Habit learning in Tourette syndrome: a translational neuroscience approach to a developmental psychopathology.
The etiology of Tourette syndrome (TS) involves disturbances in the structure and function of the basal ganglia. The basal ganglia mediate habit learning. To study habit learning in persons with TS. Patients with TS were compared with normal controls in performance on a probabilistic classification, or habit-learning task (weather prediction). University research institute. One hundred twenty-three children and adults, 56 with a diagnosis of TS and 67 healthy control subjects. Habit learning was assessed by the extent of improvement in accuracy of predictions and reaction times over trial blocks during performance of the weather prediction task. Declarative learning was assessed by performance on 3 tasks that required intact declarative memory functioning. Children with TS were impaired at habit learning relative to normal controls (P = .01). This finding was replicated in the independent sample of adults with TS (P = .01). The rate of learning correlated inversely with the severity of tic symptoms across both samples (r = -0.34; P = .01). Thus, impaired learning accompanied more severe symptoms. Measures of declarative memory functioning, in contrast, were normal in the TS groups. Striatal learning systems are uniquely dysfunctional in both children and adults with TS. The correlation of habit learning with symptom severity suggests that the number and severity of tics are a function of the degree to which the system for habit learning is dysfunctional. Thus, both the deficits in habit learning and the tic symptoms of TS are likely to be consequences of the previously reported anatomical and functional disturbances of the striatum in children and adults who have TS. The existence of a well-developed animal model for this learning system, which permits study of the neural and molecular bases of habit learning, has important implications for the neurobiological study of TS and for the development of new or improved therapeutics for this condition.


[The effect of hyperbaric oxygenation on central hemodynamics, blood gas composition and acid-base status in elderly patients with ischemic heart disease and circulatory insufficiency].
210 patients with ischemic heart disease and heart failure, stage I, HA, IIB, aged 60 to 74 years, have been examined. The effect of HBO on central hemodynamics, gas composition and acid-base balance has been assessed. It has been established that HBO is associated with a definite tendency towards cardiac output normalization, as well as hemodynamic economization associated with accelerated heart rate and more adequate correlation between cardiac output and peripheral vascular resistance. The analysis of gas exchange and acid-base balance parameters indicates HBO-induced acidotic shifts. The data of polarographic studies demonstrate that increased metabolic activity is typical not only of the cardiac muscle but of other tissues as well. A favourable effect of HBO on oxygen tissue supply in patients with ischemic heart disease and heart failure is to a certain extent due to stimulation of peripheral circulation and its capillary compartment.


Susceptibility of five different sizes of pathogenfree Litopenaeus vannamei to white spot syndrome virus (WSSV) by intramuscular inoculation.
White spot syndrome virus (WWSV) has become one of the most widespread causes of mortality in commercial shrimp farming. In the present study, we used PCR to determine the shrimp infectious dose 50% endpoint (SID50 ml-1) of a Chinese isolate of WSSV in 5 different sizes of pathogen-free Litopenaeus vannamei inoculated intramuscularly. The lethal dose 50% endpoint (LD50 ml-1) was also determined from the percentage of dead shrimp. The LD50 ml-1 for 2, 4, 6, 8, and 10 cm shrimp were 104.68, 105.7, 106.70, 107.75, and 108.81, respectively, and the SID50 ml-1 were 104.68, 105.70, 106.90, 107.75, and 108.94, respectively. There was no significant difference between the LD50 ml-1 and SID50 ml-1 for each shrimp size, which indicated that all infected shrimp died. The lethal and infectious titer decreased about 1 log10 as shrimp size decreased 1 grade. These data clearly indicate that adult shrimp were more susceptible to WSSV than juvenile shrimp. The horizontal comparison showed that the amount of virus in the shrimp organs increased over the experimental period. The vertical comparison showed that virus quantity was lowest in the organs of 10 cm shrimp and highest in 2 cm shrimp, which indicates that the smaller shrimp had higher levels of viral replication. Hence, the optimal size for WSSV challenge in shrimp inoculated intramuscularly was 2 cm. The determination of virus titers in different sizes of shrimp represents a step towards creating strategies to reduce the negative impacts of WSSV in the aquaculture industry.


SP1 and NFY Regulate the Expression of PNPT1, a Gene Encoding a Mitochondrial Protein Involved in Cancer.
The Polyribonucleotide nucleotidyltransferase 1 gene (PNPT1) encodes polynucleotide phosphorylase (PNPase), a 3'-5' exoribonuclease involved in mitochondrial RNA degradation and surveillance and RNA import into the mitochondrion. Here, we have characterized the PNPT1 promoter by in silico analysis, luciferase reporter assays, electrophoretic mobility shift assays (EMSA), chromatin immunoprecipitation (ChIP), siRNA-based mRNA silencing and RT-qPCR. We show that the Specificity protein 1 (SP1) transcription factor and Nuclear transcription factor Y (NFY) bind the PNPT1 promoter, and have a relevant role regulating the promoter activity, PNPT1 expression, and mitochondrial activity. We also found in Kaplan-Meier survival curves that a high expression of either PNPase, SP1 or NFY subunit A (NFYA) is associated with a poor prognosis in liver cancer. In summary, our results show the relevance of SP1 and NFY in PNPT1 expression, and point to SP1/NFY and PNPase as possible targets in anti-cancer therapy.


Eradication of HIV by transplantation of CCR5-deficient hematopoietic stem cells.
Today, 30 years after the onset of the HIV pandemic, although treatment strategies have considerably improved, there is still no cure for the disease. Recently, we described a successful hematopoietic stem cell transplantation in an HIV-1-infected patient, transferring donor-derived cells with a natural resistance against HIV infection. These hematopoietic stem cells engrafted, proliferated, and differentiated into mature myeloid and lymphoid cells. To date, the patient has not required any antiretroviral treatment, more than 4 years after allogeneic transplantation. In the analysis of peripheral blood cells and different tissue samples, including gut, liver, and brain, no viral load or proviral DNA could be detected. Our report raises the hope for further targeted treatment strategies against HIV and represents a successful personalized treatment with allogeneic stem cells carrying a beneficial gene. However, this case has ignited a controversy regarding the question of whether this patient has achieved complete eradication of HIV or not. Here we give an update on open questions, unsolved aspects, and clinical consequences concerning this unique case.


Confidentially and the family physician.
The principle of confidentiality is one of the clearest and firmest rules of professional ethics. However, situations can arise in which confidentiality ought to be abridged in favor of other, weightier values. In the case of a teenager who asks a family physician to prescribe birth control pills for her without informing her parents, the best interests of the family as a unit may justify a certain aggressive but oblique strategy abridging confidentiality.


Polychlorinated biphenyl (PCB) exposure in relation to thyroid hormone levels in neonates.
Polychlorinated biphenyls (PCBs) are industrially produced environmentally persistent compounds. In developed countries all humans have detectable levels in blood and other tissues. PCBs alter thyroid hormone metabolism in animal experiments, and human data suggest background-level exposure may have similar effects in neonates. We evaluated this possible effect among 160 North Carolina children whose in utero PCB exposure was estimated on the basis of the mother's PCB levels in milk and blood, in 1978-1982 (estimated median PCB level in milk at birth, 1.8 mg/kg lipid). Their umbilical cord sera were thawed in 1998 and assayed for total thyroxine, free thyroxine, and thyroid stimulating hormone. We found that PCB exposure was not strongly related to any of the thyroid measures. For example, for a one unit change in milk PCB concentration (mg/kg lipid), the associated multivariate-adjusted increase in thyroid stimulating hormone level was 7% (95% confidence limits (CL) = -6, 21). Despite the possibility of sample degradation, these data suggest that within the range of background-level exposure in the United States, in utero PCB exposure is only slightly related to serum concentration of total thyroxine, free thyroxine, and thyroid stimulating hormone at birth.


The Relationship Between Acute: Chronic Workload Ratios and Injury Risk in Sports: A Systematic Review.
Low injury rates have previously been correlated with sporting team success, highlighting the importance of injury prevention programs. Recent methods, such as acute:chronic workload ratios (ACWR) have been developed in an attempt to predict and manage injury risk; however, the relation between these methods and injury risk is unclear. The aim of this systematic review was to identify and synthesize the key findings of studies that have investigated the relationship between ACWR and injury risk. Included studies were critically appraised using the Downs and Black checklist, and a level of evidence was determined. Relevant data were extracted, tabulated, and synthesized. Twenty-seven studies were included for review and ranged in percentage quality scores from 48.2% to 64.3%. Almost perfect interrater agreement (κ = 0.885) existed between raters. This review found a high variability between studies with different variables studied (total distance versus high speed running), as well as differences between ratios analyzed (1.50-1.80 versus ≥1.50), and reference groups (a reference group of 0.80-1.20 versus ≤0.85). Considering the high variability, it appears that utilizing ACWR for external (eg, total distance) and internal (eg, heart rate) loads may be related to injury risk. Calculating ACWR using exponentially weighted moving averages may potentially result in a more sensitive measure. There also appears to be a trend towards the ratios of 0.80-1.30 demonstrating the lowest risk of injury. However, there may be issues with the ACWR method that must be addressed before it is confidently used to mitigate injury risk. Utilizing standardized approaches will allow for more objective conclusions to be drawn across multiple populations.


TLR4 and Insulin Resistance.
Chronic inflammation is a key feature of insulin resistance and obesity. Toll-Like Receptor 4 (TLR4), involved in modulating innate immunity, is an important mediator of insulin resistance and its comorbidities. TLR4 contributes to the development of insulin resistance and inflammation through its activation by elevated exogenous ligands (e.g., dietary fatty acids and enteric lipopolysaccharide) and endogenous ligands (e.g., free fatty acids) which are elevated in obese states. TLR4, expressed in insulin target tissues, activates proinflammatory kinases JNK, IKK, and p38 that impair insulin signal transduction directly through inhibitory phosphorylation of insulin receptor substrate (IRS) on serine residues. TLR4 activation also leads to increased transcription of pro-inflammatory genes, resulting in elevation of cytokine, chemokine, reactive oxygen species, and eicosanoid levels that promote further insulin-desensitization within the target cell itself and in other cells via paracrine and systemic effects. Increased understanding of cell type-specific TLR4-mediated effects on insulin action present the opportunity and challenge of developing related therapeutic approaches for improving insulin sensitivity while preserving innate immunity.


The GPR139 reference agonists 1a and 7c, and tryptophan and phenylalanine share a common binding site.
GPR139 is an orphan G protein-coupled receptor expressed in the brain, in particular in the habenula, hypothalamus and striatum. It has therefore been suggested that GPR139 is a possible target for metabolic disorders and Parkinson's disease. Several surrogate agonist series have been published for GPR139. Two series published by Shi et al. and Dvorak et al. included agonists 1a and 7c respectively, with potencies in the ten-nanomolar range. Furthermore, Isberg et al. and Liu et al. have previously shown that tryptophan (Trp) and phenylalanine (Phe) can activate GPR139 in the hundred-micromolar range. In this study, we produced a mutagenesis-guided model of the GPR139 binding site to form a foundation for future structure-based ligand optimization. Receptor mutants studied in a Ca2+ assay demonstrated that residues F1093×33, H1875×43, W2416×48 and N2717×38, but not E1083×32, are highly important for the activation of GPR139 as predicted by the receptor model. The initial ligand-receptor complex was optimized through free energy perturbation simulations, generating a refined GPR139 model in agreement with experimental data. In summary, the GPR139 reference surrogate agonists 1a and 7c, and the endogenous amino acids L-Trp and L-Phe share a common binding site, as demonstrated by mutagenesis, ligand docking and free energy calculations.


Expression of basic fibroblast growth factor is necessary but insufficient for production of metastasis.
We determined whether overexpression of basic fibroblast growth factor (bFGF) is necessary for enhanced growth and production of metastasis by murine K-1735 melanoma cells. The bFGF gene was transfected into three nonmetastatic clones (C-IO, C-19, and C-23) that do not express bFGF mRNA and protein and one metastatic clone that expresses high levels of bFGF mRNA and protein. Control cells were transfected with a dominant selectable marker neomycin resistance gene (neo). All bFGF-transduced cells expressed bFGF-specific mRNA transcripts and cellular bFGF protein and proliferated in culture with medium containing low concentrations of serum. Anchorage-independent growth in hard agarose was enhanced only in bFGF-transfected nonmetastatic C-10 cells which, subsequent to the transfection, also expressed high levels of collagenase IV/gelatinase A activity. The treatment of C-10, C-19, and C-23 cells with exogenous bFGF induced collagenase IV/gelatinase expression, as did the addition of lysates from C-10/bFGF and C-23/bFGF cells. C-10/bFGF cells (but not C-19/bFGF or C-23/bFGF) produced highly vascular and rapidly growing subcutaneous tumors as well as a high incidence of lung metastasis. These data suggest that overexpression of bFGF is necessary but in itself not sufficient to convert nonmetastatic K-1735 cells to the metastatic phenotype and that enhanced tumorigenicity and metastasis require at least concurrent expression of bFGF and collagenase type TV genes.


Small-molecule inhibitors of bacterial AddAB and RecBCD helicase-nuclease DNA repair enzymes.
The AddAB and RecBCD helicase-nucleases are related enzymes prevalent among bacteria but not eukaryotes and are instrumental in the repair of DNA double-strand breaks and in genetic recombination. Although these enzymes have been extensively studied both genetically and biochemically, inhibitors specific for this class of enzymes have not been reported. We developed a high-throughput screen based on the ability of phage T4 gene 2 mutants to grow in Escherichia coli only if the host RecBCD enzyme, or a related helicase-nuclease, is inhibited or genetically inactivated. We optimized this screen for use in 1536-well plates and screened 326,100 small molecules in the NIH molecular libraries sample collection for inhibitors of the Helicobacter pylori AddAB enzyme expressed in an E. coli recBCD deletion strain. Secondary screening used assays with cells expressing AddAB or RecBCD and a viability assay that measured the effect of compounds on cell growth without phage infection. From this screening campaign, 12 compounds exhibiting efficacy and selectivity were tested for inhibition of purified AddAB and RecBCD helicase and nuclease activities and in cell-based assays for recombination; seven were active in the 0.1-50 μM range in one or another assay. Compounds structurally related to two of these were similarly tested, and three were active in the 0.1-50 μM range. These compounds should be useful in further enzymatic, genetic, and physiological studies of these enzymes, both purified and in cells. They may also lead to useful antibacterial agents, since this class of enzymes is needed for successful bacterial infection of mammals.


Surgical and percutaneous myocardial angiogenesis induction. Part II--neoangiogenesis.
Coronary artery bypass surgery and angioplasty provide symptomatic relief in patients with ischemic heart disease, but despite advancement in technique and devices, these methods are not applicable to a subset of patients with angina refractory to medical treatment. Bypass surgery might not be feasible because of lack of suitable conduits, diffuse coronary disease or poor distal run-off, and coronary angioplasty is sometimes not applicable due to chronic total occlusion, diffuse disease or extreme tortuosity. We have previously reviewed the available experience with laser-induced direct myocardial revascularization, one of the new potential treatment modalities for this patient subset. One of the potential mechanisms of action for laser treatment is the induction of neoangiogenesis. In the second part of our article we review the available experience with the induction of myocardial angiogenesis using different growth factors or the genes encoding for them.


Use of Simulation Based on an Electronic Health Records Environment to Evaluate the Structure and Accuracy of Notes Generated by Medical Scribes: Proof-of-Concept Study.
The increasing adoption of electronic health records (EHRs) has been associated with a number of unintended negative consequences with provider efficiency and job satisfaction. To address this, there has been a dramatic increase in the use of medical scribes to perform many of the required EHR functions. Despite this rapid growth, little has been published on the training or assessment tools to appraise the safety and efficacy of scribe-related EHR activities. Given the number of reports documenting that other professional groups suffer from a number of performance errors in EHR interface and data gathering, scribes likely face similar challenges. This highlights the need for new assessment tools for medical scribes. The objective of this study was to develop a virtual video-based simulation to demonstrate and quantify the variability and accuracy of scribes' transcribed notes in the EHR. From a pool of 8 scribes in one department, a total of 5 female scribes, intent on pursuing careers in health care, with at least 6 months of experience were recruited for our simulation study. We created three simulated patient-provider scenarios. Each scenario contained a corresponding medical record in our simulation instance of our EHR. For each scenario, we video-recorded a standardized patient-provider encounter. Five scribes with at least 6 months of experience both with our EHR and in the specialty of the simulated cases were recruited. Each scribe watched the simulated encounter and transcribed notes into a simulated EHR environment. Transcribed notes were evaluated for interscribe variability and compared with a gold standard for accuracy. All scribes completed all simulated cases. There was significant interscribe variability in note structure and content. Overall, only 26% of all data elements were unique to the scribe writing them. The term data element was used to define the individual pieces of data that scribes perceived from the simulation. Note length was determined by counting the number of words varied by 31%, 37%, and 57% between longest and shortest note between the three cases, and word economy ranged between 23% and 71%. Overall, there was a wide inter- and intrascribe variation in accuracy for each section of the notes with ranges from 50% to 76%, resulting in an overall positive predictive value for each note between 38% and 81%. We created a high-fidelity, video-based EHR simulation, capable of assessing multiple performance indicators in medical scribes. In this cohort, we demonstrate significant variability both in terms of structure and accuracy in clinical documentation. This form of simulation can provide a valuable tool for future development of scribe curriculum and assessment of competency.


A new perspective on the importance of glycine conjugation in the metabolism of aromatic acids.
A number of endogenous and xenobiotic organic acids are conjugated to glycine, in animals ranging from mosquitoes to humans. Glycine conjugation has generally been assumed to be a detoxification mechanism, increasing the water solubility of organic acids in order to facilitate urinary excretion. However, the recently proposed glycine deportation hypothesis states that the role of the amino acid conjugations, including glycine conjugation, is to regulate systemic levels of amino acids that are also utilized as neurotransmitters in the central nervous systems of animals. This hypothesis is based on the observation that, compared to glucuronidation, glycine conjugation does not significantly increase the water solubility of aromatic acids. In this review it will be argued that the major role of glycine conjugation is to dispose of the end products of phenylpropionate metabolism. Furthermore, glucuronidation, which occurs in the endoplasmic reticulum, would not be ideal for the detoxification of free benzoate, which has been shown to accumulate in the mitochondrial matrix. Glycine conjugation, however, prevents accumulation of benzoic acid in the mitochondrial matrix by forming hippurate, a less lipophilic conjugate that can be more readily transported out of the mitochondria. Finally, it will be explained that the glycine conjugation of benzoate, a commonly used preservative, exacerbates the dietary deficiency of glycine in humans. Because the resulting shortage of glycine can negatively influence brain neurochemistry and the synthesis of collagen, nucleic acids, porphyrins, and other important metabolites, the risks of using benzoate as a preservative should not be underestimated.


Vascular access in a University transplant and dialysis program. Results, costs, and manpower implications.
Vascular access has become the most common operation performed at North Carolina Memorial Hospital, Chapel Hill. Three hundred vascular access procedures performed between 1971 and 1975 were reviewed as to results, costs, and manpower commitment. Subcutaneous radial arterial venous fistula was the procedure of choice, with a 67% two-year patency and the lowest incidence of complications. An average of 1.7 operations per patient was necessary to maintain access. Patients used an average of 19 hospital days per year of dialysis for vascular access and an average of 35 hospital days per year of dialysis for all medical problems. Cost of hospitalization averaged $6,818 per patient per year of dialysis, and for vascular access alone averaged $3,452 per patient per year of dialysis. To create and maintain vascular access, 1,000 hours of operating room time were required, at an expense of $200,000. Vascular access in support of patients undergoing chronic hemodialysis requires a massive commitment of dollars, time, and personnel.


Programmable bio-nanochip-based cytologic testing of oral potentially malignant disorders in Fanconi anemia.
Fanconi anemia (FA) is caused by mutations of DNA repair genes. The risk of oral squamous cell carcinoma (OSCC) among FA patients is 800-folds higher than in the general population. Early detection of OSCC, preferably at it precursor stage, is critical in FA patients to improve their survival. In an ongoing clinical trial, we are evaluating the effectiveness of the programmable bio-nanochip (p-BNC)-based oral cytology test in diagnosing oral potentially malignant disorders (OPMD) in non-FA patients. We used this test to compare cytomorphometric and molecular biomarkers in OSCC cell lines derived from FA and non-FA patients to brush biopsy samples of a FA patient with OPMD and normal mucosa of healthy volunteers. Our data showed that expression patterns of molecular biomarkers were not notably different between sporadic and FA-OSCC cell lines. The p-BNC assay revealed significant differences in cytometric parameters and biomarker MCM2 expression between cytobrush samples of the FA patient and cytobrush samples of normal oral mucosa obtained from healthy volunteers. Microscopic examination of the FA patient's OPMD confirmed the presence of dysplasia. Our pilot data suggests that the p-BNC brush biopsy test recognized dysplastic oral epithelial cells in a brush biopsy sample of a FA patient.


Versatile roles of V-ATPases accessory subunit Ac45 in osteoclast formation and function.
Vacuolar-type H(+)-ATPases (V-ATPases) are macromolecular proton pumps that acidify intracellular cargos and deliver protons across the plasma membrane of a variety of specialized cells, including bone-resorbing osteoclasts. Extracellular acidification is crucial for osteoclastic bone resorption, a process that initiates the dissolution of mineralized bone matrix. While the importance of V-ATPases in osteoclastic resorptive function is well-defined, whether V-ATPases facilitate additional aspects of osteoclast function and/or formation remains largely obscure. Here we report that the V-ATPase accessory subunit Ac45 participates in both osteoclast formation and function. Using a siRNA-based approach, we show that targeted suppression of Ac45 impairs intracellular acidification and endocytosis, both are prerequisite for osteoclastic bone resorptive function in vitro. Interestingly, we find that knockdown of Ac45 also attenuates osteoclastogenesis owing to a reduced fusion capacity of osteoclastic precursor cells. Finally, in an effort to gain more detailed insights into the functional role of Ac45 in osteoclasts, we attempted to generate osteoclast-specific Ac45 conditional knockout mice using a Cathepsin K-Cre-LoxP system. Surprisingly, however, insertion of the neomycin cassette in the Ac45-Flox(Neo) mice resulted in marked disturbances in CNS development and ensuing embryonic lethality thus precluding functional assessment of Ac45 in osteoclasts and peripheral bone tissues. Based on these unexpected findings we propose that, in addition to its canonical function in V-ATPase-mediated acidification, Ac45 plays versatile roles during osteoclast formation and function.


[The effect of prolonged exposure to heat on the activity of salivary gland polytene chromosomes].
The influence of long-term heating on the puffing activity of polytene chromosomes in the early prepupa salivary glands was investigated. The activity of puffs was estimated by two criteria: size and frequency. The rearing of insects at a temperature of 29 degrees resulted in puff changes: the activity of some puffs increased or depressed, some puffs were inhibited, other puffs were induced newly. The differential response of each chromosome was observed. A possible mechanism of the effect of heating on the puff activity of polytene chromosomes is discussed.


Climate and agricultural context shape reproductive mode variation in an aphid crop pest.
In aphids, reproductive mode is generally assumed to be selected for by winter climate. Sexual lineages produce frost-resistant eggs, conferring an advantage in regions with cold winters, while asexual lineages predominate in regions with mild winters. However, habitat and resource heterogeneities are known to exert a strong influence on sex maintenance and might modulate the effect of climate on aphid reproductive strategies. We carried out a hierarchical sampling in northern France to investigate whether reproductive mode variation of the aphid Rhopalosiphum padi is driven by winter climate conditions, by habitat and resource heterogeneities represented by a range of host plants or by both factors. We confirmed the coexistence in R. padi populations of two genetic clusters associated with distinct reproductive strategies. Asexual lineages predominated, whatever the surveyed year and location. However, we detected a between-year variation in the local contribution of both clusters, presumably associated with preceding winter severity. No evidence for host-driven niche differentiation was found in the field on six Poaceae among sexual and asexual lineages. Two dominant multilocus genotypes ( approximately 70% of the sample), having persisted over a 10-year period, were equally abundant on different plant species and locations, indicating their large ecological tolerance. Our results fit theoretical predictions of the influence of winter climate on the balance between sexual and asexual lineages. They also highlight the importance of current agricultural practices which seem to favour a small number of asexual generalist genotypes and their migration across large areas of monotonous environments.


More time for complex consultations in a high-deprivation practice is associated with increased patient enablement.
Evidence of the beneficial effects of longer consultations in general practice is limited. To evaluate the effect of increasing consultation length on patient enablement in general practice in an area of extreme socioeconomic deprivation. Longitudinal study using a 'before and after' design. Keppoch Medical Centre in Glasgow, which serves the most deprived practice area in Scotland. Participants were 300 adult patients at baseline, before the introduction of longer consultations, and 324 at follow-up, more than 1 year after the introduction of longer consultations. The intervention studied was more time in complex consultations. Patient satisfaction, perceptions of the GPs' empathy, GP stress, and patient enablement were collected by face-to-face interview. Additional qualitative data were obtained by individual interviews with the GPs, relating to their perceptions of the impact of the longer consultations. Response rates of 70% were obtained. Overall, 53% of consultations were complex. GP stress was higher in complex consultations. Patient satisfaction and perception of the GPs' empathy were consistently high. Average consultation length in complex consultations was increased by 2.5 minutes by the intervention. GP stress in consultations was decreased after the introduction of longer consultations, and patient enablement was increased. GPs' views endorsed these findings, with more anticipatory and coordinated care being possible in the longer consultations. More resource to provide more time in complex consultations in an area of extreme deprivation is associated with an increase in patient enablement.


Implication of human drug transporters to toxicokinetics and toxicity of pesticides.
Human membrane drug transporters are recognized as major actors of pharmacokinetics. Pesticides also interact with human drug transporters, which may have consequences for pesticide toxicokinetics and toxicity. The present review summarizes key findings about this topic. In vitro assays have demonstrated that some pesticides, belonging to various chemical classes, modulate drug transporter activity, regulate transporter expression and/or are substrates, thus bringing the proof of concept for pesticide-transporter relationships. The expected low human concentration of pesticides in response to environmental exposure constitutes a key-parameter to be kept in mind for judging the in vivo relevance of such pesticide-transporter interactions and their consequences for human health. Existing data about interactions of pesticides with drug transporters remain, however, rather sparse; more extensive and systematic characterization of pesticide-transporter relationships, through the use of high throughput in vitro assays and/or in silico methods, is, therefore, required. In addition, consideration of transporter polymorphisms, pesticide mixture effects and physiological and pathological factors governing drug transporter expression may help to better define the in vivo relevance of pesticide-transporter interactions in terms of toxicokinetics and toxicity for humans. © 2019 Society of Chemical Industry.


Surgical Treatment for Patients with Post-traumatic Flexion Contracture of Proximal Interphalangeal Joint: Analysis of Various Affecting Factors.
Background: A flexion contracture (FC) of the proximal interphalangeal (PIP) joint can have a profound negative influence on daily activity. The outcomes of surgical release of the PIP joint in literature are based on small sample size studies done several decades ago. The aim of this study is to report the outcomes of surgical treatment for post-traumatic FC of the PIP joint and to identify factors that affect these outcomes. Methods: This single institute retrospective study included patients from 2000 to 2020. We only included patients with post-traumatic FC of the PIP joint. We evaluated the demographic characteristics, cause of FC, surgical approaches and the various procedures conducted. We surveyed postoperative complications. During the study period, we asked about their current symptoms and evaluated their operative outcomes as excellent, good, fair or poor through the phone. Results: The average FC recovery angle was 37.3°. The small finger was the most affected, and the most common cause of FC was a tendon laceration. The volar plate complex release was the most frequently conducted procedure. The FC improvement was positively correlated to the degree of preoperative FC. The more severe preoperative flexion-extension arc was presented, the more FC recovery was achieved after operation. Patients who underwent multiple procedures had a higher degree of preoperative FC, and better correction was achieved with multiple procedures than with a single procedure. The most critical complication was recurrence. Conclusions: We were able to obtain average 37.3° of extension by surgical treatment. The more severe the FC presented before surgery, the greater the need for multiple procedures, however, this resulted in a significant increase in joint extension. Nevertheless, caution should be exercised regarding recurrence and could occur even with an experienced surgeon. Level of Evidence: Level IV (Therapeutic).


Advancing osmotic power generation by covalent organic framework monolayer.
Osmotic power, also known as 'blue energy', is produced by mixing solutions of different salt concentrations, and represents a vast, sustainable and clean energy source. The efficiency of harvesting osmotic power is primarily determined by the transmembrane performance, which is in turn dependent on ion conductivity and selectivity towards positive or negative ions. Atomically or molecularly thin membranes with a uniform pore environment and high pore density are expected to possess an outstanding ion permeability and selectivity, but remain unexplored. Here we demonstrate that covalent organic framework monolayer membranes that feature a well-ordered pore arrangement can achieve an extremely low membrane resistivity and ultrahigh ion conductivity. When used as osmotic power generators, these membranes produce an unprecedented output power density over 200 W m-2 on mixing the artificial seawater and river water. This work opens up the application of porous monolayer membranes with an atomically precise structure in osmotic power generation.


Protein pattern alterations in hippocampal and cortical cells as a function of training in rats.
The study reports changes in the protein pattern and incorporation of L-[U-14C]-leucine in brain cells of the hippocampus and sensorimotor cortex of rats. The following subcellular fractions were analyzed by SDS-acrylamide gel electrophoresis: plasma membranes, synaptosomal membranes and synaptic mitochondria. Recurring reversal training gave an increased synthesis of synaptosomal membrane proteins with mol. wt. 35,000-45,000 and 60,000 and 100,000 in trained animals compared to active controls. Lesser changes were observed in plasma membrane and synaptic mitochondria fractions. Of the brain areas studied, the hippocampal synaptosomal fraction showed an initial, temporary response, and the cortical cell fractions responded subsequently. Judged from the time sequence of the protein response, it seems that recurrent reversal training induces a change in synaptic protein towards higher molecular weights, suggesting that these changes reflect a modification of the distribution of synaptic protein.


Alzheimer's disease diagnostics by a 3D deeply supervised adaptable convolutional network.
Early diagnosis is playing an important role in preventing progress of the Alzheimer's disease (AD). This paper proposes to improve the prediction of AD with a deep 3D Convolutional Neural Network (3D-CNN), which can show generic features capturing AD biomarkers extracted from brain images, adapt to different domain datasets, and accurately classify subjects with improved fine-tuning method. The 3D-CNN is built upon a convolutional autoencoder, which is pre-trained to capture anatomical shape variations in structural brain MRI scans for source domain. Fully connected upper layers of the 3D-CNN are then fine-tuned for each task-specific AD classification in target domain. In this paper, deep supervision algorithm is used to improve the performance of already proposed 3D Adaptive CNN. Experiments on the ADNI MRI dataset without skull-stripping preprocessing have shown that the proposed 3D Deeply Supervised Adaptable CNN outperforms several proposed approaches, including 3D-CNN model, other CNN-based methods and conventional classifiers by accuracy and robustness. Abilities of the proposed network to generalize the features learnt and adapt to other domains have been validated on the CADDementia dataset.


Orthotopic urinary diversion after radical cystectomy in treatment of muscle invasive bladder cancer.
Surgical treatment of invasive carcinoma of the bladder in males includes total cystectomy removal of the prostate, seminal vesicles, and the distal parts of the urethers and the pelvic lymph node dissection as well. At this moment it is not possible to recommend a particular type of urinary diversion, but today in clinical practice commonly used derivative are ileal orthotopic neobladder as the continent one and ileal conduit as non-continent urinary diversion. Continent urinary diversion after radical cystectomy are the result of the application of technological innovation in surgery, but also knowledge, imagination and skill of well trained urologist. This type of operation significantly improves the quality of life in patients who underwent radical cystectomy, and the proposal is to operate whenever there is a possibility for this type of procedure. Also it is very important, during surgery to respect oncological principles, of complete removal of tumorous tissue and that the functional principle of ensur- ing that the patients have daytime and also nighttime continence later on after the surgery.


Distribution of selenoglucosinolates and their metabolites in Brassica treated with sodium selenate.
In Brassica species, hydrolysis of (methylthio)glucosinolates produces sulfur-containing aglycons which have demonstrated anticancer benefits. Selenized Brassicaceae contain (methylseleno)glucosinolates and their selenium-containing aglycons. As a prelude to biological testing, broccoli, cauliflower, and forage rape plants were treated with sodium selenate and their tap roots, stems, leaves, and florets analyzed for selenoglucosinolates and their Se aglycons. Two new selenoglucosinolates were identified: glucoselenoraphanin in broccoli florets and glucoselenonasturtiin in forage rape roots. A new aglycon, selenoberteroin nitrile, was identified in forage rape. The major selenoglucosinolates were glucoselenoerucin in broccoli, glucoselenoiberverin in cauliflower, and glucoselenoerucin and glucoselenoberteroin in forage rape roots. In broccoli florets, the concentrations of selenglucosinolates exceeded those of their sulfur analogues. Fertilization with selenium slightly reduced (methylthio)glucosinolates and aglycons in the roots, but increased them in the florets, the leaves, and sometimes the stems. These discoveries provide a new avenue for investigating how consumption of Brassica vegetables and their organoselenides may promote human health.


Universal Health Coverage and the Right to Health: From Legal Principle to Post-2015 Indicators.
Universal Health Coverage (UHC) is widely considered one of the key components for the post-2015 health goal. The idea of UHC is rooted in the right to health, set out in the International Covenant on Economic, Social, and Cultural Rights. Based on the Covenant and the General Comment of the Committee on Economic, Social, and Cultural Rights, which is responsible for interpreting and monitoring the Covenant, we identify 6 key legal principles that should underpin UHC based on the right to health: minimum core obligation, progressive realization, cost-effectiveness, shared responsibility, participatory decision making, and prioritizing vulnerable or marginalized groups. Yet, although these principles are widely accepted, they are criticized for not being specific enough to operationalize as post-2015 indicators for reaching the target of UHC. In this article, we propose measurable and achievable indicators for UHC based on the right to health that can be used to inform the ongoing negotiations on Sustainable Development Goals. However, we identify 3 major challenges that face any exercise in setting indicators post-2015: data availability as an essential criterion, the universality of targets, and the adaptation of global goals to local populations.


Long-term results and complications of the syringopleural shunting for treatment of syringomyelia: a clinical study.
The use of drains in the treatment of syringomyelia has a simple and immediate appeal. Syringopleural shunting in syringomyelia has produced good short-term results, but limited information is available on long-term effects. We analyzed the complications and long-term outcomes after syringopleural shunting for syringomyelia. Fourthy-four patients with large-sized syringomyelia underwent syringopleural shunting because of spinal cord compression between 1992 and 2010 in our clinic. Thirty-two patients had Chiari malformation type I (Group B), and 12 patients were associated with primary parenchymal cavitations (Group A). Their ages ranged from 14 to 71 years. Both craniovertebral decompression and syringopleural shunting were performed on 21 patients, whereas only syringopleural shunting was performed on another 21 patients. The follow-up period ranged from 1 year to 17 years (mean: 9.1 years). There was no operative mortality. Early postoperative MRI revealed that syringes of 43 patients had collapsed. There were 9 (20.5%) minor complications in 9 patients, including temporary neurological deficits (6), respiratory distress (2) and headache (1). Seven (15.9%) serious complications [permanent neurological deficit (1), shunt migration (2), shunt misplacement (1), spinal instability (1), tethering (1), CSF over drainage (1)] were seen in five patients. Four of them were treated with a secondary operation. Three patients (3/9; 33.3%) who were treated by syringopleural shunt alone (Group B2) required craniovertebral decompression, although the shunt was functional. During long-term follow-up, three patients stabilized, five patients (11.3%) developed a worse neurological condition, and two of these patients died 10 and 7 years after surgery. Of all patients, 88.6% showed significant clinical improvement. Although there were complications and failures, syringopleural shunting produced satisfactory results at long-term follow-up.


Cost analysis of the built environment: the case of bike and pedestrian trials in Lincoln, Neb.
We estimated the annual cost of bike and pedestrian trails in Lincoln, Neb, using construction and maintenance costs provided by the Department of Parks and Recreation of Nebraska. We obtained the number of users of 5 trails from a 1998 census report. The annual construction cost of each trail was calculated by using 3%, 5%, and 10% discount rates for a period of useful life of 10, 30, and 50 years. The average cost per mile and per user was calculated. Trail length averaged 3.6 miles (range = 1.6-4.6 miles). Annual cost in 2002 dollars ranged from 25,762 to 248,479 (mean = 124,927; median = 171,064). The cost per mile ranged from 5735 to 54,017 (mean = 35,355; median = 37,994). The annual cost per user was 235 (range = 83-592), whereas per capita annual medical cost of inactivity was 622. Construction of trails fits a wide range of budgets and may be a viable health amenity for most communities. To increase trail cost-effectiveness, efforts to decrease cost and increase the number of users should be considered.


Anterior polar cataracts in CS rats: a predictor of mature cataract formation.
The objective of this study was to characterize the morphology of the anterior opacities formed during recovery from posterior subcapsular cataract (PSC) in Royal College of Surgeons (RCS) rats. Lenses from RCS rats at 8 and 12 weeks postnatal (n = 14 and 12, respectively) were examined under a dissecting microscope for the presence of anterior opacities. Lenses with anterior opacities were fixed, embedded in epoxy resin, and sectioned along the optic axis for light microscopy (LM) and transmission electron microscopy (TEM). At eight weeks postnatal, 21.5% of animals (3/14) had anterior cataracts. Light microscopy of 1- to 2-microm-thick sections revealed an anomalous layer of material located at the epithelium-fiber interface, which was identified as a zone of liquefaction by TEM. Epithelial cells had minor structural defects but were not necrotic. Anterior portions of elongating and cortical fibers under the zone of liquefaction were undisrupted, whereas their posterior portions had numerous vacuoles. The anterior opacities were classified as anterior polar cataracts (APCs) based on the location and type of morphologic damage in the affected lenses. At twelve weeks postnatal, 25% of animals (3/12) had APCs that involved prominent vesiculation of the anterior cortex. Ultrastructural examination showed that large vesicles were located between and inside anterior fibers and that most extracellular spaces were abnormally widened. Posteriorly, internalization of the PSC by new fiber growth was disordered and displayed vesiculation and density variations. In the bow region, LM revealed minor structural irregularities that were identified as groups of apparently degenerating fibers by TEM. APCs in RCS rats are caused by degeneration of elongating fibers in the bow region and subsequent damage in the superficial anterior cortex. The percentage of animals with APCs (25%) was consistent with the percentage of animals in which mature cataracts eventually develop. The morphologic changes, time of onset, and percentage of animals affected suggest that APC is the initial manifestation of mature cataract formation in RCS rats.


COVID-19 patient transcriptomic and genomic profiling reveals comorbidity interactions with psychiatric disorders.
Psychiatric symptoms are seen in some COVID-19 patients, as direct or indirect sequelae, but it is unclear whether SARS-CoV-2 infection interacts with underlying neuronal or psychiatric susceptibilities. Such interactions might arise from COVID-19 immune responses, from infection of neurons themselves or may reflect social-psychological causes. To clarify this we sought the key gene expression pathways altered in COVID-19 also affected in bipolar disorder, post-traumatic stress disorder (PTSD) and schizophrenia, since this may identify pathways of interaction that could be treatment targets. We performed large scale comparisons of whole transcriptome data and immune factor transcript data in peripheral blood mononuclear cells (PBMC) from COVID-19 patients and patients with psychiatric disorders. We also analysed genome-wide association study (GWAS) data for symptomatic COVID-19 patients, comparing GWAS and whole-genome sequence data from patients with bipolar disorder, PTSD and schizophrenia patients. These studies revealed altered signalling and ontology pathways shared by COVID-19 patients and the three psychiatric disorders. Finally, co-expression and network analyses identified gene clusters common to the conditions. COVID-19 patients had peripheral blood immune system profiles that overlapped with those of patients with psychiatric conditions. From the pathways identified, PTSD profiles were the most highly correlated with COVID-19, perhaps consistent with stress-immune system interactions seen in PTSD. We also revealed common inflammatory pathways that may exacerbate psychiatric disorders, which may support the usage of anti-inflammatory medications in these patients. It also highlights the potential clinical application of multi-level dataset studies in difficult-to-treat psychiatric disorders in this COVID-19 pandemic.


Impact of Hepatic and Extrahepatic Insults on the Outcome of Acute-on-Chronic Liver Failure.
To study the differences in outcome and predictors of mortality in acute-on-chronic liver failure (ACLF) precipitated by hepatic or extrahepatic insults. Consecutive patients of cirrhosis with acute decompensation were prospectively included and followed up for 90 days from admission. ACLF was defined based on chronic liver failure (CLIF) acute-on-chronic liver failure in cirrhosis (CANONIC study) criteria. Acute worsening due to acute viral hepatitis A and E, hepatitis B flare, alcoholic hepatitis, autoimmune hepatitis flare, or drug-induced liver injury were categorized as hepatic ACLF and that due to bacterial infection, upper gastrointestinal bleed or surgery as extrahepatic ACLF. Patients with both hepatic and extrahepatic insults were included in combined insult group. Of 179 patients of acute decompensation, 122 had ACLF (hepatic insults 47 and extrahepatic insults 51). Alcohol (64.8%) was the most common etiology of cirrhosis while infection (36%) was the most common acute insult followed by alcoholic hepatitis (24.6%). Higher proportion of extrahepatic ACLF patients had history of prior decompensation than hepatic ACLF patients (62.7% vs. 27.7%, P < 0.001). There was no difference in mortality among hepatic and extrahepatic ACLF groups at 28 and 90 days (53.2% vs. 56.9%, P = 0.715 and 85% vs. 74.5%, P = 0.193, respectively). Area under receiver-operating curve (AUROC) for 28-day mortality in extrahepatic ACLF group was 0.788, 0.724, 0.718, 0.634, and 0.726 and in hepatic-ACLF group was 0.786, 0.625, 0.802, 0.761, and 0.648 for chronic liver failure-sequential organ failure assessment (CLIF-SOFA), model for end stage liver disease (MELD), integrated MELD score (iMELD), acute physiology and chronic health evaluation score (APACHE-II), and Child-Turcotte-Pugh score scores, respectively. There is no difference in mortality among hepatic and extrahepatic ACLF groups at 28 and 90 days. iMELD and CLIF-SOFA have highest AUROC to predict 28-day mortality in hepatic and extrahepatic ACLF groups, respectively.


Neuronal firing in the inferotemporal cortex of the monkey in a visual memory task.
The objective of this study was to elucidate the functional role of neurons in the inferotemporal cortex of the primate. Single unit activity was recorded with microelectrodes in monkeys performing a visual delayed matching-to-sample task. On each trial, the animal was exposed briefly to a color - the sample - and, after a period of delay, had to select the same color among two or four colors simultaneously presented. Thus, correct performance of the task required perception, retention, and recognition of the sample color for every trial. A large number of inferotemporal units were seen to react to the stimuli with changes of firing frequency. Many units showed color-dependent reactions, suggesting their involvement in perception and discrimination of colors. A substantial contingent of cells showed increased, sustained, and in some cases, color-dependent discharge during the delay which was not necessarily preceded or succeeded by firing changes in sample or match periods. It is proposed that those cells were engaged in temporary retention of the sample stimulus. Since most of the inferotemporal units examined showed firing changes in more than one period of a trial, they appeared to be involved in more than one of the operations required by the task. Thus, the data do not support a clear-cut topographic separation of visual functions within the inferotmeporal cortex. However, neurons that appear to participate in visual memory, either exclusively or in addition to other functions, are concentrated in the cortex of the lower banks of the superior temporal sulcus.


Diversity and evolution of B-family DNA polymerases.
B-family DNA polymerases (PolBs) represent the most common replicases. PolB enzymes that require RNA (or DNA) primed templates for DNA synthesis are found in all domains of life and many DNA viruses. Despite extensive research on PolBs, their origins and evolution remain enigmatic. Massive accumulation of new genomic and metagenomic data from diverse habitats as well as availability of new structural information prompted us to conduct a comprehensive analysis of the PolB sequences, structures, domain organizations, taxonomic distribution and co-occurrence in genomes. Based on phylogenetic analysis, we identified a new, widespread group of bacterial PolBs that are more closely related to the catalytically active N-terminal half of the eukaryotic PolEpsilon (PolEpsilonN) than to Escherichia coli Pol II. In Archaea, we characterized six new groups of PolBs. Two of them show close relationships with eukaryotic PolBs, the first one with PolEpsilonN, and the second one with PolAlpha, PolDelta and PolZeta. In addition, structure comparisons suggested common origin of the catalytically inactive C-terminal half of PolEpsilon (PolEpsilonC) and PolAlpha. Finally, in certain archaeal PolBs we discovered C-terminal Zn-binding domains closely related to those of PolAlpha and PolEpsilonC. Collectively, the obtained results allowed us to propose a scenario for the evolution of eukaryotic PolBs.


[Prediction of mortality and quality of life in polytraumatized patients: APACHE II versus APACHE III].
To evaluate the APACHE II and III prognostic assessment systems as predictors of mortality in polytraumatized patients and to compare each system's admission assessments to the patient's quality of life one month, six months and one year later. A prospective study of 130 polytraumatized patients admitted to the critical care unit was carried out. A polytraumatized patient was defined as "presenting two or more traumatic lesions that were immediately or eventually life-threatening". We studied age, sex, type of trauma, mortality, mean APACHE II and APACHE III scores during the first three days in the critical care unit for patients who survived and those who died, estimating relative risk of mortality by APACHE II and APACHE III, and quality of life one month, six months and one year after admission using Karnofsky scores (KPS). Mean patient age was 46.47 +/- 20.19 years; 78% were men and 22% women. Overall mortality was 20%. The most frequent trauma was craniocerebral (70.76%), followed by injuries ot the chest (53.8%) and extremities (46.15%). Mean APACHE II and APACHE III scores during the first three days were 10.22 +/- 5.33 and 32.75 +/- 16.42, respectively. Mean APACHE II and III scores were significantly higher (p = 0.02) in patients who died than in survivors (14.33 +/- 5.43 and 9.7 +/- 5.2, respectively, for APACHE II; and 43.27 +/- 17.68 and 30.16 +/- 15.79, respectively, for APACHE III). For each point increase of APACHE II or III scores, relative risk of mortality increased 1.09% (p = 0.03) and 1.02% (p = 0.03), respectively. Patients with APACHE II scores over 12 were 3.53 times more likely to die (p = 0.02). Patients with APACHE III scores over 35 were 3.05 times more likely to die (p = 0.02). One month after admission, 6.4% of patients had a KPS of 100 (normal, as before trauma), 35.8% achieved this score six months after admission and 82% after one year (p = 0.01). Mean APACHE III upon admission was significantly lower (p = 0.045) in patients who achieved KPS 100. The APACHE II and APACHE III prognostic systems predict relative risk of death in polytraumatized patients. APACHE III, but not APACHE II, at admission is significantly related to quality of life one year later.


Pulmonary findings in Churg-Strauss syndrome in chest X-rays and high resolution computed tomography at the time of initial diagnosis.
Churg-Strauss syndrome (CSS) is a rare, systemic, necrotizing, small- and middle-sized vessel vasculitis which is accompanied by blood eosinophilia, eosinophil infiltration of various tissues, and bronchial asthma. The lungs are the organs most often involved in CSS. The aim of this study was a retrospective evaluation of the pulmonary findings in chest X-rays and high resolution computed tomography (HRCT) in CSS patients at the time of initial diagnosis and to determine their frequency, character, and location. Seventeen CSS patients were studied (12 women; 5 men; aged 29-56 years). In all patients at the time of initial diagnosis, chest X-rays were performed, and in 15 patients, HRCT was performed additionally. The radiological images were evaluated independently by two radiologists who reached a decision by consensus. Out of 17 patients studied, chest X-rays revealed parenchymal abnormalities in 11, pleural effusion in three, and bronchial wall thickening in one. In five patients, no abnormalities in chest X-rays were found. In HRCT, abnormalities were found in all patients (15 patients, 100%). Predominant HRCT findings consisted of: ground-glass opacities and consolidations found in 13 patients (86.7%). Additionally, in four patients, pulmonary micronodules were described; in ten, interlobular septal thickening; in three, linear opacities; in ten, bronchial wall thickening and/or bronchial dilatation; and in three, pleural effusions. Ground-glass opacities and consolidation abnormalities distribution pattern were peripheral in seven and random in six patients. In patients with CSS, the most common pulmonary radiological findings are parenchymal opacities, which may be peripheral or random in distribution. Pathologic changes were found in 70.6% of patient in chest X-rays, and in 100%, when HRCT was performed. These changes are nonspecific; however, they should be not overlooked, as they may help in establishing the diagnosis and suggest the prognosis.


Does children's healthcare seeking change after participation in a musculoskeletal study? A register-based study.
Participating in research studies often involves interactions with healthcare professionals, potentially influencing the participant's future help-seeking behaviour. We investigated whether participating in the Childhood Health Activity and Motor Performance School Study - Denmark (CHAMPS) (2008-2014), which involved telephone consultations and clinical assessments by healthcare professionals with participants experiencing musculoskeletal complaints, changed frequency of contacts with primary public healthcare services among participants over the subsequent five-years-period, compared to non-participating children. Using Danish health register data from 1998 to 2020, we compared CHAMPS participant's and two control group's contacts with private physiotherapists, chiropractors (outside hospitals), and general practitioners: a random 10% sample of children from Denmark (National Controls), and a secondary local control group (Local Controls) during three periods: Before (1998-31.10.2008), during (01.11.2008-20.06.2014), and after (21.06.2014-31.12.2019) the CHAMPS-study. Separate multivariable Poisson regression models were used to assess the differences between groups for the outcome variables: contacts with physiotherapists, chiropractors, and general practitioners, and overall contacts. Compared to National Controls, the CHAMPS-Group had fewer physiotherapy contacts before the study with an estimated mean of 0.01 vs 0.02 per person-year, and after (0.13 vs 0.18 per person-year), corresponding to a crude incidence rate ratio (IRR) of 0.69 (95% confidence intervals (CI): 0.58-0.83) after the study period. However, they had more chiropractor contacts before (0.05 vs 0.03), and after (0.21 vs 0.09) the study, with a crude IRR of 2.29 (95% CI: 1.93-2.71) after the study period. General practice contacts were equal for the CHAMPS-group compared to national controls (5.84 vs 5.84) before the study but reduced during and after (3.21 vs 3.71), with a crude IRR of 0.86 (95% CI: 0.83-0.90) after the study. Comparable patterns of contacts changes from before to after the study were observed between the CHAMPS-group and the Local Controls except for physiotherapy which was equal between the two groups after the study. Our findings suggest that research studies involving systematic engagement with participants experiencing musculoskeletal complaints can influence subsequent healthcare-seeking behaviour. Future research should address the influence of health literacy, health education, and healthcare provider recommendations on healthcare decisions during such research studies.


Transcriptome analysis of endothelial cell gene expression induced by growth on matrigel matrices: identification and characterization of MAGP-2 and lumican as novel regulators of angiogenesis.
Remodeling of vascular microenvironments during normal and tumor-induced angiogenesis is an important, yet poorly understood mechanism by which endothelial cells (ECs) contribute to the activation or resolution of angiogenesis. We used microarray analyses to monitor changes in the transcriptome of ECs undergoing angiogenesis when cultured onto Matrigel matrices. This strategy identified 308 genes whose expression in ECs was altered at least 3-fold by angiogenesis, of which 63 genes were found to encode for secretory proteins. In vitro assays that modeled key steps in the angiogenic process showed that several identified genes possessed pro- or anti-angiogenic activities (e.g., SMOC-2, secreted modular calcium-binding protein-2; CRELD-2, cysteine-rich with EGF-like domains-1; MAGP-2, microfibril-associated glycoprotein-2; lumican; and ECM-1, extracellular matrix protein-1). In particular, MAGP-2 expression potentiated EC proliferation and p38 MAPK activation stimulated by the pro-angiogenic factors, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF); it also stimulated EC invasion and angiogenic sprouting, and more importantly, promoted the development and infiltration of vessels into Matrigel plugs implanted into genetically normal mice. Conversely, lumican inhibited EC activation of p38 MAPK, as well as their invasion, angiogenic sprouting, and vessel formation in mice. Collectively, our findings provide new insights into how EC stromal remodeling regulates angiogenesis activation and resolution, as well as identify two novel EC-secreted stromal proteins that modulate angiogenesis both in vitro and in vivo.


Resistance Breeding of Common Bean Shapes the Physiology of the Rhizosphere Microbiome.
The taxonomically diverse rhizosphere microbiome contributes to plant nutrition, growth and health, including protection against soil-borne pathogens. We previously showed that breeding for Fusarium-resistance in common bean changed the rhizosphere microbiome composition and functioning. Here, we assessed the impact of Fusarium-resistance breeding in common bean on microbiome physiology. Combined with metatranscriptome data, community-level physiological profiling by Biolog EcoPlate analyses revealed that the rhizosphere microbiome of the Fusarium-resistant accession was distinctly different from that of the Fusarium-susceptible accession, with higher consumption of amino acids and amines, higher metabolism of xylanase and sialidase, and higher expression of genes associated with nitrogen, phosphorus and iron metabolism. The resistome analysis indicates higher expression of soxR, which is involved in protecting bacteria against oxidative stress induced by a pathogen invasion. These results further support our hypothesis that breeding for resistance has unintentionally shaped the assembly and activity of the rhizobacterial community toward a higher abundance of specific rhizosphere competent bacterial taxa that can provide complementary protection against fungal root infections.


Disease trajectories in behavioural variant frontotemporal dementia, primary psychiatric and other neurodegenerative disorders presenting with behavioural change.
Behavioural variant frontotemporal dementia (bvFTD) is characterized by behavioural and social cognitive disturbances, while various psychiatric and neurodegenerative disorders may have similar clinical symptoms. Since neurodegenerative disorders are eventually progressive, whereas primary psychiatric disorders are not, this study aimed to investigate whether the change in clinical symptoms over time differed between groups and which biomarkers predicted rate of decline. Disease trajectories (median follow-up = 3 years) of frontal and stereotyped behaviour, general and frontal cognitive functioning, and social cognition were examined in bvFTD (n = 34), other neurodegenerative (n = 28) and primary psychiatric disorders (n = 43), all presenting with late-onset frontal lobe syndrome (45-75 years), using linear mixed models. To gain more insight in underlying pathological processes driving disease progression, we studied the association of baseline cerebrospinal fluid (CSF) (neurofilament light (NfL) and YKL-40 levels, phosphotau181 to total tau ratio) and neuroimaging markers with disease trajectories. Frontal behavioural symptoms (e.g., disinhibition, apathy) worsened over time in bvFTD, whereas they improved in psychiatric disorders and remained stable in other neurodegenerative disorders. General and frontal cognitive decline was observed in bvFTD and other neurodegenerative disorders, but not in psychiatric disorders. None of the groups showed change in stereotypy and social cognition. For all diagnostic groups, higher CSF NfL levels were associated with faster frontal cognitive decline. A modest association was observed between caudate volume and stereotyped behaviour. Tracking frontal behavioural symptoms and cognition has potential to distinguish bvFTD from other disorders. CSF NfL levels seem to be associated with decline in frontal cognitive functioning.


Memory function and the hippocampus.
There has been a long tradition in memory research of adopting the view of a vital role of the medial temporal lobe and especially the hippocampus in declarative memory. Despite the broad support for this notion, there is an ongoing debate about what computations are performed by the different substructures. The present chapter summarizes several accounts of hippocampal functions in terms of the cognitive processes subserved by these structures, the information processed, and the underlying neural operations. Firstly, the value of the distinction between recollection and familiarity for the understanding of the role the hippocampus plays in memory is discussed. Then multiple lines of evidence for the role of the hippocampus in memory are considered. Cumulating evidence suggests that the hippocampus fosters the binding of disparate cortical representations of items and their spatiotemporal context into a coherent representation by means of a sparse conjunctive neural coding. This association of item and context will then lead to the phenomenological experience of recollection. In contrast, surrounding cortical areas have broader neural coding that provide a scalar signal of the similarity between two inputs (e.g. between the encoding and the retrieval). By this they form the basis of a feeling of familiarity, but also might encode the commonalities between these different inputs. However, a more complete picture of the importance of the hippocampus for declarative memories can only be drawn when the interactions of the medial temporal lobe with other brain areas are also taken into account.


Relative effect of transcription-level and translation-level control of protein synthesis during early development of the sea urchin.
Newly synthesized proteins were pulse labeled with radioactive amino acids at several developmental stages of the sea urchin, Arbacia punctulata-in normal embryos and in embryos continuously exposed to the drug Dactinomycin. The soluble proteins were fractionated by electrophoresis through polyacrylamide gels. A simple procedure for complete solubilization of the proteins in slices of gels into a solution of toluene for liquid scintillation counting, and a computer program to display and to compare statistically the distributions of (14)C- and (3)H-labeled proteins fractionated on the same gel, allowed quantitative statements to be made on the relative changes in protein synthesis. The conclusion is that translation-level control provides most of the changes in protein synthesis that occur between fertilization and hatched blastula stage.


Depression among Muslims in the United States: Examining the Role of Discrimination and Spirituality as Risk and Protective Factors.
Depression is a widespread challenge that affects people in all cultures. Yet, despite the growth of the Muslim population in the United States, little research has been conducted on this topic with members of this cultural group. To address this gap in the literature, the present study examines the effect of discrimination and spirituality on depression with a sample of self-identified Muslims (N = 269). Consistent with our expectations, discrimination was a risk factor and spirituality was a protective factor. For instance, Muslims who reported being called offensive names were more likely (odds ratio [OR] = 3.39, 95% confidence interval [CI] = 1.82, 6.32) to report clinically significant levels of depressive symptoms compared with those who were not called offensive names, whereas saying daily prayers was associated with a lower likelihood of reporting elevated levels of symptoms (OR = 0.74, 95% CI = 0.55, 0.97). The article concludes with a discussion of the implication of the results as they intersect social work practice and of avenues for future research.


Mesonephric adenocarcinoma of the uterine corpus: a case report and review of the literature.
Mesonephric adenocarcinoma (MA) is a rare tumor of the female genital tract, mainly in the cervix and vagina, which is usually associated with mesonephric remnants or mesonephric hyperplasia. In the uterus corpus, MA is as rare as mesonephric structures, and only a few cases have been previously reported. Here, we report a rare case of MA of the uterine corpus. A 73-year-old woman presented with multiple nodules in the bilateral lung. Abdominal computed tomography scan confirmed a uterine tumor measuring 8.6 cm. All tumor markers, including CA125, were within normal limits. A total hysterectomy and bilateral salpingo-oophorectomy were performed. The tumor was confined to the myometrium and showed strong resemblance to cervical MA despite the absence of mesonephric hyperplasia or remnants. The most striking pattern consisted of large sheets of small round tubules, often with densely eosinophilic secretions in the lumen. In addition, the ductal pattern simulating endometrioid adenocarcinoma was also noted. A mixture of tubular and ductal patterns, most predominantly seen, formed more complex tubules and cribriform structures. Other elements consisted of a retiform pattern, serous adenocarcinoma-like papillary budding, and glomeruloid morphology. Immunohistochemically, the tumor cells were positive not only for cytokeratin, epithelial membrane antigen, and vimentin but also for CD10 and calretinin. No staining was identified for estrogen receptor, progesterone receptor, or androgen receptor. Adjuvant chemotherapy was begun for the patient, who is alive with disease 28 months later. We review the previously published cases of MA and discuss the principal differential diagnosis of MA in the uterine corpus.


The galanin receptor-3 antagonist, SNAP 37889, inhibits cue-induced reinstatement of alcohol-seeking and increases c-Fos expression in the nucleus accumbens shell of alcohol-preferring rats.
This study aimed to investigate the effects of the galanin-3 receptor antagonist, SNAP 37889, on c-Fos protein expression after cue-induced reinstatement of alcohol-seeking in the brains of alcohol-preferring rats. Eighteen alcohol-preferring rats were trained to self-administer 10% v/v ethanol in the presence of response-contingent cues, which was followed by extinction. Rats were then treated with SNAP 37889 (30 mg/kg, i.p.) or vehicle, before being tested for cue-induced reinstatement. Administration of SNAP 37889 reduced cue-induced reinstatement of ethanol-seeking behaviour. To examine the effect of SNAP 37889 and cue-induced reinstatement on neuronal activation, c-Fos expression was measured in subregions of the medial prefrontal cortex and nucleus accumbens. SNAP 37889 administration increased c-Fos immunoreactivity in the nucleus accumbens shell, but was without effect in the nucleus accumbens core and the medial prefrontal cortex. Dual-label Fos/tyrosine hydroxylase immunohistochemistry was used to examine the effects of SNAP 37889 on dopamine neurons in the ventral tegmental area; however, no differences between SNAP 37889 and vehicle-treated rats were found. These data support previous findings of galanin-3 receptor involvement in cue-induced reinstatement of alcohol-seeking behaviour, and provide novel evidence that the ability of galanin-3 receptor antagonism to attenuate cue-induced reinstatement relates to activation of the nucleus accumbens shell.


Nucleotide sequences of three different isoforms of beta-tubulin cDNA from Chinese hamster ovary cells.
The complete cDNA sequences of two clones encoding beta-tubulin isotypes and the partial sequence of a third isoform from Chinese hamster ovary cells have been determined. The deduced amino acid sequences of the three isoforms show extensive homology to each other as well as with other alpha and beta-tubulin sequences from various species. These results provide evidence for the expression of three different isoforms of beta-tubulin in Chinese hamster ovary cells.


Potential biomarkers in the urine of myocardial infarction rats: a metabolomic method and its application.
A metabolomic method using reversed-phase liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was developed to obtain a systematic view of the development and progression of myocardial infarction (MI). By combining with partial least squares discriminant analysis (PLS-DA), 16 biomarkers in rat urine were identified and eight of them were related to the pathway of energy metabolism. Among the regulated pathways, the citric acid cycle related network was acutely perturbed. The metabolomic results not only supplied a systematic view of the development and progression of MI but also provided the theoretical basis for the prevention or treatment of MI. The developed method was also used to analyze the therapeutic effects of a traditional Chinese medicine (TCM) named Shexiang Baoxin Pill (SBP), a widely used anti-MI medicine in clinics. The results showed that SBP administration could provide satisfactory effects on MI through partially regulating the perturbed pathway of energy metabolism.


Alpha-Linolenic and Linoleic Fatty Acids in the Vegan Diet: Do They Require Dietary Reference Intake/Adequate Intake Special Consideration?
Good sources of the long-chain n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) include cold-water fish and seafood; however, vegan diets (VGNs) do not include animal-origin foods. Typically, US omnivores obtain enough dietary EPA and DHA, but unless VGNs consume algal n-3 supplements, they rely on endogenous production of long-chain fatty acids. VGN diets have several possible concerns: (1) VGNs have high intakes of linoleic acid (LA) as compared to omnivore/non-vegetarian diets. (2) High intakes of LA competitively interfere with the endogenous conversion of alpha-linolenic acid (ALA) to EPA and DHA. (3) High somatic levels of LA/low ALA indicate a decreased ALA conversion to EPA and DHA. (4) Some, not all VGNs meet the Dietary Reference Intake Adequate Intake (DRI-AI) for dietary ALA and (5) VGN diets are high in fiber, which possibly interferes with fat absorption. Consequently, health professionals and Registered Dietitians/Registered Dietitian Nutritionists working with VGNs need specific essential fatty acid diet guidelines. The purpose of this review was: (1) to suggest that VGNs have a DRI-AI Special Consideration requirement for ALA and LA based on VGN dietary and biochemical indicators of status and (2) to provide suggestions to ensure that VGNs receive adequate intakes of LA and ALA.


The impact of TennCare on patient satisfaction with care.
To measure the level of satisfaction with care by Medicaid-eligible patients before and after implementation of a mandatory managed care plan known as TennCare. We used multivariate logit analysis of survey data to calculate the effects of TennCare on patient satisfaction for TennCare patients compared to those on traditional Medicaid, using North Carolina as a control state. Patients were respondents to a survey conducted in late 1996 and early 1997 who had been admitted to hospitals in 1993 and 1995 for labor/delivery (n = 986), acute myocardial infarction (n = 457), and head trauma (n = 248). Dependent variables were yes/no responses to satisfaction questions for labor/delivery and 5-category ordered responses for adults. We found no statistically significant differences in satisfaction between TennCare and traditional Medicaid for either pediatric or adult hospital patients. Generally, TennCare recipients had satisfaction levels as good or better than traditional Medicaid recipients. For pediatric care, TennCare odds ratios ranged from 1.00 to 2.17, the latter for satisfaction with care received (P = 0.107). For adult care, odds ratios ranged from 0.77 to 1.23, the latter for satisfaction with cost of care (P = 0.547). For many dimensions of care, lower rates of satisfaction were reported for respondents who were uninsured, less educated, and in poor health. For adult care, blacks or Hispanics tended to be less satisfied with some aspects of care. TennCare did not reduce patient satisfaction with care among those who were hospitalized.


Apolipoprotein E mimetic peptide CN-105 improves outcome in a murine model of SAH.
Subarachnoid haemorrhage (SAH) accounts for 3% of all strokes, and is associated with significant morbidity and mortality. There is growing evidence implicating apolipoprotein E (apoE) in mediating adaptive anti-inflammatory and neuroprotective responses following ischaemic and traumatic brain injury. In the current study, we test the efficacy of a small apoE mimetic peptide, CN-105 in a murine model of SAH. Mice subjected to SAH received repeated intravenous injections of CN-105 every 12 hours for 3 days, with the first dose given 2 hours after injury. Daily functional outcomes were assessed by rotarod and neurological severity score. Haemorrhage grade and cerebral vascular diameters were measured at 5 days post-SAH. Cerebral microgliosis, neuronal degeneration and survival were analysed at 5 and 35 days post-SAH, respectively. CN-105 reduces histological evidence of inflammation, reduces vasospasm and neuronal injury and is associated with improved long-term behavioural outcomes in a murine model of SAH. Given its favourable pharmacokinetic profile, central nervous system penetration and demonstration of clinical safety, CN-105 represents an attractive therapeutic candidate for treatment of brain injury associated with SAH.


Cell replacement and regeneration therapy for diabetes.
Reduction of beta cell function and a beta cell mass is observed in both type 1 and type 2 diabetes. Therefore, restoration of this deficiency might be a therapeutic option for treatment of diabetes. Islet transplantation has benefits, such as reduced incidence of hypoglycemia and achievement of insulin independence. However, the major drawback is an insufficient supply of islet donors. Transplantation of cells differentiated in vitro or in vivo regeneration of insulin-producing cells are possible approaches for beta cell/islet regenerative therapy. Embryonic and adult stem cells, pancreatic ductal progenitor cells, acinar cells, and other endocrine cells have been shown to differentiate into pancreatic beta cells. Formation of fully functional beta cells and the safety of these cells are critical issues for successful clinical application.


Ongoing Effects of SARS-CoV-2 Infection on Arterial Stiffness in Healthy Adults.
There are limited data on the arterial stiffness changes after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The present study evaluated the changes in arterial stiffness in completely healthy patients who had a SARS-CoV-2 infection using the cardio-ankle vascular index (CAVI). The study included 70 patients with SARS-CoV-2 infection between December 2020 and June 2021. A cardiac evaluation was carried out, including chest X-ray, electrocardiography (ECG), and echocardiography in all patients. Within the 1st month and 7th month CAVI was measured. Mean age was 37.8 ± 10.0 years, and 41/70 were female. Mean height was 168.6 ± 9.5 cm, mean weight 73.2 ± 15.1 kg, and mean body mass index (BMI) of the group was 25.6 ± 4.2, respectively. CAVI results from the right arm were 6.45 ± .95 at 1-month follow-up and 6.68 ± 1.05 at 7 months follow-up (P = .016) and from the left arm were 6.43 ± 1.0 at 1-month follow-up, 6.70 ± 1.05 at 7-month follow-up (P = .005). Our results showed an ongoing injury in the arterial system after healthy SARS-COV-2 patients during 7 months, as represented by CAVI measurements.


Antibiotic Resistance and the Biology of History.
Beginning in the 1940s, mass production of antibiotics involved the industrial-scale growth of microorganisms to harvest their metabolic products. Unfortunately, the use of antibiotics selects for resistance at answering scale. The turn to the study of antibiotic resistance in microbiology and medicine is examined, focusing on the realization that individual therapies targeted at single pathogens in individual bodies are environmental events affecting bacterial evolution far beyond bodies. In turning to biological manifestations of antibiotic use, sciences fathom material outcomes of their own previous concepts. Archival work with stored soil and clinical samples produces a record described here as 'the biology of history': the physical registration of human history in bacterial life. This account thus foregrounds the importance of understanding both the materiality of history and the historicity of matter in theories and concepts of life today.


HC-toxin.
HC-toxin is a cyclic tetrapeptide of structure cyclo(D-Pro-L-Ala-D-Ala-L-Aeo), where Aeo stands for 2-amino-9,10-epoxi-8-oxodecanoic acid. It is a determinant of specificity and virulence in the interaction between the producing fungus, Cochliobolus carbonum, and its host, maize. HC-toxin qualifies as one of the few microbial secondary metabolites whose ecological function in nature is understood. Reaction to C. carbonum and to HC-toxin is controlled in maize by the Hm1 and Hm2 loci. These loci encode HC-toxin reductase, which detoxifies HC-toxin by reducing the 8-carbonyl group of Aeo. HC-toxin is an inhibitor of histone deacetylases (HDACs) in many organisms, including plants, insects, and mammals, but why inhibition of HDACs during infection by C. carbonum leads to disease is not understood. The genes for HC-toxin biosynthesis (collectively known as the TOX2 locus) are loosely clustered over >500 kb in C. carbonum. All of the known TOX2 genes are present in multiple, functional copies and are absent from natural toxin non-producing isolates. The central enzyme in HC-toxin biosynthesis is a 570-kDa non-ribosomal synthetase encoded by a 15.7-kb open reading frame. Other genes known to be required for HC-toxin encode alpha and beta subunits of fatty acid synthase, which are presumed to contribute to the synthesis of Aeo; a pathway-specific transcription factor; an efflux carrier; a predicted branched-chain amino acid aminotransferase; and an alanine racemase.


Genomes of replicatively senescent cells undergo global epigenetic changes leading to gene silencing and activation of transposable elements.
Replicative cellular senescence is an important tumor suppression mechanism and also contributes to aging. Progression of both cancer and aging include significant epigenetic components, but the chromatin changes that take place during cellular senescence are not known. We used formaldehyde assisted isolation of regulatory elements (FAIRE) to map genome-wide chromatin conformations. In contrast to growing cells, whose genomes are rich with features of both open and closed chromatin, FAIRE profiles of senescent cells are significantly smoothened. This is due to FAIRE signal loss in promoters and enhancers of active genes, and FAIRE signal gain in heterochromatic gene-poor regions. Chromatin of major retrotransposon classes, Alu, SVA and L1, becomes relatively more open in senescent cells, affecting most strongly the evolutionarily recent elements, and leads to an increase in their transcription and ultimately transposition. Constitutive heterochromatin in centromeric and peri-centromeric regions also becomes relatively more open, and the transcription of satellite sequences increases. The peripheral heterochromatic compartment (PHC) becomes less prominent, and centromere structure becomes notably enlarged. These epigenetic changes progress slowly after the onset of senescence, with some, such as mobilization of retrotransposable elements becoming prominent only at late times. Many of these changes have also been noted in cancer cells.


Control of parasitemia and survival during Trypanosoma brucei brucei infection is related to strain-dependent ability to produce IL-4.
We studied non-MHC gene-dependent expression of a number of cytokines in relation to host defense and survival during Trypanosoma brucei brucei (Tbb) infection in mice. In particular, the role of IL-4 was explored with use of genomically IL-4-disrupted mice and in vivo Ab blocking. Splenocytes from MHC-identical B10.Q (relatively resistant) mice showed day 5 postinfection higher numbers of IL-4 mRNA expressing cells than C3H.Q (highly susceptible). A trypanosome-derived lymphocyte triggering factor, which is released by Tbb to polyclonally activate CD8+ T cells, stimulated naive splenocytes in vitro to a higher IL-4 response in B10.Q than in C3H.Q mice. The C3H.Q mice developed an extremely high parasitemia, showed a low Ab response against the variant surface glycoprotein (VSG), and had a mean survival time of 42 days. Conversely, B10.Q mice had lower parasitemia, mounted higher anti-VSG response, and had a mean survival time of 56 days. Deletion of the IL-4 gene had no influence on the infection in C3H.Q mice, while in B10.Q mice the deletion was associated with lower anti-VSG Ab levels and higher parasitemia. Paradoxically, B10.Q mice with disrupted IL-4 gene survived longer than the wild type. Anti-IL-4 Ab-blocking experiments in vivo displayed an enhanced parasitemia and prolonged survival in infected B10.Q mice. We conclude that 1) a non-MHC gene-related and CD8+-dependent ability to produce IL-4 partly determines the susceptibility to Tbb infection; and 2) IL-4, although involved in controlling the levels of parasitemia by its effects on immunoglobulin synthesis, also can have toxic effects on the animals.


Efficacy of fidaxomicin versus vancomycin as therapy for Clostridium difficile infection in individuals taking concomitant antibiotics for other concurrent infections.
Treatment guidelines recommend stopping all implicated antibiotics at the onset of Clostridium difficile infection (CDI), but many individuals have persistent or new infections necessitating the use of concomitant antibiotics (CAs). We used data from 2 phase 3 trials to study effects of CAs on response to fidaxomicin or vancomycin. Subjects with CDI were treated for 10 days with fidaxomicin 200 mg every 12 hours or vancomycin 125 mg every 6 hours, assessed for resolution of symptoms, and followed up for an additional 4 weeks for evidence of recurrence. Rates of cure, recurrence, and global cure (cure without recurrence) were determined for subgroups of subjects defined by CA use and treatment group. CAs were prescribed for 27.5% of subjects during study participation. The use of CAs concurrent with CDI treatment was associated with a lower cure rate (84.4% vs 92.6%; P < .001) and an extended time to resolution of diarrhea (97 vs 54 hours; P < .001). CA use during the follow-up was associated with more recurrences (24.8% vs 17.7%; not significant), and CA administration at any time was associated with a lower global cure rate (65.8% vs 74.7%; P = .005). When subjects received CAs concurrent with CDI treatment, the cure rate was 90.0% for fidaxomicin and 79.4% for vancomycin (P = .04). In subjects receiving CAs during treatment and/or follow-up, treatment with fidaxomicin compared with vancomycin was associated with 12.3% fewer recurrences (16.9% vs 29.2%; P = .048). Treatment with CAs compromised initial response to CDI therapy and durability of response. Fidaxomicin was significantly more effective than vancomycin in achieving clinical cure in the presence of CA therapy and in preventing recurrence regardless of CA use.


Measuring variability in phytotoxicity testing using crop and wild plant species.
A series of experiments was conducted to assess the level of variability in phytotoxicity testing and to investigate factors that may explain some of the observed uncertainties and inconsistencies. The work was conducted in greenhouse or growth chamber environments with plants growing individually in pots and harvested 28 d after spraying with two herbicides, glyphosate and atrazine, as formulated products. Between six and 10 doses were used on five or six replicates, necessitating over 4,500 individually growing plants. In the first set of experiments, several ecotypes (originating from different areas of the world) of eight wild plant species were tested. Significant differences in sensitivity to atrazine and glyphosate were found among ecotypes of most species tested. In the second suite of experiments, the reproducibility of results during different seasons (when growing conditions vary) was investigated using three crops and four wild plant species. Results showed that seasonal variability elicited a pronounced discrepancy in response between plants tested at different times of the year. It was found that no consistent effects could be attributed to the biotic or abiotic factors investigated. Several ecotypes of the same species differed in their seed size, percentage germination, or germination requirements, as well as in growth patterns, but these differences could not explain differences in herbicide sensitivity. Likewise, differences in phytotoxicity could not be attributed to factors such as temperature, light intensity, and sunlight duration. The present study supports the inclusion of an uncertainty factor in risk assessments to account for the intrinsic variability in plant sensitivity to herbicides.


Renoprotective Effects of Aldose Reductase Inhibitor Epalrestat against High Glucose-Induced Cellular Injury.
Diabetic nephropathy (DN) is the leading cause of end stage renal disease worldwide. Increased glucose flux into the aldose reductase (AR) pathway during diabetes was reported to exert deleterious effects on the kidney. The objective of this study was to investigate the renoprotective effects of AR inhibition in high glucose milieu in vitro. Rat renal tubular (NRK-52E) cells were exposed to high glucose (30 mM) or normal glucose (5 mM) media for 24 to 48 hours with or without the AR inhibitor epalrestat (1 μM) and assessed for changes in Akt and ERK1/2 signaling, AR expression (using western blotting), and alterations in mitochondrial membrane potential (using JC-1 staining), cell viability (using MTT assay), and cell cycle. Exposure of NRK-52E cells to high glucose media caused acute activation of Akt and ERK pathways and depolarization of mitochondrial membrane at 24 hours. Prolonged high glucose exposure (for 48 hours) induced AR expression and G1 cell cycle arrest and decreased cell viability (84% compared to control) in NRK-52E cells. Coincubation of cells with epalrestat prevented the signaling changes and renal cell injury induced by high glucose. Thus, AR inhibition represents a potential therapeutic strategy to prevent DN.


Ca2+ signal transduction related to neutral lipid synthesis in an oil-producing green alga Chlorella sp. C2.
Changes in the cytosolic Ca(2+) levels and the role of Ca(2+) signal transduction in neutral lipid synthesis in Chlorella sp. C2 under nitrogen starvation conditions were investigated. The results detected by using the scanning ion-selective electrode technique demonstrate that nitrogen starvation induced significant Ca(2+) influx across the plasma membrane into cells. Ca(2+) fluorescence imaging and flow cytometry were used to estimate the effect of this Ca(2+) influx on the generation of the Ca(2+) signal, and the results showed that the cytosolic Ca(2+) concentration increased transiently and then remained at a stable, high level when the cells were exposed to nitrogen starvation. However, the increase could be inhibited by pre-treatment with the Ca(2+) channel blockers ruthenium red, verapamil and GdCl3, indicating that both the influx of Ca(2+) from the extracellular space via Ca(2+) channels that are localized in the plasma membrane and the release of Ca(2+) from intracellular calcium storage via the internal calcium store were required for the generation and transduction of the Ca(2+) signal. During nitrogen starvation, neutral lipid synthesis in Chlorella sp. C2 in response to stress conditions was also inhibited to differing degrees by pre-treatment with the three Ca(2+) channel blockers, demonstrating the regulation of Ca(2+) via these Ca(2+) channels in neutral lipid synthesis. The results suggested that by transduction of extracellular stress signals into the cell and the regulation of the Ca(2+) signal in neutral lipid synthesis, Ca(2+) signal transduction played important roles in the response mechanism of Chlorella sp. C2 to nitrogen starvation.


Prenatal Particulate Air Pollution and Asthma Onset in Urban Children. Identifying Sensitive Windows and Sex Differences.
The influence of particulate air pollution on respiratory health starts in utero. Fetal lung growth and structural development occurs in stages; thus, effects on postnatal respiratory disorders may differ based on timing of exposure. We implemented an innovative method to identify sensitive windows for effects of prenatal exposure to particulate matter with a diameter less than or equal to 2.5 μm (PM2.5) on children's asthma development in an urban pregnancy cohort. Analyses included 736 full-term (≥37 wk) children. Each mother's daily PM2.5 exposure was estimated over gestation using a validated satellite-based spatiotemporal resolved model. Using distributed lag models, we examined associations between weekly averaged PM2.5 levels over pregnancy and physician-diagnosed asthma in children by age 6 years. Effect modification by sex was also examined. Most mothers were ethnic minorities (54% Hispanic, 30% black), had 12 or fewer years of education (66%), and did not smoke in pregnancy (80%). In the sample as a whole, distributed lag models adjusting for child age, sex, and maternal factors (education, race and ethnicity, smoking, stress, atopy, prepregnancy obesity) showed that increased PM2.5 exposure levels at 16-25 weeks gestation were significantly associated with early childhood asthma development. An interaction between PM2.5 and sex was significant (P = 0.01) with sex-stratified analyses showing that the association exists only for boys. Higher prenatal PM2.5 exposure at midgestation was associated with asthma development by age 6 years in boys. Methods to better characterize vulnerable windows may provide insight into underlying mechanisms.


Immunobiology of Newcastle Disease Virus and Its Use for Prophylactic Vaccination in Poultry and as Adjuvant for Therapeutic Vaccination in Cancer Patients.
Newcastle disease (ND) is one of the most important diseases of poultry worldwide. In the last decades, molecular research has gained a lot of new information about its causative agent, newcastledisease virus (NDV). In poultry industry, certain strains of NDV have been used for preventive vaccination for more than 60 years. NDV has also been applied to cancer patients with beneficial effects for about 50 years, but this is less well known. The molecular basis for these differential effects of NDV in birds and man have been elucidated in the last decades and are explained in this review. The anti-neoplastic and immune-stimulatory properties in non-permissive hosts such as mouse and man have to do with the strong type I interferon responses induced in these foreign species. Additionally, NDV has the potential to break various types of tumor resistances and also to affect liver fibrosis. A main section is devoted to the benefits of clinical application of NDV and NDV-based vaccines to cancer patients. Reverse genetics technology allowed developing NDV into a vector suitable for gene therapy. Examples will be provided in which genetically engineered NDV is being used successfully as vector against new emerging viruses.