Applying additional autologous platelet-rich fibrin matrix or serial platelet-rich plasma to microfracture technique increases the quality of the repaired cartilage.
The aim of the present study is to investigate and compare the effects of biological adjuvants (platelet-rich plasma, platelet-rich fibrin matrix) and microfracture technique individually and in combination on full thickness chondral defects in a rabbit model. A total of 60 New Zealand White rabbits were randomly divided into six groups according to treatment modality as follows: control (C), microfracture (MF), platelet-rich plasma (PRP), platelet-rich fibrin matrix (PRFM), platelet-rich fibrin matrix after microfracture (MF + PRFM) and platelet-rich plasma after microfracture (MF + PRP) groups. The cartilage repair tissue was assessed histologically via International Cartilage Repair Score (ICRS) and macroscopically via ICRS macroscopic assessment scale. It was shown that overall macroscopic scores of the groups with MF were higher than those of the groups without MF. The cell morphology observed in the defect areas was mostly characterized with non-chondrocyte cells in the groups without MF, whereas chondrocyte cells mostly prevailed in the groups with MF. There was a greater integration through the cartilage-like tissue in the MF + PRP and MF + PRFM groups. The control group showed either fissures or fissures partially filled with fibrous tissue. When the groups were individually examined, there were statistically significant differences between the control and MF groups (p = 0.002), between the control and MF + PRFM groups (p = 0.001), between the control and MF + PRP groups (p < 0.001), between the PRFM and MF + PRFM groups (p = 0.014) and between the PRFM and MF + PRP (p = 0.023) groups in terms of histological evaluation scores. The application of PRP and PRFM in combination with MF treatment exhibited a positive impact on the repair and restoration of cartilage, and produced better outcomes than the individual use of PRP and PRFM. Nevertheless, in the treatment of full thickness chondral defects, the use of PRFM injection is recommended, which is performed intraoperatively at a single time and with no difficulty of repeating after surgery, instead of serial PRP injections based on the macroscopic and histological results obtained in the present study indicating that there was no significant difference between the use of these two adjuvants.


Access to thalidomide for the treatment of multiple myeloma in Canada: physician behaviours and ethical implications.
Multiple myeloma is an incurable malignancy. Since the late 1990s, its management has changed with the introduction of novel agents. Thalidomide, which is often called a "novel" therapy, has significantly prolonged survival in multiple myeloma and is considered worldwide to be part of standard of care in this disease. However, thalidomide is not approved in Canada, leading to problems with drug access for patients. Our study surveyed Canadian hematologists on their thalidomide prescribing practices and difficulties with drug access. We address some of the ethical issues facing patients and their doctors who are unable to obtain or afford the drug, and who therefore resort to alternative means such as illegal importation. Of the 411 Canadian hematologists contacted, 122 completed the survey, 97 reported that they did not treat myeloma, and 192 did not respond. Assuming that all non-responders treat myeloma, our estimated overall response rate from physicians who treat this disease was 39%. Survey participants indicated that, in Canada, access to thalidomide is a major issue for physicians and myeloma patients alike, and that 81% of respondents are dissatisfied or very dissatisfied with the drug access process. Many physicians felt that the special access process for thalidomide is unduly onerous, influences treatment decisions, and invades patient privacy. We found that 20% of physicians were unaware of the legal implications of obtaining thalidomide from other countries and that at least 23% overtly or covertly support patients in obtaining the drug from a non-Health-Canada-approved source. The current lack of access to thalidomide in Canada is a concerning problem for patients and health care providers dealing with myeloma. Regulatory changes at the federal level (Health Canada) need to be re-examined to promptly resolve this issue.


The yin and yang of tumor necrosis factor inhibitors.
Tumor necrosis factor (TNF) inhibitors have proven highly effective against a number of autoimmune diseases but have been disappointing in treating others. An increase in the risk of Mycobacterium tuberculosis and other opportunistic infections has been noted in patients treated with these agents. If we use these drugs, we need to weigh their beneficial and adverse effects.


Improving conformational searches by geometric screening.
Conformational searches in molecular docking are a time-consuming process with wide range of applications. Favorable conformations of the ligands that successfully bind with receptors are sought to form stable ligand-receptor complexes. Usually a large number of conformations are generated and their binding energies are examined. We propose adding a geometric screening phase before an energy minimization procedure so that only conformations that geometrically fit in the binding site will be prompted for energy calculation. Geometric screening can drastically reduce the number of conformations to be examined from millions (or higher) to thousands (or lower). The method can also handle cases when there are more variables than geometric constraints. An early-stage implementation is able to finish the geometric filtering of conformations for molecules with up to nine variables in 1 min. To the best of our knowledge, this is the first time such results are reported deterministically. firstname.lastname@example.com.


COMPARE-AMI trial: comparison of intracoronary injection of CD133+ bone marrow stem cells to placebo in patients after acute myocardial infarction and left ventricular dysfunction: study rationale and design.
Stem cell therapy has emerged as a promising approach to improve healing of the infarcted myocardium, to treat or prevent cardiac failure, and to restore lost cardiac function. Despite initial excitement, recent clinical trials using nonhomogenous human stem cells preparations showed variable results, raising concerns about the best cell type to transplant. Selected CD133(+) hematopoietic stem cells are promising candidate cells with great potential. COMPARE-acute myocardial infarction (AMI) study is a phase II, randomized, double-blind, placebo-controlled trial evaluating the safety and effectiveness of intracoronary CD133(+)-enriched hematopoietic bone marrow stem cells in patients with acute myocardial infarction and persistent left ventricular dysfunction. Patients who underwent successful percutaneous coronary intervention and present a persistent left ventricular ejection fraction <50% will be eligible to have bone marrow aspiration and randomized for intracoronary injection of selected CD 133(+) bone marrow cells vs placebo. The primary end point is a composite of a safety and efficacy end points evaluating the change at 4 months in the coronary atherosclerotic burden progression proximal and distal to the coronary stent in the infarct related artery; and the change in global left ventricular ejection fraction at 4 months relative to baseline as measured by magnetic resonance imaging. The secondary end point will be the occurrence of a major adverse cardiac event. To date, 14 patients were successfully randomized and treated without any protocol-related complication. COMPARE-AMI trial will help identify the effect of a selected population of the bone marrow stem cells on cardiac recovery of infarcted myocardium.


Uncovering the In Vivo Proxisome Using Proximity-Tagging Methods.
The development of new approaches is critical to gain further insights into biological processes that cannot be obtained by existing methods or technologies. The detection of protein-protein interaction is often challenging, especially for weak and transient interactions or for membrane proteins. Over the last decade, several proximity-tagging methodologies have been developed to explore protein interactions in living cells. Among those, the most efficient are based on protein partner modification, such as biotinylation or pupylation. Such technologies are based on engineered variants of enzymes like peroxidases or ligases that release reactive molecules, in the presence of specific substrates, that bind surrounding proteins. Fusing a protein of interest (POI) to these enzymes allows the definition of an unbiased "proxisome," that is, all of the proteins in interaction or in close vicinity of the POI. Here, the different proximity-labeling tools available are described and comprehensive comparison to discuss advantages and limitations is provided.


Scientific misconduct and accountability in teams.
Increasing complexity and multidisciplinarity make collaboration essential for modern science. This, however, raises the question of how to assign accountability for scientific misconduct among larger teams of authors. Biomedical societies and science associations have put forward various sets of guidelines. Some state that all authors are jointly accountable for the integrity of the work. Others stipulate that authors are only accountable for their own contribution. Alternatively, there are guarantor type models that assign accountability to a single author. We contribute to this debate by analyzing the outcomes of 80 scientific misconduct investigations of biomedical scholars conducted by the U.S. Office of Research Integrity (ORI). We show that the position of authors on the byline of 184 publications involved in misconduct cases correlates with responsibility for the misconduct. Based on a series of binary regression models, we show that first authors are 38% more likely to be responsible for scientific misconduct than authors listed in the middle of the byline (p<0.01). Corresponding authors are 14% more likely (p<0.05). These findings suggest that a guarantor-like model where first authors are ex-ante accountable for misconduct is highly likely to not miss catching the author responsible, while not afflicting too many bystanders.


DockStream: a docking wrapper to enhance de novo molecular design.
Recently, we have released the de novo design platform REINVENT in version 2.0. This improved and extended iteration supports far more features and scoring function components, which allows bespoke and tailor-made protocols to maximize impact in small molecule drug discovery projects. A major obstacle of generative models is producing active compounds, in which predictive (QSAR) models have been applied to enrich target activity. However, QSAR models are inherently limited by their applicability domains. To overcome these limitations, we introduce a structure-based scoring component for REINVENT. DockStream is a flexible, stand-alone molecular docking wrapper that provides access to a collection of ligand embedders and docking backends. Using the benchmarking and analysis workflow provided in DockStream, execution and subsequent analysis of a variety of docking configurations can be automated. Docking algorithms vary greatly in performance depending on the target and the benchmarking and analysis workflow provides a streamlined solution to identifying productive docking configurations. We show that an informative docking configuration can inform the REINVENT agent to optimize towards improving docking scores using public data. With docking activated, REINVENT is able to retain key interactions in the binding site, discard molecules which do not fit the binding cavity, harness unused (sub-)pockets, and improve overall performance in the scaffold-hopping scenario. The code is freely available at https://github.com/MolecularAI/DockStream .


Risk factors for venous thromboembolism in pediatric trauma.
Venous thromboembolism (VTE) is a major source of morbidity in critically ill trauma patients. Although the incidence and risk factors for VTE after trauma in adults have been well described, similar data regarding pediatric patients are lacking. Pediatric (age < 16 years) trauma patients with VTE were identified from a large administrative database collated from 19 states across the United States. Risk factors for VTE were identified using multivariate techniques. Risk of VTE increased with age and Injury Severity Scores. VTE was clearly associated with head, thoracic, abdominal, lower extremity, and spinal injuries. Craniotomy, laparotomy, and spinal operations were also associated with VTE. The greatest risk of VTE was in children with venous catheters. Older children with high Injury Severity Scores, major vascular injury, craniotomy, or venous catheters are at risk for VTE. These data may help guide strategies geared toward screening and prophylaxis in injured children.


A systematic review of differences in outcome between one and two stage palate repair in cleft lip and palate.
The aim of this systematic review was to determine whether one-stage palatoplasty for children born with cleft lip and palate shows overall advantages in outcome compared with two-stage palatoplasty. The included studies were controlled studies of syndromic and non-syndromic children born with unilateral cleft lip and palate, bilateral cleft lip and palate, or isolated cleft palate. The interventions studied were one-stage palatoplasty and two-stage palatoplasty starting with the soft palate. The outcomes were facial growth, speech, hearing, presence of fistulae, other complications related to surgery, health-related quality of life, and health economics. In total, 14 original studies were included. Results were dichotomized into showing advantage for one- or two-stage palatoplasty for the respective outcome and compared with the results from six included systematic reviews. No overall advantage for either surgical strategy was found for any of the outcome measures. The certainty of evidence was highest for the presence of fistulae, followed by facial growth and speech. For several outcomes, the quality of the existing evidence was too low to allow for any conclusions to be drawn. Neither one- nor two-stage palatoplasty showed significant advantages in clinical outcomes compared with the other. Other aspects such as ethics, economics, or surgeon's preference might hence be of more importance. Homogenous choices of outcome measures and defined minimal clinically important differences would facilitate further research.


Quantifying abortion rates of reproductive organs and effects of contributing factors using time-to-event analysis.
Time-to-event analysis, or survival analysis, is a method to analyse the timing of events and to quantify the effects of contributing factors. We apply this method to data on the timing of abortion of reproductive organs. This abortion often depends on source and sink strength. We hypothesise that the effect of source and sink strength on abortion rate can be quantified with a statistical model, obtained via survival analysis. Flower and fruit abortion in Capsicum annuum L., observed in temperature and planting density experiments, were analysed. Increasing the source strength as well as decreasing the sink strength decreased the abortion rate. The effect was non-linear, e.g. source strengths above 6g CH2O per plant per d did not decrease abortion rates further. The maximum abortion rate occurred around 100 degree-days after anthesis. Analyses in which sink strength was replaced with the number of fruits in a specified age category had an equal or better fit to the data. We discuss the advantages and disadvantages of using survival analyses for this kind of data. The technique can also be used for other crops showing reproductive organ abortion (e.g. soybean (Glycine max L.), cucumber (Cucumis sativus L.)), but also on other event types like bud break or germination.


Multistate Outbreak of Salmonella Virchow Infections Linked to a Powdered Meal Replacement Product-United States, 2015-2016.
Nontyphoidal Salmonella is the leading cause of bacterial gastroenteritis in the United States. Meal replacement products containing raw and "superfood" ingredients have gained increasing popularity among consumers in recent years. In January 2016, we investigated a multistate outbreak of infections with a novel strain of Salmonella Virchow. Cases were defined using molecular subtyping procedures. Commonly reported exposures were compared with responses from healthy people interviewed in the 2006-2007 FoodNet Population Survey. Firm inspections and product traceback and testing were performed. Thirty-five cases from 24 states were identified; 6 hospitalizations and no deaths were reported. Thirty-one of 33 (94%) ill people interviewed reported consuming a powdered supplement in the week before illness; of these, 30 (97%) reported consuming product A, a raw organic powdered shake product consumed as a meal replacement. Laboratory testing isolated the outbreak strain of Salmonella Virchow from leftover product A collected from ill people's homes, organic moringa leaf powder (an ingredient in product A), and finished product retained by the firm. Firm inspections at 3 facilities linked to product A production did not reveal contamination at the facilities. Traceback investigation identified that the contaminated moringa leaf powder was imported from South Africa. This investigation identified a novel outbreak vehicle and highlighted the potential risk with similar products not intended to be cooked by consumers before consuming. The company issued a voluntary recall of all implicated products. As this product has a long shelf life, the recall likely prevented additional illnesses.


Association of antithrombin with development of trauma-induced disseminated intravascular coagulation and outcomes.
Trauma activates the innate immune system to modulate hemostasis and minimize the damage caused by physiological bodily responses, including the activation of coagulation. Sufficiently severe trauma overwhelms physiological responses and elicits the systemic inflammatory response syndrome, which leads to the onset of disseminated intravascular coagulation (DIC), characterized by dysregulated inflammatory coagulofibrinolytic responses. Impaired anticoagulant mechanisms, including antithrombin, constitutes the pathology of DIC, while the dynamics of antithrombin and relevance to outcomes in trauma-induced coagulopathy have not been fully elucidated. This study investigated the associations of antithrombin activity with DIC onset and outcomes in severely injured patients. This retrospective sub-analysis of a multicenter, prospective study included patients with an injury severity score ≥16. We characterized trauma patients with low antithrombin activity (antithrombin <80% on hospital arrival, n = 75) in comparison with those who had normal antithrombin activity (antithrombin ≥80%, n = 200). Global markers of coagulation and fibrinolysis, molecular biomarkers for thrombin generation (soluble fibrin [SF]), and markers of anticoagulation (antithrombin) were evaluated to confirm the associations of antithrombin with DIC development and outcomes, including in-hospital mortality and the multiple organ dysfunction syndrome (MODS). Patients with low antithrombin activity had higher prevalence of shock, transfusion requirements, and in-hospital mortality. Higher DIC scores and more severe organ dysfunction were observed in the low AT group compared to that in the normal AT group. Antithrombin activity on arrival at the hospital was an independent predictor of the development of DIC in trauma patients, and levels of SF increased with lower antithrombin values (antithrombin activity > 85%). Antithrombin activity at 3 h showed good predictive performance for in-hospital mortality, and a multivariable Cox proportional-hazard regression model with a cross-product term between the antithrombin and DIC showed that the in-hospital mortality in patients with DIC increased with decreased antithrombin activity. A multivariable logistic regression model showed that the odds for the development of MODS in patients with DIC increased with lower antithrombin values. Decreased antithrombin activity in trauma-induced coagulopathy is associated with poor outcomes through worsening of DIC.


Hematopoietic growth-factor signal transduction and regulation of gene expression.
Hemopoietic cells have an absolute requirement for survival and proliferation for specific growth factors. The growth factors maintain the critical vitality of the cells by stimulating adenosine triphosphate (ATP) synthesis and hexose transport. Intracellular alkalinization also occurs rapidly through the stimulation of the Na+/H+ antiporter. These immediate metabolic events, not initiated by serum components, appear to be necessary for the integrity of cellular viability (Fig. 6). Interleukin-3 has been shown to induce the activation of PK-C through a mechanism(s) not requiring the hydrolysis of phosphoinositol 4,5 bisphosphate. A role for Ca2+ influx or intracellular release in the action of CSFs or interleukins has not been shown. Although downregulation of cAMP has been reported in response to IL-2, the signal transduction process of CSFs and IL-2 appears not to be mediated by upregulation of cyclic nucleotide metabolism or "classical" phospholipid degradative pathways. Protein phosphorylation is clearly modulated by the hemopoietic cytokines, yet only the CSF-1 receptor has any known intrinsic kinase activity. Instead, the IL-3, GM-CSF receptors, and perhaps G-CSF appear to be coupling to kinases of both tyrosine and serine specificities. This may be a direct allosteric interaction with membrane-associated kinases or transduced through an intermediate protein such as those using GTP. Such is the case for many hormone receptors that couple to amplifying "second messenger" enzyme systems (i.e., adenylate cyclase, phospholipase C) or members of the insulin growth factor family that couple to tyrosine kinases in proximity to the receptors (IGF-II). One of the kinase systems that IL-2, IL-3, and other CSFs stimulate appears to have some characteristics similar to PK-C. Direct activators of PK-C stimulate some similar serine-threonine phosphorylation and perhaps even tyrosine phosphorylation. The hemopoietic growth factors, however, stimulate tyrosine phosphorylation of some proteins that are not phosphorylated in response to PK-C activators, suggesting that these kinase systems are independently regulated. Although phorbol esters stimulate many of the same metabolic activities (ATP synthesis in myeloid and lymphoid cell lines), growth-factor abrogation is clearly associated with the action of tyrosine kinase oncogenes or the nuclear oncogene effectors such as v-myc. It is likely, therefore, that tyrosine kinases are playing a critical role in the control of proliferation although the dominant amount of cellular protein phosphorylations are on serine. Both classes of kinases are apparently required for growth-factor action. All the hemopoietic growth factors examined thus far stimulate the steady-state accumulation of the nuclear protooncogenes.(ABSTRACT TRUNCATED AT 400 WORDS)


Simple model of spiking neurons.
A model is presented that reproduces spiking and bursting behavior of known types of cortical neurons. The model combines the biologically plausibility of Hodgkin-Huxley-type dynamics and the computational efficiency of integrate-and-fire neurons. Using this model, one can simulate tens of thousands of spiking cortical neurons in real time (1 ms resolution) using a desktop PC.


Correlative 3D x-ray fluorescence and ptychographic tomography of frozen-hydrated green algae.
Accurate knowledge of elemental distributions within biological organisms is critical for understanding their cellular roles. The ability to couple this knowledge with overall cellular architecture in three dimensions (3D) deepens our understanding of cellular chemistry. Using a whole, frozen-hydrated Chlamydomonas reinhardtii cell as an example, we report the development of 3D correlative microscopy through a combination of simultaneous cryogenic x-ray ptychography and x-ray fluorescence microscopy. By taking advantage of a recently developed tomographic reconstruction algorithm, termed GENeralized Fourier Iterative REconstruction (GENFIRE), we produce high-quality 3D maps of the unlabeled alga's cellular ultrastructure and elemental distributions within the cell. We demonstrate GENFIRE's ability to outperform conventional tomography algorithms and to further improve the reconstruction quality by refining the experimentally intended tomographic angles. As this method continues to advance with brighter coherent light sources and more efficient data handling, we expect correlative 3D x-ray fluorescence and ptychographic tomography to be a powerful tool for probing a wide range of frozen-hydrated biological specimens, ranging from small prokaryotes such as bacteria, algae, and parasites to large eukaryotes such as mammalian cells, with applications that include understanding cellular responses to environmental stimuli and cell-to-cell interactions.


Cabomba as a model for studies of early angiosperm evolution.
The angiosperms, or flowering plants, diversified in the Cretaceous to dominate almost all terrestrial environments. Molecular phylogenetic studies indicate that the orders Amborellales, Nymphaeales and Austrobaileyales, collectively termed the ANA grade, diverged as separate lineages from a remaining angiosperm clade at a very early stage in flowering plant evolution. By comparing these early diverging lineages, it is possible to infer the possible morphology and ecology of the last common ancestor of the extant angiosperms, and this analysis can now be extended to try to deduce the developmental mechanisms that were present in early flowering plants. However, not all species in the ANA grade form convenient molecular-genetic models. The present study reviews the genus Cabomba (Nymphaeales), which shows a range of features that make it potentially useful as a genetic model. We focus on characters that have probably been conserved since the last common ancestor of the extant flowering plants. To facilitate the use of Cabomba as a molecular model, we describe methods for its cultivation to flowering in the laboratory, a novel Cabomba flower expressed sequence tag database, a well-adapted in situ hybridization protocol and a measurement of the nuclear genome size of C. caroliniana. We discuss the features required for species to become tractable models, and discuss the relative merits of Cabomba and other ANA-grade angiosperms in molecular-genetic studies aimed at understanding the origin of the flowering plants.


Administration of a soluble activin type IIB receptor promotes the transplantation of human myoblasts in dystrophic mice.
Duchenne muscular dystrophy (DMD) is a recessive disease caused by a dystrophin gene mutation. Myoblast transplantation permits the introduction of the dystrophin gene into dystrophic muscle fibers. However, this strategy has so far produced limited results. Modulation of transforming growth factor-β (TGF-β) superfamily signaling promotes skeletal muscle differentiation and growth and myogenic regeneration. We investigated the possibility that the combination of TGF-β superfamily signaling inhibition with myoblast transplantation might be an effective therapeutic approach in dystrophin-deficient patients. In vitro, blocking myostatin and other ligands with a soluble form of the extracellular domain of the activin IIB receptor (ActRIIB/Fc) upregulated the expression of myogenic differentiation factors and increased human myoblast fusion. In vivo, systemic inhibition of activin IIB receptor signaling by delivery of ActRIIB/Fc increased the success of the myoblast transplantation. This effect was further increased by forcing the mice to swim weekly to induce cycles of muscle degeneration and regeneration. Treatment of dystrophic mice with ActRIIB/Fc led to increased body weight, increased skeletal muscle mass, and improved myoblast transplantation. Thus, ActRIIB/Fc represents an effective therapeutic strategy for muscular dystrophies, and its effects are enhanced when combined with muscle exercise.


Mais Médicos (More Doctors) Program: its contribution in view of WHO recommendations for provision of doctors.
In order to examine whether Brazil's Mais Médicos (More Doctors) Programme (PMM) reflected World Health Organisation (WHO) recommendations for improved attraction, retention and recruitment of health workers in remote and rural areas, this descriptive, qualitative study drew on document analysis in order to compare the WHO recommendations published in 2010 with Brazil's Law No. 12,871/13, which instituted the PMM. Of the 16 WHO recommendations systematised here, the PMM met 37.5%. Recommendations not incorporated into the PMM include career development programmes and public recognition strategies. Although reflecting WHO recommendations and already in place elsewhere in the SUS prior to announcement of the PMM, the National Retention Grant Programme and multi-professional teams (as in the Family Health Strategy) were not implemented by the PMM. The programme contains innovative components such as a new curriculum for medical schools and compulsory medical service. On the other hand, the PMM could have invested more in personal and professional support.


Expression of growth/differentiation factor 11, a new member of the BMP/TGFbeta superfamily during mouse embryogenesis.
We have cloned and characterized a new member of the bone morphogenetic protein/transforming growth factor beta (BMP/TGFbeta) superfamily, growth differentiation factor 11 (Gdf11), from rat incisor pulp RNA by reverse transcription-polymerase chain reaction using degenerate primers. The mature carboxyl-terminal domain encoded by Gdf11 is most closely related to Gdf8, being 90% identical to the mouse gene. Northern blot analysis revealed Gdf11 is expressed in adult dental pulp and brain. In situ hybridization of sections and whole-mount embryos demonstrated Gdf11 is first strongly expressed in restricted domains at 8.5 days post coitus (dpc) when it is highest in the tail bud. At 10.5 dpc, it is expressed in the branchial arches, limb bud, tail bud and posterior dorsal neural tube. Later, it is expressed in terminally-differentiated odontoblasts, the nasal epithelium, retina and specific regions of the brain.


Increased belowground biomass and soil CO2 fluxes after a decade of carbon dioxide enrichment in a warm-temperate forest.
Atmospheric CO2 concentrations have risen 40% since the start of the industrial revolution. Beginning in 1996, the Duke Free-Air CO2 Enrichment experiment has exposed plots in a loblolly pine forest to an additional 200 microL/L CO2 compared to trees growing in ambient CO2. This paper presents new belowground data and a synthesis of results through 2008, including root biomass and nutrient concentrations, soil respiration rates, soil pore-space CO2 concentrations, and soil-solution chemistry to 2 m depth. On average in elevated CO2, fine-root biomass in the top 15 cm of soil increased by 24%, or 59 g/m2 (26 g/m2 C). Coarse-root biomass sampled in 2008 was twice as great in elevated CO2 and suggests a storage of approximately 20 g C x m(-2) x yr(-1). Root C and N concentrations were unchanged, suggesting greater belowground plant demand for N in high CO2. Soil respiration was significantly higher by 23% on average as assessed by instantaneous infrared gas analysis and 24-h integrated estimates. N fertilization decreased soil respiration and fine-root biomass by approximately 10-20% in both ambient and elevated CO2. In recent years, increases in root biomass and soil respiration grew stronger, averaging approximately 30% at high CO2. Peak changes for root biomass, soil respiration, and other variables typically occurred in midsummer and diminished in winter. Soil CO2 concentrations between 15 and 100 cm depths increased 36-60% in elevated CO2. Differences from 30 cm depth and below were still increasing after 10 years' exposure to elevated CO2, with soil CO2 concentrations >10000 microL/L higher at 70- and 100-cm depths, potentially influencing soil acidity and rates of weathering. Soil solution Ca2+ and total base cation concentrations were 140% and 176% greater, respectively, in elevated CO2 at 200 cm depth. Similar increases were observed for soil-solution conductivity and alkalinity at 200 cm in elevated CO2. Overall, the effect of elevated CO2 belowground shows no sign of diminishing after more than a decade of CO2 enrichment.


The determination of resectability of lung cancer by fiberoptic bronchoscopy.
The usefulness of flexible fiberoptic bronchoscopy (FFB) in determining resectability of lung cancer in 152 patients was evaluated. Based on clinical and routine chest roentgenographic examinations, 66 patients (43%) had what were considered to be operable lesions greater than 3 cm in diameter or tumors associated with atelectasis or obstructive pneumonitis (but less than an entire lung), without evidence of pleural effusion or chest wall, mediastinal, or extrathoracic involvement. Forty-eight (73%) of the 66 patients were eventually found to have unresectable conditions. With FFB alone, the conditions of 32 (67%) of these 48 patients were staged as unresectable, so that more invasive procedures, eg, mediastinoscopy or thoracotomy, were required in only 12 and four patients, respectively. Early FFB should be done in potentially resectable cases in which these roentgenographic criteria are met. This procedure is easy, safe, and economical and would allow many patients with lung cancer to have the stage of their disease determined and be treated entirely as outpatients.


Problems in dealing with missing data and informative censoring in clinical trials.
Acommon problem in clinical trials is the missing data that occurs when patients do not complete the study and drop out without further measurements. Missing data cause the usual statistical analysis of complete or all available data to be subject to bias. There are no universally applicable methods for handling missing data. We recommend the following: (1) Report reasons for dropouts and proportions for each treatment group; (2) Conduct sensitivity analyses to encompass different scenarios of assumptions and discuss consistency or discrepancy among them; (3) Pay attention to minimize the chance of dropouts at the design stage and during trial monitoring; (4) Collect post-dropout data on the primary endpoints, if at all possible; and (5) Consider the dropout event itself an important endpoint in studies with many.


Fast fully 3-D image reconstruction in PET using planograms.
We present a method of performing fast and accurate three-dimensional (3-D) backprojection using only Fourier transform operations for line-integral data acquired by planar detector arrays in positron emission tomography. This approach is a 3-D extension of the two-dimensional (2-D) linogram technique of Edholm. By using a special choice of parameters to index a line of response (LOR) for a pair of planar detectors, rather than the conventional parameters used to index a LOR for a circular tomograph, all the LORs passing through a point in the field of view (FOV) lie on a 2-D plane in the four-dimensional (4-D) data space. Thus, backprojection of all the LORs passing through a point in the FOV corresponds to integration of a 2-D plane through the 4-D "planogram." The key step is that the integration along a set of parallel 2-D planes through the planogram, that is, backprojection of a plane of points, can be replaced by a 2-D section through the origin of the 4-D Fourier transform of the data. Backprojection can be performed as a sequence of Fourier transform operations, for faster implementation. In addition, we derive the central-section theorem for planogram format data, and also derive a reconstruction filter for both backprojection-filtering and filtered-backprojection reconstruction algorithms. With software-based Fourier transform calculations we provide preliminary comparisons of planogram backprojection to standard 3-D backprojection and demonstrate a reduction in computation time by a factor of approximately 15.


Squalene does not exhibit a chemopreventive activity and increases plasma cholesterol in a Wistar rat hepatocarcinogenesis model.
The eventual chemopreventive effect of squalene (SQ), a triterpene present in olive oil, was evaluated when administered to Wistar rats during a period comprising the initiation and selection/promotion of the "resistant hepatocyte" (RH) model of hepatocarcinogenesis. During 8 consecutive wk, animals received by gavage SQ (100 or 150 mg/100 g body weight) dissolved in corn oil (CO) daily. Animals treated with only CO and submitted to the RH model were used as controls. Treatments with SQ did not result in inhibition of macroscopically visible hepatocyte nodules (P > 0.05) or of hepatic placental glutathione S-transferase- positive preneoplastic lesions (PNL; P > 0.05). Hepatic cell proliferation and apoptosis indexes were not different (P > 0.05) among the different experimental groups, both regarding PNL and surrounding normal tissue areas. There were no significant differences (P > 0.05) among comets presented by rats treated with the two SQ doses or with CO. On the other hand, SQ increased total plasma cholesterol levels when administered at both doses (P < 0.05). This indicates that the isoprenoid was absorbed. Thus, SQ did not present chemopreventive activity during hepatocarcinogenesis and had a hypercholesterolemic effect, suggesting caution when considering its use in chemoprevention of cancer.


Prevalence and Predicting Factors of Chronic Pain among Workers of Petrochemical and Petroleum Refinery Plants.
Taking into account the differences in job requirements and conditions, it is expected that workers in some occupations are more susceptible to pain than others. To examine the prevalence of chronic pain among workers of several petrochemical and petroleum refinery plants. We also examined the predictive role of psycho-familial variables (depression, work-family conflict and job stress) in causing chronic pain when controlling for demographic and occupational factors. This cross-sectional study was carried out among 674 workers. Those with chronic pain were identified by affirmative answers to screening questions based on the ICD-11 criteria. There were 162 (24.0%; 95% CI 20.8% to 27.3%) workers meeting the ICD- 11 criteria for chronic pain. Headache was the most frequently reported pain (29.9%). We found a significantly (p=0.03) higher prevalence of pain among the middle age than in other age groups. Chronic pain more frequently affected divorced/widowed workers (p<0.001), and those with more work experience (p=0.04). Workers with chronic pain reported significantly higher levels of depression (p<0.001), job stress (p=0.007), and work-family conflict (p<0.001). After controlling for demographic and occupational factors, depression (p<0.001) and work-family conflict (p=0.003) were found to be independent predictors of chronic pain among studied workers. Workers who experience higher levels of depression, work-family conflict and job stress might be more prone to chronic pain. The majority of these factors are modifiable, and the problem may thus be solved by establishing appropriate screening programs, and availability of proper services and education.


Activity and tissue-specific expression of the transcription factor NF-E1 multigene family.
NF-E1, a DNA-binding protein that recognizes the general consensus motif WGATAR, is the first tissue-specific factor to be identified in erythroid cells. Using a probe from the murine GF-1 (NF-E1) cDNA clone, we isolated three homologous chicken cDNAs: One of these corresponds to an mRNA (NF-E1a) that is abundantly and exclusively expressed in erythroid cells; a second mRNA (NF-E1b) is also expressed in all developmental stages of erythroid cells but is additionally found in a limited subset of other chicken tissues; mRNA representative of a third gene (NF-E1c) is expressed only in definitive (adult) erythrocytes within the red cell lineage but is also abundantly expressed in T lymphocytes and brain. All NF-E1 proteins are highly conserved within the DNA-binding domain and bind to the consensus motif with similar affinities in vitro; they are also all stimulatory trans-acting factors in vivo. The factors differ quantitatively in their ability to trans-activate reporter genes in which the number and position of cognate binding sites is varied relative to the transcriptional initiation site. These data suggest that the NF-E1 consensus motif directs a broader and more complicated array of developmental transcriptional regulatory processes than has been assumed and that NF-E1c may play a unique regulatory role in the developing chicken brain and in T lymphocytes.


Which patient benefit most from minimally invasive direct anterior approach total hip arthroplasty in terms of perioperative blood loss? A retrospective comparative study from a cohort of patients with primary degenerative hips.
Total hip arthroplasty (THA) is a successful surgery, but despite the advancements in anesthesiology and orthopedics, sometimes blood transfusions are required to manage the anemia due to the blood loss, involving a substantial number of patients. The aim of this retrospective comparative study is to define how the choice of the surgical approach, either direct anterior (DA) or posterolateral (PL), may influence the postoperative blood loss and the need for transfusion in THA. Data collection was carried out retrospectively of THAs performed between 2016 and 2021 on primary hip osteoarthritis treated by DA or with PL approach. Clinical and perioperative anesthetic data were collected. Preoperative hemoglobin levels were compared with the lowest detected level by calculating ΔHb (hemoglobin decrease). Then, data from the two groups were cross-checked: duration of surgery, whether premedication with tranexamic acid, duration of the hospitalization, rate of need for hemotransfusions, and amount of blood transfused. The two samples were subdivided into subgroups according to age, BMI, tranexamic acid prophylaxis, and chronic treatment with drugs that alter coagulative properties. Time of surgery was longer for patients treated with DA access (mean DA: 78.8 min; mean PL: 74.8 min; p: 0.05; 95% CI), but the length of hospitalization was shorter for patients treated with DA group with a mean time of 6.23 days versus 7.12 days for the PL group (p < 0.01). DA THA resulted advantageous mainly in patients between 66 and 75 years, showing a reduced postoperative transfusion requirement in the postoperative period (DA: 13.43%-mean: 1.33 units; PL: 26.82%-mean: 1.18 units; p: 0.044, 95% CI). Patients that assume blood-altering drugs showed a higher transfusion rate (p < 0.01), but comparison of the two subgroups showed that the choice of the surgical approach did not significantly affect the transfusion rate in these patients (p: 0.512). Prophylaxis with tranexamic acid reduced the transfusion rate (p < 0.01). Patients treated by minimally invasive direct anterior approach undergo a significantly shorter hospitalization. From the analysis of patient's subgroups those aged between 66- and 75-years benefit from the DA approach mainly for the minor blood loss with less frequent transfusion requirement.


Dual-modality imaging system for monitoring human heart organoids beating in vitro.
To reveal the three-dimensional microstructure and calcium dynamics of human heart organoids (hHOs), we developed a dual-modality imaging system combining the advantages of optical coherence tomography (OCT) and fluorescence microscopy. OCT provides high-resolution volumetric structural information, while fluorescence imaging indicates the electrophysiology of the hHOs' beating behavior. We verified that concurrent OCT motion mode (M-mode) and calcium imaging retrieved the same beating pattern from the heart organoids. We further applied dynamic contrast OCT (DyC-OCT) analysis to strengthen the verification and localize the beating clusters inside the hHOs. This imaging platform provides a powerful tool for studying and assessing hHOs in vitro, with potential applications in disease modeling and drug screening.


Relationships among hope, psychological well-being and health-related quality of life in childhood cancer survivors.
This cross-sectional study examined the level of hope among Hong Kong childhood cancer survivors and investigated the relationships among hope, depressive symptoms, self-esteem and health-related quality of life. We recruited 176 survivors aged 10-16 years who underwent medical follow-ups at the outpatient clinic. This study revealed that lower levels of hope were associated with an increase in depressive symptoms and reductions in self-esteem and health-related quality of life. Our results contribute to novel findings by demonstrating that hope may be a significant factor associated with health-related quality of life. This understanding could increase healthcare professionals' awareness about the psychological needs of childhood cancer survivors.


CNN and SVM-Based Models for the Detection of Heart Failure Using Electrocardiogram Signals.
Heart failure (HF) is a serious condition in which the heart fails to supply the body with enough oxygen and nutrients to function normally. Early and accurate detection of heart failure is critical for impeding disease progression. An electrocardiogram (ECG) is a test that records the rhythm and electrical activity of the heart and is used to detect HF. It is used to look for irregularities in the heart's rhythm or electrical conduction, as well as a history of heart attacks, ischemia, and other conditions that may initiate HF. However, sometimes, it becomes difficult and time-consuming to interpret the ECG signal, even for a cardiac expert. This paper proposes two models to automatically detect HF from ECG signals: the first one introduces a Convolutional Neural Network (CNN), while the second one suggests an extension of it by integrating a Support Vector Machine (SVM) layer for the classification at the end of the network. The proposed models provide a more accurate automatic HF detection using 2-s ECG fragments. Both models are smaller than previously proposed models in the literature when the architecture is taken into account, reducing both training time and memory consumption. The MIT-BIH and the BIDMC databases are used for training and testing the adopted models. The experimental results demonstrate the effectiveness of the proposed framework by achieving an accuracy, sensitivity, and specificity of over 99% with blindfold cross-validation. The models proposed in this study can provide doctors with reliable references and can be used in portable devices to enable the real-time monitoring of patients.


Human lens epithelial cell proliferation in a protein-free medium.
The ocular humors are relatively low in protein, yet cell growth in the human capsular bag still occurs after extracapsular cataract extraction (ECCE) surgery. This resilient growth gives rise to posterior capsule opacification (PCO) in a significant proportion (30%) of patients. This study compared the ability of human lens cells to proliferate in serum-supplemented and protein-free medium. Sham cataract operations were performed on human donor eyes. The capsular bag was dissected free, pinned flat on a petri dish, and incubated in Eagle's minimal essential medium (EMEM) alone or in EMEM supplemented with 10% fetal calf serum. Observations were made by phase-contrast microscopy. At the endpoint, capsules were studied by fluorescence or electron microscopy. Mitotic activity was identified using Bromo-2-deoxyuridine labeling and detection techniques. When required, an intraocular lens was implanted when surgery was performed. It was found that human lens cells from a wide age spectrum of donors proliferate and migrate on the lens capsule in the absence of added protein. The rate of growth was age-dependent, such that the posterior capsule was completely confluent after 8.0 +/- 0 days (n = 3) and 24.4 +/- 5.3 days (n = 3) for donor lenses aged < 40 years and > 60 years, respectively. The outgrowth of epithelial cells gave rise to capsular contraction, wrinkling, and increased light scatter. Growth on the anterior surface of the intraocular lens was less prolific than on the posterior capsule. The protein-free model replicates many features of clinically-observed PCO. The resilient cell growth on the natural collagen capsule explains the high prevalence of PCO, especially in younger patients, and suggests that inflammation and external growth factors are not necessary for PCO. Furthermore, the protein-free capsular bag system can be used to explore fundamental questions concerning the autocrine control of lens epithelial cell survival and growth.


Investigating Global Lipidome Alterations with the Lipid Network Explorer.
Lipids play an important role in biological systems and have the potential to serve as biomarkers in medical applications. Advances in lipidomics allow identification of hundreds of lipid species from biological samples. However, a systems biological analysis of the lipidome, by incorporating pathway information remains challenging, leaving lipidomics behind compared to other omics disciplines. An especially uncharted territory is the integration of statistical and network-based approaches for studying global lipidome changes. Here we developed the Lipid Network Explorer (LINEX), a web-tool addressing this gap by providing a way to visualize and analyze functional lipid metabolic networks. It utilizes metabolic rules to match biochemically connected lipids on a species level and combine it with a statistical correlation and testing analysis. Researchers can customize the biochemical rules considered, to their tissue or organism specific analysis and easily share them. We demonstrate the benefits of combining network-based analyses with statistics using publicly available lipidomics data sets. LINEX facilitates a biochemical knowledge-based data analysis for lipidomics. It is availableas a web-application and as a publicly available docker container.


Elder women's perceptions around optimal perinatal health: a constructivist grounded-theory study with an Indigenous community in southern Ontario.
Women play important roles in translating health knowledge, particularly around pregnancy and birth, in Indigenous societies. We investigated elder Indigenous women's perceptions around optimal perinatal health. Using a methodological framework that integrated a constructivist grounded-theory approach with an Indigenous epistemology, we conducted and analyzed in-depth interviews and focus groups with women from the Six Nations community in southern Ontario who self-identified as grandmothers. Our purposive sampling strategy was guided by a Six Nations advisory group and included researcher participation in a variety of local gatherings as well as personalized invitations to specific women, either face-to-face or via telephone. Three focus groups and 7 individual interviews were conducted with 18 grandmothers. The participants' experiences converged on 3 primary beliefs: pregnancy is a natural phase, pregnancy is a sacred period for the woman and the unborn child, and the requirements of immunity, security (trust), comfort, social development and parental responsibility are necessary for optimal postnatal health. Participants also identified 6 communal responsibilities necessary for families to raise healthy children: access to healthy and safe food, assurance of strong social support networks for mothers, access to resources for postnatal support, increased opportunities for children to participate in physical activity, more teachings around the impact of maternal behaviours during pregnancy and more teachings around spirituality/positive thinking. We also worked with the Six Nations community on several integrated knowledge-translation elements, including collaboration with an Indigenous artist to develop a digital story (short film). Elder women are a trusted and knowledgeable group who are able to understand and incorporate multiple sources of knowledge and deliver it in culturally meaningful ways. Thus, tailoring public health programming to include elder women's voices may improve the impact and uptake of perinatal health information for Indigenous women.


Rapid carbene formation increases ion diffusivity in an imidazolium acetate ionic liquid confined between polar glass plates.
1-Ethyl-3-methyl-imidazolium acetate ([EMIM][OAc]) is one of the most widely used ionic liquids for various applications. This study is focussed on the chemical stability of [EMIM][OAc] on the surfaces of polar glass plates. 1H and 13C NMR spectroscopy and NMR diffusometry of [EMIM][OAc] IL confined between glass plates with a specific surface area 105-106 m-1 are thoroughly investigated. A rapid and spontaneous reaction took place on the surfaces of glass plates leading to the formation of neutral chemical moieties as evident by the appearance of new signals in the 1H NMR spectra. These new products are assigned as N-heterocyclic carbene (NHC) and acetic acid. These neutral chemical moieties have significantly increased the ion diffusivity by dissociation of the cation and the anion in [EMIM][OAc] IL. The yield and rate of formation of NHC and acetic acid are found to increase with the increasing surface area of polar glass plates and the time of contact between the IL and glass surfaces. Based on NMR spectroscopy, a dissociative reaction mechanism is proposed for the formation of free NHC in the neat [EMIM][OAc] IL.


Persistent expression of Notch2 delays gonadotrope differentiation.
Normal pituitary gland development requires coordination between maintenance of progenitor cell pools and selection of progenitors for differentiation. The spatial and temporal expression of Notch2 during pituitary development suggested that it could control progenitor cell differentiation in the pituitary. Consistent with this idea, Notch2 is not expressed in Prop1 mutants, and anterior pituitary progenitors in Prop1 mutants appear to be unable to transition from proliferation to differentiation properly, resulting in anterior lobe failed cell specification and evolving hypoplasia. To test the function of Notch2 directly, we used the alphaGSU subunit promoter to express activated NOTCH2 persistently in pre-gonadotropes and pre-thyrotropes of transgenic mice. At birth, there is a small reduction in the population of fully differentiated thyrotropes and almost no fully differentiated gonadotropes. The temporal and spatial expression of Hey1 suggests that it could be a mediator of this effect. Gonadotropes complete their differentiation program eventually, although expression of LH and FSH is mutually exclusive with NOTCH2 transgene expression. This demonstrates that activated Notch2 is sufficient to delay gonadotrope differentiation, and it supports the hypothesis that Notch2 regulates progenitor cell differentiation in the pituitary gland.


Reconstruction of ancestral haplotypes in a 12-generation schizophrenia pedigree.
We searched for candidate chromosomal regions inherited identical by descent in 19 patients suffering from schizophrenia or schizoaffective disorder that are related 12 generations back, to an ancestral couple born in the middle of the seventeenth century. To accomplish this goal, we constructed complete chromosomal haplotypes for each patient using genotype data from 450 markers. In total, 12 haplotype regions (with sizes ranging from 0.6 to 10.9 cM) constituted by three markers each were identical in three or more of the affected individuals. The largest genomic segment was located on 6q25, a region previously shown to be significantly more frequent in patients than controls, and proposed to contain a schizophrenia susceptibility locus. For the remaining 11 candidate haplotypes, we estimated haplotype frequencies from all the 43 affected members collected from the same family and 46 unrelated control individuals. This analysis indicated that at least four of the 11 candidate haplotypes are ancestral, since the frequencies were significantly higher in patients than in controls. Five additional haplotypes showed higher estimated frequencies in the patients but the differences were not significant. Interestingly, five of these 11 genomic regions are located in, or close to, candidate regions previously suggested to contain susceptibility genes for schizophrenia. The regions are 5q21-23, 8p21-22, 10p13-15, 13q12-13 and 22q12-13. Several of these haplotypes are probably ancestral linkage disequilibrium blocks inherited from the original couple. There exists, however, the possibility that one or more of these regions harbour schizophrenia susceptibility loci that may have epistatic interactions among them.


Communicating Value in Simulation: Cost-Benefit Analysis and Return on Investment.
Value-based health care requires a balancing of medical outcomes with economic value. Administrators need to understand both the clinical and the economic effects of potentially expensive simulation programs to rationalize the costs. Given the often-disparate priorities of clinical educators relative to health care administrators, justifying the value of simulation requires the use of economic analyses few physicians have been trained to conduct. Clinical educators need to be able to present thorough economic analyses demonstrating returns on investment and cost-effectiveness to effectively communicate with administrators. At the 2017 Academic Emergency Medicine Consensus Conference "Catalyzing System Change through Health Care Simulation: Systems, Competency, and Outcomes," our breakout session critically evaluated the cost-benefit and return on investment of simulation. In this paper we provide an overview of some of the economic tools that a clinician may use to present the value of simulation training to financial officers and other administrators in the economic terms they understand. We also define three themes as a call to action for research related to cost-benefit analysis in simulation as well as four specific research questions that will help guide educators and hospital leadership to make decisions on the value of simulation for their system or program.


Isolation of a Golgi-apparatus-enriched fraction from leukaemic cells.
1. A Golgi-apparatus-enriched fraction was isolated from acute leukaemic lymphoblasts of AKR mice by using an homogenate stabilized with 1 mM-glutaraldehyde. 2. The isolated fraction, which was shown morphologically to be enriched in dictyosomes, possessed between 44- and 76-fold increase in specific activity, compared with the tumour homogenate, of UDP-galactose-glycoprotein galactosyltransferase and between 3- and10.5-fold increase in relative specific activity of UDP-N-acetygalactosamine-polypeptide N-acetylgalactosaminyltransferase. 3. Plasma membranes isolated from the leukaemic lymphoblasts also possessed glycoprotein galactosyltransferase activity, though in contrast with Golgi-apparatus-enriched material had no detectable polypeptide N-acetygalactosaminyltransferase. 4. The difficulties associated with maintaining the morphological integrity of the Golgi apparatus in subcellular fractionation are discussed.


Immunochemical studies with soluble and mitochondrial pyruvate carboxylase activities from rat tissues.
1. Pyruvate carboxylase (EC 22.214.171.124), purified from rat liver mitochondria to a specific activity of 14 units/mg, was used for the preparation of antibodies in rabbits. 2. Tissue distribution studies showed that pyruvate carboxylase was present in all rat tissues that were tested, with considerable activities both in gluconeogenic tissues such as liver and kidney and in tissues with high rates of lipogenesis such as white adipose tissue, brown adipose tissue, adrenal gland and lactating mammary gland. 3. Immunochemical titration experiments with the specific antibodies showed no differences between the inactivation of pyruvate carboxylase from mitochondrial or soluble fractions of liver, kidney, mammary gland, brown adipose tissue or white adipose tissue. 4. The antibodies were relatively less effective in reactions against pyruvate carboxylase from sheep liver than against the enzyme from rat tissues. 5. Pyruvate carboxylase antibodies did not inactivate either propionyl-CoA carboxylase or acetyl-CoA carboxylase from rat liver. 6. It is concluded that pyruvate carboxylase in lipogenic tissues is similar antigenically to the enzyme in gluconeogenic tissues and that the soluble activities of pyruvate carboxylase detected in many rat tissues do not represent discrete enzymes but are the result of mitochondrial damage during tissue homogenization.


High levels of maternally transferred mercury do not affect reproductive output or embryonic survival of northern watersnakes (Nerodia sipedon).
Maternal transfer is an important exposure pathway for contaminants because it can directly influence offspring development. Few studies have examined maternal transfer of contaminants, such as mercury (Hg), in snakes, despite their abundance and high trophic position in many ecosystems where Hg is prevalent. The objectives of the present study were to determine if Hg is maternally transferred in northern watersnakes (Nerodia sipedon) and to evaluate the effects of maternal Hg on reproduction. The authors captured gravid female watersnakes (n = 31) along the South River in Waynesboro, Virginia, USA, where an extensive Hg-contamination gradient exists. The authors measured maternal tissue and litter Hg concentrations and, following birth, assessed (1) reproductive parameters (i.e., litter size and mass, neonate mass); (2) rates of infertility, death during development, stillbirths, malformations, and runts; and (3) the overall viability of offspring. Mercury concentrations in females were strongly and positively correlated with concentrations in litters, suggesting that N. sipedon maternally transfer Hg in proportion to their tissue residues. Maternal transfer resulted in high concentrations (up to 10.10 mg/kg dry wt total Hg) of Hg in offspring. The authors found little evidence of adverse effects of Hg on these measures of reproductive output and embryonic survival, suggesting that N. sipedon may be more tolerant of Hg than other vertebrate species. Given that this is the first study to examine the effects of maternally transferred contaminants in snakes and that the authors did not measure all reproductive endpoints, further research is needed to better understand factors that influence maternal transfer and associated sublethal effects on offspring.


Partial alignment, residual dipolar couplings and molecular symmetry in solution NMR.
Residual dipolar couplings (RDCs) and residual anisotropic chemical shifts (RACSs) are produced by the partial alignment of solution NMR samples. RDCs and RACSs yield high-resolution structural and dynamic information on the orientation of bonds and chemical groups in molecules. Many molecules form oligomers or have intrinsic symmetries, which may simplify the analysis of their partial alignment datasets. In this report, we explore the theory of partial alignment using an irreducible spherical representation, and we investigate the impact of molecular symmetry on the alignment of molecules. Though previous studies have reported simplified relationships on the partial alignment of molecules bearing different symmetry groups, we show that these simplified relationships may not be universal and only apply to a limited set of systems.


Post-PCV7 changes in colonizing pneumococcal serotypes in 16 Massachusetts communities, 2001 and 2004.
The introduction of heptavalent conjugate pneumococcal vaccine (PCV7) has raised concerns for replacement with nonvaccine serotypes in both invasive disease and asymptomatic carriage. Analysis of colonizing serotypes among healthy children in the community provides critical data on such changes. Nasopharyngeal specimens were obtained from children who were younger than 7 years during well-child or sick visits in primary care practices in 16 Massachusetts communities during 2001 and 2004. Susceptibility testing and serotyping were performed on isolated Streptococcus pneumoniae strains. Vaccination history with PCV7 was abstracted from the medical record. Among colonizing pneumococcal isolates, PCV7 serotypes decreased from 36% to 14%, and non-PCV7 serotypes increased from 34% to 55%. Overall carriage did not change (26% to 23%); neither did carriage of potentially cross-reactive serotypes (30% to 31%). The most common non-PCV7 serotypes were serotypes 11, 15, and 29. There was a substantial increase in penicillin nonsusceptibility from 8% to 25% in non-PCV7 serotypes; 35% were highly resistant to penicillin. Penicillin nonsusceptibility increased from 45% to 56% among PCV7 serotypes while remaining stable among PCV7 potentially cross-reactive strains (51% vs 54%). Pneumococcal colonization has changed after the introduction of PCV7, both in serotype distribution and in patterns of antibiotic resistance. The frequency of nonvaccine strains has increased, and the proportion of nonvaccine isolates that are not susceptible to penicillin has tripled. This shift toward increased carriage of nonvaccine serotypes warrants vigilance for changes in the epidemiology of invasive pneumococcal disease.


Antimicrobial activities of the essential oils of Origanum onites L., Origanum vulgare L. subspecies hirtum (Link) Ietswaart, Satureja thymbra L., and Thymus cilicicus Boiss. & Bal. growing wild in Turkey.
In the present study, four separate samples of Origanum onites L., three separate samples of Satureja thymbra L., Origanum vulgare L. ssp. hirtum (Link) Ietswaart, and Thymus cilicicus Boiss. & Bal. were collected from various regions of Mugla, Turkey. The essential oils of these plants were obtained by the hydrodistillation method. Antimicrobial activities of the essential oils against microorganisms, including multiple antibiotic-resistant bacteria, were investigated using the disc diffusion method. Different antibiotic discs were used for comparison to the inhibition zones. All the essential oils used in this study were very effective against Gram-positive and Gram-negative bacteria, which included multiple resistant strains, except Pseudomonas aeruginosa ATCC 27853 and Pseudomonas fluorescens MU 87. The essential oils of the O. onites, O. vulgare ssp. hirtum, and S. thymbra were especially very effective against the resistant strains such as Stenotrophomonas maltophilia MU 64, S. maltophilia MU 99, and Chryseomonas luteola MU 65. The maximum antimicrobial activity was observed with the essential oils of O. onites. The antimicrobial activities of the essential oils varied depending on the species, subspecies, or variety. In fact, the essential oils of some plants belonging to the same species that were collected from different locations showed different levels of antimicrobial activities.


Methacholine bronchial responsiveness and variations in lung function among workers exposed to flour.
Methacholine bronchial responsiveness and variations in the pulmonary function of workers exposed to wheat flour and a reference group were compared. Each subject [140 men exposed to flour (bakers and pastry makers) and 77 controls] completed a standardized questionnaire. Bronchial responsiveness was quantified by measuring the slope between percentage decrements in forced expiratory volume in 1 second (FEV1) and cumulative methacholine dose. FEV1 and peak expiratory flow (PEF) were recorded four times a day for 15 days using a handheld electronic spirometer. The variability in the FEV1 and PEF readings was expressed as variation coefficients (100 x standard deviation/mean). The mean duration of exposure to flour was 14 (SD 9) years. Rhinitis was significantly more common in the exposed group than in the control group (30.7% versus 11.7%, P = 0.001). The mean FEV1 and PEF did not significantly differ between the two groups. The slope of the dose-response to methacholine and the variation coefficients were lower among the unexposed nonsmokers than among the exposed workers and smokers. The differences were significant for the exposed smokers. The two variation coefficients correlated with each other (r = 0.82) but not with the slope of the methacholine challenge. Conclusions Occupational exposure to flour and smoking increase bronchial responsiveness, as measured by the slope of the dose-response to methacholine and the variation coefficients of airflow. However, methacholine bronchial responsiveness and the variability of lung function do not measure exactly the same aspect of airway behavior.


Vaccine confidence among people who use drugs: A cross-sectional survey.
Adult immunization coverage remains low in the US, particularly for people who use drugs (PWUD), a population that experiences a disproportionate burden of vaccine-preventable diseases. The extent of and characteristics associated with vaccine confidence (VC) held by PWUD is poorly understood. As VC strongly correlates with vaccine uptake, this cross-sectional study identifies mutable factors associated with VC and quantifies its relationship to immunization status within a highly vulnerable, underimmunized population of PWUD. Using a community-engaged research strategy with select partner organizations hosting syringe exchange programs in Atlanta, Los Angeles, and Las Vegas, USA, we surveyed participants ages 18-69 years served by these organizations from 2019 to 2020. Survey measures included sociodemographics, health behavior including immunization receipt, and vaccine confidence in adult vaccinations using a modified Emory Vaccine Confidence Index (EVCI). The findings reflect relatively low VC among the 1,127 recruited participants, with 56% expressing low VC (EVCI 0-12), 35% medium (EVCI 13-20) and 10% high (EVCI 21-24). EVCI varied by city, with lowest confidence in Atlanta and highest in Las Vegas. VC was associated with past receipt of specific vaccines, including hepatitis A, MMR, Tdap, and influenza. VC varied by specific sociodemographic correlates such as housing insecurity (reduced confidence) and receipt of public benefits or disability (increased confidence). This study identified correlates associated with VC based on site and sociodemographic characteristics for this priority population, highlighting the need for specific interventions to raise VC among PWUD, especially among those experiencing housing insecurity and without public benefits.


Risk of severe periodontal disease in a Swedish adult population. A cross-sectional study.
In this study, potential risk factors for severe periodontal disease were identified in a cross-sectional sample from the county of Jonkoping, Sweden. 547 adults 20-70 years of age were categorised clinically and radiographically by level of periodontal disease experience. These levels were used to divide the sample into groups--individuals without any reduction in periodontal bone level (60%) and those with severe periodontal bone loss (13%)--which were then used in univariate and multivariate logistic regression analyses as dependent variable. Demographic, socio-economic, general health, smoking habits, clinical, and dental care variables were used in the different regression analyses. In the univariate model, age (20-70 years) was found to be correlated with more severe periodontal disease experience (odds ratio: 1.13; 95% CI: 1.10-1.17). The association with periodontal disease was more pronounced for the older age groups (50, 60, and 70 years). A negative financial situation was also related to severe periodontal bone loss when regressed univariately (odds ratio 2.20 [95%: 1.04-4.68]). Moderate-heavy smoking (> or =10 cigarettes/day) appeared to be associated with severe periodontal destruction with an odds ratio of 9.78 (95% CI: 3.62-36.42). Of the clinical variables in the univariate model, higher mean levels of supragingival dental plaque and the presence of subgingival calculus were related to more severe periodontal disease with odds ratios of 1.02 (95%: 1.01-1.03) and 2.96 (95%: 1.50-5.88), respectively. When the same variables were regressed multivariately, age (continuous) (odds ratio 1.17 [95% CI: 1.12-1.22]), moderate-heavy smoking (odds ratio 11.84 [95% CI: 4.19-33.50]), and higher mean levels of plaque (odds ratio 1.02 [95% CI: 1.00-1.03]) remained significant. Light smoking (1-9 cigarettes/day) was not significantly associated with severe periodontal disease in the 2 regression models. The present study demonstrated that smoking, greater age, and higher mean levels of plaque are potential risk factors for severe periodontal disease in this specific population.


The effects of function in fracture healing and stability.
A mechanically based classification system and hypothesis for fracture healing with peripheral callus has been proposed based on many years of clinical experience and numerous laboratory research studies. This hypothesis describes the interactions and influences on fracture healing from the vascular, mechanical, electric, chemical, and thermal environments in specific callus regions surrounding the fracture site. The central theme to this phenomenon is motion at the fracture site. The single factor distinguishing the effects of closed functional bracing of fractures from all other treatment modalities is motion between the fragments which results from early functional activity. Perhaps the stimulus to a prolonged inflammatory response is provided by friction between moving fracture fragments and the surrounding tissues. An increased vascular response results, and environmental factors are stimulated to form an abundant peripheral callus. Three distinct callus zones exist which provide overlapping stages of healing based on structure and optimized physiologic conditions. Clinical signs and symptoms, such as fracture stability as well as motion and pain at the fracture site, combine to provide the optimum feedback mechanism to control functional activity, which in turn governs environmental factors. Roentgenographic appearance can be better judged for mechanical significance if these three callus regions are appreciated. In animals as well as in humans, if function has correctly influenced the environment during callus formation, refracture strength is often greater than that of the original bone.


Resources and nutrients oriented greywater treatment for non-potable reuses.
This paper evaluated the performance and suitability of a resources and nutrients oriented decentralized greywater treatment system which uses a submerged spiral wound module. This greywater treatment system is aimed at treating and recovering the resources present in the wastewater. The study revealed that the UF membrane filtration system was able to maintain a permeate flux between 6 and 10 L/m2/h. TOC can be reduced from the influent value of 161 to 28.6 mg/L in the permeate, meaning an average elimination rate of 83.4%. In addition, soluble nutrients such as ammonia and phosphorus can pass through the UF membrane and remain in the permeate. The total nitrogen and total phosphorus in the permeate were 16.7 and 6.7 mg/L respectively. The permeate was low in turbidity (below 1 NTU) and free of suspended solids and E. coli and had an excellent physical appearance. The permeate can be used in gardening and agriculture for irrigation and soil fertilization or alternatively for toilet flushing after disinfection. The retentate generated in this system can be treated with blackwater and kitchen waste in an anaerobic digester at a later stage for producing biogas or compost.


Vascular access training for REBOA placement: a feasibility study in a live tissue-simulator hybrid porcine model.
The use of resuscitative endovascular balloon occlusion of the aorta (REBOA) in patients with severe haemorrhagic shock is increasing. Obtaining vascular access is a necessary prerequisite for REBOA placement in these situations. During the EVTM workshop (September 2017, Örebro, Sweden), 21 individuals participated in this study, 16 participants and five instructors. A formalised curriculum was constructed including basic anatomy of the femoral region and basic training in access materials for REBOA placement in zone 1. Key skills: (1) preparation of endovascular toolkit, (2) achieving vascular access in the model and (3) bleeding control with REBOA. Scoring ranged from 0 to 5 for non-anatomical skills. Identification of anatomical structures was either sufficient (score=1) or insufficient (score=0). Five consultants performed a second identical procedure as a post test. Consultants had significantly better overall technical skills in comparison with residents (p=0.005), while understanding of surgical anatomy showed no difference. Procedure times differed significantly (p<0.01), with residents having a median procedure time of 3 min and 24 s, consultants 2:33 and instructors 1:09. This comprehensive training model using a live tissue-simulator hybrid porcine model can be used for femoral access and REBOA placement training in medical personnel with different prior training levels. Higher levels of training are associated with faster procedure times. Further research in open and percutaneous access training is necessary to simulate real-life situations. This training method can be used in a multistep training programme, in combination with realistic moulage and perfused cadaver models.


Effects of noradrenaline infused into the chick hypothalamus on thermoregulation below thermoneutrality.
1. Hypothermia induced by infusion of noradrenaline into the hypothalamus of 2-3 week old chicks, within their thermoneutral range, was considerably potentiated by lowering ambient temperature. 2. Noradrenaline-induced hypothermia was associated with reduced carbon dioxide elimination and reduced blood lactate concentrations whereas leg temperature, electromyographic activity, plasma NEFA and plasma glucose concentrations were increased. 3. Mechanisms postulated to explain the phenomenon are inhibitory and facilitatory effects of noradrenaline on some, but not all, heat production and heat loss mechanisms. Increased electromyographic activity after intrahypothalamic noradrenaline is assumed to be due to lack of effect of noradrenaline on spinal thermosensitive centres; increased plasma NEFA concentration may be due to inhibition of NEFA utilization.


The surface of virulent Treponema pallidum: resistance to antibody binding in the absence of complement and surface association of recombinant antigen 4D.
The binding of immunoglobulin G present in syphilitic immune rabbit serum, syphilitic human serum, and rabbit antiserum to purified recombinant Treponema pallidum antigen 4D by T. pallidum, Nichols strain, was studied by immunoelectron microscopy. Treponemes were incubated with antiserum under the conditions of the T. pallidum immobilization test, in which T. pallidum-specific antibody renders the organism nonmotile and avirulent only in the presence of complement after a 16-h incubation period in an anaerobic environment. Antibody was not demonstrable on the surface of T. pallidum incubated with nonimmune rabbit serum or normal human serum in the presence of complement. Similarly, in the absence of complement, little or no antibody was found on the treponemal surface after incubation with syphilitic immune rabbit serum, syphilitic human serum, or rabbit antiserum directed against the recombinant 4D antigen. The addition of complement to syphilitic immune rabbit serum, syphilitic human serum, and anti-4D antibody resulted in immobilization and the deposition of antibody on the entire surface of the immobilized organisms. These results corroborate earlier work by other investigators demonstrating the resistance of freshly isolated T. pallidum to antibody binding in a variety of serological tests. Detection of 4D antigen on the surface of immobilized T. pallidum strongly implies that the use of T. pallidum immobilization test conditions provides a means to demonstrate the association of individual surface antigens on virulent T. pallidum. The resistance of T. pallidum to antibody binding may be relevant to the pathogenesis of syphilis.


Energy cost of the Trondheim firefighter test for experienced firefighters.
The aim of this study was to measure aerobic demands of fire fighting activities including exercise in the heat. Twenty-two experienced firefighters performed the Trondheim test simulating fire fighting tasks including work in the heat. Maximal oxygen uptake (VO2 max), heart rate (HR) and ventilation were recorded continuously. Data were compared with results obtained during a treadmill test during which the participants were dressed as smoke divers. The participants completed physical parts of the Trondheim test in ˜12 min (range: 7.5-17.4). Time to complete the test was closely related to the participant's VO2 max. HR of ˜170 beats/min and pulmonary ventilation of ˜100 L/min were higher than at lactate threshold (LT) during laboratory tests. VO2 averaged over the test's physical part was 35 ± 7 ml/min/kg, which was at the same or below the level corresponding to the participants' LT. Physically fit participants completed the test faster than less fit participants. Slower and physically less fit participants consumed more air and used more oxygen than faster and physically more fit participants. The Trondheim test is physically demanding; it distinguishes physically fit and less fit participants.


Loss of Bax alters tumor spectrum and tumor numbers in ARF-deficient mice.
p19(ARF) is a key regulator of the p53-mediated apoptotic and tumor suppressor pathway. The proapoptotic Bax gene is a transcription target of p53, yet genetic studies in some animal models have suggested that Bax and p53 loss may cooperate in tumorigenesis. ARF-deficient mice are tumor prone, and to determine whether Bax loss could cooperate in the development of these tumors, we generated mice null for both ARF and Bax. The tumor latency of Bax+/+ARF-/-, Bax+/-ARF-/- and Bax-/-ARF-/- mice was similar with a mean survival of 48.9, 48.1, and 47.6 weeks, respectively. In Bax+/+ARF-/- mice, the predominant tumor type was B- and T-cell lymphoma followed by sarcomas and a lack of carcinomas. However, the frequency of lymphoma development dramatically decreased, whereas that of sarcomas and carcinomas increased, in a gene dosage-dependent manner in Bax+/-ARF-/- and Bax-/-ARF-/- mice. Furthermore, uncommon tumors of ARF-/- mice (osteosarcoma and hemangiosarcoma) were observed in Bax/ARF-double null mice, and tumor types not described previously in ARF-null mice (mixed germ cell tumor, Triton tumor, and histiocytic sarcoma) also developed in Bax-/-ARF-/- animals. Importantly, multiple primary malignant tumors of different lineage arose in 25% of the Bax-/-ARF-/- mice, whereas only one tumor type per animal was observed in Bax+/+ARF-null littermates. Finally, the wild-type Bax allele was retained in tumors arising in Bax+/-ARF-/- mice. Thus, Bax appears to function as a tumor modifier rather than as a classic tumor suppressor, and the combined loss of Bax and the ARF allows for the emergence of multiple malignant tumor types, an alteration of the tumor spectrum, and tumors not observed previously in ARF-null mice.


[Physical examination--tourniquet tests for varicose veins].
Physical examination of patients with varicose veins is time-consuming and subjective. Nowadays, it is frequently replaced by hand-held Doppler examination and duplex scanning. Adequate studies investigating the value of physical examination for varicose veins are lacking. It is unclear whether the tourniquet tests of Trendelenburg and Perthes are more accurate than other physical tests, such as the cough test or tap test. In selected patients, who are not obese, have clearly visible varicose veins and have not been treated before, physical examination seems accurate, especially for assessment of the long saphenous vein. When physical examination is inconclusive, as often is the case with recurrent varicose veins, suspected short saphenous vein or perforator incompetence, or adipose legs, additional Doppler or duplex testing is required for an accurate diagnosis.


Clustering of risk factors in the smoking habits of schoolchildren in Sousse, Tunisia.
In Tunisia, little is known about the association between tobacco use and other chronic disease risk factors. This is the case for both adults and children. It is important to know the characteristics of young smokers to facilitate the creation and implementation of future programs for tobacco prevention. The aim of this study was to determine the association between tobacco use and other lifestyle factors among schoolchildren in Tunisia. We conducted a 2009/2010 cross-sectional questionnaire survey of 4003 randomly selected school children aged 13 years old (7th and 9th grades) to evaluate their knowledge, attitudes towards, and beliefs about the three risk factors for chronic disease (unhealthy diet, physical inactivity, and tobacco use). Written informed consent was obtained from each child's parents who allowed their child to participate. The mean age of our sample was 13.36 ± 1.28 years. The proportions of daily smokers were 2.2% and 0.1% among boys and girls, respectively. The proportions of irregular smokers were 9.1% and 1.5% among boys and girls, respectively. In our population, 19.1% (n=767) had ever experimented to smoke, with 29.8% among boys and 9% among girls (p<0.001). The proportions of schoolchildren who reported daily participation in physical activity were different between smokers and nonsmokers with 17.7% and 11.5%, respectively (p=0.03). Concerning eating habits, there was no significant difference in the consumption of fruits and vegetables; however, smokers frequently ate more high fat foods and in fast food restaurants. Similar results were found while comparing regular smoking children with those who experimented but who never became hooked on smoking. This study and previous research suggest the importance of early intervention in adolescents on smoking and combing these efforts with interventions focusing on physical activity and dietary habits.


[Evaluation of social demographic aspect of life quality of coal extraction workers in Kouzbass enterprises].
For assessing influence of social factors on life quality of workers in coal extraction enterpirses of Kemerovo region, the authors used questionnaire SF-36. Life quality parameters of workers engaged into coal extraction in Kemerovo region appeared to lower with age from 20 to 64 years. Life quality parameters on scales of pain, physical functioning and general health are invertedly correlated with age and length of service in hazardous work conditions for coal extraction workers. Life quality of the miners is influenced by the following factors: marital status, educational level and income level of the workers.


Cigarette smoke-induced necroptosis and DAMP release trigger neutrophilic airway inflammation in mice.
Recent data indicate a role for airway epithelial necroptosis, a regulated form of necrosis, and the associated release of damage-associated molecular patterns (DAMPs) in the development of chronic obstructive pulmonary disease (COPD). DAMPs can activate pattern recognition receptors (PRRs), triggering innate immune responses. We hypothesized that cigarette smoke (CS)-induced epithelial necroptosis and DAMP release initiate airway inflammation in COPD. Human bronchial epithelial BEAS-2B cells were exposed to cigarette smoke extract (CSE), and necrotic cell death (membrane integrity by propidium iodide staining) and DAMP release (i.e., double-stranded DNA, high-mobility group box 1, heat shock protein 70, mitochondrial DNA, ATP) were analyzed. Subsequently, BEAS-2B cells were exposed to DAMP-containing supernatant of CS-induced necrotic cells, and the release of proinflammatory mediators [C-X-C motif ligand 8 (CXCL-8), IL-6] was evaluated. Furthermore, mice were exposed to CS in the presence and absence of the necroptosis inhibitor necrostatin-1, and levels of DAMPs and inflammatory cell numbers were determined in bronchoalveolar lavage fluid. CSE induced a significant increase in the percentage of necrotic cells and DAMP release in BEAS-2B cells. Stimulation of BEAS-2B cells with supernatant of CS-induced necrotic cells induced a significant increase in the release of CXCL8 and IL-6, in a myeloid differentiation primary response gene 88-dependent fashion. In mice, exposure of CS increased the levels of DAMPs and numbers of neutrophils in bronchoalveolar lavage fluid, which was statistically reduced upon treatment with necrostatin-1. Together, we showed that CS exposure induces necrosis of bronchial epithelial cells and subsequent DAMP release in vitro, inducing the production of proinflammatory cytokines. In vivo, CS exposure induces neutrophilic airway inflammation that is sensitive to necroptosis inhibition.


Modeling adaptive reversible lanes: A cellular automata approach.
Dealing with traffic congestion is one of the most pressing challenges for cities. Transport authorities have implemented several strategies to reduce traffic jams with varying degrees of success. The use of reversible lanes is a common approach to improve traffic congestion during rush hours. A reversible lane can change its direction during a time interval to the more congested direction. This strategy can improve traffic congestion in specific scenarios. Most reversible lanes in urban roads are fixed in time and number; however, traffic patterns in cities are highly variable and unpredictable due to this phenomenon's complex nature. Therefore, reversible lanes may not improve traffic flow under certain circumstances; moreover, they could worsen it because of traffic fluctuations. In this paper, we use cellular automata to model adaptive reversible lanes(aka dynamic reversible lanes). Adaptive reversible lanes can change their direction using real-time information to respond to traffic demand fluctuations. Using real traffic data, our model shows that adaptive reversible lanes can improve traffic flow up to 40% compared to conventional reversible lanes. Our results show that there are significant fluctuations in traffic flow even during rush hours, and thus cities would benefit from implementing adaptive reversible lanes.


[Bronchial asthma pathogenesis and genetic prognosis development].
The review is dedicated to an actual problem--genetic prognosis of risk of bronchial asthma development that is quite a complex aspect of studies from a methodological viewpoint. Bronchial asthma--heterogeneous disease by both etiology and clinical characteristics. At the same time genetic prognosis is based on the unity of pathogenetic mechanisms of development, though in immunological reactions that are the base of this disease, alternative variants are possible. The aim of this review is carrying out parallels between modern achievements in the field of deciphering trigger mechanisms of bronchial asthma pathogenesis and object of genetic studies based on these mechanisms. Among the examined conceptions--role of epithelial tissue in trigger mechanisms of bronchial asthma, variants of key role of immune system cells, first of all, T-helpers of various types for further development of inflammatory-effector reactions with damage characteristic for this disease. Compliance of contemporary approaches of genetic studies and novel concepts of bronchial asthma pathogenesis is shown.


Content validation study of nursing interventions intended to prevent cardiovascular events in diabetic patients.
To validate nursing interventions from the Nursing Interventions Classification for preventing cardiovascular events in outpatients with diabetes. Cardiovascular disease is the main cause of death among diabetic patients. Although nurses play an important role in preventing cardiovascular events in this population, their specific contribution is difficult to measure. Documenting nursing interventions using a standardised nursing system may provide nursing visibility. However, there are no studies that analysed which nursing interventions would be the most relevant, based on expert opinion, aimed at preventing cardiovascular events in diabetic patients. This is a content validation study, which was designed as a descriptive one. For this content validation study, we used the Fehring model. Nursing interventions were selected from the Nursing Intervention Classification. Experts were asked to analyse the degree of relevance of 225 nursing interventions regarding the prevention of cardiovascular events in diabetic patients, considering its title and definition. We considered nursing interventions which obtained weighted arithmetic averages >0·80 to be valid. Among 50 invited experts, 14 (71·4% female; 36·5 + 9·7 years) responded and agreed to participate in this study. They analysed 225 preselected nursing interventions, and validated 29. The validated nursing interventions included documentation as well as biological, social and spiritual issues. The validated nursing interventions encompass a wide range of nursing knowledge, and these results provide empirical evidence for further studies.


Suppressive subtractive hybridization as a tool for identifying genetic diversity in an environmental metagenome: the rumen as a model.
Molecular techniques previously used for genome comparisons of closely related bacterial species could prove extremely valuable for comparisons of complex microbial communities, or metagenomes. Our study aimed to determine the breadth and value of suppressive subtractive hybridization (SSH) in a pilot-scale analysis of metagenomic DNA from communities of microorganisms in the rumen. Suppressive subtractive hybridization was performed using total genomic DNA isolated from rumen fluid samples of two hay-fed steers, arbitrarily designated as tester or driver. Ninety-six subtraction DNA fragments from the tester metagenome were amplified, cloned and the DNA sequences were determined. Verification of the isolation of DNA fragments unique to the tester metagenome was accomplished through dot blot and Southern blot hybridizations. Tester-specific SSH fragments were found in 95 of 96 randomly selected clones. DNA sequences of subtraction fragments were analysed by computer assisted DNA and amino acid comparisons. Putative translations of 26 (32.1%) subtractive hybridization fragments exhibited significant similarity to Bacterial proteins, whereas 15 (18.5%) distinctive subtracted fragments had significant similarity to proteins from Archaea. The remainder of the subtractive hybridization fragments displayed no similarity to GenBank sequences. This metagenomic approach has exposed an unexpectedly large difference in Archaeal community structure between the rumen microbial populations of two steers fed identical diets and housed together. 16S rRNA dot blot hybridizations revealed similar proportions of Bacteria and Archaea in both rumen samples and suggest that the differences uncovered by SSH are the result of varying community structural composition. Our study demonstrates a novel approach to comparative analyses of environmental microbial communities through the use of SSH.


The Caenorhabditis elegans muscle specific serpin, SRP-3, neutralizes chymotrypsin-like serine peptidases.
Members of the intracellular serpin family may help regulate apoptosis, tumor progression, and metastasis. However, their in vivo functions in the context of a whole organism have not been easily defined. To better understand the biology of these serpins, we initiated a comparative genomics study using Caenorhabditis elegans as a model organism. Previous in silico analysis suggested that the C. elegans genome harbors nine serpin-like sequences bearing significant similarities to the human clade B intracellular serpins. However, only five genes appear to encode full-length serpins with intact reactive site loops. To determine if this was the case, we have cloned and expressed a putative inhibitory-type C. elegans serpin, srp-3. Analysis of SRP-3 inhibitory activity indicated that SRP-3 was a potent inhibitor of the serine peptidases, chymotrypsin and cathepsin G. Spatial and temporal expression studies using GFP and LacZ promoter fusions indicated that SRP-3 was expressed primarily in the anterior body wall muscles, suggesting that it may play a role in muscle cell homeostasis. Combined with previous studies showing that SRP-2 is an inhibitor of the serine peptidase, granzyme B, and lysosomal cysteine peptidases, these data suggested that C. elegans expressed at least two inhibitory-type serpins with nonoverlapping expression and inhibitory profiles. Moreover, the profiles of these clade L serpins in C. elegans share significant similarities with the profiles of clade B intracellular serpin members in higher vertebrates. This degree of conservation suggests that C. elegans should prove to be a valuable resource in the study of metazoan intracellular serpin function.