Genomic structure and expression of the mouse growth factor receptor related to tyrosine kinases (Ryk).
We report the genomic organization of the mouse orphan receptor related to tyrosine kinases (Ryk), a structurally unclassified member of the growth factor receptor family. The mouse RYK protein is encoded by 15 exons distributed over a minimum of 81 kilobases. Genomic DNA sequences encoding a variant protein tyrosine kinase ATP-binding motif characteristic of RYK are unexpectedly found in two separate exons. A feature of the gene is an unmethylated CpG island spanning exon 1 and flanking sequences, including a TATA box-containing putative promoter and single transcription start site. Immunohistochemical examination of RYK protein distribution revealed widespread but developmentally regulated expression, which was spatially restricted within particular adult organs. Quantitative reduction of Southern blotting stringency for the detection of Ryk-related sequences provided evidence for a retroprocessed mouse pseudogene and a more distantly related gene paralogue. Extensive cross-species reactivity of a mouse Ryk kinase subdomain probe and the cloning of a Ryk orthologue from Caenorhabditis elegans demonstrate that Ryk and its relatives encode widely conserved members of a novel receptor tyrosine kinase subfamily.


Exploring the relationship between environmental enteric dysfunction and oral vaccine responses.
Oral vaccines significantly underperform in low-income countries. One possible contributory factor is environmental enteric dysfunction (EED), a subclinical disorder of small intestinal structure and function among children living in poverty. Here, we review studies describing oral vaccine responses and EED. We identified eight studies evaluating EED and oral vaccine responses. There was substantial heterogeneity in study design and few consistent trends emerged. Four studies reported a negative association between EED and oral vaccine responses; two showed no significant association; and two described a positive correlation. Current evidence is therefore insufficient to determine whether EED contributes to oral vaccine underperformance. We identify roadblocks in the field and future research needs, including carefully designed studies those can investigate this hypothesis further.


Therapeutic and adjunctive applications of an imidazoline anti-inflammatory agent.
A unique combination of anti-inflammatory and antiulcerogenic activities is described for 2-(2 methyl-4-chlorophenylamino)-2-imidazoline (CDMI). CDMI administered i.p. produced a dose-related decrease in aspirin-induced ulcers which persisted even in the presence of exogenously added HCI. The carrageenin-edema reducing activities of i.p. CDMI and p.o. aspirin were additive. When oral CDMI was combined with oral aspirin or oral indomethacin, the combinations also resulted in additive anti-inflammatory activities (80 and 94% vs. 52% for CDMI, 62% for aspirin and 71% for indomethacin alone). Moreover, gastric ulcerogenicity was reduced by 92% when either aspirin or indomethacin was combined with CDMI. These results indicate that CDMI and related compounds deserve further study as adjuncts to increase the margin of safety of currently used nonsteroidal anti-inflammatory agents. CDMI was also tested against a developing acute inflammatory reaction. When administered at 2 hr post carrageenin, CDMI was as effective as when it was administered 30 min before the carrageenin. These results are discussed as a possible reflection of an action on the lipoxygenase pathways of the arachidonic acid cascade that is not shared by the classical nonsteroidal anti-inflammatory agents.


Contributory and exacerbating roles of gaseous ammonia and organic dust in the etiology of atrophic rhinitis.
Pigs reared commercially indoors are exposed to air heavily contaminated with particulate and gaseous pollutants. Epidemiological surveys have shown an association between the levels of these pollutants and the severity of lesions associated with the upper respiratory tract disease of swine atrophic rhinitis. This study investigated the role of aerial pollutants in the etiology of atrophic rhinitis induced by Pasteurella multocida. Forty, 1-week-old Large White piglets were weaned and divided into eight groups designated A to H. The groups were housed in Rochester exposure chambers and continuously exposed to the following pollutants: ovalbumin (groups A and B), ammonia (groups C and D), ovalbumin plus ammonia (groups E and F), and unpolluted air (groups G and H). The concentrations of pollutants used were 20 mg m-3 total mass and 5 mg m-3 respirable mass for ovalbumin dust and 50 ppm for ammonia. One week after exposure commenced, the pigs in groups A, C, E, and G were infected with P. multocida type D by intranasal inoculation. After 4 weeks of exposure to pollutants, the pigs were killed and the extent of turbinate atrophy was assessed with a morphometric index (MI). Control pigs kept in clean air and not inoculated with P. multocida (group H) had normal turbinate morphology with a mean MI of 41.12% (standard deviation [SD], +/- 1. 59%). In contrast, exposure to pollutants in the absence of P. multocida (groups B, D, and F) induced mild turbinate atrophy with mean MIs of 49.65% (SD, +/-1.96%), 51.04% (SD, +/-2.06%), and 49.88% (SD, +/-3.51%), respectively. A similar level of atrophy was also evoked by inoculation with P. multocida in the absence of pollutants (group G), giving a mean MI of 50.77% (SD, +/-2.07%). However, when P. multocida inoculation was combined with pollutant exposure (groups A, C, and E) moderate to severe turbinate atrophy occurred with mean MIs of 64.93% (SD, +/-4.64%), 59.18% (SD, +/-2.79%), and 73.30% (SD, +/-3.19%), respectively. The severity of atrophy was greatest in pigs exposed simultaneously to dust and ammonia. At the end of the exposure period, higher numbers of P. multocida bacteria were isolated from the tonsils than from the nasal membrane, per gram of tissue. The severity of turbinate atrophy in inoculated pigs was proportional to the number of P. multocida bacteria isolated from tonsils (r2 = 0.909, P < 0.05) and nasal membrane (r2 = 0.628, P < 0.05). These findings indicate that aerial pollutants contribute to the severity of lesions associated with atrophic rhinitis by facilitating colonization of the pig's upper respiratory tract by P. multocida and also by directly evoking mild atrophy.


Crown ethers as a tool for the preparation of molecularly imprinted polymers.
Molecularly imprinted polymers have been prepared against aniline and a bis-aniline compound, making use of a crown ether (18-crown-6) to solubilize the monomer-template complexes. Subsequent chromatographic rebinding studies in the absence of crown ether revealed regioselectivity for the templates in the respective polymers. This study indicates that crown ethers can be potentially useful in conjunction with molecular imprinting to solubilize and imprint organic solvent-insoluble compounds.


Antiviral Treatments for Influenza.
Influenza is an acute respiratory illness caused by the influenza A, B, and C viruses. It can occur in local outbreaks or seasonal epidemics, with possibility to spread worldwide in a pandemic when a novel strain with significant antigenic differences emerges. During the past years, several new drugs have become available, with different accessibility related to specific countries' approval. We have conducted a review of literature, analyzing the most recent data on efficacy and safety of drugs currently available to treat influenza, with a particular attention toward special populations. Efficacy and safety profile of neuraminidase inhibitors (oseltamivir, zanamivir, laninamivir, peramivir) and recently approved cap-dependent endonuclease inhibitor baloxavir marboxil are reported in literature, but still little information is available about special populations such as critically ill patients and patients with a history of chronic respiratory disease. Moreover, the emergence of strains with reduced or no susceptibility to current drugs is a matter of concern, suggesting the need of constant monitoring of viral variants.


Isolated thoracic duct injury in penetrating neck trauma: a case report.
A 39-year-old man was admitted with a stab wound to left lower neck. Chest X-ray revealed a left hydropneumothorax. Thoracentesis was performed and analysis of the fluid revealed chyle. Patient was treated conservatively by closed chest drainage and total parenteral nutrition. On the basis of this clinical report and review of the literature, it is concluded that thoracic duct injury should be kept in mind in penetrating neck trauma and conservative treatment should be the first line therapy.


Role of contextual cues on the perception of spectrally reduced interrupted speech.
Understanding speech within an auditory scene is constantly challenged by interfering noise in suboptimal listening environments when noise hinders the continuity of the speech stream. In such instances, a typical auditory-cognitive system perceptually integrates available speech information and "fills in" missing information in the light of semantic context. However, individuals with cochlear implants (CIs) find it difficult and effortful to understand interrupted speech compared to their normal hearing counterparts. This inefficiency in perceptual integration of speech could be attributed to further degradations in the spectral-temporal domain imposed by CIs making it difficult to utilize the contextual evidence effectively. To address these issues, 20 normal hearing adults listened to speech that was spectrally reduced and spectrally reduced interrupted in a manner similar to CI processing. The Revised Speech Perception in Noise test, which includes contextually rich and contextually poor sentences, was used to evaluate the influence of semantic context on speech perception. Results indicated that listeners benefited more from semantic context when they listened to spectrally reduced speech alone. For the spectrally reduced interrupted speech, contextual information was not as helpful under significant spectral reductions, but became beneficial as the spectral resolution improved. These results suggest top-down processing facilitates speech perception up to a point, and it fails to facilitate speech understanding when the speech signals are significantly degraded.


Metabolomics in the study of spontaneous animal diseases.
Using analytical chemistry techniques such as nuclear magnetic resonance (NMR) spectroscopy and liquid or gas chromatography-mass spectrometry (LC/GC-MS), metabolomics allows detection of most endogenous and exogenous metabolites in a biological sample. Metabolomics has a wide range of applications, and has been employed in nutrition science, toxicology, environmental studies, and systems biology. Metabolomics is particularly useful in biomedical science, and has been used for diagnostic laboratory testing, identifying targets for drug development, and monitoring drug metabolism, mode of action, and toxicity. Despite its immense potential, metabolomics remains underutilized in the study of spontaneous animal diseases. Our aim was to comprehensively review the existing literature on the use of metabolomics in spontaneous veterinary diseases. Three databases were used to find journal articles that applied metabolomics in veterinary medicine. A screening process was then conducted to eliminate references that did not meet the eligibility criteria; only primary research studies investigating spontaneous animal disease were included; 38 studies met the inclusion criteria. The main techniques used were NMR and MS. All studies detected metabolite alterations in diseased animals compared with non-diseased animals. Metabolomics was mainly used to study diseases of the digestive, reproductive, and musculoskeletal systems. Inflammatory conditions made up the largest proportion of studies when articles were categorized by disease process. Following a comprehensive analysis of the literature on metabolomics in spontaneous veterinary diseases, we concluded that metabolomics, although in its early stages in veterinary research, is a promising tool regarding diagnosis, biomarker discovery, and in uncovering new insights into disease pathophysiology.


Quo vadis, Pep? Plant elicitor peptides at the crossroads of immunity, stress, and development.
The first line of inducible plant defence, pattern-triggered immunity (PTI), is activated by the recognition of exogenous as well as endogenous elicitors. Exogenous elicitors, also called microbe-associated molecular patterns, signal the presence of microbes. In contrast, endogenous elicitors seem to be generated and recognized under more diverse circumstances, making the evaluation of their biological relevance much more complex. Plant elicitor peptides (Peps) are one class of such endogenous elicitors, which contribute to immunity against attack by bacteria, fungi, as well as herbivores. Recent studies indicate that the Pep-triggered signalling pathways also operate during the response to a more diverse set of stresses including starvation stress. In addition, in silico data point to an involvement in the regulation of plant development, and a study on Pep-mediated inhibition of root growth supports this indication. Importantly, Peps are neither limited to the model plant Arabidopsis nor to a specific plant family like the previously intensively studied systemin peptides. On the contrary, they are present and active in angiosperms all across the phylogenetic tree, including many important crop plants. Here we summarize the progress made in research on Peps from their discovery in 2006 until now. We discuss the two main models which describe their likely function in plant immunity, highlight the studies supporting additional roles of Pep-triggered signalling and identify urgent research tasks to further uncover their biological relevance.


Sequence characteristics and phylogenetic analysis of H9N2 subtype avian influenza A viruses detected from poultry and the environment in China, 2018.
H9N2 subtype avian influenza A virus (AIV) is a causative agent that poses serious threats to both the poultry industry and global public health. In this study, we performed active surveillance to identify H9N2 AIVs from poultry (chicken, duck, and goose) and the environment of different regions in China, and we phylogenetically characterized the sequences. AIV subtype-specific reverse transcription polymerase chain reaction (RT-PCR) showed that 5.43% (83/1529) samples were AIV positive, and 87.02% (67/77) of which were H9N2 AIVs. Phylogenetic analysis revealed that all H9N2 field viruses belonged to the Y280-like lineage, exhibiting 93.9-100% and 94.6-100% of homology in the hemagglutinin (HA) gene and 94.4-100% and 96.3-100% in the neuraminidase (NA) gene, at the nucleotide (nt) and amino acid (aa) levels, respectively. All field viruses shared relatively lower identities with vaccine strains, ranging from 89.4% to 97.7%. The aa sequence at the cleavage site (aa 333-340) in HA of all the isolated H9N2 AIVs was PSRSSRG/L, which is a characteristic of low pathogenic avian influenza virus (LPAIV). Notably, all the H9N2 field viruses harbored eight glycosylation sites, whereas a glycosylation site 218 NRT was missing and a new site 313 NCS was inserted. All field viruses had NGLMR as their receptor binding sites (RBS) at aa position 224-229, showing high conservation with many recently-isolated H9N2 strains. All H9N2 field isolates at position 226 had the aa Leucine (L), indicating their ability to bind to sialic acid (SA) α, a 2-6 receptor of mammals that poses the potential risk of transmission to humans. Our results suggest that H9N2 AIVs circulating in poultry populations that have genetic variation and the potential of infecting mammalian species are of great significance when monitoring H9N2 AIVs in China.


Independent central and peripheral mediation of morphine-induced inhibition of gastrointestinal transit in rats.
The individual contributions of central (brain) and peripheral (enteric) sites in the mediation of the systemic actions of opioids are not well established. In this study, we made use of naltrexone methobromide, a quaternary analog of naltrexone, to separate the central and peripheral components of the slowing action of morphine on gastrointestinal transit in rats. It was established that i.c.v., but not s.c., administration of quaternary naltrexone antagonized morphine-induced analgesia in the radiant-heat tail-flick assay in rats. Thus, quaternary naltrexone probably does not enter the central nervous system in significant amounts after systemic administration. Systemic quaternary naltrexone antagonized, in a dose-related manner, the delaying effects of morphine on the movement of a charcoal meal along the gastrointestinal tract. Quaternary naltrexone was 30 or 100 times less potent than naltrexone when administered s.c. or i.c.v., respectively. Unlike naltrexone, quaternary naltrexone antagonized morphine-induced slowing of gastrointestinal transit only when administered by the same route (i.e., both s.c. or both i.c.v.). The apparent pA2 for s.c. quaternary naltrexone against s.c. morphine was not significantly different from the apparent pA2 for i.c.v. quaternary naltrexone against i.c.v. morphine. Distinct and independent central and peripheral systems appear to mediate morphine-induced inhibition of gastrointestinal transit in rats. However, the receptors are probably of the same type. Peripherally selective antagonists such as quaternary naltrexone may be useful in reversing morphine-induced inhibition of gastrointestinal transit without affecting analgesia.


Development and Progression of Diabetic Retinopathy in Adolescents and Young Adults With Type 2 Diabetes: Results From the TODAY Study.
The Treatment Options for Type 2 Diabetes in Adolescents and Youth (TODAY) study reported a 13.9% prevalence of diabetic retinopathy (DR) in youth with mean ± SD type 2 diabetes duration of 4.9 ± 1.5 years. After 7 years of additional follow-up, we report the risk factors for progression of DR in the TODAY cohort. Retinal photographs (n = 517) were obtained in 2010-2011 and again in 2017-2018 (n = 420) with standard stereoscopic seven-field digital fundus photography. Photographs were graded centrally using the Early Treatment Diabetic Retinopathy Study (ETDRS) scale. A total of 367 patients with gradable fundus photographs in at least one eye at both assessments were included in analyses of progression of DR, defined as an increase of three or more steps on the ETDRS scale. With mean ± SD age of 25.4 ± 2.5 years and diabetes duration of 12.0 ± 1.5 years, there was a 49% prevalence of any DR among participants. Prevalence by DR stage was as follows: 39% for very mild or mild nonproliferative DR (NPDR), 6% moderate to severe NPDR, and 3.8% proliferative DR. Compared with nonprogressors, participants who progressed three or more steps had significantly lower BMI, higher HbA1c, higher blood pressure, increased triglycerides, decreased C-peptide, and higher prevalence of other comorbidities. Multivariate analysis demonstrated that HbA1c was the dominant factor impacting DR progression. Poor glycemic control of youth-onset type 2 diabetes imparts a high risk for progression of DR, including advanced, sight-threatening disease by young adulthood.


Indocyanine green guided sentinel lymph node biopsy may have a high sensitivity for early (T1/T2) colon cancer: A prospective study in Indian patients.
Indocyanine green (ICG) dye guided near infrared fluorescence (NIR) imaging is a promising tool for mapping lymphatics. The aim of this study was to evaluate the role of ICG guided SLN biopsy in Indian colon cancer patients. Forty-eight patients of clinically staged T1-T3 node negative colon cancer underwent laparoscopic/open resection. Patients received colonoscopic peritumoral submucosal ICG injections for laparoscopic (n= 32) and subserosal injections for open resections (n= 16) followed by the detection of SLN using NIR camera. SLNs underwent conventional hematoxylin and eosin (H & E) staging with additional serial sectioning and immunohistochemistry for pancytokeratin antibody (ultra-staging). Detection rate and upstaging rate were the primary end points. Forty-eight patients were recruited. An average of 2.08 ± 1.27 SLNs were identified in 45 patients at a mean time of 8.2 ± 3.68 minutes with a detection rate of 93.75%. Mean age and mean BMI were 59.7 ± 12.54 years and 24.8 ± 4.09 kg/m2 , respectively. Eighteen patients had node positive disease, and SLN was false negative in four of these patients resulting in a sensitivity of 77.77% with a trend towards higher sensitivity for T1-T2 tumours (90% vs. 62.5%, p= 0.068). Upstaging rate was 10%. Negative predictive value (NPV) and accuracy of the procedure were 87.09% and 91.11%, respectively. ICG guided SLN biopsy can identify metastatic lymph nodes in colon cancer patients that can be missed on H & E staging with relatively higher sensitivity for early (T1/T2) tumours.


Gabapentin for Perioperative Pain Management for Uterine Aspiration: A Randomized Controlled Trial.
To evaluate the effect of oral gabapentin in conjunction with usual oral pain management regimens of lorazepam, ibuprofen, oxycodone, and acetaminophen for surgical abortion on pain 5 minutes postprocedure. This was a randomized, double-blind, placebo-controlled trial of patients from 6 0/7-14 6/7 weeks of gestation scheduled to undergo surgical abortion at the Duke Family Planning Clinic. Participants were administered 600 mg of oral gabapentin compared with placebo with usual oral pain management. Pain score was assessed using a 100-mm visual analog scale, with the primary outcome being pain score 5 minutes after the procedure. The effect of gabapentin was assessed using a linear regression model controlling for baseline pain. We also measured pain perception 24 hours after the procedure. Secondary outcome measures included anxiety, side effects, and usage of opiate pain medication in the 24-hour postoperative period. Out of 113 women screened for this study; 96 women were recruited, enrolled, and randomized to study treatment arm from August 2016 to June 2018. Pain at 5 minutes after the procedure was similar between the gabapentin and placebo groups ((Equation is included in full-text article.)=3.40; 95% CI -8.20 to 15.0; P=.56). Gabapentin and placebo were well tolerated, with no statistically significant difference in side effects or anxiety levels. Although prescription of opioids after the procedure was not standardized among patients, 73% of women received a short-term prescription for oxycodone. A lower percentage of women in the gabapentin group self-reported taking opioids in the 24 hours postprocedure (18% vs 47%; odds ratio 0.26; 95% CI 0.09-0.75). The addition of gabapentin to usual oral pain management regimens with paracervical block did not reduce postoperative pain for patients undergoing outpatient surgical abortion. Although the addition of gabapentin was well tolerated and reduced oral opiate use 24 hours postprocedure, it did not affect the experience of pain during and immediately after the procedure. ClinicalTrials.gov, NCT02725710.


Mouse H-2k-restricted cytotoxic T cells recognize antigenic determinants in both the HA1 and HA2 subunits of the influenza A/PR/8/34 hemagglutinin.
We have constructed two chimeric influenza hemagglutinin (HA) genes in which the HA1 and HA2 subunits of the HA molecule have been interchanged between influenza A/PR/8/34 (H1 subtype) and A/NT/60/68 (H3 subtype). These genes were used to construct recombinant vaccinia viruses that expressed intact chimeric HA. These recombinant viruses were used to test whether murine CTL recognize antigenic determinants in either the HA1, HA2, or both subunits. We found that both subunits of the HA molecule contain determinants for CTL. This implies that CTL have, at least in part, separate antigenic determinants from B lymphocytes, which recognize mainly epitopes within the HA1 subunit.


Indirect effects of predators control herbivore richness and abundance in a benthic eelgrass (Zostera marina) mesograzer community.
Herbivore communities can be sensitive to changes in predator pressure (top-down effects) and resource availability (bottom-up effects) in a wide range of systems. However, it remains unclear whether such top-down and bottom-up effects reflect direct impacts of predators and/or resources on herbivores, or are indirect, reflecting altered interactions among herbivore species. We quantified direct and indirect effects of bottom-up and top-down processes on an eelgrass (Zostera marina) herbivore assemblage. In a field experiment, we factorially manipulated water column nutrients (with Osmocote(™) slow-release fertilizer) and predation pressure (with predator exclusion cages) and measured the effects on herbivore abundance, richness and beta diversity. We examined likely mechanisms of community responses by statistically exploring the response of individual herbivore species to trophic manipulations. Predators increased herbivore richness and total abundance, in both cases through indirect shifts in community composition. Increases in richness occurred through predator suppression of common gammarid amphipod species (Monocorophium acherusicum and Photis brevipes), permitting the inclusion of rarer gammarid species (Aoroides columbiae and Pontogeneia rostrata). Increased total herbivore abundance reflected increased abundance of a caprellid amphipod species (Caprella sp.), concurrent with declines in the abundance of other common species. Furthermore, predators decreased beta diversity by decreasing variability in Caprella sp. abundance among habitat patches. Osmocote(™) fertilization increased nutrient concentrations locally, but nutrients dissipated to background levels within 3 m of the fertilizer. Nutrient addition weakly affected the herbivore assemblage, not affecting richness and increasing total abundance by increasing one herbivore species (Caprella sp.). Nutrient addition did not affect beta diversity. We demonstrated that assemblage-level effects of trophic manipulations on community structure are the result of distinct and often indirect responses of herbivore species. These results underscore the importance of understanding herbivore-herbivore interactions in a system commonly subjected to both eutrophication and overfishing.


Identification and analysis of the polyhydroxyalkanoate-specific beta-ketothiolase and acetoacetyl coenzyme A reductase genes in the cyanobacterium Synechocystis sp. strain PCC6803.
Synechocystis sp. strain PCC6803 possesses a polyhydroxyalkanoate (PHA)-specific beta-ketothiolase encoded by phaA(Syn) and an acetoacetyl-coenzyme A (CoA) reductase encoded by phaB(Syn). A similarity search of the entire Synechocystis genome sequence identified a cluster of two putative open reading frames (ORFs) for these genes, slr1993 and slr1994. Sequence analysis showed that the ORFs encode proteins having 409 and 240 amino acids, respectively. The two ORFs are colinear and most probably coexpressed, as revealed by sequence analysis of the promoter regions. Heterologous transformation of Escherichia coli with the two genes and the PHA synthase of Synechocystis resulted in accumulation of PHAs that accounted for up to 12.3% of the cell dry weight under high-glucose growth conditions. Targeted disruption of the above gene cluster in Synechocystis eliminated the accumulation of PHAs. ORFs slr1993 and slr1994 thus encode the PHA-specific beta-ketothiolase and acetoacetyl-CoA reductase of Synechocystis and, together with the recently characterized PHA synthase genes in this organism (S. Hein, H. Tran, and A. Steinbüchel, Arch. Microbiol. 170:162-170, 1998), form the first complete PHA biosynthesis pathway known in cyanobacteria. Sequence alignment of all known short-chain-length PHA-specific acetoacetyl-CoA reductases also suggests an extended signature sequence, VTGXXXGIG, for this group of proteins. Phylogenetic analysis further places the origin of phaA(Syn) and phaB(Syn) in the gamma subdivision of the division Proteobacteria.


Policy changes to implement intramural sports in North Carolina middle schools: simulated effects on sports participation rates and physical activity intensity, 2008-2009.
Extracurricular school sports programs can provide adolescents, including those who are economically disadvantaged, with opportunities to engage in physical activity. Although current models favor more exclusionary interscholastic sports, a better understanding is needed of the potential effects of providing alternative school sports options, such as more inclusive intramural sports. The purpose of this study was to simulate the potential effect of implementing intramural sports programs in North Carolina middle schools on both the rates of sports participation and on energy expenditure related to physical activity levels. Simulations were conducted by using a school-level data set developed by integrating data from multiple sources. Baseline rates of sports participation were extrapolated from individual-level data that were based on school-level characteristics. A regression model was estimated by using the simulated baseline school-level sample. Participation rates and related energy expenditure for schools were calculated on the basis of 2 policy change scenarios. Currently, 37.2% of school sports participants are economically disadvantaged. Simulations suggested that policy changes to implement intramural sports along with interscholastic sports could result in more than 43,000 new sports participants statewide, of which 64.5% would be economically disadvantaged students. This estimate represents a 36.75% increase in economically disadvantaged participants. Adding intramural sports to existing interscholastic sports programs at all middle schools in North Carolina could have an annual effect of an additional 819,892.65 kilogram calories expended statewide. Implementing intramural sports may provide economically disadvantaged students more access to sports, thus reducing disparities in access to school sports while increasing overall physical activity levels among all children.


Wear studies of all UHMWPE couples under various bio-tribological conditions.
Wear tests were undertaken in which ultra high molecular weight polyethylene (UHMWPE) was rubbed against itself. Tests primarily employed a pin-on-plate wear test machine, with distilled water, Ringer solution and dilute bovine serum being used as the lubricants. Loads of 10N and 40N were employed, and some test pins had a rotational motion added. In all cases wear was high, with mean wear factors of up to 91 10 -6 mm3/Nm being measured, but the addition of rotation reduced the amount of material worn from the test plates. In the presence of bovine serum and under reciprocation only, pin wear was relatively low. With bovine serum as the lubricant, total mean wear factors for the UHMWPE couples were calculated to be in the range of 35 to 58 10-6mm3/Nm. Therefore the pin-on-plate tests showed that the choice of lubricant as well as the motion applied to the test pin had a significant influence on the wear volumes measured. A two-piece UHMWPE 'prosthesis' with matching hemispherical faces was fabricated and tested on a finger simulator. Distilled water was used as the lubricant and wear factors were found to be greater for the metacarpal component, 21 10 -6mm3/Nm, than the phalangeal component, 3 10-6mm3/Nm, after ten million cycles of testing. This result paralleled the greater wear seen by the plate than by the pin in the pin-on-plate tests under reciprocating motion. (Journal of Applied Biomaterials & Biomechanics 2004; 2: 29-34).


Reliance on external cues during serial sequential movement in major depression.
Maintenance of motor set in patients with unipolar major depression was examined. Twelve melancholic and 12 non-melancholic depressed patients and 24 age matched controls performed a serial choice reaction time task while external cues aiding maintenance of a motor set were systematically removed. Melancholic patients were significantly slower than controls with no reduction in external cues and with a moderate reduction in external cues. At a high level of reduction in external cues, seven of 12 melancholic patients (but only three of 12 non-melancholic patients and controls) were unable to complete the task; suggesting a greater reliance on external cues, perhaps implicating a failure of motor planning ability in melancholic patients. This, in turn, may point to a prefrontal (premotor) deficit in melancholic depression, with possible commonalities with Parkinson's disease.


Effect of dietary alpha-linolenate on platelet-activating factor production in rat peritoneal polymorphonuclear leukocytes.
The effects of dietary alpha-linolenate (18:3, n-3) and linoleate (18:2, n-6) on platelet-activating factor (PAF) production were examined. Rats were fed an alpha-linolenic acid-rich (perilla oil) diet or a linoleic acid-rich (safflower oil) diet for 6 wk, and polymorphonuclear leukocytes (PMN) were elicited by peritoneal injection of casein. The overall phospholipid content and composition as well as the subclass distribution of choline and ethanolamine glycerophospholipids in PMN were not altered by these diets. However, with the perilla oil diet their content of a putative precursor of PAF, 1-alkyl-2-arachidonoyl-sn-glycero-3-phosphocholine was approximately 50% of that with safflower oil diet. On exposure to various concentrations of FMLP, PAF formation by PMN in the perilla oil group was less than 50% of that by PMN in the safflower oil group. A larger difference in PAF productions by PMN in the two dietary groups was observed on their stimulation with calcium ionophore A23187. These results demonstrate that PAF production is modulated in some as yet unknown way by changing the alpha-linolenate/linoleate balance of the diet.


Characterization of a newly developed chicken 44K Agilent microarray.
The development of microarray technology has greatly enhanced our ability to evaluate gene expression. In theory, the expression of all genes in a given organism can be monitored simultaneously. Sequencing of the chicken genome has provided the crucial information for the design of a comprehensive chicken transcriptome microarray. A long oligonucleotide microarray has been manually curated and designed by our group and manufactured using Agilent inkjet technology. This provides a flexible and powerful platform with high sensitivity and specificity for gene expression studies. A chicken 60-mer oligonucleotide microarray consisting of 42,034 features including the entire Marek's disease virus, two avian influenza virus (H5N2 and H5N3), and 150 chicken microRNAs has been designed and tested. In an important validation study, total RNA isolated from four major chicken tissues: cecal tonsil (C), ileum (I), liver (L), and spleen (S) were used for comparative hybridizations. More than 95% of spots had high signal noise ratio (SNR > 10). There were 2886, 2660, 358, 3208, 3355, and 3710 genes differentially expressed between liver and spleen, spleen and cecal tonsil, cecal tonsil and ileum, liver and cecal tonsil, liver and ileum, spleen and ileum (P < 10-7), respectively. There were a number of tissue-selective genes for cecal tonsil, ileum, liver, and spleen identified (95, 71, 535, and 108, respectively; P < 10-7). Another highlight of these data revealed that the antimicrobial peptides GAL1, GAL2, GAL6 and GAL7 were highly expressed in the spleen compared to other tissues tested. A chicken 60-mer oligonucleotide 44K microarray was designed and validated in a comprehensive survey of gene expression in diverse tissues. The results of these tissue expression analyses have demonstrated that this microarray has high specificity and sensitivity, and will be a useful tool for chicken functional genomics. Novel data on the expression of putative tissue specific genes and antimicrobial peptides is highlighted as part of this comprehensive microarray validation study. The information for accessing and ordering this 44K chicken array can be found at http://people.tamu.edu/~hjzhou/TAMUAgilent44KArray/


Incorporating methods and findings from neuroscience to better understand placebo and nocebo effects in sport.
Placebo and nocebo effects are a factor in sports performance. However, the majority of published studies in sport science are descriptive and speculative regarding mechanisms. It is therefore not unreasonable for the sceptic to argue that placebo and nocebo effects in sport are illusory, and might be better explained by variations in phenomena such as motivation. It is likely that, in sport at least, placebo and nocebo effects will remain in this empirical grey area until researchers provide stronger mechanistic evidence. Recent research in neuroscience has identified a number of consistent, discrete and interacting neurobiological and physiological pathways associated with placebo and nocebo effects, with many studies reporting data of potential interest to sport scientists, for example relating to pain, fatigue and motor control. Findings suggest that placebos and nocebos result in activity of the opioid, endocannabinoid and dopamine neurotransmitter systems, brain regions including the motor cortex and striatum, and measureable effects on the autonomic nervous system. Many studies have demonstrated that placebo and nocebo effects associated with a treatment, for example an inert treatment presented as an analgesic or stimulant, exhibit mechanisms similar or identical to the verum or true treatment. Such findings suggest the possibility of a wide range of distinct placebo and nocebo mechanisms that might influence sports performance. In the present paper, we present some of the findings from neuroscience. Focussing on fatigue as an outcome and caffeine as vehicle, we propose three approaches that researchers in sport might incorporate in their studies in order to better elucidate mechanisms of placebo/nocebo effects on performance.


Events following the infections of enucleate cells with measles virus.
The development of measles virus (Edmonston) and SSPE measles virus (Horta-Barbosa) has been examined in enucleate BSC 1 cells. New antigen synthesis in measles virus infected enucleate cells has been demonstrated by fluorescent antibody, by the formation of extensive syncytia from enucleate cells alone and by analysis of polypeptide formation by polyacrylamide gel electrophoresis. All polypeptides formed in nucleate cells were also present in enucleate cells but the amount synthesized was reduced to around 20% of that in nucleate cells. There was also a significant reduction in the amount of antigen detected by fluorescent antibody in enucleate as compared to nucleate preparations. Examination of RNA synthesis in infected enucleate cells revealed only a marginal increase in acid-insoluble material. Titration of the output of infectious virus from enucleate cells infected at both 37 and 31 degrees C indicated a consistent reduction of almost two log units compared to nucleate cells. That the enucleate cells were capable of replicating input genome at these times was demonstrated by the successful growth of respiratory syncytial virus, both at 37 and 31 degrees C. SSPE measles virus grew to higher yield in nucleate BSC 1 than measles virus but there was again a reduction of more than two log units in enucleate cells. All polypeptides synthesized in SSPE infected nucleate cells were apparent in enucleate cells.


Leucine stimulates translation initiation in skeletal muscle of postabsorptive rats via a rapamycin-sensitive pathway.
The objectives of the present study were twofold: 1) to determine whether leucine is unique among the branched-chain amino acids (BCAA) in its ability to stimulate protein synthesis in skeletal muscle of food-deprived rats; and 2) to investigate whether changes in muscle protein synthesis after leucine administration involve a signaling pathway that includes the protein kinase mammalian target of rapamycin (mTOR). In the first set of experiments, food-deprived (18 h) male rats (200 g) were orally administered saline or 270 mg valine, isoleucine or leucine. In the second set of experiments, food-deprived rats were injected intravenously with rapamycin (0.75 mg/kg), a specific inhibitor of mTOR, before leucine administration. Only leucine stimulated protein synthesis in skeletal muscle above saline-treated controls (P: < 0.05). Furthermore, leucine was most effective among the BCAA at enhancing phosphorylation of eukaryotic initiation factor (eIF), 4E binding protein 1 (4E-BP1) and the 70-kDa ribosomal protein S6 kinase (S6K1). Leucine-dependent hyperphosphorylation of 4E-BP1 increased the availability of eIF4E to form the active eIF4G.eIF4E complex. To a lesser extent, isoleucine also enhanced phosphorylation of 4E-BP1 and S6K1. Rapamycin inhibited protein synthesis in both leucine-treated and food-deprived rats. Additionally, rapamycin prevented the stimulatory effects of leucine on eIF4E availability for binding eIF4G and inhibited leucine-dependent phosphorylation of S6K1. The data demonstrate that leucine is unique among the BCAA in its ability to stimulate protein synthesis in muscle of food-deprived rats. We show for the first time that leucine-dependent stimulation of translation initiation in vivo occurs via a rapamycin-sensitive pathway.


Drainage of cerebral interstitial fluid into deep cervical lymph of the rabbit.
Lymph from the jugular lymph trunks of anesthetized rabbits has been continuously collected and radioiodinated albumin (RISA) therein estimated after microinjection of 1 microliter of 131I-albumin into the caudate nucleus, after single intraventricular injections, and during intraventricular infusions. Comparison of lymph at 7 and 25 h after intracerebral microinjection with efflux of radioactivity from whole brain suggests that about 50% of cleared radioactivity goes through lymph. Concentrations, normalized to cerebrospinal fluid (CSF), were much higher in lymph and retropharyngeal nodes after brain injection than after CSF injection or infusion. Also after brain injection, lymph and nodes contained more activity on injected side in contrast to lack of laterality after CSF administration. Calculation suggests that less than 30% of RISA cleared from brain can do so via a pool of well-mixed CSF. Analysis of tissues is compatible with much RISA draining by bulk flow via cerebral perivascular spaces plus passage from subarachnoid space of olfactory lobes into submucous spaces of nose and thus to lymph.


Spontaneous abscess of the lumbar spine presenting as subacute back pain.
A 52-year-old lady was admitted with back pain, fever and reduced sensation in her left leg. Soon after admission she developed acute urinary retention. She underwent urgent MRI of the spine which showed an epidural abscess compressing the thecal sac which was treated with neurosurgical decompression and intravenous/oral antibiotics. Cultures from theatre grew Staphylococcus aureus sensitive to flucloxacillin. After 12 weeks of intravenous/oral antibiotics, she was discharged without any neurological disability.


Uniform P doped Co-Ni-S nanostructures for asymmetric supercapacitors with ultra-high energy densities.
Uniform P doped Co-Ni-S nanosheet arrays were directly grown on Ni foams by an efficient and cost-effective process. The binder-free electrode of P doped Co-Ni-S nanosheet arrays possesses an ultra-high specific capacitance of ∼3677 F g-1 at 1 A g-1 with an excellent rate capability (∼63% capacitance retention at 20 A g-1) and considerable cycling performance (∼84% capacitance retention after 10 000 cycles). Correspondingly, the asymmetric supercapacitors assembled with P doped Co-Ni-S as the positive electrode and AC as the negative electrode display an ultra-high energy density of ∼68.7 W h kg-1 at a power density of ∼0.8 kW kg-1. In view of these features, this work provides a simple and scalable strategy for designing electrodes and devices with superior electrochemical performance in next generation energy storage applications.


Effects of programming intake on performance and carcass characteristics of feedlot cattle.
We conducted two experiments to determine the effects of various intake restriction strategies on performance and carcass characteristics of feedlot cattle. In Exp. 1, 104 steer calves (273 +/- 12 kg BW) were allotted to 12 pens. During Period 1 (273 to 372 kg BW), steers were limit-fed according to net energy equations to achieve predicted gains of .91, 1.13, or 1.36 kg/d. Control steers were offered ad libitum access to feed during this period. During Period 2 (372 to 535 kg BW), all steers were offered ad libitum access to feed. In Period 1, steers limit-fed for predicted gains of .91, 1.13, and 1.36 kg/d gained 1.03, 1.22, and 1.40 kg/d, respectively. Control steers offered ad libitum access to feed gained 1.66 kg/d. Steers that were limit-fed in Period 1 were able to compensate in Period 2; for the total experiment, there were no differences (P > .10) among the four feeding strategies investigated for rate of gain, feed efficiency, total feed intake, and carcass characteristics. Experiment 2 used 107 steer calves (300 +/- 11 kg BW). Four feeding strategies were compared: step-wise increases in intake to program for increasing rate of gain, stepwise decreases in intake to program for decreasing rate of gain, feeding to hold gain constant at 1.36 kg/d, or offering steers ad libitum access to feed throughout the experiment to allow for maximum gains. When averaged over the total experiment, growth rate and days on feed were not affected (P > .10) by feeding system. Steers fed to achieve a step-wise increase in growth rate throughout the experiment had the lowest (P < .09) daily intakes and the highest (P < .09) feed efficiencies. Steers fed for increasing gains required 109 kg/steer less (P < .09) total feed to reach market weight than those offered ad libitum access to feed throughout the experiment. Feeding strategy had little effect on carcass characteristics. We concluded that as intake is restricted to a greater extent, net energy equations are less accurate in predicting rate of gain. Programming intake during the feeding period can result in significant reductions in feed expenditures.


Use of Psychotropic Medications and Visits to Psychiatrists and Psychologists among Individuals with Nonsyndromic Oral Clefts: A Population-Based Cohort Study.
Oral clefts (OCs) are among the most common congenital malformations and can have a large impact on the life of the affected individual. Research findings regarding the psychological and psychosocial consequences of OC are inconclusive. Using Danish nationwide registers, we investigated redeemed prescriptions of psychotropic medication during 1996 to 2012 and visits to psychiatrists and psychologists during 1996 to 2011 among individuals born with nonsyndromic OC in Denmark between 1936 and 2009 and a comparison cohort of individuals without OC. This includes 8244 individuals with OC and 82,665 individuals without OC. The Cox regression analysis revealed 12% (95% confidence interval [CI], 7 to 16%) increased risk of using any psychotropic medication for individuals with OC. When examining by cleft type, higher risks for medication use were observed in individuals with cleft lip and palate (CLP) or cleft palate (CP) only. The largest increased relative risk was found for use of antipsychotics and stimulants for individuals with CP followed by use of antipsychotics for individuals with CLP. We found increased risk of visits to psychiatrists for individuals with CP and no increased risk for visits to psychologists for either group. This study indicates that a small group of individuals with nonsyndromic OC, in particular those with palatal involvement, have greater risk of using psychotropic medications. However, elevated use was also observed among younger individuals with cleft lip (CL) only. There seems to be only a modest increase in visits to health professionals for psychological reasons. Undiagnosed syndromes (e.g., 22q11 deletion syndrome), may, however, contribute to an overestimation of the associations. Birth Defects Research 109:824-835, 2017. © 2017 Wiley Periodicals, Inc.


How should we normalize electromyograms obtained from healthy participants? What we have learned from over 25 years of research.
Electromyograms (EMGs) need to be normalized if comparisons are sought between trials when electrodes are reapplied, as well as between different muscles and individuals. The methods used to normalize EMGs recorded from healthy individuals have been appraised for more than a quarter of a century. Eight methods were identified and reviewed based on criteria relating to their ability to facilitate the comparison of EMGs. Such criteria included the magnitude and pattern of the normalized EMG, reliability, and inter-individual variability. If the aim is to reduce inter-individual variability, then the peak or mean EMG from the task under investigation should be used as the normalization reference value. However, the ability of such normalization methods to facilitate comparisons of EMGs is questionable. EMGs from MVCs can be as reliable as those from submaximal contractions, and do not appear to be affected by contraction mode or joint kinematics, particularly for the elbow flexors. Thus, the EMG from an isometric MVC is endorsed as a normalization reference value. Alternatively the EMG from a dynamic MVC can be used, although it is recognized that neither method is guaranteed to be able to reveal how active a muscle is in relation to its maximal activation capacity.


Exploring the relationship between underlying dimensions of posttraumatic stress disorder and depression in a national, trauma-exposed military sample.
Posttraumatic stress disorder (PTSD) and depression are highly comorbid and intercorrelated. Yet little research has examined the underlying processes explaining their interrelationship. In the present survey study, the investigators assessed the combined symptom structure of PTSD and depression symptoms, to examine shared, underlying psychopathological processes. Participants included 740 Canadian military veterans from a national, epidemiological survey, previously deployed on peacekeeping missions and administered the PTSD Checklist and Center for Epidemiological Studies-Depression Scale (CES-D). An eight-factor PTSD/depression model fit adequately. In analyses validating the structure, PTSD's dysphoria factor was more related to depressive affect than to several other PTSD and depression factors. Somatic problems were more related to dysphoria than to other PTSD factors. Only military veterans were sampled, and without the use of structured diagnostic interviews. Results highlight a set of interrelationships that PTSD's dysphoria factor shares with specific depression factors, shedding light on the underlying psychopathology of PTSD that emphasizes dysphoric mood.


Clinical Utility of a Unique Genome-Wide DNA Methylation Signature for KMT2A-Related Syndrome.
Wiedemann-Steiner syndrome (WDSTS) is a Mendelian syndromic intellectual disability (ID) condition associated with hypertrichosis cubiti, short stature, and characteristic facies caused by pathogenic variants in the KMT2A gene. Clinical features can be inconclusive in mild and unusual WDSTS presentations with variable ID (mild to severe), facies (typical or not) and other associated malformations (bone, cerebral, renal, cardiac and ophthalmological anomalies). Interpretation and classification of rare KMT2A variants can be challenging. A genome-wide DNA methylation episignature for KMT2A-related syndrome could allow functional classification of variants and provide insights into the pathophysiology of WDSTS. Therefore, we assessed genome-wide DNA methylation profiles in a cohort of 60 patients with clinical diagnosis for WDSTS or Kabuki and identified a unique highly sensitive and specific DNA methylation episignature as a molecular biomarker of WDSTS. WDSTS episignature enabled classification of variants of uncertain significance in the KMT2A gene as well as confirmation of diagnosis in patients with clinical presentation of WDSTS without known genetic variants. The changes in the methylation profile resulting from KMT2A mutations involve global reduction in methylation in various genes, including homeobox gene promoters. These findings provide novel insights into the molecular etiology of WDSTS and explain the broad phenotypic spectrum of the disease.


Identification of putative candidate genes for juvenile wood density in Pinus radiata.
Wood formation is a complex developmental process driven by the annual activity of the vascular cambium. Conifers usually produce juvenile wood at young ages followed by mature wood for the rest of their lifetime. Juvenile wood exhibits poorer wood quality (i.e., lower density) compared with mature wood and can account for up to 50% of short-rotation harvested logs, thus representing a major challenge for commercial forestry globally. Wood density is an important quality trait for many timber-related products. Understanding the molecular mechanisms involved in the regulation of juvenile wood density is critical for the improvement of juvenile wood quality via marker-aided selection. A previous study has identified several candidate genes affecting mature wood density in Picea sitchensis (Bong.) Carr.; however, genes associated with juvenile wood density in conifers remain poorly characterized. Here, cDNA microarrays containing 3320 xylem unigenes were used to investigate genes differentially transcribed in juvenile wood with high (HD) and low density (LD) in Pinus radiata D.Don. In total, 814 xylem unigenes with differential transcription were identified in at least one of two microarray experiments and 73 genes (45 for HD, 28 for LD) were identified in both experiments, thus representing putative candidate genes for juvenile wood density. Interestingly, cellulose synthases (PrCesA3, PrCesA11) and sucrose synthase (SuSy), which are involved in secondary cell wall formation, had stronger transcription in juvenile wood with HD, while genes functioning in primary wall formation (pectin synthesis, cell expansion and other modifications) were more transcribed in LD wood. Cell wall genes encoding monolignol biosynthesis enzymes, arabinogalactan proteins, actins and tubulins were differentially transcribed in either HD or LD juvenile wood; however, the latter had exclusively greater transcription of genes involved in monolignol polymerization (laccase and peroxidase). The identified candidate genes also included many non-cell-wall genes (transcription factors, environmental-responsive genes, hormone signalling, etc.) and genes with unknown functions, suggesting complex gene pathways in the regulation of juvenile wood density. Interestingly, 19 out of 73 candidate genes for wood density were among the 108 candidate genes previously identified for microfibril angle, and 16 genes appeared to influence both traits in a synergistic manner for wood stiffness.


Boosting epidermal growth factor receptor expression by gene gun transfection stimulates epidermal growth in vivo.
Expression constructs encoding a full-length cDNA encoding the human epidermal growth factor receptor, or reporter gene for green fluorescent protein or luciferase were coated onto gold particles and driven into porcine skin using a gene gun delivery system. Strategies for epidermal growth factor receptor boosting were tested in two types of wounds. For grafted wounds, intact porcine skin was pretreated by the introduction of the epidermal growth factor receptor expression construct 24 hours before its harvesting as a split-thickness skin graft. Partial-thickness excisional wound beds (donor sites) were transfected at the time of their creation. Wound healing parameters were subsequently tested in the presence or absence of excess epidermal growth factor ligand. Initial distributions of gene gun delivered gold particles as well as luciferase expression levels suggested that optimal skin penetrations and expression levels were achieved at 500 psi for intact epidermis and 300 psi for exposed wound beds. At 2 days after gene delivery, visualization of green fluorescent protein by fluorescence microscopy showed focal expression of green fluorescent protein at the advancing epithelial outgrowths found at wound edges or surviving epithelial remnants. Green fluorescent protein expression appeared transient since no green fluorescent protein was noted in specimens removed at 4 days after injury. Northern blot analysis on mRNA isolated from wounds 2 days after introduction of epidermal growth factor receptor coated gold particles by gene gun confirmed the expression of the human epidermal growth factor receptor transgene in both skin grafts and excisional wounds. Skin grafts showed subsequent biological responses to the introduction of excessive epidermal growth factor receptor as well as expression of the human epidermal growth factor receptor construct within healing epidermis. While control autografts (reporter gene treated, epidermal growth factor alone, placebo formula, no treatment) showed few 5'-bromodeoxyuridine-labeled cells, epidermal growth factor receptor autografts showed 5'-bromodeoxyuridine labeling of nearly every basal cell. Favorable wound healing outcomes were also shown within excisional wounds following in vivo boosting of epidermal growth factor receptor. Four days after receiving epidermal growth factor receptor particle growth factor receptor transgene. Application of topical epidermal growth factor ligand resulted in the highest percentage of resurfacing. Maximal re-epithelialization was noted in wound beds receiving both receptor boosting and excessive daily epidermal growth factor ligand. A modest increase in the thickness of the granulation tissue followed gene therapy with epidermal growth factor receptor. In summary these in vivo data suggest that it is possible to boost in vivo expression of a tyrosine kinase receptor during wound repair. Increased epidermal growth factor receptor expression has an integral impact on cell proliferation, rates of resurfacing and dermal components and merits consideration as a possible therapeutic agent.


Identification of the Asymptomatic Patient With Severe Mitral Regurgitation: Discrepancy Between Research and Clinical Practice.
Organic mitral regurgitation (MR) is a common disorder, and because of the increase in population and its aging, the occurrence of MR is steadily increasing. Current guideline recommendations on the management of asymptomatic severe MR are conflicting and based solely on registries or nonrandomized trials in expert heart valve clinics, resulting in a lack of evidence for the best treatment strategy. In this review, we will evaluate the latest evidence on diagnostic approaches and treatment strategies for asymptomatic patients without a clear indication for surgical intervention. Implications for management in daily practice are discussed, including an update on the diagnostic approaches that are currently available for the evaluation of MR. For optimal care, it is important that every severe MR patient, including the unidentified patient, is referred to a specialized heart team and is assessed on an individual basis according to the guideline recommendations, experience of the surgical center, and the patient's characteristics and preferences. Screening and diagnostic approaches need to be performed on the basis of standardized protocols and strict criteria. In addition, specialized valve centers must meet the surgical criteria to guarantee high reparability rates in asymptomatic patients. Awareness among cardiologists and cardiothoracic surgeons, improved guidelines adherence, and a systematic approach, including strict criteria in the management of asymptomatic patients with severe organic MR, will ensure reliable and applicable results in research and daily clinical practice.


Kinetics of concomitant transfer and hydrolysis reactions catalysed by the exocellular DD-carboxypeptidase-transpeptidase of streptomyces R61.
When Ac(2)-l-Lys-d-Ala-d-Ala and either meso-diaminopimelic acid or Gly-l-Ala are exposed to the exocellular dd-carboxypeptidase-transpeptidase of Streptomyces R61, transpeptidation reactions yielding Ac(2)-l-Lys-d-Ala-(d)-meso- diaminopimelic acid and Ac(2)-l-Lys-d-Ala-Gly-l-Ala occur concomitantly with the hydrolysis of the tripeptide into Ac(2)-l-Lys-d-Ala. The proportion of the enzyme activity which can be channelled in the transpeptidation and the hydrolysis pathways depends upon the pH and the polarity of the environment. Transpeptidation is favoured both by increasing the pH and by decreasing the water content of the reaction mixtures. Kinetics suggest that the reactions proceed through an ordered mechanism in which the acceptor molecule (meso-diaminopimelic acid or Gly-l-Ala) binds first to the enzyme. Both acceptors behave as non-competitive inhibitors of the hydrolysis pathway. Transpeptidation is inhibited by high concentrations of Gly-l-Ala but not by high concentrations of meso-diaminopimelic acid. The occurrence on the enzyme of an additional inhibitory binding site for Gly-l-Ala is suggested.


Activation of pre-synaptic M-type K+ channels inhibits [3H]D-aspartate release by reducing Ca2+ entry through P/Q-type voltage-gated Ca2+ channels.
In this study, the functional consequences of the pharmacological modulation of the M-current (I(KM)) on cytoplasmic Ca(2+) intracellular Ca(2+)concentration ([Ca(2+)](i)) changes and excitatory neurotransmitter release triggered by various stimuli from isolated rat cortical synaptosomes have been investigated. K(v)7.2 immunoreactivity was identified in pre-synaptic elements in cortical slices and isolated glutamatergic cortical synaptosomes. In cerebrocortical synaptosomes exposed to 20 mM [K(+)](e), the I(KM) activator retigabine (RT, 10 microM) inhibited [(3)H]D-aspartate ([(3)H]D-Asp) release and caused membrane hyperpolarization; both these effects were prevented by the I(KM) blocker XE-991 (20 microM). The I(KM) activators RT (0.1-30 microM), flupirtine (10 microM) and BMS-204352 (10 microM) inhibited 20 mM [K(+)](e)-induced synaptosomal [Ca(2+)](i) increases; XE-991 (20 microM) abolished RT-induced inhibition of depolarization-triggered [Ca(2+)](i) transients. The P/Q-type voltage-sensitive Ca(2+)channel (VSCC) blocker omega-agatoxin IVA prevented RT-induced inhibition of depolarization-induced [Ca(2+)](i) increase and [(3)H]D-Asp release, whereas the N-type blocker omega-conotoxin GVIA failed to do so. Finally, 10 microM RT did not modify the increase of [Ca(2+)](i) and the resulting enhancement of [(3)H]D-Asp release induced by [Ca(2+)](i) mobilization from intracellular stores, or by store-operated Ca(2+)channel activation. Collectively, the present data reveal that the pharmacological activation of I(KM) regulates depolarization-induced [(3)H]D-Asp release from cerebrocortical synaptosomes by selectively controlling the changes of [Ca(2+)](i) occurring through P/Q-type VSCCs.


Ab initio theoretical study of non-covalent adsorption of aromatic molecules on boron nitride nanotubes.
We have studied non-covalent functionalization of boron nitride nanotubes (BNNTs) with benzene molecule and with seven other different heterocyclic aromatic rings (furan, thiophene, pyrrole, pyridine, pyrazine, pyrimidine, and pyridazine, respectively). A hybrid density functional theory (DFT) method with the inclusion of dispersion correction is employed. The structural and electronic properties of the functionalized BNNTs are obtained. The DFT calculation shows that upon adsorption to the BNNT, the center of aromatic rings tend to locate on top of the nitrogen site. The trend of adsorption energy for the aromatic rings on the BNNTs shows marked dependence on different intermolecular interactions, including the dispersion interaction (area of the delocalized π bond), the dipole-dipole interaction (polarization), and the electrostatic repulsion (lone pair electrons). The DFT calculation also shows that non-covalent functionalization of BNNTs with aromatic rings can give rise to new impurity states within the band gap of pristine BNNTs, suggesting possible carrier doping of BNNTs via selective adsorption of aromatic rings.


Stereotactic Body Radiation Therapy (SBRT) as Salvage Therapy for Oligorecurrent Pleural Mesothelioma After Multi-Modality Therapy.
Introduction: Therapy options for patients with oligoprogressive malignant pleural mesothelioma (MPM) are limited. Stereotactic Body Radiotherapy (SBRT) may be a promising therapeutic option, as it delivers a localized ablative dose of radiation and therefore balances efficacy and treatment related toxicities. The intent of this retrospective analysis was to evaluate the feasibility of SBRT for limited pleural recurrences. Methods and Materials: This retrospective single-institution study is based on the 21 consecutive patients treated with hypofractionated radiotherapy for oligoprogressive MPM. Clinical and radiological data was collected at regular follow-up visits including toxicity, local control and survival. Results: At primary diagnosis, 57% of the patients presented with stage III disease. Initial treatment of MPM consisted of induction chemotherapy (n = 12) prior to a macroscopic complete resection (n = 18). Three patients received additional intracavitary chemotherapy and another three patients were treated with chemotherapy alone without another treatment at the time of first diagnosis. A total of 50 lesions in recurrent MPM were treated with SBRT. The median number of radiotherapy fractions was 5 (range 3-20) with a median dose per fraction of 5 Gy (range 2.5-12.5 Gy). The median total treatment dose was 30 Gy (20-50 Gy) with a median prescription isodose line (IDL) of 65% (65-100%). Median follow-up of all patients from diagnosis was 28 months (range 7-152 months). Analyzing all lesions separately, the 12-months-local control from SBRT was 73.5%. The median progression free survival (PFS) after SBRT was 6 months (range 0-21 months) and the median OS from first first SBRT was 29 months (range 0-61 months). Only one patients experienced above Grade 3 toxicities. Conclusion: This analysis demonstrates the feasibility of a SBRT approach for oligorecurrent MPM. SBRT was well-tolerated even after multiple repetitions and local control was high with a promising median OS.


Local abundance and regional distribution of tree species of forest fragments in Brazil: a test of models.
Patterns of local abundance and regional distribution of tree species were analyzed on three spatial scales and compared with those predicted by Hanski's 1982 original core-satellite model, Brown's 1984 niche-based model, and Tokeshi's 1992 model. Data were collected in seven forest fragments dispersed over approximately 7200 km2, remnants of a former continuous forest in the south of Mato Grosso do Sul state, southwestern Brazil. Fifty-six east-west-oriented 10 m yen 150 m plots were randomly demarcated in those fragments from 1994 to 1996. In each plot all trees having DBH > or = 0.15 m were recorded and measured. The smallest scale was that of the fragments themselves and the greatest was that of the entire region. No evidence was found of the bimodality predicted by Hanski's 1982 model on any scale. On all scales, an unimodal pattern was found whose mode was on the left side (satellite mode) of the distribution. This pattern was in best agreement with the predictions of Tokeshi's 1992 model.


Molecular biology. Cancer fighter's modus operandi revealed.
Researchers have deciphered how a promising cancer drug acts like a smart bomb, homing in on only a very narrow range of its potential targets in the cell. The compound, known as STI-571, has shown remarkable success in early clinical trials on patients with chronic myelogenous leukemia. Now, in work reported on page 1938, scientists reveal just how the compound works--information that could aid in the design of similar cancer therapies.


21st-Century talent spotting.
How can a person who seems so qualified for a position fail miserably in it? How can someone who clearly lacks relevant skills and experience succeed? The answer is potential, the ability to adapt and grow into increasingly complex roles and environments. For the past several decades, organizations have based their hiring decisions on competencies. But we have entered a new era of talent spotting. Geopolitics, business, industries, and jobs are changing so rapidly that it's impossible to predict the capabilities employees and leaders will need even a few years out. The question now is not whether people have the right skills; it's whether they have the potential to learn new ones. Research points to five markers of potential: a strong motivation to excel in the pursuit of challenging goals combined with the humility to put the group ahead of individual needs; an insatiable curiosity to explore new ideas and avenues; keen insight into connections that others don't see; a strong engagement with work and people; and the determination to overcome obstacles. Once organizations have hired true high potentials--a challenge, given the increasing scarcity of senior talent-and identified the ones they already have, it's crucial to focus on retaining them and on helping them live up to their potential by offering development opportunities that push them out of their comfort zones.


E. coli colonization of the mammalian colon: understanding the process.
Overall, the risk assessment data have shown not only that recombinant DNA research using E. coli strains is safe but also that E. coli strains, in general, including K-12 strains, can colonize the mammalian colon in individuals undergoing antibiotic treatment. However, very little is known about how E. coli colonizes the mammalian colon and it is possible that different strains use different adhesins and different colonic receptors in the process. Through the approach described here it has been possible to begin to identify both E. coli adhesins and colonic receptors which may play an important role in the colonization process. Hopefully, continued research into the molecular basis of E. coli colonic colonization will lead to the development of healthy E. coli strains for recombinant DNA research which are unable to colonize the human colon under any circumstance.


Measuring surgical trainee perceptions to assess the operating room educational environment.
To determine measurable differences in the perception of learning between junior and senior residents in the operating rooms of an obstetrics and gynecology (OBGYN) residency program. Using a cross-sectional design, the Operating Room Educational Environment Measure (OREEM), a 40-item educational environment inventory, was administered to 28 OBGYN residents from 1 training program, who train at 3 hospital sites. The OREEM measures a trainee's perceptions of the teaching surgeon, learning opportunities, operating room atmosphere, and workload. The primary outcome was total OREEM scores and secondary outcomes were OREEM subscale scores, global impression of education, and internal consistency and validity of the OREEM scale. Group sample sizes of 10 and 10 achieved 80% power to detect a 10% difference between group mean OREEM scores +/- 10% with a significance level of 0.05. Twenty-four residents including 11 junior (postgraduate years 1 and 2) and 13 senior (postgraduate years 3 and 4) residents were included in the analysis. Total OREEM scores, learning opportunities, and workload/support subscale scores were significantly lower for junior residents compared with senior residents across all sites. Perceptions of learning at a multispecialty tertiary referral hospital were lower than the community and regional hospitals. This was secondary to complexity of cases, subspecialty fellows, and decreased opportunities to first-assist in the operating room. The OREEM demonstrated acceptable reliability and construct validity. There are measurable differences in perception of the operating room educational environment between junior and senior OBGYN residents using the reliable and valid Operating Room Educational Environment Measure.


Synthesis and in vitro antiproliferative evaluation of d-secooxime derivatives of 13β- and 13α-estrone.
d-Secooximes were synthesized from the d-secoaldehydes in the 13β- and 13α-estrone series. The oximes were modified at three sites in the molecule: the oxime function was transformed into an oxime ether, oxime ester or nitrile group, the propenyl side-chain was saturated and the 3-benzyl ether was removed in order to obtain a phenolic hydroxy function. Triazoles were formed via Cu(I)-catalysed azide-alkyne cycloaddition (CuAAC) from 3-(prop-2-yniloxy)-d-secooximes and benzyl azides. All the products were evaluated in vitro by means of MTT assays for antiproliferative activity against a panel of human adherent cell lines (HeLa, MCF-7, A2780 and A431). Some of them exhibited activities with submicromolar IC50 values, better than that of the reference agent cisplatin. The structural modifications led to significant differences in the cytostatic properties. Flow cytometry indicated that one of the most potent agents resulted in a cell cycle blockade.


Evidence for the inhibition of fertilization in vitro by anti-ZP3 antisera derived from DNA vaccine.
Previously we have found that DNA vaccine, pCMV4-rZPC' can generate specific antibodies against rabbit ZPC (amino acid 263-415, rZPC'), which binds to ovarian ZP and leads to a significant reduction of fertility in vivo. The purpose of this study was to evaluate the effect of antisera from pCMV4-rZPC(')-immunized mice on sperm-oocyte interaction in vitro. The effect of antisera from DNA vaccine-immunized mice on fertilization and early embryonic development was studied using an in vitro fertilization system. The results showed that the antisera supplemented in fertilization medium (10%, v/v) significantly decreased the rate of fertilization compared to that of control groups (P<0.05); whereas the antisera showed no significant effect on the rate of fertilization when ZP-free eggs were used. Moreover, the antisera pre-neutralized with mouse soluble zona pellucida lost the capacity to inhibit fertilization when compared with that of control groups. In addition, the antisera showed no detrimental effect on early developmental potential of mouse embryos in vitro. Taken together, our study provided herein direct evidence showing that antisera generated by DNA vaccine can block sperm-egg recognition during fertilization via targeting the oocyte ZP proteins.


Electrophoretic separations of large DNA molecules by periodic inversion of the electric field.
In gel electrophoresis, nucleic acids and protein-detergent complexes larger than a threshold size all migrate at the same rate. For DNA molecules, this effect can be overcome by the simple procedure of periodically inverting the electric field. Tuning the frequency of the field inversions from 10 to 0.01 hertz, makes it possible to resolve selectively DNA's in the size range 15 to greater than 700 kilobase pairs.


Transformation of Saccharomyces cerevisiae and other fungi: methods and possible underlying mechanism.
Transformation (i.e., genetic modification of a cell by the incorporation of exogenous DNA) is indispensable for manipulating fungi. Here, we review the transformation methods for Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans, Pichia pastoris and Aspergillus species and discuss some common modifications to improve transformation efficiency. We also present a model of the mechanism underlying S. cerevisiae transformation, based on recent reports and the mechanism of transfection in mammalian systems. This model predicts that DNA attaches to the cell wall and enters the cell via endocytotic membrane invagination, although how DNA reaches the nucleus is unknown. Polyethylene glycol is indispensable for successful transformation of intact cells and the attachment of DNA and also possibly acts on the membrane to increase the transformation efficiency. Both lithium acetate and heat shock, which enhance the transformation efficiency of intact cells but not that of spheroplasts, probably help DNA to pass through the cell wall.


The effect of lesions in the locus coeruleus on the physiological responses of the cerebral blood vessels in cats.
The effects of cerebral blood flow (CBF) of lesions placed stereotactically in or near the locus coeruleus were studied in 15 lightly anesthetized cats; 5 control cats in which the electrode was placed but no lesion created, and 10 experimental cats in which a lesion was created. The response of CBF to changes in Paco2 and in mean arterial blood pressure was determined by 133Xe-washout studies 10 days after the stereotactic procedures. The sites of the lesions were studied histologically, and their effects on catecholamine concentrations in the paraventricular hypothalamic nucleus, anterior ventral nucleus of the thalamus, and parietal cortex were determined by radio-chemical assay. Control animals and those with lesions near, but not in, the locus coeruleus had normal Paco2--CBF response curves and normal catecholamine concentrations in the areas of biopsy. Bilateral destruction of the locus coeruleus was confirmed in 3 animals on histological examination and in these animals there were decreased levels of catecholamines in the areas of assay, higher resting CBFs at normocapnia, and significantly abnormal CBF--Paco2 response curves. The autoregulatory response to changes in perfusion pressure was preserved. Thus, noradrenergic neurons originating in the locus coeruleus may contribute to the control of intraparenchymal cerebral vessels and disturbance of this control may be important in the pathology of cerebral ischemia.


[Use of dimephosphon in pathogenetic therapy of experimental tuberculosis].
The results of a study of a new synthetic drug dimephosphon used as a pathogenetic means in the treatment of experimental tuberculosis are presented. Dimephosphon was found to be responsible for both the in vivo and in vitro decrease of the degree of MBT resistance to rifampicin. The findings of macroscopic, histologic and bacteriologic examinations demonstrated a significant increase in the effectiveness of antituberculous therapy. Dimephosphon monotherapy in mice elicited manifested stimulation of peritoneal macrophages: increase in O2- production, and decline in extracellular 5-nucleotidase activity. Nemolysine-synthetic cellular splenic activity in mice rose essentially. No direct stimulating influence of dimephosphon on functional macrophage activity in vitro was found.


Single cholinergic receptor channel currents in cultured human muscle.
Single cholinergic channel currents were recorded in adult human muscle tissue culture. The agonists suberyldicholine and carbamylcholine produce channels with the same conductance as channels produced by acetylcholine but with different closing kinetics. The antagonist tubocurarine, alone or mixed with suberyldicholine, activates channels which close very rapidly. For agonist-activated channels, the distribution of open state lifetimes shows deviations from the usual single exponential form. An excess of short duration openings indicates the presence of an additional faster kinetic process. The lifetime distribution data can be interpreted in terms of varying proportions of slow and fast components which are present in a ratio determined by curve-fitting the appropriate two-exponential function to observed open time distributions. This ratio shows great variability in muscle from older cultures, but the fast and slow time constants are relatively constant. The observation of double exponential open time distributions indicates that the mechanism of channel closing is more complicated than earlier evidence indicated.


Volume and diagnosis: an approach to cross-border care in eight European countries.
Mobility of patients is a pertinent issue on the European Union's agenda. This study aimed to estimate the volume and main diagnoses of cross-border care in eight European countries, in order to provide policy makers with background information about the nature of patient mobility in Europe. This article reports the combined findings from three independent studies that compiled self-reported information on admissions data and main diagnoses from more than 200 hospitals in eight European countries. The average volume of cross-border patients accounted for less than 1% of total admissions in the hospitals studied here. Diseases of the circulatory system (mainly acute myocardial infarction) and fractures were the most common reasons for hospitalisation of European patients abroad. Deliveries and other diagnoses related to pregnancy, pneumonia, appendicitis and other diseases of the digestive system, aftercare procedures, and disorders of the eye and adnexa were also common diagnoses for this population. Hospitals should reinforce their efforts to adapt the care provided to the needs of foreign patients in treatment areas that cover the most frequent pathologies identified in this population.


Mixed Salmonella infection: a report of two cases from the Aga Khan University Hospital, Karachi.
Enteric fever remains a major health problem in the developing world, including Pakistan. Poor sanitation and hygienic conditions are the major predisposing factors. Salmonella infection with different strains in the same patient has rarely been reported previously. We are reporting two cases of bacteraemia with simultaneous detection of two strains of Salmonella in a single episode of infection. In both the cases, 2 different serotypes of Salmonella were causing bacteraemia leading to fever. In highly endemic area, one must be aware of mixed Salmonella infections as inappropriate diagnosis of such infections may lead to treatment failure.


Cardiac Safety of Dual Anti-HER2 Therapy in the Neoadjuvant Setting for Treatment of HER2-Positive Breast Cancer.
Trastuzumab and pertuzumab are approved for the neoadjuvant treatment of human epidermal growth receptor 2 (HER2)-positive breast cancer, but cardiac safety data is limited. We report the cardiac safety of dose-dense doxorubicin and cyclophosphamide (AC) followed by paclitaxel, trastuzumab, and pertuzumab (THP) in the neoadjuvant setting followed by adjuvant trastuzumab-based therapy. Fifty-seven patients treated with neoadjuvant dose-dense AC-THP followed by adjuvant trastuzumab-based therapy between September 1, 2013, and March 1, 2015, were identified. The primary outcome was cardiac event rate, defined by heart failure (New York Heart Association [NYHA] class III/IV) or cardiac death. Patients underwent left ventricular ejection fraction (LVEF) monitoring at baseline, after AC, and serially during 1 year of anti-HER2 therapy. The median age was 46 years (range 26-68). Two (3.5%) patients developed NYHA class III/IV heart failure 5 and 9 months after initiation of trastuzumab-based therapy, leading to permanent discontinuation of anti-HER2 treatment. Seven (12.3%) patients developed a significant LVEF decline (without NYHA class III/IV symptoms). The median LVEF was 65% (range 55%-75%) at baseline and 64% (range 53%-72%) after AC, and decreased to 60% (range 35%-70%), 60% (range 23%-73%), 61% (range 25%-73%), and 58% (range 28%-66%) after 3, 6, 9, and 12 months (± 6 weeks) of trastuzumab-based therapy. The incidence of NYHA class III/IV heart failure after neoadjuvant AC-THP (followed by adjuvant trastuzumab-based therapy) is comparable to rates reported in trials of sequential doxorubicin and trastuzumab. Our findings do not suggest an increased risk of cardiotoxicity from trastuzumab plus pertuzumab following a doxorubicin-based regimen. Dual anti-human epidermal growth receptor 2 (HER2) therapy with trastuzumab and pertuzumab combined with standard chemotherapy has received accelerated approval for the neoadjuvant treatment of stage II-III HER2-positive breast cancer. Cardiac safety data for trastuzumab and pertuzumab in this setting are limited to clinical trials that utilized epirubicin-based chemotherapy. Formalized investigations into the cardiac safety of trastuzumab and pertuzumab with doxorubicin- (rather than epirubicin) based regimens are important because these regimens are widely used for the adjuvant and neoadjuvant treatment of breast cancer. The known role of HER2 signaling in the physiological adaptive responses of the heart provides further rationale for study on the potential cardiotoxicity of dual anti-HER2 blockade. Findings from this retrospective study provide favorable preliminary data on the cardiac safety of trastuzumab and pertuzumab in combination with a regimen of neoadjuvant doxorubicin and cyclophosphamide followed by paclitaxel, one of the preferred breast cancer treatment regimens, according to the National Comprehensive Cancer Network.


Rearing density and susceptibility of Rana pipiens metamorphs to cercariae of a digenetic trematode.
Cercariae of many digenean trematodes target particular developmental stages of their hosts. For some digeneans that are parasites of amphibians, infection appears timed to host metamorphosis. The success and timing of metamorphosis is itself affected by a number of factors, including host density. We investigated the degree to which rearing density of Rana pipiens larvae influenced time to metamorphosis and snout-to-vent length and mass at metamorphosis, as well as establishment of cercariae of the trematode Manodistomum syntomentera Stafford, 1905. As expected, individuals metamorphosed later, were smaller, and weighed less at metamorphosis if they were reared under intermediate to high densities compared with low densities, in experimental outdoor mesocosms. Cercariae establishment was higher in smaller metamorphs that took longer to metamorphose within the low-density treatment. Additionally, cercariae establishment was lower in larvae from the low-density tanks compared with larvae from the intermediate- to high-density tanks. However, more tadpoles had failed to metamorphose in the intermediate to high rearing densities by the time cercariae were no longer available from natural collections of first intermediate hosts, Physa spp. Larval amphibians under crowded conditions should experience increased susceptibility to trematode establishment in nature, but only if they metamorphose within the time period when cercariae are still available.


Ferroportin Q248h, dietary iron, and serum ferritin in community African-Americans with low to high alcohol consumption.
Alcohol consumption is associated with increased iron stores. In sub-Saharan Africa, high dietary ionic iron and the ferroportin Q248H allele have also been implicated in iron accumulation. We examined the associations of ferroportin Q248H, alcohol and dietary iron with serum ferritin, aspartate aminotransaminase (AST) and alanine aminotransaminase (ALT) concentrations in African-Americans. Inner-city African-Americans (103 men, 40 women) were recruited from the community according to reported ingestion of >4 alcoholic drinks/d or <2/wk. Typical daily heme iron, nonheme iron and alcohol were estimated using University of Hawaii's multiethnic dietary questionnaire. Based on dietary questionnaire estimates we established categories of < versus > or =56 g alcohol/d, equivalent to 4 alcoholic drinks/d assuming 14 g alcohol per drink. Among 143 participants, 77% drank <56 g alcohol/d and 23%> or =56 g/d as estimated by the questionnaire. The prevalence of ferroportin Q248H was 23.3% with alcohol >56 g/d versus 7.5% with lower amounts (p = 0.014). Among subjects with no history of HIV disease, serum ferritin concentration had positive relationships with male gender (p = 0.041), alcohol consumption (p = 0.021) and ALT concentration (p = 0.0001) but not with dietary iron intake or ferroportin Q248H. Serum AST and ALT concentrations had significant positive associations with male gender and hepatitis C seropositivity but not with alcohol or dietary iron intake or ferroportin Q248H. Our findings suggest a higher prevalence of ferroportin Q248H with greater alcohol consumption, and this higher prevalence raises the possibility that the allele might ameliorate the toxicity of alcohol. Our results suggest that alcohol but not dietary iron contributes to higher body iron stores in African-Americans. Studies with larger numbers of participants are needed to further clarify the relationship of ferroportin Q248H with the toxicity of alcohol consumption.


RRB1 and RRB2 encode maize retinoblastoma-related proteins that interact with a plant D-type cyclin and geminivirus replication protein.
Unlike mammalian and yeast cells, little is known about how plants regulate G1 progression and entry into the S phase of the cell cycle. In mammalian cells, a key regulator of this process is the retinoblastoma tumor suppressor protein (RB). In contrast, G1 control in Saccharomyces cerevisiae does not utilize an RB-like protein. We report here the cloning of cDNAs from two Zea mays genes, RRB1 and RRB2, that encode RB-related proteins. Further, RRB2 transcripts are alternatively spliced to yield two proteins with different C termini. At least one RRB gene is expressed in all the tissues examined, with the highest levels seen in the shoot apex. RRB1 is a 96-kDa nuclear protein that can physically interact with two mammalian DNA tumor virus oncoproteins, simian virus 40 large-T antigen and adenovirus E1A, and with a plant D-type cyclin. These associations are abolished by mutation of a conserved cysteine residue in RRB1 that is also essential for RB function. RRB1 binding potential is also sensitive to deletions in the conserved A and B domains, although differences exist in these effects compared to those of human RB. RRB1 can also bind to the AL1 protein from tomato golden mosaic virus (TGMV), a protein which is essential for TGMV DNA replication. These results suggest that G1 regulation in plant cells is controlled by a mechanism which is much more similar to that found in mammalian cells than that in yeast.


Higher Fat Content in Breastmilk Expressed Manually: A Randomized Trial.
This study aimed to evaluate the effect of milk expression method (manual expression versus electric pump) on the composition of breastmilk. Data on 21 mothers of 21 newborns 48-72 hours postdelivery were collected and analyzed. The women were randomly assigned to express breastmilk manually followed by pump, or in reverse order. The fat, carbohydrate, and protein contents of the milk samples were analyzed using a human milk analyzer (Miris AB, Uppsala, Sweden). The fat and energy contents of milk obtained through manual expression were higher than those obtained by pump (p=0.024 and p=0.04, respectively, by the Wilcoxon signed rank test). There were no significant differences in protein or carbohydrate content of milk obtained by either method of expression. The difference in fat content between milk obtained by the two methods was not correlated with mother's age, delivery method, gestational age at delivery, parity, or the interval between delivery and the time the sampled milk was obtained. Manually expressed human milk had higher fat content than milk expressed by electric pump. We speculate that this difference is due to the presence of hindmilk in the manually expressed milk because the technique of massaging the breast during manual expression is more likely than the pump to eject hindmilk, which has been shown to have higher fat content than foremilk.


Barriers to accessing HIV/AIDS care in North Carolina: rural and urban differences.
Many HIV-positive individuals face multiple barriers to care and therefore frequently experience unmet medical and support services needs. Rural areas often lack the infrastructure to support the delivery of comprehensive HIV services; however, few studies have examined service barriers faced by rural residents with HIV/AIDS, particularly in the South where two-thirds of people living with HIV/AIDS in rural areas reside. We surveyed North Carolina HIV/AIDS case managers (N = 111) employed at state-certified agencies regarding barriers to medical and support services that influence medication adherence for their rural and urban-living clients. For each of the seven barriers assessed (long travel for care, HIV-related stigma, and a lack of transportation; HIV-trained medical practitioners; housing; mental health services and substance abuse treatment), a substantial proportion of case managers (29-67%) reported it was a 'major problem'. For five of the seven barriers, rural case managers were significantly more likely to identify the barrier as a 'major problem'. Multivariate analysis revealed that rural case managers and case managers with more female clients reported a greater number of barriers. Because unmet medical and support service needs may result in poorer outcomes for HIV-positive individuals, barriers to these services must be identified and addressed, particularly in rural areas which may be highly underserved.


[Variant anatomy of maxillar sinus according to the data of computer tomography with 3D function].
The paper presents the data on variant and individual anatomy of the maxillary sinus in 400 men and women of the I and II periods of adulthood obtained using by computer tomography with 3D function. It was found that in the I period of adulthood, 12.5% of individuals had the bony septa in the alveolar recess of the maxillary sinus which were oriented frontally--Underwood's septa. Individuals of the II period of adulthood had this septa in 20% of cases. Also in the second age group, 7.2% of the cases demonstrated the septa which are oriented sagittally. These structures were often associated with the bony canal of the infraorbital nerve. The detected variants of individual variability of the maxillary sinus have both theoretical and practical importance. In particular, during the planning of micromaxillotomy (through the endonasal access or fossa canina), the presence of bone septa, their location and the course of the infraorbital nerve canal should be taken into account.


Effect of medetomidine administration on bispectral index measurements in dogs during anesthesia with isoflurane.
To determine the relationship between bispectral index (BIS) and minimum alveolar concentration (MAC) multiples of isoflurane after IM injection of medetomidine or saline (0.9% NaCl) solution in anesthetized dogs. 6 dogs. Each dog was anesthetized 3 times with isoflurane. First, the MAC of isoflurane for each dog was determined by use of the tail clamp method. Second, anesthetized dogs were randomly assigned to receive an IM injection of medetomidine (8 microg x kg(-1)) or an equal volume of isotonic saline (0.9% NaCl) solution 30 minutes prior to beginning BIS measurements. Last, anesthetized dogs received the remaining treatment (medetomidine or isotonic saline solution). Dogs were anesthetized at each of 4 MAC multiples of isoflurane. Ventilation was controlled and atracurium (0.2 mg/kg followed by 6 microg/kg/min as a continuous infusion, IV) administered. After a 20-minute equilibration period at each MAC multiple of isoflurane, BIS data were collected for 5 minutes and median values of BIS calculated. BIS significantly decreased with increasing MAC multiples of isoflurane over the range of 0.8 to 2.0 MAC. Mean (+/- SD) MAC of isoflurane was 1.3 +/- 0.2%. During isoflurane-saline anesthesia, mean BIS measurements at 0.8, 1.0, 1.5, and 2.0 MAC were 65 +/- 8, 60 +/- 7 52 +/- 3, and 31 +/- 28, respectively. During isoflurane-medetomidine anesthesia, mean BIS measurements at 0.8, 1.0, 1.5, and 2.0 MAC were 77 +/- 4, 53 +/- 7, 31 +/- 24, and 9 +/- 20, respectively. BIS monitoring in dogs anesthetized with isoflurane has a predictive value in regard to degree of CNS depression. During isoflurane anesthesia, our results support a MAC-reducing effect of medetomidine.


Initial experience with intraoperative magnetic resonance imaging in spine surgery.
A case series of 12 patients who underwent spine surgery in an intraoperative magnetic resonance imager (IMRI). To determine the advantages, limitations, and potential applications to spine surgery of the IMRI. Existing stereotactic navigational systems are limited because images are obtained before surgery and are not updated to reflect intraoperative changes. In addition, they necessitate manual registration of fiducial landmarks on the patient's anatomy by the surgeon to the previously obtained image data set, which is a potential source of error. The IMRI eliminates these difficulties by using intraoperative acquisition of MRI images for surgical navigation with the capacity for both image update and image-guided frameless stereotaxy. The IMRI is a novel cryogenless superconducting magnet with an open configuration that allows the surgeon full access to the patient during surgery and intraoperative imaging. T1- and T2-weighted fast spin echo images were obtained for localization, after surgical exposure and after decompression during the course of 12 spine surgeries performed in the IMRI. The authors performed a series of 12 procedures in the IMRI that included three lumbar discectomies, three anterior cervical discectomies with allograft fusion, three cervical vertebrectomies with allograft fusion, two cervical foraminotomies, and one decompressive cervical laminectomy. The system provided rapid and accurate localization in all cases. The adequacy of decompression by MRI during surgery was confirmed in 10 of 12 cases. The IMRI provided accurate and rapid localization in all cases and confirmed the adequacy of decompression in the majority of cases. Future applications of the IMRI to spine surgery may include intraoperative guidance for resection of spine and spinal cord tumors and trajectory planning for spinal endoscopy or screw fixation.