Approximation for the vibrational excitation of middle atmospheric NO by upwelling radiation.
A plane-parallel approximation using FASCODE to calculate upward and downward IR flux density and angle-averaged radiance is described and tested against results that account for planetary curvature. The method is further developed to calculate the excitation of NO(v = 1) due to absorption of upwelling radiation in the earth's atmosphere. The algorithm is applied to a standard set of atmospheric models, in addition to typical and extreme desert atmospheric models which are included to maximize the effect. The results of this work show that upwelling radiation typically contributes <2.5% to the total NO(v = 1) excitation in the midstratosphere rises to ~5% at 50 km and becomes increasingly significant at higher altitudes. It is shown that in the mesosphere excitation due to upwelling and solar radiation become important compared to the dominant processes, thermal collisions, and chemical excitation. An approximate technique utilizing meteorological data, namely, tropospheric temperature, pressure, and humidity profiles, is developed to estimate the excitation of NO(v = 1) in the middle atmosphere. This technique would facilitate the retrieval of the NO mixing ratio from earthlimb emission data, as might be obtained from a satellite-borne limb sounding experiment, since it could be used to approximate the contribution of upwelling radiation to the NO(v = 1) non-LTE vibrational temperature efficiently.


Early Detection of Adverse Drug Reactions in Social Health Networks: A Natural Language Processing Pipeline for Signal Detection.
Adverse drug reactions (ADRs) occur in nearly all patients on chemotherapy, causing morbidity and therapy disruptions. Detection of such ADRs is limited in clinical trials, which are underpowered to detect rare events. Early recognition of ADRs in the postmarketing phase could substantially reduce morbidity and decrease societal costs. Internet community health forums provide a mechanism for individuals to discuss real-time health concerns and can enable computational detection of ADRs. The goal of this study is to identify cutaneous ADR signals in social health networks and compare the frequency and timing of these ADRs to clinical reports in the literature. We present a natural language processing-based, ADR signal-generation pipeline based on patient posts on Internet social health networks. We identified user posts from the Inspire health forums related to two chemotherapy classes: erlotinib, an epidermal growth factor receptor inhibitor, and nivolumab and pembrolizumab, immune checkpoint inhibitors. We extracted mentions of ADRs from unstructured content of patient posts. We then performed population-level association analyses and time-to-detection analyses. Our system detected cutaneous ADRs from patient reports with high precision (0.90) and at frequencies comparable to those documented in the literature but an average of 7 months ahead of their literature reporting. Known ADRs were associated with higher proportional reporting ratios compared to negative controls, demonstrating the robustness of our analyses. Our named entity recognition system achieved a 0.738 microaveraged F-measure in detecting ADR entities, not limited to cutaneous ADRs, in health forum posts. Additionally, we discovered the novel ADR of hypohidrosis reported by 23 patients in erlotinib-related posts; this ADR was absent from 15 years of literature on this medication and we recently reported the finding in a clinical oncology journal. Several hundred million patients report health concerns in social health networks, yet this information is markedly underutilized for pharmacosurveillance. We demonstrated the ability of a natural language processing-based signal-generation pipeline to accurately detect patient reports of ADRs months in advance of literature reporting and the robustness of statistical analyses to validate system detections. Our findings suggest the important contributions that social health network data can play in contributing to more comprehensive and timely pharmacovigilance.


Permeant cations modulate pore dynamics and gating of TRPV1 ion channels.
The transient receptor vanilloid 1 (TRPV1) is a non-selective ion channel, which is activated by several chemical ligands and heat. We have previously shown that activation of TRPV1 by different ligands results in single-channel openings with different conductance, suggesting that the selectivity filter is highly dynamic. TRPV1 is weakly voltage dependent; here, we sought to explore whether the permeation of different monovalent ions could influence the voltage dependence of this ion channel. By using single-channel recordings, we show that TRPV1 channels undergo rapid transitions to closed states that are directly connected to the open state, which may result from structural fluctuations of their selectivity filter. Moreover, we demonstrate that the rates of these transitions are influenced by the permeant ion, suggesting that ion permeation regulates the voltage dependence of these channels. Our data could be the basis for more detailed MD simulations exploring the permeation mechanism and how the occupancy of different ions alters the three-dimensional structure of the pore of TRPV1 channels.


Homodimerization of the glucocorticoid receptor is not essential for response element binding: activation of the phenylethanolamine N-methyltransferase gene by dimerization-defective mutants.
We have tested the commonly held hypothesis that glucocorticoid receptors (GRs) must dimerize via their DNA binding domain (DBD) to bind to glucocorticoid response elements (GRE) and induce gene expression. Guided by the GR dimerization-deficient dim/dim knock-in mouse, which expresses normal mRNA levels of the strictly GR-dependent phenylethanolamine N-methyltransferase (PNMT) gene, we analyzed in detail the regulation of the PNMT 5'-flanking region using wild-type GR (GRwt) and GR dimer mutants (GRdms). We demonstrated that mouse and rat PNMT 5'-regulatory fragments are more strongly induced by GRdms than by GRwt. Footprinting analysis revealed five regions where a GR-DBD peptide could bind. We delineated a 105-bp region containing two footprints with near-consensus glucocorticoid response elements and multiple half-sites that was sufficient for transactivation via both GRwt and GRdms. Finally, we demonstrated direct binding of GRdms proteins to this responsive region using EMSA. We propose that on a subset of GR-responsive promoters, exemplified by the PNMT gene, GRs can form concerted multimers in a manner that is independent of the DBD-dimer interface. We further suggest that protein-DNA and protein-protein interactions that support such complexes are essential for activation of this type of gene, and that DNA binding of GR might be essential to survival.


Developing content for a process-of-care checklist for use in intensive care units: a dual-method approach to establishing construct validity.
In the intensive care unit (ICU), checklists can be used to support the delivery of quality and consistent clinical care. While studies have reported important benefits for clinical checklists in this context, lack of formal validity testing in the literature prompted the study aim; to develop relevant 'process-of-care' checklist statements, using rigorously applied and reported methods that were clear, concise and reflective of the current evidence base. These statements will be sufficiently instructive for use by physicians during ICU clinical rounds. A dual-method approach was utilized; semi-structured interviews with local clinicians; and rounds of surveys to an expert Delphi panel. The interviews helped determine checklist item inclusion/exclusion prior to the first round Delphi survey. The panel for the modified-Delphi technique consisted of local intensivists and a state-wide ICU quality committee. Minimum standards for consensus agreement were set prior to the distribution of questionnaires, and rounds of surveys continued until consensus was achieved. A number of important issues such as overlap with other initiatives were identified in interviews with clinicians and integrated into the Delphi questionnaire, but no additional checklist items were suggested, demonstrating adequate checklist coverage sourced from the literature. These items were verified by local clinicians as being relevant to ICU and important elements of care that required checking during ward rounds. Two rounds of Delphi surveys were required to reach consensus on nine checklist statements: nutrition, pain management, sedation, deep vein thrombosis and stress ulcer prevention, head-of-bed elevation, blood glucose levels, readiness to extubate, and medications. Statements were developed as the most clear, concise, evidence-informed and instructive statements for use during clinical rounds in an ICU. Initial evidence in support of the checklist's construct validity was established prior to further prospective evaluation in the same ICU.


Control limits to identify outlying hospitals based on risk-stratification.
Outcome indicators are routinely used to compare hospitals with respect to quality of care. Indicators might be based on observed proportions of adverse events (binary outcomes) or observed averages of e.g. lengths or costs of hospital stays (continuous outcomes). These observed values are compared with expected ones in an average hospital, which might be estimated from a reference sample and should be appropriately adjusted for the case mix of patients. One possibility to achieve a reliable adjustment is to stratify the patients according to their risks, where each patient belongs to one and only one stratum. Control limits calculated under the null hypothesis of an average hospital, allowing to decide whether a discrepancy between an observed and an expected value might be explained by chance or not, are then plotted around the indicator, such that hospitals falling above those control limits are detected as being statistically worse than an average hospital. Calculation of valid control limits is however not always obvious. In this article, we propose a simple and unified framework to calculate such control limits when adjustment is based on stratification, where we allow to distinguish and disentangle the variability explained by stratification and the variability due to chance, where we take into account the uncertainty about the estimation of the expected values, and where it is possible not only to detect those hospitals which are statistically worse, but also those which are statistically much worse than an average hospital. The method applies both to binary and continuous outcomes and is illustrated on Swiss hospital discharge data.


[Amino acid assimilation by representatives of the Acholeplasma genus].
Amino acid assimilation by different representatives of Acholeplasma genus has been investigated. It was shown that all 7 investigated typical strains Acholeplasma laidlawii PG-8, A. granularum BTS-39, A. modicum PG-49, A. oculi 19-L, A. morum 72-043, A. axanthum S-743, A. hippikon C-1 and 37 phytopathogenic strains isolated from the affected plants were able to assimilate asparagine, glutamine, threonine, histidine and proline. A majority of acholeplasmas were able to metabolize phenylalanine, methionine, glutamate and lysine, rarely isoleucine. Each of the investigated species of acholeplasmas had individual spectrum of assimilated amino acids that can be used as an additional systematic characteristic of mollicutes.


Epigenetic control of cancer by neuropeptides.
Neuropeptides act as neurohormones, neurotransmitters and/or neuromodulators. Neuropeptides maintain physiological homeostasis and are paramount in molecular mechanisms of disease progression and regulation, including in cancer. Neuropeptides, by their definition, originate and are secreted from the neuronal cells, they are able to signal to neighboring cells or are released into the blood flow, if they act as neurohormones. The majority of neuropeptides exert their functions through G protein-coupled receptors, with certain exceptions. Although previous studies indicate that neuropeptides function in supporting proliferation of malignant cells in many types of solid tumor, the antitumorigenic action of the neuropeptides and their receptors, for example, in gastric cancers and chondrosarcoma, were also reported. It is known that epigenetically modified chromatin regulates molecular mechanisms involved in gene expression and malignant progression. The epigenetic modifications are genetically heritable, although they do not cause changes in DNA sequence. DNA methylation, histone modifications and miRNA expression are subject to those modifications. While there is substantial data on epigenetic regulation of neuropeptides, the epigenetic control of cancer by neuropeptides is considered to be uncharted territory. The aim of the current review is to describe the involvement of neuropeptides in the epigenetic machinery of cancer based on data obtained from our laboratory and from other authors.


Arthroscopic subacromial decompression for chronic impingement. Two- to five-year results.
Subacromial decompression was performed arthroscopically on 65 patients who were evaluated two to five years after the procedure. None had full thickness rotator cuff tears. Patients with partial thickness cuff tears were included in this study in order to allow comparison of arthroscopic acromioplasty with open acromioplasty for stage II impingement. On the UCLA shoulder rating scale, 89% of the cases in this study achieved a satisfactory result. These results compare favourably with those reported following open acromioplasty. The arthroscopic procedure is technically demanding. When properly performed in patients with appropriate indications, hospitalisation is brief, return to activities is rapid, there is little risk of deltoid muscle complications, and the results are lasting.


Genome editing in clinical genetics: points to consider-a statement of the American College of Medical Genetics and Genomics.
Disclaimer: These recommendations are designed primarily as an educational resource for medical geneticists and other health-care providers, to help them provide quality medical genetic services. Adherence to these recommendations does not necessarily assure a successful medical outcome. These recommendations should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results. In determining the propriety of any specific procedure or test, the geneticist should apply his or her own professional judgment to the specific clinical circumstances presented by the individual patient or specimen. It may be prudent, however, to document in the patient's record the rationale for any significant deviation from these recommendations.Genet Med advance online publication 26 January 2017.


An improved hybrid of particle swarm optimization and the gravitational search algorithm to produce a kinetic parameter estimation of aspartate biochemical pathways.
Mathematical modelling is fundamental to understand the dynamic behavior and regulation of the biochemical metabolisms and pathways that are found in biological systems. Pathways are used to describe complex processes that involve many parameters. It is important to have an accurate and complete set of parameters that describe the characteristics of a given model. However, measuring these parameters is typically difficult and even impossible in some cases. Furthermore, the experimental data are often incomplete and also suffer from experimental noise. These shortcomings make it challenging to identify the best-fit parameters that can represent the actual biological processes involved in biological systems. Computational approaches are required to estimate these parameters. The estimation is converted into multimodal optimization problems that require a global optimization algorithm that can avoid local solutions. These local solutions can lead to a bad fit when calibrating with a model. Although the model itself can potentially match a set of experimental data, a high-performance estimation algorithm is required to improve the quality of the solutions. This paper describes an improved hybrid of particle swarm optimization and the gravitational search algorithm (IPSOGSA) to improve the efficiency of a global optimum (the best set of kinetic parameter values) search. The findings suggest that the proposed algorithm is capable of narrowing down the search space by exploiting the feasible solution areas. Hence, the proposed algorithm is able to achieve a near-optimal set of parameters at a fast convergence speed. The proposed algorithm was tested and evaluated based on two aspartate pathways that were obtained from the BioModels Database. The results show that the proposed algorithm outperformed other standard optimization algorithms in terms of accuracy and near-optimal kinetic parameter estimation. Nevertheless, the proposed algorithm is only expected to work well in small scale systems. In addition, the results of this study can be used to estimate kinetic parameter values in the stage of model selection for different experimental conditions.


Genetic labeling does not detect epithelial-to-mesenchymal transition of cholangiocytes in liver fibrosis in mice.
Chronic injury changes the fate of certain cellular populations, inducing epithelial cells to generate fibroblasts by epithelial-to-mesenchymal transition (EMT) and mesenchymal cells to generate epithelial cells by mesenchymal-to-epithelial transition (MET). Although contribution of EMT/MET to embryogenesis, renal fibrosis, and lung fibrosis is well documented, role of EMT/MET in liver fibrosis is unclear. We determined whether cytokeratin-19 positive (K19(+)) cholangiocytes give rise to myofibroblasts (EMT) and/or whether glial fibrillary acidic protein positive (GFAP(+)) hepatic stellate cells (HSCs) can express epithelial markers (MET) in response to experimental liver injury. EMT was studied with Cre-loxP system to map cell fate of K19(+) cholangiocytes in K19(YFP) or fibroblast-specific protein-1 (FSP-1)(YFP) mice, generated by crossing tamoxifen-inducible K19(CreERT) mice or FSP-1(Cre) mice with Rosa26(f/f-YFP) mice. MET of GFAP(+) HSCs was studied in GFAP(GFP) mice. Mice were subjected to bile duct ligation or CCl(4)-liver injury, and livers were analyzed for expression of mesodermal and epithelial markers. On Cre-loxP recombination, >40% of genetically labeled K19(+) cholangiocytes expressed yellow fluorescent protein (YFP). All mice developed liver fibrosis. However, specific immunostaining of K19(YFP) cholangiocytes showed no expression of EMT markers alpha-smooth muscle actin, desmin, or FSP-1. Moreover, cells genetically labeled by FSP-1(YFP) expression did not coexpress cholangiocyte markers K19 or E-cadherin. Genetically labeled GFAP(GFP) HSCs did not express epithelial or liver progenitor markers in response to liver injury. EMT of cholangiocytes identified by genetic labeling does not contribute to hepatic fibrosis in mice. Likewise, GFAP(Cre)-labeled HSCs showed no coexpression of epithelial markers, providing no evidence for MET in HSCs in response to fibrogenic liver injury.


The Impact of Body Contouring Procedures on Urologic Outcomes in Massive Weight Loss Patients.
Post-massive weight loss body contouring is a growing area of plastic surgery. Studies have shown preexisting urinary incontinence amelioration in patients undergoing abdominal body-contouring procedures. These studies are small, focus on cosmetic abdominoplasties, and lack use of standardized surveys. The purpose of this study was to evaluate urinary incontinence endpoints in massive weight loss patients undergoing body-contouring procedures. A retrospective review was conducted over a 6-year period. Patients excluded had previous body-contouring procedures or previously treated urinary incontinence. Participants completed validated surveys to catalogue preoperative and postoperative urinary habits. Patients were then subdivided based on presence of preoperative incontinence. Outcomes were compared within and between cohorts using the t test and chi-square test. A total of 102 patients completed the survey. Of those, 44 had preoperative urinary incontinence. Patients with incontinence were found to be significantly older than those who did not. Postoperatively, patients with preoperative incontinence had significant decreases in incidence and severity of symptoms (p < 0.01, respectively), and significantly improved their quality of life (p < 0.02). Over 20 percent noted symptom resolution, and 67 percent were mostly or completely satisfied with the outcome. This study is the first to document amelioration of urinary incontinence symptoms in massive weight loss patients undergoing body-contouring procedures. It adds to the literature suggesting that abdominoplasties in select patients may improve incontinence symptoms. Future work will focus on evaluating the mechanism of this outcome and documenting improvement in an objective, prospective manner. Therapeutic, IV.


Achieving a 3-Star Society of Thoracic Surgery Lobectomy Ranking by Using Continuing Process Improvement, Lean Methodology, and Root Cause Analysis.
Our purpose is to identify the metrics used by the Society of Thoracic Surgeons (STS) to rank lobectomy and to show our process to improve. This is a review of our STS data for lobectomy and our results using the process of root cause analysis and lean methodology to improve our outcomes. The STS metrics are 30-day mortality, pneumonia, adult respiratory distress syndrome, bronchopleural fistula, pulmonary embolus, initial ventilator support greater than 48 hours, reintubation and respiratory failure, tracheostomy, myocardial infarction, or unexpected return to the operating room. Sixteen of 231 programs (7%) were ranked 3-star over a 3-year period from July 2011 to June 2014. The most common root cause analysis was failure to escalate care. The lean and process improvements we employed that seemed to improve the results were increasing exercise before surgery, adding a respiratory therapist, eliminating Foley catheters and arterial lines to reduce infection and to increase ambulation, offering stereotactic radiotherapy for marginal patients, favoring left upper segmentectomy over left upper lobectomy, and performing 91% of the last 493 lobectomies via a minimally invasive platform. Our major morbidity complications from August 2003 to December 2014 fell from 9.5% to 5.3% (P = 0.001) and mortality decreased from 3.3% to 0.54% (P < 0.0001). The metrics the STS used to rank lobectomy programs are 30-day mortality and predominantly respiratory complications. Root cause analysis, lean methodology, and process improvements allowed us to improve our lobectomy patient outcomes over time and to achieve a 3-star ranking over a 3-year time frame. These results may be obtainable by others.


Dietary Intake patterns in women with GDM and Non-GDM: A comparative study.
The study aimed to determine dietary Intake patterns in women with GDM and Non-GDM, a comparative study in a tertiary care hospital, Pakistan. This comparative cross sectional study was conducted through questionnaire spread over a period of six months of pregnant women visiting to Abbasi Shaheed Hospital for ante-natal visit having 24 to 28 weeks of gestation. With the written consent of the participants dietary intake patterns were assessed in GDM & Non-GDM subjects by a three day 24 hours' recalls and food frequency questionnaire. A 24-hour dietary recall chart is a dietary assessment tool in which participants were asked to recall all food and drink they have consumed in the last 24 hours. The FFQ (food frequency questionnaire) provide a list of foods and participants were asked how often they eat each item on the list. This FFQ has 70 food items. The food frequency was reported as never, per year, per month, once a week, once and a day. The reported intake of food was converted into nutrients intake (carbohydrate, protein, fat) which was calculated by reported intake frequency of each food multiplied by reported portion size and its respective nutrient composition, summing over all foods by a trained Nutritionist. A total of 75 participants with GDM, and 75 with Non-GDM were enrolled in this study over a period of six months. It was observed that dietary intake patterns have a significant association with GDM. Those who consume carbohydrate mainly containing diet have likely to have GDM. It has been seen that those who have family history of diabetes are more likely to have GDM. Family dietary patterns can affect risk of GDM. Our study has shown that timings of meals did not find have any significant association with GDM. Dietary patterns strongly influence the risk of GDM. The most contributing factors to risk of GDM are higher intake of carbohydrate rich diet and lesser consumption of fruits and vegetables.


An Overview on Wood Waste Valorization as Biopolymers and Biocomposites: Definition, Classification, Production, Properties and Applications.
Bio-based polymers, obtained from natural biomass, are nowadays considered good candidates for the replacement of traditional fossil-derived plastics. The need for substituting traditional synthetic plastics is mainly driven by many concerns about their detrimental effects on the environment and human health. The most innovative way to produce bioplastics involves the use of raw materials derived from wastes. Raw materials are of vital importance for human and animal health and due to their economic and environmental benefits. Among these, wood waste is gaining popularity as an innovative raw material for biopolymer manufacturing. On the other hand, the use of wastes as a source to produce biopolymers and biocomposites is still under development and the processing methods are currently being studied in order to reach a high reproducibility and thus increase the yield of production. This study therefore aimed to cover the current developments in the classification, manufacturing, performances and fields of application of bio-based polymers, especially focusing on wood waste sources. The work was carried out using both a descriptive and an analytical methodology: first, a description of the state of art as it exists at present was reported, then the available information was analyzed to make a critical evaluation of the results. A second way to employ wood scraps involves their use as bio-reinforcements for composites; therefore, the increase in the mechanical response obtained by the addition of wood waste in different bio-based matrices was explored in this work. Results showed an increase in Young's modulus up to 9 GPa for wood-reinforced PLA and up to 6 GPa for wood-reinforced PHA.


[Localization of minor proteins and structural changes in the myosin filaments of vertebrate striated muscle].
The origin of meridional reflections in the X-ray diffraction patterns of vertebrate skeletal muscles in resting and rigor states was studied. The main results may be summarized as follows. 1. Most of the meridional reflections localized in groups at the positions of successive orders of the repeat period of about 430 A are contributed mainly by the C-protein component of thick filaments. 2. The meridional reflections at about 143 and 72 A in the X-ray diffraction pattern of the resting muscle are contributed mainly by the cross-bridge axial repeat period, while in the X-ray diffraction patterns of the rigorized muscle the reflections at approximately the same positions are contributed mainly by C-protein. The change in the positions of these particular reflections accompanying the transition of the muscle from rest to rigor and from rest to contraction cannot be considered as an indication of a change in the axial repeat period of the cross-bridges, as it was earlier suggested by some authors. 3. The transition of the muscle from resting to rigor state is accompanied by substantial changes in the positions of the meridional reflections contributed my minor proteins, which is indicative of the structural transition in the thick filaments. The observed changes may be interpreted as the result of the thick filaments elongation by about 1.5% or, alternatively, as a consequence of the redistribution of electron density of the meridional reflections 215 and 143 A during a single twitch of the muscle (Huxley et al., Nature, 1980 284, 140) may be interpreted as a natural consequence of the structural change in the thick filaments. It is concluded therefore that on stimulation of the vertebrate skeletal muscle the thickness filaments undergo a reversible structural change which may reflect the existence of myosin-linked regulation in that type of muscle.


Role of Vascular Smooth Muscle Cell Phenotypic Switching and Calcification in Aortic Aneurysm Formation.
Aortic aneurysm is a vascular disease whereby the ECM (extracellular matrix) of a blood vessel degenerates, leading to dilation and eventually vessel wall rupture. Recently, it was shown that calcification of the vessel wall is involved in both the initiation and progression of aneurysms. Changes in aortic wall structure that lead to aneurysm formation and vascular calcification are actively mediated by vascular smooth muscle cells. Vascular smooth muscle cells in a healthy vessel wall are termed contractile as they maintain vascular tone and remain quiescent. However, in pathological conditions they can dedifferentiate into a synthetic phenotype, whereby they secrete extracellular vesicles, proliferate, and migrate to repair injury. This process is called phenotypic switching and is often the first step in vascular pathology. Additionally, healthy vascular smooth muscle cells synthesize VKDPs (vitamin K-dependent proteins), which are involved in inhibition of vascular calcification. The metabolism of these proteins is known to be disrupted in vascular pathologies. In this review, we summarize the current literature on vascular smooth muscle cell phenotypic switching and vascular calcification in relation to aneurysm. Moreover, we address the role of vitamin K and VKDPs that are involved in vascular calcification and aneurysm. Visual Overview- An online visual overview is available for this article.


The social distancing law project template: a method for jurisdictions to assess understanding of relevant legal authorities.
The Centers for Disease Control and Prevention and the Association of State and Territorial Health Officials selected 17 state and large local jurisdictions on the basis of their proximity to federal quarantine stations and collaborated with their state health department legal counsel to conduct formulaic self-assessments of social distancing legal authorities, create tables of authority, and test and report on the laws' sufficiency (ie, scope and breadth). Select jurisdictions also held tabletop exercises to test public health and law enforcement officials' understanding and implementation of pertinent laws. This report presents findings for Michigan, which completed the legal assessment and tabletop exercise and made several recommendations for change as a result. Officials in Michigan concluded that there are sufficient existing laws to support social distancing measures but that a spectrum of questions remained regarding implementation of these legal authorities. Based on the findings of this assessment, Michigan initiated actions to address areas for improvement. The results of this project highlighted the value of integrally involving the state health department's legal counsel-those most familiar with and who advise on a given state's public health laws-in the periodic identification, assessment, and testing of the state's legal authorities for social distancing and other measures used in response to many public health emergencies.


A new chemical procedure for 32P-labeling of ribonucleic acids at their 5'-ends after isolation.
A new technique, which utilizes the chemical reaction between [32P]diimidazolidate of orthophosphate and the cetyltrimethylammonium salt of high-molecular-weight RNA in nonaqueous dimethyl formamide, has been developed for the 32P-labeling of RNAs after isolation. The radioactive label of high specific activity is introduced onto a phosphorylated 5'-end of the RNA and renders it suitable for 5'-terminal group analysis. When the labeling reaction was applied to the 70S RNA of avian myeloblastosis virus, a labeled 35S RNA was isolated on sucrose-dimethyl sulfoxide gradients without apparent degradation.


Changes in body composition and its relation to muscle strength in 75-year-old men and women: a 5-year prospective follow-up study of the NORA cohort in Göteborg, Sweden.
The purpose of this study was to explore the association between body composition in the elderly and subsequent changes in muscle strength during aging. This was a longitudinal study with a 5-y follow-up. Eighty-seven men (n = 38) and women (n = 49) from a random sample of 75-y-old subjects in the Göteborg part of the Nordic Research on Aging study who were investigated at ages 75 and 80 y and were free from any major diseases at baseline were included. Body composition was estimated from bioelectrical impedance. The maximal isometric strengths of handgrip, arm flexion, and knee extension were measured on the side of the dominant hand while a subject was in a sitting position in an adjustable dynamometer chair. Fat-free mass decreased significantly (P < 0.001) in both sexes, but more in men. Percentage of body fat increased only in men (P < 0.05). Body height decreased in both sexes, but more in women (P < 0.001). Declines in muscle strengths were evident for all muscle groups in both sexes but more prominent in men. It was observed that body composition status at baseline, measured as fat-free mass and fat-free mass index, was a statistically significant predictor for decline in muscle strength, particularly in the extremities. Fat-free mass at age 75 y was associated with lower 5-y decline in muscle strength. This finding underscores the potential importance of fat-free mass for maintaining functional ability during aging.


Engineering electronic properties of graphene by coupling with Si-rich, two-dimensional islands.
Recent theoretical and experimental studies demonstrated that breaking of the sublattice symmetry in graphene produces an energy gap at the former Dirac point. We describe the synthesis of graphene sheets decorated with ultrathin, Si-rich two-dimensional (2D) islands (i.e., Gr:Si sheets), in which the electronic property of graphene is modulated by coupling with the Si-islands. Analyses based on transmission electron microscopy, atomic force microscopy, and electron and optical spectroscopies confirmed that Si-islands with thicknesses of ~2 to 4 nm and a lateral size of several tens of nm were bonded to graphene via van der Waals interactions. Field-effect transistors (FETs) based on Gr:Si sheets exhibited enhanced transconductance and maximum-to-minimum current level compared to bare-graphene FETs, and their magnitudes gradually increased with increasing coverage of Si layers on the graphene. The temperature dependent current-voltage measurements of the Gr:Si sheet showed approximately a 2-fold increase in the resistance by decreasing the temperature from 250 to 10 K, which confirmed the opening of the substantial bandgap (~2.5-3.2 meV) in graphene by coupling with Si islands.


Analysis of insertions and deletions in the gag p6 region of diverse HIV type 1 strains.
Sequence variation in the gag p6 region in subtype B HIV-1 has been associated with changes in viral replication capacity and antiretroviral drug susceptibility. We examined sequence variation in the HIV-1 gag p6 region using plasma samples from 22 individuals with non-subtype B HIV-1 infection [subtypes A, C, D, F, and G, and circulating recombinant forms (CRFs) CRF01-AE and CRF02_AG]. An additional 105 gag sequences from the Los Alamos National Laboratory database were also analyzed. Extensive length variation was observed in the p6 gag region. Specific patterns of insertions and deletions were observed in different subtypes and CRFs, and no two subtypes or CRFs had the same general pattern. PTAP duplications were more common in subtype C than other strains (3 of 14 in subtype C vs. 2 of 113 in other strains, p = 0.004), and KQE duplications were seen only in subtype B. Further studies are needed to determine whether such genotypic differences influence viral replication capacity, antiretroviral drug susceptibility, or other phenotypic properties of these strains.


Optimal erythropoietin expression in human hepatoma cell lines requires activation of multiple signalling pathways.
Hypoxia is thought to be a common precursor of coronary artery disease and malignant tumors, both diseases representing the leading causes of death in industrial nations. So far, investigations of oxygen-regulated erythropoietin (EPO) gene expression in the human hepatoma cell lines Hep3B and HepG2 allowed many important insights into the mechanisms of oxygen-sensing, signalling and regulation of an increasing number of oxygen-responsive genes. To differentiate the various signalling pathways involved in EPO production by these two cell lines, we examined several factors that positively influenced EPO expression. The results demonstrate a keen differential effect of cell density and oxygen concentration on EPO induction in Hep3B compared to HepG2 cells. Using optimized cell culture conditions, EPO production rates as high as 1 U EPO per 10(6) Hep3B cells in 24 h could be achieved. We also found a moderate but reproducible positive effect of CoCl2 on hypoxia-induced EPO expression in Hep3B but a negative CoCl2 effect on hypoxic induction in HepG2 cells. CoCl2 inhibited cell growth in a concentration-dependent manner. Interleukin-6 was synergistic with hypoxia on EPO induction in Hep3B as well as HepG2 cells, and dexamethasone enhanced this effect in Hep3B but not in HepG2 cells. The moderate CoCl2-dependent increase of EPO production observed in hypoxic Hep3B cells might indicate that CoCl2 and hypoxia do not necessarily act via, identical signalling pathways.


Comparison of simulated cone beam computed tomography to conventional helical computed tomography for imaging of rhinosinusitis.
Cone beam computed tomography (CBCT) has emerged as a low radiation dose alternative to traditional computed tomography (CT) to evaluate the paranasal sinuses. The purpose of our study was to determine how often clinically important findings would be missed if CBCT was used routinely for sinus imaging. Retrospective review. We evaluated all maxillofacial CT scans performed for sinusitis over a 1-year period. The original multidetector CT (MDCT) images were reviewed retrospectively. A theoretical CBCT was then created from the original study utilizing only the bone algorithm images and separately reviewed. We calculated the proportion of abnormal findings that were identified on the MDCT but would have been missed by the theoretical CBCT, and reviewed the medical record to determine which potentially missed findings would have changed management. Radiation dose from the MDCT scanners was calculated and compared to published dose estimates for the paranasal sinuses on CBCT. Maxillofacial CTs from 361 consecutive patients were included, of which 12 (3.3%) demonstrated findings that would have been missed on the theoretical CBCT. Of those, four (1.1%) would have resulted in a change in management. Effective radiation dose for our scanners ranged from 0.67 mSv to 2.15 mSv, compared to a published estimated dose of 0.2 mSV for CBCT. In the majority of patients undergoing simulated CBCT for evaluation of sinusitis, incidental findings of soft-tissue disease were rare. With appropriate selection of patients, CBCT can offer substantial radiation dose reduction and may provide a viable alternative to standard MDCT sinus imaging protocols.


Antiproliferative effect of 1,4-phenylenebis(methylene)selenocyanate (p-XSC) on colonic epithelium of patients with adenomatous polyps in vitro.
We have consistently shown that the organoselenium compound 1,4-phenylenebis(methylene)selenocyanate (p-XSC) is a superior cancer chemopreventive agent and less toxic than selenite or certain naturally-occurring selenoamino acids. To elucidate the effects of p-XSC on human colonic mucosa, biopsies from endoscopically normal sigmoid colon of 30 patients with adenomatous polyps were incubated with p-XSC at concentrations of 1, 2 and 5 micromol/l dissolved in dimethylsulphoxide (DMSO). Biopsies incubated with DMSO or pure culture medium served as a control. Proliferating cells were labelled by bromodeoxyuridine immunohistochemistry and the labelling index (LI) was computed. Upper crypt labelling index (LI of crypt compartments 4+5) and Phih value, which are both discriminators of the expansion of the proliferative zone, were significantly lower after incubation with 1 and 5 micromol/l p-XSC, respectively (LI 4+5: 0.8 and 1.0; Phih value: 2.1 and 2.4), as compared with DMSO (LI 4+5: 3.6 and 4.5; Phih value: 7.0 and 8.3) or culture medium (LI 4+5: 3.3 and 4.5; Phih value: 7.2 and 8.1) (P<0.005 and P<0.05 by Friedman's block test). A trend towards lower levels of LI 4+5 (P=0.059) and Phih value (P=0.075) were seen after 2 micromol/l p-XSC incubation compared with DMSO. Since hyperproliferation of colonic crypt cells with expansion of the proliferative zone is regarded as a biomarker of increased cancer risk, the antiproliferative effects of p-XSC especially on upper crypt LI and Phih value may indicate a possible protective effect of this organoselenium compound in the prevention of human colon cancer development.


Evaluation of surgical performance during laparoscopic incisional hernia repair: a multicenter study.
Laparoscopic incisional hernia repair (LIHR) is a common procedure requiring advanced laparoscopic skills. This study aimed to develop a procedure-specific tool to assess the performance of LIHR and to evaluate its reliability and validity. The Global Operative Assessment of Laparoscopic Skills-Incisional Hernia (GOALS-IH) is a 7-item global rating scale developed by experts to evaluate the steps of LIHR (placement of trocars, adhesiolysis, estimation of mesh size and shape, mesh orientation and positioning, mesh fixation, knowledge and autonomy in use of instruments, overall competence), each rated on a 5-point Likert scale. During LIHR, 13 attending surgeons and fellows experienced in minimally invasive surgery (MIS) and 19 novice surgeons (postgraduate years [PGYs], 3-5) were evaluated at four teaching hospitals by the attending surgeon, a trained observer, and self-assessment using GOALS-IH, and by a previously validated 5-item general laparoscopic rating scale (GOALS). Interrater reliability was assessed by intraclass correlation (ICC), and internal consistency of rating items was assessed by Cronbach's alpha. Known-groups construct validity was assessed by using the t-test and by correlating of the number of self-reported LIHR cases with the total score. Concurrent validity was assessed by correlating the GOALS-IH score with the GOALS general rating scale. Data are presented as mean and 95% confidence interval (CI). Interrater reliability for the total GOALS-IH score was 0.79 (95% CI, 0.60-0.89) between observers and attending surgeons, 0.81 (95% CI, 0.58-0.92) between participants and attending surgeons, and 0.89 (95% CI, 0.76-0.96) between participants and observers. Internal consistency was high (Cronbach's alpha, 0.93). Experienced surgeons performed significantly better than novices as assessed by GOALS-IH (31; 95% CI, 29-33 vs. 21; 95% CI, 19-24; p < 0.01). Very good correlation was found between GOALS-IH and previous LIHR experience (r = 0.82; p < 0.01) and strong correlation between GOALS-IH and generic GOALS total scores (r = 0.90; p < 0.01). Surgical performance during clinical LIHR can be assessed reliably using GOALS-IH. Results can be used to provide formative feedback to the surgeon and to identify steps of the operation that would benefit from specific educational interventions.


[Pharmacological characterization and mechanisms of the novel antidepressive- and/or anxiolytic-like substances identified from Perillae Herba].
The leaves of Perilla frutescens Britton var. acuta Kudo (Perillae Herba) are commonly found in traditional oriental herbal medicines, which are primarily used to treat depression and anxiety-related disorders. We have recently identified rosmarinic acid as well as its major metabolite caffeic acid as novel antidepressive and/or anxiolytic-like substances from components within Perillae Herba. The present article will review our previous findings and also provide a view of future studies. Behavioral as well as analytical studies have demonstrated that first, the extracts from Perillae Herba, which produced an antidepressive-like effect in the forced swimming test, contained abundant rosmarinic acid; second, the extracts from another species of Perillae Herba, which contains only low levels of rosmarinic acid, did not have an antidepressive-activity; third, rosmarinic acid and caffeic acid themselves also produced an antidepressive-like effect; finally, both compounds showed an anxiolytic-like activity in the conditioned fear stress test. Neurochemical studies have also revealed that neither rosmarinic acid nor caffeic acid affects the uptake of monoamines or monoamine oxidase activity, which may underlie the therapeutic value of existing clinically effective antidepressants. Furthermore, it has been found in the recent study that caffeic acid produces antidepressive- and/or anxiolytic-like effects through the modulation of the alpha1A-adrenoceptor-mediated signal transductions and also attenuates the down-regulation of BDNF transcription that results from the exposure to forced swimming stress. These results suggest that rosmarinic acid and caffeic acid may produce antidepressive- and/or anxiolytic-like effects via some mechanism(s) other than those of drugs that now clinically used. Therefore, further studies on the mechanisms involved in the antidepressive- and/or anxiolytic-like properties of both substances could help to explain the pathophysiology underlying depression and anxiety disorders, and pave the way for the development of new therapeutic drugs.


Aplastic anemia and pure red cell aplasia.
The role of known hematopoietic growth factors in the pathogenesis of aplastic anemia and congenital hypoplastic anemia has been extensively studied and no evidence has been obtained that deficiency of these factors contributes to the hypoproliferative state in these disorders. Clonal hematopoiesis seems to be present at least in a small percentage of cases of aplastic anemia, a finding that needs further investigation. Androgens were shown to be beneficial only for women with aplastic anemia treated with antilymphocyte globulin. Unrelated-donor bone marrow transplantation is becoming a realistic approach for children and very young adults with aplastic anemia, but in older groups the survival is very poor. New observations on abnormalities of lymphokines and cytokines in Fanconi's anemia have been described, but their pathogenetic significance remains unknown. A large number of studies have excluded the possibility that abnormalities of c-kit/SCF genes and their expression are responsible for the erythroid aplasia in Diamond-Blackfan syndrome. Cyclosporine was found to be an effective treatment for pure red cell aplasia associated with chronic lymphocytic leukemia. The cell membrane receptor for B19 parvovirus has been identified as the P antigen. Long-term studies showed that in 20% of patients with homozygous sickle cell disease, infection by B19 does not cause erythroid aplasia.


Damage Evolution in Complex-Phase and Dual-Phase Steels during Edge Stretching.
The role of microstructural damage in controlling the edge stretchability of Complex-Phase (CP) and Dual-Phase (DP) steels was evaluated using hole tension experiments. The experiments considered a tensile specimen with a hole at the center of specimen that is either sheared (sheared edge condition) or drilled and then reamed (reamed edge condition). The damage mechanism and accumulation in the CP and DP steels were systematically characterized by interrupting the hole tension tests at different strain levels using scanning electron microscope (SEM) analysis and optical microscopy. Martensite cracking and decohesion of ferrite-martensite interfaces are the dominant nucleation mechanisms in the DP780. The primary source of void nucleation in the CP800 is nucleation at TiN particles, with secondary void formation at martensite/bainite interfaces near the failure strain. The rate of damage evolution is considerably higher for the sheared edge in contrast with the reamed edge since the shearing process alters the microstructure in the shear affected zone (SAZ) by introducing work-hardening and initial damage behind the sheared edge. The CP microstructures were shown to be less prone to shear-induced damage than the DP materials resulting in much higher sheared edge formability. Microstructural damage in the CP and DP steels was characterized to understand the interaction between microstructure, damage evolution and edge formability during edge stretching. An analytical model for void evolution and coalescence was developed and applied to predict the damage rate in these rather diverse microstructures.


Notch1 and Numb genes are inversely expressed as oligodendrocytes differentiate.
The Notch1 pathway plays a fundamental role during the establishment of cell fates in the central nervous system (CNS) by regulating neural cell differentiation. In oligodendrocytes (OLs), Notch1 activity prevents these cells from becoming terminally mature, thereby influencing CNS myelination. Little is known of how OLs regulate the expression of this receptor at the gene level or if OLs have mechanisms to control the level of intracellular activity of the Notch1 pathway. In this study, we have found that Notch1 gene expression was higher in proliferative OL progenitor cells (OPCs) and was reduced when cells were forced to withdraw from the cell cycle and became mature, indicating that Notch1 gene expression is developmentally regulated in OLs. We observed that the blockade of terminal differentiation of OPCs by incubation with Delta1, an activator of Notch1, was a dominant process and OL-differentiating signals such as thyroid hormone could not overcome this inhibition in culture. This suggests that a downregulation of the Notch1 pathway might be required to allow OPCs to enter terminal differentiation. We also provide evidence that OPCs and OLs express the Numb gene, a known negative regulator of Notch1 activity. In vivo, Numb was found in postnatal OLs from cerebellar and cerebral white matter. In vitro, Numb expression showed to be inversely correlated to that of Notch1, with higher levels of Numb proteins in mature OLs, in association with myelin-like membranes.


Updates in Cardiac Arrest Resuscitation.
There are approximately 350,000 out-of-hospital cardiac arrests and 200,000 in-hospital cardiac arrests annually in the United States, with survival rates of approximately 5% to 10% and 24%, respectively. The critical factors that have an impact on cardiac arrest survival include prompt recognition and activation of prehospital care, early cardiopulmonary resuscitation, and rapid defibrillation. Advanced life support protocols are continually refined to optimize intracardiac arrest management and improve survival with favorable neurologic outcome. This article focuses on current treatment recommendations for adult nontraumatic cardiac arrest, with emphasis on the latest evidence and controversies regarding intracardiac arrest management.


Polarization-insensitive ultralow-power second-harmonic generation frequency-resolved optical gating.
We demonstrate polarization-insensitive ultralow-power second-harmonic generation frequency-resolved optical gating (FROG) measurements with a fiber-pigtailed, aperiodically poled lithium niobate waveguide. By scrambling the polarization much faster than the measurement integration time, we eliminate the impairment that frequency-independent random polarization fluctuations induce in FROG measurements. As a result we are able to retrieve intensity and phase profiles of few hundred femtosecond optical pulses with 50 MHz repetition rates at 5.2 nW coupled average power without control of the input polarization.


Genetics of hybrid inviability and sterility in Drosophila: dissection of introgression of D. simulans genes in D. melanogaster genome.
Interspecific crosses between Drosophila melanogaster and Drosophila simulans usually produce sterile unisexual hybrids. The barrier preventing genetic analysis of hybrid inviability and sterility has been taken away by the discovery of a D. simulans strain which produces fertile female hybrids. D. simulans genes in the cytological locations of 21A1 to 22C1-23B1 and 30F3-31C5 to 36A2-7 have been introgressed into the D. melanogaster genetic background by consecutive backcrosses. Flies heterozygous for the introgression are fertile, while homozygotes are sterile both in females and males. The genes responsible for the sterility have been mapped in the introgression. The male sterility is caused by the synergistic effect of multiple genes, while the female sterility genes have been localized to a 170 kb region (32D2 to 32E4) containing 20 open reading frames. Thus, the female sterility might be attributed to a single gene with a large effect. We have also found that the Lethal hybrid rescue mutation which prevents the inviability of male hybrids from the cross of D. melanogaster females and D. simulans males cannot rescue those carrying the introgression, suggesting that D. simulans genes maybe non-functional in this hybrid genotype. The genes responsible for the inviability have not been separated from the female sterility genes by recombination.


MerTK expressing hepatic macrophages promote the resolution of inflammation in acute liver failure.
Acute liver failure (ALF) is characterised by overwhelming hepatocyte death and liver inflammation with massive infiltration of myeloid cells in necrotic areas. The mechanisms underlying resolution of acute hepatic inflammation are largely unknown. Here, we aimed to investigate the impact of Mer tyrosine kinase (MerTK) during ALF and also examine how the microenvironmental mediator, secretory leucocyte protease inhibitor (SLPI), governs this response. Flow cytometry, immunohistochemistry, confocal imaging and gene expression analyses determined the phenotype, functional/transcriptomic profile and tissue topography of MerTK+ monocytes/macrophages in ALF, healthy and disease controls. The temporal evolution of macrophage MerTK expression and its impact on resolution was examined in APAP-induced acute liver injury using wild-type (WT) and Mer-deficient (Mer-/-) mice. SLPI effects on hepatic myeloid cells were determined in vitro and in vivo using APAP-treated WT mice. We demonstrate a significant expansion of resolution-like MerTK+HLA-DRhigh cells in circulatory and tissue compartments of patients with ALF. Compared with WT mice which show an increase of MerTK+MHCIIhigh macrophages during the resolution phase in ALF, APAP-treated Mer-/- mice exhibit persistent liver injury and inflammation, characterised by a decreased proportion of resident Kupffer cells and increased number of neutrophils. Both in vitro and in APAP-treated mice, SLPI reprogrammes myeloid cells towards resolution responses through induction of a MerTK+HLA-DRhigh phenotype which promotes neutrophil apoptosis and their subsequent clearance. We identify a hepatoprotective, MerTK+, macrophage phenotype that evolves during the resolution phase following ALF and represents a novel immunotherapeutic target to promote resolution responses following acute liver injury.


How are medication related problems managed in primary care? An exploratory study in patients with diabetes and primary care providers.
Medication self-management is important for patients who are controlling diabetes. Achieving medication self-management goals, may depend on treatment complexity and patients' capacities such as health literacy, knowledge and attitude. The aims of this study were to explore how patients with diabetes self-manage their medications, how patients seek support when experiencing problems and how primary healthcare providers identify patients' medication related problems and provide support. Semi-structured interviews were conducted among patients with diabetes receiving primary care and with their primary healthcare providers - GPs, nurses, pharmacists and technicians - between January and June 2017. A purposive sampling strategy was used to identify and select participants. An interview guide based on the Cycle of Complexity model was developed. Interviews were audiotaped and transcribed verbatim. Transcripts were coded with a combination of deductive and inductive codes. A thematic analysis was performed to identify categories and themes in the data. Findings were compared with the Cycle of Complexity model. Twelve patients and 27 healthcare providers were included in the study. From the transcripts 95 codes, 6 categories and 2 major themes were extracted. Patients used practical solutions and gaining knowledge to manage their medication. Their problems were often related to stress and concerns about using medications. A trusted relationship with the healthcare provider was essential for patients to share problems and ask for support. Informal support was sought from family and peer-patients. Healthcare providers perceive problem identification as challenging. They relied on patients coming forward, computer notifications, clinical parameters and gut-feeling. Healthcare providers were able to offer appropriate support if a medication management problem was known. Patients are confident of finding their way to manage their medications. However, sharing problems with healthcare providers requires a trusted relationship. This is acknowledged by both patients and healthcare providers.


[Health-related quality of life before and one year after operation for morbid obesity].
Several studies have shown that morbid obese patients have poor health-related quality of life. The aim of this study was to investigate the effect of bariatric surgery (biliopancreatic diversion with duodenal switch) on health-related quality of life for morbidly obese patients. 48 patients (26 women and 22 men, mean age 37.4, mean BMI 51.5) completed the Short Form 36 (SF-36) before and one year after surgery. Paired t-tests were used to analyze differences in SF-36 scores before and one year after surgery. Morbidly obese patients had very poor health-related quality of life (SF-36) before surgery compared with norms (p < 0.001). One year after surgery, all SF-36 dimensions had improved significantly (p < 0.001). The morbidly obese patients in this study had very poor health-related quality of life compared to norms, but they experienced considerable improvement after bariatric surgery.


[Problems in primary and secondary selection of personnel for scientific research].
The author presents his own observations and reflections about the problems which appear with primary and secondary selection and election, in other words, with the selection, election and reelection of candidates, meaning personnel for scientific-research work at institutes and clinics of medical faculties in our country. He is in favor of the introduction of unique, agreed on, scientific and professional criteria, and with that, their strict obeying, without the influence of para-scientific factors, with the elimination of the attempts of such negative occurrences and attempted activities. In the process of secondary selection (reelection) the author recommends the strict obedience of principles of expertly proven scientific and professional qualities of the candidate, with giving priority and propulsive direction to the young, contemporarily educated and efficient workers in scientific research, and the principled elimination from institutes and clinics of such people which have not, up to now, actually proven themselves in the scientific and professional field. With this, broad spaces open for the constant selection of new young personnel and the removal of the insufficiently qualified "plethora" which blocks the further development and efficient work of scientific-research institutions at medical faculties.


Functional MRI evidence for language plasticity in adult epileptic patients: Preliminary results.
The present fMRI study explores the cerebral reorganisation of language in patients with temporal lobe epilepsy, according to the age of seizures onset (early or late) and the hippocampal sclerosis (associated or not). Seven right-handed control volunteers and seven preoperative adult epileptic patients performed a rhyme decision (language condition) and a visual detection (control condition) tasks in visually presented words and unreadable characters, respectively. All patients were left hemisphere dominant for language. Appropriate statistical analyses provided the following preliminary results: (1) patients compared with healthy subjects showed lower degree of hemispheric lateralization with supplementary involvement of the right hemisphere; (2) the degree of hemispheric specialization depends on the considered region; (3) patients with early seizures show signs of temporal and parietal reorganization more frequently than patients with late onset of seizures; (4) patients with early seizures show a tendency for intra-hemispheric frontal reorganisation; (5) associated hippocampal sclerosis facilitates the inter-hemispheric shift of temporal activation. Although our patients were left hemisphere predominant for language, the statistical analyses indicated that the degree of lateralization was significantly lower than in healthy subjects. This result has been considered as the indication of atypical lateralization of language.


Acculturation and perceived stress in HIV+ immigrants: depression symptomatology in Asian and Pacific Islanders.
Asians and Pacific Islanders (API) are among the fastest growing minority groups within the USA, and this growth has been accompanied by an increase in HIV incidence. Between 2000 and 2010, the API HIV infection rate increased from 4.5% to 8.7%; however, there is a paucity of HIV-related research for this group, and even less is known about the prevalence and correlates of antiretroviral therapy adherence behavior, quality of life, impact of stress, and efficacious self-management among HIV+ API Americans. This paper examines how acculturation and perceived stress affect depression symptomatology and treatment seeking in the HIV+ API population. A series of cross-sectional audio computer-assisted self-interviews were conducted with a convenience sample of 50 HIV+ API (29 in San Francisco and 21 in New York City). The relationship between acculturation and perceived stress was analyzed, and the results indicate that for those HIV+ API who reported low or moderate acculturation (as compared to those who reported high acculturation), stress was significantly mediated by depression symptomology. Interventions to address acculturation and reduce perceived stress among API generally and Asians specifically are therefore needed.


Effects of dietary functional ingredients and packaging methods on sensory characteristics and consumer acceptance of irradiated turkey breast meat.
Raw and cooked breast patties from turkeys fed 8 different diets [control; 200 IU/kg of vitamin E (VE); 0.3 mg/kg of Se; 2.5% conjugated linoleic acids (CLA); 200 IU/kg of VE + 0.3 mg/kg of Se; 200 IU/kg of VE + 2.5% CLA; 0.3 mg/kg of Se + 2.5% CLA; and 200 IU/kg of VE + 0.3 mg/kg of Se + 2.5% CLA] were treated with 2 irradiation doses (0 and 1.5 kGy) and 2 packaging methods (vacuum and aerobic). Raw and cooked samples from 32 treatments were tested by 8 trained sensory panelists for turkey aroma and irradiation off-aroma. Based on the sensory scores, the 3 dietary treatments producing the most and the least off-aroma were selected and used for a consumer acceptance study. Sensory results of raw meat showed that turkey aroma was intense in aerobically packaged meat, whereas irradiation off-aroma was intense with vacuum packaging. Raw meats from dietary treatments containing CLA (CLA, VE + CLA, Se + CLA, VE + Se + CLA) had greater turkey aroma scores, whereas those containing VE (VE and VE + Se) had lower scores than the control. Dietary treatments containing VE (VE, VE + Se, VE + Se + CLA) significantly lowered (P < 0.05) irradiation off-aroma in raw turkey breast meat, whereas CLA increased it, especially when the meats were packaged aerobically. In cooked meat, however, irradiation and packaging had no effect on turkey meat aroma and irradiation off-aroma. Cooked meat from turkeys supplemented with VE (VE and VE + Se) had less (P < 0.05) irradiation off-odor than other dietary treatments. Dietary CLA increased the irradiation off-aroma in cooked meat, which could not be reduced, even when VE and Se were combined in the diet. Irradiation off-aroma of raw meat was not pleasant for most consumers, and dietary supplementation of VE and VE + Se improved consumer acceptance of irradiated raw meat. For cooked meat samples, consumers preferred both color and flavor of irradiated meat to nonirradiated meat.


The molecular basis of carcinogenesis: understanding the cell cycle clock.
The cell cycle clock is the central controller of cell proliferation that governs the progress of the cell through its growth cycle, its exit from the active cycle, and its decision to differentiate. Components of the clock are found to be functioning in an aberrant fashion in many types of malignancies. Notable among these is the retinoblastoma protein, pRB, which acts to restrain proliferation in normal cells and suffers inactivation in many types of tumour cells. Its activity is controlled by D-type cyclins in various cell types. We have deleted one of these cyclins--cyclin D1--from the mouse germline and find that its absence leads to a limited range of defects including hypoplastic retinae and the inability of the mammary epithelium to respond to pregnancy-associated hormonal stimulation. Cyclin D1 is overexpressed in many human breast cancers, pointing to a highly specific association of this cell cycle clock component with mammary cell proliferation.


Distribution and morphology of the catecholaminergic neural elements in the human hypothalamus.
Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreactive (IR) perikarya and fibers are widely distributed in the human hypothalamus. Since TH is the key and rate-limiting enzyme for catecholaminergic synthesis, these IR neurons may represent dopaminergic, noradrenergic or adrenergic neural elements. However, the distribution and morphology of these neurotransmitter systems in the human hypothalamus is not entirely known. Since the different catecholaminergic systems can be detected by identifying the neurons containing the specific key enzymes of catecholaminergic synthesis, in the present study we mapped the catecholaminergic elements in the human hypothalamus using immunohistochemistry against the catecholaminergic enzymes, TH, dopamine beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT). Only a few, PNMT-IR, adrenergic neuronal elements were found mainly in the infundibulum and the periventricular zone. DBH-IR structures were more widely distributed in the human hypothalamus occupying chiefly the infundibulum/infundibular nucleus, periventricular area, supraoptic and paraventricular nuclei. Dopaminergic elements were detected by utilizing double label immunohistochemistry. First, the DBH-IR elements were visualized; then the TH-IR structures, that lack DBH, were detected with a different chromogen. In our study, we conclude that all of the catecholaminergic perikarya and the majority of the catecholaminergic fibers represent dopaminergic neurons in the human hypothalamus. Due to the extremely small number of PNMT-IR, adrenergic structures in the human hypothalamus, the DBH-IR fibers represent almost exclusively noradrenergic neuronal processes. These findings suggest that the juxtapositions between the TH-IR and numerous peptidergic systems revealed by previous reports indicate mostly dopaminergic synapses.


Lung capillary changes in hepatic cirrhosis in rats.
The hypoxemia of the hepatopulmonary syndrome may result from dilated intrapulmonary vascular segments. Knowledge of the size, density, and branching frequency of the lung capillaries might confirm this hypothesis and suggest that the pathogenesis may involve vascular dilatation or angiogenesis. To investigate these changes, the common bile duct of rats was tied off to cause biliary cirrhosis. Later (4 wk), the pulmonary vasculature of these animals was cast, and the casts were studied with scanning electron microscopy. In the ligated animals, evidence for enhancement of the bronchial to pulmonary circulation was found: cast vasa vasorum of the pulmonary arteries and cast bronchial veins emptying into pulmonary veins. Ligated animals had more adherent intracapillary cells per alveolus than the sham-operated animals. The diameters of all capillary beds were larger in the ligated animals. The alveolar capillary density was increased, but the branching frequency was not. A few areas suggesting angiogenesis were found. Induction of biliary cirrhosis enhances the pulmonary-systemic circulation, increases intracapillary adherent cells, capillary diameter, and density, and may be associated with angiogenesis.


Sexual Dysfunction and Incidence of Depression in Multiple Sclerosis Patients
Multiple sclerosis (MS) is one of the most common diseases of the central nervous system and usually occurs at the age when people would be expected to be in the prime of their sexual lives. In everyday practice, sexual dysfunction is underestimated because clinicians mostly concentrate on the classic neurologic deficits and often overlook symptoms that can seriously affect the quality of life. Our study included 98 patients (42 men and 56 women, mean age 35±12 years) with relapse from our MS register, with established diagnosis of relapsing remitting multiple sclerosis according to McDonald criteria. Patients completed the questionnaires (Sexual Satisfaction Scale, SSS and Beck Depression Scale BDS), and underwent neurological assessment (Expanded Disability Status Scale, EDSS). All patients were in the group with EDSS 2 to 4 points (mobile patients). There was no statistically significant difference in BDS and SSS values according to EDSS score. Correlation coefficients were calculated (BDS and SSS) for men (p=0.42) and women (p=0.44), yielding positive correlation. There was no statistically significant difference in BDS and SSS values according to gender, disease duration or immunomodulatory therapy. In our group of patients, despite low EDSS score (fully ambulatory without aid, self sufficient patients) we found positive correlation between sexual dysfunction and depression, showing that even in such patients the quality of life can be decreased. In conclusion, sexual dysfunction and depression are mostly under-recognized by neurologists because they are not part of routine testing; therefore, some additional questionnaires should be used in the evaluation in MS patients, even those with low EDSS score, in order to improve their quality of life.


Prevalence of nutritional wasting in populations: building explanatory models using secondary data.
To understand how social context affects the nutritional status of populations, as reflected by the prevalence of wasting in children under 5 years of age from Africa, Latin America, and Asia; to present a systematic way of building models for wasting prevalence, using a conceptual framework for the determinants of malnutrition; and to examine the feasibility of using readily available data collected over time to build models of wasting prevalence in populations. Associations between prevalence of wasting and environmental variables were examined in the three regions. General linear mixed models were fitted using anthropometric survey data for countries within each region. Low birth weight (LBW), measles incidence, and access to a safe water supply explained 64% of wasting variability in Asia. In Latin America, LBW and survey year explained 38%; in Africa, LBW, survey year, and adult literacy explained 7%. LBW emerged as a predictor of wasting prevalence in all three regions. Actions regarding women's rights may have an effect on the nutritional status of children since LBW seems to reflect several aspects of the conditions of women in society. Databases have to be made compatible with each other to facilitate integrated analysis for nutritional research and policy decision-making. In addition, the validity of the variables representing the conceptual framework should be improved.


Minimally Invasive Unilateral Laminectomy for Bilateral Decompression.
Unilateral laminectomy for bilateral decompression (ULBD) is a recently popularized minimally invasive surgical technique for decompression of the spinal canal. With the patient prone on the spinal table of your choice, use an image intensifier to determine the incision position and then position the retractor of your choice to identify the inferior aspect of the superior lamina. Begin the laminotomy on the approach side, drilling to identify the ligamentum flavum on the approach side, and remove bone up to the superior attachment of the ligamentum flavum. To gain access to the contralateral side of the canal for bilateral decompression, remove enough of the spinous process to gain access to the midline and contralateral ligamentum flavum. The superior aspect of the decompression usually corresponds with the superior ligamentum flavum attachment, except in certain cases such as when a facet joint cyst extends beyond the limits of the ligamentum flavum; removal of the upper limit of the ligamentum flavum provides an important landmark to confirm the superior limit of the decompression. Detach the ligamentum flavum from the facet joint on the approach side using a combination of angled curets and Kerrison rongeurs; a partial medial facetectomy, or removal of adequate facet hypertrophy, on the approach side is necessary to expose the traversing nerve root. Decompression of the thecal sac on the contralateral side of the canal is the potentially dangerous aspect of the procedure, with the highest risk of dural injury and a cerebrospinal fluid leak; thus, create enough room on the ipsilateral side so that instruments can be safely introduced into the canal for the contralateral decompression. Reducing the paraspinal muscle dissection substantially reduces iatrogenic muscle injury and blood loss, and oozing from the bone removal can be easily controlled with bone wax or a variety of hemostatic agents. Closure of a unilateral muscle exposure is rapid and the use of wound drainage is very rare, further reducing operative time as well as exposure to complications related to wound drains and subsequent infection risk. One of us (R.M.) and colleagues5 conducted a prospective randomized trial comparing ULBD with open laminectomy for degenerative lumbar spinal stenosis in 54 patients (27 in each arm of the study) treated from 2007 to 2009.


Effect of ablation or inhibition of stromal matrix metalloproteinase-9 on lung metastasis in a breast cancer model is dependent on genetic background.
Matrix metalloproteinases (MMP) are a family of enzymes with a myriad of functions. Lately, we have come to realize that broad-spectrum inhibition of these enzymes, as was tried unsuccessfully in multiple phase III trials in cancer patients, is likely unwise given the protumorigenic and antitumorigenic functions of various family members. Here, we used the multistage mammary tumor model MMTV-PyVT to investigate roles for either MMP7 or MMP9 in tumor progression. We found no effect of genetic ablation of MMP7 or MMP9 on the multifocal tumors that developed in the mammary glands. Lack of MMP7 also had no effect on the development of lung metastases, suggesting that MMP7 is irrelevant in this model. In contrast, MMP9 deficiency was associated with an 80% decrease in lung tumor burden. The predominant cellular source of MMP9 was myeloid cells, with neutrophils being the largest contributor in tumor-bearing lungs. Experimental metastasis assays corroborated the role of host-derived MMP9 in lung metastasis and also facilitated determination of a time frame most relevant for the MMP9-mediated effect. The lung tumors from MMP9-deficient mice showed decreased angiogenesis. Surprisingly, the antimetastatic outcome of MMP9 ablation seemed to be dependent on strain. Only mice that had genetic background derived from C57BL/6 showed reduced metastasis, whereas mice fully of the FVB/N background showed no significant effect. These strain-specific responses were also observed in a study using a highly selective pharmacologic inhibitor of MMP9 and thus suggest that responses to MMP inhibition are controlled by genetic differences.


Sensitivity of fecal occult blood testing in the cat.
Objectives The impact of dietary factors on fecal occult blood (FOB) testing has been previously evaluated in cats, but the analytical sensitivity of this point-of-care test remains unexamined. The primary goal of this study was to assess the analytical sensitivity of the FOB test in cats. Methods Five cats were used in a repeated measures study. Following oral administration of blood, feces were collected and tested every 12 h for FOB and melena. All cats were fed an animal protein-free diet starting the week before entry into the study. Blood was administered on a milligram of hemoglobin per kilogram of body weight basis, and dosed at 1.5, 3, 15, 30 and 45 mg/kg hemoglobin in series with a wash-out period between each trial. Results FOB was detected in one cat at 1.5 mg/kg hemoglobin, three cats at 3 mg/kg hemoglobin and in all five cats at 15, 30 and 45 mg/kg hemoglobin. Melena was noted in one cat at 30 mg/kg and four cats at 45 mg/kg, but not at lower doses. Conclusions and relevance Administration of 15 mg/kg hemoglobin (equivalent to about 1.5 ml blood) was sufficient for positive results in all cats. However, detection occurred with as little as 1.5 mg/kg hemoglobin. Thus, FOB has good analytical sensitivity in cats under appropriate clinical situations.


Genetic diversity and selection in Puerto Rican horses.
Since the first Spanish settlers brought horses to America centuries ago, several local varieties and breeds have been established in the New World. These were generally a consequence of the admixture of the different breeds arriving from Europe. In some instances, local horses have been selectively bred for specific traits, such as appearance, endurance, strength, and gait. We looked at the genetics of two breeds, the Puerto Rican Non-Purebred (PRNPB) (also known as the "Criollo") horses and the Puerto Rican Paso Fino (PRPF), from the Caribbean Island of Puerto Rico. While it is reasonable to assume that there was a historic connection between the two, the genetic link between them has never been established. In our study, we started by looking at the genetic ancestry and diversity of current Puerto Rican horse populations using a 668 bp fragment of the mitochondrial DNA D-loop (HVR1) in 200 horses from 27 locations on the island. We then genotyped all 200 horses in our sample for the "gait-keeper" DMRT3 mutant allele previously associated with the paso gait especially cherished in this island breed. We also genotyped a subset of 24 samples with the Illumina Neogen Equine Community genome-wide array (65,000 SNPs). This data was further combined with the publicly available PRPF genomes from other studies. Our analysis show an undeniable genetic connection between the two varieties in Puerto Rico, consistent with the hypothesis that PRNPB horses represent the descendants of the original genetic pool, a mix of horses imported from the Iberian Peninsula and elsewhere in Europe. Some of the original founders of PRNRB population must have carried the "gait-keeper" DMRT3 allele upon arrival to the island. From this admixture, the desired traits were selected by the local people over the span of centuries. We propose that the frequency of the mutant "gait-keeper" allele originally increased in the local horses due to the selection for the smooth ride and other characters, long before the PRPF breed was established. To support this hypothesis, we demonstrate that PRNPB horses, and not the purebred PRPF, carry a signature of selection in the genomic region containing the DMRT3 locus to this day. The lack of the detectable signature of selection associated with the DMRT3 in the PRPF would be expected if this native breed was originally derived from the genetic pool of PRNPB horses established earlier and most of the founders already had the mutant allele. Consequently, selection specific to PRPF later focused on allels in other genes (including CHRM5, CYP2E1, MYH7, SRSF1, PAM, PRN and others) that have not been previously associated with the prized paso gait phenotype in Puerto Rico or anywhere else.


Identification of TRDV-TRAJ V domains in human and mouse T-cell receptor repertoires.
Here, we describe the identification of two T-cell receptors (TRs) containing TRDV genes in their TRA chains, the first one in human and the second one in mouse. First, using 5'RACE on a mixed lymphocyte-tumor cell culture (MLTC), we identified TRDV1 5'-untranslated region (UTR) and complete coding sequence rearranged productively to TRAJ24. Single-cell TR RNA sequencing (RNA-seq) of the MLTC, conducted to identify additional clonotypes, revealed that the analysis software detected the hybrid TRDV-TRAJ TRA (TRA) chain but excluded it from the final results. In a separate project, we performed TR sequencing of tumor-infiltrating lymphocytes (TILs) in a murine tumor model. Here, the predominant clonotype contained a TRA chain with a TRDV2-2-TRAJ49 rearrangement. Again, the hybrid TRA chain was not reported in the final results. Transfection of both TR cDNAs resulted in cell surface localization of TR together with CD3, suggesting a productive protein in both cases. Tumor recognition of the Homo sapiens (Homsap) TRDV1-containing TR could be demonstrated by IFN Gamma ELISA ELISpot kit, whereas the Mus musculus (Musmus) TR did not recognize a tumor-derived cell line. To determine whether the TRDV-containing TRA chains we detected were rare events or whether TRDV genes are commonly incorporated into TRA chains, we queried the NCBI Sequence Read Archive for TR single-cell RNA-seq data and analyzed 21 human and 23 murine datasets. We found that especially Homsap TRDV1, Musmus TRDV1, and to some extent Musmus TRDV2-2 are more commonly incorporated into TRA chains than several TRAV genes, making those TRDV genes a relevant contribution to TRA diversity. TRDV-containing TRA chains are currently excluded from the final results of V-(D)-J dataset analyses with the CellRanger software. We provide a work-around to avoid exclusion of those hybrid TRA chains from the final analysis results.


New management strategies in the treatment of status epilepticus.
Status epilepticus is a neurologic emergency associated with high mortality and long-term disability. Recent advances in our understanding of the pathophysiological mechanisms involved in the initiation and perpetuation of seizure activity have revealed that status epilepticus is a dynamic and evolving process. Alterations at the cellular level parallel physiological, physical, and electrical changes at the bedside. Loss of cerebral autoregulation and neuronal damage begin after 30 minutes of continuous seizure activity. This understanding has led to changes in treatments of status epilepticus, which must be multidisciplinary and occur simultaneously in many different areas. The goals of pharmacological therapy are to terminate seizures early and prevent recurrence. Two recent large clinical studies have shown the benefit of early administration of benzodiazepines to control status epilepticus. Pharmacological algorithms designed to focus medical management have trended toward earlier and more aggressive treatment. The hope is that continued exploration into the basic mechanisms involved in status epilepticus and future controlled clinical trials defining optimal medical management will produce further advances.


Clinical Significance of Staphylococcus aureus in a Single Positive Blood Culture Bottle.
Clinical significance of a single positive blood culture bottle (SPBCB) with Staphylococcus aureus is unclear. We aimed to assess the significance of an SPBCB by looking at the associated outcomes. We performed a retrospective, multicenter study of patients with an SPBCB with S aureus using data collected from both electronic health records and the clinical microbiology laboratory. Overall, 534 patients with S aureus bacteremia were identified and 118 (22.1%) had an SPBCB. Among cases with an SPBCB, 106 (89.8%) were classified as clinically significant whereas 12 (10.2%) were considered contaminated or of unclear significance. A majority (92.4%) of patients received antibiotic therapy, but patients with clinically significant bacteremia were treated with longer courses (25.9 vs 5.7 days, P < .001). Significant differences in both frequency of echocardiography (65.1% vs 84.6%, P < .001) and infective endocarditis diagnosis (3.8% vs 14.2%, P = .002) were seen in those with an SPBCB compared to those with multiple positive bottles. A longer hospital length of stay and higher 90-day, 6-month, and 1-year mortality rates were seen in patients with multiple positive blood culture bottles. An SPBCB with S aureus was common among our patients. While this syndrome has a more favorable prognosis as compared to those with multiple positive blood cultures, clinicians should remain concerned as it portends a risk of infective endocarditis and mortality.


Investigation of the CADM2 polymorphism rs17518584 in memory and executive functions measures in a cohort of young healthy individuals.
The common polymorphism rs17518584, near the cell adhesion molecule 2 gene (CADM2), was previously identified as playing a role in information processing speed in a genome-wide association study of executive functions and processing speed performed in a cohort of non-demented older adults. In this study, we investigated this polymorphism in a younger population cohort (≤30 years old, median age 19 years), with no known memory or psychiatric disorders, for which we had phenotyped all participants for memory function (n = 514), and a subset of the participants for executive functions (n = 338), using a battery of tests measuring visuo-spatial memory, working memory, verbal memory, and frontal lobe functions (visual scanning, graphomotor speed, and cognitive flexibility). The polymorphism rs17518584 was genotyped by a restriction fragment length polymorphism assay and analysis indicated that the CADM2 polymorphism showed evidence of association with information processing speed as inferred from scores from the Stroop Word, Colour, and Colour-Word Tests (p = 0.005, p = 0.04, and p = 0.028, respectively, in a dominant inheritance model), as well as Trail Making Test Part A (p = 0.005 in an additive model). Significant associations of rs17518584 with scores from other tests of memory subtypes were not detected. The findings of this study provide further support for a role of CADM2 in aspects of cognitive function, in particular reading and information processing speed, and suggest that this role extends to younger individuals.


Sensitivity of marine microalgae to copper: the effect of biotic factors on copper adsorption and toxicity.
Microalgae are sensitive indicators of environmental change and, as the basis of most freshwater and marine ecosystems, are widely used in the assessment of risk and development of environmental regulations for metals. However, interspecies differences in sensitivity to metals are not well understood. The relationship between metal-algal cell binding and copper sensitivity of marine microalgae was investigated using a series of 72-h growth-rate inhibition bioassays and short-term (1-h) uptake studies. A range of marine algae from different taxonomic groups were screened to determine whether copper adsorption to the cell membrane was influenced by biotic factors, such as the ultrastructure of cell walls and cell size. Minutocellus polymorphus was the most sensitive species to copper and Dunaliella tertiolecta the least sensitive, with 72-h IC50 values (concentration to inhibit growth-rate by 50%) of 0.6 and 530 microg Cu/L, respectively. Copper solution-cell partition coefficients at equilibrium (K(d)) were calculated for six species of algae on a per cell and surface area basis. The largest and smallest cells had the lowest and highest K(d) values, respectively (on a surface area basis), with a general (non-linear) trend of decreasing K(d) with increasing cell surface area (p=0.026), however, no relationship was found between K(d) and copper sensitivity, nor cell size and copper sensitivity. Interspecies differences in copper sensitivity were not related to cell size, cell wall type, taxonomic group or K(d) values. The differences in sensitivity may be due to differences in uptake rates across the plasma membrane, in internal binding mechanisms and/or detoxification mechanisms between the different microalgal species.


Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production.
In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics. Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields. In this study, fluorescein-labelled monoclonal antibodies against influenza A virus matrix protein 1 and nucleoprotein were used for monitoring the infection status of adherent Madin-Darby canine kidney cells from bioreactor samples. Monoclonal antibody binding was shown for influenza A virus strains of different subtypes (H1N1, H1N2, H3N8) and host specificity (human, equine, swine). At high multiplicity of infection in a bioreactor, the onset of viral protein accumulation in adherent cells on microcarriers was detected at about 2 to 4 h post infection by flow cytometry. In contrast, a significant increase in titre by hemagglutination assay was detected at the earliest 4 to 6 h post infection. It is shown that flow cytometry is a sensitive and robust method for the monitoring of viral infection in fixed cells from bioreactor samples. Therefore, it is a valuable addition to other detection methods of influenza virus infection such as immunotitration and RNA hybridisation. Thousands of individual cells are measured per sample. Thus, the presented method is believed to be quite independent of the concentration of infected cells (multiplicity of infection and total cell concentration) in bioreactors. This allows to perform detailed studies on factors relevant for optimization of virus yields in cell cultures. The method could also be used for process characterisation and investigations concerning reproducibility in vaccine manufacturing.


Genetic isolation between two sympatric host-plant races of the European corn borer, Ostrinia nubilalis Hübner. I. Sex pheromone, moth emergence timing, and parasitism.
Adaptation to different environments may be a powerful source of genetic differentiation between populations. The biological traits selected in each environment can pleiotropically induce assortative mating between individuals of these genetically differentiated populations. This situation may facilitate sympatric speciation. Successful host shifts in phytophagous insects provide some of the best evidence for the ecological speciation that occurs, or has occurred, in sympatry. The European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), colonized maize after its introduction into Europe by humans about 500 years ago. In northern France, two sympatric host races feed on maize (Zea mays) and mugwort (Artemisia vulgaris), respectively. We investigated the factors involved in the genetic isolation of these two races at a field site near Paris, France. We identified two biological differences that might make a significant contribution to the genetic divergence between sympatric populations feeding on the two host plants. First, assortative mating may be due to differences in the moth emergence pattern between the two races: mugwort-race moths emerged on average 10 days earlier than maize-race moths. In addition, the males emerged earlier than females in both races. Hence, the likelihood of mating between maize-race males and mugwort-race females was higher than that of mating between mugwort-race males and maize-race females. Second, the females feeding on mugwort and maize produced sex pheromones with different E/Z isomeric ratios of delta-11-tetradecenyl acetate. This difference in mate recognition systems reinforces the potential for assortative mating in the two races. During the experiment, overwintering mortality was much lower on maize than on mugwort. This difference was due to a braconid parasitoid wasp, Macrocentrus cingulum, that killed more than 50% of the larvae overwintering on mugwort but did not infest larvae diapausing on maize. Hence, by colonizing maize, European corn borer populations probably escaped from numerous predators, competitors, and parasitoids, such as M. cingulum. This decrease in host-associated selection may have favored the colonization of this new host. Finally, throughout this experiment we observed selection at two allozyme loci (or at linked loci): Tpi and Mpi. The Tpi locus is tightly linked with the genes involved in the response of the male to the sex pheromone and in developmental timing. The location of these traits on the Z chromosome may play a role in shortening the time required for the evolution of premating barriers.


The effect of aqueous extracts of Hibiscus sabdariffa (Sorrel) calyces on heamatological profile and organ pathological changes in Trypanasoma congolense - infected rats.
The effects of aqueous extract of Hibiscus sabdariffa calyces on haematology and pathological changes in some selected organs during experimental Trypanosoma congolense infection of rats were investigated. Three groups of rats were intraperitoneally infected with T. congolense (Karu stock). One group was administered with the aqueous extract and another given a solution of vitamin C in drinking water; the remaining infected group was left untreated. Data from these groups were compared with those of two groups of healthy rats, one of which was similarly treated with the aqueous extract. The experiment was terminated three weeks, post-infection (pi). The uninfected and infected rats administered the extract consumed the equivalent of 9.94 mg - and 9.61 mg ascorbic acid / 100g / day during the experiment. Consumption of the extract significantly (p<0.01) retarded the rate of weight gain in both healthy and infected rats; even though the feed-intake was not significantly affected. After two weeks of infection the extract and vitamin C kept the parasitaemia significantly (p<0.01) lower than the untreated infected group. The anaemia in the untreated infected group was significantly (p<0.01) more severe than that of the corresponding extract- or vitamin-treated groups. Trypanosoma congolense infection caused significant (p<0.01) decreases in serum total proteins and albumin; serum and organ ascorbic acid as well as significant (p<0.01) elevation of serum alanine amino transferase levels in untreated rats. Consumption of the extract or vitamin C, however, prevented these disease-induced anomalies in the treated infected rats. Serum creatinine and urea levels were not affected by infection but the extract elevated these parameters significantly (p<0.01) above infection levels. It was concluded that consumption of the extract ameliorated the pathological changes in blood and organs of T. congolense-infected rats.


[Activated protein C resistance as a cause of thrombophilia].
The proportion of identifiable causes of familial thrombophilia has increased from 5-10% to 60-70% since the identification of activated protein C resistance (aPCR) in February 1993 by Dahlbäck et al. A mutation in the factor V gene (G-->A, 1691) leads to the so called Leiden mutation (R 506 Q) that produces a mutated factor V resistant to the catalytic action of activated protein C (aPC), yet normal in its procoagulant properties. This recently identified aPCR is in Nordic populations the most prevalent and well defined genetic defect associated with disease so far described. Its prevalence in the general population ranges from 0% to up to 15% and suggests that a positive genetic selection pressure has been involved. The aPCR phenotype can be assessed in vitro by measurement of the prolongation of the activated partial thromboplastin time in the presence of aPC, whereas the aPCR genotype is studied using polymerase chain reaction searching for the Arg to Gln mutation in the coagulation factor V gene. Some acquired conditions such as the presence of lupus anticoagulants, antiphospholipid antibodies, pregnancy, liver disease and contraceptives may lead into the aPCR phenotype. The aPCR search must be the initial step in the study of a patient with thrombophilia, either inherited or acquired aPCR together with protein C, protein S and antithrombin III explain 60 to 70% of cases of familial thrombophilia.