An Educational Intervention Decreases Opioid Prescribing After General Surgical Operations.
The aim of this study was to determine whether an educational intervention was sufficient to decrease opioid prescribing after general surgical operations. We recently analyzed opioid prescription and use for 5 common outpatient operations at our institution: partial mastectomy (PM), PM with sentinel lymph node biopsy (PM SLNB), laparoscopic cholecystectomy (LC), laparoscopic inguinal hernia repair (LIH), and open inguinal hernia repair (IH). We found that opioids were over-prescribed. We formulated guidelines for opioid prescribing that would halve the number of pills prescribed and also satisfy 80% of patients' opioid requirements. We discussed our findings and opioid-prescribing guidelines with surgeons at our institution. We recommended that surgeons encourage patients to use a nonsteroidal anti-inflammatory drug (NSAID) and acetaminophen before using opioids. We then evaluated opioid prescriptions and use in 246 subsequent patients undergoing these same operations. The mean number of opioid pills prescribed for each operation markedly decreased: PM 19.8 versus 5.1; PM SLNB 23.7 versus 9.6; LC 35.2 versus 19.4; LIH 33.8 versus 19.3, and IH 33.2 versus 18.3; all P < 0.0003. The total number of pills prescribed decreased by 53% when compared with the number that would have been prescribed before the educational intervention. Only 1 patient (0.4%) required a refill opioid prescription. Eighty-five percent of patients used either a NSAID or acetaminophen. By defining postoperative opioid requirements through patient surveys and disseminating operation-specific guidelines for opioid prescribing to surgeons, we were able to decrease the number of opioids initially prescribed by more than half. Decreased initial opioid prescriptions did not result in increased opioid refill prescriptions.


A critical overview of the current myofascial pain literature - March 2016.
The worldwide interest in myofascial pain syndrome (MPS) and trigger points (TrPs) is reflected in the increasing number of publications. In this overview of the literature, we included 26 studies, case reports and review articles by authors from 18 different countries. Several research groups are exploring the characteristic of TrPs such as Chen and colleagues, who continued their work on the quantification of the taut bands. Meng and colleagues studied the relationships between TrPs and central sensitization, while Yu and colleagues examined the electrophysiological characteristics that occur as a result of active TrPs. Several researchers used objective measurements to determine clinical outcomes, such as Koppenhaver and colleagues who measured objective changes in the function and nociceptive sensitivity of lumbar multifidus muscle subjects with low back pain. Turo and colleagues quantified muscle tissue changes after dry needling in chronic myofascial pain using elastography. Multiple studies explored various treatment options for TrPs, such as dry needling, injections with lidocaine or granisetron, traditional Thai massage, self-myofascial release, kinesiotaping, and monochromatic infrared photo energy, among others.


Release of vasoactive intestinal peptide in mouse cerebral cortex: evidence for a role of arachidonic acid metabolites.
In rodent cerebral cortex, vasoactive intestinal peptide (VIP) is contained in a homogeneous population of radially oriented bipolar interneurons. We have observed that 4-aminopyridine (4-AP), a K+-channel blocker, promotes a concentration- and Ca2+-dependent release of VIP from mouse cerebral cortical slices, with a significant effect already observed at 50 microM. Over 70% of VIP release elicited by 4-AP is blocked by 2 microM tetrodotoxin (TTX). Mepacrine, an inhibitor of phospholipase A2 (PLA2) activity and hence of arachidonic acid (AA) formation from membrane phospholipids, markedly inhibits (IC50 of approximately 15 microM) the release of VIP evoked by 4-AP. The inhibitory effect of mepacrine is not additive to that of TTX, thus indicating an involvement of PLA2 activation in the TTX-sensitive component of the 4-AP-evoked release. As a corollary, melittin (0.1-10 micrograms/ml), a PLA2 activator, promotes VIP release. Inhibition of AA metabolites of the lipoxygenase pathway by nordihydroguaiaretic acid, 5,8,11,14-eicosatetranoic acid, and caffeic acid results in a concentration-dependent inhibition of VIP release evoked by 4-AP. This set of observations indicates for the first time that the formation of AA metabolites of the lipoxygenase pathway plays a role in the release of a peptide in the mammalian CNS. Furthermore, these observations together with the previously reported potentiation by prostaglandins of the increase in cyclic AMP elicited by VIP in mouse cerebral cortex (Schaad et al., 1987) indicate that AA metabolites may act at both the presynaptic (lipoxygenase metabolites) and the postsynaptic (cyclooxygenase metabolites) levels to increase the "throughput" or "strength" of VIP-containing circuits in the rodent neocortex.


Monitoring and Analysis Solid Formulation Dissolution Phenomenon with Image Recognition Technologies.
The dissolution test has become the most important quality index in the research and development of solid formulation, especially the evaluation of drug bioequivalence. However, it had low operability, was tedious, and was always overlooked. The previously related studies required a fixed tablet and analysed the recorded video by disso GUARO PRO and Microsoft Paint™. Therefore, we have developed a novel image recognition system to automatically track the moving tablet and analyse the volume change at the same time. Image recognition technology is often used to monitor the dissolution process. The camera system with visible light and infrared camera functions was placed on the dissolution tester. The system collects the plate image for binary processing and then records and calculates its pixel area, which can automatically record the volume change of the tablet in the dissolution test, no matter disintegration or corrosion.


Building the foundations of an informatics agenda for global health - 2011 workshop report.
Strengthening the capacity of public health systems to protect and promote the health of the global population continues to be essential in an increasingly connected world. Informatics practices and principles can play an important role for improving global health response capacity. A critical step is to develop an informatics agenda for global health so that efforts can be prioritized and important global health issues addressed. With the aim of building a foundation for this agenda, the authors developed a workshop to examine the evidence in this domain, recognize the gaps, and document evidence-based recommendations. On 21 August 2011, at the 2011 Public Health Informatics Conference in Atlanta, GA, USA, a four-hour interactive workshop was conducted with 85 participants from 15 countries representing governmental organizations, private sector companies, academia, and non-governmental organizations. The workshop discussion followed an agenda of a plenary session - planning and agenda setting - and four tracks: Policy and governance; knowledge management, collaborative networks and global partnerships; capacity building; and globally reusable resources: metrics, tools, processes, templates, and digital assets. Track discussions examined the evidence base and the participants' experience to gather information about the current status, compelling and potential benefits, challenges, barriers, and gaps for global health informatics as well as document opportunities and recommendations. This report provides a summary of the discussions and key recommendations as a first step towards building an informatics agenda for global health. Attention to the identified topics and issues is expected to lead to measurable improvements in health equity, health outcomes, and impacts on population health. We propose the workshop report be used as a foundation for the development of the full agenda and a detailed roadmap for global health informatics activities based on further contribution from key stakeholders. The global health informatics agenda and roadmap can provide guidance to countries for developing and enhancing their individual and regional agendas.


Diagnostic and prognostic value of microRNA-628 for cancers.
Background: Many studies manifested miRNA-628 (miR-628) was deregulated in various cancers, indicating that miR-628 might serve as a novel biomarker of cancer diagnosis and prognosis, but it's role was still uncertain. This study aimed to evaluate the value of miR-628 in various cancers for diagnosis and prognosis, as well as its predictive power in combination biomarkers. Materials and Methods: A literature search was performed using Medline (via PubMed), Embase, Web of Science databases, and Ovid platform up to November 2017. Meta-analysis was performed to provide summative outcomes. Quality assessment of each included study was performed. Results: Twelve articles with 20 studies were included in our meta-analysis, including 8 articles with 15 studies for diagnostic meta-analysis and 4 articles with 5 studies for prognostic meta-analysis. For the diagnostic meta-analysis of miR-628 alone, the overall pooled results for sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and the area under the summary receiver operating characteristic (SROC) curve (AUC) were 0.81 (95% CI: 0.62-0.91), 0.72 (95% CI: 0.48-0.88), 2.90 (95% CI: 1.50-5.40), 0.27 (95% CI: 0.14-0.50), 11.0 (95% CI: 4.00-25.00), and 0.84 (95% CI: 0.80-0.87), respectively. For the diagnostic meta-analysis of miR-628-related combination biomarkers, the above six parameters were 0.89 (95% CI: 0.84-0.92), 0.93 (95% CI: 0.82-0.97), 12.30 (95% CI: 4.70-32.50), 0.12 (95% CI: 0.08-0.19), and 100.00 (95% CI: 28.00-354.00), 0.93 (95% CI: 0.90-0.95), respectively. For the prognostic meta-analysis, patients with lower miR-628 had significant shorter overall survival than high expression of miR-628 (HR = 1.553, 95% CI: 1.041-2.318, z = 2.16, P = 0.031). Conclusions: This study confirms that miR-628 may be a promising biomarker for cancer diagnosis and prognosis. Expertly, microRNAs combination biomarkers could be a new alternative for clinical application.


A high-content small molecule screen identifies sensitivity of glioblastoma stem cells to inhibition of polo-like kinase 1.
Glioblastoma multiforme (GBM) is the most common primary brain cancer in adults and there are few effective treatments. GBMs contain cells with molecular and cellular characteristics of neural stem cells that drive tumour growth. Here we compare responses of human glioblastoma-derived neural stem (GNS) cells and genetically normal neural stem (NS) cells to a panel of 160 small molecule kinase inhibitors. We used live-cell imaging and high content image analysis tools and identified JNJ-10198409 (J101) as an agent that induces mitotic arrest at prometaphase in GNS cells but not NS cells. Antibody microarrays and kinase profiling suggested that J101 responses are triggered by suppression of the active phosphorylated form of polo-like kinase 1 (Plk1) (phospho T210), with resultant spindle defects and arrest at prometaphase. We found that potent and specific Plk1 inhibitors already in clinical development (BI 2536, BI 6727 and GSK 461364) phenocopied J101 and were selective against GNS cells. Using a porcine brain endothelial cell blood-brain barrier model we also observed that these compounds exhibited greater blood-brain barrier permeability in vitro than J101. Our analysis of mouse mutant NS cells (INK4a/ARF(-/-), or p53(-/-)), as well as the acute genetic deletion of p53 from a conditional p53 floxed NS cell line, suggests that the sensitivity of GNS cells to BI 2536 or J101 may be explained by the lack of a p53-mediated compensatory pathway. Together these data indicate that GBM stem cells are acutely susceptible to proliferative disruption by Plk1 inhibitors and that such agents may have immediate therapeutic value.


Habitual use of GPS negatively impacts spatial memory during self-guided navigation.
Global Positioning System (GPS) navigation devices and applications have become ubiquitous over the last decade. However, it is unclear whether using GPS affects our own internal navigation system, or spatial memory, which critically relies on the hippocampus. We assessed the lifetime GPS experience of 50 regular drivers as well as various facets of spatial memory, including spatial memory strategy use, cognitive mapping, and landmark encoding using virtual navigation tasks. We first present cross-sectional results that show that people with greater lifetime GPS experience have worse spatial memory during self-guided navigation, i.e. when they are required to navigate without GPS. In a follow-up session, 13 participants were retested three years after initial testing. Although the longitudinal sample was small, we observed an important effect of GPS use over time, whereby greater GPS use since initial testing was associated with a steeper decline in hippocampal-dependent spatial memory. Importantly, we found that those who used GPS more did not do so because they felt they had a poor sense of direction, suggesting that extensive GPS use led to a decline in spatial memory rather than the other way around. These findings are significant in the context of society's increasing reliance on GPS.


Incidental statistical summary representation over time.
Information taken in by the human visual system allows individuals to form statistical representations of sets of items. One's knowledge of natural categories includes statistical information, such as average size of category members and the upper and lower boundaries of the set. Previous research suggests that when subjects attend to a particular dimension of a set of items presented over an extended duration, they quickly learn about the central tendency of the set. However, it is unclear whether such learning can occur incidentally, when subjects are not attending to the relevant dimension of the set. The present study explored whether subjects could reproduce global statistical properties of a set presented over an extended duration when oriented to task-irrelevant properties of the set. Subjects were tested for their memory of its mean, its smallest and largest exemplars, the direction of its skew, and the relative distribution of the items. Subjects were able to accurately recall the average size circle, as well as the upper and lower boundaries of a set of 4,200 circles displayed over an extended period. This suggests that even without intending to do so, they were encoding and updating a statistical summary representation of a task-irrelevant attribute of the circles over time. Such incidental encoding of statistical properties of sets is thus a plausible mechanism for establishing a representation of typicality in category membership.


Use and satisfaction with key functions of a common commercial electronic health record: a survey of primary care providers.
Despite considerable financial incentives for adoption, there is little evidence available about providers' use and satisfaction with key functions of electronic health records (EHRs) that meet "meaningful use" criteria. We surveyed primary care providers (PCPs) in 11 general internal medicine and family medicine practices affiliated with 3 health systems in Texas about their use and satisfaction with performing common tasks (documentation, medication prescribing, preventive services, problem list) in the Epic EHR, a common commercial system. Most practices had greater than 5 years of experience with the Epic EHR. We used multivariate logistic regression to model predictors of being a structured documenter, defined as using electronic templates or prepopulated dot phrases to document at least two of the three note sections (history, physical, assessment and plan). 146 PCPs responded (70%). The majority used free text to document the history (51%) and assessment and plan (54%) and electronic templates to document the physical exam (57%). Half of PCPs were structured documenters (55%) with family medicine specialty (adjusted OR 3.3, 95% CI, 1.4-7.8) and years since graduation (nonlinear relationship with youngest and oldest having lowest probabilities) being significant predictors. Nearly half (43%) reported spending at least one extra hour beyond each scheduled half-day clinic completing EHR documentation. Three-quarters were satisfied with documenting completion of pneumococcal vaccinations and half were satisfied with documenting cancer screening (57% for breast, 45% for colorectal, and 46% for cervical). Fewer were satisfied with reminders for overdue pneumococcal vaccination (48%) and cancer screening (38% for breast, 37% for colorectal, and 31% for cervical). While most believed the problem list was helpful (70%) and kept an up-to-date list for their patients (68%), half thought they were unreliable and inaccurate (51%). Dissatisfaction with and suboptimal use of key functions of the EHR may mitigate the potential for EHR use to improve preventive health and chronic disease management. Future work should optimize use of key functions and improve providers' time efficiency.


The effect of amino acids on the metabolic fate of 15NH4Cl in isolated sheep hepatocytes.
Ruminants characteristically absorb a large proportion of dietary nitrogen across the portal-drained viscera as ammonia nitrogen which is detoxified by conversion to urea in the liver. In theory, ammonia can supply both nitrogen atoms of the urea molecule via mitochondrial (carbamoyl phosphate) and cytoplasmic (aspartate) precursor pathways of the ornithine cycle but the effect of amino acids on the flux of nitrogen from ammonia to each of the two urea nitrogen atoms has not been determined. We report a study designed to determine the distribution of [15N] ammonia between [15N1]urea and [15N2]urea in sheep hepatocytes in response to ammonia concentrations (0.33, 0.67 and 1.00 mM) in the presence or absence of amino acids. In the absence of amino acids, the enrichment of [15N2]urea rose more rapidly during incubations than [15N1]urea and attained enrichments of 66-88% within 5 min of incubation. At the end of 2.5 h of incubation, [15N2]urea represented 60% and 90% of the total urea molecules at low and high ammonia concentrations, respectively. The enrichments of glutamate and aspartate were similar to [15N1]urea in the cells at the end of the incubations, even in the presence of unlabelled amino acids, supporting the concept of mitochondrial ammonia being in equilibrium with cytosolic aspartate formation. In the presence of amino acids basal urea synthesis increased but ammonia uptake and 15NH4Cl conversion to urea was less than in the absence of amino acids. The rate of formation of [15N1]urea was greater in incubations containing amino acids but when ammonia concentration in the media was raised only [15N2]urea flux increased with no change in either [15N1]urea or the unlabelled species. Measurement of media amino acid concentrations after 2.5 h of incubation in the presence of amino acids revealed that arginine, glutamine, glycine and alanine were removed while there was net formation of aspartate, threonine, serine, glutamate, and the branched chain amino acids. However, less than 12% of the 15N transfer appeared in free amino acids. The increases in basal and unlabelled urea synthesis in the presence of amino acids could be numerically accounted as the sum of arginine and glutamine removal from incubations. It is concluded that in sheep hepatocytes 15NH4Cl removal leads to quantitative formation of [15N2]urea, even in the presence of a physiological mixture of amino acids. The increase in the formation of the [15N1]urea in the presence of amino acids can be explained by the preferential utilisation of the amide nitrogen of glutamine for urea synthesis.


Transport of N1-methylnicotinamide across brush border membrane vesicles from rabbit kidney.
A preparation of isolated brush border membranes from the rabbit renal cortex was used to examine the characteristics of N1-methylnicotinamide (NMN) transport in the kidney. Transport was independent of the presence of Na+ under Na+ equilibrium conditions. However, outwardly directed Na+ gradients stimulated NMN uptake, whereas inwardly directed Na+ gradients inhibited NMN uptake. Transport appeared to involve two parallel processes: one saturable with a Jmax of 5 nmol X mg protein-1 X min-1 and an apparent Kt of 0.6 mM, and a second that behaved like passive diffusion. Countertransport of NMN was observed when vesicles were preloaded with either NMN or another organic cation, tetraethylammonium (TEA). TEA and several structural analogues, as well as a wide variety of other organic cations and bases, were effective inhibitors of NMN uptake, though nicotinamide and p-aminohippuric acid did not interact with the uptake process. Outwardly directed proton gradients (pH 6.0 in, 7.6 out) stimulated transport, suggesting that NMN uptake may involve a countertransport of H+. The electrical potential difference across the vesicle membrane was manipulated using gradients of the permeant organic ion thiocyanate (SCN-); an outwardly directed gradient of SCN- (i.e., a depolarizing condition) stimulated uptake and produced a transient accumulation of NMN above that noted at equilibrium, whereas an inwardly directed SCN- gradient inhibited uptake of NMN. The data can be explained by postulating the presence of an electrogenic NMN+-H+ antiporter in the rabbit luminal membrane that could play a role in organic cation secretion.


Weak interaction of spectrin with phosphatidylcholine-phosphatidylserine multilayers: a 2H and 31P NMR study.
Spectrin from human erythrocytes binds to bilayer dispersions of both DMPC and DMPS:DMPC (1:1, w/w). However, no effect of bound spectrin on the conformation of the lipid head groups, as measured from the deuterium quadrupolar splittings of DMPC or DMPS specifically deuterated in the polar head groups, was detected in 1:1 mixtures of the two lipids containing either deuterated DMPC or DMPS. Neither the phase transition of the DMPS:DMPC mixtures, nor the spin-lattice relaxation time (T1) of the deuterated DMPS head group, was affected by spectrin. These results argue against any strong interaction of spectrin with phosphatidylserine and rule out the possibility that spectrin is responsible for the maintenance of PS in the inner monolayer of the erythrocyte membrane during the whole life-span of this cell.


[The effect of antibiotics on the development of enterotoxemia-- experimental study].
When testing antibiotics in rabbits Dalacin was shown to be the most effective in inducing enterotoxaemia in rabbits. Autopsy disclosed signs of enterotoxaemia even in some animals without previous symptoms. Histopathological lesions were found even in intestines macroscopically normal. Experiments proved that after application of tested antibiotics spread through intestinal mucosa and produced a loss of microbial balance. Rabbit is to be considered suitable for modelling antibiotic-induced enterotoxaemia.


The design of a contextualized responsive evaluation framework for fishery management in Benin.
The main question addressed by this article is how to adapt the responsive evaluation (RE) approach to an intervention context characterized by repetition of ineffective interventions, ambiguous intervention action theories among stakeholders, and high complexity. The context is Grand-Popo, a fishing municipality located on Benin's southwest Atlantic coast. The fishery management interventionists and the fishing communities in the municipality all espoused concern for the sustainable improvement of fishing actors' livelihood conditions, but differed about the reasons for this livelihood impairment, and about what should be done, when, where, and by whom. Given this ambiguity, we identified RE as a promising action research approach to facilitate dialogue and mutual learning, and consequently to improve stakeholders' ability to resolve problems. However, this approach seems to have some shortcomings in the Grand-Popo context, regarding the repetitive ineffectiveness of interventions, high complexity, and uncertainty. Therefore, based on our empirical study, we add three dimensions to the existing RE framework: historical analysis to deal with routine interventions, exploration and discussion of incongruities of action theories to trigger double-loop learning, and system analysis to deal with complexity and uncertainty. This article does not intend to address the implications or impact of this adapted RE framework. Instead, we suggest some criteria and indicators for evaluating whether the proposed amended RE approach has assisted in resolving the fishery problems in Grand-Popo after the approach has been applied.


The validity of a rheumatoid arthritis medical records-based index of severity compared with the DAS28.
Outcome measures play an extremely important role in clinical trials and observational research. Outcome measures for rheumatoid arthritis cover a whole array of domains, ranging from measures describing the inflammatory process to measures describing the ultimate consequences of long-term disease, such as joint damage, physical function and quality of life. There is a scientific need to be able to quantify what is called the 'severity of rheumatoid arthritis', so that patients with rheumatoid arthritis can be clustered according to their propensity to develop an unfavourable outcome. It is a challenge to find an appropriate measure for severity. One attempt has been the development of the Rheumatoid Arthritis Medical Record-Based Index of Severity. This commentary elaborates on how such a measure of severity should be validated to determine whether it is appropriate for practical use.


Effectiveness of the HomeSafe Pilot Program in reducing injury rates among residential construction workers, 1994-1998.
The construction industry typically has one of the highest fatal and non-fatal injury rates compared with other industries. Residential construction workers are at particular risk of injury (work is in remote sites with small crews, there are often many subcontractors, and they have limited access to safety programs). Difficulty accessing information specific to this group has made research more challenging, therefore, there are few studies. This study evaluated the effectiveness of the HomeSafe Pilot Program, a safety education and training program designed to reduce injuries among residential construction workers. Researchers evaluated whether overall and severe injury incidence rates declined during the intervention period. Data were analyzed using incidence rates and Poisson regression to control for the effect of antecedent secular trend. Injury incidence rates declined significantly following HomeSafe; however, this effect was not statistically significant once temporal variation was controlled. The decline in injury rates following HomeSafe cannot be attributed solely to HomeSafe, however, programmatic and methodologic limitations contributed to the inconclusive results. Further research into the hazards faced by residential construction workers is needed. Am. J. Ind. Med. 45:210-217, 2004.


Spatially adaptive wavelet thresholding with context modeling for image denoising.
The method of wavelet thresholding for removing noise, or denoising, has been researched extensively due to its effectiveness and simplicity. Much of the literature has focused on developing the best uniform threshold or best basis selection. However, not much has been done to make the threshold values adaptive to the spatially changing statistics of images. Such adaptivity can improve the wavelet thresholding performance because it allows additional local information of the image (such as the identification of smooth or edge regions) to be incorporated into the algorithm. This work proposes a spatially adaptive wavelet thresholding method based on context modeling, a common technique used in image compression to adapt the coder to changing image characteristics. Each wavelet coefficient is modeled as a random variable of a generalized Gaussian distribution with an unknown parameter. Context modeling is used to estimate the parameter for each coefficient, which is then used to adapt the thresholding strategy. This spatially adaptive thresholding is extended to the overcomplete wavelet expansion, which yields better results than the orthogonal transform. Experimental results show that spatially adaptive wavelet thresholding yields significantly superior image quality and lower MSE than the best uniform thresholding with the original image assumed known.


Association of Midlife Cardiovascular Risk Profiles With Cerebral Perfusion at Older Ages.
Poor cardiovascular health is an established risk factor for dementia, but little is known about its association with brain physiology in older adults. To examine the association of cardiovascular risk factors, measured repeatedly during a 20-year period, with cerebral perfusion at older ages. In this longitudinal cohort study, individuals were selected from the Whitehall II Imaging Substudy. Participants were included if they had no clinical diagnosis of dementia, had no gross brain structural abnormalities on magnetic resonance imaging scans, and had received pseudocontinuous arterial spin labeling magnetic resonance imaging. Cardiovascular risk was measured at 5-year intervals across 5 phases from September 1991 to October 2013. Arterial spin labeling scans were acquired between April 2014 and December 2014. Data analysis was performed from June 2016 to September 2018. Framingham Risk Score (FRS) for cardiovascular disease, comprising age, sex, high-density lipoprotein cholesterol level, total cholesterol level, systolic blood pressure, use of antihypertensive medications, cigarette smoking, and diabetes, was assessed at 5 visits. Cerebral blood flow (CBF; in milliliters per 100 g of tissue per minute) was quantified with pseudocontinuous arterial spin labeling magnetic resonance imaging. Of 116 adult participants, 99 (85.3%) were men. At the first examination, mean (SD) age was 47.1 (5.0) years; at the last examination, mean (SD) age was 67.4 (4.9) years. Mean (SD) age at MRI scan was 69.3 (5.0) years. Log-FRS increased with time (B = 0.058; 95% CI, 0.044 to 0.072; P < .001). Higher cumulative FRS over the 20-year period (measured as the integral of the rate of change of log-FRS) was associated with lower gray matter CBF (B = -0.513; 95% CI -0.802 to -0.224; P < .001) after adjustment for age, sex, education, socioeconomic status, cognitive status, arterial transit time, use of statins, and weekly alcohol consumption. Voxelwise analyses revealed that this association was significant in 39.6% of gray matter regions, including the posterior cingulate, precuneus, lateral parietal cortex, occipital cortex, hippocampi, and parahippocampal gyrus. The strength of the association of higher log-FRS with lower CBF decreased progressively from the first examination (R2 = 0.253; B = -10.816; 99% CI -18.375 to -3.257; P < .001) to the last (R2 = 0.188; B = -7.139; 99% CI -14.861 to 0.582; P = .02), such that the most recent FRS measurement at mean (SD) age 67.4 (4.9) years was not significantly associated with CBF with a Bonferroni-corrected P < .01 . Cardiovascular risk in midlife was significantly associated with lower gray matter perfusion at older ages, but this association was not significant for cardiovascular risk in later life. This finding could inform the timing of cardiovascular interventions so as to be optimally effective.


Genetics of protein I of Neisseria gonorrhoeae: construction of hybrid porins.
Protein I (PI), the major outer membrane protein of Neisseria gonorrhoeae, is a porin and occurs in two major immunochemical classes, A and B. By using shuttle mutagenesis to insert a selectable marker close to the PI structural gene, evidence was obtained from transformation experiments to demonstrate that the PI structural gene is equivalent to the defined locus nmp and that the genes for PI class A and PI class B are alleles of the same locus. The PI class B gene of strain MS11 was cloned and sequenced, and comparison of this sequence with the gene sequence of PI class A of FA19 revealed a number of regions of significant divergence. By selection for the closely linked marker in transformations between the two strains, a series of strains with a hybrid PI was obtained. Analysis of these strains with monoclonal antibodies and oligonucleotides specific to PI class A or PI class B elucidated the nature and location of some of the surface-exposed epitopes, a thorough characterization of which is a prerequisite for understanding the role of PI in gonococcal pathogenesis and its possible use as a component of a vaccine.


Identification of grapevine clones via high-throughput amplicon sequencing: a proof-of-concept study.
Wine cultivars are available to growers in multiple clonal selections with agronomic and enological differences. Phenotypic differences between clones originated from somatic mutations that accrued over thousands of asexual propagation cycles. Genetic diversity between grape cultivars remains unexplored, and tools to discriminate unequivocally clones have been lacking. This study aimed to uncover genetic variations among a group of clonal selections of 4 important Vitis vinifera cultivars: Cabernet sauvignon, Sauvignon blanc, Chardonnay, and Merlot, and use this information to develop genetic markers to discriminate the clones of these cultivars. We sequenced with short-read sequencing technology the genomes of 18 clones, including biological replicates for a total of 46 genomes. Sequences were aligned to their respective cultivar's reference genome for variant calling. We used reference genomes of Cabernet sauvignon, Chardonnay, and Merlot and developed a de novo genome assembly of Sauvignon blanc using long-read sequencing. On average, 4 million variants were detected for each clone, with 74.2% being single nucleotide variants and 25.8% being small insertions or deletions (InDel). The frequency of these variants was consistent across all clones. From these variants, we validated 46 clonal markers using high-throughput amplicon sequencing for 77.7% of the evaluated clones, most of them small InDel. These results represent an advance in grapevine genotyping strategies and will benefit the viticulture industry for the characterization and identification of the plant material.


Molecular cloning and characterization of p56dok-2 defines a new family of RasGAP-binding proteins.
Chronic myelogenous leukemia (CML) is a disease characterized by the presence of p210(bcr-abl), a chimeric protein with tyrosine kinase activity. Substrates for p210(bcr-abl) are likely to be involved in the pathogenesis of CML. Here we describe the purification, cDNA cloning, and characterization of a 56-kDa tyrosine phosphorylated protein, p56(dok-2) (Dok-2), from p210(bcr-abl) expressing cells. The human dok-2 cDNA encodes a 412-amino acid protein with a predicted N-terminal pleckstrin homology domain as well as several other features of a signaling molecule, including 13 potential tyrosine phosphorylation sites, six PXXP motifs, and the ability to bind to p120(RasGAP). Dok-2 was shown to be 35% identical to p62(dok-1), a recently identified RasGAP binding protein from CML cells, and analysis of the expressed sequence tag data base revealed the presence of at least four additional proteins containing a Dok homology sequence motif. Dok mRNAs were primarily expressed in tissues of hematopoietic origin. These findings strongly suggest that a family of Dok-related proteins exists that bind to RasGAP and may mediate the effects of p210(bcr-abl) in CML.


Niclosamide prevents the formation of large ubiquitin-containing aggregates caused by proteasome inhibition.
Protein aggregation is a hallmark of many neurodegenerative diseases and has been linked to the failure to degrade misfolded and damaged proteins. In the cell, aberrant proteins are degraded by the ubiquitin proteasome system that mainly targets short-lived proteins, or by the lysosomes that mostly clear long-lived and poorly soluble proteins. Both systems are interconnected and, in some instances, autophagy can redirect proteasome substrates to the lysosomes. To better understand the interplay between these two systems, we established a neuroblastoma cell population stably expressing the GFP-ubiquitin fusion protein. We show that inhibition of the proteasome leads to the formation of large ubiquitin-containing inclusions accompanied by lower solubility of the ubiquitin conjugates. Strikingly, the formation of the ubiquitin-containing aggregates does not require ectopic expression of disease-specific proteins. Moreover, formation of these focused inclusions caused by proteasome inhibition requires the lysine 63 (K63) of ubiquitin. We then assessed selected compounds that stimulate autophagy and found that the antihelmintic chemical niclosamide prevents large aggregate formation induced by proteasome inhibition, while the prototypical mTORC1 inhibitor rapamycin had no apparent effect. Niclosamide also precludes the accumulation of poly-ubiquitinated proteins and of p62 upon proteasome inhibition. Moreover, niclosamide induces a change in lysosome distribution in the cell that, in the absence of proteasome activity, may favor the uptake into lysosomes of ubiquitinated proteins before they form large aggregates. Our results indicate that proteasome inhibition provokes the formation of large ubiquitin containing aggregates in tissue culture cells, even in the absence of disease specific proteins. Furthermore our study suggests that the autophagy-inducing compound niclosamide may promote the selective clearance of ubiquitinated proteins in the absence of proteasome activity.


The quality and effectiveness of interventions that target multiple risk factors among young people: a systematic review.
To identify evaluations of interventions that target multiple risk factors in high-risk young people, describe their characteristics, critique their methodological quality and summarise their effectiveness. A search of the literature published between 2009 and 2014 identified 13 evaluations of interventions that targeted multiple risk factors, compared to 95 evaluations that targeted single risk factors. The methodological adequacy of the 13 evaluation studies was analysed using the Quality Assessment Tool for Quantitative Studies and information regarding characteristics and intervention effectiveness was extracted and summarised. There were very few outcome evaluation studies of interventions that targeted multiple risk factors, relative to single risk factors, among high-risk young people. Of the identified studies, half were methodologically weak. Interventions delivered in community settings targeted a greater number of risk factors, while those delivered in a school or health setting reported a higher proportion of statistically significant outcomes. No economic analyses were conducted. Conclusions and Implications for Public Health: More methodologically rigorous evaluations of interventions targeting multiple risk factors among high-risk young people are required, especially for those delivered in community settings. Four key areas for improvement are: i) more precisely defining the risk factors experienced by high-risk young people; ii) achieving greater consistency across interventions; iii) standardising outcome measures; and iv) conducting economic analyses.


Paternal incarceration and support for children in fragile families.
High U.S. incarceration rates have motivated recent research on the negative effects of imprisonment on later employment, earnings, and family relationships. Because most men in jail and prison are fathers, a large number of children may be placed at considerable risk by policies of incarceration. This article examines one dimension of the economic risk faced by children of incarcerated fathers: the reduction in the financial support that they receive. We use a population-based sample of urban children to examine the effects of incarceration on this support. Both cross-sectional and longitudinal regressions indicate that formerly incarcerated men are less likely to contribute to their families, and those who do contribute provide significantly less. The negative effects of incarceration on fathers' financial support are due not only to the low earnings of formerly incarcerated men but also to their increased likelihood to live apart from their children. Men contribute far less through child support (formal or informal) than they do when they share their earnings within their household, suggesting that the destabilizing effects of incarceration on family relationships place children at significant economic disadvantage.


Biosynthetic origins of menisporopsin A.
Menisporopsin A, produced by Menisporopsis theobromae, shows antimalarial, antimycobacterial, and cytotoxic activities. Here, we report the first (13)C incorporations at individual carbons of menisporopsin A using sodium [1-(13)C] and [2-(13)C] acetate. This result indicates that each of the subunits of the pentalactone menisporopsin A is assembled by a polyketide synthase.


Smell identification function as a severity and progression marker in Alzheimer's disease.
Olfactory dysfunction, impaired smell identification in particular, is known as a diagnostic and a marker of conversion in Alzheimer's disease (AD). We aimed to evaluate the associations of olfactory identification impairments with cognition, illness severity, and progression in AD patients. Fifty-seven outpatients with late onset mild to moderate AD and 24 elderly non-demented controls (NDC) were assessed, at baseline and after three months, for Mini-Mental State Examination (MMSE), University of Pennsylvania Smell Identification Test (UPSIT), and Bristol Activities of Daily Living and Neuropsychiatry Inventory. AD participants were classified as Rapid Cognitive Decliners (RCD) defined on a priori with a loss of ≥2 points in MMSE within the previous six months. AD participants had lower olfactory scores than NDC. RCD had lower olfaction scores compared with Non-Rapid Cognitive Decliners (NRCD). Although the baseline UPSIT scores were associated with baseline MMSE scores, it did not interact significantly with change in MMSE over the follow-up period. Using a median split for olfactory scores, the AD participants were classified as Rapid Olfactory Progressors (ROP) (UPSIT ≤ 15) and Slow Olfactory Progressors correlating significantly with RCD/NRCD groups. The ROP group with higher olfactory impairment indicated more symptomatic illness or severity, i.e. lower cognition, higher functional dependence, and presence of behavioral symptoms. Our study supports association of smell identification function with cognition and its utility as an adjunct clinical measure to assess severity in AD. Further work, including larger longitudinal studies, is needed to explore its value in predicting AD progression.


A smartphone-based chip-scale microscope using ambient illumination.
Portable chip-scale microscopy devices can potentially address various imaging needs in mobile healthcare and environmental monitoring. Here, we demonstrate the adaptation of a smartphone's camera to function as a compact lensless microscope. Unlike other chip-scale microscopy schemes, this method uses ambient illumination as its light source and does not require the incorporation of a dedicated light source. The method is based on the shadow imaging technique where the sample is placed on the surface of the image sensor, which captures direct shadow images under illumination. To improve the image resolution beyond the pixel size, we perform pixel super-resolution reconstruction with multiple images at different angles of illumination, which are captured while the user is manually tilting the device around any ambient light source, such as the sun or a lamp. The lensless imaging scheme allows for sub-micron resolution imaging over an ultra-wide field-of-view (FOV). Image acquisition and reconstruction are performed on the device using a custom-built Android application, constructing a stand-alone imaging device for field applications. We discuss the construction of the device using a commercial smartphone and demonstrate the imaging capabilities of our system.


Public support for street-scale urban design practices and policies to increase physical activity.
Street-scale urban design policies are recommended to increase physical activity in communities. Our purpose was to examine U.S. public support for local street-scale urban design features and policies. Analysis is based on a cross-sectional national sample of adults (n = 4682) participating in the 2006 HealthStyles mail survey. About 57% of adults rated local street-scale urban design as highly important in determining the amount of physical activity they obtain. Adjusted odds of rating neighborhood features as having high importance were higher in people aged ≥65 years versus those <65 and minority racial/ethnic groups versus non-Hispanic whites. Two-thirds of adults were willing to take civic action to support local street-scale urban design policy. Adjusted odds of being willing to take any action versus none was higher in non-Hispanic blacks and Hispanics versus non-Hispanic whites, was higher in those with household incomes ≥$60,000 versus ≤$15,000 per year, and increased as education and perceived importance of neighborhood features increased. There are high levels of public support for local street-scale urban design policies; however, demographic differences exist in the level of support. These differences are important considerations for policymakers and for those designing community programs targeting street-scale urban design features and policies.


Development stage-specific expression of fibroin in the silk worm Bombyx mori is regulated translationally.
The contents of fibroin H RNA as a function of development have been quantitated in the posterior silk glands of Bombyx mori larvae on different days of 4th and 5th instars. The fibroin RNA levels increased during the feeding stages of larvae and the RNA got completely degraded during the interim moult. The patterns of accumulation of fibroin RNA were similar in both the instars. Although there was considerable increase in the fibroin RNA content during the 5th larval instar, the relative abundance of fibroin RNA in the total RNA was fairly constant during the 4th and 5th instars. The increased content of fibroin RNA in 5th instar was the consequence of an overall increase in transcription accompanying the development progress, rather than specific increase only in fibroin transcription. The contents of fibroin protein in the 4th and 5th instars of development have also been quantitated making use of a sensitive radioimmune assay with a purified, antifibroin antibody. There were substantial differences between 4th and 5th instars in the absolute fibroin contents as well as the relative proportion of fibroin in the total proteins. These results implied that although the fibroin gene was transcribed at the same efficiency during the 4th and 5th instars, the translational efficiency was much lower during the 4th instar. The extent of polyadenylation of fibroin RNA was similar in both instars. However, there was a two-fold increase in the polysome association of fibroin RNA in the 5th instar. Over and above this, there was substantial increase during the 5th instar in the contents of those tRNAs. (e.g. Gly, Ala and Ser) which are abundantly represented in fibroin and therefore directly related to the expression of fibroin. The increased polysome association of fibroin mRNA and the adequate supply of cognate tRNAs in the 5th instar, together contributes to the translational regulation of fibroin in a developmental stage-specific manner. Based on these observations, we propose that translational regulation plays a major role in the development stage-specific synthesis of fibroin in Bombyx mori.


Duck populations as indicators of landscape condition in the Prairie Pothole Region.
The Prairie Pothole Region of the northern Great Plains is an important region for waterfowl production because of the abundance of shallow wetlands. The ecological significance of the region and impacts from intensive agriculture prompted the U.S. Environmental Protection Agency to select it as one of the first areas for developing and evaluating ecological indicators of wetland condition. We examined hypothesized relations between indicators of landscape and wetland conditions and waterfowl abundance on 45 40 km2 study sites in North Dakota for 1995-1996. Landscape condition was defined a priori as the ratio of cropland area to total upland area surrounding wetlands. Measures of waterfowl abundance included estimated numbers of breeding pairs (by species and total numbers) and gamma, a species-specific correction factor which effectively adjusts breeding pair estimates for annual or area-related differences in pond size. Landscape indicators and waterfowl measures varied among regions. Results indicated that most areas in the Coteau region are of much higher quality for ducks than those in the Drift Plain, and areas in the Red River Valley are of the poorest quality for ducks. Regression models demonstrated the impact of agricultural development on breeding duck populations in the Prairie Pothole Region. The most consistent landscape indicators of waterfowl abundance were percent of cropland and grassland. Models were inconsistent among years and species. The potential biotic indicators of landscape and wetland condition examined here would be appropriate for temporal trend analyses, but because of inherent geographic variability would not be appropriate for single-year geographic trend analyses without more extensive evaluations to improve explanatory models.


Oral contraceptive pills, gonadotropin-releasing hormone agonists, or use in combination for treatment of hirsutism: a clinical research center study.
The effectiveness of oral contraceptive pills (OCPs), GnRH agonist (GnRH-a), and a combination of OCPs and GnRH-a in the treatment of hirsute women was compared and the impact of these treatments on hormonal and Ca metabolism was investigated. Thirty-three women were prospectively enrolled and randomized into three treatment groups (11 in each group). The serum levels of LH, estradiol, testosterone, free testosterone, androstenedione, and 17-hydroxyprogesterone declined in all 3 treatment groups, whereas the inclusion of GnRH-a treatment tended to promote a more rapid decrease in these hormone levels. Total cholesterol, low density lipoprotein, and high density lipoprotein levels remained unchanged. The assessment of hirsutism by the Ferriman-Gallwey score revealed a similar 25% reduction in score by all three treatment groups by 6 months. In addition, no difference was detected between groups with respect to hair diameters and the vellus index. Clinical assessment of hirsutism at 3 months by the patients revealed that the GnRH-a and the OCPs-plus-GnRH-a groups had better responses than the group on OCPs alone, but by 6 months all three groups were similar. The symptoms of hot flashes and vaginal dryness were greatest in subjects treated with GnRH-a alone. Serum Ca, phosphorus, alkaline phosphatase, osteocalcin, and 2-h fasting and 24-h urinary Ca excretion levels all increased significantly in subjects treated with the GnRH-a alone, whereas a decrement or no changes occurred for these measurement in the other two groups. The estimated Ca balance was unchanged in the OCPs and the OCPs-plus-GnRH-a groups but declined by 90 mg/day from baseline in the GnRH-a-treated women (p < or = 0.001). Bone density significantly decreased in the lumber spine in women treated with GnRH-a alone, with a less marked decline in the femoral neck. In contrast, women receiving OCPs plus GnRH had increased bone density in the lumbar spine. It is concluded that: 1) clinical measures of hirsutism are not different after 6 months of treatment with OCPs alone, GnRH-a alone, or a combination of the two; 2) the decline in gonadotropins and steroid hormones and improvement in clinical response were more rapid and pronounced when GnRH-a treatment was added to OCP administration; and 3) the negative impact of GnRH-a alone on Ca balance and bone loss limits its usefulness as a single agent for long-term therapy of hirsutism.


Glycosyl-phosphatidylinositol anchor synthesis in paroxysmal nocturnal hemoglobinuria: partial or complete defect in an early step.
The defect in the biosynthesis of the glycosyl-phosphatidyl inositol (GPI) anchor in paroxysmal nocturnal hemoglobinuria (PNH) appears to be in the initial steps. In biosynthetic studies using [3H]mannose, abnormal granulocytes of eight patients, and B lymphocytes transformed by Epstein-Barr virus of six different patients synthesized dolichyl phosphoryl mannose, but little or no later mannosylated intermediates. When fused with murine cell lines known to be deficient at different biosynthetic steps of the GPI anchor, the GPI-anchor-deficient granulocytes of 21/21 patients and lymphocytes from 6/6 patients complemented all murine cell lines except those of class A; cells of this class are not able to add N-acetylglucosamine to phosphatidylinositol. These studies indicate that the defect in GPI-anchor synthesis in PNH is early in the pathway, and is the same as that of class A mutants, but may be partial in some patients, resulting in the production of small amounts of mannosylated intermediates.


On the Rule of Mixtures for Predicting Stress-Softening and Residual Strain Effects in Biological Tissues and Biocompatible Materials.
In this work, we use the rule of mixtures to develop an equivalent material model in which the total strain energy density is split into the isotropic part related to the matrix component and the anisotropic energy contribution related to the fiber effects. For the isotropic energy part, we select the amended non-Gaussian strain energy density model, while the energy fiber effects are added by considering the equivalent anisotropic volumetric fraction contribution, as well as the isotropized representation form of the eight-chain energy model that accounts for the material anisotropic effects. Furthermore, our proposed material model uses a phenomenological non-monotonous softening function that predicts stress softening effects and has an energy term, derived from the pseudo-elasticity theory, that accounts for residual strain deformations. The model's theoretical predictions are compared with experimental data collected from human vaginal tissues, mice skin, poly(glycolide-co-caprolactone) (PGC25 3-0) and polypropylene suture materials and tracheal and brain human tissues. In all cases examined here, our equivalent material model closely follows stress-softening and residual strain effects exhibited by experimental data.


Mucous cell responses in gill and skin of brown trout Salmo trutta fario in acidic, aluminium-containing stream water.
Morphometric examination was carried out on the gills and skin of wild and caged hatchery brown trout Salmo trutta fario in an acidic (pH 4.9 to 5.4; Al 203 to 250 microg l(-1)) and in a non-acidic (pH 6.7 to 7.0; Al 27 to 67 microg l(-1)) stream in the Vosges Mountains (NE France) to assess the sublethal effects of acidic water on the mucous cell response. The caged fish were randomly collected after 2, 4, 7 and 11 d and the wild fish were obtained by electrofishing. After 2 d, a reduction of both mucous cell (MC) number and size was observed in the gills of fish held in the acidic stream, suggesting a massive mucus discharge. Hyperplasia and hypertrophy of cells immediately followed this mucus secretion. In the same fish population, skin examination showed a slight and delayed decrease of MC number but a significant increase of cell size. The number of mucous cells of gills and skin was similar in both wild trout populations, whereas a significant MC hypertrophy was observed in the wild fish of the acidic stream. The present field experiment indicates that caged fish could be useful as early indicators of acidification. In addition, the examination of wild populations suggested the occurrence of adaptive mechanisms, information that might be of importance in the context of river recovery programs.


Ask when--not just whether--it's a risk: How regional context influences local causes of diarrheal disease.
Contemporary epidemiology is enriched when it incorporates ecological concepts about systems and dependencies. With regard to diarrheal disease, the causes of which are many and interacting, the dynamics of within- and between-community disease transmission have distinct components but are also linked in important ways. However, few investigators have studied how regional-scale disease dynamics affect local patterns of diarrheal disease transmission. Characterizing this dependence is important for identifying local- and regional-level transmission pathways. We used data from active surveillance of diarrheal disease prevalence gathered from February 2004 through July 2007 in 21 neighboring Ecuadorian villages to estimate how disease prevalence in spatially and temporally proximate villages modulates the influences of village-level risk and protective factors. We found that the impact of local, village-level interventions such as improved latrines and water treatment can be quite different under conditions of high and low regional disease prevalence. In particular, water treatment was effective only when regional disease prevalence was low, suggesting that person-to-person spread, not waterborne spread, is probably responsible for most between-village transmission in this region. Additional regional-scale data could enhance our understanding of how regional-scale transmission affects local-scale dynamics.


Aggregating sequences that occur in many proteins constitute weak spots of bacterial proteostasis.
Aggregation is a sequence-specific process, nucleated by short aggregation-prone regions (APRs) that can be exploited to induce aggregation of proteins containing the same APR. Here, we find that most APRs are unique within a proteome, but that a small minority of APRs occur in many proteins. When aggregation is nucleated in bacteria by such frequently occurring APRs, it leads to massive and lethal inclusion body formation containing a large number of proteins. Buildup of bacterial resistance against these peptides is slow. In addition, the approach is effective against drug-resistant clinical isolates of Escherichia coli and Acinetobacter baumannii, reducing bacterial load in a murine bladder infection model. Our results indicate that redundant APRs are weak points of bacterial protein homeostasis and that targeting these may be an attractive antibacterial strategy.


Use of Primary Care Services, Care Specialized and Drug Use by Population 65 Years and More in the Community of Madrid, Spain.
Ageing of the Spanish population results in an increase in health services required. Therefore, determine the frequency of the health services utilization in this age group and analyze their determinants has a great interest.The aim was to analyze the utilization of health services among older people living in two urban neighborhoods of northern Madrid. A cross-sectional population-based study. It is studied a cohort of 1327 individuals ≥ 65 years, stratified by age and sex. Nine utilization indicators were defined. For each indicator frequencies and the association of each with the other variables were calculated by multivariate analysis. The distribution of indicators expressed as a percentage of the user population is: GP appoiments/month 64.8% (95%CI 62.3 to 67.4); nursing appoiments/month 44.6% (95% CI 41.2 to 47.2); home medical visits/month 3.1% (95%CI 2.2 to 4.1); home nursing visits/month 3% (95%CI 2.1 to 3.9); hospitalization/year 16.4% (95%CI 14.4 to 18.4); appoiments rheumatologist/orthopedic/year 25.1% (95%CI 22.7 to 27.4); physiotherapist appoiments/year 12.9% (95% CI 11.1 to 14.7); podiatrist appoiments/year 30.6% (95%CI 28.1 to 33.1) and polypharmacy (≥5 drugs) 55.7% (95% CI 53 to 58.4). Comorbidity was the best predictor of health care utilization ranging from OR 4.10 (95%CI: 3.07-5,49) to OR 1.39 (95%CI: 0.97-1.99) in polymedicated and visit the physiotherapist respectively. Cardiovascular disease (OR 1.34; 95%CI 1.03-1,76) and diabetes (OR 1.46; 95%CI: 1.05 -2.02) were independently associated with increased use of family doctor. Dependence was the main determinant for home healthcare (OR 3.38; 95%CI: 1.38-8.28) and nurses (OR 9.71; 95%CI: 4.19-22.48) Mood disorders were associated to polypharmacy (OR 2.06; 95%CI: 1,48-2.86) and to visits to family doctor (OR 1.52; 95%CI: 1,13-2.04). The comorbidity is the strongest predictor of health services utilization. Cardiovascular diseases and diabetes are independently associated to greater use. Dependence is the main determinant of home care. Mood disorders associated with polypharmacy and increased attendances to the General Practitioner.


Prognostic value of tissue protein expression levels of MIB-1 (Ki-67) in Danish ovarian cancer patients. From the 'MALOVA' ovarian cancer study.
The primary objective of this study was to assess the expression of MIB-1 (Ki-67) in tumour tissues from 808 patients with epithelial ovarian tumours. The second was to evaluate, whether MIB-1 (Ki-67) tissue expression levels correlate with clinicopathological parameters and prognosis of the disease. Using tissue arrays (TA), we analysed the MIB-1 (Ki-67) expression levels in tissues from 202 women with borderline ovarian tumours (BOT) (177 stage I, 5 stage II, 19 stage III, 1 stage IV) and 606 ovarian cancer (OC) patients (177 stage I, 64 stage II, 311 stage III, 54 stage IV). Using a 10% cut-off level for MIB-1 (Ki-67) overexpression, 12% of the BOTs and 51% of the OCs were positive for MIB-1 (Ki-67) expression. The frequency of MIB-1 (Ki-67) expression-positive OC increased with increasing FIGO stage (p = 0.003), increasing histological grade (p ≤ 0.0001), and a significantly different distribution of MIB-1 (Ki-67) positive and negative tumours were found in adenocarcinoma NOS, serous adenocarcinomas, mucinous adenocarcinomas, endometrioid adenocarcinomas, non-epithelial and clear-cell carcinomas (p = 0.016). Univariate Kaplan-Meier survival analysis performed on all OC cases showed a significant shorter disease specific survival in patients with positive MIB-1 (Ki-67) expression in the tumour tissue (p ≤ 0.0001). In a Cox survival analysis including 606 FIGO stages I to IV OC cases, FIGO stage (II vs I: HR = 3.00, 95% CI: 1.81-4.99, III-I: HR = 6.41, 95% CI: 3.90-10.50, IV vs I: HR = 12.69, 95% CI: 7.21-22); age at diagnosis pr.10 years (HR = 1.27, 95% CI: 1.15-1.40), residual tumour after surgery (HR = 1.95, 95% CI: 1.40-2.73) and MIB-1 (Ki-67) expression (HR = 1.31, 95% CI: 1.08-1.60) had a significant independent impact on survival. Histological grade (p = 0.14) and histological tumour type (p = 0.35) had no significant independent impact on survival. In conclusion, our results predict that an increased level of MIB-1 (Ki-67) expression in tumour tissue, points to a less favourable outcome for OC patients.


A novel virulence strategy for Pseudomonas aeruginosa mediated by an autotransporter with arginine-specific aminopeptidase activity.
The opportunistic human pathogen, Pseudomonas aeruginosa, is a major cause of infections in chronic wounds, burns and the lungs of cystic fibrosis patients. The P. aeruginosa genome encodes at least three proteins exhibiting the characteristic three domain structure of autotransporters, but much remains to be understood about the functions of these three proteins and their role in pathogenicity. Autotransporters are the largest family of secreted proteins in Gram-negative bacteria, and those characterised are virulence factors. Here, we demonstrate that the PA0328 autotransporter is a cell-surface tethered, arginine-specific aminopeptidase, and have defined its active site by site directed mutagenesis. Hence, we have assigned PA0328 with the name AaaA, for arginine-specific autotransporter of P. aeruginosa. We show that AaaA provides a fitness advantage in environments where the sole source of nitrogen is peptides with an aminoterminal arginine, and that this could be important for establishing an infection, as the lack of AaaA led to attenuation in a mouse chronic wound infection which correlated with lower levels of the cytokines TNFα, IL-1α, KC and COX-2. Consequently AaaA is an important virulence factor playing a significant role in the successful establishment of P. aeruginosa infections.


DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis.
The availability of sequences of entire genomes has dramatically increased the number of protein targets, many of which will need to be overexpressed in cells other than the original source of DNA. Gene synthesis often provides a fast and economically efficient approach. The synthetic gene can be optimized for expression and constructed for easy mutational manipulation without regard to the parent genome. Yet design and construction of synthetic genes, especially those coding for large proteins, can be a slow, difficult and confusing process. We have written a computer program that automates the design of oligonucleotides for gene synthesis. Our program requires simple input information, i.e. amino acid sequence of the target protein and melting temperature (needed for the gene assembly) of synthetic oligonucleotides. The program outputs a series of oligonucleotide sequences with codons optimized for expression in an organism of choice. Those oligonucleotides are characterized by highly homogeneous melting temperatures and a minimized tendency for hairpin formation. With the help of this program and a two-step PCR method, we have successfully constructed numerous synthetic genes, ranging from 139 to 1042 bp. The approach presented here simplifies the production of proteins from a wide variety of organisms for genomics-based studies.


The interaction between acute oligomer Abeta(1-40) and stress severely impaired spatial learning and memory.
In this study, we investigated whether stress can enhance the toxicity of oligomer Abeta(1-40) in the mouse brain. Stress was applied to the animals, consisting of a 2-day inescapable foot shock followed by 3-weekly situation reminders (SRs). We found that stress significantly affected not only the amygdala-dependent (anxiety) but also the hippocampal-dependent (spatial learning and memory) behaviors through the oxidative damage caused in these two regions. However, oligomer Abeta(1-40) treatment alone did not induce behavioral impairment. In addition, combined oligomer Abeta(1-40) and stress treatment increased the glucocorticoid receptor (GR)/mineralocorticoid receptor (MR) ratio and the expression of corticotrophin releasing factor 1 (CRF-1) receptor in the hippocampus. Changes in the components of the hypothalamic-pituitary-adrenal (HPA) axis, such as the GR/MR ratio and CRF-1 level, were observed, accompanied by increasing Abeta accumulation, oxidative stress, nuclear transcription factor (NF-kappaB) hypoactivity, and apoptotic signaling in the hippocampus, and decreasing calbindin D28K and NMDA receptor 2A/2B (NR2A/2B) in the hippocampus, along with alteration of the cholinergic neurons (ChAT) in the medium septum/diagnoid band (MS/DB), noradrenergic neurons (TH) in the locus coeruleus (LC), and serotonergic neurons (5-HT) in the Raphe nucleus. Therefore, apoptosis and synaptic dysfunction in the hippocampus severely induced the impairment of spatial learning and memory. These results suggest that stress may play an important role in the early stages of Alzheimer's disease (AD), and an antioxidant strategy might be a potential therapeutic approach for stress-mediated disorders.


Measurement of GABA in rat brain microdialysates using o-phthaldialdehyde-sulphite derivatization and high-performance liquid chromatography with electrochemical detection.
A number of recent studies have detected gamma-aminobutyric acid (GABA) in brain microdialysates by measuring an electroactive o-phthaldialdehyde (OPA)-alkylthiol derivative using HPLC with electrochemical detection. A particular problem of this approach is the stench of the thiol reagents involved, and the poor stability of the electroactive GABA derivative. Here we report that these practical disadvantages can be overcome by substitution of the OPA-alkylthiol reaction with an OPA-sulphite reaction. This provides a simple, sensitive and reliable means for determination of GABA in microdialysates of rat brain.


Publication Rates in Podium and Poster Presentations at the 2012-2014 American Association of Hip and Knee Surgeons Annual Meetings.
Subspecialty conferences are an important forum for disseminating the latest research relevant to clinical practice. The purpose of this study was to identify publication rates in podium and poster abstracts for the American Association of Hip and Knee Surgeons (AAHKS) Annual Meeting and to identify the most common journals of publication and the reasons for nonpublication. Six hundred ten accepted abstracts (182 podium presentations, 428 posters) from the 2012-2014 AAHKS meetings were searched using Google, Google Scholar, and PubMed. If an abstract could not be found after efforts by multiple searchers, the first author was emailed to determine where the research was published or why it was not published. For articles that were published, the journal, time to publication, and journal impact factor were noted. The overall rate of publication was 71% (436/610). Podium presentations (164/182, 90%) were published at a higher rate than posters (271/428, 63%). The most common journal of publication was the Journal of Arthroplasty (218/436, 50%), followed by Clinical Orthopaedics and Related Research (77/436, 18%) and The Journal of Bone and Joint Surgery (40/436, 9%). Average time to publication was 14.5 months (range, -4 to 44 months) from the date of the conference in which it was presented. Presentations at the AAHKS annual meeting have an impressive rate of publication. The research presented at the meeting is impactful and high quality, warranting consideration for future publication.


Foramina on the zygomatic bone: its clinical significance.
To study the anatomy of zygomatico-orbital, zygomatico-facial and zygomatico temporal foramina in south Indian population. 50 dry adult skulls of south Indian origin were studied. The number of ZO, ZF and ZT were recorded on each side. The distances of these foramina to the nearest palpable bony points, infero-lateral angle of the orbit and infra-orbital foramen were measured using Vernier's calipers. Variable number of ZO and ZF foramina, ranging from 0 to 4 was seen. Single ZO foramen in 56% and 44%, double in 6% and 12%, absent in 38% and 44% on right and left sides were seen. Distance of ZO foramen from inferolateral angle of orbit was 9+2.51 mm and 7.8+1.74 mm on right and left side respectively. The mean distance from infraorbital foramen was 29.93+2.76 mm and 30.19+ 3.52 mm on right and left side respectively. Single ZF foramen in 40% and 42%, double in 10% and 8%, 3 in 4% on right side and 4 in 2% on left sides were seen. In the remaining 46% and 48% of bones, the ZF foramen was absent. Mean distance from inferolateral angle of orbit was 8.05+2.68 and 9.15+2.11 mm respectively on the right and left side. Mean distance from infraorbital foramen was 25.45+4.35 mm and 25.42+4 mm respectively on right and left side. The location of ZO and ZF foramen can be population specific and knowledge of its location is important during stabilization of zygomatic fractures, in endoscopic subperiosteal facelift.


Adding Insult to Injury: Sex, Sexual Orientation, and Juror Decision-Making in a Case of Intimate Partner Violence.
Societal definitions of intimate partner violence (IPV) are highly gendered and heteronormative, resulting in dissonance regarding cases of same-sex IPV. This study explored perceptions of IPV when the context of the case is inconsistent with societal norms regarding sex and sexuality. Mock jurors read a vignette describing a case of alleged IPV in which the sex and sexual orientation of the defendant were manipulated. Participants (N = 415) rendered a verdict and provided ratings of the defendant, victim, and case. Results suggest participants were more confident in a guilty verdict when the defendant was male, compared to female. Further, male defendants were perceived as more morally responsible, but only when the victim was female. Perceptions regarding the crime suggest violence perpetrated by a man against a woman is viewed more adversely than any other condition. Data are discussed in terms of implications for legal decision-makers and public policy.


Randomized controlled study comparing two doses of intravenous granisetron (1 and 3 mg) for acute chemotherapy-induced nausea and vomiting in cancer patients: a non-inferiority trial.
The aim of this study was to assess the non-inferiority of 1 mg to 3 mg granisetron (GRN) injection for the treatment of acute chemotherapy-induced nausea and vomiting (CINV) and to evaluate the tolerability of GRN given at 1 mg in Japanese cancer patients. Patients with cancer receiving highly emetogenic chemotherapy were enrolled in this single-blind randomized controlled study. Patients were randomly assigned to receive GRN at a single dose of 1 or 3 mg. The primary endpoint was the rate of complete protection from emetic events (no vomiting, no retching and no need for rescue medication) during the first 24 h following the initiation of chemotherapy. There were 89 patients in the 1 mg group and 90 patients in the 3 mg group. Complete protection was achieved in 70 patients (78.7%) in the 1 mg group and 73 (81.1%) patients in the 3 mg group. The one-sided test did not reveal non-inferiority of either dose of GRN to the other at a 5% significance level. Our data failed to show the non-inferiority of 1 mg of GRN to 3 mg of GRN administered as a single dose. However, the rate of complete protection from nausea and vomiting was similar in the two groups. Given the recommended dosage in the guidelines and the economic need for reduction of medical care expenses in Japan, prophylactic administration of GRN at 1 mg may be an appropriate, alternative treatment for acute CINV in cancer patients.


Structural convergence between Cryo-EM and NMR reveals intersubunit interactions critical for HIV-1 capsid function.
Mature HIV-1 particles contain conical-shaped capsids that enclose the viral RNA genome and perform essential functions in the virus life cycle. Previous structural analysis of two- and three-dimensional arrays of the capsid protein (CA) hexamer revealed three interfaces. Here, we present a cryoEM study of a tubular assembly of CA and a high-resolution NMR structure of the CA C-terminal domain (CTD) dimer. In the solution dimer structure, the monomers exhibit different relative orientations compared to previous X-ray structures. The solution structure fits well into the EM density map, suggesting that the dimer interface is retained in the assembled CA. We also identified a CTD-CTD interface at the local three-fold axis in the cryoEM map and confirmed its functional importance by mutagenesis. In the tubular assembly, CA intermolecular interfaces vary slightly, accommodating the asymmetry present in tubes. This provides the necessary plasticity to allow for controlled virus capsid dis/assembly.


Export of cellubrevin from the endoplasmic reticulum is controlled by BAP31.
Cellubrevin is a ubiquitously expressed membrane protein that is localized to endosomes throughout the endocytotic pathway and functions in constitutive exocytosis. We report that cellubrevin binds with high specificity to BAP31, a representative of a highly conserved family of integral membrane proteins that has recently been discovered to be binding proteins of membrane immunoglobulins. The interaction between BAP31 and cellubrevin is sensitive to high ionic strength and appears to require the transmembrane regions of both proteins. No other proteins of liver membrane extracts copurified with BAP31 on immobilized recombinant cellubrevin, demonstrating that the interaction is specific. Synaptobrevin I bound to BAP31 with comparable affinity, whereas only weak binding was detectable with synaptobrevin II. Furthermore, a fraction of BAP31 and cellubrevin was complexed when each of them was quantitatively immunoprecipitated from detergent extracts of fibroblasts (BHK 21 cells). During purification of clathrin-coated vesicles or early endosomes, BAP31 did not cofractionate with cellubrevin. Rather, the protein was enriched in ER-containing fractions. When BHK cells were analyzed by immunocytochemistry, BAP31 did not overlap with cellubrevin, but rather colocalized with resident proteins of the ER. In addition, immunoreactive vesicles were clustered in a paranuclear region close to the microtubule organizing center, but different from the Golgi apparatus. When microtubules were depolymerized with nocodazole, this accumulation disappeared and BAP31 was confined to the ER. Truncation of the cytoplasmic tail of BAP31 prevented export of cellubrevin, but not of the transferrin receptor from the ER. We conclude that BAP31 represents a novel class of sorting proteins that controls anterograde transport of certain membrane proteins from the ER to the Golgi complex.


Meniscal suture: a simple method.
A simple technique of meniscal suture is described. It avoids the main problem with most other outside-to-inside suture techniques, which is that knots either have to be left inside the joint or have to be pulled out through the meniscus. These knots often come undone or damage either the joint surface or meniscus. Essentially, a suture is inserted from outside to inside through the meniscus, using a cannulated needle. The end of this suture is then pulled back out through a separate hole in the meniscus, using a suture loop inserted with another cannulated needle. The two ends of the first suture are then tied together.


Nutrition optimization prior to surgery.
Optimization of metabolic state prior to major surgery leads to improved surgical outcomes. Nutrition screening protocols should be implemented in the preoperative evaluation, possibly as part of a bundle. Strategies to minimize hyperglycemia and insulin resistance by aggressive preoperative nutrition and carbohydrate loading may promote maintenance of a perioperative anabolic state, improving healing, reducing complications, and shortening the time to recovery of bowel function and hospital discharge. Short courses of preoperative immune-modulating formulas, using combinations of arginine, ω-3 fatty acids, and other nutrients, have been associated with improved surgical outcomes. These immune-modulating nutrients are key elements of metabolic pathways that promote attenuation of the metabolic response to stress and improve both wound healing and immune function. Patients with severe malnutrition and gastrointestinal dysfunction may benefit from preoperative parenteral nutrition. Continuation of feeding through the intraoperative period for severely stressed hypermetabolic patients undergoing nongastrointestinal surgery is another strategy to optimize metabolic state and reduce prolonged nutrition deficits. In this paper, we review the importance of preoperative nutrition and strategies for effective preoperative nutrition optimization.


Prevalence of Staphylococcus aureus protein A (spa) mutants in the community and hospitals in Oxfordshire.
Staphylococcal protein A (spa) is an important virulence factor which enables Staphylococcus aureus to evade host immune responses. Genotypes known as "spa-types", based on highly variable Xr region sequences of the spa-gene, are frequently used to classify strains. A weakness of current spa-typing primers is that rearrangements in the IgG-binding region of the gene cause 1-2% of strains to be designated as "non-typeable". We developed an improved primer which enabled sequencing of all strains, containing any type of genetic rearrangement, in a large study among community carriers and hospital inpatients in Oxfordshire, UK (6110 isolates). We identified eight novel spa-gene variants, plus one previously described. Three of these rearrangements would be designated "non-typeable" using current spa-typing methods; they occurred in 1.8% (72/3905) asymptomatically carried and 0.6% (14/2205) inpatient S. aureus strains. Some individuals were simultaneously colonized by both formerly non-typeable and typeable strains; previously such patients would have been identified as carrying only currently typeable strains, underestimating mixed carriage prevalence and diversity. Formerly non-typeable strains were found in more spa-types associated with multilocus sequence type ST398 (35%), common among livestock, compared to other groups with any non-typeable strains (1-4%), suggesting particular spa-types may have been under-represented in previous human studies. This improved method allows us to spa-type previously non-typeable strains with rearrangements in the spa-gene and to resolve cases of mixed colonization with deletions in one or more strains, thus accounting for hidden diversity of S. aureus in both community and hospital environments.


Identifying Financially Sustainable Pricing Interventions to Promote Healthier Beverage Purchases in Small Neighborhood Stores.
Residents of low-income communities often purchase sugar-sweetened beverages (SSBs) at small, neighborhood "corner" stores. Lowering water prices and increasing SSB prices are potentially complementary public health strategies to promote more healthful beverage purchasing patterns in these stores. Sustainability, however, depends on financial feasibility. Because in-store pricing experiments are complex and require retailers to take business risks, we used a simulation approach to identify profitable pricing combinations for corner stores. The analytic approach was based on inventory models, which are suitable for modeling business operations. We used discrete-event simulation to build inventory models that use data representing beverage inventory, wholesale costs, changes in retail prices, and consumer demand for 2 corner stores in Baltimore, Maryland. Model outputs yielded ranges for water and SSB prices that increased water demand without loss of profit from combined water and SSB sales. A 20% SSB price increase allowed lowering water prices by up to 20% while maintaining profit and increased water demand by 9% and 14%, for stores selling SSBs in 12-oz cans and 16- to 20-oz bottles, respectively. Without changing water prices, profits could increase by 4% and 6%, respectively. Sensitivity analysis showed that stores with a higher volume of SSB sales could reduce water prices the most without loss of profit. Various combinations of SSB and water prices could encourage water consumption while maintaining or increasing store owners' profits. This model is a first step in designing and implementing profitable pricing strategies in collaboration with store owners.


Application of indirect hemagglutination test and indirect fluorescent antibody test for IgM antibody for diagnosis of melioidosis in Thailand.
In hyperendemic areas such as Thailand, rapid diagnosis of melioidosis depends upon both bacteriological culture and serological methods. However, interpretation of indirect hemagglutination (IHA) for melioidosis which is the only test available, is seriously hampered by increased IHA titers present in one-third to one-half of the population. In order to get the best results from the available tests, IHA and indirect fluorescent antibody for IgM (IFA-IgM) were evaluated in controls and patients in Thailand. IHA titers of greater than or equal to 1:40 were considered remote or recent exposure to P. pseudomallei. IHA titers of this level were found in 47.1% of 227 blood donors and 29.5% of 210 sera submitted for other tests, while IFA-IgM was positive in only one donor who had an IHA titer of 1:1,280. IHA was positive in eight out of nine patients with melioidosis with IHA titers of less than 1:20 to 1:2,560. IFA-IgM was positive in six out of seven melioidosis patients whose sera were available for this test including a serum with IHA titer of less than 1:20. Six patients were predisposed by diabetes mellitus. Among sera serologically tested for melioidosis, 33 had IHA titers of 1:80-1:1,280, 10 of which were positive for IFA-IgM. This study demonstrates high background IHA titers among IHA titers among Thai people which greatly limits its use for serodiagnosis of melioidosis. In sharp contrast, serodiagnosis by IFA-IgM was more successful. Positive IFA-IgM among healthy Thais did exist indicating that serologic tests for melioidosis at best are only supplementary to bacteriological culture and clinical awareness.


Conjugative plasmid from Lactobacillus gasseri LA39 that carries genes for production of and immunity to the circular bacteriocin gassericin A.
Gassericin A is a circular bacteriocin produced by Lactobacillus gasseri strain LA39. We found a 33,333-bp plasmid, designated pLgLA39, in this strain. pLgLA39 contained 44 open reading frames, including seven genes related to gassericin A production/immunity (gaa), as well as genes for replication, plasmid maintenance, and conjugative transfer. pLgLA39 was transferred from LA39 to the type strain of L. gasseri (JCM 1131) by filter mating. The transconjugant exhibited >30-fold-higher more resistance to gassericin A and produced antibacterial activity. Lactobacillus reuteri LA6, the producer of reutericin 6, was proved to harbor a plasmid indistinguishable from pLgLA39 and carrying seven genes 100% identical to gaa. This suggests that pLgLA39 might have been transferred naturally between L. gasseri LA39 and L. reuteri LA6. The seven gaa genes of pLgLA39 were cloned into a plasmid vector to construct pGAA. JCM 1131(T) transformed with pGAA expressed antibacterial activity and resistance to gassericin A. pGAA was segregationally more stable than a pGAA derivative plasmid from which gaaA was deleted and even was more stable than the vector. This suggests the occurrence of postsegregational host killing by the gaa genes. pLgLA39 carried a pemIK homolog, and segregational stabilization of a plasmid by the pLgLA39-type pemIK genes was also confirmed. Thus, pLgLA39 was proved to carry the genes for at least two plasmid maintenance mechanisms, i.e., gaa and pemIK. Plasmids containing a repA gene similar to pLgLA39 repA were distributed in several L. gasseri strains.


Does estrogen adequately protect postmenopausal women against osteoporosis: an iconoclastic perspective.
In sum, the skeletal benefits provided by hormone replacement therapy are important, and estrogen should be justifiably considered one of the fundamentals of menopausal management. However, its efficacy in the prevention of osteoporotic fractures should not be overstated. Low bone mass and fractures remain serious threats in older postmenopausal women, even in the presence of hormone replacement. With the recognition of that reality comes a variety of challenges to clinicians and investigators. Clinicians should use estrogen to its best advantage, though, at the same time, remain vigilant of its limitations. Older postmenopausal women who are, or who have been, taking estrogen should not be a priori considered adequately protected against fracture. The same careful clinical evaluation recommended for the protection of those at increased fracture risk, including bone mass measures, should be available to estrogen-taking women in the later postmenopausal period. Recognizing this need, the National Osteoporosis Foundation has recently recommended bone mineral density testing in older women, regardless of estrogen status, and HCFA reimbursement has become available for this indication. In the presence of low bone density, estrogen-taking women should be afforded appropriate levels of diagnostic and therapeutic attention, with the intent to further reduce fracture risk. Once estrogen therapy has been elected, patients and their clinicians should be aware that bone loss can still be expected in the later postmenopausal years. Periodic reevaluations of bone density and other risks for fracture (e.g. falls) may be appropriate. In the face of continued good health, those reevaluations can be infrequent, but women who have medical conditions associated with adverse skeletal effects should be followed more closely despite their estrogen therapy. If we are to build on the value of estrogen to improve our approach to osteoporosis, additional data are needed. When fracture risk has been considered in complex models that adjust for account other medical and lifestyle variables, a greater protective effect of estrogen has emerged (12), suggesting that there are factors whose actions attenuate those of estrogen. Prominent candidates include genetics, tobacco and alcohol use, medications, body composition, and propensity to fall. There are undoubtedly others yet unidentified. These should be further defined, and the basic mechanisms for interactions between them and estrogen should be examined. It would be clinically advantageous to use these factors to accurately identify not only the women who would benefit from estrogen replacement but also those who would not. Because estrogen has important beneficial effects, an essential research objective should be the development of therapeutic strategies that combine the advantages of estrogen with other modalities (pharmacological or otherwise), to maximally protect the many estrogen-taking women who may be considered at continued risk for fracture. To whit, the initial reports of combination therapy with bisphosphonates are encouraging (13, 14). Other important skeletal agents can be anticipated to interact with estrogen in as-of-yet unforeseen ways. On another front, it can be anticipated that selective estrogen receptor modulators will be subject to estrogen-like limitations when used for osteoporosis prevention/therapy, and their effects will be influenced by a similar, but distinct, array of covariates. Our vision of the role of hormone replacement in the reduction of osteoporotic fracture risk should be considerably sharpened. At one time, estrogen was the lone best hope for the avoidance of fracture, but no longer should we be satisfied with that imperfect solution. The development of much more efficient and effective methods for osteoporosis risk assessment, prevention, and treatment now provides a chance to surpass previous expectations. (ABSTRACT TRUNCATED)


Detection of quantitative trait loci causing abnormal spermatogenesis and reduced testis weight in the small testis (Smt) mutant mouse.
The small testis (Smt) mutant mouse is characterized by a small testis of one third to one half the size of a normal testis, and its spermatogenesis is mostly arrested at early stages of meiosis, although a small number of spermatocytes at the late prophase of meiosis and a few spermatids can sometimes be seen. We performed quantitative trait locus (QTL) analysis of these spermatogenic traits and testis weight using 221 F2 males obtained from a cross between Smt and MOM (Mus musculus molossinus) mice. At the genome-wide 5% level, we detected two QTLs affecting meiosis on chromosomes 4 and 13, and two QTLs for paired testis weight as a percentage of body weight on chromosomes 4 and X. In addition, we found several QTLs for degenerated germ cells and multinuclear giant cells on chromosomes 4, 7 and 13. Interestingly, for cell degeneration, the QTL on chromosome 13 interacted epistatically with the QTL on chromosome 4. These results reveal polygenic participation in the abnormal spermatogenesis and small testis size in the Smt mutant.


A Bayesian Approach for Summarizing and Modeling Time-Series Exposure Data with Left Censoring.
Direct reading instruments are valuable tools for measuring exposure as they provide real-time measurements for rapid decision making. However, their use is limited to general survey applications in part due to issues related to their performance. Moreover, statistical analysis of real-time data is complicated by autocorrelation among successive measurements, non-stationary time series, and the presence of left-censoring due to limit-of-detection (LOD). A Bayesian framework is proposed that accounts for non-stationary autocorrelation and LOD issues in exposure time-series data in order to model workplace factors that affect exposure and estimate summary statistics for tasks or other covariates of interest. A spline-based approach is used to model non-stationary autocorrelation with relatively few assumptions about autocorrelation structure. Left-censoring is addressed by integrating over the left tail of the distribution. The model is fit using Markov-Chain Monte Carlo within a Bayesian paradigm. The method can flexibly account for hierarchical relationships, random effects and fixed effects of covariates. The method is implemented using the rjags package in R, and is illustrated by applying it to real-time exposure data. Estimates for task means and covariates from the Bayesian model are compared to those from conventional frequentist models including linear regression, mixed-effects, and time-series models with different autocorrelation structures. Simulations studies are also conducted to evaluate method performance. Simulation studies with percent of measurements below the LOD ranging from 0 to 50% showed lowest root mean squared errors for task means and the least biased standard deviations from the Bayesian model compared to the frequentist models across all levels of LOD. In the application, task means from the Bayesian model were similar to means from the frequentist models, while the standard deviations were different. Parameter estimates for covariates were significant in some frequentist models, but in the Bayesian model their credible intervals contained zero; such discrepancies were observed in multiple datasets. Variance components from the Bayesian model reflected substantial autocorrelation, consistent with the frequentist models, except for the auto-regressive moving average model. Plots of means from the Bayesian model showed good fit to the observed data. The proposed Bayesian model provides an approach for modeling non-stationary autocorrelation in a hierarchical modeling framework to estimate task means, standard deviations, quantiles, and parameter estimates for covariates that are less biased and have better performance characteristics than some of the contemporary methods.


Harnessing the evolvability of tricyclic microviridins to dissect protease-inhibitor interactions.
Understanding and controlling proteolysis is an important goal in therapeutic chemistry. Among the natural products specifically inhibiting proteases microviridins are particularly noteworthy. Microviridins are ribosomally produced and posttranslationally modified peptides that are processed into a unique, cagelike architecture. Here, we report a combined rational and random mutagenesis approach that provides fundamental insights into selectivity-conferring moieties of microviridins. The potent variant microviridin J was co-crystallized with trypsin, and for the first time the three-dimensional structure of microviridins was determined and the mode of inhibition revealed.


Cytokine-induced differentiation of multipotent adult progenitor cells into functional smooth muscle cells.
Smooth muscle formation and function are critical in development and postnatal life. Hence, studies aimed at better understanding SMC differentiation are of great importance. Here, we report that multipotent adult progenitor cells (MAPCs) isolated from rat, murine, porcine, and human bone marrow demonstrate the potential to differentiate into cells with an SMC-like phenotype and function. TGF-beta1 alone or combined with PDGF-BB in serum-free medium induces a temporally correct expression of transcripts and proteins consistent with smooth muscle development. Furthermore, SMCs derived from MAPCs (MAPC-SMCs) demonstrated functional L-type calcium channels. MAPC-SMCs entrapped in fibrin vascular molds became circumferentially aligned and generated force in response to KCl, the L-type channel opener FPL64176, or the SMC agonists 5-HT and ET-1, and exhibited complete relaxation in response to the Rho-kinase inhibitor Y-27632. Cyclic distention (5% circumferential strain) for 3 weeks increased responses by 2- to 3-fold, consistent with what occurred in neonatal SMCs. These results provide evidence that MAPC-SMCs are phenotypically and functionally similar to neonatal SMCs and that the in vitro MAPC-SMC differentiation system may be an ideal model for the study of SMC development. Moreover, MAPC-SMCs may lend themselves to tissue engineering applications.