Standard regression-based methods for measuring recovery after sport-related concussion.
Clinical decision making about an athlete's return to competition after concussion is hampered by a lack of systematic methods to measure recovery. We applied standard regression-based methods to statistically measure individual rates of impairment at several time points after concussion in college football players. Postconcussive symptoms, cognitive functioning, and balance were assessed in 94 players with concussion (based on American Academy of Neurology Criteria) and 56 noninjured controls during preseason baseline testing, and immediately, 3 hr, and 1, 2, 3, 5, and 7 days postinjury. Ninety-five percent of injured players exhibited acute concussion symptoms and impairment on cognitive or balance testing immediately after injury, which diminished to 4% who reported elevated symptoms on postinjury day 7. In addition, a small but clinically significant percentage of players who reported being symptom free by day 2 continued to be classified as impaired on the basis of objective balance and cognitive testing. These data suggest that neuropsychological testing may be of incremental utility to subjective symptom checklists in identifying the residual effects of sport-related concussion. The implementation of neuropsychological testing to detect subtle cognitive impairment is most useful once postconcussive symptoms have resolved. This management model is also supported by practical and other methodological considerations.


Detection and Classification of Sleep Apnea and Hypopnea Using PPG and SpO 2 Signals.
In this work, a detection and classification method for sleep apnea and hypopnea, using photopletysmography (PPG) and peripheral oxygen saturation (SpO 2) signals, is proposed. The detector consists of two parts: one that detects reductions in amplitude fluctuation of PPG (DAP)and one that detects oxygen desaturations. To further differentiate among sleep disordered breathing events (SDBE), the pulse rate variability (PRV) was extracted from the PPG signal, and then used to extract features that enhance the sympatho-vagal arousals during apneas and hypopneas. A classification was performed to discriminate between central and obstructive events, apneas and hypopneas. The algorithms were tested on 96 overnight signals recorded at the UZ Leuven hospital, annotated by clinical experts, and from patients without any kind of co-morbidity. An accuracy of 75.1% for the detection of apneas and hypopneas, in one-minute segments,was reached. The classification of the detected events showed 92.6% accuracy in separating central from obstructive apnea, 83.7% for central apnea and central hypopnea and 82.7% for obstructive apnea and obstructive hypopnea. The low implementation cost showed a potential for the proposed method of being used as screening device, in ambulatory scenarios.


Initial genomic scan of the NIMH genetics initiative bipolar pedigrees: chromosomes 3, 5, 15, 16, 17, and 22.
As part of the four-center NIMH Genetics Initiative on Bipolar Disorder we carried out a genomic scan of chromosomes 3, 5, 15, 16,17, and 22. Genotyping was performed on a set of 540 DNAs from 97 families, enriched for affected relative pairs and parents where available. We report here the results of the initial 74 markers that have been typed on this set of DNAs. The average distance between markers (theta) was 12.3 cM. Nonparametric analysis of excess allele sharing among affected sibling pairs used the SIBPAL program of the S.A.G.E. package to test three hierarchical models of affected status. D16S2619 gave some evidence of linkage to bipolar disorder, with P = 0.006 for Model II (in which bipolar 1, bipolar 2 and schizoaffective-bipolar type individuals are considered affected). Nearby markers also showed increased allele sharing. A second interesting region was toward the telomere of chromosome 5q, where D5S1456 and nearby markers showed increased allele sharing; for D5S1456, P = 0.05, 0.015 and 0.008 as the models of affected status become more broad. MOD score analysis also supported the possible presence of a susceptibility locus in this region of chromosome 5. A pair of adjacent markers on chromosome 3, D3S2405 and D3S3038, showed a modest increased allele sharing in the broad model. Several isolated markers had excess allele sharing at the P < 0.05 level under a single model. D15S217 showed a MOD score of 2.37 (P < 0.025). Multipoint analysis flagged the region of chromosome 22 around D22S533 as the most interesting. Thus, several regions showed modest evidence for linkage to bipolar disorder in this initial genomic scan of these chromosomes, including broad regions near previous reports of possible linkage.


Sclerosing cholangitis.
Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by strictures of the biliary tree. It is closely associated with inflammatory bowel disease, particularly ulcerative colitis, although the precise etiology remains unknown. The prevalence of PSC, with its attendant medical burdens, appears to be higher than previously estimated; consequently, increasing numbers of patients are being diagnosed and treated in secondary care. Research into etiopathogenesis, epidemiology, small-duct PSC, and associations with cholangiocarcinoma and colorectal carcinoma are discussed, along with medical therapy and transplantation. Small-duct PSC is a distinct clinical entity associated with a benign course and minimal risk of cholangiocarcinoma compared with large-duct PSC. The incidence of gall bladder cancer is increased in patients with PSC, and yearly ultrasonographic screening, with cholecystectomy for gall bladder polyps, is recommended. Recent pilot studies have suggested that high-dose ursodeoxycholic acid (20 mg/kg body weight) may slow liver fibrosis and cholangiographic appearances. Importantly, ursodeoxycholic acid has also been shown to reduce the incidence of colonic dysplasia and colorectal cancer in patients with PSC and ulcerative colitis. Endoscopic therapy, usually balloon dilatation, is effective for benign dominant strictures that are causing jaundice. Liver transplant remains the only treatment in end-stage disease; however, posttransplant recurrence of PSC occurs in as many as one third of patients. Colorectal cancer develops posttransplant in 5 to 10% of those with inflammatory bowel disease. Annual screening colonoscopy is recommended for all patients with PSC with coexisting inflammatory bowel disease.


The relations between prepotential, resting potential, and latent period in frog muscle fibers.
1. Prepotentials and action potentials were recorded from amphibian striated muscle fibers. Intracellular electrodes were used for stimulating and recording. The resting potential was varied from 55 to 120 mv. by alterations of the KCl concentration of the Ringer's fluid. The magnitude of the prepotential at the initiation of the spike potential was measured and compared to the resting potential and the latent period (time between stimulus "make" and excitation). The magnitude of this prepotential varied with the resting potential. 2. A large prepotential or cathodal depolarization was required to excite a fiber with a high resting potential. If a fiber with a high resting potential fired late (long latency), the adequate prepotential was larger than if the fiber fired early. Fibers with low resting potentials had smaller adequate prepotentials. Also, the adequate prepotential was independent of the latent period, in these depolarized fibers. 3. If the concentration of Ca(++) was increased tenfold, the adequate prepotential of depolarized fibers became strongly dependent upon the latency. 4. Fibers with large or normal resting potentials were prone to respond repetitively during the passage of long duration shock, whereas depolarized and Ca(++)-treated fibers were not. 5. The so-called critical membrane potential (which is defined as the transmembrane potential at the point of excitation) was not independent of the resting potential.


Does partial replacement of corn with glycerin in beef cattle diets affect in vitro ruminal fermentation, gas production kinetic, and enteric greenhouse gas emissions?
Five in vitro experiments were conducted with the following objectives: 1) To evaluate the ruminal fermentation of three different single ingredients: corn, glycerin, and starch (Exp. 1 and 2); 2) To assess effects of partially replacing corn with glycerin in beef cattle diets on ruminal fermentation pattern (Exp. 3 and 4); and 3) To evaluate the effects of glycerin inclusion on the extension of ruminal DM digestibility of feeds with high (orchard hay) and low (corn) fiber content (Exp. 5). For Exp. 1 and 2, two in vitro systems (24-bottle AnkomRF and 20-serum bottles) were used in four consecutive fermentation batches to evaluate gas production (GP), fermentation profiles, enteric methane (CH4), and carbon dioxide (CO2) of corn, glycerin, and starch. The 24 h total GP, acetate concentration, and acetate: propionate ratio decreased only when glycerin was added to the diet (P < 0.01). The 48-h total GP and metabolizable energy were greatest for corn (P < 0.01), and similar between glycerin and starch. The starch treatment had the lowest total volatile fatty acids concentration (P = 0.01). Glycerin had greatest CH4 production, lag time, and maximum gas volume of the first pool (P < 0.05). However, the maximum gas volume of the second pool was greatest for corn (P < 0.05), and similar between glycerin and starch. The starch treatment had the greatest specific rates of digestion for first and second pools (P < 0.05). Production of CO2 (mL/g) was greater for corn (P < 0.01), but similar for glycerin and starch. For Exp. 3 and 4, the same systems were used to evaluate four different levels of glycerin [0, 100, 200, and 300 g/kg of dry matter (DM)] replacing corn in beef cattle finishing diets. Glycerin levels did not affect 24 and 48 h total GP, CH4, and CO2 (P > 0.05). The inclusion of glycerin linearly decreased acetate concentration (P = 0.03) and acetate: propionate ratio (P = 0.04). For Exp. 5, two DaisyII incubators were used to evaluate the in vitro dry matter digestibility (IVDMD) of the following treatments: orchard hay; corn; orchard hay + glycerin; and corn + glycerin. Glycerin inclusion decreased orchard hay IVDMD (P < 0.01) but did not affect corn IVDMD (P > 0.05). We concluded that, under these experimental conditions, glycerin has similar energy efficiency when used in replacement of corn and included at up to 300 g/kg in beef cattle diets.


Effects of Metal Ions on Conductivity and Structure of Single DNA Molecule in Different Environmental Conditions.
We design a novel nano-gap electrode to measure the current of DNA molecule, by which the current-voltage characteristics of individual native DNA, Ag-DNA and Ni-DNA molecules are obtained, respectively. The results show that the voltage gap of Ag- and Ni-DNA is higher than that of native DNA, and the conductance is lower than native DNA in neutral environment. The structure transition from B- to Z-DNA is observed in the presence of high concentrations of nickel ions and Ag-DNA appears chaos state by STM image and U-V spectra characterization. But in alkaline environment, the conductance of Ni-DNA rises and the voltage gap decreases with the increasing of nickel ion concentration denotes that the conductive ability of Ni-DNA is higher than that of native DNA.


Comparison of a short irradiation (50 sec) by different wavelengths on audiogenic seizures in magnesium-deficient mice: evidence for a preventive neuroprotective effect of yellow.
Audiogenic seizures triggered by an acoustic stimulus of determined frequency and amplitude have been described in many laboratory animals in many circumstances including magnesium deficiency. This model, recently validated, was used, in DBA/2 mice, to study the preventive neuroprotective effect of 6 wavelengths of the visible spectrum used in Chromatotherapia* (lambda(max) 440, 484, 528, 572, 616 and 660 nm) at low irradiance. Each short illumination lasted 50 seconds and was followed by 20 minutes of darkness. It appeared that yellow fully protected 16 out of 17 mice from seizure occurrence. Green allowed the survival of 69% of mice but did not protect them from seizure occurrence. On the contrary, the other four colors (orange, red, purple and blue) failed to protect the mice and showed a tendency to accelerate their death. White color was not protective but allowed the difficult survival of 30% of mice. Darkness had no protective effect. These results even though surprising open a great field of investigation.


Antibiotic resistance in Campylobacter: emergence, transmission and persistence.
Campylobacter is a leading foodborne bacterial pathogen, which causes gastroenteritis in humans. This pathogenic organism is increasingly resistant to antibiotics, especially fluoroquinolones and macrolides, which are the most frequently used antimicrobials for the treatment of campylobacteriosis when clinical therapy is warranted. As a zoonotic pathogen, Campylobacter has a broad animal reservoir and infects humans via contaminated food, water or milk. Antibiotic usage in both animal agriculture and human medicine, can influence the development of antibiotic-resistant Campylobacter. This review will describe the trend in fluoroquinolone and macrolide resistance in Campylobacter, summarize the mechanisms underlying the resistance to various antibiotics and discuss the unique features associated with the emergence, transmission and persistence of antibiotic-resistant Campylobacter. Special attention will be given to recent findings and emphasis will be placed on Campylobacter resistance to fluoroquinolones and macrolides. A future perspective on antibiotic resistance and potential approaches for the control of antibiotic-resistant Campylobacter, will also be discussed.


Promoting success or preventing failure: cultural differences in motivation by positive and negative role models.
In two studies, cross-cultural differences in reactions to positive and negative role models were examined. The authors predicted that individuals from collectivistic cultures, who have a stronger prevention orientation, would be most motivated by negative role models, who highlight a strategy of avoiding failure; individuals from individualistic cultures, who have a stronger promotion focus, would be most motivated by positive role models, who highlight a strategy of pursuing success. In Study 1, the authors examined participants' reported preferences for positive and negative role models. Asian Canadian participants reported finding negative models more motivating than did European Canadians; self-construals and regulatory focus mediated cultural differences in reactions to role models. In Study 2, the authors examined the impact of role models on the academic motivation of Asian Canadian and European Canadian participants. Asian Canadians were motivated only by a negative model, and European Canadians were motivated only by a positive model.


From theory to practice: improving the impact of health services research.
While significant strides have been made in health research, the incorporation of research evidence into healthcare decision-making has been marginal. The purpose of this paper is to provide an overview of how the utility of health services research can be improved through the use of theory. Integrating theory into health services research can improve research methodology and encourage stronger collaboration with decision-makers. Recognizing the importance of theory calls for new expectations in the practice of health services research. These include: the formation of interdisciplinary research teams; broadening the training for those who will practice health services research; and supportive organizational conditions that promote collaboration between researchers and decision makers. Further, funding bodies can provide a significant role in guiding and supporting the use of theory in the practice of health services research. Institutions and researchers should incorporate the use of theory if health services research is to fulfill its potential for improving the delivery of health care.


Effect of oral intake of choline-stabilized orthosilicic acid on hair tensile strength and morphology in women with fine hair.
The appearance of hair plays an important role in people's overall physical appearance and self-perception. Silicon (Si) has been suggested to have a role in the formation of connective tissue and is present at 1-10 ppm in hair. Choline-stabilized orthosilicic acid ("ch-OSA") is a bioavailable form of silicon which was found to improve skin microrelief and skin mechanical properties in women with photoaged skin. The effect of ch-OSA on hair was investigated in a randomized, double blind, placebo-controlled study. Forty-eight women with fine hair were given 10 mg Si/day in the form of ch-OSA beadlets (n = 24) or a placebo (n = 24), orally for 9 months. Hair morphology and tensile properties were evaluated before and after treatment. Urinary silicon concentration increased significantly in the ch-OSA supplemented group but not in the placebo group. The elastic gradient decreased in both groups but the change was significantly smaller in the ch-OSA group (-4.52%) compared to placebo group (-11.9%). Break load changed significantly in the placebo group (-10.8%) but not in the ch-OSA supplemented group (-2.20%). Break stress and elastic modulus decreased in both groups but the change was smaller in the ch-OSA group. The cross sectional area increased significantly after 9 months compared to baseline in ch-OSA supplemented subjects but not in the placebo group. The change in urinary silicon excretion was significantly correlated with the change in cross sectional area. Oral intake of ch-OSA had a positive effect on tensile strength including elasticity and break load and resulted in thicker hair.


Myasthenia Gravis Attack after Oral Risperidone Treatment: A Case Report.
Myasthenia gravis (MG) is an autoimmune disease, which can be triggered by anticholinergic agents. The 6-year-old female patient was admitted to the outpatient clinic. She was had been previously diagnosed with comorbid attention deficit hyperactivity disorder and conduct disorder and was receiving short-acting methylphenidate and risperidone, as recommended by a child psychiatrist. However, after using the drugs, she stated that she was overly tired during the day and that her eyelids drooped. Hence, the current treatment was stopped. She was hospitalized with a prediagnosis of ocular type MG and pyridostigmine (90 mg/day) treatment was started. The patient recovered and subsequently the treatment was stopped. Since psychiatric symptoms of the patient resurfaced, long-acting methylphenidate treatment was initiated. During this treatment, the symptoms of MG did not return. The Naranjo's scale of adverse drug reaction probability was completed. Consequently, there may be an association between risperidone and MG.


An improved pairwise decomposable finite-difference Poisson-Boltzmann method for computational protein design.
Our goal is to develop accurate electrostatic models that can be implemented in current computational protein design protocols. To this end, we improve upon a previously reported pairwise decomposable, finite difference Poisson-Boltzmann (FDPB) model for protein design (Marshall et al., Protein Sci 2005, 14, 1293). The improvement involves placing generic sidechains at positions with unknown amino acid identity and explicitly capturing two-body perturbations to the dielectric environment. We compare the original and improved FDPB methods to standard FDPB calculations in which the dielectric environment is completely determined by protein atoms. The generic sidechain approach yields a two to threefold increase in accuracy per residue or residue pair over the original pairwise FDPB implementation, with no additional computational cost. Distance dependent dielectric and solvent-exclusion models were also compared with standard FDPB energies. The accuracy of the new pairwise FDPB method is shown to be superior to these models, even after reparameterization of the solvent-exclusion model.


Networking Hospital ePrescribing: A Systemic View of Digitalization of Medicines' Use in England.
Medicine management is at the core of hospital care and digitalization of prescribing and administration of medicines is often the focus of attention of health IT programs. This may be conveyed to the public in terms of the elimination of paper-based drug charts and increased readability of doctors' prescriptions. Based on analysis of documents about hospital medicines supply and use (including systems' implementation) in the UK, in this conceptual paper electronic prescribing and administration are repositioned as only one aspect of an important wider transformation in medicine management in hospital settings, involving, for example, procurement, dispensing, auditing, waste management, research and safety vigilance. Approaching digitalization from a systemic perspective has the potential to uncover the wider implications of this transformation for patients, the organization and the wider health care system.


Complete Genome Sequence of Enterococcus hirae R17, a Daptomycin-Resistant Bacterium Isolated from Retail Pork in China.
Daptomycin-resistant Enterococcus hirae R17 was isolated from retail pork sold at a free-trade market in Beijing, China. The complete genome sequence of R17 contains a circular 2,886,481-bp chromosome and a circular 73,574-bp plasmid. Genes involved in cell envelope homeostasis of this bacterium were identified by whole-genome analysis.


Occupational risk factors for female breast cancer: a review.
Although progress has been made in identifying personal risk factors and in improving treatment for female breast cancer, incidence rates continue to increase. With women now occupying a sizable fraction of the workforce, it is worth inquiring whether there are occupational risk factors for breast cancer. This is a review of occupational studies on female breast cancer. Suitable reports and published articles with associations of female breast cancer and occupation were identified from technical reports, by searching the MEDLINE bibliographic data base, and by reviewing each paper on cancer that was published in 20 major journals during the period from about 1971-94. A total of 115 studies were identified; 19 studies relied exclusively on data collected for administrative purposes, and there were four incident case-control studies and 92 cohort studies. Although data for individual industries, occupations, and exposures were sparse, there was limited evidence of an association with employment in the pharmaceutical industry and among cosmetologists and beauticians. Associations were also found for chemists and occupations with possible exposure to extremely low frequency electromagnetic fields, but potential methodological weaknesses preclude drawing any definite conclusions. There was little support for increased risks among textiles workers, dry cleaning workers, and nuclear industry workers. Few high quality occupational studies directed specifically toward women have been carried out to allow the unambiguous identification of occupational risk factors for breast cancer. It is suggested that investigations that account for non-occupational risk factors and that assess exposure in a more detailed way be carried out. One strategy already suggested is to conduct population based, case-control studies in which subjects are interviewed about their occupational histories and exposure to chemical and physical agents which are then attributed from the job descriptions by a team of experts. These studies can then be supplemented when necessary with cohort studies of specific populations.


Bomb blast injuries: Tertiary care hospital in-patient experience over the last 20 years.
To examine the distribution and nature of limb injuries in young bomb blast victimsand their management in a tertiary healthcare setting. The retrospective study was conducted at the Aga Khan University Hospital, Karachi and data was searched using the University Medical Record Database with International Classification of Diseases-Ninth Revision-Clinical Modification codes for injuries and terrorism between 1990 and 2012. Data regarding injuries, including admission time, hospital stay duration, interventions and mortality, was collected and classified as per the New Injury Severity Score. There were 22 patients in the study; 19(86.3%) males and 3(13.7%) females. The mean age of the sample was 13.1±4.1 years. Median length of hospital stay was 9 days (range: 2-42 days). Sixteen (72.7%) patients required operative intervention. Patients rarely had accompanying injuries with limb injuries. Four (18%) patients needed open reduction and internal fixation. Two (9%) patients needed open reduction and external fixation. One (4.5%) patient required a limb amputation. One (4.5%) patient required ileal resection with ileoileal anastomosis due to shrapnel perforations. There was no mortality. The young age group was not severely affected by limb trauma. Injuries sustained were infrequently accompanied by severe/critical injury severity scores.


The Mouse Tumor Biology Database (MTB): a central electronic resource for locating and integrating mouse tumor pathology data.
The Mouse Tumor Biology Database (MTB) is designed to provide an electronic data storage, search, and analysis system for information on mouse models of human cancer. The MTB includes data on tumor frequency and latency, strain, germ line, and somatic genetics, pathologic notations, and photomicrographs. The MTB collects data from the primary literature, other public databases, and direct submissions from the scientific community. The MTB is a community resource that provides integrated access to mouse tumor data from different scientific research areas and facilitates integration of molecular, genetic, and pathologic data. Current status of MTB, search capabilities, data types, and future enhancements are described in this article.


Prevalence of malnutrition and its correlates in older adults living in long stay institutions situated in Beirut, Lebanon.
Malnutrition represents an important issue in older adults; unfortunately, there is lack of data concerning this topic in Lebanon. This paper aims to provide a description of nutritional status and its correlates in older adults living in long stay institutions situated in Beirut. This cross-sectional study was conducted in three long stay institutions in Beirut in 2012. The study population was composed of people aged 65 years and above, having a score of Folstein Mini Mental State Examination (MMSE) greater than 14 and without renal failure requiring dialysis. Subjects meeting inclusion criteria filled out a questionnaire consisting of nutritional status scale (Mini Nutritional Assessment: MNA) and several other parts (demographic, self-assessment of the state health, smoking and alcohol, physical dependence, quality of life, frailty, depression, social isolation and loneliness). Data were entered and analyzed using the statistical software SPSS (Statistical Package for Social Sciences), version 17.0 (Chicago, IL, USA). Among 111 older adults (55 men and 56 women), 14 (12.6%) were malnourished, 54 (48.7%) were at risk of malnutrition and 43 (38.7%) had an adequate nutritional status. Multivariate analysis showed that physical exercise, depression, frailty and cognitive function were independent correlates of nutritional status of older adults. This model explained 42.2% (adjusted R2 = 0.422) of the older adults nutritional status variability. We found a moderate percentage of malnutrition in older adults living in long stay institutions situated in Beirut, and the correlates of malnutrition in older adults were low physical exercise, depression, frailty and low cognitive function.


Array CGH identifies distinct DNA copy number profiles of oncogenes and tumor suppressor genes in chromosomal- and microsatellite-unstable sporadic colorectal carcinomas.
DNA copy number changes represent molecular fingerprints of solid tumors and are as such relevant for better understanding of tumor development and progression. In this study, we applied genome-wide array comparative genomic hybridization (aCGH) to identify gene-specific DNA copy number changes in chromosomal (CIN)- and microsatellite (MIN)-unstable sporadic colorectal cancers (sCRC). Genomic DNA was extracted from microdissected, matching normal colorectal epithelium and invasive tumor cells of formalin-fixed and paraffin-embedded tissues of 22 cases with colorectal cancer (CIN = 11, MIN = 11). DNA copy number changes were determined by aCGH for 287 target sequences in tumor cell DNAs, using pooled normal DNAs as reference. aCGH data of tumor cell DNAs was confirmed by fluorescence in situ hybridization (FISH) for three genes on serial tissues as those used for aCGH. aCGH revealed DNA copy number changes previously described by metaphase CGH (gains 7, 8q, 13q, and 20q; losses 8p, 15q, 18q, and 17p). However, chromosomal regions 20q, 13q, 7, and 17p were preferentially altered in CIN-type tumors and included DNA amplifications of eight genes on chromosome 20q (TOP1, AIB1, MYBL2, CAS, PTPN1, STK15, ZNF217, and CYP24), two genes on chromosome 13q (BRCA2 and D13S25), and three genes on chromosome 7 (IL6, CYLN2, and MET) as well as DNA deletions of two genes on chromosome 17p (HIC1 and LLGL1). Finally, additional CIN-tumor-associated DNA amplifications were identified for EXT1 (8q24.11) and MYC (8q24.12) as well as DNA deletions for MAP2K5 (15q23) and LAMA3 (18q11.2). In contrast, distinct MIN-tumor-associated DNA amplifications were detected for E2F5 (8p22-q21.3), GARP (11q13.5-q14), ATM (11q22.3), KAL (Xp22.3), and XIST (Xq13.2) as well as DNA deletions for RAF1 (3p25), DCC (18q21.3), and KEN (21q tel). aCGH revealed distinct DNA copy number changes of oncogenes and tumor suppressor genes in CIN- and MIN-type sporadic colorectal carcinomas. The identified candidate genes are likely to have distinct functional roles in the carcinogenesis and progression of CIN- and MIN-type sporadic CRCs and may be involved in the differential response of CIN- and MIN-type tumor cells to (adjuvant) therapy, such as 5-fluorouracil.


Plasma Vitamin D (25-Hydroxyvitamin D) Levels in Hispaniolan Amazon Parrots (Amazona ventralis) Housed Indoors Over Time.
Vitamin D is a hormone that can be ingested or synthesized in the body when skin is exposed to ultraviolet radiation (UV), typically from sunlight. In captivity, birds with no sunlight exposure may develop vitamin D deficiencies that may contribute to hypocalcemic conditions, even when fed a diet supplemented with vitamin D. An initial pilot study with Hispaniolan Amazon parrots (Amazona ventralis) conducted approximately 18 mo prior indicated there were significant differences in the 25-hydroxyvitamin D (vit D) plasma levels between the resident parrots (indoor-only housing) and a new group of historically outdoor-housed parrots (new parrots) 5 days after the arrival of the latter at our institution. The goals of this study were to determine if vit D, ionized calcium (Ca2+), and ionized magnesium (Mg2+) levels changed from baseline values (taken 18 mo prior) in the new birds as well as to compare those values to those of the resident birds over time. The treatment was a change in husbandry for the new parrots (no UV exposure and diet as provided for the resident parrots). To accomplish this, the authors compared vit D levels in the same two groups of birds that were fed the same vit D-fortified diet and given no access to natural or artificial UV light exposure for 18 mo. The resident parrots (N = 9) had been housed indoors for approximately 20 yr with no exposure to natural or supplemented UV light. The second group of birds (new parrots; N = 8) had been housed outdoors prior to the initiation of the pilot study in 2016 and were fed a similar-but not identical-diet prior to their arrival. Plasma samples were sent to the Michigan State University Diagnostic Center for Population and Animal Health for analysis. Test results demonstrated differences between the two groups of parrots, largely attributed to the decrease in vit D plasma levels in the new parrots over time to values equivalent to those measured in the resident birds. Differences were seen in plasma Ca2+, while no differences were demonstrated relative to Mg2+. We discuss these findings and suggest that plasma vit D levels decrease in the absence of UV light, even when animals are maintained on a vit D-fortified and balanced diet.


Foodinformatics: Quantitative Structure-Property Relationship Modeling of Volatile Organic Compounds in Peppers.
The aim of this work was the foodinformatic (chemoinformatic) modeling of volatile organic compounds (VOCs) of different samples of peppers based on a quantitative structure-property relationship (QSPR) for the retention indices of 273 identified compounds. The experimental retention indices were measured by means of comprehensive two-dimensional gas chromatography combined with quadrupole-mass spectrometry (GC × GC/qMS) using the BPX5 and BP20 column coupled system. All the VOCs were represented by means of both conformation-independent molecular descriptors and molecular fingerprints calculated in the Dragon and PaDEL-Descriptor software. The dataset was divided into training, validation and test sets of molecules according to the Balanced Subsets Method (BSM). Subsequently, the V-WSP unsupervised variable reduction method was used to reduce the presence of multicollinearity, redundancy, and noise in the initial pool of 4,336 molecular descriptors and fingerprints. Using this method, a reduced pool of 1,664 was submitted to the supervised selection by means of the replacement method (RM) variable subset selection in order to define a four-descriptor model. The quality of the model was measured by means of the coefficient of determination and the root-mean-square deviation in fitting ( R train 2 = 0 . 879 and RMSD train = 72.1 ), validation ( R val 2 = 0 . 832 and RMSD val = 91.7 ), and prediction ( R test 2 = 0 . 915 and RMSD test = 55.4 ). The negligible differences among the parameters in the three sets indicate a stable and predictive QSPR model. This quantitative structure-activity relationship was developed keeping in mind the five principles defined by the Organization for Economic Co-operation and Development (OECD) to make it applicable. PRACTICAL APPLICATION: This predictive mathematical model developed from the retention indices of 273 volatile organic compounds (VOCs) detected in pepper samples could be useful for chromatographers working on the identification of other common VOCs in peppers or other foods by means of comprehensive two-dimensional gas chromatography combined with quadrupole-mass spectrometry (GC × GC/qMS) using a bi-dimensional stationary phase coupled system (BPX5 and BP20).


Rehabilitation after traumatic brain injury.
Traumatic brain injury (TBI) is a growing problem in the US, with significant morbidity and economic implications. This diagnosis spans a wide breath of injuries from concussion to severe TBI. Thus, rehabilitation is equally diverse in its treatment strategies targeting those symptoms that are functionally limiting with the ultimate goal of independence and community reintegration. In severe TBI, rehabilitation can be lifelong. Acute care rehabilitation focuses on emergence from coma and prognostication of recovery. Therapeutic modalities and exercise, along with pharmacologic intervention, can target long-term motor and cognitive sequelae. Complications of severe TBI that are functionally limiting and impede therapy include heterotopic ossification, agitation, dysautonomia, and spasticity. In mild TBI, most patients recover quickly but education on repeat exposure is imperative, with the implications of consecutive injuries being potentially devastating. Furthermore, rehabilitation targets lingering symptoms including sleep disturbance, visuospatial deficits, headaches, and cognitive dysfunction. As research on the entire TBI population improves, commonalities in the disease process may emerge, helping rationalize therapeutic interventions and providing more robust targets for treatment.


The stress of stigma: exploring the effect of weight stigma on cortisol reactivity.
To determine the physiological impact of exposure to weight stigma by examining alterations in salivary cortisol among lean and overweight women. Participants were 123 lean and overweight adult women (mean body mass index = 26.99 [7.91] kg/m(2)). Participants' salivary cortisol was assessed both before and after either a weight stigmatizing or a neutral video. Participants completed self-report measures of mood and reactions to the video. Height and weight were obtained at the conclusion of the study. Participants in the stigmatizing condition exhibited significantly greater cortisol reactivity when compared with those in the neutral condition, irrespective of weight status (Pillai trace = 0.077; F(1,85) = 7.22, p = .009). Lean and overweight women in the stigmatizing condition were equally likely to find the video upsetting and were equally likely to report that they would rather not see obese individuals depicted in a stigmatizing manner in the media. Exposure to weight-stigmatizing stimuli was associated with greater cortisol reactivity among lean and overweight women. These findings highlight the potentially harmful physiological consequences of exposure to weight stigma.


Single-cell sequencing in hematology.
In this review, we highlight key recent insights into hematopoiesis and hematological malignancies through the application of novel single-cell approaches. We particularly focus on biological insights made through the study of stem/progenitors cells in myeloid malignancy at single-cell resolution. Bulk molecular profiling of hematological malignancies by next generation sequencing techniques has provided major insights into the molecular pathogenesis of blood cancers. This technology is now routinely implemented in advanced clinical diagnostics, leading to the development of novel targeted therapies. However, bulk genetic analysis can obscure key aspects of intratumoral heterogeneity which underlies critical disease events, such as treatment resistance and clonal evolution. The past few years have seen an explosion of novel techniques to analyze RNA, DNA, and protein expression at the single-cell level, providing unprecedented insight into cellular heterogeneity. Given the ease of accessibility of liquid tumor biopsies, hematology is well positioned to move novel single-cell techniques towards routine application in the clinic. The present review sets out to discuss current and potential future applications for this technology in the management of patients with hematological cancers.


The Effect of Sandblasting, Er:YAG Laser, and Heat Treatment on the Mechanical Properties of Different Zirconia Cores.
The purpose of this study was to evaluate the effect of surface and heat treatments on the mechanical properties and phase transformation of yttria-stabilized tetragonal zirconia (Y-TZP) materials. Zirconia is exposed to several treatments during dental application process. Knowing the effect of applied treatments on zirconia is essential for clinical success. Forty disk specimens of Zirkonzahn (Z) and DC-Zirkon (DC) materials were fabricated. The specimens were randomly divided into four groups according to surface [control, sandblasting, Erbium: Yttrium Aluminum Garnet (Er:YAG) laser irradiation] and heat (firing) treatments. The surface roughness (Ra, μm) was measured using a surface profilometer. The relative amount of the transformed monoclinic (m) phase was analyzed by X-ray diffractometry (XRD). Biaxial flexural strength was tested using piston-on-three-ball technique. The data were analyzed with the Kruskal-Wallis H test with Bonferroni correction, and the Mann-Whitney U test was used for pairwise comparisons. There were no significant differences in surface roughness among the treated groups (p > 0.05), whereas sandblasting showed higher surface roughness than other treatments for both materials. Scanning electron microscopic (SEM) analyses revealed changes in surface morphology after surface treatments, especially in laser groups with the formation of cracks, and in sandblasting groups with the formation of microretentive grooves. The greatest amount of the monoclinic phase was measured after sandblasting (8.13%) for Z and (19.8%) for DC. The monoclinic phase reverted to the tetragonal phase after heat treatment. Heat treatment groups showed significantly lower flexural strength than other treatments (p < 0.05). Heat and surface treatments influenced the mechanical properties of zirconia ceramic. The biaxial flexural strength and crystalline phase of materials decreased after heat treatments.


Cytokine flow cytometry differentiates the clinical status of multiple sclerosis (MS) patients.
In this study we have examined intracellular cytokines in peripheral blood mononuclear cells (PBMC) of MS patients by flow cytometry (cytokine flow cytometry). MS progressive patients showed an increased number of cells producing interferon-gamma (IFN-gamma) after activation with phorbol 12-myristate 13-acetate and ionomycin, compared with patients with clinically inactive forms (P < 0001) and with healthy controls (P = 0001). These cells belonged to the CD4+ and CD8+ subsets in similar proportions. Clinically inactive patients showed a lower level of cells producing IL-2 than controls (P = 0.03) and active MS patients (P = 0.03). Most IL-2-producing cells were CD4+ lymphocytes, although a small part of the IL-2 was also produced by CD8+ cells. The percentage of cells producing simultaneously IL-2 and IFN-gamma was increased in active MS and they were mainly CD4+ lymphocytes. No differences in the production of IL-4 were observed between groups. However, we found an increased IL-10 production in clinically active MS patients (P = 0.03). Treatment with IFN-beta of active MS patients showed lower levels of cytokines when compared with untreated MS patients. This methodological approach could help in the follow up and therapeutic monitoring of MS patients.


Functional, but not anatomical, separation of "what" and "when" in prefrontal cortex.
How does the brain store information over a short period of time? Typically, the short-term memory of items or values is thought to be stored in the persistent activity of neurons in higher cortical areas. However, the activity of these neurons often varies strongly in time, even if time is unimportant for whether or not rewards are received. To elucidate this interaction of time and memory, we reexamined the activity of neurons in the prefrontal cortex of monkeys performing a working memory task. As often observed in higher cortical areas, different neurons have highly heterogeneous patterns of activity, making interpretation of the data difficult. To overcome these problems, we developed a method that finds a new representation of the data in which heterogeneity is much reduced, and time- and memory-related activities became separate and easily interpretable. This new representation consists of a few fundamental activity components that capture 95% of the firing rate variance of >800 neurons. Surprisingly, the memory-related activity components account for <20% of this firing rate variance. The observed heterogeneity of neural responses results from random combinations of these fundamental components. Based on these components, we constructed a generative linear model of the network activity. The model suggests that the representations of time and memory are maintained by separate mechanisms, even while sharing a common anatomical substrate. Testable predictions of this hypothesis are proposed. We suggest that our method may be applied to data from other tasks in which neural responses are highly heterogeneous across neurons, and dependent on more than one variable.


Evaluation of a rapid antigen test for detection of Streptococcus pneumoniae in cerebrospinal fluid.
Detection of Streptococcus pneumoniae antigen in cerebrospinal fluid (CSF) using lateral flow immunochromatography tests (ICTs) is an effective, rapid and low-cost method to diagnose pneumococcal meningitis. This study evaluated the diagnostic accuracy of the Uni-Gold ICT to detect pneumococcal antigen in CSF specimens, compared with gold standard bacteriology and quantitative real-time PCR (qPCR) testing. CSF specimens (n=69) from patients with suspected bacterial meningitis were included in the study. 13/69 (19%) were positive and 56/69 (81%) were negative for pneumococcus by the gold standard tests. The ICT had sensitivity of 85% (55%-98%), specificity of 96% (88%-100%), positive likelihood ratio of 23.7 (6-94) and negative likelihood ratio of 0.16 (0.04-0.57). Overall, a strong correlation between the ICT and qPCR results was seen (κ=0.81). In contrast, CSF microscopy and culture were exceptionally insensitive. The ICT method is sufficiently robust and accurate for use in algorithms to diagnose bacterial meningitis.


Effects of parylene-C photooxidation on serum-assisted glial and neuronal patterning.
The increasing use of patterned neural networks in multielectrode arrays and similar devices drives the constant development and evaluation of new biomaterials. Recently, we presented a promising technique to guide neurons and glia reliably and effectively. Parylene-C, a common hydrophobic polymer, was photolithographically patterned on silicon oxide (SiO(2)) and subsequently activated via immersion in serum. In this article, we explore the effects of ultraviolet (UV)-induced oxidation on parylene's ability to pattern neurons and glia. We exposed parylene-C stripe patterns to increasing levels of UV radiation and found a dose-dependent reduction in the total mass of patterned cells, as well as a gradual loss of glial and neuronal conformity to the patterns. In contrast, nonirradiated patterns had superior patterning results and increased presence of cells. The reduced cell adhesion and patterning after the formation of aldehyde and carboxyl groups on UV-radiated parylene-C supports our hypothesis that cell adhesion and growth on parylene is facilitated by hydrophobic adsorption of serum proteins. We conclude that unlike other cell patterning schemes, our technique does not rely on photooxidation of the polymer. Nonetheless, the precise control of oxygenated groups on parylene could pave the way for the differential binding of proteins and other molecules on the surface, aiding in the adhesion of alternative cell types. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.


Protein Profile of the Acquired Enamel Pellicle after Rinsing with Whole Milk, Fat-Free Milk, and Water: An in vivo Study.
This study detected changes in the protein profile of the acquired enamel pellicle (AEP) formed in vivo after rinsing with whole milk, fat-free milk, or water. Nine subjects in good oral condition took part in the study. The acquired pellicle was formed in the morning, for 120 min, after prophylaxis with pumice. Following this, the volunteers rinsed with 10 mL of whole milk, fat-free milk, or deionized water for 30 s, following a blinded crossover protocol. After 60 min, the pellicle was collected with filter paper soaked in 3% citric acid and processed for analysis by liquid chromatography-electrospray ionization tandem mass spectrometry. The obtained tandem mass spectrometry spectra were searched against a human protein database (Swiss-Prot). The proteomic data related to protein quantification were analysed using the PLGS software. A total of 260 proteins were successfully identified in the AEP samples collected from all groups. Forty-nine were common to all 3 groups, while 72, 62, and 49 were specific to the groups rinsing with whole milk, fat-free milk, and water, respectively. Some were typical components of the AEP, such as cystatin-B, cystatin-SN, isoforms of α-amylase, IgA and IgG, lysozyme C, protein S100 A78, histatin-1, proline-rich protein 27, statherin, and lactotransferrin. Other proteins are not commonly described as part of the AEP but could act in defence of the organism against pathogens. Distinct proteomic profiles were found in the AEP after rinsing with whole or fat-free milk, which could have an impact on bacterial adhesion and tooth dissolution. The use of fat-free milk could favourably modulate the adhesion of bacteria to the AEP as well as biofilm formation when compared with whole milk.


Using a multi-feature paradigm to measure mismatch responses to minimal sound contrasts in children with cochlear implants and hearing aids.
Our aim was to explore whether a multi-feature paradigm (Optimum-1) for eliciting mismatch negativity (MMN) would objectively capture difficulties in perceiving small sound contrasts in children with hearing impairment (HI) listening through their hearing aids (HAs) and/or cochlear implants (CIs). Children aged 5-7 years with HAs, CIs and children with normal hearing (NH) were tested in a free-field setting using a multi-feature paradigm with deviations in pitch, intensity, gap, duration, and location. There were significant mismatch responses across all subjects that were positive (p-MMR) for the gap and pitch deviants (F(1,43) = 5.17, p = 0.028 and F(1,43) = 6.56, p = 0.014, respectively) and negative (MMN) for the duration deviant (F(1,43) = 4.74, p = 0.035). Only the intensity deviant showed a significant group interaction with MMN in the HA group and p-MMR in the CI group (F(2,43) = 3.40, p = 0.043). The p-MMR correlated negatively with age, with the strongest correlation in the NH subjects. In the CI group, the late discriminative negativity (LDN) was replaced by a late positivity with a significant group interaction for the location deviant. Children with severe HI can be assessed through their hearing device with a fast multi-feature paradigm. For further studies a multi-feature paradigm including more complex speech sounds may better capture variation in auditory processing in these children.


Human Capital Acquisition and Occupational Choice: Implications for Economic Development.
Using household-level data from Mexico we document patterns among schooling, entrepreneurial decisions and household characteristics such as assets, talent of household members and age of the household head. Motivated by our findings, we develop a heterogeneous-agent, incomplete-markets, overlapping-generations dynasty model. Households jointly decide over their life cycle on (i) kids' human capital investments (schooling) and (ii) parents' entry, exit and investment into alternative entrepreneurial modes (subsistence and modern). With financial constraints all of these are co-determined. A calibrated version of our model can account for the broad correlation patterns uncovered in the data within and across generations, e.g., a non-monotonic relationship between educational choices and assets across occupations, growth in profits and employment for modern firms only, and dynastic persistence across generations in education and wealth. Endogenous human capital acquisition is a key driver of inequality and intergenerational persistence. Eliminating this channel would decrease the top 10% income share by 47%. Eliminating within-period borrowing constraints would increase average household expenditure by 7.1% and benefit the middle class, reducing top and bottom expenditure shares. It would also reduce by 28% the correlation between household assets and kids' schooling levels.


Brassinosteroid signalling.
The brassinosteroid (BR) class of steroid hormones regulates plant development and physiology. The BR signal is transduced by a receptor kinase-mediated signal transduction pathway, which is distinct from animal steroid signalling systems. Recent studies have fully connected the BR signal transduction chain and have identified thousands of BR target genes, linking BR signalling to numerous cellular processes. Molecular links between BR and several other signalling pathways have also been identified. Here, we provide an overview of the highly integrated BR signalling network and explain how this steroid hormone functions as a master regulator of plant growth, development and metabolism.


Transforming growth factor-beta inhibits the expression of clock genes.
Disturbances of sleep-wake rhythms are an important problem in Alzheimer's disease (AD). Circadian rhythms are regulated by clock genes. Transforming growth factor-beta (TGF-β) is overexpressed in neurons in AD and is the only cytokine that is increased in cerebrospinal fluid (CSF). Our data show that TGF-β2 inhibits the expression of the clock genes Period (Per)1, Per2, and Rev-erbα, and of the clock-controlled genes D-site albumin promoter binding protein (Dbp) and thyrotroph embryonic factor (Tef). However, our results showed that TGF-β2 did not alter the expression of brain and muscle Arnt-like protein-1 (Bmal1). The concentrations of TGF-β2 in the CSF of 2 of 16 AD patients and of 1 of 7 patients with mild cognitive impairment were in the dose range required to suppress the expression of clock genes. TGF-β2-induced dysregulation of clock genes may alter neuronal pathways, which may be causally related to abnormal sleep-wake rhythms in AD patients.


Synergistic inhibition of PARP-1 and NF-κB signaling downregulates immune response against recombinant AAV2 vectors during hepatic gene therapy.
Host immune response remains a key obstacle to widespread application of adeno-associated virus (AAV) based gene therapy. Thus, targeted inhibition of the signaling pathways that trigger such immune responses will be beneficial. Previous studies have reported that DNA damage response proteins such as poly(ADP-ribose) polymerase-1 (PARP-1) negatively affect the integration of AAV in the host genome. However, the role of PARP-1 in regulating AAV transduction and the immune response against these vectors has not been elucidated. In this study, we demonstrate that repression of PARP-1 improves the transduction of single-stranded AAV vectors both in vitro (∼174%) and in vivo (two- to 3.4-fold). Inhibition of PARP-1, also significantly downregulated the expression of several proinflammatory and cytokine markers such as TLRs, ILs, NF-κB subunit proteins associated with the host innate response against self-complementary AAV2 vectors. The suppression of the inflammatory response targeted against these vectors was more effective upon combined inhibition of PARP-1 and NF-κB signaling. This strategy also effectively attenuated the AAV capsid-specific cytotoxic T-cell response, with minimal effect on vector transduction, as demonstrated in normal C57BL/6 and hemophilia B mice. These data suggest that targeting specific host cellular proteins could be useful to attenuate the immune barriers to AAV-mediated gene therapy.


The Cinchona Program (1940-1945): science and imperialism in the exploitation of a medicinal plant.
During World War II, the United States implemented programs to exploit hundreds of raw materials in Latin America, many of them botanical. This required the participation of the country's scientific community and marked the beginning of intervention in Latin American countries characterized by the active participation of the United States in negotiations (and not only by private firms supported by the United States). Many federal institutions and companies were created, others were adapted, and universities, research centers and pharmaceutical companies were contracted. The programs undertaken by this coalition of institutions served to build and consolidate the dependence of Latin American countries on United States technology, to focus their economies on the extraction and development of resources that the United States could not obtain at home (known as "complementary") and to impede the development of competition. Latin American republics had been historically dependant on raw material exports (minerals and plants). But during World War II their dependence on U.S. loans, markets, science and technology reached record levels. One example of this can be appreciated through a careful examination of the Cinchona Program, implemented in the 1940s by US agencies in Latin America. This program for the extraction of a single medicinal plant, apart from representing a new model of scientific imperialism (subsequently renamed "scientific cooperation") was the most intensive and extensive scientific exploration of a single medicinal plant in the history of mankind.


Pretransplant serum troponin levels are highly predictive of patient and graft survival following liver transplantation.
Optimizing the utility of liver transplantation requires the identification of factors that confer increased risk of posttransplant mortality. Elevated serum troponin (TN) levels are strongly predictive of posttransplant mortality after kidney transplantation. We sought to determine whether pretransplant TN levels were predictive of mortality and graft loss after liver transplantation in 236 liver transplant recipients from 1998 to 2001 with 8.2 years of follow-up. Elevated TN levels [hazard ratio (HR) = 2.19, P = 0.004] and a pretransplant history of cardiovascular disease (CVD; HR = 1.90, P = 0.031) were predictive of patient mortality. Elevated TN levels (HR = 2.44, P < 0.001), a history of CVD (HR = 1.83, P = 0.031), and a combination of elevated TN levels and CVD (HR = 2.75, P = 0.027) were associated with increased graft loss. Multivariate analysis confirmed TN and CVD as independent predictors of mortality and graft loss. CVD (HR = 2.39, P = 0.032) and a combination of elevated TN levels and a history of CVD (HR = 6.67, P < 0.001) were predictive of graft loss within 1 year. Age, smoking, diabetes, hypertension, obesity, creatinine levels, and Model for End-Stage Liver Disease scores were not predictive of posttransplant mortality or graft loss. In summary, elevated pretransplant serum TN levels are strongly predictive of mortality and graft loss after liver transplantation and may be helpful in risk stratification of potential liver transplant recipients.


Regulation of tumor necrosis factor alpha gene expression by mycobacteria involves the assembly of a unique enhanceosome dependent on the coactivator proteins CBP/p300.
Tumor necrosis factor alpha (TNF-alpha) plays an important role in host containment of infection by Mycobacterium tuberculosis, one of the leading causes of death by an infectious agent globally. Using the pathogenic M. tuberculosis strain H37Rv, we present evidence that upon stimulation of monocytic cells by M. tuberculosis a unique TNF-alpha enhanceosome is formed, and it is distinct from the TNF-alpha enhanceosome that forms in T cells stimulated by antigen engagement or virus infection. A distinct set of activators including ATF-2, c-jun, Ets, Sp1, Egr-1 and the coactivator proteins CBP/p300 are recruited to the TNF-alpha promoter after stimulation with M. tuberculosis. Furthermore, the formation of this enhanceosome is dependent on inducer-specific helical phasing relationships between transcription factor binding sites. We also show that the transcriptional activity of CBP/p300 is potentiated by mycobacterial stimulation of monocytes. The identification of TNF-alpha regulatory elements and coactivators involved in M. tuberculosis-stimulated gene expression thus provides potential selective molecular targets in the modulation of TNF-alpha gene expression in the setting of mycobacterial infection.


Functional domains in the bacteriophage phi 29 terminal protein for interaction with the phi 29 DNA polymerase and with DNA.
Deletion mutants at the amino- and carboxyl-ends of the phi 29 terminal protein, as well as internal deletion and substitution mutants, whose ability to prime the initiation of phi 29 DNA replication was affected to different extent, have been assayed for their capacity to interact with DNA or with the phi 29 DNA polymerase. One DNA binding domain at the amino end of the terminal protein has been mapped. Two regions involved in the binding to the DNA polymerase, an internal region near the amino-terminus and a carboxyl-terminal one, have been also identified. Interaction with both DNA and phi 29 DNA polymerase are required to led to the formation of terminal protein-dAMP initiation complex to start phi 29 DNA replication.


Study of parameters in focus simulation functions of virtual slide.
As a special function of Virtual Slide (VS) for thick specimens like cytology slides, multilayer (Z-stack) simulated focus and focus fusion were introduced. From the standpoint of surgical pathologist, the optimum parameters for multilayer focus simulation were examined. First, minimal thickness of the layer was checked by measuring thickness of small cells counting the number of the layers that come into focus. Then the optimal number of layers to scan, total thickness, was tried. Small-sized cell nuclei showed around 2 μm or less thickness. As minimal thickness of one layer for focus simulation, less than 2 μm is required. Papillary cell mass of urothelial carcinoma, aspiration cytology specimen of breast or thyroid, and uterine cervical smear showed different optimal thickness. Cells piling up more than 4 to 5 layer are difficult to make close up observation. Total 15 (to 30) μm thick scan was enough for most specimens. The "focus fusion" image is single layer image synthesized from multiple layer images. Several layer thicknesses were examined, and there was negligible difference between the focus fusion image synthesized from 0.25 and 1 μm thick layers. In the focus fusion image synthesized from 3 μm thick layers, some cells not to come into focus. The "focus fusion" seems to contain all the cells in one plane, and easy for screening. To emphasize the existence of myoepithelial cells in fibroadenoma of breast, or to clarify the 3-dimensional tissue structure, multilayer image was better. From our results, 10 layers with 1.5 μm thick each provide sufficient information in most specimens.


Intravenous and intracoronary fibrinolytic therapy in acute myocardial infarction: overview of results on mortality, reinfarction and side-effects from 33 randomized controlled trials.
During the past 25 years, 24 randomized trials of intravenous (IV) fibrinolytic treatment have been reported, involving a total of some 6000 patients in the acute phase of myocardial infarction. Most tested IV streptokinase (SK), but a few tested IV urokinase (UK). In the past 2 or 3 years numerous small randomized trials of intracoronary (IC) SK have been started, 9 of which, involving a total of about 1000 such patients, have been reported. Because all of these IV and IC trials were small (the largest including only 747 patients), their separate results appear contradictory and unreliable. But, an overview of the data from these trials indicates that IV treatment produces a highly significant (22% +/- 5%, P less than 0.001) reduction in the odds of death, an even larger reduction in the odds of reinfarction, and an absolute frequency of serious adverse effects to set against this that is much smaller than the absolute mortality reduction. The apparent size of the mortality reduction in the IV trials was similar whether anticoagulants were compulsory or optional, whether treatment was in a coronary care unit or an ordinary ward and, surprisingly, whether treatment began early (less than 6 h from onset of symptoms) or late (generally 12-24 h). In addition, there was no evidence that UK was more effective than the less expensive SK, or that, despite their technical complexity, the new IC regimes were more effective than the old IV regimes. Even the IV schedules that have been studied in randomized trials were, however, quite complex, and the IC schedules were far more so. Perhaps partly because of this, none of them is widely used. If so, then some much simpler, and hence more widely practicable, IV SK regimes should be developed and tested. For example, a simple one hour high-dose IV SK infusion, without anticoagulation, will successfully convert virtually all of the available plasminogen into plasmin. But, it may be several years before the net effects on mortality of any more widely practicable IV SK regimes can be agreed unless many of the hospitals that do not wish routinely to use IC regimes or the complex previous IV regimes will collaborate in multicentre randomized trials that can, if necessary, continue rapid intake until some tens of thousands of patients have been randomized, and some thousands of deaths have been observed among the control and treated patients. The same, of course, may be true for any other fibrinolytic regimes (e.g. infusion of tissue plasminogen activator) if their net effects on mortality are comparable to those of IV SK.


Identification of proteins that may directly interact with human RPA.
RPA, which consisted of three subunits (RPA1, 2 and 3), plays essential roles in DNA transactions. At the DNA replication forks, RPA binds to single-stranded DNA region to stabilize the structure and to assemble other replication proteins. Interactions between RPA and several replication proteins have been reported but the analysis is not comprehensive. We systematically performed the qualitative analysis to identify RPA interaction partners to understand the protein-protein interaction at the replication forks. We expressed in insect cells the three subunits of human RPA, together with one replication protein, which is present at the forks under normal conditions and/or under the replication stress conditions, to examine the interaction. Among 30 proteins examined in total, it was found that at least 14 proteins interacted with RPA. RPA interacted with MCM3-7, MCM-BP and CDC45 proteins among the proteins that play roles in the initiation and the elongation of the DNA replication. RPA bound with TIPIN, CLASPIN and RAD17, which are involved in the DNA replication checkpoint functions. RPA also bound with cyclin-dependent kinases and an amino-terminal fragment of Rb protein that negatively regulates DNA replication. These results suggest that RPA interacts with the specific proteins among those that play roles in the regulation of the replication fork progression.


An approach to management of critical indoor air problems in school buildings.
This study was conducted in a school center that had been the focus of intense public concern over 2 years because of suspected mold and health problems. Because several attempts to find solutions to the problem within the community were not satisfactory, outside specialists were needed for support in solving the problem. The study group consisted of experts in civil engineering, indoor mycology, and epidemiology. The studies were conducted in close cooperation with the city administration. Structures at risk were opened, moisture and temperature were measured, and the causes of damage were analyzed. Microbial samples were taken from the air, surfaces, and materials. Health questionnaires were sent to the schoolchildren and personnel. Information on the measurements and their results was released regularly to school employees, students and their parents, and to the media. Repairs were designed on the basis of this information. Moisture damage was caused mainly by difficult moisture conditions at the building site, poor ventilation, and water leaks. Fungal genera (concentrations <200 colony-forming units (cfu)/m(3), <3000 cfu/cm(2)) typical to buildings with mold problems (e.g., Aspergillus versicolor, Eurotium) were collected from the indoor air and surfaces of the school buildings. Where moisture-prone structures were identified and visible signs of damage or elevated moisture content were recorded, the numbers of microbes also were high; thus microbial results from material samples supported the conclusions made in the structural studies. Several irritative and recurrent symptoms were common among the upper secondary and high school students. The prevalence of asthma was high (13%) among the upper secondary school students. During the last 4 years, the incidence of asthma was 3-fold that of the previous 4-year period.


Medical students' perceptions of resident teaching: have duty hours regulations had an impact?
This study describes medical students' perceptions about resident teaching on a surgery clerkship and examines student perceptions before and after the implementation of duty hours regulations (DHR). There has been much discussion about the impact of DHR on surgical education. One area that merits evaluation is the effect that DHR have had on student education. Learners perceive the clinical teacher role as comprised of 4 roles: teacher, person, physician, and supervisor. This model served as the basis for examining resident teaching before and after DHR. Students completed end-of-rotation evaluations about residents' teaching effectiveness, amount of feedback, and quality of interactions. Student comments were compiled into individual resident reports, and reports were collected from pre- (2002-2003) and post- (2003-2004) DHR. A coding scheme was developed to describe resident performance in 4 roles: teacher, person, physician, and supervisor. Three coders independently reviewed 124 resident reports maintaining an interrater agreement of 80%. Analyses of variance were conducted to compare data from pre- and post-DHR. After implementation of DHR, there were significantly more negative comments (P = 0.005), including comments about residents as supervisor (P = 0.001), teacher (P = 0.027), and teaching activities (P = 0.001). Positive comments about bedside teaching decreased (P = 0.007). Although total positive comments about resident as person increased (P = 0.01), total negative comments about resident as person also increased (P = 0.02). Findings of this study indicate that DHR have had a negative impact on medical students' perceptions of resident teaching. Surgical educators must develop programs that address resident teaching skills in a different environment.


Rattling the plate--reasons and rationales for early weaning.
To identify a range of attitudes and beliefs which influence the timing of introduction to solid food, five focus group discussions were undertaken within a maternity hospital setting. These sessions explored early feeding behaviour, stimuli to changing feeding habits and subsequent responses in 22 primiparous and seven multiparous mothers (mean age 27.0+/-4.8 years) with babies aged 8-18 weeks (mean age 13.0+/-4.2 weeks). One-third of the participants had introduced solid food to their infants (mean age of introduction 11.6 weeks, range 2-16 weeks). Mothers believed that the introduction of solids was baby led and initiated by some physical characteristic or behavioural action of the infant. All mothers were aware of current recommendations to avoid the introduction of solid food until 4 months. Few knew why this should be and concepts of long-term ill health were difficult to conceptualize. The conflict between rigid feeding guidelines and flexible advice from supportive health professionals created confusion over the importance of good weaning practices. The current findings highlight issues relevant to the introduction of solid food, and provide a foundation for further research which can identify the relative importance of these factors and provide a rationale for the design of contemporary intervention strategies.


Rapid identification viruses from nasal pharyngeal aspirates in acute viral respiratory infections by RT-PCR and electrospray ionization mass spectrometry.
Diagnosis of the etiologic agent of respiratory viral infection relies traditionally on culture or antigen detection. This pilot evaluation compared performance characteristics of the RT-PCR and electrospray ionization mass spectrometry (RT-PCR/ESI-MS) platform to conventional virologic methods for identifying multiple clinically relevant respiratory viruses in nasopharyngeal aspirates. The RT-PCR/ESI-MS respiratory virus surveillance kit was designed to detect respiratory syncytial virus, influenza A and B, parainfluenza types 1-4, adenoviridae types A-F, coronaviridae, human bocavirus, and human metapneumovirus. Patients (N=192) attending an emergency department during the 2007-2008 respiratory season consented, and "excess" frozen archived nasopharyngeal aspirates were analysed; 46 were positive by conventional virology and 69 by RT-PCR/ESI-MS, among which there were six samples with multiple viral pathogens detected. The sensitivity and specificity of the assay were 89.1% and 80.3%, respectively. Additional viruses that were not identified by conventional virology assays were detected (4 human bocaviruses and 7 coronaviruses). Samples in which the RT-PCR/ESI-MS results disagreed with conventional virology were sent for analysis by a third method using a commercial RT-PCR-based assay, which can identify viruses not detectable by conventional virologic procedures. Time to first result of RT-PCR/ESI-MS was 8h. RT-PCR/ESI-MS demonstrated capacity to detect respiratory viruses identifiable and unidentifiable by conventional methods rapidly.


Comprehensive panel of real-time TaqMan polymerase chain reaction assays for detection and absolute quantification of filoviruses, arenaviruses, and New World hantaviruses.
Viral hemorrhagic fever is caused by a diverse group of single-stranded, negative-sense or positive-sense RNA viruses belonging to the families Filoviridae (Ebola and Marburg), Arenaviridae (Lassa, Junin, Machupo, Sabia, and Guanarito), and Bunyaviridae (hantavirus). Disease characteristics in these families mark each with the potential to be used as a biological threat agent. Because other diseases have similar clinical symptoms, specific laboratory diagnostic tests are necessary to provide the differential diagnosis during outbreaks and for instituting acceptable quarantine procedures. We designed 48 TaqMan-based polymerase chain reaction (PCR) assays for specific and absolute quantitative detection of multiple hemorrhagic fever viruses. Forty-six assays were determined to be virus-specific, and two were designated as pan assays for Marburg virus. The limit of detection for the assays ranged from 10 to 0.001 plaque-forming units (PFU)/PCR. Although these real-time hemorrhagic fever virus assays are qualitative (presence of target), they are also quantitative (measure a single DNA/RNA target sequence in an unknown sample and express the final results as an absolute value (e.g., viral load, PFUs, or copies/mL) on the basis of concentration of standard samples and can be used in viral load, vaccine, and antiviral drug studies.


Different roles of membrane potentials in electrotaxis and chemotaxis of dictyostelium cells.
Many types of cells migrate directionally in direct current (DC) electric fields (EFs), a phenomenon termed galvanotaxis or electrotaxis. The directional sensing mechanisms responsible for this response to EFs, however, remain unknown. Exposing cells to an EF causes changes in plasma membrane potentials (V(m)). Exploiting the ability of Dictyostelium cells to tolerate drastic V(m) changes, we investigated the role of V(m) in electrotaxis and, in parallel, in chemotaxis. We used three independent factors to control V(m): extracellular pH, extracellular [K(+)], and electroporation. Changes in V(m) were monitored with microelectrode recording techniques. Depolarized V(m) was observed under acidic (pH 5.0) and alkaline (pH 9.0) conditions as well as under higher extracellular [K(+)] conditions. Electroporation permeabilized the cell membrane and significantly reduced the V(m), which gradually recovered over 40 min. We then recorded the electrotactic behaviors of Dictyostelium cells with a defined V(m) using these three techniques. The directionality (directedness of electrotaxis) was quantified and compared to that of chemotaxis (chemotactic index). We found that a reduced V(m) significantly impaired electrotaxis without significantly affecting random motility or chemotaxis. We conclude that extracellular pH, [K(+)], and electroporation all significantly affected electrotaxis, which appeared to be mediated by the changes in V(m). The initial directional sensing mechanisms for electrotaxis therefore differ from those of chemotaxis and may be mediated by changes in resting V(m).


Evaluation of risk of liver metastasis in colorectal adenocarcinoma based on the combination of risk factors including CD10 expression: multivariate analysis of clinicopathological and immunohistochemical factors.
Evaluation of the relationship between clinicopathological and immunohistochemical risk factors for liver metastasis, including CD10 expression, is meaningful in colorectal carcinoma (CRC). The purpose of the present study was to clarify what kind of risk factors are significant and independent factors for the development of liver metastasis in CRC. Sixty cases of advanced CRC with synchronous liver metastasis and sixty cases of advanced CRC without liver metastasis at least 5 years after resection of the primary CRC were randomly selected. We analysed the clinicopathological factors and the expression of four biological factors, CD44, transforming growth factor alpha (TGF-alpha), vascular endothelial growth factor (VEGF) and CD10, by immunohistochemistry. Univariate analysis revealed that the incidence of vascular invasion, the incidence of lymph node metastasis and the expression of CD44, TGF-alpha, VEGF and CD10 were all significantly higher in the cases of CRC with liver metastasis than in cases of non-metastatic CRC. Logistic regression analysis showed that lymph node metastasis, the expression of CD10 and the expression of VEGF were significant factors and independent of the other variables. If all three markers are positive, the probability of liver metastasis becomes 92.7%. In this study, lymph node metastasis, CD10 and VEGF were all found to be significant risk factors for the development of liver metastasis in the cases of CRC. These risk factors according to multivariate analysis are candidate markers for predicting the development of liver metastasis.


Doxepin for Radiation Therapy-Induced Mucositis Pain in the Treatment of Oral Cancers.
Radiotherapy (RT), an integral part of the oncologic treatment for patients with head and neck cancer, can cause adverse side effects such as oral mucositis (OM). Pain from OM can impact a patient's quality of life and interrupt RT treatment schedules, which decreases the probability for achieving cancer cure. Conventionally, RT-induced OM pain is treated with analgesics and/or mouthwash rinses. Doxepin, a traditional tricyclic antidepressant with analgesic and anesthetic properties when applied topically to the mucosa, has been shown to lower OM pain in multiple single-arm trials (Epstein et al.) and more recently, in a placebo-controlled crossover study (Leenstra and Miller et al.). Currently, a placebo-controlled study (Sio and Miller et al.) using doxepin for esophagitis pain caused by RT to the thorax is underway. Doxepin will also be further compared with magic mouthwash and a placebo solution in a three-arm trial (Miller and Sio et al.) with head and neck cancer patients with OM pain caused by RT. Doxepin may represent a new standard for treating RT-induced OM pain in the future.


Ultralight supercapacitors utilizing waste cotton pads for wearable energy storage.
Successful popularization of wearable energy storage devices lies in the exploitation of scalable fabrication technologies that are based on economically viable materials. Herein, we reveal that discarded cotton pads can be used as a cost-efficient substrate for the in situ polymerization of pyrrole and exhibited good mechanical flexibility, lightness, and high conductivity. To extend the applications of the resulting PPy-coated cotton pads (PCPs) to the supercapacitor field, a layer of MnO2 nanosheets was further decorated on the surface of PCPs (PCPs@MnO2) by a simple electrochemical deposition technique. The PPy coating not only improves the electrical conductivity of the cotton pads, but also increases the contact between the active materials and the cotton fibers. Amazingly, ultrathin (≈ 0.8 mm) flexible solid-state asymmetric supercapacitors (ASCs) using PCPs@MnO2 as the positive electrode and active carbon coated on PCPs (PCPs@AC) as the negative electrode display a high areal capacitance of 1.21 F cm-2 at 1 mA cm-2, and a high energy density of 6.8 mW h cm-3 at a power density of 11.2 mW cm-3, which can also be tailored and folded into various shapes with only slight capacitance fading. These findings demonstrate that the prepared advanced ultralight, flexible and renewable cotton pads hold great promise for practical application in wearable energy storage systems with high cost effectiveness and scalability.


Dynamics of the conformational transitions during the dimerization of an intrinsically disordered peptide: a case study on the human islet amyloid polypeptide fragment.
Amyloid deposits of human islet amyloid polypeptide (hIAPP) are identified in 95% of type II diabetes patients. The oligomers during the early stage of hIAPP aggregation are believed to be more cytotoxic than the mature fibrils. However, the structural details during the initial stage of hIAPP aggregation are still under debate experimentally. To understand its initial nucleation mechanism, we investigate the thermodynamics and kinetics of hIAPP(11-25) dimerization, which is the first manifestation of the interplay between intra- and inter-molecular interactions, via the construction of Markov state models from extensive molecular dynamics simulations. We identified a largely populated metastable dimer state with the antiparallel cross-β structure, although tangled coil states are also observed. The dimerization process consists of two stages kinetically: the initial collision of separate monomers followed by structural rearrangements. During the collapsing stage, hydrophobic interactions are the main driving force, although electrostatic interactions also play a role. In the subsequent structural rearrangement step, there exist heterogeneous pathways from the initial collapsed complexes to the antiparallel cross-β structure, with the transition time-scales around hundreds of microseconds. Our replica-exchange molecular dynamics simulations demonstrate that this antiparallel cross-β state is negligible in the dimer ensemble of the fibril-free S20P mutant, indicating that it is an on-pathway intermediate for hIAPP(11-25) fibrillation. These results, together with those from our previous study of the monomer, prompt us to propose a generalized model with the combination of the induced-fit and conformational-selection mechanisms for this dimerization process. These findings shed light on the understanding of hIAPP(11-25) aggregation mechanisms.


Cold acclimation-induced up-regulation of the ribosomal protein L7 gene in the freeze tolerant wood frog, Rana sylvatica.
Natural freezing survival by the wood frog, Rana sylvatica, involves multiple organ-specific changes in gene expression. The present study used differential display PCR to find cold-responsive genes in wood frog skin. A cDNA was retrieved from skin that was in higher amounts in cold- versus warm-acclimated frogs. The cDNA was used to probe a wood frog liver cDNA library and retrieve a long sequence that, after the further application of 5'RACE, was shown to encode the full sequence of the ribosomal large subunit protein 7 (RPL7) (GenBank accession number AF175983). Wood frog RPL7 contained 246 amino acids and shared 90% identity with Xenopus laevis RPL7, 82-83% with chicken and zebrafish homologues, and 79% with mammalian RPL7. Multiple binding domains found in human RPL7 showed differing degrees of conservation in the frog protein. Transcript levels of rpl7 were elevated up to 4-fold in skin of cold-acclimated frogs as compared with warm-acclimated animals. Organ-specific responses by rpl7 transcripts also occurred when frogs were given survivable freezing exposures. Transcripts rose by 1.8-3.3 fold in brain and skeletal muscle during freezing but were unaffected in central organs such as liver and heart. Up-regulation of rpl7 also occurred in brain of anoxia-exposed frogs and RPL7 protein levels increased strongly in heart under both freezing and dehydration stresses. Cold- and freezing-responsive up-regulation of the rpl7 gene and RPL7 protein in selected organs suggests that targeted changes in selected ribosomal proteins may be an integral part of natural freeze tolerance.


Spacer devices for inhaled therapy: why use them, and how?
We present an extensive review of the literature to date pertaining to the rationale for using a spacer/valved holding chamber (VHC) to deliver inhaled therapy from a pressurised, metered-dose inhaler, a discussion of how the properties of individual devices may vary according to their physical characteristics and materials of manufacture, the potential risks and benefits of ancillaries such as valves, and the evidence that they contribute tangibly to the delivery of therapy. We also reiterate practical recommendations for the correct usage and maintenance of spacers/VHCs, which we trust offer practical help and advice to patients and healthcare professionals alike.


Hierarchy of ventricular pacemakers.
To characterize the pattern of pacemaker dominance in the ventricular specialized conduction system (VSCS), escape ventricular pacemakers were localized and quantified in vivo and in virto, in normal hearts and in hearts 24 hours after myocardial infarction. Excape pacemaker foci were localized in vivo during vagally induced atrial arrest by means of electrograms recorded from the His bundle and proximal bundle branches and standard electrocardiographic limb leads. The VSCS was isolated using a modified Elizari preparation or preparations of each bundle branch. Peacemakers were located by extra- and intracellular recordings. Escape pacemaker foci in vivo were always in the proximal conduction system, usually the left bundle branch. The rate was 43+/-11 (mean+/-SD) beats/min. After beta-adrenergic blockade, the mean rate fell to 31+/-10 beats/min, but there were no shifts in pacemaker location. In the infarcted hearts, pacemakers were located in the peripheral left bundle branch. The mean rate was 146+/-20 beats/min. In isolated normal preparations, the dominant pacemakers usually were in the His bundle, firing at a mean rate of 43+/-10 beats/min. The rates of pacemakers diminished with distal progression. In infarcted hearts, the pacemakers invariably were in the infarct zone. The mean firing rates were not influenced by beta-adrenergic blockade. The results indicate that the dominant pacemakers are normally in the very proximal VSCS, but after myocardial infarction pacemaker dominance is shifted into the infarct. Distribution of pacemaker dominance is independent of sympathetic influence.


Advanced Methods for the Investigation of Cell Contact Dynamics in Endothelial Cells Using Florescence-Based Live Cell Imaging.
Endothelial cells of the vascular system are dynamic cells whose molecular adaptability is decisive for the adjustment of homeostasis and organ perfusion. Advanced microscopic techniques, automation processing, and image analysis software was shown to improve the understanding of vascular biology. In this work, we describe advanced methods that allow investigating the dynamics of endothelial cell contacts. The development of viral vectors has contributed significantly to the genetic manipulation of endothelial cells. We used the Gibson assembly as a quick and cheap cloning system for introducing sequences into the lentiviral-based pFUGW vector. Furthermore, classical fluorescence tags such as mCherry and EGFP were compared with self-labeling tags such as Halo and SNAP for their suitability to study junction dynamics in cultured endothelium, and found the self-labeling tags as useful tools. Using such combinations, we found maintained cell junction integrity during shear stress-induced junction remodeling using VE-cadherin-EGFP. Remodeling was accompanied by VE-cadherin plaque formation, indicating that this process is mediated by the for-mation of the actin-driven junction-associated intermittent lamellipodia, JAIL. The combined methods including the Gibson assembly, lentiviral mediated gene transfer, spinning disk-based live cell imaging, and software for quantification allow analyses of the endothelial cell junction dynamics under static and under shear stress conditions.


Videofluoroscopic assessment of muscle fiber shortening in the in situ canine diaphragm.
We tested the hypothesis that dynamic shortening of the costal diaphragm can be accurately estimated from measurements of the radiographic width of the zone of apposition (WZapp) by studying seven supine anesthetized dogs. Both muscle fiber length, represented by the distance between implanted radiopaque markers, and WZapp were measured from digitized recordings of fluoroscopic images utilizing interactive computer software. The WZapp was highly correlated with the length of costal fibers during active respiration in all animals (mean R2 = 0.94). The accuracy in the prediction of fiber length and shortening during breathing is enhanced by inclusion of additional variables describing the displacement of the abdominal wall and the resting geometric orientation of the fibers. We conclude that dynamic fluoroscopic measurement of WZapp is a valuable technique for estimating dynamic diaphragm fiber length and shortening. Depending on the experimental circumstances, WZapp may be a more easily acquired indicator of diaphragm shortening than other variables that have been previously utilized. As such, it may provide a suitable approach to assess active shortening of the diaphragm in humans.


Trends in Dengue Cases Imported into the United States from Pan America 2001-2012.
The objective of this study was to improve risk assessments of travel on dengue (DEN) virus (DENV) distribution. We investigated the exposure risk of US citizens traveling to DEN-endemic Pan American countries. The number of DEN cases reported in 51 Pan American countries from 2001 to 2012 was compared to the population of the same countries. The number of US travelers visiting the Pan American countries was categorized by region, and travel-related DEN infections were analyzed. US residents visiting the Dominican Republic exhibited the highest traveler-related DEN incidence. Brazil showed the most DEN cases in its residents (>1 million reported cases in 2010). The number of DEN cases continues to rise as does international travel and the geographic range of potential DENV vectors. DENV risk assessments may be improved by analyzing the possible routes of entry. Underreporting remains an issue for calculating DENV transmission risk by country and region.


Teaching medical students to express empathy by exploring patient emotions and experiences in standardized medical encounters.
To increase medical students' ability to detect contextual and emotional cues and to respond empathetically to patients. a training course in communication skills and patient-centered care with different teaching activities (didactic, reflective and interactive: workshops and encounters with simulated patients) was delivered to third-year medical students just before their clerkships. The program was evaluated by an external observer (OE) and simulated patients (SP) in 2 or 3 videotaped encounters. Students improved significantly from baseline to 3rd interview in all communicative skills and domains explored both in OE (32.4%) and SP (38.3%) measurement. At the end of the course students detected significantly more clues and made more empathetic expressions. The course seems to improve the ability of students to explore the illness experience, showing more empathy in a more genuine way. This was carried out in consultations lasting 10min. The program is effective and feasible to be applied as a regular formative activity. Further research is needed to assess whether this training program is applicable to students in more advanced educational levels and if it has any additional outcomes.


The birth weight lowering C-allele of rs900400 near LEKR1 and CCNL1 associates with elevated insulin release following an oral glucose challenge.
The first genome-wide association study on birth weight was recently published and the most significant associated birth weight lowering variant was the rs900400 C-allele located near LEKR1 and CCNL1. We aimed to replicate the association with birth weight in the Danish Inter99 study and furthermore to evaluate associations between rs900400 and indices of insulin secretion and insulin sensitivity obtained by oral glucose tolerance tests in adults from the Danish Inter99 study and the Finnish, Metabolic Syndrome in Men (METSIM) sample. For 4,744 of 6,784 Inter99 participants, midwife journals were traced through the Danish State Archives and association of rs900400 with birth weight was examined. Associations between rs900400 and fasting serum insulin, fasting plasma glucose, insulinogenic index, homeostasis model assessment of insulin resistance (HOMA-IR) and disposition index were studied in 5,484 Danish and 6,915 Finnish non-diabetic individuals and combined in meta-analyses. The C-allele of rs900400 was associated with a 22.1 g lower birth weight ([-41.3;-3.0], P = 0.024) per allele. Moreover, in combined analyses of the Danish Inter99 study and the Finnish METSIM study we found that the birth weight lowering allele was associated with increased insulin release measured by the insulinogenic index (β = 2.25% [0.59; 3.91], P = 0.008) and with an increased disposition index (β = 1.76% [0.04; 3.49], P = 0.05). The birth weight lowering effect of the C-allele of rs900400 located near LEKR1 and CCNL1 was replicated in the Danish population. Furthermore the C-allele was associated with increased insulin response following oral glucose stimulation in a meta-analysis based on Danish and Finnish non-diabetic individuals.


Glucose in the hypothalamic paraventricular nucleus regulates GLP-1 release.
Glucokinase (GK) is highly expressed in the hypothalamic paraventricular nucleus (PVN); however, its role is currently unknown. We found that GK in the PVN acts as part of a glucose-sensing mechanism within the PVN that regulates glucose homeostasis by controlling glucagon-like peptide 1 (GLP-1) release. GLP-1 is released from enteroendocrine L cells in response to oral glucose. Here we identify a brain mechanism critical to the release of GLP-1 in response to oral glucose. We show that increasing expression of GK or injection of glucose into the PVN increases GLP-1 release in response to oral glucose. On the contrary, decreasing expression of GK or injection of nonmetabolizable glucose into the PVN prevents GLP-1 release. Our results demonstrate that gluco-sensitive GK neurons in the PVN are critical to the response to oral glucose and subsequent release of GLP-1.


Predicting pediatric length of stay and acuity of care in the first ten minutes with artificial neural networks.
OBJECTIVE: To evaluate the efficacy of artificial neural networks in categorizing pediatric trauma patients into four distinct acuity of care groups and in determining the length of stay (LOS) within specific areas of the hospital. DESIGN: Using historical information from >8,000 pediatric trauma patient records, train and evaluate artificial neural networks to predict the injury severity and LOS for each patient in pediatric intensive care units (PICUs), step-down units, and floor units. Each artificial neural network is evaluated for categorization accuracy and mean absolute error difference on the predicted LOS. SUBJECTS: A total of 10,353 patient records from the National Pediatric Trauma registry, representing all pediatric trauma patients treated at affiliated hospitals from April 1994 through December 1996. Records with incomplete information were eliminated from the study, leaving 8,081 usable patient records. MEASUREMENTS: A total of 14 variables are selected from the 81 values present in the National Pediatric Trauma Registry as independent variables for the artificial neural networks. Each neural network produces nine output values: five for categorizing the patient's injury severity, three for the LOS in the PICU, step-down unit, and floor units, and one for the patient's total LOS. RESULTS: A fuzzy ARTMAP neural network accurately categorizes 88% of mortality patients and 58.3% of critical PICU patients. A backpropagation neural network succeeded in predicting the total LOS to within 1 day for 51.4% and the ICU LOS to within 1 day for 70.4% of all evaluated patients. CONCLUSION: Information available in the first 10 mins of a patient's presentation at the emergency room can be used by an artificial neural network to predict injury severity and LOS. Artificial neural networks enable more effective resource planning and patient management.


Chronic nicotine administration exacerbates tau pathology in a transgenic model of Alzheimer's disease.
The association between nicotinic acetylcholine receptor (nAChR) dysfunction and cognitive decline in Alzheimer's disease (AD) has been widely exploited for its therapeutic potential. The effects of chronic nicotine exposure on Abeta accumulation have been studied in both humans and animal models, but its therapeutic efficacy for AD neuropathology is still unresolved. To date, no in vivo studies have addressed the consequences of activating nAChRs on tau pathology. To determine the effects of chronic nicotine administration on Abeta and tau pathology, we chronically administrated nicotine to a transgenic model of AD (3xTg-AD) in their drinking water. Here, we show that chronic nicotine intake causes an up-regulation of nicotinic receptors, which correlated with a marked increase in the aggregation and phosphorylation state of tau. These data show that nicotine exacerbates tau pathology in vivo. The increase in tau phosphorylation appears to be due to the activation of p38-mitogen-activated protein kinase, which is known to phosphorylate tau in vivo and in vitro. We also show that the 3xTg-AD mice have an age-dependent reduction of alpha7nAChRs compared with age-matched nontransgenic mice in specific brain regions. The reduction of alpha7nAChRs is first apparent at 6 months of age and is restricted to brain regions that show intraneuronal Abeta(42) accumulation. Finally, this study highlights the importance of testing compounds designed to ameliorate AD pathology in a model with both neuropathological lesions because of the differential effects it can have on either Abeta or tau.