deliveries, yet evidence on the drivers of primary (that is, first-time) cesarean deliveries is sparse. Using 2014 Mexican birth certificate data and performing population-level analyses of data on 600,124 first-time mothers giving birth after at least thirty-seven weeks of gestation, we examined the prevalence and determinants of primary cesarean deliveries. We found a very high prevalence of cesarean deliveries among these women-48.7 percent-and wide variations across insurance coverage types. Enrollees in Seguro Popular, the public health insurance program introduced in 2003 for the previously uninsured and gradually rolled out nationally, had a cesarean rate of 40 percent, while women insured through the Social Security Institute for Civil Servants had a rate of 78 percent. The lower risk of primary cesarean deliveries among Seguro Popular enrollees persisted after adjustment for covariates. Rates of primary cesarean deliveries were particularly high in private birthing facilities for all first-time mothers. Reducing the rate of cesarean deliveries in Mexico will require interventions across types of insurance and birthing facilities and will also require targeted public health messaging.


Chagas disease (Trypanosoma cruzi) and HIV co-infection in Colombia.
Chagas disease is a complex zoonotic pathology caused by the kinetoplastid Trypanosoma cruzi. This parasite presents remarkable genetic variability and has been grouped into six discrete typing units (DTUs). The association between the DTUs and clinical outcome remains unknown. Chagas disease and co-infection with HIV/AIDS has been reported widely in Brazil and Argentina. Herein, we present the molecular analyses from a Chagas disease patient with HIV/AIDS co-infection in Colombia who presented severe cardiomyopathy, pleural effusion, and central nervous system involvement. A mixed infection by T. cruzi genotypes was detected. We suggest including T. cruzi in the list of opportunistic pathogens for the management of HIV patients in Colombia. The epidemiological implications of this finding are discussed.


Lack of effect of gastric acid-reducing agents on the pharmacokinetics of lopinavir/ritonavir in HIV-infected patients.
Recent studies have shown that coadministration of certain protease inhibitors (PIs) with gastric acid-reducing agents results in decreased plasma concentrations of the PI. To assess the effect of acid-reducing agents on lopinavir/ritonavir, data from two clinical trials (n = 38 and 190) were pooled. Both trials randomized antiretroviral-naïve, HIV-infected patients to receive lopinavir/ritonavir 400/100 mg twice-daily or 800/200 mg once-daily in combination with stavudine and lamivudine, or tenofovir and emtricitabine. Concurrent administration of gastric acid-reducing agents including antacids of various brand names, proton pump inhibitors (omeprazole, esomeprazole, lansoprazole, pantoprazole, and rabeprazole), and H(2)-receptor antagonists (ranitidine, famotidine, cimetidine, and nizatidine) was reported in both trials. Lopinavir and ritonavir pharmacokinetic parameters were evaluated. Thirty subjects were considered users of acid-reducing agents at the times of pharmacokinetic evaluation. HIV-infected patients who received gastric acid-reducing agents during administration of lopinavir/ritonavir-based treatment regimens did not appear to have a reduction in lopinavir or ritonavir exposures.


Sexual behaviour in Britain: reported sexually transmitted infections and prevalent genital Chlamydia trachomatis infection.
Studies of the epidemiology of sexually transmitted infections (STI) are largely based on surveillance data. As part of a national survey of sexual attitudes and lifestyles (Natsal 2000) in Britain, we estimated the frequency of self-reported STIs, and the prevalence of urinary Chlamydia trachomatis infection. We did a stratified probability sample survey of 11161 men and women aged 16-44 years in Britain. Computer assisted self-interviews contained items on the nature and timing of previously diagnosed STIs. Half of all sexually experienced respondents aged 18-44 years were invited to provide a urine sample for ligase chain reaction testing for C trachomatis infection. 10.8% of men and 12.6% of women reported ever having an STI, 3.6% of men and 4.1% of women reported ever being diagnosed with genital warts, and 1.4% of men and 3.1% of women reported previous infection with C trachomatis. 76% of men and 57% of women ever diagnosed with an STI had been to a GUM clinic. C trachomatis was found in 2.2% (95% CI 1.5-3.2) of men and 1.5% (95% CI 1.11-2.14) of women with age-specific prevalence being highest among men aged 25-34 (3.1%) and women aged 16-24 years (3.0%). Non-married status, age, and reporting partner concurrency or two or more sexual partners in the past year were independently associated with infection with C trachomatis. We show substantial heterogeneity in distribution of reported STIs, and the demographic and behavioural determinants of prevalent genital chlamydial infection. The results have potentially wide application for proposed chlamydia screening programmes which, given the demonstrated prevalence, must now proactively seek to involve men.


Lupus risk variants in the PXK locus alter B-cell receptor internalization.
Genome wide association studies have identified variants in PXK that confer risk for humoral autoimmune diseases, including systemic lupus erythematosus (SLE or lupus), rheumatoid arthritis and more recently systemic sclerosis. While PXK is involved in trafficking of epidermal growth factor Receptor (EGFR) in COS-7 cells, mechanisms linking PXK to lupus pathophysiology have remained undefined. In an effort to uncover the mechanism at this locus that increases lupus-risk, we undertook a fine-mapping analysis in a large multi-ancestral study of lupus patients and controls. We define a large (257kb) common haplotype marking a single causal variant that confers lupus risk detected only in European ancestral populations and spans the promoter through the 3' UTR of PXK. The strongest association was found at rs6445972 with P < 4.62 × 10(-10), OR 0.81 (0.75-0.86). Using stepwise logistic regression analysis, we demonstrate that one signal drives the genetic association in the region. Bayesian analysis confirms our results, identifying a 95% credible set consisting of 172 variants spanning 202 kb. Functionally, we found that PXK operates on the B-cell antigen receptor (BCR); we confirmed that PXK influenced the rate of BCR internalization. Furthermore, we demonstrate that individuals carrying the risk haplotype exhibited a decreased rate of BCR internalization, a process known to impact B cell survival and cell fate. Taken together, these data define a new candidate mechanism for the genetic association of variants around PXK with lupus risk and highlight the regulation of intracellular trafficking as a genetically regulated pathway mediating human autoimmunity.


Tick community composition in Midwestern US habitats in relation to sampling method and environmental conditions.
The ranges of many tick species are changing due to climate change and human alteration of the landscape. Understanding tick responses to environmental conditions and how sampling method influences measurement of tick communities will improve our assessment of human disease risk. We compared tick sampling by three collection methods (dragging, CO2 trapping and rodent surveys) in adjacent forested and grassland habitats in the lower Midwest, USA, and analyzed the relationship between tick abundance and microclimate conditions. The study areas were within the overlapping ranges of three tick species, which may provide conditions for pathogen exchange and spread into new vectors. Dermacentor variabilis (American dog tick) was found using all methods, Amblyomma americanum (lonestar tick) was found by dragging and CO2 trapping and Ixodes scapularis (blacklegged deer tick) was found only on rodents. Proportion of each species differed significantly among sampling methods. More ticks were found in forests compared to open habitats. Further, more ticks were collected by dragging and from rodents in hotter, drier conditions. Our results demonstrate that multiple sampling methodologies better measure the tick community and that microclimate conditions strongly influence the abundance and activity of individual tick species.


Which strings attached: ethical considerations for selecting appropriate conditionalities in conditional cash transfer programmes.
Conditional cash transfers (CCTs) present a promising approach to simultaneously tackle chronic poverty and poor health. While these programmes clearly embody beneficent aims, questions remain regarding the ethical design of CCTs. Limited guidance exists for the ethical evaluation of the defining feature of these programmes: the conditionalities. Drawing upon prominent public health ethics frameworks and social justice theories, this paper outlines five categories of morally relevant considerations that CCT programme designers should consider when assessing which behaviours or outcomes they select as conditionalities for payment: (1) likelihood of yielding desired health outcomes, (2) risks and burdens, (3) receptivity, (4) attainability and (5) indirect impacts and externalities. When evaluating potential conditionalities across these five categories of considerations, it is important to recognise that not all beneficiaries or subgroups of beneficiaries will fare equally on each. Given that most CCTs aim to reduce inequities and promote long-term health and prosperity for the most disadvantaged, it is critical to apply these considerations with due attention to how different segments of the beneficiary population will be differentially affected. Taken on balance, with due reflection on distributional effects, these five categories represent a comprehensive set of considerations for the moral analysis of specific conditionalities and will help ensure that CCT designers structure programmes in a way that is both morally sound and effective in achieving their goals.


Developing drugs for use before, during and soon after percutaneous coronary intervention.
Percutaneous coronary intervention (PCI) is a milestone for treating coronary artery disease (CAD). Antithrombotic therapy is essential to prevent ischemic complications, including the microvascular no-reflow, while minimizing bleeding events. This overview discusses available and developing drugs for PCI including anticoagulants, antiplatelets and treatment of no-reflow. For years unfractionated heparin (UFH) has been the unique anticoagulant to be used before and during PCI. Enoxaparin showed similar efficacy and safety, yet, based on recent trials, bivalirudin has been shown to have some benefits, particularly for patients with ST-segment elevation myocardial infarction (STEMI). The evidence concerning new anticoagulants is still preliminary, except for new oral anticoagulants, particularly rivaroxaban that showed intriguing findings and is currently under investigation. Dual antiplatelet therapy (DAPT) is the standard of care after PCI, but new developments have recently emerged. Indeed, ticagrelor and prasugrel are currently recommended over clopidogrel due to their significant reduction of ischemic events in acute coronary syndrome (ACS) whereas clopidogrel remains the choice in stable CAD. Among new agents, vorapaxar and cangrelor showed positive but limited evidence and might be considered at least in selected patients. Conversely, evidence on effective treatments for no-reflow remains limited and would require future dedicated research.


Cell-free assembly of rough and smooth endoplasmic reticulum.
Smooth endoplasmic reticulum assembly was studied in a cell-free system using thin-section and freeze-fracture electron microscopy. Incubation of rat hepatocyte rough and smooth microsomes in the presence of ATP, GTP, cytosol (Xenopus egg) and an ATP-regenerating system led to assembly of membrane networks comprising a central core of interconnecting smooth tubules continuous with peripherally located rough membrane cisternae. Glucose-6-phosphatase cytochemistry confirmed the endoplasmic reticulum origin of the reconstituted membranes. When both ATP and GTP were omitted from the incubation medium, or when GTP was replaced by a variety of nucleotide analogues, including GTP gamma S, membrane aggregates contained only unfused microsomes. The presence of GTP alone stimulated assembly of rough membrane cisternae but had no effect on smooth membranes. Smooth tubule formation occurred independent of cytosol and an ATP-regenerating system, but did require GTP and ATP. Omission of ATP, or replacement of this nucleotide with a variety of analogues, including ATP gamma S, prevented tubule formation but did not affect the assembly of the rough membrane cisternae. Morphometric studies revealed sequential formation of rough membrane cisternae (0-60 minutes) followed by appearance of interconnecting smooth tubules (> 60 minutes). The amount of rough membrane cisternae per membrane network diminished with time after 60 minutes; that of smooth tubules increased. Thus GTP is required for reconstitution of rough membrane cisternae, both GTP and ATP are required for smooth tubule formation, and assembly of smooth tubules occurs as an outgrowth (i.e. via tubulation) from rough membranes.


Plasma Soluble CD163 Level Independently Predicts All-Cause Mortality in HIV-1-Infected Individuals.
CD163, a monocyte- and macrophage-specific scavenger receptor, is shed as soluble CD163 (sCD163) during the proinflammatory response. Here, we assessed the association between plasma sCD163 levels and progression to AIDS and all-cause mortality among individuals infected with human immunodeficiency virus type 1 (HIV). Plasma sCD163 levels were measured in 933 HIV-infected individuals. Hazard ratios (HRs) with 95% confidence intervals (CIs) associated with mortality were computed by Cox proportional hazards regression. At baseline, 86% were receiving antiretroviral treatment, 73% had plasma a HIV RNA level of <50 copies/mL, and the median CD4(+) T-cell count was 503 cells/µL. During 10.5 years of follow-up, 167 (17.9%) died. Plasma sCD163 levels were higher in nonsurvivors than in survivors (4.92 mg/L [interquartile range {IQR}, 3.29-8.65 mg/L] vs 3.16 mg/L [IQR, 2.16-4.64 mg/L]; P = .0001). The cumulative incidence of death increased with increasing plasma sCD163 levels, corresponding to a 6% or 35% increased risk of death for each milligram per liter or quartile increase, respectively, in baseline plasma sCD163 level (adjusted HR, 1.06 [95% CI, 1.03-1.09] and 1.35 [95% CI, 1.13-1.63], respectively). Plasma sCD163 was an independent marker of all-cause mortality in a cohort of HIV-infected individuals, suggesting that monocyte/macrophage activation may play a role in HIV pathogenesis and be a target of intervention.


Deciphering the role of endogenous opioids in high-frequency TENS using low and high doses of naloxone.
Previous human studies have shown that the analgesic effect of high-frequency TENS could not be reversed by low doses of naloxone. The aim of the present study was to reinvestigate the possible contribution of opioid receptors to high-frequency TENS analgesia by using low (0.02 mg/kg) and high (0.14 mg/kg) doses of naloxone. Naloxone (high and low doses) and saline were administered intravenously to young healthy adults using a triple-blind randomized cross-over design. For each visit, TENS (100 Hz, 60 μs) was applied for 25 min to the external surface of the left ankle. TENS intensity was adjusted to obtain strong but comfortable (innocuous) paresthesias. Experimental pain was evoked with a 1 cm(2) thermode applied on the lateral aspect of the left heel. Subjective pain scores were obtained before, during and after TENS. Because preliminary analyses showed that the order of presentation affected the pattern of results, only the first visit of every participant could be analyzed without fear of contamination from possible carry-over effects. These revealed that TENS maintained its analgesic properties following the injection of saline (p<.001) and the injection of a low dose of naloxone (p<.05). However, when a high dose of naloxone was administered, TENS analgesia was completely blocked (p=.20). These results suggest that high-frequency TENS involves opioid receptors. An insufficient amount of opioid antagonist likely prevented previous human studies from discovering the importance of opioid receptors in producing high-frequency TENS analgesia.


Fast Self-Healing of Polyelectrolyte Multilayer Nanocoating and Restoration of Super Oxygen Barrier.
A self-healable gas barrier nanocoating, which is fabricated by alternate deposition of polyethyleneimine (PEI) and polyacrylic acid (PAA) polyelectrolytes, is demonstrated in this study. This multilayer film, with high elastic modulus, high glass transition temperature, and small free volume, has been shown to be a super oxygen gas barrier. An 8-bilayer PEI/PAA multilayer assembly (≈700 nm thick) exhibits an oxygen transmission rate (OTR) undetectable to commercial instrumentation (<0.005 cc (m-2 d-1 atm-1 )). The barrier property of PEI/PAA nanocoating is lost after a moderate amount of stretching due to its rigidity, which is then completely restored after high humidity exposure, therefore achieving a healing efficiency of 100%. The OTR of the multilayer nanocoating remains below the detection limit after ten stretching-healing cycles, which proves this healing process to be highly robust. The high oxygen barrier and self-healing behavior of this polymer multilayer nanocoating makes it ideal for packaging (food, electronics, and pharmaceutical) and gas separation applications.


Subtype-specific sorting of the ETA endothelin receptor by a novel endocytic recycling signal for G protein-coupled receptors.
We have previously reported that endocytic sorting of ET(A) endothelin receptors to the recycling pathway is dependent on a signal residing in the cytoplasmic carboxyl-terminal region. The aim of the present work was to characterize the carboxyl-terminal recycling motif of the ET(A) receptor. Assay of truncation mutants of the ET(A) receptor with increasing deletions of the carboxyl-terminal tail revealed that amino acids 390 to 406 contained information critical for the ability of the receptor to recycle. This peptide sequence displayed significant sequence similarity to several protein segments confirmed by X-ray crystallography to adopt antiparallel beta-strand structures (beta-finger). One of these segments was the beta-finger motif of neuronal nitric-oxide synthase reported to function as an internal PDZ (postsynaptic density-95/disc-large/zona occludens) domain-binding ligand. Based on these findings, the three-dimensional structure of the recycling motif of ET(A) receptor was predicted to attain a beta-finger conformation acting as an internal PDZ ligand. Site-directed mutagenesis at residues that would be crucial to the structural integrity of the putative beta-finger conformation or PDZ ligand function prevented recycling of the ET(A) receptor. Analysis of more than 300 G protein-coupled receptors (GPCRs) identified 35 different human GPCRs with carboxylterminal sequence patterns that fulfilled the structural criteria of an internal PDZ ligand. Among these are several receptors reported to follow a recycling pathway. In conclusion, recycling of ET(A) receptor is mediated by a motif with the structural characteristics of an internal PDZ ligand. This structural motif may represent a more general principle of endocytic sorting of GPCRs.


Bacterial injectisomes: needle length does matter.
Many pathogenic bacteria use a type III secretion nanomachine (an injectisome) to deliver virulence proteins into the cytosol of their eukaryotic host cells. Most injectisomes possess a stiff needlelike structure of a genetically defined length. We found that a minimal needle length was required for efficient functioning of the Yersinia enterocolitica injectisome. This minimal needle length correlated with the length of the major adhesin at the bacterial surface. The needle may be required for triggering type III secretion, and its length may have evolved to match specific structures at the bacterial and host cell surfaces.


Evidence for coevolution of sociality and relative brain size in three orders of mammals.
As the brain is responsible for managing an individual's behavioral response to its environment, we should expect that large relative brain size is an evolutionary response to cognitively challenging behaviors. The "social brain hypothesis" argues that maintaining group cohesion is cognitively demanding as individuals living in groups need to be able to resolve conflicts that impact on their ability to meet resource requirements. If sociality does impose cognitive demands, we expect changes in relative brain size and sociality to be coupled over evolutionary time. In this study, we analyze data on sociality and relative brain size for 206 species of ungulates, carnivores, and primates and provide, for the first time, evidence that changes in sociality and relative brain size are closely correlated over evolutionary time for all three mammalian orders. This suggests a process of coevolution and provides support for the social brain theory. However, differences between taxonomic orders in the stability of the transition between small-brained/nonsocial and large-brained/social imply that, although sociality is cognitively demanding, sociality and relative brain size can become decoupled in some cases. Carnivores seem to have been especially prone to this.


Cellular and molecular bases of skeletal regeneration: what can we learn from genetic mouse models?
Although bone repairs through a very efficient regenerative process in 90% of the patients, many factors can cause delayed or impaired healing. To date, there are no reliable biological parameters to predict or diagnose bone repair defects. Orthopedic surgeons mostly base their diagnoses on radiographic analyses. With the recent progress in our understanding of the bone repair process, new methods may be envisioned. Animal models have allowed us to define the key steps of bone regeneration and the biological and mechanical factors that may influence bone healing in positive or negative ways. Most importantly, small animal models such as mice have provided powerful tools to apprehend the genetic bases of normal and impaired bone healing. The current review presents a state of the art of the genetically modified mouse models that have advanced our understanding of the cellular and molecular components of bone regeneration and repair. The review illustrates the use of these models to define the role of inflammation, skeletal cell lineages, signaling pathways, the extracellular matrix, osteoclasts and angiogenesis. These genetic mouse models promise to change the field of orthopedic surgery to help establish genetic predispositions for delayed repair, develop models of non-union that mimic the human conditions and elaborate new therapeutic approaches to enhance bone regeneration.


Dietary flavonoid and lignan intake and gastric adenocarcinoma risk in the European Prospective Investigation into Cancer and Nutrition (EPIC) study.
Several experimental studies have suggested potential anticarcinogenic effects of flavonoids, although epidemiologic evidence for the impact of dietary flavonoids on risk of gastric cancer (GC) is limited. We investigated the association between intake of dietary flavonoids and lignans and incident GC. The study followed 477,312 subjects (29.8% men) aged 35-70 y from 10 European countries who participated in the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Validated dietary questionnaires and lifestyle information were collected at baseline. A food-composition database on flavonoids and lignans was compiled by using data from USDA and Phenol-Explorer databases. During an average follow-up of 11 y, 683 incident GC cases (57.8% men) were mostly validated by a panel of pathologists and used in this analysis. We observed a significant inverse association between total flavonoid intake and GC risk in women (HR: 0.81; 95% CI: 0.70, 0.94; for the continuous variable after log₂ transformation) but not in men (HR: 0.97; 95% CI: 0.85, 1.09). In women, significant inverse associations with GC risk were also observed for intakes of some flavonoid subgroups (anthocyanidins, flavonols, flavones, and flavanols), particularly with intestinal type tumors for total flavonoid and flavanol intakes (P-heterogeneity < 0.1). After stratification by smoking status and sex, there was no significant heterogeneity in these associations between ever- and never-smokers. Total dietary flavonoid intake is associated with a significant reduction in the risk of GC in women.


Assessment of background radioactivity level for Gaziantep region of southeastern Turkey.
This study presents the measurement results of environmental radioactivity levels for Gaziantep, an industrial and trade centre in the southeastern part of Turkey. The outdoor gamma absorbed dose was measured as 50.1 nGy h(-1), corresponding to a total gamma radiation level (of terrestrial and cosmic origin) of 61.5 microSv y(-1). The activity concentrations in the surface soil samples collected from the study area were determined as 25.2, 23.7 and 289.2 Bq kg(-1) for the natural radionuclides (238)U, (232)Th and (40)K, respectively, and 8.02 Bq kg(-1) for the fission product (137)Cs. These natural radioactivity sources result in a terrestrial gamma level of 46.9 microSv y(-1). The drinking water samples collected from the region carry an average of 0.0493 Bq l(-1) of gross alpha and 0.1284 Bq l(-1) of gross beta activities, which together cause an annual effective dose of 1.83 microSv. The measurement results obtained in this study indicate that the region has a background radiation level that is within the natural limits and shows no significant departure from the other parts of the country.


Emergence of the virulence-associated PB2 E627K substitution in a fatal human case of highly pathogenic avian influenza virus A(H7N7) infection as determined by Illumina ultra-deep sequencing.
Avian influenza viruses are capable of crossing the species barrier and infecting humans. Although evidence of human-to-human transmission of avian influenza viruses to date is limited, evolution of variants toward more-efficient human-to-human transmission could result in a new influenza virus pandemic. In both the avian influenza A(H5N1) and the recently emerging avian influenza A(H7N9) viruses, the polymerase basic 2 protein (PB2) E627K mutation appears to be of key importance for human adaptation. During a large influenza A(H7N7) virus outbreak in the Netherlands in 2003, the A(H7N7) virus isolated from a fatal human case contained the PB2 E627K mutation as well as a hemagglutinin (HA) K416R mutation. In this study, we aimed to investigate whether these mutations occurred in the avian or the human host by Illumina Ultra-Deep sequencing of three previously uninvestigated clinical samples obtained from the fatal case. In addition, we investigated three chicken samples, two of which were obtained from the source farm. Results showed that the PB2 E627K mutation was not present in any of the chicken samples tested. Surprisingly, the avian samples were characterized by the presence of influenza virus defective RNA segments, suggestive for the synthesis of defective interfering viruses during infection in poultry. In the human samples, the PB2 E627K mutation was identified with increasing frequency during infection. Our results strongly suggest that human adaptation marker PB2 E627K has emerged during virus infection of a single human host, emphasizing the importance of reducing human exposure to avian influenza viruses to reduce the likelihood of viral adaptation to humans.


In vivo 1H MRS of human gallbladder bile at 3 T in one and two dimensions: detection and quantification of major biliary lipids.
In vitro (1)H MRS of human bile has shown potential in the diagnosis of various hepatopancreatobiliary (HPB) diseases. Previously, in vivo (1)H MRS of human bile in gallbladder using a 1.5 T scanner demonstrated the possibility of quantification of choline-containing phospholipids (chol-PLs). However, other lipid components such as bile acids play an important role in the pathophysiology of the HPB system. We have employed a higher magnetic field strength (3 T), and a custom-built receive array coil, to improve the quality of in vivo (1)H MRS of human bile in the gallbladder. We obtained significant improvement in the quality of 1D spectra (17 healthy volunteers) using a respiratory-gated PRESS sequence with well distinguished signals for total bile acids (TBAs) plus cholesterol resonating at 0.66 ppm, taurine-conjugated bile acids (TCBAs) at 3.08 ppm, chol-PLs at 3.22 ppm, glycine-conjugated bile acids (GCBAs) at 3.74 ppm, and the amide proton (-NH) arising from GCBAs and TCBAs in the region 7.76-8.05 ppm. The peak areas of these signals were measured by deconvolution, and subsequently the molar concentrations of metabolites were estimated with good accuracy, except for that of TBAs plus cholesterol. The concentration of TBAs plus cholesterol was overestimated in some cases, which could be due to lipid contamination. In addition, we report the first 2D L-COSY spectra of human gallbladder bile in vivo (obtained in 15 healthy volunteers). 2D L-COSY spectra will be helpful in differentiating various biliary chol-PLs in pathological conditions of the HPB system.