AAV9 delivering a modified human Mullerian inhibiting substance as a gene therapy in patient-derived xenografts of ovarian cancer.
To improve ovarian cancer patient survival, effective treatments addressing chemoresistant recurrences are particularly needed. Mullerian inhibiting substance (MIS) has been shown to inhibit the growth of a stem-like population of ovarian cancer cells. We have recently engineered peptide modifications to human MIS [albumin leader Q425R MIS (LRMIS)] that increase production and potency in vitro and in vivo. To test this novel therapeutic peptide, serous malignant ascites from highly resistant recurrent ovarian cancer patients were isolated and amplified to create low-passage primary cell lines. Purified recombinant LRMIS protein successfully inhibited the growth of cancer spheroids in vitro in a panel of primary cell lines in four of six patients tested. Adeno-associated virus (AAV) -delivered gene therapy has undergone a clinical resurgence with a good safety profile and sustained gene expression. Therefore, AAV9 was used as a single i.p. injection to deliver LRMIS to test its efficacy in inhibiting growth of palpable tumors in patient-derived ovarian cancer xenografts from ascites (PDXa). AAV9-LRMIS monotherapy resulted in elevated and sustained blood concentrations of MIS, which significantly inhibited the growth of three of five lethal chemoresistant serous adenocarcinoma PDXa models without signs of measurable or overt toxicity. Finally, we tested the frequency of MIS type II receptor expression in a tissue microarray of serous ovarian tumors by immunohistochemistry and found that 88% of patients bear tumors that express the receptor. Taken together, these preclinical data suggest that AAV9-LRMIS provides a potentially well-tolerated and effective treatment strategy poised for testing in patients with chemoresistant serous ovarian cancer.


The role of type I hypersensitivity reaction and IgE-mediated mast cell activation in acute interstitial nephritis.
Acute interstitial nephritis (AIN) presents with acute kidney injury, with evidence of interstitial inflammation and tubulitis on histology, and the presence of fever, rash, and eosinophiluria. Although the pathogenesis of this disease is not well understood, cell-mediated immunity is thought to play a major role. We hypothesized that IgE mediated mast cell activation is also involved in the pathogenesis of renal injury in AIN. 28 patients, with biopsy proven AIN over a 5-year period, were included in this study. Clinical data, including renal outcomes and the etiologies of AIN, were evaluated in all patients. Available tissues (renal biopsy) from 26 of the patients were stained for β-tryptase (marker for mast cell degranulation), IgE, IL-16, and CD3. A negative control for immunostaining was included. Samples from all 26 individuals stained positive for β-tryptase (mean of 11.16 cells/high power field), IgE (mean average of 0.68 cells/HPF), IL-16 (28% of the interstitium), and CD3 (33% of the interstitium). Acute interstitial nephritis was due to medication in 73%, systemic disease in 15%, and unknown (idiopathic) in 12% of the cases. 86% of patients were treated with corticosteroids. 18% required acute inpatient dialysis, with 7% remaining on dialysis longterm. Our study suggests that IgE and mast cell activation may play a role in the pathogenesis of AIN.


Deep evolutionary lineages in a Western Mediterranean snake (Vipera latastei/monticola group) and high genetic structuring in Southern Iberian populations.
Phylogeographic studies during the last decade confirmed an internal complexity of the Iberian Peninsula and northern Maghreb as refugial areas during the Miocene to Pleistocene period. Species with low vagility that experienced the complex climatic and palaeogeographic processes occurred in the Western Mediterranean Basin are excellent candidates to study the extent of lineage diversification in this region. We applied phylogenetic analyses based on mitochondrial data to infer the evolutionary history of Vipera latastei/monticola and identify the major biogeographic events structuring the genetic diversity within this group. We obtained a well-resolved phylogeny, with four highly divergent lineages (one African and three Iberian) that originated in the Tertiary. Coalescence-based estimations suggest that the differentiation of the four major lineages in V. latastei/monticola corresponds to the Messinian salinity crisis and the reopening of the Strait of Gibraltar during the Miocene. Subsequent Pliocene and Pleistocene climatic oscillations continued to isolate both Iberian and Maghrebian populations and led to a high genetic structuring in this group, particularly in Southern Iberia, a complex palaeogeographic and topographic region with high endemism levels. This study does not support the current taxonomy of the group, thus suggesting that an integrative evaluation of Iberian and African populations is needed to resolve its systematics.


Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin.
This protocol describes a simple and efficient way to label specific cell surface proteins with biophysical probes on mammalian cells. Cell surface proteins tagged with a 15-amino acid peptide are biotinylated by Escherichia coli biotin ligase (BirA), whereas endogenous proteins are not modified. The biotin group then allows sensitive and stable binding by streptavidin conjugates. This protocol describes the optimal use of BirA and streptavidin for site-specific labeling and also how to produce BirA and monovalent streptavidin. Streptavidin is tetravalent and the cross-linking of biotinylated targets disrupts many of streptavidin's applications. Monovalent streptavidin has only a single functional biotin-binding site, but retains the femtomolar affinity, low off-rate and high thermostability of wild-type streptavidin. Site-specific biotinylation and streptavidin staining take only a few minutes, while expression of BirA takes 4 d and expression of monovalent streptavidin takes 8 d.


Starting early: obesity prevention during infancy.
Obesity prevalence among infants and young children has increased rapidly during the past 4 decades, a disturbing trend given early obesity's association with later life obesity and its comorbidities. Fortunately, infancy is a period of great behavioral and metabolic plasticity offering numerous targets for preventive interventions. Modifiable factors that may affect early rapid weight gain and obesity risk include infant sleep duration, feeding to soothe infant distress, and the introduction of solid foods and transitional feeding. We discuss evidence linking these factors to weight outcomes, as well as results from behavioral obesity interventions in infancy, from our laboratory and others'. For example, in a recent pilot intervention, we focused on helping new mothers address three areas of infant behavior hypothesized to affect weight gain and early obesity risk: infant sleeping, crying, and feeding. First-time mothers were randomly assigned to receive either a Soothe/Sleep intervention, an Introduction of Solids intervention, both interventions, or no interventions. The interventions were delivered via home visits and showed positive effects on infant behaviors and weight outcomes at 1 year. Based on evidence from such pilot interventions, we assess the plausibility of targeting behavioral factors in infancy and suggest next steps for early prevention research.


Image reproduction with interactive graphics.
Software application or development in optical image digital data processing requires a fast, good quality, yet inexpensive hard copy of processed images. To achieve this, a Cambo camera with an f 2.8/150-mm Xenotar lens in a Copal shutter having a Graflok back for 4 x 5 Polaroid type 57 pack-film has been interfaced to an existing Adage, AGT-30/Electro-Mechanical, EMR 6050 graphic computer system. Time-lapse photography in conjunction with a log to linear voltage transformation has resulted in an interactive system capable of producing a hard copy in 54 sec (Fig. 1). The interactive aspect of the system lies in a Tektronix 4002 graphic computer terminal and its associated hard copy unit (Fig. 2).


Properties of the statpac visual field index.
Purpose. To compare the properties of the visual field index (VFI) to those of mean deviation (MD) in patients with glaucoma. Methods. MD and VFI were calculated in data obtained from an ongoing longitudinal study in which patients with glaucoma (N = 109, 204 eyes) were observed for 9.8 years (median, 21 tests) with static automated perimetry. MD and VFI were compared in one test of each eye, and a subset of 30 tests were selected to compare the VFI with the judgments of eight experts who judged the percentage of the remaining visual field. In series of tests obtained over time, rates of change, statistical significance, evidence of nonlinearity, and variability were compared between both indices. Results. In single tests, MD and VFI were closely related (r = 0.88, P < 0.001). The relationship between both indices appeared linear, except in visual fields with MDs better than -5.0 dB where 29 (22%) of 129 eyes exhibited a ceiling effect (VFI = 100%). Based on this relationship, the predicted VFIs for visual fields with MDs of -5, -10, and -15 dB were 91%, 76%, and 60%, respectively. The percentage of remaining visual field suggested by the VFI exceeded the range of the experts' subjective judgments in 16 (53%) of 30 eyes. In series of tests obtained over time, rates of change with the two indices were closely related (r = 0.79, P < 0.001), and statistically significant reductions over time (P < 0.05) occurred in a similar number of eyes (92 [45%] with MD, and 87 [43%] with VFI). Of the 105 eyes with statistically significant (P < 0.05) negative trend in either MD or VFI, 74 (70%) showed such trends with both indices (κ = 0.69). The variability of MD and VFI increased with damage, and there was no evidence that change over time was more linear with VFI than with MD. Conclusions. The VFI provides a simple and understandable metric of visual field damage, but its estimates of remaining visual field were more optimistic than those of the experts. Rates of change over time with both indices were closely related, but the reliance of the VFI on pattern deviation probability maps caused a ceiling effect that may have reduced its sensitivity to change in eyes with early damage. In this group of patients there was no evidence to suggest that the VFI is either superior or inferior to the MD as a summary measure of visual field damage.


Comparison of Experiences in Two Birth Cohorts Comprising Young Families with Children under Four Years during the Initial COVID-19 Lockdown in Australia and the UK: A Qualitative Study.
This study aims to understand the experience and impact of the initial COVID-19 lockdown in young families with children aged below 4 years. Free text questions were administered to participants in the ORIGINS (Australia) and Born in Bradford (UK) cohort studies to collect qualitative information on worries, concerns and enjoyable experiences during the pandemic. A total of 903 (400 for ORIGINS and 503 for BiB) participants completed the two surveys during April 2020. Despite varying in geography, levels of socio-economic disadvantage and their situational context during the pandemic, respondents from both cohorts reported similar worries and challenges during the lockdown period, including: employment/finances, health anxiety, mental health and social isolation, caring for children and child development. Families across the globe experienced both positive and negative immediate impacts of COVID-19. Population-based data can be used to inform the development of support services, public health campaigns and universal interventions to assist families in future health crises.


Neonatal Plasma Transfusion: An Evidence-Based Review.
Several clinical scenarios for plasma transfusion are repeatedly identified in audits, including treatment of bleeding in association with laboratory evidence of coagulopathy, correction of disseminated intravascular coagulation, prevention of intraventricular hemorrhage, management of critically ill neonates (eg, during sepsis or as a volume expander), or correction of markers of prolonged coagulation in the absence of bleeding. The findings of at least one national audit of transfusion practice indicated that almost half of plasma transfusions are given to neonates with abnormal coagulation values with no evidence of active bleeding, despite the limited evidence base to support the effectiveness of this practice. Plasma transfusions to neonates should be considered in the clinical context of bleeding (eg, vitamin K dependent), disseminated intravascular coagulation, and very rare inherited deficiencies of coagulation factors. There seems to be no role for prophylactic plasma to prevent intraventricular hemorrhage or for use as a volume expander.


Phase-field theories for mathematical modeling of biological membranes.
Biological membranes are complex structures whose mechanics are usually described at a mesoscopic level, such as the Helfrich bending theory. In this article, we present the phase-field methods, a useful tool for studying complex membrane problems which can be applied to very different phenomena. We start with an overview of the general theory of elasticity, paying special attention to its derivation from a molecular scale. We then study the particular case of membrane elasticity, explicitly obtaining the Helfrich bending energy. Within the framework of this theory, we derive a phase-field model for biological membranes and explore its physical basis and interpretation in terms of membrane elasticity. We finally explain three examples of applications of these methods to membrane related problems. First, the case of vesicle pearling and tubulation, when lipidic vesicles are exposed to the presence of hydrophobic polymers that anchor to the membrane, inducing a shape instability. Finally, we study the behavior of red blood cells while flowing in narrow microchannels, focusing on the importance of membrane elasticity to the cell flow capabilities.


Impact of point-of-care CD4 testing on linkage to HIV care: a systematic review.
Point-of-care testing for CD4 cell count is considered a promising way of reducing the time to eligibility assessment for antiretroviral therapy (ART) and of increasing retention in care prior to treatment initiation. In this review, we assess the available evidence on the patient and programme impact of point-of-care CD4 testing. We searched nine databases and two conference sites (up until 26 October 2013) for studies reporting patient and programme outcomes following the introduction of point-of-care CD4 testing. Where appropriate, results were pooled using random-effects methods. Fifteen studies, mainly from sub-Saharan Africa, were included for review, providing evidence for adults, adolescents, children and pregnant women. Compared to conventional laboratory-based testing, point-of-care CD4 testing increased the likelihood of having CD4 measured [odds ratio (OR) 4.1, 95% CI 3.5-4.9, n=2] and receiving a CD4 result (OR 2.8, 95% CI 1.5-5.6, n=6). Time to being tested was significantly reduced, by a median of nine days; time from CD4 testing to receiving the result was reduced by as much as 17 days. Evidence for increased treatment initiation was mixed. The results of this review suggest that point-of-care CD4 testing can increase retention in care prior to starting treatment and can also reduce time to eligibility assessment, which may result in more eligible patients being initiated on ART.


A study of the workforce in emergency medicine: 2007.
This study was undertaken to describe the current status of the emergency medicine workforce in the United States. Surveys were distributed in 2008 to 2619 emergency department (ED) medical directors and nurse managers in hospitals in the 2006 American Hospital Association database. Among ED medical directors, 713 responded, for a 27.2% response rate. Currently, 65% of practicing emergency physicians are board certified by the American Board of Emergency Medicine or the American Osteopathic Board of Emergency Medicine. Among those leaving the practice, the most common reasons cited for departure include geographic relocation (46%) and better pay (29%). Approximately 12% of the ED physician workforce is expected to retire in the next 5 years. Among nurse managers, 548 responded, for a 21% response rate. Many nurses (46%) have an associate degree as their highest level of education, 28% have a BSN, and 3% have a graduate degree (MSN or higher). Geographic relocation (44%) is the leading reason for changing employment. Emergency department annual volumes have increased by 49% since 1997, with a mean ED volume of 32 281 in 2007. The average reported ED length of stay is 158 minutes from registration to discharge and 208 minutes from registration to admission. Emergency department spent an average of 49 hours per month in ambulance diversion in 2007. Boarding is common practice, with an average of 318 hours of patient boarding per month. In the past 10 years, the number of practicing emergency physicians has grown to more than 42 000. The number of board-certified emergency physicians has increased. The number of annual ED visits has risen significantly.


Isochromosome 17q in blast crisis of chronic myeloid leukemia and in other hematologic malignancies is the result of clustered breakpoints in 17p11 and is not associated with coding TP53 mutations.
An isochromosome of the long arm of chromosome 17, i(17q), is the most frequent genetic abnormality observed during the disease progression of Philadelphia chromosome-positive chronic myeloid leukemia (CML), and has been described as the sole anomaly in various other hematologic malignancies. The i(17q) hence plays a presumably important pathogenetic role both in leukemia development and progression. This notwithstanding, the molecular consequences of this abnormality have not been investigated in detail. We have analyzed 21 hematologic malignancies (8 CML in blast crisis, 8 myelodysplastic syndromes [MDS], 2 acute myeloid leukemias, 2 chronic lymphocytic leukemias, and 1 acute lymphoblastic leukemia) with i(17q) by fluorescence in situ hybridization (FISH). Using a yeast artificial chromosome (YAC) contig, derived from the short arm of chromosome 17, all cases were shown to have a breakpoint in 17p. In 12 cases, the breaks occurred within the Smith-Magenis Syndrome (SMS) common deletion region in 17p11, a gene-rich region which is genetically unstable. In 10 of these 12 cases, we were able to further map the breakpoints to specific markers localized within a single YAC clone. Six other cases showed breakpoints located proximally to the SMS common deletion region, but still within 17p11, and yet another case had a breakpoint distal to this region. Furthermore, using chromosome 17 centromere-specific probes, it could be shown that the majority of the i(17q) chromosomes (11 of 15 investigated cases) were dicentric, ie, they contained two centromeres, strongly suggesting that i(17q) is formed through an intrachromosomal recombination event, and also implicating that the i(17q), in a formal sense, should be designated idic(17)(p11). Because i(17q) formation results in loss of 17p material, potentially uncovering the effect of a tumor suppressor on the remaining 17p, the occurrence of TP53 mutations was studied in 17 cases by sequencing the entire coding region. In 16 cases, no TP53 mutations were found, whereas one MDS displayed a homozygous deletion of TP53. Thus, our data suggest that there is no association between i(17q) and coding TP53 mutations, and that another tumor suppressor gene(s), located in proximity of the SMS common deletion region, or in a more distal location, is of pathogenetic importance in i(17q)-associated leukemia.


A Narrative Phenomenological Approach to Transformative Learning: Lessons From Occupational Therapy Reasoning in Educational Practice.
Transformative learning involves the questioning of worldviews and underpins shifts in values and identity that are integral to critical occupational therapy practices. Cognitive theories of transformative learning name, but do not address, the experiential dimensions of transformation. The aim of this article is to conceptualize transformative learning from the perspective of narrative phenomenology in occupational therapy. An argument is presented that draws on research in occupational therapy professional reasoning and that makes visible the dimensions of transformation that are inherently experiential and meaning oriented. Three key concepts for a transformative pedagogy are defined and illustrated: scenes, emplotment, and embodied metaphors. The article concludes with the types of learning objectives for which this approach is suited and the pedagogical values that underpin these teaching practices. This article adds to the domain of health care education by defining and illustrating experiential and meaning-based practices as signature pedagogies for transformative learning.


Expression of calcium release-activated and voltage-gated calcium channels genes in peripheral blood mononuclear cells is altered in pregnancy and in type 1 diabetes.
Calcium (Ca2+) is an important ion in physiology and is found both outside and inside cells. The intracellular concentration of Ca2+ is tightly regulated as it is an intracellular signal molecule and can affect a variety of cellular processes. In immune cells Ca2+ has been shown to regulate e.g. gene transcription, cytokine secretion, proliferation and migration. Ca2+ can enter the cytoplasm either from intracellular stores or from outside the cells when Ca2+ permeable ion channels in the plasma membrane open. The Ca2+ release-activated (CRAC) channel is the most prominent Ca2+ ion channel in the plasma membrane. It is formed by ORAI1-3 and the channel is opened by the endoplasmic reticulum Ca2+ sensor proteins stromal interaction molecules (STIM) 1 and 2. Another group of Ca2+ channels in the plasma membrane are the voltage-gated Ca2+ (CaV) channels. We examined if a change in immunological tolerance is accompanied by altered ORAI, STIM and CaV gene expression in peripheral blood mononuclear cells (PBMCs) in pregnant women and in type 1 diabetic individuals. Our results show that in pregnancy and type 1 diabetes ORAI1-3 are up-regulated whereas STIM1 and 2 are down-regulated in pregnancy but only STIM2 in type 1 diabetes. Expression of L-, P/Q-, R- and T-type voltage-gated Ca2+ channels was detected in the PBMCs where the CaV2.3 gene was up-regulated in pregnancy and type 1 diabetes whereas the CaV 2.1 and CaV3.2 genes were up-regulated only in pregnancy and the CaV1.3 gene in type 1 diabetes. The results are consistent with that expression of ORAI, STIM and CaV genes correlate with a shift in immunological status of the individual in health, as during pregnancy, and in the autoimmune disease type 1 diabetes. Whether the changes are in general protective or in type 1 diabetes include some pathogenic components remains to be clarified.


The protein expression landscape of the Arabidopsis root.
Because proteins are the major functional components of cells, knowledge of their cellular localization is crucial to gaining an understanding of the biology of multicellular organisms. We have generated a protein expression map of the Arabidopsis root providing the identity and cell type-specific localization of nearly 2,000 proteins. Grouping proteins into functional categories revealed unique cellular functions and identified cell type-specific biomarkers. Cellular colocalization provided support for numerous protein-protein interactions. With a binary comparison, we found that RNA and protein expression profiles are weakly correlated. We then performed peak integration at cell type-specific resolution and found an improved correlation with transcriptome data using continuous values. We performed GeLC-MS/MS (in-gel tryptic digestion followed by liquid chromatography-tandem mass spectrometry) proteomic experiments on mutants with ectopic and no root hairs, providing complementary proteomic data. Finally, among our root hair-specific proteins we identified two unique regulators of root hair development.


SAS6-like protein in Plasmodium indicates that conoid-associated apical complex proteins persist in invasive stages within the mosquito vector.
The SAS6-like (SAS6L) protein, a truncated paralogue of the ubiquitous basal body/centriole protein SAS6, has been characterised recently as a flagellum protein in trypanosomatids, but associated with the conoid in apicomplexan Toxoplasma. The conoid has been suggested to derive from flagella parts, but is thought to have been lost from some apicomplexans including the malaria-causing genus Plasmodium. Presence of SAS6L in Plasmodium, therefore, suggested a possible role in flagella assembly in male gametes, the only flagellated stage. Here, we have studied the expression and role of SAS6L throughout the Plasmodium life cycle using the rodent malaria model P. berghei. Contrary to a hypothesised role in flagella, SAS6L was absent during gamete flagellum formation. Instead, SAS6L was restricted to the apical complex in ookinetes and sporozoites, the extracellular invasive stages that develop within the mosquito vector. In these stages SAS6L forms an apical ring, as we show is also the case in Toxoplasma tachyzoites. The SAS6L ring was not apparent in blood-stage invasive merozoites, indicating that the apical complex is differentiated between the different invasive forms. Overall this study indicates that a conoid-associated apical complex protein and ring structure is persistent in Plasmodium in a stage-specific manner.


A hemolysin associated with the mumps virus.
1. A factor capable of causing the hemolysis of the erythrocytes of man, chicken, and sheep occurs in the amniotic and allantoic fluids of chick embryos infected with the virus of mumps. 2. The hemolysin has not been found in normal fluids or in those infected with PR8 or Lee B strains of influenza virus. 3. The hemolysin is definitely inhibited by the serum of man and monkey convalescent from mumps, but only slightly by the serum of the acute phase. 4. The hemolysin is almost completely inactivated at 50 degrees C. after 10 minutes. It exhibits maximal activity at 37 degrees C. and is completely inactive at 4 degrees C. A pH range from about 7.0 to 8.0 allows for maximum activity. 5. Adsorption and elution of the hemolysin with red blood cells has been demonstrated. After elution of the hemolysin, the red blood cells exhibit an increased osmotic fragility. Similar treatment of red cells with influenza virus did not alter this property. 6. The relationship of the hemolysin to the hemagglutinin and the enzyme-like behavior of the former have been discussed.


Patterns of Hepatosplenic Brucella Abscesses on Cross-Sectional Imaging: A Review of Clinical and Imaging Features.
While diffuse involvement of liver and spleen is frequently seen in brucellosis, suppurative abscesses caused by Brucella are less common but well described. With the increased availability of cross-sectional imaging techniques, reports have become more frequent. Four patients with hepatosplenic abscesses caused by Brucella spp. are described and included in a review of 115 previously published cases. Clinical characteristics and patterns on ultrasound (US) and computed tomography imaging were analyzed. Furthermore, the proportion of patients with brucellosis affected by suppurative hepatosplenic lesions was estimated. Hepatosplenic abscesses were seen in 1.2% of patients with brucellosis and were mostly caused by Brucella melitensis. Imaging analysis revealed two main distinct patterns. Solitary abscesses involving liver more frequently than spleen, and showing characteristic central calcifications, characterize the first pattern. Multiple smaller abscesses, frequent spleen involvement, and absence of calcifications characterize the second pattern. Blood and aspirate cultures were frequently negative, however, the positivity rate increased over the past years. Indirect Coombs test was positive in 96%. Half of the patients were cured by antibiotic treatment; case fatality in this series was 1.9%. Hepatosplenic abscesses due to Brucella infections have characteristic imaging findings. Clinicians should be aware of these and the proactive use of cross-sectional imaging, particularly US, should be encouraged in endemic regions.


Altered metabolic pathways in clear cell renal cell carcinoma: A meta-analysis and validation study focused on the deregulated genes and their associated networks.
Clear cell renal cell carcinoma (ccRCC) is the predominant subtype of renal cell carcinoma (RCC). It is one of the most therapy-resistant carcinomas, responding very poorly or not at all to radiotherapy, hormonal therapy and chemotherapy. A more comprehensive understanding of the deregulated pathways in ccRCC can lead to the development of new therapies and prognostic markers. We performed a meta- analysis of 5 publicly available gene expression datasets and identified a list of co- deregulated genes, for which we performed extensive bioinformatic analysis coupled with experimental validation on the mRNA level. Gene ontology enrichment showed that many proteins are involved in response to hypoxia/oxygen levels and positive regulation of the VEGFR signaling pathway. KEGG analysis revealed that metabolic pathways are mostly altered in ccRCC. Similarly, Ingenuity Pathway Analysis showed that the antigen presentation, inositol metabolism, pentose phosphate, glycolysis/gluconeogenesis and fructose/mannose metabolism pathways are altered in the disease. Cellular growth, proliferation and carbohydrate metabolism, were among the top molecular and cellular functions of the co-deregulated genes. qRT-PCR validated the deregulated expression of several genes in Caki-2 and ACHN cell lines and in a cohort of ccRCC tissues. NNMT and NR3C1 increased expression was evident in ccRCC biopsies from patients using immunohistochemistry. ROC curves evaluated the diagnostic performance of the top deregulated genes in each dataset. We show that metabolic pathways are mostly deregulated in ccRCC and we highlight those being most responsible in its formation. We suggest that these genes are candidate predictive markers of the disease.


High dose-per-pulse electron beam dosimetry - A model to correct for the ion recombination in the Advanced Markus ionization chamber.
The purpose of this work was to establish an empirical model of the ion recombination in the Advanced Markus ionization chamber for measurements in high dose rate/dose-per-pulse electron beams. In addition, we compared the observed ion recombination to calculations using the standard Boag two-voltage-analysis method, the more general theoretical Boag models, and the semiempirical general equation presented by Burns and McEwen. Two independent methods were used to investigate the ion recombination: (a) Varying the grid tension of the linear accelerator (linac) gun (controls the linac output) and measuring the relative effect the grid tension has on the chamber response at different source-to-surface distances (SSD). (b) Performing simultaneous dose measurements and comparing the dose-response, in beams with varying dose rate/dose-per-pulse, with the chamber together with dose rate/dose-per-pulse independent Gafchromic™ EBT3 film. Three individual Advanced Markus chambers were used for the measurements with both methods. All measurements were performed in electron beams with varying mean dose rate, dose rate within pulse, and dose-per-pulse (10-2 ≤ mean dose rate ≤ 103 Gy/s, 102 ≤ mean dose rate within pulse ≤ 107 Gy/s, 10-4 ≤ dose-per-pulse ≤ 101 Gy), which was achieved by independently varying the linac gun grid tension, and the SSD. The results demonstrate how the ion collection efficiency of the chamber decreased as the dose-per-pulse increased, and that the ion recombination was dependent on the dose-per-pulse rather than the dose rate, a behavior predicted by Boag theory. The general theoretical Boag models agreed well with the data over the entire investigated dose-per-pulse range, but only for a low polarizing chamber voltage (50 V). However, the two-voltage-analysis method and the Burns & McEwen equation only agreed with the data at low dose-per-pulse values (≤ 10-2 and ≤ 10-1 Gy, respectively). An empirical model of the ion recombination in the chamber was found by fitting a logistic function to the data. The ion collection efficiency of the Advanced Markus ionization chamber decreases for measurements in electron beams with increasingly higher dose-per-pulse. However, this chamber is still functional for dose measurements in beams with dose-per-pulse values up toward and above 10 Gy, if the ion recombination is taken into account. Our results show that existing models give a less-than-accurate description of the observed ion recombination. This motivates the use of the presented empirical model for measurements with the Advanced Markus chamber in high dose-per-pulse electron beams, as it enables accurate absorbed dose measurements (uncertainty estimation: 2.8-4.0%, k = 1). The model depends on the dose-per-pulse in the beam, and it is also influenced by the polarizing chamber voltage, with increasing ion recombination with a lowering of the voltage.


Phage infection, transfection and transformation of Mycobacterium avium complex and Mycobacterium paratuberculosis.
Mycobacterium avium complex strains and Mycobacterium paratuberculosis are closely related intracellular pathogens affecting humans and animals. M. avium complex infections are a leading cause of morbidity and mortality in AIDS patients, and M. paratuberculosis is the agent of Johne's disease in ruminants. Genetic manipulation of these micro-organisms would facilitate the understanding of their pathogenesis, the construction of attenuated vaccine strains and the development of new drugs and treatment methods. This paper describes the replication of mycobacterial shuttle phasmids and plasmids, and the expression of the firefly luciferase reporter gene in M. avium complex and M. paratuberculosis. The mycobacteriophage TM4 propagated on M. smegmatis or M. paratuberculosis plaqued at the same efficiency on these two mycobacterial hosts. Screening of M. avium complex and M. paratuberculosis clinical isolates with TM4-derived luciferase reporter phages demonstrated that the majority of these isolates were susceptible to TM4. Conditions for introduction of DNA were determined by transfection of M. paratuberculosis with TM4 DNA and applied to isolate kanamycin-resistant transformants of M. avium complex and M. paratuberculosis with Escherichia coli-Mycobacterium shuttle plasmids. Recombinant plasmids were recovered from transformants without apparent loss of DNA sequences. These results provide the basis for the genetic manipulation of these pathogenic mycobacterial species.


Competency to stand trial in preadjudicatory juveniles and adults.
This study compared the competency to stand trial (CST) of 108 juveniles (ages 7 to 16 years) and 145 adults (17 years or older) undergoing pretrial, court-ordered forensic psychiatric evaluations. Adults were superior on both global and specific competency abilities (p < .001). Clustered by age (<13, 13 and 14, 15 and 16 years old), preteens accused of crimes failed to meet a Dusky v. United States standard, while 13- and 14-year-olds displayed an equal mix of abilities and deficits. Mid-adolescents (ages 15 and 16) were equivalent to adults in CST abilities except in their knowledge about plea bargaining elements. The ramifications of CST in felony juvenile transfer to adult court as well as the needs for ecologically valid, empirically based CST research on adolescents is discussed.


The energetic cost of copulation in a polygynandrous millipede
We determined the energetic cost of copulation in both sexes of Alloporus uncinatus, a polygynandrous millipede in which copulation can last for several hours and involves physical restraint of the female by the male. We found an elevation in the rate of oxygen consumption by males and a depression in the rate of oxygen consumption by females during copulation. The rate of oxygen consumption of females, but not of males, was elevated immediately after copulation, indicating that females had built up and were repaying an oxygen debt. On this basis, we estimated energy expenditure during copulation to be 30 % above resting levels in males and 14 % above resting levels in females. As matings are frequent in this species in the wild throughout the 4 month breeding season, we suggest that the energetic demands of copulation are a previously overlooked potentially significant cost of reproduction in this species and in other invertebrate species with similar mating patterns.


Assessment of Chronic Illness Care with the German version of the ACIC in different primary care settings in Switzerland.
In Switzerland the extent to which patients with chronic illnesses receive care congruent with the Chronic Care Model (CCM) is unknown. According to guidelines we translated the Assessment of Chronic Illness Care (ACIC) into German (G-ACIC). We tested the instrument in different primary care settings and compared subscales with the original testing. Difficulties encountered during the translation process consisted in the difference of health care settings in Switzerland and USA. However initial testing showed the G-ACIC to be a suitable instrument. The average ACIC subscale scores in Swiss managed care (MC)-, group (GP)- and single handed practices (SP) were higher for MC practices than for group- and single handed practices: Organization of the healthcare delivery system: MC mean (m) = 6.80 (SD 1.55), GP m = 5.42 (SD 0.99), SP m = 4.60 (SD 2.07); community linkages: MC m = 4.19 (SD 1.47), GP m = 4.83 (SD 1.81), SP m = 3.10 (SD 2.12); self-management support: MC m = 4.96 (SD 1.13), GP m = 4.73 (SD 1.40), SP m = 4.43 (SD 1.34); decision support: MC m = 4.75 (SD 1.06); GP m = 4.20 (SD 0.87), SP m = 3.25 (SD 1.59); delivery system design: MC m = 5.98 (SD 1.61), GP m = 5.05 (SD 2.05), SP m = 3.86 (SD 1.51) and clinical information systems: MC m = 4.34 (SD = 2.49), GP m = 2.06 (SD 1.35), SP m = 3.20 (SD 1.57). The G-ACIC is applicable and useful for comparing different health care settings in German speaking countries. Managed care organizations seem to implement the different components of the CCM in a greater extend than group and single handed practices. However, much room exists for further improvement.


Genetic Counselors' and Genetic Counseling Students' Attitudes Around the Clinical Doctorate and Other Advanced Educational Options for Genetic Counselors: A Report from the Genetic Counseling Advanced Degree Task Force.
Since its establishment over 40 years ago, the genetic counseling profession has grown to an estimated ~4,000 professionals in North America. While the profession has maintained the Master's degree as the entry-level and terminal degree, many other allied health professions have added advanced training pathways, such as the clinical doctorate (ClinD) either as an optional post-professional degree or required entry-level degree. Discussions regarding advanced degrees have also occurred within the genetic counseling profession, dating back to as early as the 1980s. In 2011, the Genetic Counseling Advanced Degree Task Force (GCADTF) was convened to explore the issue again, with the goal of "[engaging] all of the professional leadership organizations in the field of genetic counseling in a decision-making process about whether the profession should move to a Clinical Doctorate". As part of their work, the GCADTF surveyed practicing genetic counselors (n = 4,321) and genetic counseling students (n = 522) in the US and Canada regarding their interest in moving to the ClinD as the entry-level degree. This survey also included questions about other options for advanced training to generate data to inform future discussions around this very important professional issue. Herein, we describe the results of the survey, with particular attention to genetic counselor preferences for additional advanced education/certification opportunities and recommendations for future discussion.


Biguanides and NIDDM.
The main biguanides, metformin and phenformin, were introduced in 1957 as oral glucose-lowering agents to treat non-insulin-dependent diabetes mellitus (NIDDM). Phenformin was withdrawn in many countries because of an association with lactic acidosis, but metformin does not have the same risk if appropriately prescribed. Metformin is now widely used as a monotherapy and in combination with a sulfonylurea. Unlike sulfonylureas, metformin is not bound to plasma proteins, is not metabolized, and is eliminated rapidly by the kidney. The glucose-lowering effect occurs without stimulation of insulin secretion and results mainly from increased glucose utilization. The presence of insulin is required, and enhancement of insulin action at the postreceptor level occurs in peripheral tissues such as muscle. In peripheral tissues metformin increases insulin-mediated glucose uptake and oxidative metabolism. Metformin also increases glucose utilization by the intestine, primarily via nonoxidative metabolism. The extra lactate produced is largely extracted by the liver and serves as a substrate to sustain gluconeogenesis. This limits the extent to which metformin reduces hepatic glucose production but provides a safeguard against excessive glucose lowering. Because metformin does not cause clinical hypoglycemia, it is actually an antihyperglycemic drug. It does not cause weight gain, it helps combat hypertriglyceridemia, and it has been ascribed some vasoprotective properties. Metformin offers a useful treatment for insulin-resistant overweight NIDDM patients.


Microvascular studies on the origins of perfusion-limited hypoxia.
Two forms of hypoxia are thought to exist in tumours: (1) hypoxia caused by limitations of its diffusion (chronic hypoxia); and (2) hypoxia caused by changes in perfusion (acute hypoxia). Indirect information suggests the existence of perfusion-limited hypoxia, but there is no direct proof that fluctuations in blood flow can lead to hypoxia, nor is there any information regarding potential causes of fluctuant flow. In this study, we have begun to explore these questions using R3230AC tumours transplanted into rat dorsal-flap window chambers. Two types of fluctuant flow have been observed. The first type, usually confined to single vessels, is typified by instability of flow magnitude and direction, and total vascular stasis occurs, but only for a few seconds at a time (4% incidence). The second type of fluctuation occurs in groups of vessels and is cyclic, with cycle times ranging from 20-60 min. Total vascular stasis does not necessarily occur, but the fluctuations in red cell flux are accompanied by changes in vascular oxygen content, as measured by microelectrodes. Another source of chronic hypoxia has also been identified in these experiments. Nine per cent (9%) of vessels examined had plasma flow, but very low or absent red cell flux over periods of many minutes.


Use of dietary supplements by cardiologists, dermatologists and orthopedists: report of a survey.
Dietary supplements are regularly used by a majority of the American population, and usage by health professionals is also common. There is considerable interest in usage patterns within the population and in the reasons for using dietary supplements. The "Life...supplemented" Healthcare Professionals 2008 Impact Study (HCP Impact Study) surveyed usage of dietary supplements by physicians in three specialties: cardiology, dermatology, and orthopedics. The HCP Impact Study was conducted online by Ipsos Public Affairs for the Council for Responsible Nutrition (CRN), a trade association of the dietary supplement industry. Respondents were 900 physicians, including 300 each from three specialties--cardiology, dermatology, and orthopedics. Fifty-seven percent of cardiologists said they use dietary supplements at least occasionally, as did 75% of dermatologists and 73% of orthopedists. The product most commonly reported to be used was a multivitamin, but over 25% in each specialty said they used omega-3 fatty acids and over 20% said they used some botanical supplements. Regular dietary supplement use was reported by 37% of cardiologists, 59% of dermatologists, and 50% of orthopedists. Seventy-two percent of cardiologists, 66% of dermatologists, and 91% of orthopedists reported recommending dietary supplements to their patients. The primary reason given for recommending dietary supplements to patients was for heart health or lowering cholesterol for the cardiologists; benefits for skin, hair and nails for the dermatologists; and bone and joint health for the orthopedists. Reported dietary supplement use was relatively common in this sample of physicians, and when they recommended dietary supplements to patients, they tended to do so for reasons related to their specialty.


Neonatal head and torso vibration exposure during inter-hospital transfer.
Inter-hospital transport of premature infants is increasingly common, given the centralisation of neonatal intensive care. However, it is known to be associated with anomalously increased morbidity, most notably brain injury, and with increased mortality from multifactorial causes. Surprisingly, there have been relatively few previous studies investigating the levels of mechanical shock and vibration hazard present during this vehicular transport pathway. Using a custom inertial datalogger, and analysis software, we quantify vibration and linear head acceleration. Mounting multiple inertial sensing units on the forehead and torso of neonatal patients and a preterm manikin, and on the chassis of transport incubators over the duration of inter-site transfers, we find that the resonant frequency of the mattress and harness system currently used to secure neonates inside incubators is [Formula: see text]. This couples to vehicle chassis vibration, increasing vibration exposure to the neonate. The vibration exposure per journey (A(8) using the ISO 2631 standard) was at least 20% of the action point value of current European Union regulations over all 12 neonatal transports studied, reaching 70% in two cases. Direct injury risk from linear head acceleration (HIC15) was negligible. Although the overall hazard was similar, vibration isolation differed substantially between sponge and air mattresses, with a manikin. Using a Global Positioning System datalogger alongside inertial sensors, vibration increased with vehicle speed only above 60 km/h. These preliminary findings suggest there is scope to engineer better systems for transferring sick infants, thus potentially improving their outcomes.


Modulation of protein tyrosine phosphorylation by the extracellular matrix.
Fibronectin (FN) cross-linked to fibrin following injury provides the provisional matrix required for cells to begin tissue repair. Our previous work has demonstrated that fibroblasts adherent to multimeric FN within the context of a fibrin matrix (FN-fibrin) exhibit clear phenotypic differences from those adherent to a dimeric FN-coated surface. We hypothesize that this response to multimeric FN may be mediated by altered protein tyrosine phosphatase activity following integrin activation. NIH 3T3 cells were plated in the presence or absence of pervanadate (PV), a phosphotyrosine phosphatase inhibitor, on wells coated with FN or FN-fibrin matrix. Spread cell areas were measured after increasing incubation times and are recorded as mean cell area (mm2) +/- SEM. Alternatively, cells were lysed and equal amounts of protein were analyzed by immunoblot using a monoclonal antibody specific for phosphotyrosine. PV significantly inhibited cell spreading on FN-fibrin matrices. In contrast, PV treatment had little effect on cell area on FN alone. Analysis of cell lysates revealed that protein tyrosine phosphorylation events differ in a substrate-dependent manner. Cell attachment to a FN-fibrin matrix induces distinct cell shape and cytoskeletal organization. Inactivation of tyrosine-specific phosphatases enhances this distinction and inhibits the spreading of cells attached to this substrate. The phosphotyrosyl protein content of treated cells on FN-fibrin matrix is also diminished. These results suggest that cell-extracellular matrix interactions affect the tyrosine phosphorylation balance of the cell, thus modifying cytoskeletal organization and related signaling events.


Functional response to cholinesterase inhibitor therapy in a naturalistic Alzheimer's disease cohort.
Activities of daily living (ADL) are an essential part of the diagnostic criteria for Alzheimer's disease (AD). A decline in ADL affects independent living and has a strong negative impact on caregiver burden. Functional response to cholinesterase inhibitor (ChEI) treatment and factors that might influence this response in naturalistic AD patients need investigating. The aim of this study was to identify the socio-demographic and clinical factors that affect the functional response after 6 months of ChEI therapy. This prospective, non-randomised, multicentre study in a routine clinical setting included 784 AD patients treated with donepezil, rivastigmine or galantamine. At baseline and after 6 months of treatment, patients were assessed using several rating scales, including the Instrumental Activities of Daily Living (IADL) scale, Physical Self-Maintenance Scale (PSMS) and Mini-Mental State Examination (MMSE). Demographic and clinical characteristics were investigated at baseline. The functional response and the relationships of potential predictors were analysed using general linear models. After 6 months of ChEI treatment, 49% and 74% of patients showed improvement/no change in IADL and in PSMS score, respectively. The improved/unchanged patients exhibited better cognitive status at baseline; regarding improved/unchanged PSMS, patients were younger and used fewer anti-depressants. A more positive functional response to ChEI was observed in younger individuals or among those having the interaction effect of better preserved cognition and lower ADL ability. Patients with fewer concomitant medications or those using NSAIDs/acetylsalicylic acid showed a better PSMS response. Critical characteristics that may influence the functional response to ChEI in AD were identified. Some predictors differed from those previously shown to affect cognitive response, e.g., lower cognitive ability and older age predicted better cognitive but worse functional response.


The efficacy of clinic-based interventions aimed at increasing screening for bacterial sexually transmitted infections among men who have sex with men: a systematic review.
In many countries, the prevalence of bacterial sexually transmitted infections (STIs) among men who have sex with men (MSM) is high. We undertook a systematic review to identify clinic-based strategies for increasing screening and detection of bacterial STIs among MSM. We reviewed studies that compared screening for or detection of gonorrhea, chlamydia, and syphilis in the presence and the absence of an intervention. The primary end points were STI screening, rescreening, or detection rates. Of 1809 studies identified, 8 fulfilled the inclusion criteria; of these, 4 studies demonstrated significant increases in screening rates for gonorrhea and chlamydia using different strategies (odds ratio range, 1.4-1.9). These included the following: use of a computer alert on an electronic medical record; the introduction of clinic guidelines on STI screening; and short text messaging reminders for repeat STI screening. A further 4 studies demonstrated increases in syphilis testing (odds ratio range, 2.3-21.4), with increased detection of asymptomatic early syphilis in 2 studies. Strategies used included regular serological screening for syphilis during routine human immunodeficiency virus (HIV) care, syphilis serology included with blood tests performed as part of HIV monitoring, use of a computer alert on an electronic medical record, and an electronic medical record system to enhance syphilis retesting after syphilis treatment. A range of interventions has been used, including the application of newer technologies targeting clinicians and patients that appear to be efficacious at increasing screening of MSM for bacterial STIs. Wider application of such interventions could improve STI screening and control in this high-risk population.


Basic residues in the T-cell receptor ζ cytoplasmic domain mediate membrane association and modulate signaling.
The T-cell receptor (TCR) consists of a TCRαβ heterodimer, a TCRζ homodimer, and CD3γε and CD3δε heterodimers. The precise mechanism of T-cell triggering following TCR ligand engagement remains elusive. Previous studies reported that the cytoplasmic tail of CD3ε binds to the plasma membrane through a basic residue-rich stretch (BRS) and proposed that dissociation from the membrane is required for phosphorylation thereof. In this report we show that BRS motifs within the cytoplasmic tail of TCRζ mediate association with the plasma membrane and that TCR engagement results in TCRζ dissociation from the membrane. This dissociation requires phosphorylation of the TCRζ immunoreceptor tyrosine-based activation motifs by lymphocyte cell-specificprotein tyrosine kinase (Lck) but not ζ-chain-associated protein kinase 70 binding. Mutations of the TCRζ BRS motifs that disrupt this membrane association attenuate proximal and distal responses induced by TCR engagement. These mutations appear to alter the localization of TCRζ with respect to Lck as well as the mobility of the TCR complex. This study reveals that tyrosine phosphorylation of the TCRζ cytoplasmic domain regulates its association with the plasma membrane and highlights the functional importance of TCRζ BRS motifs.


Effects of the gay publishing boom on classes of titles retrieved under the subject headings "Homosexuality," "Gay Men," and "Gays" in the OCLC WorldCat database.
What do searchers find when they look for literature on homosexuality? This question has profound implications for older as well as younger gays in their coming out, as well as in their subsequent identity development. Library records provide credible data to answer the question, since they represent relatively free sources of information, unlike data from bookstores, publishers, and some World Wide Web sites. The records of WorldCat, the world's largest union database of library records, comprise over 30 million records listed in the Online Computer Library Center. For the purposes of the study, 18,757 records listed under "Homosexuality," "Gay Men," and "Gays" were downloaded; records for "Lesbian" and "Lesbians" were not examined. Findings of the study suggest that while there has indeed been considerable growth in terms of the quantity of gay literature produced since 1969, such gains may be offset by the deteriorating quality of cataloging copy, which makes the experience of browsing records a discouraging and confusing one.


Parameters affecting the fusion of unilamellar phospholipid vesicles with planar bilayer membranes.
It was previously shown (Cohen, F. S., J. Zimmerberg, and A. Finkelstein, 1980, J. Gen. Physiol., 75:251-270) that multilamellar phospholipid vesicles can fuse with decane-containing phospholipid bilayer membranes. An essential requirement for fusion was an osmotic gradient across the planar membrane, with the vesicle-containing (cis) side hyperosmotic with respect to the opposite (trans) side. We now report that unilamellar vesicles will fuse with "hydrocarbon-free" membranes subject to these same osmotic conditions. Thus the same conditions that apply to fusion of multilamellar vesicles with planar bilayer membranes also apply to fusion of unilamellar vesicles with these membranes, and hydrocarbon is not required for the fusion process. If the vesicles and/or planar membrane contain negatively charged lipids, divalent cation (approximately 15 mM Ca++) is required in the cis compartment (in addition to the osmotic gradient across the membrane) to obtain substantial fusion rates. On the other hand, vesicles made from uncharged lipids readily fuse with planar phosphatidylethanolamine planar membranes in the near absence of divalent cation with just an osmotic gradient. Vesicles fuse much more readily with phosphatidylethanolamine-containing than with phosphatidylcholine-containing planar membranes. Although hydrocarbon (decane) is not required in the planar membrane for fusion, it does affect the rate of fusion and causes the fusion process to be dependent on stirring in the cis compartment.


Effects of Meditation and Mind-Body Exercises on Older Adults' Cognitive Performance: A Meta-analysis.
Meditation and mind-body exercises are suggested to delay decline or enhance cognitive capabilities in older adults. However, their effectiveness remains uncertain. This study assessed the effectiveness of meditation and mind-body exercises to improve cognition in elderly people aged 60 years or above. Moderator variables were also explored. A databases search (MEDLINE, EMBASE, CINAHL, PsycINFO, Cochrane Library, Web of Science, CNKI, and Wangfang) was conducted from the first available date to January 10, 2018. Inclusion criteria include (a) human older adults aged 60 years or above, (b) meditation, Tai Chi, Qigong, or yoga intervention, (c) intervention should be structured, (d) inclusion of a control group, (e) at least one outcome measure of cognition was measured at baseline and post-training, and (f) peer-reviewed journal articles in English or Chinese. Forty-one studies (N = 3,551) were included in the meta-analysis. In general, meditation and mind-body exercises improve cognition in the elderly people (SMD = 0.34, 95% CI: 0.19 to 0.48), but the cognition-enhancing effects depend on the type of exercise. In addition, cognitive performance is only improved when the length of intervention is longer than 12 weeks, exercise frequency is 3-7 times/week, or duration of an exercise session is 45-60 min/session. This study suggests that meditation and mind-body exercises are effective to improve cognition of older adults aged 60 years or above, and exercise parameters should be considered for intervention planning.


Euglycemic agent-mediated hypothalamic transcriptomic manipulation in the N171-82Q model of Huntington disease is related to their physiological efficacy.
Our aim was to employ novel analytical methods to investigate the therapeutic treatment of the energy regulation dysfunction occurring in a Huntington disease (HD) mouse model. HD is a neurodegenerative disorder that is characterized by progressive motor impairment and cognitive alterations. Changes in neuroendocrine function, body weight, energy metabolism, euglycemia, appetite function, and gut function can also occur. It is likely that the locus of these alterations is the hypothalamus. We determined the effects of three different euglycemic agents on HD progression using standard physiological and transcriptomic signature analyses. N171-82Q HD mice were treated with insulin, Exendin-4, and the newly developed GLP-1-Tf to determine whether these agents could improve energy regulation and delay disease progression. Blood glucose, insulin, metabolic hormone levels, and pancreatic morphology were assessed. Hypothalamic gene transcription, motor coordination, and life span were also determined. The N171-82Q mice exhibited significant alterations in hypothalamic gene transcription signatures and energy metabolism that were ameliorated, to varying degrees, by the different euglycemic agents. Exendin-4 or GLP-1-Tf (but not insulin) treatment also improved pancreatic morphology, motor coordination, and increased life span. Using hypothalamic transcription signature analyses, we found that the physiological efficacy variation of the drugs was evident in the degree of reversal of the hypothalamic HD pathological signature. Euglycemic agents targeting hypothalamic and energy regulation dysfunction in HD could potentially alter disease progression and improve quality of life in HD.


Glucose-6-phosphate dehydrogenase and leptin are related to marbling differences among Limousin and Angus or Japanese Black x Angus steers.
This work investigated the metabolic basis for the variability of carcass and i.m. adiposity in cattle. Our hypothesis was that the comparison of extreme breeds for adiposity might allow for the identification of some metabolic pathways determinant for carcass and i.m. adiposity. Thus, 23- to 28-mo-old steers of 3 breeds, 2 with high [Angus or Japanese Black x Angus (J. Black cross)] and 1 with low (Limousin) i.m. and carcass adiposity, were used to measure activities or mRNA levels, or both, of enzymes involved in de novo lipogenesis [acetyl-coA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme], circulating triacylglycerol (TAG) uptake (lipoprotein lipase), and fatty acid esterification (glycerol-3-phosphate dehydrogenase), as well as the mRNA level of leptin, an adiposity-related factor. In a first study, enzyme activities were assayed in the s.c. adipose tissue (AT), the oxidative rectus abdominis, and the glycolytic semitendinosus muscles from steers finished for 6 mo. Compared with Angus or J. Black cross, Limousin steers had a 27% less (P = 0.003) rib fat thickness, and 23 and 29% less (P < or = 0.02) FAS and G6PDH activities in s.c. AT. In rectus abdominis and semitendinosus, the 75% less (P < 0.001) TAG content was concomitant with 50% less (P < 0.001) G6PDH activity. In a second study, enzyme activities plus mRNA levels were assayed in an oxido-glycolytic muscle, the longissimus thoracis (LT), in the i.m. AT dissected from LT, and in s.c. AT from the same Limousin steers and from Angus steers finished for 10 mo. Compared with Angus, the 50% less (P < 0.001) rib fat thickness in Limousin contrasted with the 1.1- to 5.8-fold greater (P < or = 0.02) mRNA levels or activities, or both, of acetyl-coA carboxylase, G6PDH, lipoprotein lipase, and glycerol-3-phosphate dehydrogenase in s.c. AT. Conversely, the 90% less (P < 0.001) TAG content in Limousin LT was concomitant to the 79 and 83% less (P < or = 0.002) G6PDH activity and leptin mRNA level. Such differences could arise from a greater number of adipocytes in LT from Angus steers because no difference was found between Limousin and Angus for G6PDH activity and leptin mRNA in i.m. AT. We conclude that FAS and G6PDH in s.c. AT could be involved in differences in carcass adiposity, but this relationship disappeared when the fatness increased strongly. Leptin and G6PDH are related to the expression of marbling whatever the body condition and thus could be relevant indicators of marbling in beef cattle.


Alginate-encapsulated HepG2 cells in a fluidized bed bioreactor maintain function in human liver failure plasma.
Alginate-encapsulated HepG2 cells cultured in microgravity have the potential to serve as the cellular component of a bioartificial liver. This study investigates their performance in normal and liver failure (LF) human plasma over 6-8 h in a fluidized bed bioreactor. After 8 days of microgravity culture, beads containing 1.5 x 10(9) cells were perfused for up to 8 h at 48 mL/min with 300 mL of plasma. After exposure to 90% LF plasma, vital dye staining showed maintained cell viability, while a 7% increase in lactate dehydrogenase activity indicated minimal cell damage. Glucose consumption, lactate production, and a 4.3-fold linear increase in alpha-fetoprotein levels were observed. Detoxificatory function was demonstrated by quantification of bilirubin conjugation, urea synthesis, and Cyp450 1A activity. These data show that in LF plasma, alginate-encapsulated HepG2 cells can maintain viability, and metabolic, synthetic, and detoxificatory activities, indicating that the system can be scaled-up to form the biological component of a bioartificial liver.


Esthetic outcome of implant-supported single-tooth replacements assessed by the patient and by prosthodontists.
The objective of this study was to assess and compare patients' and clinicians' judgments of the esthetic outcome of implant-supported single-tooth replacements. In all, 29 patients with 41 single implant-supported crowns in the maxillary anterior region participated in the study. The esthetic outcome of the implant-supported crowns was assessed by the patients and by 5 prosthodontists by means of a questionnaire in which various esthetics-related variables were addressed and responded to using visual analogue scales. Multiple regression analyses were used to evaluate the influence of the variables on the "overall satisfaction" with the implant-supported crown. Most variables in the patients' assessments revealed mean values above 90% and median values close to 100%. No single factor used in the multiple regression analysis was found to influence a patient's satisfaction with appearance of the crown at a statistically significant level. The clinicians' degree of satisfaction was for all variables lower than that of the patients. In 89% of the cases the clinicians could correctly locate the single implant-supported crown. Among the variables assessed, surrounding soft tissue appearance and form of the crown had the strongest influence on the clinician's overall satisfaction with the appearance of the crown. Appreciation of the esthetic outcome of the single implant-supported crowns was rated higher by the patients than by the prosthodontists. Furthermore, factors considered by professionals to be of significance for the esthetic result of the restorative treatment may not be of decisive importance for the patient's satisfaction.


Does medicine run in the family-evidence from three generations of physicians in Sweden: retrospective observational study.
To examine occupational heritability in medicine and changes in heritability over time, with Swedish population wide administrative data that allowed mapping family trees of physicians spanning up to three generations. Retrospective observational study. Individual level administrative registry data from Sweden. Physicians born in 1950-90 and living in Sweden at some time during 2001-16 (n=47 400). The proportion of individuals with a completed medical degree with at least one parent who also trained in medicine, and the change in this proportion across birth cohorts. Additional analyses were conducted among other relatives (grandparents, aunts and uncles, and siblings) and for individuals with a law degree. For 27 788 physicians, where the educational background for both parents was known, 14% had a parent who was also a physician and 2% had two parents who were physicians. The proportion of physicians with at least one physician parent increased significantly over time, from 6% for physicians born in 1950-59 to 20% for physicians born in 1980-90 (P<0.001). The same pattern of increasing occupational heritability was not seen for individuals with law degrees. In recent cohorts of physicians in Sweden, one in five had a parent who was also a physician, more than triple the proportion seen for physicians born three decades earlier. A similar pattern was not seen in lawyers, suggesting that increasing occupational heritability in medicine does not reflect intergenerational persistence of high paying degrees alone. Rather, for physicians in Sweden, medicine might increasingly run in families.


Effect of all-trans retinoic acid on newly diagnosed acute promyelocytic leukemia patients: results of a Brazilian center.
Thirty-seven patients with acute promyelocytic leukemia (APL) were treated with all-trans retinoic acid (ATRA). Patients received 45 mg m-2 day-1 po of ATRA until complete remission (CR) was achieved, defined as: a) presence of less than 5% blasts in the bone marrow, with b) white blood cells > 10(3)/mm3, c) platelets > 10(5)/mm3 and d) hemoglobin concentration > 8 g/dl, with no blood or platelet transfusions. Thirty-one (83.7%) patients achieved CR by day 50, and 75% of these before day 30. Correction of the coagulopathy, achieved between days 2 and 10 (mean, 3 days), was the first evidence of response to treatment. Only one patient had been previously treated with chemotherapy and three had the microgranular variant M3 form. Dryness of skin and mucosae was the most common side effect observed in 82% of the patients. Thrombosis, hepatotoxicity and retinoid acid syndrome (RAS) were observed in 7 (19%), 6 (16%) and 4 (11%) patients, respectively. Thirteen (35%) patients had to be submitted to chemotherapy due to hyperleukocytosis (above 40 x 10(3)/mm3) and six of these presented with new signs of coagulopathy after chemotherapy. Four (11%) patients died secondarily to intracerebral hemorrhage (IH) and two (5.4%) dropped out of the protocol due to severe ATRA side effects (one RAS and one hepatotoxicity). RAS and IH were related strictly to hyperleukocytosis. The reduced use of platelets and fresh frozen plasma probably lowered the total cost of treatment. We conclude that ATRA is an effective agent for inducing complete remission in APL patients.


Molecular analysis reveals heterogeneity of mouse mammary tumors conditionally mutant for Brca1.
Development of therapies for patients with BRCA1 mutations has been hampered by lack of readily available in vitro and in vivo models. We recently showed that transplantation of transgenic mammary tumors as cell suspensions into naïve recipients generates reproducible tumors with remarkable stability of gene expression profile. We examined the expression profiles of original and serially transplanted mammary tumors from Brca1 deficient mice, and tumor derived cell lines to validate their use for preclinical testing and studies of tumor biology. Original tumors, serially transplanted and multiple cell lines derived from Brca1 mammary tumors were characterized by morphology, gene and protein expression, and cell surface markers. Gene expression among Brca1 tumors showed more heterogeneity than among previously characterized tumors from MMTV-PyMT and -Wnt1 models. Gene expression data segregated Brca1 tumors into 3 distinct types: basal, mixed luminal, and tumors with epithelial-to-mesenchymal transition (EMT). Serial transplantation of individual tumors and multiple cell lines derived from the original tumors recapitulated the molecular characteristics of each tumor of origin. One tumor had distinct features of EMT and gave rise to cell lines that contained a distinct CD44+/CD24-/low population that may correlate with human breast cancer stem cells. Although individual tumors expanded by transplantation maintain the genomic profile of the original tumors, the heterogeneity among Brca1 tumors limits the extent of their use for preclinical testing. However, cell lines offer a robust material for understanding tumor biology and response to therapies driven by BRCA1 deficiency.


Modelling micro-pollutant fate in wastewater collection and treatment systems: status and challenges.
This paper provides a comprehensive summary on modelling of micro-pollutants' (MPs) fate and transport in wastewater. It indicates the motivations of MP modelling and summarises and illustrates the current status. Finally, some recommendations are provided to improve and diffuse the use of such models. In brief, we conclude that, in order to predict the contaminant removal in centralised treatment works, considering the dramatic improvement in monitoring and detecting MPs in wastewater, more mechanistic approaches should be used to complement conventional, heuristic and other fate models. This is crucial, as regional risk assessments and model-based evaluations of pollution discharge from urban areas can potentially be used by decision makers to evaluate effluent quality regulation, and assess upgrading requirements, in the future.


Current Total Knee Designs: Does Baseplate Roughness or Locking Mechanism Design Affect Polyethylene Backside Wear?
Tibial baseplate roughness and polyethylene-insert micromotion resulting from locking-mechanism loosening can lead to polyethylene backside wear in TKAs. However, many retrieval studies examining these variables have evaluated only older TKA implant designs. We used implant-retrieval analysis to examine if there were differences in: (1) backside damage scores, (2) backside damage modes, and (3) backside linear wear rates in five TKA implant designs owing to differing baseplate surface roughness and locking mechanisms. Additionally, we examined if (4) patient demographics influence backside damage and wear. Five TKA implant models (four modern and one historical design) were selected with different tibial baseplate and/or locking mechanism designs. Six tibial inserts retrieved at the time of revision from each TKA model were matched for time in vivo, age of the patient at TKA revision, BMI, sex, revision number, and revision reason. Each insert backside was analyzed for: (1) visual total damage score and (2) individual visual damage modes, both by two observers and with an intraclass correlation coefficient of 0.66 (95% CI, 0.39-0.92), and (3) linear wear rate measured by micro-CT. Median primary outcomes were compared among the five designs. For our given sample size among five groups we could detect with 80% power a 10-point difference in damage score and an 0.11-mm per year difference in wear rate. The polished tibial design with a partial peripheral capture locking mechanism and anterior constraint showed a lower total damage score compared with the nonpolished tibial design with only a complete peripheral-rim locking mechanism (median, 12.5; range, 9.5-18.0; 95% CI, 9.58-16.42 versus median, 22.3; range, 15.5-27.0; 95% CI, 17.5-26.5; p = 0.019). The polished baseplate with a tongue-in-groove locking mechanism showed more abrasions than the nonpolished baseplate with a peripheral-rim capture and antirotational island (median, 7.25; range, 0.5-8.0; 95% CI, 2.67-8.99 versus median, 0.75; range, 0-1.5; 95% CI, 0.20-1.47; p = 0.016)). Dimpling was a unique wear mode to the nonpolished baseplates with the peripheral-rim capture and antirotational island (median, 5.5; range, 2.0-9.0; 95% CI, 2.96-8.38) and the peripheral-rim capture alone (median, 9.0; range, 6.0-10.0; 95% CI, 7.29-10.38). Overall, the linear wear rate for polished designs was lower than for nonpolished designs (0.0102 ± 0.0044 mm/year versus 0.0224 ± 0.0119 mm/year; p < 0.001). Two of the polished baseplate designs, the partial peripheral capture with anterior constraint (median, 0.083 mm/year; range, 0.0037-0.0111 mm/year; 95% CI, 0.0050-0.0107 mm versus median, 0.0245 mm/year; range, 0.014-0.046 mm/year; 95% CI, 0.0130-0.0414 mm; p = 0.008) and the tongue-in-groove locking mechanism (median, 0.0085 mm/year; range, 0.005-0.015 mm/year; 95% CI, 0.0045-0.0138 mm; p = 0.032) showed lower polyethylene linear wear rates compared with the nonpolished baseplate design with only a peripheral-rim capture. Total damage scores and linear wear rates were highest involving the nonpolished design with only a peripheral rim capture. There were no differences among the other TKA designs regarding damage and wear, but this finding should be considered in the setting of a relatively small sample size. Our study showed that in the complex interplay between baseplate surface finish and locking mechanism design, a polished baseplate with a robust locking mechanism had the lowest backside damage and linear wear. However, improvements in locking mechanism design in nonpolished baseplates potentially may offset some advantages of a polished baseplate. Further retrieval analyses need to be done to confirm such findings, especially analyzing current crosslinked polyethylene. Additionally, we need mid- and long-term studies comparing TKA revisions attributable to wear and osteolysis among implants before understanding if such design differences are clinically relevant.


Assessing Exposures to Magnetic Resonance Imaging's Complex Mixture of Magnetic Fields for In Vivo, In Vitro, and Epidemiologic Studies of Health Effects for Staff and Patients.
A complex mixture of electromagnetic fields is used in magnetic resonance imaging (MRI): static, low-frequency, and radio frequency magnetic fields. Commonly, the static magnetic field ranges from one to three Tesla. The low-frequency field can reach several millitesla and with a time derivative of the order of some Tesla per second. The radiofrequency (RF) field has a magnitude in the microtesla range giving rise to specific absorption rate values of a few Watts per kilogram. Very little attention has been paid to the case where there is a combined exposure to several different fields at the same time. Some studies have shown genotoxic effects in cells after exposure to an MRI scan while others have not demonstrated any effects. A typical MRI exam includes muliple imaging sequences of varying length and intensity, to produce different types of images. Each sequence is designed with a particular purpose in mind, so one sequence can, for example, be optimized for clearly showing fat water contrast, while another is optimized for high-resolution detail. It is of the utmost importance that future experimental studies give a thorough description of the exposure they are using, and not just a statement such as "An ordinary MRI sequence was used." Even if the sequence is specified, it can differ substantially between manufacturers on, e.g., RF pulse height, width, and duty cycle. In the latest SCENIHR opinion, it is stated that there is very little information regarding the health effects of occupational exposure to MRI fields, and long-term prospective or retrospective cohort studies on workers are recommended as a high priority. They also state that MRI is increasingly used in pediatric diagnostic imaging, and a cohort study into the effects of MRI exposure on children is recommended as a high priority. For the exposure assessment in epidemiological studies, there is a clear difference between patients and staff and further work is needed on this. Studies that explore the possible differences between MRI scan sequences and compare them in terms of exposure level are warranted.


Fine-scale variation in meiotic recombination in Mimulus inferred from population shotgun sequencing.
Meiotic recombination rates can vary widely across genomes, with hotspots of intense activity interspersed among cold regions. In yeast, hotspots tend to occur in promoter regions of genes, whereas in humans and mice, hotspots are largely defined by binding sites of the positive-regulatory domain zinc finger protein 9. To investigate the detailed recombination pattern in a flowering plant, we use shotgun resequencing of a wild population of the monkeyflower Mimulus guttatus to precisely locate over 400,000 boundaries of historic crossovers or gene conversion tracts. Their distribution defines some 13,000 hotspots of varying strengths, interspersed with cold regions of undetectably low recombination. Average recombination rates peak near starts of genes and fall off sharply, exhibiting polarity. Within genes, recombination tracts are more likely to terminate in exons than in introns. The general pattern is similar to that observed in yeast, as well as in positive-regulatory domain zinc finger protein 9-knockout mice, suggesting that recombination initiation described here in Mimulus may reflect ancient and conserved eukaryotic mechanisms.


Role of the salt bridge between glutamate 546 and arginine 907 in preservation of autoinhibited form of Apaf-1.
Apaf-1, the key element of apoptotic mitochondrial pathway, normally exists in an auto-inhibited form inside the cytosol. WRD-domain of Apaf-1 has a critical role in the preservation of auto-inhibited form; however the underlying mechanism is unclear. It seems the salt bridges between WRD and NOD domains are involved in maintaining the inactive conformation of Apaf-1. At the present study, we have investigated the effect of E546-R907 salt bridge on the maintenance of auto-inhibited form of human Apaf-1. E546 is mutated to glutamine (Q) and arginine (R). Over-expression of wild type Apaf-1 and its E546Q and E546R variants in HEK293T cells does not induce apoptosis unlike - HL-60 cancer cell line. In vitro apoptosome formation assay showed that all variants are cytochrome c and dATP dependent to form apoptosome and activate endogenous procaspase-9 in Apaf-1-knockout MEF cell line. These results suggest that E546 is not a critical residue for preservation of auto-inhibited Apaf-1. Furthermore, the behavior of Apaf-1 variants for in vitro apoptosome formation in HEK293T cell is similar to exogenous wild type Apaf-1. Wild type and its variants can form apoptosome in HEK293T cell with different procaspase-3 processing pattern in the presence and absence of exogenous cytochrome c and dATP.


Designing active communities: a coordinated action framework for planners and public health professionals.
Community design can have a positive or negative influence on the physical activity level of residents. The complementary expertise of professionals from both planning and public health is needed to build active communities. The current study aimed to develop a coordinated framework for planners and public health professionals to enhance the design of active communities. Planners and public health professionals working in Ontario, Canada were recruited to participate in a concept mapping process to identify ways they should work together to enhance the design of active communities. This process generated 72 actions that represent collaborative efforts planners and public health professionals should engage in when designing active communities. These actions were then organized by importance and feasibility. This resulted in a coordinated action framework that includes 19 proximal and 6 distal coordinated actions for planners and public health professionals. Implementation of the recommended actions has the potential to make a difference in community design as a way to enhance physical activity in community members. This Coordinated Action Framework provides a way to address physical inactivity from an environmental and policy standpoint.


Biomineralization of arsenate to arsenic sulfides is greatly enhanced at mildly acidic conditions.
Arsenic (As) is an important water contaminant due to its high toxicity and widespread occurrence. Arsenic-sulfide minerals (ASM) are formed during microbial reduction of arsenate (As(V)) and sulfate (SO4(2-)). The objective of this research is to study the effect of the pH on the removal of As due to the formation of ASM in an iron-poor system. A series of batch experiments was used to study the reduction of SO4(2-) and As(V) by an anaerobic biofilm mixed culture in a range of pH conditions (6.1-7.2), using ethanol as the electron donor. Total soluble concentrations and speciation of S and As were monitored. Solid phase speciation of arsenic was characterized by x-ray adsorption spectroscopy (XAS). A marked decrease of the total aqueous concentrations of As and S was observed in the inoculated treatments amended with ethanol, but not in the non-inoculated controls, indicating that the As-removal was biologically mediated. The pH dramatically affected the extent and rate of As removal, as well as the stoichiometric composition of the precipitate. The amount of As removed was 2-fold higher and the rate of the As removal was up to 17-fold greater at pH 6.1 than at pH 7.2. Stoichiometric analysis and XAS results confirmed the precipitate was composed of a mixture of orpiment and realgar, and the proportion of orpiment in the sample increased with increasing pH. The results taken as a whole suggest that ASM formation is greatly enhanced at mildly acidic pH conditions.


Spatiotemporally Super-Resolved Volumetric Traction Force Microscopy.
Quantification of mechanical forces is a major challenge across biomedical sciences. Yet such measurements are essential to understanding the role of biomechanics in cell regulation and function. Traction force microscopy remains the most broadly applied force probing technology but typically restricts itself to single-plane two-dimensional quantifications with limited spatiotemporal resolution. Here, we introduce an enhanced force measurement technique combining 3D super-resolution fluorescence structural illumination microscopy and traction force microscopy (3D-SIM-TFM) offering increased spatiotemporal resolution, opening-up unprecedented insights into physiological three-dimensional force production in living cells.


Undertreatment, treatment trends, and treatment dissatisfaction among patients with psoriasis and psoriatic arthritis in the United States: findings from the National Psoriasis Foundation surveys, 2003-2011.
Psoriasis and psoriatic arthritis inflict significant morbidity. Data on undertreatment, treatment use, and treatment satisfaction are paramount to identify priority areas for advocacy, education, and research to improve patient outcomes. To determine the extent of nontreatment and undertreatment of psoriatic diseases, trends in treatment use, treatment satisfaction, and reasons for medication discontinuation among patients with psoriasis and psoriatic arthritis. We used the national survey data collected by the National Psoriasis Foundation via biannual surveys conducted from January 1, 2003, through December 31, 2011, in the United States. Survey data were collected from randomly sampled patients with psoriasis and psoriatic arthritis in the US population from a database of more than 76,000 patients with psoriatic diseases. Nontreatment, undertreatment, and treatment trends determined by the use of prescription medication (topical, phototherapeutic, oral systemic, and biologic), as well as treatment satisfaction and reasons for medication discontinuation. A total of 5604 patients with psoriasis or psoriatic arthritis completed the survey. From 2003 through 2011, patients who were untreated ranged from 36.6% to 49.2% of patients with mild psoriasis, 23.6% to 35.5% of patients with moderate psoriasis, and 9.4% to 29.7% of patients with severe psoriasis. Among those receiving treatment, 29.5% of patients with moderate psoriasis and 21.5% of patients with severe psoriasis were treated with topical agents alone. The most frequently used phototherapy modality is UV-B, whereas methotrexate is the most commonly used oral agent. Although adverse effects and a lack of effectiveness were primary reasons for discontinuing biological agents, the inability to obtain adequate insurance coverage was among the top reasons for discontinuation. Overall, 52.3% of patients with psoriasis and 45.5% of patients with psoriatic arthritis were dissatisfied with their treatment. Nontreatment and undertreatment of patients with psoriasis and psoriatic arthritis remain a significant problem in the United States. While various treatment modalities are available for psoriasis and psoriatic arthritis, widespread treatment dissatisfaction exists. Efforts in advocacy and education are necessary to ensure that effective treatments are accessible to this patient population.


Coral host transcriptomic states are correlated with Symbiodinium genotypes.
A mutualistic relationship between reef-building corals and endosymbiotic dinoflagellates (Symbiodinium spp.) forms the basis for the existence of coral reefs. Genotyping tools for Symbiodinium spp. have added a new level of complexity to studies concerning cnidarian growth, nutrient acquisition, and stress. For example, the response of the coral holobiont to thermal stress is connected to the host-Symbiodinium genotypic combination, as different partnerships can have different bleaching susceptibilities. In this study, we monitored Symbiodinium physiological parameters and profiled the coral host transcriptional responses in acclimated, thermally stressed, and recovered fragments of the coral Montastraea faveolata using a custom cDNA gene expression microarray. Interestingly, gene expression was more similar among samples with the same Symbiodinium content rather than the same experimental condition. In order to discount for host-genotypic effects, we sampled fragments from a single colony of M. faveolata containing different symbiont types, and found that the host transcriptomic states grouped according to Symbiodinium genotype rather than thermal stress. As the first study that links coral host transcriptomic patterns to the clade content of their Symbiodinium community, our results provide a critical step to elucidating the molecular basis of the apparent variability seen among different coral-Symbiodinium partnerships.


Coffee melanoidins: structures, mechanisms of formation and potential health impacts.
During the roasting process, coffee bean components undergo structural changes leading to the formation of melanoidins, which are defined as high molecular weight nitrogenous and brown-colored compounds. As coffee brew is one of the main sources of melanoidins in the human diet, their health implications are of great interest. In fact, several biological activities, such as antioxidant, antimicrobial, anticariogenic, anti-inflammatory, antihypertensive, and antiglycative activities, have been attributed to coffee melanoidins. To understand the potential of coffee melanoidin health benefits, it is essential to know their chemical structures. The studies undertaken to date dealing with the structural characterization of coffee melanoidins have shown that polysaccharides, proteins, and chlorogenic acids are involved in coffee melanoidin formation. However, exact structures of coffee melanoidins and mechanisms involved in their formation are far to be elucidated. This paper systematizes the available information and provides a critical overview of the knowledge obtained so far about the structure of coffee melanoidins, mechanisms of their formation, and their potential health implications.


A novel approach to reducing number of sensing units for wearable gait analysis systems.
Gait analysis methods to estimate spatiotemporal measures, based on two, three or four gyroscopes attached on lower limbs have been discussed in the literature. The most common approach to reduce the number of sensing units is to simplify the underlying biomechanical gait model. In this study, we propose a novel method based on prediction of movements of thighs from movements of shanks. Datasets from three previous studies were used. Data from the first study (ten healthy subjects and ten with Parkinson's disease) were used to develop and calibrate a system with only two gyroscopes attached on shanks. Data from two other studies (36 subjects with hip replacement, seven subjects with coxarthrosis, and eight control subjects) were used for comparison with the other methods and for assessment of error compared to a motion capture system. Results show that the error of estimation of stride length compared to motion capture with the system with four gyroscopes and our new method based on two gyroscopes was close ( -0.8 ±6.6 versus 3.8 ±6.6 cm). An alternative with three sensing units did not show better results (error: -0.2 ±8.4 cm). Finally, a fourth that also used two units but with a simpler gait model had the highest bias compared to the reference (error: -25.6 ±7.6 cm). We concluded that it is feasible to estimate movements of thighs from movements of shanks to reduce number of needed sensing units from 4 to 2 in context of ambulatory gait analysis.


How useful are Cochrane reviews in identifying research needs?
To determine the extent to which reports of Cochrane reviews recommend the need for further research and, if so, the extent to which they make suggestions regarding that research. We examined all 2535 reviews in Issue 4, 2005 of The Cochrane Library. Each review was categorized on the basis of whether a suggestion was included about specific interventions, participants, or outcome measures that should be included in future research. We also identified the frequency with which reviews conclude that no more research is needed or feasible, noted the need for further systematic reviewing, and refered to a relevant ongoing or planned study. We also report the number of studies listed in the 'Ongoing Studies' section in each review. Only 3.2% of reviews suggested explicitly that no more research is needed or feasible. In 82.0% of reviews, suggestions were made as to the specific interventions that need evaluating, in 30.2% the appropriate participants were suggested, and in 51.9% outcome measures were suggested. Suggestions for all three domains were made in 16.9% of the reviews. While 11.6% did not include a specific suggestion about any of these domains, 21.2% of reviews mention a relevant ongoing or planned study in one or both of the 'Implications for Research' and the 'Ongoing Studies' sections. Most Cochrane reviews identify residual uncertainty and are a rich source of suggestions for further health-care research.


Have preferences of girls changed almost 3 years after the much debated start of the HPV vaccination program in The Netherlands? A discrete choice experiment.
To assess how girls' preferences have changed almost 3 years after the much debated start of the human papillomavirus (HPV) vaccination program. A discrete choice experiment (DCE) was conducted among girls aged 11-15 years who were invited, or were not yet invited, to get vaccinated. A panel latent class model was used to determine girls' preferences for vaccination based on five characteristics: degree of protection against cervical cancer; duration of protection; risk of mild side-effects; age of vaccination; and the number of required doses of the vaccine. The response rate was 85% (500/592). Most girls preferred vaccination at age 14 years (instead of at age 9 years) and a 2-dose scheme (instead of the current 3-dose scheme). Girls were willing to trade-off 7% (CI: 3.2% to 10.8%) of the degree of protection to have 10% less risk of mild side-effects, and 4% (CI: 1.2% to 5.9%) to receive 2 doses instead of 3 doses. Latent class analyses showed that there was preference heterogeneity among girls, i.e., higher educated girls and HPV vaccinated girls had a higher probability to opt for HPV vaccination at a higher age than lower educated girls or non-vaccinated girls. Three years after the start of HPV vaccination program the risk of mild side-effects and age at vaccination seem to have become less important. For the Dutch national immunization program, we recommend not to lower the current target age of 12 years. A 2-dose scheme may result in a higher uptake and we recommend that if this scheme is introduced, it needs to receive adequate publicity.


Periprocedural Intravenous Heparin During Endovascular Treatment for Ischemic Stroke: Results From the MR CLEAN Registry.
Background and Purpose- Intravenous administration of heparin during endovascular treatment for ischemic stroke may improve outcomes. However, risks and benefits of this adjunctive therapy remain uncertain. We aimed to evaluate periprocedural intravenous heparin use in Dutch stroke intervention centers and to assess its efficacy and safety. Methods- Patients registered between March 2014 and June 2016 in the MR CLEAN Registry (Multicenter Randomized Clinical Trial of Endovascular Treatment of Acute Ischemic Stroke), including all patients treated with endovascular treatment in the Netherlands, were analyzed. The primary outcome was functional outcome (modified Rankin Scale) at 90 days. Secondary outcomes were successful recanalization (extended Thrombolysis in Cerebral Infarction ≥2B), symptomatic intracranial hemorrhage, and mortality at 90 days. We used multilevel regression analysis to evaluate the association of periprocedural intravenous heparin on outcomes, adjusted for center effects and prognostic factors. To account for possible unobserved confounding by indication, we analyzed the effect of center preference to administer intravenous heparin, defined as percentage of patients treated with intravenous heparin in a center, on functional outcome. Results- One thousand four hundred eighty-eight patients from 16 centers were analyzed, of whom 398 (27%) received intravenous heparin (median dose 5000 international units). There was substantial between-center variability in the proportion of patients treated with intravenous heparin (range, 0%-94%). There was no significant difference in functional outcome between patients treated with intravenous heparin and those without (adjusted common odds ratio, 1.17; 95% CI, 0.87-1.56), successful recanalization (adjusted odds ratio, 1.24; 95% CI, 0.89-1.71), symptomatic intracranial hemorrhage (adjusted odds ratio, 1.13; 95% CI, 0.65-1.99), or mortality (adjusted odds ratio, 0.95; 95% CI, 0.66-1.38). Analysis at center level showed that functional outcomes were better in centers with higher percentages of heparin administration (adjusted common odds ratio, 1.07 per 10% more heparin, 95% CI, 1.01-1.13). Conclusions- Substantial between-center variability exists in periprocedural intravenous heparin use during endovascular treatment, but the treatment is safe. Centers using heparin more often had better outcomes. A randomized trial is needed to further study these effects.