A novel chimpanzee adenovirus vector with low human seroprevalence: improved systems for vector derivation and comparative immunogenicity.
Recombinant adenoviruses are among the most promising tools for vaccine antigen delivery. Recently, the development of new vectors has focused on serotypes to which the human population is less exposed in order to circumvent pre-existing anti vector immunity. This study describes the derivation of a new vaccine vector based on a chimpanzee adenovirus, Y25, together with a comparative assessment of its potential to elicit transgene product specific immune responses in mice. The vector was constructed in a bacterial artificial chromosome to facilitate genetic manipulation of genomic clones. In order to conduct a fair head-to-head immunological comparison of multiple adenoviral vectors, we optimised a method for accurate determination of infectious titre, since this parameter exhibits profound natural variability and can confound immunogenicity studies when doses are based on viral particle estimation. Cellular immunogenicity of recombinant E1 E3-deleted vector ChAdY25 was comparable to that of other species E derived chimpanzee adenovirus vectors including ChAd63, the first simian adenovirus vector to enter clinical trials in humans. Furthermore, the prevalence of virus neutralizing antibodies (titre >1:200) against ChAdY25 in serum samples collected from two human populations in the UK and Gambia was particularly low compared to published data for other chimpanzee adenoviruses. These findings support the continued development of new chimpanzee adenovirus vectors, including ChAdY25, for clinical use.


Aldosterone abrogates nuclear factor kappaB-mediated tumor necrosis factor alpha production in human neutrophils via the mineralocorticoid receptor.
Mineralocorticoid receptor (MR) activation by aldosterone controls salt homeostasis and inflammation in several tissues and cell types. Whether or not a functional MR exists in polymorphonuclear neutrophils is unknown. We investigated the hypothesis that aldosterone modulates inflammatory neutrophil responses via the MR. By flow cytometry, Western blot analysis, and microscopy, we found that neutrophils possess MR. Preincubation with aldosterone (10(-11) to 10(-6) M) dose-dependently inhibited nuclear factor kappaB activation in interleukin (IL)-8- and granulocyte/macrophage colony-stimulating factor-treated neutrophils on fibronectin by IkappaBalpha Western blotting, electrophoretic mobility shift assay, and RT-PCR for IkappaBalpha mRNA. Aldosterone had no effect on tumor necrosis factor alpha- and lipopolysaccharide-mediated nuclear factor kappaB activation or on IL-8- and granulocyte/macrophage colony-stimulating factor-induced extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, or phosphatidylinositol 3-kinase/Akt activation. Spironolactone prevented nuclear factor kappaB inhibition, indicating an MR-specific aldosterone effect. By RT-PCR, we found that neutrophils have 11beta-hydroxysteroid dehydrogenase. Tumor necrosis factor alpha, which is controlled by nuclear factor kappaB, increased in the cell supernatant with IL-8 treatment. Aldosterone completely prevented this effect. RT-PCR showed a strong tumor necrosis factor alpha mRNA increase with IL-8 that was blocked by aldosterone, excluding the possibility that the tumor necrosis factor alpha increase was merely a consequence of secretion. Finally, conditioned medium from IL-8-treated neutrophils increased intercellular adhesion molecule-1 expression on endothelial cells and subsequently the adhesion of IL-8-treated neutrophils to endothelial cells. These effects were reduced when conditioned medium from aldosterone-pretreated neutrophils was used, and spironolactone blocked the aldosterone effect. Our data indicate that a functional MR exists in neutrophils mediating antiinflammatory effects that are at work when neutrophils interact with endothelial cells. These data could be relevant to MR-blockade treatment protocols.


Typhoid Outbreaks, 1989-2018: Implications for Prevention and Control.
Typhoid fever remains an important public health problem in low- and middle-income countries, with large outbreaks reported from Africa and Asia. Although the WHO recommends typhoid vaccination for control of confirmed outbreaks, there are limited data on the epidemiologic characteristics of outbreaks to inform vaccine use in outbreak settings. We conducted a literature review for typhoid outbreaks published since 1990. We found 47 publications describing 45,215 cases in outbreaks occurring in 25 countries from 1989 through 2018. Outbreak characteristics varied considerably by WHO region, with median outbreak size ranging from 12 to 1,101 cases, median duration from 23 to 140 days, and median case fatality ratio from 0% to 1%. The largest number of outbreaks occurred in WHO Southeast Asia, 13 (28%), and African regions, 12 (26%). Among 43 outbreaks reporting a mode of disease transmission, 24 (56%) were waterborne, 17 (40%) were foodborne, and two (5%) were by direct contact transmission. Among the 34 outbreaks with antimicrobial resistance data, 11 (32%) reported Typhi non-susceptible to ciprofloxacin, 16 (47%) reported multidrug-resistant (MDR) strains, and one reported extensively drug-resistant strains. Our review showed a longer median duration of outbreaks caused by MDR strains (148 days versus 34 days for susceptible strains), although this difference was not statistically significant. Control strategies focused on water, sanitation, and food safety, with vaccine use described in only six (13%) outbreaks. As typhoid conjugate vaccines become more widely used, their potential role and impact in outbreak control warrant further evaluation.


Nationwide risk analysis of duodenoscope and linear echoendoscope contamination.
Contaminated duodenoscopes and linear echoendoscopes (DLEs) pose a risk for infectious outbreaks. To identify DLEs and reprocessing risk factors, we combined the data from the previously published nationwide cross-sectional PROCESS 1 study (Prevalence of contamination of complex endoscopes in the Netherlands) with the follow-up PROCESS 2 study. We invited all 74 Dutch DLE centers to sample ≥2 duodenoscopes during PROCESS 1, and all duodenoscopes as well as linear echoendoscopes during PROCESS 2. The studies took place 1 year after another. Local staff sampled each DLE at ≤6 sites according to uniform methods explained by online videos. We used 2 contamination definitions: (1) any microorganism with ≥20 colony-forming units (CFU)/20 mL (AM20) and (2) presence of microorganisms with GI or oral origin, independent of CFU count (MGOs). We assessed the factors of age and usage by performing an analysis of pooled data of both PROCESS studies; additional factors including reprocessing characteristics were only recorded in PROCESS 2. Ninety-seven percent of all Dutch centers (72 of 74; PROCESS 1, 66; PROCESS 2, 61) participated in one of the studies, sampling 309 duodenoscopes and 64 linear echoendoscopes. In total, 54 (17%) duodenoscopes and 8 (13%) linear echoendoscopes were contaminated according to the AM20 definition. MGOs were detected on 47 (15%) duodenoscopes and 9 (14%) linear echoendoscopes. Contamination was not age or usage dependent (all P values ≥.27) and was not shown to differ between the reprocessing characteristics (all P values ≥.01). In these nationwide studies, we found that DLE contamination was independent of age and usage. These results suggest that old and heavily used DLEs, if maintained correctly, have a similar risk for contamination as new DLEs. The prevalence of MGO contamination of ∼15% was similarly high for duodenoscopes as for linear echoendoscopes, rendering patients undergoing ERCP and EUS at risk for transmission of microorganisms.


Overview of the protein-protein interaction annotation extraction task of BioCreative II.
The biomedical literature is the primary information source for manual protein-protein interaction annotations. Text-mining systems have been implemented to extract binary protein interactions from articles, but a comprehensive comparison between the different techniques as well as with manual curation was missing. We designed a community challenge, the BioCreative II protein-protein interaction (PPI) task, based on the main steps of a manual protein interaction annotation workflow. It was structured into four distinct subtasks related to: (a) detection of protein interaction-relevant articles; (b) extraction and normalization of protein interaction pairs; (c) retrieval of the interaction detection methods used; and (d) retrieval of actual text passages that provide evidence for protein interactions. A total of 26 teams submitted runs for at least one of the proposed subtasks. In the interaction article detection subtask, the top scoring team reached an F-score of 0.78. In the interaction pair extraction and mapping to SwissProt, a precision of 0.37 (with recall of 0.33) was obtained. For associating articles with an experimental interaction detection method, an F-score of 0.65 was achieved. As for the retrieval of the PPI passages best summarizing a given protein interaction in full-text articles, 19% of the submissions returned by one of the runs corresponded to curator-selected sentences. Curators extracted only the passages that best summarized a given interaction, implying that many of the automatically extracted ones could contain interaction information but did not correspond to the most informative sentences. The BioCreative II PPI task is the first attempt to compare the performance of text-mining tools specific for each of the basic steps of the PPI extraction pipeline. The challenges identified range from problems in full-text format conversion of articles to difficulties in detecting interactor protein pairs and then linking them to their database records. Some limitations were also encountered when using a single (and possibly incomplete) reference database for protein normalization or when limiting search for interactor proteins to co-occurrence within a single sentence, when a mention might span neighboring sentences. Finally, distinguishing between novel, experimentally verified interactions (annotation relevant) and previously known interactions adds additional complexity to these tasks.


Protein targets of thioacetamide metabolites in rat hepatocytes.
Thioacetamide (TA) has long been known as a hepatotoxicant whose bioactivation requires S-oxidation to thioacetamide S-oxide (TASO) and then to the very reactive S,S-dioxide (TASO2). The latter can tautomerize to form acylating species capable of covalently modifying cellular nucleophiles including phosphatidylethanolamine (PE) lipids and protein lysine side chains. Isolated hepatocytes efficiently oxidize TA to TASO but experience little covalent binding or cytotoxicity because TA is a very potent inhibitor of the oxidation of TASO to TASO2. However, hepatocytes treated with TASO show extensive covalent binding to both lipids and proteins accompanied by extensive cytotoxicity. In this work, we treated rat hepatocytes with [(14)C]-TASO and submitted the mitochondrial, microsomal, and cytosolic fractions to 2DGE, which revealed a total of 321 radioactive protein spots. To facilitate the identification of target proteins and adducted peptides, we also treated cells with a mixture of TASO/[(13)C2D3]-TASO. Using a combination of 1DGE- and 2DGE-based proteomic approaches, we identified 187 modified peptides (174 acetylated, 50 acetimidoylated, and 37 in both forms) from a total of 88 nonredundant target proteins. Among the latter, 57 are also known targets of at least one other hepatotoxin. The formation of both amide- and amidine-type adducts to protein lysine side chains is in contrast to the exclusive formation of amidine-type adducts with PE phospholipids. Thiobenzamide (TB) undergoes the same two-step oxidative bioactivation as TA, and it also gives rise to both amide and amidine adducts on protein lysine side chains but only amidine adducts to PE lipids. Despite their similarity in functional group chemical reactivity, only 38 of 62 known TB target proteins are found among the 88 known targets of TASO. The potential roles of protein modification by TASO in triggering cytotoxicity are discussed in terms of enzyme inhibition, protein folding, and chaperone function, and the emerging role of protein acetylation in intracellular signaling and the regulation of biochemical pathways.


Stimulated neutrophils induce myosin light chain phosphorylation and isometric tension in endothelial cells.
The mechanism or mechanisms by which polymorphonuclear leukocytes (PMN) penetrate junctions between neighboring endothelial cells (EC) to traverse endothelial barriers remain unresolved. We report that chemoattractant-stimulated PMN induce a coordinate increase in both phosphorylation of serine 19 and threonine 18 of EC myosin regulatory light chains and isometric tension generation by EC monolayers. Unstimulated PMN had no effect on either parameter. These findings, coupled with our previous report (Huang et al., J. Cell Biol. 120: 1371-1380, 1993) that chemoattractant-stimulated PMN cause a rise in EC cytosolic free Ca2+, provide strong presumptive evidence that myosin light chain kinase is the EC enzyme responsible for initiating myosin light chain phosphorylation, EC contraction, and isometric tension generation in response to chemoattractant-stimulated PMN. We suggest that, by inducing phosphorylation of EC cytoskeletal proteins, chemoattractant-stimulated PMN induce EC to open their intercellular junctions, thereby facilitating transendothelial movement of these leukocytes.


catena-Poly[[(tetra-hydro-furan-κO)potas-sium]-di-μ-dimethyl-amido-κ(4) N:N-aluminium-di-μ-dimethyl-amido-κ(4) N:N-potassium-di-μ-dimethyl-amido-κ(4) N:N-aluminium-di-μ-dimethyl-amido-κ(4) N:N].
The title compound, [Al2K2(C2H6N)8(C4H8O)] n , formed during the sonochemical reaction between Al(NMe2)3 and sodium-potassium alloy in the presence of tetra-hydro-furan (THF). Its asymmetric unit has two inequivalent K(+) sites. One site is coordinated by a THF ligand, and crystallizes as a one-dimensional polymer with a backbone of catenated AlN2K rings. A twofold rotation axis bis-ects one K(+) site and the THF ligand; the second K(+) site is situated on an inversion centre, resulting in a planar four-coordination by N atoms. The latter symmetry operation generates the second half of the THF mol-ecule and fills out the coordination sphere of the potassium sites. The chains extend along the c-axis direction and zigzag at the THF-coordinated K(+) sites by an angle of 106.02 (5)°.


Women behind bars: health needs of inmates in a county jail.
This study was performed to assess the prevalence of behavioral risk factors and correlates of poor self-reported health among incarcerated women in a county jail in Oregon. The authors collected self-reported data from recently incarcerated women at a county jail, focusing on prevalence of high-risk health behaviors, history of health care use, history of physical and sexual abuse, and health care coverage. The authors assessed factors associated with poor self-reported health using logistic regression techniques. More than half of the participants reported a history of intravenous drug use, 67% reported a history of sexual abuse, 79% reported a history of physical abuse, and 43% stated that they had a history of trading sex for money or drugs. Two factors were associated with poor self-reported health: history of physical assault (odds ratio [OR] = 2.7; 95% confidence interval [CI] 1.4, 5.2) and use of heroin during the month prior to arrest (OR = 2.9; 95% CI 1.3, 6.6). The high prevalence of health risk behaviors among the inmates suggests a number of areas for intervention. These findings may also be used to guide topics addressed during intake interviews of new inmates, and to help identify inmates that require additional medical or social services.


Interlimb differences of directional biases for stroke production.
Directional preferences during center-out horizontal shoulder-elbow movements were previously characterized for the dominant arm. These preferences were attributed to a tendency to actively accelerate one joint, while exploiting largely passive motion at the other joint. Since the non-dominant arm is known for inefficient coordination of inter-segmental dynamics, here we hypothesized that directional preferences would differ between the arms. A center-out free-stroke drawing task was used that allowed freedom in the selection of movement directions. The task was performed both with and without a secondary cognitive task that has been shown to increase directional biases of the dominant arm. Mirror-symmetrical directional preferences were observed in both arms, with similar bias strength and secondary task effects. The preferred directions were characterized by maximal exploitation of interaction torques for movement production, but only in the dominant arm. The non-dominant arm failed to benefit from interaction torques. The results point to a hierarchical architecture of control. At the higher level, a movement capable to perform the task while satisfying preferences in joint control is specified through forward dynamic transformations. This process is mediated for both arms from a common neural network adapted to the dominant arm and, specifically, to its ability to exploit interaction torques. Dynamic transformations that determine actual control commands are specified at the lower level of control. An alternative interpretation that strokes might be planned evenly across directions, and biases emerge during movement execution due to anisotropic resistance of intrinsic factors that do not depend on arm dominance is also discussed.


The effect of endoscopic olfactory cleft polyp removal on olfaction.
The presence of olfactory cleft polyps in chronic rhinosinusitis with nasal polyposis is well documented, but the effect of endoscopic olfactory cleft polyp surgery on olfaction, versus observation, has not been well studied. This analysis assessed if microdebridement of olfactory cleft polyps yields significant objective smell improvements in those with anosmia or hyposmia. A randomized prospective single-blinded study was performed on patients undergoing bilateral endoscopic sinus surgery with profound bilateral nasal polyposis, excluding those younger than 18 years or without olfactory polyps. A preoperative University of Pennsylvania Smell Identification Test (UPSIT), visual analog scale (VAS), and sinonasal outcomes 20 score (SNOT-20), and a follow-up at 6 months was performed. Two cohorts were created, including one with cleft polyp removal (group A) and one with cleft polyps left in place (group B). There were 10 patients in group A and 7 in group B. By using the Wilcoxon signed rank test, the two groups were individually analyzed for changes in the preoperative UPSIT, VAS, and SNOT-20 versus the 6-month test results. In group A, the improvement in the UPSIT, VAS, and SNOT-20 were statistically significant at p < 0.05. For group B only the improvement in the VAS was statistically significant, at p < 0.05. There was a statistically significant difference in clinical smell improvement between group A and B at 6 months (p = 0.00512). Evidence exists that olfactory cleft polyp surgery improves olfactory function outcomes. Long-term data beyond 6 months is needed to further validate these early promising outcomes.


Contribution of direct and indirect recognition pathways to T cell alloreactivity.
T cells from an HLA-DR11/DR12 responder were stimulated in mixed lymphocyte culture with cells carrying the DR1 antigen. After priming, T cells proliferated in response to both DR1-positive-stimulating cells and a peptide derived from a polymorphic region of the HLA-DR beta 1*0101 chain presented by responder's antigen-presenting cells (APC). The dominant epitope recognized by the primed T cells corresponded to residue 21-42 and was presented by the responder's HLA-DR12 antigen. The DR1 peptide-reactive T cells express T cell receptor V beta 3. The results demonstrate that allopeptides derived from the processing and presentation of donor major histocompatibility complex molecules by host-derived APC trigger alloreactivity. The frequency of T cells engaged in the indirect pathway of allorecognition is about 100-fold lower than that of T cells participating in the direct recognition of native HLA-DR antigen. However, indirect allorecognition may play an important role in chronic allograft rejection, a phenomenon that is mediated by the activation of T helper cells and of alloantibody-producing B cells.


The effect of information technology investment on firm-level performance in the health care industry.
The return on investment for information technology (IT) has been the subject of much debate throughout the history of management information systems research. Often referred to as the productivity paradox, increased IT investments have not been consistently associated with increased productivity. Understanding individual IT factors that directly contribute to business value should provide insight into the productivity paradox. The effects of 3 different firm-level IT characteristics on financial performance in the health care industry are studied. Specifically, the effects of IT budget, IT outsourcing, and the relative number of IT personnel on firm-level financial performance are analyzed. Regression analysis of archival survey data for 914 Integrated Healthcare Delivery Systems is performed. IT budgetary expenditures and the number of IT services outsourced are associated with increases in the profitability of Integrated Healthcare Delivery Systems, whereas increases in IT personnel are not significantly associated with increased profitability. Each one tenth of a percentage increase in IT expenditures is associated with approximately $100,000 in increased profit, and each additional IT service outsourced is associated with approximately $950,000 in increased profit for an average-sized Integrated Healthcare Delivery System. To increase profitability, IT administrators should increase IT budgetary expenditures along with IT outsourcing levels. IT administrators in the health care industry can use such findings during budgeting cycles to justify increased investments in IT personnel as being budget neutral while increasing organizational capacity.


Population Genetic Structures of Puccinia striiformis in Ningxia and Gansu Provinces of China.
Wheat stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici, is one of the most destructive plant diseases in China. Gansu and Ningxia Provinces are considered to be the key areas for over-summering of this pathogen in China. In this study, 283 P. striiformis f. sp. tritici isolates were collected in these two provinces. Amplified fragment length polymorphism (AFLP) was used to analyze the population genetics and to infer the chance of population exchanges between different geographic locations and seasons. The genotypic diversity of the Gansu population (0.514) was slightly higher than that of the Ningxia population (0.489). The occurrence of frequent population exchanges between these two regions was observed, showing that 40 AFLP genotypes were shared by the populations of the two provinces. Gene flow between these two regions in autumn and spring subpopulations was also detected. The genotype distribution in three populations of Ningxia from opposite sides of the Liupan Mountains revealed possible significant effects of the mountains on limiting gene flow and population exchange. Phylogenetic analysis confirmed the possibility of recombination in some of the studied subpopulations in both provinces.


Oligo(ethylene glycol)-based thermosensitive dendrimers and their tumor accumulation and penetration.
Dendrimers have several featured advantages over other nanomaterials as drug carriers, such as well-defined structure, specific low-nanometer size, and abundant peripheral derivable groups, etc. However, these advantages have not been fully exploited yet to optimize their biological performance, especially tumor penetration, which is a shortcoming of current nanomaterials. Here we show the syntheses of a new class of oligo(ethylene glycol) (OEG)-based thermosensitive dendrimers up to the fourth generation. Each dendrimer shows monodisperse structure. OEG/poly(ethylene glycol) (PEG) moieties with different precise lengths were introduced to the periphery of the fourth-generation dendrimer followed by an antitumor agent, gemcitabine (GEM). The biodistributions of the GEM-conjugated dendrimers were investigated by micro positron emission tomography and multispectral optoacoustic tomography imaging techniques and compared with that of GEM-conjugated poly(amidoamine) (PAMAM). The GEM-conjugated dendrimer with the longest peripheral PEG segments exhibited the most desirable tumor accumulation and penetration and thus had significantly higher antitumor activity than the GEM-conjugated PAMAM.


Directed induction of functional motor neuron-like cells from genetically engineered human mesenchymal stem cells.
Cell replacement using stem cells is a promising therapeutic approach to treat degenerative motor neuron (MN) disorders, such as amyotrophic lateral sclerosis and spinal cord injury. Human bone marrow-derived mesenchymal stem cells (hMSCs) are a desirable cell source for autologous cell replacement therapy to treat nervous system injury due to their plasticity, low immunogenicity, and a lower risk of tumor formation than embryonic stem cells. However, hMSCs are inefficient with regards to differentiating into MN-like cells. To solve this limitation, we genetically engineered hMSCs to express MN-associated transcription factors, Olig2 and Hb9, and then treat the hMSCs expressing Olig2 and Hb9 with optimal MN induction medium (MNIM). This method of induction led to higher expression (>30% of total cells) of MN markers. Electrophysiological data revealed that the induced hMSCs had the excitable properties of neurons and were able to form functional connections with muscle fibers in vitro. Furthermore, when the induced hMSCs were transplanted into an injured organotypic rat spinal cord slice culture, an ex vivo model of spinal cord injury, they exhibited characteristics of MNs. The data strongly suggest that induced Olig2/Hb9-expressing hMSCs were clearly reprogrammed and directed toward a MN-like lineage. We propose that methods to induce Olig2 and Hb9, followed by further induction with MNIM have therapeutic potential for autologous cell replacement therapy to treat degenerative MN disorders.


Axonal pathways to the lateral superior olive labeled with biotinylated dextran amine injections in the dorsal cochlear nucleus of rats.
The lateral superior olive (LSO) contains cells that are sensitive to intensity differences between the two ears, a feature used by the brain to localize sounds in space. This report describes a source of input to the LSO that complements bushy cell projections from the ventral cochlear nucleus (VCN). Injections of biotinylated dextran amine (BDA) into the dorsal cochlear nucleus (DCN) of the rat label axons and swellings in several brainstem structures, including the ipsilateral LSO. Labeling in the ipsilateral LSO was confined to a thin band that extended throughout the length of the structure such that it resembled an LSO isofrequency lamina. The source of this labeled pathway was not obvious, because DCN neurons do not project to the LSO, and VCN bushy cells were not filled by these injections. Filled neurons in several brainstem structures emerged as possible sources. Three observations suggest that most of the axonal labeling in the LSO derives from a single source. First, the number of labeled VCN planar multipolar cells and the amount of labeling in the LSO were consistent and robust across animals. In contrast, the number of labeled cells in most other structures was small and highly variable. Second, the locations of planar cells and filled axons in the LSO were related topographically to the position of the DCN injection site. Third, labeled terminal arborizations in the LSO arose from collaterals of axons in the trapezoid body (output tract of planar cells). We infer that planar multipolar cells, in addition to bushy cells, are a source of ascending input from the cochlear nucleus to the LSO.


Career Choice and Primary Care in the United Arab Emirates.
The low number of medical trainees entering primary care is contributing to the lack of access to primary care services in many countries. Despite the need for primary care physicians in the Middle East, there is limited information regarding trainees' career choices, a critical determinant in the supply of primary care physicians. We analyzed the career choices of medical students in the United Arab Emirates (UAE), with a larger goal of reforming postgraduate training in the region and enhancing the focus on primary care. We conducted a cross-sectional survey of applicants to a large established internal medicine residency program in the UAE. We calculated data for demographics, subspecialty choice, and factors affecting subspecialty choice, and we also reported descriptive statistics. Our response rate was 86% (183 of 212). Only 25% of applicants (n = 46) were interested in general internal medicine. The majority of respondents (n = 126, 69%) indicated a desire to pursue subspecialty training, and the remainder chose careers in research or administration. A majority of respondents (73%) were women, unmarried, and childless. Educational debt or lifestyle were not indicated as important factors in career choice. Low interest in primary care was similar to that in many Western countries, despite a much higher percentage of female applicants and a reduced emphasis on lifestyle or income factors in career decisions. Reasons for the reduced interest in primary care deserve further exploration, as do tests of interventions to increase interest, such as improving the primary care clerkship experience.


Microbicides in the prevention of HIV infection: current status and future directions.
More than 28 years since the first cases of HIV/AIDS, there is still no cure or vaccine. The worst affected region is sub-Saharan Africa and, increasingly, it is young women who are bearing the brunt of the epidemic. Consequently, there is an urgent need for HIV prevention options for women in developing countries. Microbicides are topical products that can be used vaginally by women to impede sexual transmission of HIV and thus represent one of the most promising prevention strategies. Efficacy trials with early nonspecific microbicide gels have so far been unsuccessful, but the field has now switched its focus to products containing highly potent and highly specific antiretroviral drugs that are easier to use, and can be formulated in a variety of dosage forms to suit individual and regional preferences. However, these products have their own challenges, with a greater likelihood of absorption, and the potential for systemic toxicities or the development of resistance in infected individuals who are unaware of their HIV status. The conduct of clinical trials is complex for all microbicides, with limited availability of trial sites, difficulties in dose selection and safety monitoring, and a lack of a truly objective measure of adherence. Once a microbicide has been shown to be safe and effective, there will need to be a clear pathway to regulatory approval, and the successful launch of a product will depend on having in place appropriate methods for distribution to the women who need it, along with a strategy for ensuring that they use it correctly. This will require substantial effort in terms of education and community engagement, and these activities need to be initiated well in advance of microbicide rollout.


Radiation inactivation of galactose oxidase, a monomeric enzyme with a stable free radical.
To determine the radiation sensitivity of galactose oxidase, a 68 kDa monomeric enzyme containing a mononuclear copper ion coordinated with an unusually stable cysteinyl-tyrosine (Cys-Tyr) protein free radical. Both active enzyme and reversibly rendered inactive enzyme were irradiated in the frozen state with high-energy electrons. Surviving polypeptides and surviving enzyme activity were analyzed by radiation target theory giving the radiation sensitive mass for each property. In both active and inactive forms, protein monomer integrity was lost with a single radiation interaction anywhere in the polypeptide, but enzymatic activity was more resistant, yielding target sizes considerably smaller than that of the monomer. These results suggest that the structure of galactose oxidase must make its catalytic activity unusually robust, permitting the enzymatic properties to survive in molecules following cleavage of the polymer chain. Radiation target size for loss of monomers yielded the mass of monomers indicating a polypeptide chain cleavage after a radiation interaction anywhere in the monomer. Loss of enzymatic activity yielded a much smaller mass indicating a robust structure in which catalytic activity could be expressed in cleaved polypeptides.


A comparative study of the localization of wheat-germ agglutinin and its potential receptors in wheat grains.
Wheat-germ agglutinin is located only in the embryo of a dry wheat seed and not in the endosperm tissue. This distribution remains unaltered for up to 96 h of germination and growth. The lectin is found not only in a freely soluble form but also in reversible association with particulate subcellular components. There appear to be no poly-peptides that can be solubilized with sonication and aqueous buffers from the embryo tissue that can interact with the agglutinin. This suggests that in vivo the lectin remains uncomplexed to endogenous glycoconjugates or is only able to bind to glycosylated integral membrane polypeptides. Alternatively the potential endogenous receptor(s) to wheat-germ agglutinin may not contain a polypeptide. Although the lectin is not present in the endosperm, seven polypeptides able to interact in a reversible way with wheat-germ agglutinin could be purified from that tissue.


[Tumor thrombosis in inferior vena cava: diagnostic imaging and therapeutic approximation].
Review of our experience in the diagnosis and treatment of 44 patients with inferior vena cava tumoral thrombosis (IVCTT), associated or not to other neoplastic processes: 34 hypernephroma, 2 cava leiomyosarcoma, 1 paratesticular rhabdomyosarcoma and 1 biphasic synovial sarcoma. Twenty-five patients with hypernephroma and tumor thrombi in the ipsilateral renal vein only were excluded from the analysis since this fact did not change the usual therapeutic approach. In the 19 remaining patients, concomitantly to the primary tumour exeresis a thrombectomy was performed, using cavotomy with proximal and distal clamping in 11 patients and cardiopulmonary by-pass, deep hypothermia and cardiocirculatory arrest in 8 patients. The paper analyzes the radiological investigations performed in order to reach a IVCTT diagnosis, and reviews the related literature.


Guglielmi detachable coil embolization for unruptured aneurysms in nonsurgical candidates: a cost-effectiveness exploration.
We calculated the incremental cost-utility ratio for Guglielmi detachable coil (GDC) embolization versus no therapy for unruptured intracranial aneurysms considered inappropriate for surgical clipping procedures. Decision tree and Markov analyses that employ cohort simulation were applied to determine the incremental cost-utility ratio of GDC embolization versus no therapy for unruptured cerebral aneurysms. Clinical values required as input data were estimated from the literature for the following variables: relative frequencies of complete aneurysmal occlusion, partial aneurysmal occlusion, and attempted coiling (no coils detached); morbidity and mortality of GDC embolization; frequency, morbidity, and mortality of spontaneous aneurysmal rupture in untreated and GDC-embolized aneurysms; annual rate of recanalization of GDC-embolized aneurysms; quality of life when knowingly living with untreated or GDG-embolized aneurysms and of living with fixed neurologic deficit; costs of GDC embolization, spontaneous aneurysmal rupture, stroke, and rehabilitation; and discount rate. Cost-utility ratios below $50000 per quality-adjusted life year saved were considered acceptable. Sensitivity analyses were performed for all relevant input variables. Baseline input values resulted in acceptable cost-utility ratios for GDC embolization of unruptured intracranial aneurysms. These ratios remained within acceptable limits across wide ranges of various input parameters. Cost-effectiveness was markedly affected by the natural course of unruptured, untreated aneurysms; rates of spontaneous rupture greater than 2% per year resulted in favorable cost-utility ratios that were relatively unaffected by variation in GDC efficacy, while rates of rupture less than 1% per year resulted in unfavorable ratios that were highly dependent on GDC efficacy. Many of the GDC efficacy indexes, such as rate of failed coiling, early recanalization, and progressive aneurysmal thrombosis, have mild effects on the cost-utility ratios. GDC complication rate as well as life expectancy had moderate effects on the analysis. The influence of late aneurysmal recanalization was mild unless high rates of rupture for partially coiled aneurysms were applied. Suboptimal clip placement resulting from the presence of GDC coils within a ruptured aneurysm had no demonstrable consequence on cost-utility ratios. The single most influential variable determining the cost-effectiveness of GDC embolization in our analysis was the natural course of untreated aneurysms. Other important variables included GDC-related morbidity and life expectancy at the time of GDC embolization.


Substituent Effects on Exchange Coupling and Magnetic Relaxation in 2,2'-Bipyrimidine Radical-Bridged Dilanthanide Complexes.
Systematic analysis of related compounds is crucial to the design of single-molecule magnets with improved properties, yet such studies on multinuclear lanthanide complexes with strong magnetic coupling remain rare. Herein, we present the synthesis and magnetic characterization of the series of radical-bridged dilanthanide complex salts [(Cp*2Ln)2(μ-5,5'-R2bpym)](BPh4) (Ln = Gd, Dy; R = NMe2 (1), OEt (2), Me (3), F (4); bpym = 2,2'-bipyrimidine). Modification of the substituent on the bridging 5,5'-R2bpym radical anion allows the magnetic exchange coupling constant, JGd-rad, for the gadolinium compounds in this series to be tuned over a range from -2.7 cm-1 (1) to -11.1 cm-1 (4), with electron-withdrawing or -donating substituents increasing or decreasing the strength of exchange coupling, respectively. Modulation of the exchange coupling interaction has a significant impact on the magnetic relaxation dynamics of the single-molecule magnets 1-Dy through 4-Dy, where stronger JGd-rad for the corresponding Gd3+ compounds is associated with larger thermal barriers to magnetic relaxation (Ueff), open magnetic hysteresis at higher temperatures, and slower magnetic relaxation rates for through-barrier processes. Further, we derive an empirical linear correlation between the experimental Ueff values for 1-Dy through 4-Dy and the magnitude of JGd-rad for the corresponding gadolinium derivatives that provides insight into the electronic structure of these complexes. This simple model applies to other organic radical-bridged dysprosium complexes in the literature, and it establishes clear design criteria for increasing magnetic operating temperatures in radical-bridged molecules.


Global population genetic dynamics of a highly migratory, apex predator shark.
Knowledge of genetic connectivity dynamics in the world's large-bodied, highly migratory, apex predator sharks across their global ranges is limited. One such species, the tiger shark (Galeocerdo cuvier), occurs worldwide in warm temperate and tropical waters, uses remarkably diverse habitats (nearshore to pelagic) and possesses a generalist diet that can structure marine ecosystems through top-down processes. We investigated the phylogeography and the global population structure of this exploited, phylogenetically enigmatic shark by using 10 nuclear microsatellites (n = 380) and sequences from the mitochondrial control region (CR, n = 340) and cytochrome oxidase I gene (n = 100). All three marker classes showed the genetic differentiation between tiger sharks from the western Atlantic and Indo-Pacific ocean basins (microsatellite FST > 0.129; CR ΦST > 0.497), the presence of North vs. southwestern Atlantic differentiation and the isolation of tiger sharks sampled from Hawaii from other surveyed locations. Furthermore, mitochondrial DNA revealed high levels of intraocean basin matrilineal population structure, suggesting female philopatry and sex-biased gene flow. Coalescent- and genetic distance-based estimates of divergence from CR sequences were largely congruent (dcorr = 0.0015-0.0050), indicating a separation of Indo-Pacific and western Atlantic tiger sharks <1 million years ago. Mitochondrial haplotype relationships suggested that the western South Atlantic Ocean was likely a historical connection for interocean basin linkages via the dispersal around South Africa. Together, the results reveal unexpectedly high levels of population structure in a highly migratory, behaviourally generalist, cosmopolitan ocean predator, calling for management and conservation on smaller-than-anticipated spatial scales.


Maternal condition, yolk androgens and offspring performance: a supplemental feeding experiment in the lesser black-backed gull (Larus fuscus).
It has been proposed that the maternal androgens in avian egg yolk enhance offspring fitness by accelerating growth and improving competitive ability. Because egg quality is strongly influenced by maternal condition, we predicted that females in good condition would produce high-quality eggs with relatively high androgen content. We experimentally enhanced maternal condition by supplementary feeding lesser black-backed gulls (Larus fuscus) during egg formation and compared the concentrations of androstenedione (A4), 5alpha-dihydrotestosterone (DHT) and testosterone (T) in their eggs with those in eggs laid by control females. We also measured circulating levels of T in females immediately after laying. Egg androgens could affect offspring performance directly through chick development and/or indirectly through changes in the competitive ability of a chick relative to its siblings. To avoid confounding these two routes, and to separate effects operating through the egg itself with those operating through experimental changes in parental chick rearing capacity, we fostered eggs from both maternal treatment groups singly into the nests of unmanipulated parents. Contrary to expectation, mothers with experimentally enhanced body condition laid eggs with lower levels of androgens, while exhibiting higher circulating T concentrations post-laying. Despite these lower levels of egg androgen, offspring hatched from eggs laid by mothers in good condition did not show reduced growth or survival when reared in the absence of sibling competition. Our results demonstrate that yolk androgen concentrations vary with the body condition of the female at the time of egg formation and that females in good condition reduced the yolk androgen content of their eggs without altering offspring performance.


The occurrence of Toxocara malaysiensis in cats in China, confirmed by sequence-based analyses of ribosomal DNA.
Non-isotopic polymerase chain reaction (PCR)-based single-strand conformation polymorphism and sequence analyses of the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA) were utilized to genetically characterise ascaridoids from dogs and cats from China by comparison with those from other countries. The study showed that Toxocara canis, Toxocara cati, and Toxascaris leonina from China were genetically the same as those from other geographical origins. Specimens from cats from Guangzhou, China, which were morphologically consistent with Toxocara malaysiensis, were the same genetically as those from Malaysia, with the exception of a polymorphism in the ITS-2 but no unequivocal sequence difference. This is the first report of T. malaysiensis in cats outside of Malaysia (from where it was originally described), supporting the proposal that this species has a broader geographical distribution. The molecular approach employed provides a powerful tool for elucidating the biology, epidemiology, and zoonotic significance of T. malaysiensis.


Effect of chelation therapy on the neuropsychological and behavioral development of lead-exposed children after school entry.
Some children in the United States continue to be exposed to levels of lead that increase their risk for lowered intellectual functioning and behavior problems. It is unclear whether chelation therapy can prevent or reverse the neurodevelopmental sequelae of lead toxicity. The objective of this study was to determine whether chelation therapy with succimer (dimercaptosuccinic acid) in children with referral blood lead levels between 20 and 44 microg/dL (0.96-2.12 micromol/L) at 12 to 33 months of age has neurodevelopmental benefits at age 7 years. The Treatment of Lead-Exposed Children (TLC) study is a randomized, double-blind, placebo-controlled trial that was conducted between September 1994 and June 2003 in Philadelphia, PA; Newark, NJ; Cincinnati, OH; and Baltimore, MD. Of 1854 referred children who were between the ages of 12 to 33 months and screened for eligibility, 780 were randomized to the active drug and placebo groups stratified by clinical center, body surface area, blood lead level, and language spoken at home. At 7 years of age, 647 subjects remained in the study. Participants were randomly assigned to receive oral succimer or placebo. Up to 3 26-day courses of succimer or placebo therapy were administered depending on response to treatment in those who were given active drug. Eighty-nine percent had finished treatment by 6 months, with all children finishing by 13 months after randomization. All participants received residential lead hazard control measures before treatment. TLC subjects also received a daily multivitamin supplement before and after treatment(s) with succimer or placebo. Scores on standardized neuropsychological measures that tap cognition, behavior, learning and memory, attention, and neuromotor skills were measured. Chelation therapy with succimer lowered average blood lead levels for approximately 6 months but resulted in no benefit in cognitive, behavioral, and neuromotor endpoints. These new follow-up data confirm our previous finding that the TLC regimen of chelation therapy is not associated with neurodevelopmental benefits in children with blood lead levels between 20 and 44 microg/dL (0.96-2.17 micromol/L). These results emphasize the importance of taking environmental measures to prevent exposure to lead. Chelation therapy with succimer cannot be recommended for children with blood lead levels between 20 and 44 microg/dL (0.96-2.12 micromol/L).


Cell cycle dependence of cytotoxicity and clastogenicity induced by treatment of synchronized human diploid fibroblasts with sodium fluoride.
To study the cell cycle dependence of cytotoxicity and clastogenicity of sodium fluoride (NaF), synchronized human diploid fibroblasts were treated with NaF during different phases of the cell cycle and analyzed. Exponentially growing cells were synchronized by the following two procedures. (1) The cells were synchronized at G0/G1 phase by a period of growth in medium containing 1% serum (low serum medium). (2) The cells were synchronized at the G1/S boundary by growth in low serum medium, followed by hydroxyurea treatment (Tsutsui et al., 1984a). Synchronized cells were treated with NaF for 3 h during the G1 phase or G2 phase, and for each of three 3-h periods during the S phase which lasted 9 h. Cytotoxicity, as determined by a decrease in colony-forming ability, was dependent upon the phase of the cell cycle during which NaF treatment was administered. The highest lethality was induced in cultures treated with NaF during the first 3 h of S phase (early S phase). Moderate lethality was observed when the cultures were treated with NaF during the second or third 3 h of S phase (middle or late S phase, respectively), or G2 phase. Little lethality was observed in cultures in G1 phase. Inducibility of chromosome aberrations of the cells following treatment with NaF was also dependent upon the phase of the cell cycle. A significant increase in the incidence of chromosome aberrations was observed only in cultures treated with NaF during early and/or middle S phases of cell cycle. These results suggest that cytotoxicity and clastogenicity of NaF to cultured human diploid fibroblasts are cell cycle dependent, and that the cells in early and middle S phases are more sensitive to the effects.


From pure compounds to complex exposure: Effects of dietary cadmium and lignans on estrogen, epidermal growth factor receptor, and mitogen activated protein kinase signaling in vivo.
Exposure to environmental endocrine active compounds correlates with altered susceptibility to disease in human populations. Chemical risk assessment is single compound based, although exposure often takes place as heterogeneous mixtures of man-made and natural substances within complex matrices like diet. Here we studied whether the effects of cadmium and enterolactone on endocrine endpoints in dietary exposure can be predicted based on pure compound effects. Ovariectomized estrogen reporter ERE-luciferase (ERE-luc) mice were maintained on diets that intrinsically contain increasing concentrations of cadmium and enterolactone precursors for three and 21 days. The activation of the ERE-luc, epidermal growth factor receptor (EGFR), mitogen activated protein kinase (MAPK)-ERK1/2, and classical estrogen responses were measured. Interactions between the diets and endogenous hormone were evaluated by challenging the animals with 17β-estradiol. Compared to animals on basal purified diet, mice consuming experimental diets were exposed to significantly higher levels of cadmium and enterolactone, yet the exposure remained comparable to typical human dietary intake. Surprisingly, we could not detect effects on endpoints regulated by pure enterolactone, such as ERE-luc activation. However, cadmium accumulation in the liver was accompanied with activation of EGFR and MAPK-ERK1/2 in line with our earlier CdCl2 studies. Further, attenuation of 17β-estradiol-induced ERE-luc response in liver by experimental diets was observed. Our findings indicate that the exposure context can have substantial effects on the activity of endocrine active compounds in vivo. Thus, whenever possible, a context that mimics human exposure should be tested along with pure compounds.


Preferential lentiviral targeting of astrocytes in the central nervous system.
The ability to visualize and genetically manipulate specific cell populations of the central nervous system (CNS) is fundamental to a better understanding of brain functions at the cellular and molecular levels. Tools to selectively target cells of the CNS include molecular genetics, imaging, and use of transgenic animals. However, these approaches are technically challenging, time consuming, and difficult to control. Viral-mediated targeting of cells in the CNS can be highly beneficial for studying and treating neurodegenerative diseases. Yet, despite specific marking of numerous cell types in the CNS, in vivo selective targeting of astrocytes has not been optimized. In this study, preferential targeting of astrocytes in the CNS was demonstrated using engineered lentiviruses that were pseudotyped with a modified Sindbis envelope and displayed anti-GLAST IgG on their surfaces as an attachment moiety. Viral tropism for astrocytes was initially verified in vitro in primary mixed glia cultures. When injected into the brains of mice, lentiviruses that displayed GLAST IgG on their surface, exhibited preferential astrocyte targeting, compared to pseudotyped lentiviruses that did not incorporate any IgG or that expressed a control isotype IgG. Overall, this approach is highly flexible and can be exploited to selectively target astrocytes or other cell types of the CNS. As such, it can open a window to visualize and genetically manipulate astrocytes or other cells of the CNS as means of research and treatment.


Comparative analysis of methods for detecting interacting loci.
Interactions among genetic loci are believed to play an important role in disease risk. While many methods have been proposed for detecting such interactions, their relative performance remains largely unclear, mainly because different data sources, detection performance criteria, and experimental protocols were used in the papers introducing these methods and in subsequent studies. Moreover, there have been very few studies strictly focused on comparison of existing methods. Given the importance of detecting gene-gene and gene-environment interactions, a rigorous, comprehensive comparison of performance and limitations of available interaction detection methods is warranted. We report a comparison of eight representative methods, of which seven were specifically designed to detect interactions among single nucleotide polymorphisms (SNPs), with the last a popular main-effect testing method used as a baseline for performance evaluation. The selected methods, multifactor dimensionality reduction (MDR), full interaction model (FIM), information gain (IG), Bayesian epistasis association mapping (BEAM), SNP harvester (SH), maximum entropy conditional probability modeling (MECPM), logistic regression with an interaction term (LRIT), and logistic regression (LR) were compared on a large number of simulated data sets, each, consistent with complex disease models, embedding multiple sets of interacting SNPs, under different interaction models. The assessment criteria included several relevant detection power measures, family-wise type I error rate, and computational complexity. There are several important results from this study. First, while some SNPs in interactions with strong effects are successfully detected, most of the methods miss many interacting SNPs at an acceptable rate of false positives. In this study, the best-performing method was MECPM. Second, the statistical significance assessment criteria, used by some of the methods to control the type I error rate, are quite conservative, thereby limiting their power and making it difficult to fairly compare them. Third, as expected, power varies for different models and as a function of penetrance, minor allele frequency, linkage disequilibrium and marginal effects. Fourth, the analytical relationships between power and these factors are derived, aiding in the interpretation of the study results. Fifth, for these methods the magnitude of the main effect influences the power of the tests. Sixth, most methods can detect some ground-truth SNPs but have modest power to detect the whole set of interacting SNPs. This comparison study provides new insights into the strengths and limitations of current methods for detecting interacting loci. This study, along with freely available simulation tools we provide, should help support development of improved methods. The simulation tools are available at: http://code.google.com/p/simulation-tool-bmc-ms9169818735220977/downloads/list.


Heparin-induced thrombocytopenia presenting as splenic hemorrhage following cardiac surgery: a case report.
Heparin-induced thrombocytopenia with thrombosis (HITT) is a paradoxical prothrombotic complication of anticoagulant therapy. As many as 3% of patients undergoing cardiac surgery develop clinical HIT presenting as thrombocytopenia with or without thrombosis within 5-10 days of heparin exposure. Thrombotic complications associated with HIT carry a mortality rate of 5-10%. We report a case of atraumatic splenic hemorrhage due to splenic vein thrombosis as the main indicator of HIT following cardiac surgery in a 62-year-old woman. She presented to the emergency department on day nine following coronary artery bypass graft surgery with acute weakness, dizziness, and malaise. Her evaluation in the emergency department found anemia and thrombocytopenia. A coagulation profile revealed a markedly elevated d-dimer. She underwent a computed tomography scan of the chest, abdomen and pelvis for suspected bleed and was found to have splenic vein thrombosis, right atrial filling defects consistent with atrial thrombus and mild to moderate hemoperitoneum. Surgical consultation was obtained due to splenic hemorrhage. Hematology was consulted on post-operative day 10, however, she unfortunately developed left sided weakness concerning for stroke. A magnetic resonance imaging scan of the brain demonstrated infarct involving distribution of the right anterior cerebral artery. A transesophageal echocardiogram demonstrated a large immobile thrombus within the right atrium with a second, mobile thrombus arising from the left tricuspid valve annulus. Due to a 4Ts score of 7 and markedly positive platelet factor 4 (PF4) IgG antibody a serotonin release assay was not performed given the high probability of HIT. She was cautiously treated with bivalirudin and was transitioned to warfarin anticoagulation. In the following days her platelet count recovered and 3 months later a transthoracic echocardiogram revealed solution of the intracardiac thrombi. Atraumatic splenic hemorrhage is an unusual presentation of HIT that is reminiscent of the rare bilateral adrenal hemorrhage due to adrenal necrosis that also occurs in HIT. Alternative anticoagulation is the mainstay of therapy for HIT despite hemorrhage, given the underlying acquired hypercoagulability. Despite similarities of the presentation between splenic hemorrhage and bilateral adrenal hemorrhage, splenic hemorrhage is rarely described in the literature. HIT should be considered in patients presenting with thrombocytopenia following cardiac surgery.


Emergence of oscillations in a simple epidemic model with demographic data.
A simple susceptible-infectious-removed epidemic model for smallpox, with birth and death rates based on historical data, produces oscillatory dynamics with remarkably accurate periodicity. Stochastic population data cause oscillations to be sustained rather than damped, and data analysis regarding the oscillations provides insights into the same set of population data. Notably, oscillations arise naturally from the model, instead of from a periodic forcing term or other exogenous mechanism that guarantees oscillation: the model has no such mechanism. These emergent natural oscillations display appropriate periodicity for smallpox, even when the model is applied to different locations and populations. The model and datasets, in turn, offer new observations about disease dynamics and solution trajectories. These results call for renewed attention to relatively simple models, in combination with datasets from real outbreaks.


The school setting: an opportunity for the implementation of dietary guidelines.
Nutrition, physical activity and health related promotion programmes in schools have developed into a mature field of research over the past decades. A number of success factors have been identified and evidence-based interventions have been performed. However, the school setting as an arena for evidence-based health promotion programmes, is still not used to its full potential. Schools provide an excellent arena for reaching large segments of the population, such as young people, school staff, families and the surrounding community. There is a need for an overview regarding the current status of nutrition, physical activity, related health as well as support structures in the EU member states. Based upon such an analysis, a consensus report should be written, pointing out the major problems at hand. Self-assessment tools for national as well as for school level should be produced, in order to guide changes, but also to include an element of continuous assessment of change, for evaluation purposes.


North Carolina Medicaid recipient management lock-in program: the pharmacist's perspective.
The misuse and abuse of prescription opioids have become an urgent health issue in North Carolina (NC), particularly among Medicaid patients who suffer high rates of morbidity and mortality due to abuse and overdose. The NC Division of Medical Assistance (DMA) implemented a recipient management lock-in program, which limits identified patients for a 12-month period to 1 prescriber and 1 pharmacy for benzodiazepine, opiate, and certain anxiolytic prescriptions in order to prevent misuse and reduce overutilization of Medicaid benefits. To (a) evaluate pharmacists' perceptions of the implementation of the NC recipient management lock-in program (MLIP) and (b) determine how the beliefs and attitudes of pharmacists could promote or inhibit its success. We conducted 12 structured phone interviews with NC pharmacists serving lock-in patients. Interview responses were analyzed through construct analysis, which identified themes organized into 3 domains: organization and implementation, perceived effectiveness, and acceptability. Most respondents reported a positive experience with the program but expressed doubt concerning its impact on prescription drug abuse. The program successfully utilized the pharmacist role as a gatekeeper of controlled substances, and the procedures of the program required no active effort on pharmacists' part. However, respondents suggested that the DMA improve communication and outreach to address pharmacists' lack of knowledge about the program's purpose and confusion over remediating problems that arise with lock-in patients. The DMA should also address the ways in which the program can interfere with access to health care and treatment, allow patients to see multiple physicians within the same clinic, and clarify procedures for patients whose complex health issues require multiple specialists. Although possible improvements were identified, the NC MLIP has strong potential for success as it utilizes pharmacists' medication gate-keeping role, while minimizing the effort required for successful implementation.


Assessing and understanding sedentary behaviour in office-based working adults: a mixed-method approach.
Sedentary behaviours (SB) can be characterized by low energy expenditure in a reclining position (e.g., sitting) often associated with work and transport. Prolonged SB is associated with increased risk for chronic conditions, and due to technological advances, the working population is in office settings with high occupational exposure to SB. This study aims to assess SB among office workers, as well as barriers and strategies towards reducing SB in the work setting. Using a mixed-methods approach guided by the socio-ecological framework, non-academic office workers from a professional school in a large public university were recruited. Of 180 eligible office workers, 40 enrolled and completed all assessments. Self-reported and objectively measured SB and activity levels were captured. Focus group discussion (FGD) were conducted to further understand perceptions, barriers, and strategies to reducing workplace SB. Environmental factors were systematically evaluated by trained research staff using an adapted version of the Checklist for Health Promotion Environments at Worksites (CHEW). Thematic analysis of FGD was conducted and descriptive analysis of quantitative data was performed. The sample was mostly Chinese (n = 33, 80 %) with a total of 24 (60 %) female participants. Most participants worked five days a week for about 9.5(0.5) hrs/day. Accelerometer data show that participants spend the majority of their days in sedentary activities both on workdays (76.9 %) and non-workdays (69.5 %). Self-report data confirm these findings with median sitting time of 420(180) minutes at work. From qualitative analyses, major barriers to reducing SB emerged, including the following themes: workplace social and cultural norms, personal factors, job scope, and physical building/office infrastructure. CHEW results confirm a lack of support from the physical infrastructure and information environment to reducing SB. There is high SB among office workers in this sample. We identified multiple levels of influence for prolonged occupational SB, with a particular emphasis on workplace norms and infrastructure as important barriers to reducing SB and increasing PA. A larger, representative sample of the Singaporean population is needed to confirm our findings but it seems that any intervention aimed at reducing SB in the workplace should target individual, environmental, and organizational levels.


Intraoperative evaluation of device placement in spine surgery using known-component 3D-2D image registration.
Intraoperative x-ray radiography/fluoroscopy is commonly used to assess the placement of surgical devices in the operating room (e.g. spine pedicle screws), but qualitative interpretation can fail to reliably detect suboptimal delivery and/or breach of adjacent critical structures. We present a 3D-2D image registration method wherein intraoperative radiographs are leveraged in combination with prior knowledge of the patient and surgical components for quantitative assessment of device placement and more rigorous quality assurance (QA) of the surgical product. The algorithm is based on known-component registration (KC-Reg) in which patient-specific preoperative CT and parametric component models are used. The registration performs optimization of gradient similarity, removes the need for offline geometric calibration of the C-arm, and simultaneously solves for multiple component bodies, thereby allowing QA in a single step (e.g. spinal construct with 4-20 screws). Performance was tested in a spine phantom, and first clinical results are reported for QA of transpedicle screws delivered in a patient undergoing thoracolumbar spine surgery. Simultaneous registration of ten pedicle screws (five contralateral pairs) demonstrated mean target registration error (TRE) of 1.1 ± 0.1 mm at the screw tip and 0.7 ± 0.4° in angulation when a prior geometric calibration was used. The calibration-free formulation, with the aid of component collision constraints, achieved TRE of 1.4 ± 0.6 mm. In all cases, a statistically significant improvement (p < 0.05) was observed for the simultaneous solutions in comparison to previously reported sequential solution of individual components. Initial application in clinical data in spine surgery demonstrated TRE of 2.7 ± 2.6 mm and 1.5 ± 0.8°. The KC-Reg algorithm offers an independent check and quantitative QA of the surgical product using radiographic/fluoroscopic views acquired within standard OR workflow. Such intraoperative assessment could improve quality and safety, provide the opportunity to revise suboptimal constructs in the OR, and reduce the frequency of revision surgery.


The Luoping biota: exceptional preservation, and new evidence on the Triassic recovery from end-Permian mass extinction.
The timing and nature of biotic recovery from the devastating end-Permian mass extinction (252 Ma) are much debated. New studies in South China suggest that complex marine ecosystems did not become re-established until the middle-late Anisian (Middle Triassic), much later than had been proposed by some. The recently discovered exceptionally preserved Luoping biota from the Anisian Stage of the Middle Triassic, Yunnan Province and southwest China shows this final stage of community assembly on the continental shelf. The fossil assemblage is a mixture of marine animals, including abundant lightly sclerotized arthropods, associated with fishes, marine reptiles, bivalves, gastropods, belemnoids, ammonoids, echinoderms, brachiopods, conodonts and foraminifers, as well as plants and rare arthropods from nearby land. In some ways, the Luoping biota rebuilt the framework of the pre-extinction latest Permian marine ecosystem, but it differed too in profound ways. New trophic levels were introduced, most notably among top predators in the form of the diverse marine reptiles that had no evident analogues in the Late Permian. The Luoping biota is one of the most diverse Triassic marine fossil Lagerstätten in the world, providing a new and early window on recovery and radiation of Triassic marine ecosystems some 10 Myr after the end-Permian mass extinction.


Does Implant Selection Affect the Inpatient Cost of Care for Geriatric Intertrochanteric Femur Fractures?
Geriatric intertrochanteric (IT) femur fractures are a common and costly injury, expected to increase in incidence as the population ages. Understanding cost drivers will be essential for risk adjustments, and the surgeon's choice of implant may be an opportunity to reduce the overall cost of care. This study was purposed to identify the relationship between implant type and inpatient cost of care for isolated geriatric IT fractures. A retrospective review of IT fractures from 2013-2017 was performed at an academic level I trauma center. Construct type and AO/OTA fracture classifications were obtained radiographically, and patient variables were collected via the electronic medical record (EMR). The total cost of care was obtained via time-driven activity-based costing (TDABC). Multivariable linear regression and goodness-of-fit analyses were used to determine correlation between implant costs, inpatient cost of care, construct type, patient characteristics, and injury characteristics. Implant costs ranged from $765.17 to $5,045.62, averaging $2,699, and were highest among OTA 31-A3 fracture patterns (p < 0.01). Implant cost had a positive linear association with overall inpatient cost of care (p < 0.01), but remained highly variable (r2 = 0.16). Total cost of care ranged from $9,129.18 to $64,210.70, averaging $19,822, and patients receiving a sliding hip screw (SHS) had the lowest mean total cost of care at $17,077, followed by short and long intramedullary nails ($19,314 and $21,372, respectively). When construct type and fracture pattern were compared to total cost, 31-A1 fracture pattern treated with SHS had significantly lower cost than 31-A2 and 31-A3 and less variation in cost. The cost of care for IT fractures is poorly understood and difficult to determine. With alternative payment models on the horizon, implant selection should be utilized as an opportunity to decrease costs and increase the value of care provided to patients. Diagnostic Level IV.


The role of Government policy in supporting nurse-led care in general practice in the United Kingdom, New Zealand and Australia: an adapted realist review.
This article is a report on a review that examined the role of Government policy in primary care and its association with nurse-led care in the United Kingdom, New Zealand and Australia between 1998 and 2009. The United Kingdom, New Zealand and Australia share a similar model of first point access to the healthcare system via general practitioners. General practice is synonymous with the term primary care. DATA OURCES: Medline, CINAHL, EMBASE, Scopus, PsychInfo, Google, Department of Health, England (United Kingdom), Ministry of Health, New Zealand, Department of Health and Ageing, Australia. Searches of electronic databases from 1998 to December 2009 and hand searches of identified leads and key journals. Historical papers accessed to describe the genesis of practice nursing and historical Government policy documents prior to 1998, were examined. A modified realist review was used to synthesize research and policy documents relating to government policies pertaining to nurse-led care. In addition, a systematic review was used to identify literature that described practice nurse-led care. Results. Nurse-led primary care services are well described in the United Kingdom with a total of 45 studies meeting the inclusion criteria for the second review. There are no published studies from New Zealand, and only two from Australia describing nurse-led primary care. New Zealand and Australia lag behind the United Kingdom in practice nurse development. Implementation of clinical governance was fundamental to the development of nurse-led care in the UK.


Intracellular reactive oxygen species mediate the linkage of Na+/K+-ATPase to hypertrophy and its marker genes in cardiac myocytes.
We showed before that in cardiac myocytes partial inhibition of Na+/K+-ATPase by nontoxic concentrations of ouabain causes hypertrophy and transcriptional regulations of growth-related marker genes through multiple Ca2+-dependent signal pathways many of which involve Ras and p42/44 mitogen-activated protein kinases. The aim of this work was to explore the roles of intracellular reactive oxygen species (ROS) in these ouabain-initiated pathways. Ouabain caused a rapid generation of ROS within the myocytes that was prevented by preexposure of cells to N-acetylcysteine (NAC) or vitamin E. These antioxidants also blocked or attenuated the following actions of ouabain: inductions of the genes of skeletal alpha-actin and atrial natriuretic factor, repression of the gene of the alpha3-subunit of Na+/K+-ATPase, activation of mitogen-activated protein kinases, activation of Ras-dependent protein synthesis, and activation of transcription factor NF-kappaB. Induction of c-fos and activation of AP-1 by ouabain were not sensitive to NAC. Ouabain-induced inhibition of active Rb+ uptake through Na+/K+-ATPase and the resulting rise in intracellular Ca2+ were also not prevented by NAC. A phorbol ester that also causes myocyte hypertrophy did not increase ROS generation, and its effects on marker genes and protein synthesis were not affected by NAC. We conclude the following: (a) ROS are essential second messengers within some but not all signal pathways that are activated by the effect of ouabain on Na+/K+-ATPase; (b) the ROS-dependent pathways are involved in ouabain-induced hypertrophy; (c) increased ROS generation is not a common response of the myocyte to all hypertrophic stimuli; and (d) it may be possible to dissociate the positive inotropic effect of ouabain from its growth-related effects by alteration of the redox state of the cardiac myocyte.


Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis.
Plant defense against microbial pathogens depends on the action of several endogenously produced hormones, including jasmonic acid (JA) and ethylene (ET). In defense against necrotrophic pathogens, the JA and ET signaling pathways synergize to activate a specific set of defense genes including PLANT DEFENSIN1.2 (PDF1.2). The APETALA2/Ethylene Response Factor (AP2/ERF)-domain transcription factor ORA59 acts as the integrator of the JA and ET signaling pathways and is the key regulator of JA- and ET-responsive PDF1.2 expression. The present study was aimed at the identification of elements in the PDF1.2 promoter conferring the synergistic response to JA/ET and interacting with ORA59. We show that the PDF1.2 promoter was activated synergistically by JA and the ET-releasing agent ethephon due to the activity of two GCC boxes. ORA59 bound in vitro to these GCC boxes and trans-activated the PDF1.2 promoter in transient assays via these two boxes. Using the chromatin immunoprecipitation technique we were able to show that ORA59 bound the PDF1.2 promoter in vivo. Finally, we show that a tetramer of a single GCC box conferred JA/ethephon-responsive expression, demonstrating that the JA and ET signaling pathways converge to a single type of GCC box. Therefore ORA59 and two functionally equivalent GCC box binding sites form the module that enables the PDF1.2 gene to respond synergistically to simultaneous activation of the JA and ET signaling pathways.


Attitudes toward genetic research review: results from a national survey of professionals involved in human subjects protection.
The recent expansion of human genetics research has raised complex ethical and regulatory issues. However, few published reports describe the views of professionals involved in human subjects protection (HSP) regarding the risks and benefits of genetic research. This anonymous, web-based study elicited the opinions of 208 HSP professionals about review of genetic research. The majority of respondents felt that different guidance is needed for various aspects of genetic protocol review compared with other types of human subjects research. Importantly, opinions were divided on specific genetic research issues, such as what constitutes human subjects research, when to re-consent, and the likelihood and risks of research participant identification. Findings from this study illustrate the need for a collaborative approach to ethics oversight in the review and conduct of genetic research.


Endothelial function in patients with obstructive sleep apnea syndrome but without hypertension.
Obstructive sleep apnea syndrome (OSAS) influences endothelial function and causes hypertension. Our aim was to evaluate the role of endothelial dysfunction in the pathogenesis of hypertension in OSAS. Twenty-three patients with OSAS but without hypertension and 15 healthy normotensive subjects were investigated. The presence or absence of OSAS was evaluated with a sleep study. Endothelial function was investigated with brachial artery ultrasound examination. Baseline characteristics were equivalent between the two groups. Minimal oxygen saturation and apnea-hypopnea indexes in the OSAS and control groups were 62.9 +/- 16.5 versus 94.9 +/- 1.1% (p < 0.0001) and 53.1 +/- 20.3 versus 3.8 +/- 0.9 (p < 0.0001), respectively. There was not statistically significant difference between basal brachial artery diameters measured in the morning and in the evening in all groups. Flow-mediated dilation (FMD) values measured in the morning were lower than those measured in the evening in both OSAS patients and the control group: FMD of OSAS patients was 6.04 +/- 3.18% in the morning and 10.38 +/- 4.23% in the evening hours (p = 0.001), and FMD of control subjects was 10.9 +/- 2.6% in the morning and 13.9 +/- 2.32 in the evening hours (p = 0.002). Differences in FMD values measured both in the morning and evening hours in OSAS patients were lower compared with those in control subjects (p < 0.0001 in the morning hours and p = 0.003 in the evening hours). We detected a prominent diurnal deterioration in endothelial function in normotensive OSAS patients compared with healthy subjects. This deterioration may occur due to ongoing hypoxemia during the night and it may be a possible cause of hypertension and atherosclerotic cardiovascular diseases in patients with OSAS.


A gateway-compatible yeast one-hybrid system.
Since the advent of microarrays, vast amounts of gene expression data have been generated. However, these microarray data fail to reveal the transcription regulatory mechanisms that underlie differential gene expression, because the identity of the responsible transcription factors (TFs) often cannot be directly inferred from such data sets. Regulatory TFs activate or repress transcription of their target genes by binding to cis-regulatory elements that are frequently located in a gene's promoter. To understand the mechanisms underlying differential gene expression, it is necessary to identify physical interactions between regulatory TFs and their target genes. We developed a Gateway-compatible yeast one-hybrid (Y1H) system that enables the rapid, large-scale identification of protein-DNA interactions using both small (i.e., DNA elements of interest) and large (i.e., gene promoters) DNA fragments. We used four well-characterized Caenorhabditis elegans promoters as DNA baits to test the functionality of this Y1H system. We could detect approximately 40% of previously reported TF-promoter interactions. By performing screens using two complementary libraries, we found novel potentially interacting TFs for each promoter. We recapitulated several of the Y1H-based protein-DNA interactions using luciferase reporter assays in mammalian cells. Taken together, the Gateway-compatible Y1H system will allow the high-throughput identification of protein-DNA interactions and may be a valuable tool to decipher transcription regulatory networks.


Stability of Cerebral Aneurysms After Stent-Assisted Coil Embolization: A Propensity Score-Matched Analysis.
The availability of stents has widened the indications of endovascular intervention for cerebral aneurysms. To elucidate the effect of stents on radiologic outcomes and to analyze the risk factors for aneurysmal recanalization via propensity score matching. From the 735 aneurysms treated with coil embolization with stents (n = 187) and without stents (n = 548) between 2009 and 2011, 157 propensity score-matched case pairs were selected. The recanalization rates and relevant risk factors were analyzed. The mean follow-up interval was 24.1 ± 11.3 months (range, 6-48 months) and 22.9 ± 11.4 months (range, 6-56 months) in the stent and nonstent groups, respectively (P = .388). The stent group demonstrated lower recanalization rates than the nonstent group during both the 6-month (1.9% vs 10.2%, P = .004) and the final follow-up periods (8.3% vs 18.5%, P = .005). The multivariate analysis identified the following significant factors for recanalization: the use of stents (hazard ratio, 0.40; 95% confidence interval, 0.21-0.76, P = .005), larger aneurysm size (hazard ratio, 1.21; 95% confidence interval, 1.11-1.31, P < .001), and initially incomplete occlusion (hazard ratio, 2.39; 95% confidence interval, 1.28-4.43, P = .006). The incidence of permanent neurological complication tended to be higher in the stent group than in the nonstent group (3.2% vs 0%, P = .063). In this propensity score-matched analysis, stent implantation reduced the overall recanalization of the coiled cerebral aneurysms. However, the use of stents should be carefully decided upon.


The Role of Ethnicity and Environment in the Regulation of Response to Sensory Stimulus in Children: Protocol and Pilot Findings of a Neurophysiological Study.
The ability to regulate the response to sensory stimuli has been associated with successful behavioral patterns necessary for daily activities. However, it is not known whether a child's ethnicity and environment can influence autonomic regulatory mechanisms. This study aims to explore the role of ethnicity and environment in the regulation of responses to sensory stimuli in children. In this study, we intend to recruit 128 children from different ethnic groups or environment contexts as follows: (1) 32 typically developing Chinese children living in Hong Kong; (2) 32 typically developing Filipino children living in Hong Kong; (3) 32 typically developing Filipino children who are living in urban areas; and (4) 32 typically developing Filipino children who are living in rural areas in Philippines. Autonomic activity (heart rate variability [HRV] and electrodermal activity [EDA]) will be measured and recorded using Polar H2 heart rate monitor and eSense GSR skin response sensor. Autonomic activity (HRV-low frequency, HRV-high frequency, and EDA) at different conditions between pairwise groupings will be tested using multivariate analysis of variance (MANOVA). All significant levels will be set at P ≤.05. We present the research protocol of this study, as well as a short discussion of the preliminary findings from our pilot data, with consequent power and sample size analysis that informs the appropriate sample needed to test our hypothesis. This study will increase the understanding on the role of individual differences related to a child's ethnicity and environment in the regulation of response to sensory stimuli. The findings of this research may further shed light on the evaluation and treatment planning for children across and within cultures.


Subcutaneous and mediastinal emphysema. Pathophysiology, diagnosis, and management.
Subcutaneous emphysema and pneumomediastinum occur frequently in critically ill patients in association with blunt or penetrating trauma, soft-tissue infections, or any condition that creates a gradient between intra-alveolar and perivascular interstitial pressures. A continuum of fascial planes connects cervical soft tissues with the medlastinum and retroperitoneum, permitting aberrant air arising in any one of these areas to spread elsewhere. Diagnosis is made in the appropriate clinical setting by careful physical examination and inspection of the chest roentgenogram. While the presence of air in subcutaneous or mediastinal tissue is not dangerous in itself, prompt recognition of the underlying cause is essential. Certain trauma-related causes may require surgical intervention, but the routine use of chest tubes tracheostomy, or mediastinal drains is not recommended.


Risk factors for drinking among HIV-positive African American adults: the depression-gender interaction.
Alcohol misuse disproportionately affects people living with HIV. People who struggle with alcohol are also likely to be concurrently struggling with depression. Although there is evidence linking depression to HIV, depression to alcohol, and alcohol to HIV, there is limited research that simultaneously examines the interlocking associations between all three factors. To investigate the interaction between depression symptomatology and gender on alcohol misuse, while controlling for motivation to reduce drinking, among HIV-positive African Americans. This population was examined because of their increased for developing alcohol dependence. Data analysis was conducted on baseline survey data of HIV-positive African American adults (N = 88) who drink and were enrolled in the Project PLUS (Positive Living through Understanding and Support) intervention to examine the correlates (i.e., gender, motivation, depression) of drinking in the past 30 days. Hierarchical linear regression analysis showed a significant interaction between gender and depression to predict total drinks reported (R(2) = .56, p < .001). While depression was the sole predictor of drinking for men and suppressed the role of motivation, the reverse was true for women; depression was not a significant predictor after controlling for motivation to reduce drinking. African American men and women living with HIV have different risk factors for recent drinking. Understanding the link between depression, gender, and motivation to reduce drinking for HIV-positive adults with alcohol problems is crucial for the development of gender and culturally relevant treatments.


The clock gene cryptochrome of Bactrocera cucurbitae (Diptera: Tephritidae) in strains with different mating times.
Differences in mating time between populations can give rise to premating reproductive isolation. Tephritid fruit flies exhibit large variation in mating time among intra- or inter-specific populations. We previously cloned the clock gene period from two strains of melon fly, Bactrocera cucurbitae; in one the individuals mate early during the day, whereas in the other the individuals mate later. These strains were originally established by divergent artificial selection for developmental time, 'short' and 'long', with early and late mating times, respectively. The deduced amino acid sequences of PERIOD proteins for these two strains were reported to be identical. Here we cloned another clock gene cryptochrome (cry) from the two strains, and found two stable amino acid substitutions in the strains. In addition, the allele frequency at the two polymorphic sites of cry gene correlated with the circadian locomotor period (tau) across strains, whereas the expression pattern of cry mRNA in the heads of flies taken from the short strain significantly differed from that from the long strain. These findings suggest that variation in the cry gene is related to differences in the circadian behaviour in the two strains, thus implying that the cry gene may have an important role in reproductive isolation.


Pronounced somatic bottleneck in mitochondrial DNA of human hair.
Heteroplasmy is the presence of variable mitochondrial DNA (mtDNA) within the same individual. The dynamics of heteroplasmy allele frequency among tissues of the human body is not well understood. Here, we measured allele frequency at heteroplasmic sites in two to eight hairs from each of 11 humans using next-generation sequencing. We observed a high variance in heteroplasmic allele frequency among separate hairs from the same individual-much higher than that for blood and cheek tissues. Our population genetic modelling estimated the somatic bottleneck during embryonic follicle development of separate hairs to be only 11.06 (95% confidence interval 0.6-34.0) mtDNA segregating units. This bottleneck is much more drastic than somatic bottlenecks for blood and cheek tissues (136 and 458 units, respectively), as well as more drastic than, or comparable to, the germline bottleneck (equal to 25-32 or 7-10 units, depending on the study). We demonstrated that hair undergoes additional genetic drift before and after the divergence of mtDNA lineages of individual hair follicles. Additionally, we showed a positive correlation between donor's age and variance in heteroplasmy allele frequency in hair. These findings have important implications for forensics and for our understanding of mtDNA dynamics in the human body. This article is part of the theme issue 'Linking the mitochondrial genotype to phenotype: a complex endeavour'.


Impact of maternal nativity on the prevalence of diabetes during pregnancy among U.S. ethnic groups.
This study examines the impact of maternal nativity (birthplace) on the overall prevalence of diabetes during pregnancy and among 15 racial and ethnic groups in the U.S. Birth certificate data for all resident single live births in the U.S. from 1994 to 1996 were used to calculate reported diabetes prevalence during pregnancy and to assess the impact of maternal birthplace outside of the 50 states and Washington, DC, on the risk of diabetes before and after adjustment for differences in maternal age, other sociodemographic characteristics, and late or no initiation of prenatal care overall and for each racial and ethnic group. Mothers born outside of the U.S. are significantly more likely to have diabetes during pregnancy. The impact of maternal nativity on diabetes prevalence is largely explained by the older childbearing age of immigrant mothers. However, adjusted diabetes risk remains elevated for Asian-Indian, non-Hispanic black, Filipino, Puerto Rican, and Central and South American mothers who were born outside the U.S. Conversely, birthplace outside the U.S. significantly reduces diabetes risk for Japanese, Mexican, and Native American women. Identification, treatment, and follow-up of immigrant mothers with diabetes during pregnancy may require special attention to language and sociocultural barriers to effective care. Systematic surveillance of the prevalence and impact of diabetes during pregnancy for immigrant and nonimmigrant women, particularly in racial and ethnic minority groups, and more detailed studies on the impact of acculturation on diabetes may increase understanding of the epidemiology of diabetes during pregnancy in our increasingly diverse society.


Milk sialyllactose influences colitis in mice through selective intestinal bacterial colonization.
Milk oligosaccharides contribute to the development of the intestinal environment by acting as decoy receptors for pathogens and as prebiotics, which promote the colonization of commensal bacteria. Here, using α2,3- and α2,6-sialyltransferase-deficient mice, we investigated the role of the sialylated milk oligosaccharides sialyl(α2,3)lactose and sialyl(α2,6)lactose on mucosal immunity. The exposure of newborn mice to milk containing or deficient in sialyllactose had no impact on the development of mucosal leukocyte populations. However, when challenged by dextran sulfate sodium (DSS) in drinking water, adult mice that had been fostered on sialyl(α2,3)lactose-deficient milk were more resistant to colitis compared with mice fostered on normal milk or sialyl(α2,6)lactose-deficient milk. Analysis of intestinal microbiota showed different colonization patterns depending on the presence or absence of sialyl(α2,3)lactose in the milk. Germ-free mice reconstituted with intestinal microbiota isolated from mice fed on sialyl(α2,3)lactose-deficient milk were more resistant to DSS-induced colitis than germ-free mice reconstituted with standard intestinal microbiota. Thus, exposure to sialyllactose during infancy affects bacterial colonization of the intestine, which influences the susceptibility to DSS-induced colitis in adult mice.


Chemical constituents and free radical scavenging activity of corn pollen collected from Apis mellifera hives compared to floral corn pollen at Nan, Thailand.
Bee pollen is composed of floral pollen mixed with nectar and bee secretion that is collected by foraging honey (Apis sp.) and stingless bees. It is rich in nutrients, such as sugars, proteins, lipids, vitamins and flavonoids, and has been ascribed antiproliferative, anti-allergenic, anti-angiogenic and free radical scavenging activities. This research aimed at a preliminary investigation of the chemical constituents and free radical scavenging activity in A. mellifera bee pollen. Bee pollen was directly collected from A. mellifera colonies in Nan province, Thailand, in June, 2010, whilst floral corn (Zea mays L.) pollen was collected from the nearby corn fields. The pollen was then sequentially extracted with methanol, dichloromethane (DCM) and hexane, and each crude extract was tested for free radical scavenging activity using the DPPH assay, evaluating the percentage scavenging activity and the effective concentration at 50% (EC50). The most active crude fraction from the bee pollen was then further enriched for bioactive components by silica gel 60 quick and adsorption or Sephadex LH-20 size exclusion chromatography. The purity of all fractions in each step was observed by thin layer chromatography and the bioactivity assessed by the DPPH assay. The chemical structures of the most active fractions were analyzed by nuclear magnetic resonance. The crude DCM extract of both the bee corn pollen and floral corn pollen provided the highest active free radical scavenging activity of the three solvent extracts, but it was significantly (over 28-fold) higher in the bee corn pollen (EC(50) = 7.42 ± 0.12 μg/ml), than the floral corn pollen (EC(50) = 212 ± 13.6% μg/ml). After fractionation to homogeneity, the phenolic hydroquinone and the flavone 7-O-R-apigenin were found as the minor and major bioactive compounds, respectively. Bee corn pollen contained a reasonably diverse array of nutritional components, including biotin (56.7 μg/100 g), invert sugar (19.9 g/100 g), vitamin A and β carotene (1.53 mg/100 g). Bee pollen derived from corn (Z. mays), a non-toxic or edible plant, provided a better free radical scavenging activity than floral corn pollen.


[p SRM, SWATH and HRM - targeted proteomics approaches on TripleTOF 5600+ mass spectrometer and their applications in oncology research].
Development of novel diagnostic and therapeutic approaches in cancer research requires sensitive and quantitative assays for determination of cancer associated proteins in clinical samples. Novel quantitative targeted proteomic approaches are overviewed in this communication. A major advantage of selected reaction monitoring (SRM) and pseudo- SRM lies in the selective and sensitive quantification of selected proteins in large sample sets. As such, they represent an alternative to immunochemical approaches. On the other hand, the potential of HRM and SWATH lies in recording of digital fingerprints, which enable post acquisition quantitative proteomic data mining on a similar basis to SRM. This article shows applications of targeted proteomics in a number of cancer research studies where they were used for quantification and validation of current or potential protein bio-markers and to study their role in cancer development and progression.


Haemophilia.
The clinical and haematological features of haemophilia are reviewed and the historical aspects, pathogenesis and genetics are discussed. The problems associated with management, such as factor replacement, home therapy, physiotherapy, surgical intervention and the presence of inhibitors, are fully described. Finally, mention is made of the psychological problems of haemophiliacs and of the help given by the South African Haemophilia Foundation.


Infrared Chemical Nano-Imaging: Accessing Structure, Coupling, and Dynamics on Molecular Length Scales.
This Perspective highlights recent advances in infrared vibrational chemical nano-imaging. In its implementations of scattering scanning near-field optical microscopy (s-SNOM) and photothermal-induced resonance (PTIR), IR nanospectroscopy provides few-nanometer spatial resolution for the investigation of polymer, biomaterial, and related soft-matter surfaces and nanostructures. Broad-band IR s-SNOM with coherent laser and synchrotron sources allows for chemical recognition with small-ensemble sensitivity and the potential for sensitivity reaching the single-molecule limit. Probing selected vibrational marker resonances, it gives access to nanoscale chemical imaging of composition, domain morphologies, order/disorder, molecular orientation, or crystallographic phases. Local intra- and intermolecular coupling can be measured through frequency shifts of a vibrational marker in heterogeneous environments and associated inhomogeneities in vibrational dephasing. In combination with ultrafast spectroscopy, the vibrational coherent evolution of homogeneous sub-ensembles coupled to their environment can be observed. Outstanding challenges are discussed in terms of extensions to coherent and multidimensional spectroscopies, implementation in liquid and in situ environments, general sample limitations, and engineering s-SNOM scanning probes to better control the nano-localized optical excitation and to increase sensitivity.