Influence of Temperature and Humidity on the Viability of Ophiognomonia clavigignenti-juglandacearum Conidia.
Butternut canker, caused by the fungus Ophiognomonia clavigignenti-juglandacearum, primarily kills butternut (Juglans cinerea). Rain splash and local air currents are the primary means of conidia dispersal but that does not explain its long-distance spread and infection of isolated trees. Dispersal by insect or animal vectors or plant material likely necessitates the ability for conidia to tolerate drying for a period of time over variable temperature and humidity conditions. The objective of this study was to determine the influence of temperature and humidity on conidial germination and survival of air-dried conidia. Conidia collected from 1-month-old cultures germinated on water agar over a wide range of temperatures (4 to 32°C) and were viable after brief periods at 36°C when returned to a lower temperature. Viability of air-dried conidia held on nylon membranes at various temperatures and humidities varied from less than a day at 28°C and 90% relative humidity (RH) to a mean of 15 days at 20°C and 80% RH. RH had the least effect on viability at 12°C, with conidia remaining viable for 7 days at most humidity levels tested. Conidia held at 100% RH began germinating on the membranes after 21 days. Conidia in a water suspension remained viable for 168 days at all temperatures tested. These results suggest that O. clavigignenti-juglandacearum conidia may remain viable on the surface of a vector or plant material and seed for over 2 weeks, given the proper conditions, or for much longer if in water or in an environment of saturated humidity. This potential may, in part, explain the frequent presence of the disease on isolated trees.


Effect of Conjugated Linoleic Acid Intake on Endurance Exercise Performance and Anti-fatigue in Student Athletes.
Conjugated linoleic acids (CLA) intake has been reported to reduce body fat mass or increase lean body mass and to improve exercise outcome by modulating testosterone in humans. These reports have studied mostly overweight subjects; few were athletes. Therefore, in this study, the effect of CLA intake on endurance performance and anti-fatigue in student athletes was investigated. A double-blind, crossover study was conducted with 10 male student athletes. Each subject was administered with either CLA (net 0.9 g/day) or a placebo for 14 days. They were subjected to an exercise tolerance test (steady loading) using a cycle ergometer on days 0 and 14. Peak VO2 was determined for each subject using a graded loading test. The steady loading test was performed with a pedaling exercise load of 50% peak VO2 for 40 min and then with a load of 70% peak VO2 until exhaustion. Blood sampling and measurement of critical flicker frequency (CFF) were performed before and after exercise. The rate of perceived exertion (RPE) was measured serially during exercise. In the results, amount of body weight variation significantly increased and amount of body fat percentage variation tended to decrease by CLA intake, it might have an effect by increase in muscle mass. In addition, amount of exercise time variation significantly increased, amount of variation of CFF before and after exercise tended to increase, that of RPE during exercise tended to decrease, and that of creatine phosphokinase before and after exercise tended to decrease in the CLA group. These results suggested that CLA intake for 14 days might have an effect on endurance performance and anti-fatigue in student athletes.


Comparison of clinical judgment, Doppler ultrasound, and fluorescein fluorescence as methods for predicting intestinal viability in the pony.
Strangulation obstruction was induced in anesthetized ponies for periods of 2 and 3 hours by clamping 45-cm segments of jejunum and associated veins (venous strangulation obstruction) and arteries and veins (arterial and venous strangulation obstruction). Four segments were studied in each of 7 ponies allowed to survive 12 hours, 2 segments in a pony that was allowed to survive 1 hour, and 1 segment in each of 10 ponies allowed to survive 42 days after the strangulation periods ended. Fifteen minutes after the periods of strangulation obstruction ended, the viability of test segments was assessed by clinical judgment (40 segments), fluorescein fluorescence (40 segments), and Doppler ultrasound (32 segments). Because the test segments were normal at necropsy in long-term survivors, all segments were designated as viable. The overall accuracy of the methods used to predict viability was 88% for Doppler ultrasound and 53% each for clinical judgment and fluorescein fluorescence (P less than 0.005). Failures in the last 2 techniques could be attributed to their tendency to score venous strangulation obstruction segments as nonviable (90% for each). Doppler ultrasound was 94% accurate in these segments.


Mathematical anxiety is linked to reduced cognitive reflection: a potential road from discomfort in the mathematics classroom to susceptibility to biases.
When asked to solve mathematical problems, some people experience anxiety and threat, which can lead to impaired mathematical performance (Curr Dir Psychol Sci 11:181-185, 2002). The present studies investigated the link between mathematical anxiety and performance on the cognitive reflection test (CRT; J Econ Perspect 19:25-42, 2005). The CRT is a measure of a person's ability to resist intuitive response tendencies, and it correlates strongly with important real-life outcomes, such as time preferences, risk-taking, and rational thinking. In Experiments 1 and 2 the relationships between maths anxiety, mathematical knowledge/mathematical achievement, test anxiety and cognitive reflection were analysed using mediation analyses. Experiment 3 included a manipulation of working memory load. The effects of anxiety and working memory load were analysed using ANOVAs. Our experiments with university students (Experiments 1 and 3) and secondary school students (Experiment 2) demonstrated that mathematical anxiety was a significant predictor of cognitive reflection, even after controlling for the effects of general mathematical knowledge (in Experiment 1), school mathematical achievement (in Experiment 2) and test anxiety (in Experiments 1-3). Furthermore, Experiment 3 showed that mathematical anxiety and burdening working memory resources with a secondary task had similar effects on cognitive reflection. Given earlier findings that showed a close link between cognitive reflection, unbiased decisions and rationality, our results suggest that mathematical anxiety might be negatively related to individuals' ability to make advantageous choices and good decisions.


Low-Income Ethnically Diverse Children's Engagement as a Predictor of School Readiness Above Preschool Classroom Quality.
This study examined whether children's observed individual engagement with teachers, peers, and tasks related to their school readiness after controlling for observed preschool classroom quality and children's baseline skills. The sample included 211 predominately low-income, racially/ethnically diverse 4-year-old children in 49 preschool classrooms in one medium-sized U.S. city. Results indicated that children's positive engagement with (a) teachers related to improved literacy skills; (b) peers related to improved language and self-regulatory skills; and (c) tasks related to closer relationships with teachers. Children's negative engagement was associated with lower language, literacy, and self-regulatory skills, and more conflict and closeness with teachers. Effect sizes were small to medium in magnitude, and some expected relations between positive engagement and school readiness were not found.


Metabolomics technology and bioinformatics.
Metabolomics is the global analysis of all or a large number of cellular metabolites. Like other functional genomics research, metabolomics generates large amounts of data. Handling, processing and analysis of this data is a clear challenge and requires specialized mathematical, statistical and bioinformatics tools. Metabolomics needs for bioinformatics span through data and information management, raw analytical data processing, metabolomics standards and ontology, statistical analysis and data mining, data integration and mathematical modelling of metabolic networks within a framework of systems biology. The major approaches in metabolomics, along with the modern analytical tools used for data generation, are reviewed in the context of these specific bioinformatics needs.


The spindle midzone microtubule-associated proteins Ase1p and Cin8p affect the number and orientation of astral microtubules in Saccharomyces cerevisiae.
The nucleus of the budding yeast S. cerevisiae has to move to the bud neck during mitosis in order for proper DNA segregation to take place. This movement is mediated by spindle and astral microtubules, and it relies on forces generated by microtubule-associated motor proteins. When budding yeast cells express the non-cleavable cohesin subunit, Scc1-RRDD, sister chromatid separation is blocked, preventing the spindle from elongating. Thus, in the presence of Scc1-RRDD nuclear positioning is mediated solely by forces acting through astral microtubules. We have previously shown that under these conditions cells exit mitosis with the nucleus in the mother cells, and that the position of the nucleus is determined, at least in part, by the FEAR pathway, which regulates various aspects of mitotic exit. When the FEAR pathway is inactivated, cells expressing Scc1-RRDD exit mitosis with the nucleus in the daughter cells (referred to as a "daughterly phenotype"). In order to find additional proteins that participate in nuclear positioning, we screened a series of mutant strains for those that displayed a daughterly phenotype when Scc1-RRDD was expressed. The most prominent defects were seen in ase1delta and cin8delta mutant cells. Both Ase1p and Cin8p were previously shown to be nuclear and to be involved in spindle function. We show here that deletion of ASE1 or CIN8 causes a defect in SPB separation and leads to an abnormal number of astral microtubules and a change in their orientation within the cell. Taken together, these results suggest that in budding yeast Ase1p and Cin8p affect nuclear positioning through astral micro-tubule-dependent mechanisms.


Indigenous people in Aotearoa New Zealand are overrepresented in cannabis convictions.
Previous work has demonstrated that cannabis laws have had a disproportionate impact on Māori, the Indigenous people of Aotearoa New Zealand. In 2019, the New Zealand Government amended cannabis laws, providing police with the power to determine whether a therapeutic or health-centred approach would be more beneficial than a conviction. In the current study, we use population level data to assess whether this law change has ameliorated the bias in cannabis convictions for Māori. Data were drawn from the Integrated Data Infrastructure (IDI), a large government database hosted by Aotearoa New Zealand's national statistics office. In the IDI, we selected individuals who (1) were between 18 and 65, (2) were Māori or Pākehā (New Zealanders of European descent) and, (3) had any cannabis charges that proceeded to the courts. Māori ethnicity was a significant predictor of the odds of receiving a cannabis conviction for Māori males (Odds: 1.56), with a marginally significant effect for Māori females (Odds: 1.57). Further, for Māori, there was no reduction in the number of cannabis charges before vs. after the amendment to cannabis laws. The current study demonstrates that the bias in cannabis convictions for Māori remain. Given this, the New Zealand Government must follow other countries around the world and move forward on cannabis law reform.


Stage-specific expression and cellular localization of the heat shock factor 2 isoforms in the rat seminiferous epithelium.
Heat shock transcription factors (HSFs) are generally known as regulators of cellular stress response. The mammalian HSF1 functions as a classical stress factor, whereas HSF2 is active during certain developmental processes, including embryogenesis and spermatogenesis. In the present study, we examined HSF2 expression at specific stages of the rat seminiferous epithelial cycle. We found that expression of the alternatively spliced HSF2-alpha and HSF2-beta isoforms is developmentally regulated in a stage-specific manner. Studies on cellular localization demonstrated that HSF2 is present in the nuclei of early pachytene spermatocytes at stages I-IV and in the nuclei of round spermatids at stages V-VIIab. In contrast a strong HSF2 immunoreactivity was detected in small distinct cytoplasmic regions from zygotene spermatocytes to maturation phase spermatids. Immunoelectron microscopic analysis revealed that these structures are mainly cytoplasmic bridges between germ cells. Our results on cellular localization of HSF2 and stage-specific expression of the HSF2 isoforms indicate that HSF2, in addition to its function as a nuclear transcription factor, may be involved in other cellular processes during spermatogenesis, possibly in the sharing process of gene products between the germ cells.


Freestanding χ3-borophene nanoribbons: a density functional theory investigation.
Experimental observation of borophene nanoribbons (BNRs) motivated us to carry out a comprehensive investigation on BNRs, decomposed from a χ3 sheet, using density functional theory. Our results show that the stability and also the electrical and magnetic properties of the ribbons are strongly dependent on the edge configurations. We have studied two categories of ribbon: XBNRs and YBNRs. The first one is a nonmagnetic metal with armchair shaped edges, while YBNRs can be magnetic or nonmagnetic depending on the edge shape. YBNRs have four different edge types and we show that two of them are magnetic (a- and b-type edges) while the other two are nonmagnetic (c- and d-type edges). There are 10 distinct configurations possible by arranging the different edges of YBNRs. 10 percent of YBNRs are polarized asymmetrically at the edges, leading to the loss of degeneracy of the spin-up and spin-down bands in the antiferromagnetic configuration. 40 percent of YBNRs have one magnetic edge and can be promising candidates for spintronic applications due to the separation of the spin in the real space in addition to the energy space. Electronic transmission properties of the ribbons were also studied and we found that transmission channels are suppressed at the edges of XBNRs due to electron localization.


Chimeric antigen receptor T-cell immunotherapies adverse events reported to FAERS database: focus on cytopenias.
Chimeric antigen receptor (CAR) T-cell therapy presents a promising treatment for hematologic malignancies, displaying high efficacy but not being exempt from toxicity. In this observational study, we assessed adverse events (AEs) reported to the Food and Drug Adverse Event Reporting System (FAERS) including any of the six approved CAR T-cell therapies. A total of 5249 reports mentioning a CAR T-cell as a suspect product were retrieved from the FAERS database, containing a total of 24333 AEs, of which 3236 (13.3%) were cytopenias. The highest number of AEs mentioned by the report was observed for tisagenlecleucel (mean = 6.7), with the lowest for ciltacabtagene (mean = 1.3). Among all reports, hematopoietic leukopenia was the most frequently reported AEs (n = 1386, 5.7%), with hematopoietic erytropenia the least reported (n = 291, 1.2%). Tisagenlecleucel showed a high reporting odds ratio for hematopoietic erythropenia (27.28, 95%CI 14.04-53.00), leukopenia (4.04, 95%CI 3.52-4.64), and thrombocytopenia (4.01, 95%CI 3.19-5.03). Cytopenias represent one of the most frequently reported AEs in FAERS, a CAR T-cell therapy is indicated, with haematopoetic leukopenia being the most common. When comparing different CAR-T cell therapies, the cytopenias' reporting odds ratio was particularly high for tisagenlecleucel, especially in relation to hematopoietic erythropenia.


Endoscopic laser vs open approach for cricopharyngeal myotomy.
To illustrate the safety and efficacy of the endoscopic laser approach for cricopharyngeal myotomy (CPM) compared to the traditional transcervical approach. Retrospective chart review of 22 patients undergoing CPM from 1996 to 2003 at the Mayo Clinic, Jacksonville. The laser CPM technique was used in 14 patients, and an open approach in 8. The mean hospital stay and operative times were shorter for the laser group. Functional outcome analyses showed improvement in both groups. There were no major complications in the laser group, while 1 patient in the transcervical group had a pharyngocutaneous fistula. The laser technique is at least as effective as the transcervical approach for CPM to improve dysphagia symptoms in the properly selected patient, with a low risk of major complications. In this report, we provide the reader with data to support the safety and efficacy of laser CPM. B-3b.


Supporting Patient-Centered Pregnancy Counseling in Nephrology Care: A Semistructured Interview Study of Patients and Nephrologists.
Individuals with chronic kidney disease (CKD) are at increased risk of adverse pregnancy outcomes and are susceptible to disempowerment and decisional burden when receiving reproductive counseling and considering pregnancy. Nephrologists do not frequently counsel about reproductive health, and no tools exist to support patient-centered reproductive counseling for those with CKD. A total of 30 patients aged 18 to 45 years with CKD stages 1 to 5 who were assigned female sex at birth and 12 nephrologists from a single academic medical center participated in semistructured qualitative interviews. They were asked about information needs, decision support needs, and facilitators and barriers to reproductive health care and counseling. Thematic analysis was performed. The following 4 main themes were identified: (i) assessing reproductive intentions; (ii) information about reproductive health and kidney disease; (iii) reproductive risk; and (iv) communication and decision-making needs. Patients' reproductive intentions varied over time and shaped the content of information needed from nephrologists. Patients and nephrologists both felt that risk communication could be improved but focused on different aspects to improve the quality of this counseling; nephrologists focused on providing individualized risk estimates and patients focused on balancing risks with benefits and management. Patients desired nephrologists to bring up the topic of reproductive health and counseling in kidney clinic, and this is not frequently or systematically done currently. This work highlights a critical need for more dialog about reproductive health in kidney care, identified differences in what patients and nephrologists think is important in communication and decision-making, and provides an important step in developing patient-centered reproductive counseling tools in nephrology.


Real time tissue elasticity imaging using the combined autocorrelation method.
The elastic properties of tissues are expected to provide novel information for use in diagnosing pathologic changes in tissues and discriminating between malignant and benign tumors. Because it is hard to directly estimate the elastic modulus distribution from echo signals, methods for imaging the distribution of tissue strain under static compression are being widely investigated. Imaging the distribution of strain has proven to be useful for detecting disease tissues on the basis of their differences in elastic properties, although it is more qualitative than elastic modulus distribution. Many approaches to obtaining strain images from echo signals have been proposed. Most of these approaches use the spatial correlation technique, a method of detecting tissue displacement that provides maximum correlation between the echo signal obtained before and the one obtained after compression. Those methods are not suited for real-time processing, however, because of the amount of computation time they require. An alternative approach is a phase-tracking method, which is analogous to Doppler blood flowmetry. Although it can realize the rapid detection of displacement, the aliasing effect prevents its application to the large displacements that are necessary to improve the S/N ratio of the strain image. We therefore developed a more useful technique for imaging tissue elasticity. This approach, which we call the combined autocorrelation (CA) method, has the advantages of producing strain images of high quality with real-time processing and being applicable to large displacements.Numeric simulation and phantom experimentation have demonstrated that this method's capability to reconstruct images of tissue strain distribution under practical conditions is superior to that of the conventional spatial correlation method. In simulation and phantom experimentation, moreover, the image of elastic modulus distribution was also obtained by estimating stress distribution using a three-dimensional tissue model. When the proposed CA method was used to measure breast tumor specimens, the obtained strain images clearly revealed harder tumor lesions that were only vaguely resolved in B-mode images. Moreover, the results indicated the possibility of extracting the pathological characteristics of a tumor, making it useful for determining tumor type. These advantages justify the clinical use of the CA method.


Zeugmatin is part of the Z-band targeting region of titin.
Originally, zeugmatin was identified as a 600-800 kD muscle specific protein in Z-bands of cardiac and skeletal muscles by Maher et al. (1985). In this presentation we review our work on myofibrillogenesis and present evidence that zeugmatin is actually part of the Z-Band region of titin and that this region of titin plays an important role in the assembly of the Z-bands and myofibrils. Rhee et al. (1994) reported that during myofibrillogenesis, zeugmatin antibody localization is detected in fully formed Z-bands in the mature myofibrils, in the Z-bodies of the nascent myofibrils, but not in the Z-bodies of the premyofibrils. These observations lead to the suggestion that zeugmatin might be responsible for the fusion of the Z-bodies to form the solid Z-bands of the mature myofibrils (Rhee et al. 1994). As part of a study to test aspects of this model of myofibrillogenesis, we isolated a 1.8 kb cDNA from a chicken cardiac expression library using an anti-zeugmatin antibody (Turnacioglu et al., 1996). We found this chicken cDNA to be 60% identical at the amino acid level to a segment of the Z-band region of human cardiac titin (connectin) sequenced by Labeit and Kolmerer (1995). This homology along with Western blot analysis with purified titin, suggested that zeugmatin is in fact part of the N-terminal region of chicken titin. When expressed in non-muscle cells, Z1.1 product colocalized with the alpha-actinin in stress fiber dense bodies and focal adhesions. Cultures of non-muscle cells, skeletal myotubes and cardiomyocytes were also transfected with a fusion construct (Z1.1GFP) consisting of the Z1.1 kb cDNA linked to the cDNA for green fluorescent protein (GFP). The Z1.1 kb cDNA encodes only 362 of the approximately 2,000 amino acids which comprise the Z-band region of titin; nevertheless, the Z1.1GFP fusion protein targets in vivo to the alpha-actinin rich Z-bands of contracting myofibrils. A dominant negative phenotype was observed in living cells expressing high levels of this Z1.1GFP fusion protein with inhibition of myofibrillogenesis as well as the disassembly of preexisting myofibrils in these cells. These data indicate that the Z-band region of titin (connectin) plays an important role in organizing and maintaining the structure of the myofibril.


AID and TET2 co-operation modulates FANCA expression by active demethylation in diffuse large B cell lymphoma.
Diffuse large B cell lymphoma (DLBCL) is traced to a mature B malignance carrying abnormal activation-induced cytidine deaminase (AID) expression. AID activity initially focuses on deamination of cytidine to uracil to generate somatic hypermutation and class-switch recombination of the immunoglobulin (Ig), but recently it has been implicated in DNA demethylation of genes required for B cell development and proliferation in the germinal centre (GC). However, whether AID activity on mutation or demethylation of genes involves oncogenesis of DLBCL has not been well characterized. Our data demonstrate that the proto-oncogene Fanconi anaemia complementation group A (FANCA) is highly expressed in DLBCL patients and cell lines, respectively. AID recruits demethylation enzyme ten eleven translocation family member (TET2) to bind the FANCA promoter. As a result, FANCA is demethylated and its expression increases in DLBCL. On the basis of our findings, we have developed a new therapeutic strategy to significantly inhibit DLBCL cell growth by combination of the proteasome inhibitor bortezomib with AID and TET2 depletion. These findings support a novel mechanism that AID has a crucial role in active demethylation for oncogene activation in DLBCL.


Child Labor in Family Tobacco Farms in Southern Brazil: Occupational Exposure and Related Health Problems.
Tobacco farming is considered Hazardous Child Labor in Brazil. This study examined the work of children and adolescents in tobacco farming, characterizing the level of urinary cotinine and the occurrence of Green Tobacco Sickness (GTS), pesticide poisoning, respiratory symptoms, and musculoskeletal disorders. A cross-sectional descriptive study was conducted with a random sample of tobacco growers under 18 years old in Southern Brazil. Ninety-nine young people were interviewed at 79 family farms. The majority began working in agriculture before they were 14 and worked harvesting and tying hands of tobacco; 60% were 16 or 17 years old, and 51.5% were male. During their lifetime, 24.5% reported GTS, and 3% reported pesticide poisoning. In the previous year, 29.3% reported low back pain, 6.1% wheezing, and 16.2% coughing without having a cold. Half of the 12 young people evaluated had over 100 ng/mL of urinary cotinine. The study indicates that child laborers do various activities and present a high prevalence of health problems. Health workers should be trained to identify child laborers and their impacts on health. Full-time farm schools could provide knowledge about sustainable agricultural production, reducing the rates of age-grade mismatch, without taking young people away from rural areas.


Interaction of cholesterol-crystallization-promoting proteins with vesicles.
In this study, the interaction of mucin and concanavalin A-binding proteins isolated from human bile with cholesterol/phospholipid vesicles was investigated. Using resonance energy transfer assays originally developed by Struck, Hoekstra and Pagano [(1981) Biochemistry 20, 4093-4099], no significant protein-induced fusion or aggregation of vesicles was demonstrated. Instead of fusion, these proteins induced destabilization of cholesterol/phospholipid vesicles, as monitored by release of entrapped carboxyfluorescein. A good correlation (rho = 0.81) was obtained between the extent of leakage and the nucleation-promoting activity of the concanavalin A-binding proteins. We conclude that aggregation or fusion of cholesterol/phospholipid vesicles is not an obligatory step in cholesterol crystallization. Biliary protein-induced crystallization seems to be preceded by vesicle disruption.


Fast implementations of reconstruction-based scatter compensation in fully 3D SPECT image reconstruction.
Accurate scatter compensation in SPECT can be performed by modelling the scatter response function during the reconstruction process. This method is called reconstruction-based scatter compensation (RBSC). It has been shown that RBSC has a number of advantages over other methods of compensating for scatter, but using RBSC for fully 3D compensation has resulted in prohibitively long reconstruction times. In this work we propose two new methods that can be used in conjunction with existing methods to achieve marked reductions in RBSC reconstruction times. The first method, coarse-grid scatter modelling, significantly accelerates the scatter model by exploiting the fact that scatter is dominated by low-frequency information. The second method, intermittent RBSC, further accelerates the reconstruction process by limiting the number of iterations during which scatter is modelled. The fast implementations were evaluated using a Monte Carlo simulated experiment of the 3D MCAT phantom with 99mTc tracer, and also using experimentally acquired data with 201Tl tracer. Results indicated that these fast methods can reconstruct, with fully 3D compensation, images very similar to those obtained using standard RBSC methods, and in reconstruction times that are an order of magnitude shorter. Using these methods, fully 3D iterative reconstruction with RBSC can be performed well within the realm of clinically realistic times (under 10 minutes for 64 x 64 x 24 image reconstruction).


Isotope effect in superconducting fullerenes.
The effect of isotopic substitution on the superconducting transition temperature, T(c), in alkali-doped C(60) has been examined. Paradoxically, it is found that a substantial decrease in T(c) with the increasing isotopic mass is possible even when the attractive interaction is not mediated by phonons but is instead of purely electronic origin. In particular, it is shown that the experimentally measured isotopic shifts are consistent with a recently proposed electronic mechanism. Further predictions are presented that can be tested by experiment.


Biosecurity and bird movement practices in upland game bird facilities in the United States.
Since 1996, the emergence of Asian-origin highly pathogenic avian influenza subtype H5N1 has spurred great concern for the global poultry industry. In the United States, there is concern over the potential of a foreign avian disease incursion into the country. Noncommercial poultry operations, such as upland game bird facilities in the United States, may serve as a potential source of avian disease introduction to other bird populations including the commercial poultry industry, backyard flocks, or wildlife. In order to evaluate how to prevent disease transmission from these facilities to other populations, we examined biosecurity practices and bird movement within the upland game bird industry in the United States. Persons that held a current permit to keep, breed, or release upland game birds were surveyed for information on biosecurity practices, flock and release environments, and bird movement parameters. Biosecurity practices vary greatly among permit holders. Many facilities allow for interaction between wild birds and pen-reared birds, and there is regular long-distance movement of live adult birds among facilities. Results suggest that upland game bird facilities should be targeted for biosecurity education and disease surveillance efforts.


Degradation characteristics of two Bacillus strains on the Microcystis aeruginosa.
The degradation kinetics of strains P05 and P07 and the degradation effects of mixed strain on Microcystis aeruginosa were studied. The results showed that: (1) The degradation processes of strains P05 and P07 on Microcystis aeruginosa accorded with the first-order reaction model when the range of Chl-a concentration was from 0 to 1500 microg/L. (2) The initial bacterium densities had a strong influence on the degradation velocity. The greater the initial bacterium density was, the faster the degradation was. The degradation velocity constants of P05 were 0.1913, 0.2175 and 0.3092 respectively, when bacterium densities were 4.8 x 10(5), 4.8 x 10(6), 2.4 x 10(7) cells/ml. For strain P07, they were 0.1509, 0.1647 and 0.2708. The degradation velocity constant of strain P05 was higher than that of P07 when the bacterium density was under 4.8 x 10(5) cells/ml, but the constant increasing of P07 was quicker than that of P05. (3) The degradation effects of P05 and P07 strains did not antagonize. When the concentration of Chl-a was high, the degradation effects of mixed strain excelled that of any single strains. But with the decrease of the Chl-a concentration, this advantage was not clear. When the concentration was less than 180 microg/L, the degradation effects of mixed were consistent with that of strain P07.


Safety and tolerability of ceftobiprole.
Ceftobiprole is a fifth generation cephalosporin with a series of characteristics differentiating it from other beta-lactams, including its antibacterial activity, mainly against methicillin-resistant Staphylococcus aureus, resistant Streptococcus pneumoniae and also Gram-negative microorganisms such as Pseudomonas aeruginosa. This antibiotic has been subjected to various clinical trials and the results of these have led to its approval in Spain for the treatment of nosocomial pneumonia, excluding that associated with mechanical ventilation, and community-acquired pneumonia. The results of various ceftobiprole clinical studies provide consistent information on efficacy and tolerability. Ceftobiprole as monotherapy has been shown to be non-inferior to comparator antibiotics in different settings. Information is available on its compatibility with other drugs in Y-site administration, important from the point of view of the intravenous treatment of patients who present venous access limitation. On the other hand, and in contrast to other cephalosporins, ceftobiprole presents a low risk of infection due to Clostridium difficile and, in comparison with ceftaroline, neutropenia has not been reported to present any significant issues.


Effects of a new antianginal agent, nicorandil, on the cardiovascular system of the miniature pig: with special reference to coronary vasospasm.
The cardiovascular and vasospasmolytic effects of a newly developed nicotinamide derivative, N-(2-hydroxyethyl)nicotinamide nitrate(ester) (nicorandil), were examined in anesthetized, open-chest "miniature" pigs, and compared with those of nitroglycerin. Nicorandil, administered i.v. in doses of 10 micrograms/kg to 1 mg/kg, decreased systemic blood pressure, left ventricular (LV) systolic pressure, pressure rate product and coronary and femoral vascular resistance. It pronouncedly increased coronary blood flow and slightly increased heart rate. An increase, often preceded by a decrease, occurred in the LVdP/dt max. In doses of 10 to 100 micrograms/kg, nicorandil induced a slight increase followed by a decrease in femoral blood flow, but larger doses always decreased it. Similar results were obtained with nitroglycerin for all the cardiovascular parameters tested except LVdP/dt max and coronary blood flow. Unlike nicorandil, nitroglycerin induced a decrease often preceded by an increase in the LVdP/dt max. In doses of 1 to 30 micrograms/kg, nitroglycerin caused a decrease preceded by a slight and transient increase in the coronary blood flow. Nicorandil, administered i.v. in doses of 30 micrograms/kg to 1 mg/kg, inhibited coronary vasoconstriction caused by methacholine (3 or 10 micrograms) and norepinephrine (2-4 micrograms in the presence of propranolol) injected directly into the coronary artery. The magnitude and duration of this antagonism depended on the dose of nicorandil used. Although nitroglycerin in a dose of 10 or 30 micrograms/kg also effectively inhibited coronary vasoconstrictor responses to the two vasoactive agents, dipyridamole (60 micrograms/kg) failed to do so. Thus, nicorandil has prominent vasospasmolytic effects and is an efficacious coronary vasodilator which reduces afterload.


Ultrastructure alteration of decidual natural killer cells in women with unexplained recurrent miscarriage: a possible association with impaired decidual vascular remodelling.
This study aimed to evaluate the extent of remodelling of intra-decidual segments of the spiral arteries in human deciduas between the 6th and 10th gestational weeks in women with unexplained recurrent miscarriages (RM) in comparison to gestational-matched controls. A possible association with the number, immunoexpressive behaviour and ultrastructural changes of decidual natural killer cells (dNKCs) was investigated. Decidual biopsies were obtained from RM cases (n = 40) and women with no history of spontaneous miscarriage and at least one live birth at term (n = 30). Staining was performed using PAS, anti-CD34 and anti-CD56 antibodies, using an avidin-biotin-peroxides technique. Analysis by means of light and transmission electron microscopy was employed. To determine the extent of remodelling of decidual vessels, a quantitative score was analysed using histological criteria of vascular transformation and then related to the number of CD56(+) dNKCs. In RM, dNKCs were distributed among decidual cells and around the vessels. They possessed numerous polyploidic protrusions on cell membranes crossing from one cell to another. The cells became more irregular and exhibited heterogeneous electron-dense granules in their cytoplasm compared to controls. The non-remodelling score and number of dNKCs were significantly increased in RM group (p < 0.001). The number of dNKCs was significantly correlated with the scores in both control (r = 0.491; p = 0.006) and RM (r = 0.852; p < 0.001) groups. It appears that dNKCs play a key role in impaired decidual artery remodelling that may be involved with early RM. This may be due to increased numbers of cells or impaired cellular interactions resulting from alterations to the ultrastructure.


SPATIAL VARIATION OF WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) POPULATION IMPACTS AND RECOVERY FROM EPIZOOTIC HEMORRHAGIC DISEASE.
Epizootic hemorrhagic disease (EHD) is the most significant source of viral disease-related mortality in white-tailed deer (Odocoileus virginianus) in the US. Deer mortality from EHD has increased in the state of Michigan, US, since 2006, with the largest outbreak occurring in 2012. The 2012 outbreak provided an opportunity to evaluate how this disease affected EHD-related mortality in deer populations at a spatial scale typical of that expected for the greatest disease risk. Our objectives were to quantify the population impacts and spatial extent of EHD associated with areas of disease risk for deer populations and to determine how populations recovered over time following localized EHD impacts. We estimated the annual local abundance of deer for 5 yr immediately following a recent EHD outbreak. Because proximity to wetlands may affect EHD occurrence, we surveyed deer at varying distances (about 1 km and 5 km) from a riparian corridor to determine spatial variation in population impacts. Further, we assessed differences in deer abundance for sites affected and unaffected by EHD. Abundance estimates were lower along transects near the riparian corridor only in the affected area, reflecting EHD mortality associated with wetlands. The only change in abundance over time was a significant increase in the riparian strata in the EHD-affected site.


Purification and characterization of the class-II D-fructose 1,6-bisphosphate aldolase from Escherichia coli (Crookes' strain).
A new form of the class-II D-fructose 1,6-bisphosphate aldolase (EC 184.108.40.206) of Escherichia coli (Crookes' strain) was isolated from an extract of glycerol-grown bacteria. It has a higher molecular weight (approx. 80000)than previous preparations of the enzyme and closely resembles the typical class-II aldolase from yeast in size and amino acid composition. On the other hand, its kinetic behaviour is not typical of a class-II aldolase. The enzyme has no requirement for thiol compounds either for stability or activity, added K+ ions have no effect, and the optimum pH for the cleavage activity is unusually high. The class-II enzymes from the prokaryote E. coli and the eukaryote yeast show no immunological identity. However, the similarity of their structures suggests that they have evolved from a common ancestor.


[Association between calcium content of drinking water and fractures in children].
The goal of this research was to determine the relationship between high levels of calcium in Majorca's water supplies and the prevalence of fractures in schoolchildren. The study was performed during two school terms and was done within the framework of the School Health System. Three towns of Majorca were selected because of the different content of calcium in their water supply. The fluorine content in the tap water was always under 0.5 mg/l. All of the towns have the same social and economic backgrounds. We surveyed 1,308 schoolchildren, between 11 and 14 years of age, about previous fractures. There is a statistically significant relationship between fractures and low calcium levels in the water supply. This association is stronger for children 11 years of age than for children 14 years old.


Grading criteria for chronic ocular graft-versus-host disease: comparing the NIH eye score, Japanese dry eye score, and DEWS 2007 score.
Ocular graft-versus-host disease (GVHD) is a common complication after hematopoietic stem cell transplantation (HSCT). Here we compared the diagnostic rates of ocular GVHD, including its severity, prognosis and the agreement, obtained using three grading scales: the National Institutes of Health (NIH) eye score, Japanese dry eye score, and dry eye workshop score, by retrospectively reviewing the records of 82 patients who underwent HSCT. Tear dynamics and ocular surface assessments made 6-9 months after HSCT were used to determine ocular GVHD severity with the three scales. By the three scales, ocular GVHD was diagnosed in 56 patients (68.3%), 51 patients (62.2%), and 52 patients (63.4%), respectively. The Kappa coefficient was calculated to determine the agreement between scales for diagnosing ocular GVHD. The severity progression within two years after onset was also assessed by tear dynamics and ocular surface examination. The patients were categorized as no change, improved, or progressive. The three grading scales showed good agreement (Kappa = 0.87 to 0.97) in diagnosing patients with ocular GVHD, and the scores by all three were significantly associated with the patients' prognosis (p < 0.01). We recommend that multi-center research is needed for further validation and investigation.


How reproducible are transcranial magnetic stimulation-induced MEPs in subacute stroke?
Motor evoked potentials (MEPs) and total motor conduction time (TMCT) induced by transcranial magnetic stimulation (TMS) are used to make assumptions about the prognosis of motor outcome after stroke. Understanding the different sources of variability is fundamental to the concept of reliability. Reliability testing of TMS-MEPs and TMCTs within and between two independent examiners in healthy and stroke subjects is still an unexplored field in the clinical neurophysiology. Assessing the reproducibility of TMS measurements requires studies to investigate the test-retest reliability of TMS-induced MEPs and TMCT. The authors set out to test the reliability of these TMS measurements. Eighteen patients with stroke and 8 healthy volunteers were tested twice within a 1-week period by 2 examiners using TMS to determine MEPs and TMCT for the abductor pollicis brevis muscle of their affected and unaffected hands. The authors found moderate to perfect reliability of TMS-induced MEPs in healthy volunteers, noninfarcted hemispheres (perfect agreement), and infarcted hemispheres (Kappa's = 0.45-0.87). Reliability of TMCT was good to excellent in the volunteers (intraclass correlation coefficients = 0.77-0.97), excellent in the noninfarcted hemispheres (intraclass correlation coefficients = 0.97-1.00), and poor to excellent in the infarcted hemispheres (intraclass correlation coefficients = 0.44-0.90). The reliability of TMS-induced MEPs and TMCT measurements in healthy volunteers and the noninfarcted hemisphere of patients with stroke with an upper paretic limb was good to excellent. In contrast, TMS measurements in the infarcted hemisphere were less consistent. Based on the lower reproducibility of TMCT measurements in the infarcted hemisphere, we recommend to repeat the TMCT measurements to improve the reliability of tests.


Molecular detection of uncultured cyanobacteria and aminotransferase domains for cyanotoxin production in sediments of different Kenyan lakes.
PCR-based denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments was used to identify the cyanobacterial phylotypes in sediments and plankton of saline-alkaline and freshwater lakes of Kenya. The detection of the aminotransferase domain located on modules mcyE and ndaF using specific molecular markers confirmed the presence of potential toxin-producing cyanobacteria. The eight nucleotide sequences obtained from DGGE bands were placed in three divergent cyanobacterial clusters. Five nucleotide sequences were close to members of the genera Anabaenopsis and Umezakia (Nostocales), two sequences fell in the cluster with Arthrospira sp. (Oscillatoriales) and one sequence was related to Chroococcidiopsis sp. (Pleurocapsales). The presence of the latter taxon was demonstrated de novo in the investigated lakes. All nine attained nucleotide sequences of the aminotransferase region belonged to the mcyE module. Five sequences of the aminotransferase domain were included in the cluster having the nucleotide sequence of Anabaena sp. but showed a separate lineage. Other four aminotransferases were placed in the cluster represented by nucleotide sequence of Microcystis aeruginosa. To our knowledge, this is the first report on molecular detection of cyanobacterial phylotypes in sediments of African lakes and aminotransferase domains for cyanotoxin production from sediment samples in general.


[Lateral cervical puncture as a new method for collecting cerebrospinal fluid. Experience in Recife and its metropolitan area].
200 cervical lateral punctions (CLP) were performed in comatose patients without psychomotor trouble proving this new system to be an alternative for suboccipital punction. This procedure was successful in 90% of the cases. Low importance venous accidents happened but they did not bring any injury or serious consequences to the patients or to cerebrospinal fluid examination results. Vertebral artery injury was not observed. Punctions in corpses showed us that this artery is one cm far from the needle trajectory during the realization of the CLP.


Communication Architecture in Mixed-Reality Simulations of Unmanned Systems.
Verification of the correct functionality of multi-vehicle systems in high-fidelity scenarios is required before any deployment of such a complex system, e.g., in missions of remote sensing or in mobile sensor networks. Mixed-reality simulations where both virtual and physical entities can coexist and interact have been shown to be beneficial for development, testing, and verification of such systems. This paper deals with the problems of designing a certain communication subsystem for such highly desirable realistic simulations. Requirements of this communication subsystem, including proper addressing, transparent routing, visibility modeling, or message management, are specified prior to designing an appropriate solution. Then, a suitable architecture of this communication subsystem is proposed together with solutions to the challenges that arise when simultaneous virtual and physical message transmissions occur. The proposed architecture can be utilized as a high-fidelity network simulator for vehicular systems with implicit mobility models that are given by real trajectories of the vehicles. The architecture has been utilized within multiple projects dealing with the development and practical deployment of multi-UAV systems, which support the architecture's viability and advantages. The provided experimental results show the achieved similarity of the communication characteristics of the fully deployed hardware setup to the setup utilizing the proposed mixed-reality architecture.


The effect of surgery time on prognosis in newborns with meningomyelocele.
To investigate the effect of surgery time on prognosis of newborns with meningomyelocele. The records of neonates with meningomyelocele were retrospectively analyzed. Demographic and clinical characteristics as well as information, timing of surgery, and durations of hospital stay and antibiotic therapy were recorded. The records of 30 babies were included in the final analysis. Overall, the mean gestational age was 37.7±2.7 weeks, with a mean birth weight of 2967±755 g and head circumference of 35.8±3.8 cm. In terms of localization, 46.6% of the meningomyeloceles were lumbosacral, 40% were lumbar, 10% were thoracolumbar and 3.3% were thoracal. The mean size of the meningomyelocele sacs was 4.33±1.2 cm. Newborns underwent surgery on average of 8.2±5.9 days after birth, with an overall mean duration of hospital stay of 30±25.1 days. Patients were divided into two groups based on timing of surgery (group 1, ≤5 days; group 2, >5 days), and comparisons between groups revealed that earlier surgery was associated with significantly shorter durations of hospital stay (p<0.001) and antibiotic therapy (p<0.05). Early surgical intervention (≤5 days) was associated with a shorter duration of hospital stay and antibiotic therapy as well as a lower complication rate. We recommend that corrective surgery be undertaken as soon as reasonably possible.


Nonlinear optical studies of the agricultural antibiotic morantel interacting with silica/water interfaces.
It is now known that the untreated discharge of pharmaceuticals into the environment can impact human health and development and lead to increased drug resistance in biota. Here, we present the first direct interface-specific studies that address the mobility of the widely used agricultural antibiotic morantel, which is commonly present in farm runoff. Surface-bound morantel was spectroscopically identified using second harmonic generation (SHG) via a two-photon resonance of its n-pi* transition and in the C-H stretching region by vibrational sum frequency generation (VSFG). Resonantly enhanced SHG adsorption isotherm measurements carried out at the silica/water interface between 6 x 10(-7) and 5 x 10(-5) M morantel concentration result in a free energy of adsorption of 42(2) kJ/mol at pH 7. Finally, real-time tracking of morantel interaction with the silica/water interface shows that the binding events are fully reversible, consistent with its high mobility in silica-rich soil environments. This work thus indicates that pharmaceuticals discharged into the environment can enter the groundwater supply of municipal water systems, at which point their removal is challenging. In addition, the high mobility of morantel in silica-rich soil environments could lead to developing increased interaction of this antibiotic with target organisms, which could respond by increased drug resistance.


Regulation of telomeric function by DNA methylation differs between humans and mice.
The most distal 2 kb region in the majority of human subtelomeres contains CpG-rich promoters for TERRA, a long non-coding RNA. When the function of the de novo DNA methyltransferase DNMT3B is disrupted, as in ICF1 syndrome, subtelomeres are abnormally hypomethylated, subtelomeric heterochromatin acquires open chromatin characteristics, TERRA is highly expressed, and telomeres shorten rapidly. In this study, we explored whether the regulation of subtelomeric epigenetic characteristics by DNMT3B is conserved between humans and mice. Studying the DNA sequence of the distal 30 kb of the majority of murine q-arm subtelomeres indicated that these regions are relatively CpG-poor and do not contain TERRA promoters similar to those present in humans. Despite the lack of human-like TERRA promoters, we clearly detected TERRA expression originating from at least seven q-arm subtelomeres, and at higher levels in mouse pluripotent stem cells in comparison with mouse embryonic fibroblasts (MEFs). However, these differences in TERRA expression could not be explained by differential methylation of CpG islands present in the TERRA-expressing murine subtelomeres. To determine whether Dnmt3b regulates the expression of TERRA in mice, we characterized subtelomeric methylation and associated telomeric functions in cells derived from ICF1 model mice. Littermate-derived WT and ICF1 MEFs demonstrated no significant differences in subtelomeric DNA methylation, chromatin modifications, TERRA expression levels, telomere sister chromatid exchange or telomere length. We conclude that the epigenetic characteristics of murine subtelomeres differ substantially from their human counterparts and that TERRA transcription in mice is regulated by factors others than Dnmt3b.


Neuropsychiatric phenotypes in functional movement disorder.
Functional movement disorder (FMD), the motor-dominant subtype of functional neurological disorder, is a complex neuropsychiatric condition. Patients with FMD also manifest non-motor symptoms. Given that patients with FMD are diagnosed based on motor phenotype, the contribution of non-motor features to the neuropsychiatric syndrome is not well characterized. The objective of this hypothesis-generating study was to explore potential novel, neuropsychiatric FMD phenotypes by combining movement disorder presentations with non-motor comorbidities including somatic symptoms, psychiatric diagnoses, and psychological traits. This retrospective chart review evaluated 158 consecutive patients with a diagnosis of FMD who underwent deep phenotyping across neurological and psychiatric domains. Demographic, clinical, and self-report features were analyzed. A data-driven approach using cluster analysis was performed to detect patterns when combining the movement disorder presentation with somatic symptoms, psychiatric diagnoses, and psychological factors. These new neuropsychiatric FMD phenotypes were then tested using logistic regression models. Distinct neuropsychiatric FMD phenotypes emerged when stratifying by episodic vs. constant motor symptoms. Episodic FMD was associated with hyperkinetic movements, hyperarousal, anxiety, and history of trauma. In contrast, constant FMD was associated with weakness, gait disorders, fixed dystonia, activity avoidance, and low self-agency. Pain, fatigue, somatic preoccupation, and health anxiety were common across all phenotypes. This study found patterns spanning the neurological-psychiatric interface that indicate that FMD is part of a broader neuropsychiatric syndrome. Adopting a transdisciplinary view of illness reveals readily identifiable clinical factors that are relevant for the development and maintenance of FMD.


TRIM-mediated precision autophagy targets cytoplasmic regulators of innate immunity.
The present paradigms of selective autophagy in mammalian cells cannot fully explain the specificity and selectivity of autophagic degradation. In this paper, we report that a subset of tripartite motif (TRIM) proteins act as specialized receptors for highly specific autophagy (precision autophagy) of key components of the inflammasome and type I interferon response systems. TRIM20 targets the inflammasome components, including NLRP3, NLRP1, and pro-caspase 1, for autophagic degradation, whereas TRIM21 targets IRF3. TRIM20 and TRIM21 directly bind their respective cargo and recruit autophagic machinery to execute degradation. The autophagic function of TRIM20 is affected by mutations associated with familial Mediterranean fever. These findings broaden the concept of TRIMs acting as autophagic receptor regulators executing precision autophagy of specific cytoplasmic targets. In the case of TRIM20 and TRIM21, precision autophagy controls the hub signaling machineries and key factors, inflammasome and type I interferon, directing cardinal innate immunity response systems in humans.


[Long-term results of surgical treatment of patent ductus arteriosus in adults].
284 patients with patent ductus arteriosus were operated. Among them, 81 aged from 20-41. Special managements were made because of their serious pathologic changes. There was no operative mortality. 72 patients (88.9%) were followed up for 1-17 years. The results of follow-up indicated that the postoperative cardiopulmonary function improved greatly. 63 patients returned normal work. The long-term results were satisfactory.


Benefits of a Nationwide Palliative Care Education Program on Lung Cancer Physicians.
Objective The early integration of palliative care into standard cancer treatment has become a global standard. The Palliative care Emphasis program on symptom management and Assessment for Continuous medical Education (PEACE) has been conducted in Japan, and previous studies have reported that the PEACE workshop was able to improve various palliative care skills of participants. However, whether or not the effects of the program are long-lasting and if the program consequently changed physicians' practice with regard to lung cancer patients have been unclear. Methods Web-based surveys, including the palliative care knowledge test (PEACE-Q), the Palliative Care self-reported Practice Scale (PCPS), and the Palliative Care Difficulties Scale (PCDS), were conducted among lung cancer physicians in Japan. The differences in the survey results between participants and non-participants of the PEACE workshop were examined. Results Among 923 respondents (455 respiratory physicians, 345 pulmonary surgeons, and 123 others), 519 had participated in the PEACE workshop. The total PEACE-Q score was significantly higher in the PEACE workshop participants than in non-participants (28.0 versus 24.5, p<0.0001). The score was significantly higher in respiratory physicians than in pulmonary surgeons (27.4 versus 25.5). The total PCPS and PCDS scores were also significantly better in workshop participants than in non-participants (71.8 versus 67.1 and 34.3 versus 36.9, respectively), although some domains of PCDS were similar between the groups. Conclusion The PEACE program improved the knowledge and practices with regard to palliative care and resolved difficulties associated therewith among lung cancer physicians. In regions where palliative care specialists are insufficient, such educational programs may be effective.


How should we interpret noncompliance with screening mammography?
Primary care practitioners have an important role to play in recommending breast cancer screening to patients in the target age group. In this issue of CMAJ (see pages 1335 to 1343) Dr. Marie-Dominique Beaulieu and associates report the results of a program designed to maximize utilization of screening mammography. Only two thirds of eligible women for whom screening mammography was prescribed obtained a mammogram within the 2-month study period. However, when taken in context, this compliance rate is fairly encouraging. There are many possible reasons for noncompliance such as a need for more information or for repeated suggestions. Family physicians should not become disheartened in their efforts to increase the use of screening procedures and may find that collaboration with others in giving consistent messages will help to maximize screening rates within their patient population.


Gonadal toxicity of short term chronic endosulfan exposure to male rats.
Endosulfan was studied for its effect on rat testicular toxicity in relation to the enzymes of androgen biosynthesis, viz. 3 beta-hydroxysteroid dehydrogenase (EC 220.127.116.11, 3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (EC 22.214.171.124, 17 beta-HSD); cytosolic conjugation enzyme, glutathione-S-transferase (EC 188.8.131.52); and testicular as well as serum testosterone levels at the dose levels of 2.5, 5.0, 7.5 and 10 mg/kg body weight fed orally for 7 and 15 days. Organ and body weights of the treated animals did not change significantly, however, the testicular protein contents were found to be increased appreciably after 7 days treatments. The activity profile of cytosolic conjugation enzyme showed much remained low during 7 days treatment, however, the two steroidogenic enzymes showed much individual variations in response to endosulfan treatments. An overall varied response with respect to testosterone biosynthesis and its secretion to serum was observed suggesting nevertheless, a profound hormonal imbalance caused by this insecticide to male gonads on short term chronic exposures.


The Injury Profile of an Australian Specialist Policing Unit.
This study investigated the injuries sustained by an Australian specialist police division. Injury records spanning four-years were analyzed. The role being performed when the injury occurred, injury cause, body part injured, and injury-related costs were quantified. The percentage of personnel injured multiple times was documented. One hundred and thirty eight personnel reported injuries, 58 of these on multiple occasions. This resulted in 229 injuries and 76 claims being raised. Half of the injuries occurred during operational policing tasks, however training activities accounted for >30% of injuries. The most common injury was strain/sprain, and upper body injuries were 2.5-times more common than lower-body or torso injuries. 1107 shifts were lost, and injuries cost the organization $487,159 (Australian Dollars) over the four-year period. The injury costs (both financial and in manpower) may prompt policy makers to review the current training and post-injury rehabilitation protocols.


Acute subdural hematomas. Review of 144 cases.
The authors report 144 cases in which acute subdural hematomas resulting from closed head injury were surgically treated. The mortality rate was 48% for those treated within 24 hours of injury and 45% for those treated within 72 hours. Patients under 10 years of age had a 33% mortality, while 69% of those over 60 years died. In the first 6 years of the series, 75% of the patients were treated by multiple burr holes and a subtemporal craniectomy, with a mortality of 41% in the last 6 years, 92% of the patients had large craniotomies with a 45% mortality. Of the 32 survivors among the last 60 patients treated, nine require full nursing home care, eight have returned to their own homes, 12 are able to care for themselves but not work, and three have returned to work.


TissueEnrich: Tissue-specific gene enrichment analysis.
RNA-Seq data analysis results in lists of genes that may have a similar function, based on differential gene expression analysis or co-expression network analysis. While tools have been developed to identify biological processes that are enriched in the genes sets, there remains a need for tools that identify enrichment of tissue-specific genes. Therefore, we developed TissueEnrich, a tool that calculates tissue-specific gene enrichment in an input gene set. We demonstrated that TissueEnrich can assign tissue identities to single cell clusters and differentiated embryonic stem cells. The TissueEnrich web application is freely available at http://tissueenrich.gdcb.iastate.edu/. The R package is available through Bioconductor at https://bioconductor.org/packages/TissueEnrich. Both the web application and R package are for non-profit academic use under the MIT license. Supplementary data are available at Bioinformatics online.


Ginsenoside Rg3 Decreases Fibrotic and Invasive Nature of Endometriosis by Modulating miRNA-27b: In Vitro and In Vivo Studies.
This research aimed to evaluate the potential therapeutic effects of Rg3 on endometriosis and identify target miRNAs. We designed an in vitro study using human endometrial stromal cells (HESCs) obtained from patients with endometriosis and an in vivo study using mouse models. HESCs were treated with Rg3-enhanced red ginseng extract (Rg3E); real-time PCR and microarray profiling, transfection, and western blot were performed. Mouse endometriosis models were developed and supplemented with Rg3E for 8 weeks. Gross lesion size and fibrotic character were analyzed in the mouse models. RNA levels of Ki-67, col-1, CTGF, fibronectin, TGF-β1, MMP2 and MMP9 significantly decreased in HESCs after Rg3E treatment. Microarray analysis revealed downregulation of miR-27b-3p, which is related to fibrosis modulation. Expression of miR-27b-3p was significantly higher in HESCs from patients with endometriosis than that of controls, and Rg3E treatment significantly decreased its expression; the contraction and migration assay revealed significant reductions in both fibrosis and migration potential in Rg3E-treated HESCs from endometriosis patients. A decrease in size and fibrotic character of endometrial lesions from the Rg3E groups was observed in vivo. In conclusion, Rg3 effectively altered fibrotic properties of HESCs from patients with endometriosis, which is likely associated with miR-27b-3p modulation.


[Is H15 (resin extract of Boswellia serrata, "incense") a useful supplement to established drug therapy of chronic polyarthritis? Results of a double-blind pilot study].
Leukotrienes and prostaglandines are important mediators of inflammation. While prostaglandine synthesis can be influenced by NSAIDs therapeutical approaches to the 5-lipoxygenase pathway are rare. Resinous extracts of Boswellia serrata (H15, indish incense), known from traditional ayurvedic medicine, decrease leukotriene synthesis in vitro. Case reports suggest a clinical role for that drug. Outpatients with active RA have been enrolled into a multicenter controlled trial. Patients received 9 tablets of active drug (3600 mg) or placebo daily in addition to their previous therapy. Doses of NSAIDs could be adjusted on demand. Efficacy parameters, Ritchies Index for swelling and pain, ESR, CRP, pain on VAS and NSAID dose were documented at baseline and 6 and 12 weeks after initiation. Mean values and medians were calculated to compare the groups for significant or clinically relevant change from baseline or difference between both groups at any time point of observation. A total of 78 patients were recruited in 4 centers, the data have been published in abstract form. Only 37 patients (verum 18, placebo 19), enrolled in Ratingen were available for detailed efficacy and safety analysis. All evaluations in these patients were performed by one investigator (G.H.). There was no subjective, clinical or laboratory parameter showing a significant or clinically relevant change from baseline or difference between both groups at any time point of observation. The mean NSAID dose reduction reached levels of 5.8% (H15) and 3.1% (placebo). One patient in each group showed a good response in all parameters but 4 patients in each group worsened. The others showed no alteration of their disease. Treatment with H15 showed no measurable efficacy. Controlled studies including a greater patient population are necessary to confirm or reject our results.


Association of Interleukin-28B Polymorphisms (rs12979860 C/T, rs12980275 A/G, rs8099917 T/G) and Risk of Hepatocellular Carcinoma in an Iranian Population.
BACKGROUND: Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. The association of interleukin 28B (IL-28B) polymorphisms and HCC has been investigated in several populations. However, the findings are controversial. This study aimed to address the association between IL-28B polymorphisms (rs 8099917 T/G, rs12979860 C/T, rs12980275 A/G) and the risk of HCC in an Iranian population. METHODS: We have evaluated the association between IL-28B polymorphisms (rs 8099917 T/G, rs12979860 C/T, rs12980275 A/G) and HCC in 180 Iranian individuals (60 patients with HCC and 120 healthy matched controls) using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) method. Single nucleotide polymorphism (SNP) association analysis and also haplotypes were estimated using the SNPstats online software. RESULTS: There was no significant association between these three polymorphisms of IL-28B and HCC (P>0.05). Moreover, haplotype analysis showed no significant association between the haplotypes and HCC. CONCLUSION: There was no association between IL-28B polymorphisms and HCC in an Iranian population.


Molecular biology of normal melanocytes and melanoma cells.
Malignant melanoma is one of the most aggressive malignancies in humans and is responsible for 60-80% of deaths from skin cancers. The 5-year survival of patients with metastatic malignant melanoma is about 14%. Its incidence has been increasing in the white population over the past two decades. The mechanisms leading to malignant transformation of melanocytes and melanocytic lesions are poorly understood. In developing malignant melanoma, there is a complex interaction of environmental and endogenous (genetic) factors, including: dysregulation of cell proliferation, programmed cell death (apoptosis) and cell-to-cell interactions. The understanding of genetic alterations in signalling pathways of primary and metastatic malignant melanoma and their interactions may lead to therapeutics modalities, including targeted therapies, particularly in advanced melanomas that have high mortality rates and are often resistant to chemotherapy and radiotherapy. Our knowledge regarding the molecular biology of malignant melanoma has been expanding. Even though several genes involved in melanocyte development may also be associated with melanoma cell development, it is still unclear how a normal melanocyte becomes a melanoma cell. This article reviews the molecular events and recent findings associated with malignant melanoma.


Quantitative characterization of disparity tuning in ventral pathway area V4.
We performed a quantitative characterization of binocular disparity-tuning functions in the ventral (object-processing) pathway of the macaque visual cortex. We measured responses of 452 area V4 neurons to stimuli with disparities ranging from -1.0 to +1.0 degrees. Asymmetric Gaussian functions fit the raw data best (median R = 0.90), capturing both the modal components (local peaks in the -1.0 to +1.0 degrees range) and the monotonic components (linear or sigmoidal dependency on disparity) of the tuning patterns. Values derived from the asymmetric Gaussian fits were used to characterize neurons on a modal x monotonic tuning domain. Points along the modal tuning axis correspond to classic tuned excitatory and inhibitory patterns; points along the monotonic axis correspond to classic near and far patterns. The distribution on this domain was continuous, with the majority of neurons exhibiting a mixed modal/monotonic tuning pattern. The distribution in the modal dimension was shifted toward excitatory patterns, consistent with previous results in other areas. The distribution in the monotonic dimension was shifted toward tuning for crossed disparities (corresponding to stimuli nearer than the fixation plane). This could reflect a perceptual emphasis on objects or object parts closer to the observer. We also found that disparity-tuning strength was positively correlated with orientation-tuning strength and color-tuning strength, and negatively correlated with receptive field eccentricity.


Stability and apoptotic activity of recombinant human cytochrome c.
An efficient system for producing human cytochrome c variants is important to help us understand the roles of this protein in biological processes relevant to human diseases including apoptosis and oxidative stress. Here, we describe an Escherichia coli expression system for producing recombinant human cytochrome c. We also characterize the structure, stability, and function of the protein and show its utility for studying apoptosis. Yields of greater than 8 mg of pure protein per liter culture were attained. Circular dichroism spectropolarimetry studies show that the secondary and tertiary structures of the human protein are nearly identical to those of the horse protein, but the human protein is more stable than other eukaryotic cytochromes c. Furthermore, recombinant human cytochrome c is capable of inducing caspase-3 activity in a cell-free caspase activation assay. We use data from this assay along with data from the literature to define the apaf-1 binding site on human cytochrome c.


Are the Healthy Vulnerable? Cytomegalovirus Seropositivity in Healthy Adults Is Associated With Accelerated Epigenetic Age and Immune Dysregulation.
Evaluating age as a risk factor for susceptibility to infectious diseases, particularly coronavirus disease 2019 (COVID-19), is critical. Cytomegalovirus (CMV) serologic prevalence increases with age and associates with inflammatory-mediated diseases in the elderly. However, little is known regarding the subclinical impact of CMV and risk it poses to healthy older adults. Prior to the COVID-19 pandemic we conducted a study to determine the association of CMV to biologic age and immune dysregulation. Community-dwelling, healthy adults older than 60 years were evaluated using DNA methylation assays to define epigenetic age (EpiAge) and T-cell immunophenotyping to assess immune dysregulation. All subjects were healthy and asymptomatic. Those CMV seropositive had more lymphocytes, CD8 T cells, CD28- T cells, decreased CD4:CD8 cell ratios, and had higher average EpiAge (65.34 years) than those CMV seronegative (59.53 years). Decreased percent CD4 (P = .003) and numbers of CD4 T cells (P = .0199) correlated with increased EpiAge. Our novel findings distinguish altered immunity in the elderly based on CMV status. Chronic CMV infection in healthy, older adults is associated with indicators of immune dysregulation, both of which correlate to differences in EpiAge.


Immunologic and endocrine functions of adipose tissue: implications for kidney disease.
Excess adiposity can induce adverse sequelae in multiple cell types and organ systems. The transition from the lean to the obese state is characterized by fundamental cellular changes at the level of the adipocyte. These changes affect the local microenvironment within the respective adipose tissue but can also affect nonadipose systems. Adipocytes within fat pads respond to chronic nutrient excess through hyperplasia or hypertrophy, which can differentially affect interorgan crosstalk between various adipose depots and other organs. This crosstalk is dependent on the unique ability of the adipocyte to coordinate metabolic adjustments throughout the body and to integrate responses to maintain metabolic homeostasis. These actions occur through the release of free fatty acids and metabolites during times of energy need - a process that is altered in the obese state. In addition, adipocytes release a wide array of signalling molecules, such as sphingolipids, as well as inflammatory and hormonal factors (adipokines) that are critical for interorgan crosstalk. The interactions of adipose tissue with the kidney - referred to as the adipo-renal axis - are important for normal kidney function as well as the response of the kidney to injury. Here, we discuss the mechanistic basis of this interorgan crosstalk, which clearly has great therapeutic potential given the increasing rates of chronic kidney disease secondary to obesity and type 2 diabetes mellitus.


Predictive value of antimicrobial susceptibility from previous urinary tract infection in the treatment of re-infection.
Recurrent urinary tract infections are often re-infections; therefore, antimicrobial susceptibility test results from a previous episode may guide the empiric therapy in subsequent episodes. This analysis provides predictive values of the antimicrobial susceptibility of previous Escherichia coli isolates for the treatment of re-infections in routine clinical practice. If resistance to ampicillin, trimethoprim, or ciprofloxacin is detected, re-prescription within 3 months is imprudent. Susceptibility to nitrofurantoin, ciprofloxacin, or trimethoprim in a previous sample supports their prescription for a re-infection within 3 months and up to a year. Resistance to nitrofurantoin is low and, once detected, decays relatively quickly. Nitrofurantoin should be considered as a first-line agent for initial and repeat treatment.


Attentional effects on local V1 microcircuits explain selective V1-V4 communication.
Selective attention implements preferential routing of attended stimuli, likely through increasing the influence of the respective synaptic inputs on higher-area neurons. As the inputs of competing stimuli converge onto postsynaptic neurons, presynaptic circuits might offer the best target for attentional top-down influences. If those influences enabled presynaptic circuits to selectively entrain postsynaptic neurons, this might explain selective routing. Indeed, when two visual stimuli induce two gamma rhythms in V1, only the gamma induced by the attended stimulus entrains gamma in V4. Here, we modelled induced responses with a Dynamic Causal Model for Cross-Spectral Densities and found that selective entrainment can be explained by attentional modulation of intrinsic V1 connections. Specifically, local inhibition was decreased in the granular input layer and increased in the supragranular output layer of the V1 circuit that processed the attended stimulus. Thus, presynaptic attentional influences and ensuing entrainment were sufficient to mediate selective routing.


Reducing urinary catheter use: a protocol for a mixed methods evaluation of an electronic reminder system in hospitalised patients in Australia.
Despite advances in infection prevention and control, catheter-associated urinary tract infections (CAUTIs) are common and remain problematic. Prolonged urinary catheterisation is the main risk factor for development of CAUTIs; hence, interventions that target early catheter removal warrant investigation. The study's objectives are to examine the efficacy of an electronic reminder system, the CATH TAG, in reducing urinary catheter use (device utilisation ratio) and to determine the effect of the CATH TAG on nurses' ability to deliver patient care. This study uses a mixed methods approach in which both quantitative and qualitative data will be collected. A stepped wedge randomised controlled design in which wards provide before and after observations will be undertaken in one large Australian hospital over 24 weeks. The intervention is the use of the CATH TAG. Eligible hospital wards will receive the intervention and act as their own control, with analysis undertaken of the change within each ward using data collected in control and intervention periods. An online survey will be administered to nurses on study completion, and a focus group for nurses will be conducted 2 months after study completion. The primary outcomes are the urinary catheter device utilisation ratio and perceptions of nurses about ease of use of the CATH TAG. Secondary outcomes include a reduced number of cases of catheter-associated asymptomatic bacteriuria, a reduced number of urinary catheters inserted per 100 patient admissions, perceptions of nurses regarding effectiveness of the CATH TAG, changes in ownership/interest by patients in catheter management, as well as possible barriers to successful implementation of the CATH TAG. Approval has been obtained from the Human Research Ethics Committees of Avondale College of Higher Education (2017:15) and Queensland Health (HREC17QTHS19). Results will be disseminated via peer-reviewed journals and conference presentations. ACTRN12617001191381 (Pre-results).


Retrospective analysis of the efficacy and tolerability of levetiracetam in patients with metastatic brain tumors.
Seizures are a common complication of metastatic brain tumors (MBT), affecting approximately 27-50% of all patients during the course of their illness. Treatment of tumor-induced seizures is often inadequate with traditional antiepileptic drugs (AED) due to a variety of factors, including activation of glutamatergic NMDA receptors, alterations of neuronal input pathways, and tumor growth. Levetiracetam (LEV) is a 2nd generation non-enzyme inducing AED with a novel mechanism of action, binding to neuronal synaptic vesicle protein SV2A, that has been previously shown to reduce seizure activity in patients with primary brain tumors. Due to its unique mechanism of action, it has been postulated that LEV may also be effective in controlling seizures from MBT. A retrospective chart review was performed of all Neuro-Oncology Center patients with MBT who had received LEV for seizure control. Thirteen patients were reviewed with a median age of 55.1 years (range: 34-70). Six patients had breast cancer, five had lung cancer, and two had melanoma. LEV was used as an add-on AED in seven patients (54%) and as monotherapy in six patients (46%), with a median dose of 1,000 mg/day (range: 500-3,000). The baseline median seizure frequency was one ictal event every other day. After the addition of LEV, the median seizure frequency was reduced to 0 per week. The seizure frequency was reduced to less than 50% of the pre-LEV baseline in 100% of patients (P=0.0002, Sign test), with 10 patients (77%; confidence interval: 46-95%) noting complete seizure control. The most common adverse event was somnolence and headache, noted in 3 of 13 patients (23%). LEV was very effective and well tolerated in MBT patients with seizures and should be considered for add-on therapy or as a substitute AED for monotherapy.


Detecting artificial anti-dengue IgM immune complexes using an enzyme-linked immunosorbent assay.
A variety of methods have been employed to detect viral immune complexes (IC) in clinical specimens. However, most techniques used were not antigen-specific. We developed a simple, specific double antibody sandwich technique to detect artificial anti-dengue (DEN) IgM immune complex (IgM-IC). Positive reactivity with IgM-ICs prepared with live DEN-1, -2, and -3 viruses was found to be related to IgM titers exceeding 1:20 and to the titer of the viruses. Most IgM-ICs prepared with live DEN-4 virus did not react. In contrast, IgM-ICs prepared with hemagglutination antigens, representing all 4 serotypes, reacted positively with amounts of antigens ranging from 2 to 8 units. These IgM-ICs were not type-specific.


2pi ambiguity-free optical distance measurement with subnanometer precision with a novel phase-crossing low-coherence interferometer.
We report a highly accurate phase-based technique for measuring arbitrarily long optical distance with subnanometer precision. The method employs a Michelson interferometer with a pair of harmonically related light sources, one cw and the other low coherence. By slightly detuning (~2 nm) the center wavelength of the low-coherence source between scans of the target sample, we can use the phase relationship between the heterodyne signals of the cw and the low-coherence light to measure the separation between reflecting interfaces with subnanometer precision. As this technique is completely free of 2pi ambiguity, an issue that plagues most phase-based techniques, it can be used to measure arbitrarily long optical distances without loss of precision. We demonstrate one application of this technique, the high-precision determination of the differential refractive index.


Evaluation of multiplexed hollow fiber flow field-flow fractionation for semi-preparative purposes.
A multiplexed hollow fiber flow field-flow fractionation (MxHF5) is introduced to increase throughput of an HF5 channel system for semi-preparative purposes. HF5, a cylindrical version of the flow field-flow fractionation (FlFFF) operated with a porous, hollow fiber membrane by controlling the ratio of radial and axial flow rates, is capable of fractionating proteins, cells, and macromolecules by size. An advantage of HF5 is its inexpensive channel construction, allowing for disposability that can reduce run-to-run carryover problems. MxHF5 constructed in this study was made with six parallel HF5 modules connected to seven-port manifolds for the semi-preparative scale separation of proteins or biological particles. For the evaluation of MxHF5 separation efficiency, protein standards were utilized to test peak recoveries, band broadening, and throughput. The assembly showed the possibility of handling up to 50 microg of proteins without incurring overloading. The developed channel was applied to demonstrate size sorting of lipoproteins for the future study of size dependent lipidomic and proteomic analysis. The current trial offers a unique advantage of scaling up HF5 separation without using wide-bore, hollow fibers which sacrifice separation speed.


Lactose binding to human galectin-7 (p53-induced gene 1) induces long-range effects through the protein resulting in increased dimer stability and evidence for positive cooperativity.
The product of p53-induced gene 1 is a member of the galectin family, i.e., galectin-7 (Gal-7). To move beyond structural data by X-ray diffraction, we initiated the study of the lectin by nuclear magnetic resonance (NMR) and circular dichroism spectroscopies, and molecular dynamics (MD) simulations. In concert, our results indicate that lactose binding to human Gal-7 induces long-range effects (minor conformational shifts and changes in structural dynamics) throughout the protein that result in stabilization of the dimer state, with evidence for positive cooperativity. Monte Carlo fits of (15)N-Gal-7 HSQC titrations with lactose using a two-site model yield K1 = 0.9 ± 0.6 × 10(3) M(-1) and K2 = 3.4 ± 0.8 × 10(3) M(-1). Ligand binding-induced stabilization of the Gal-7 dimer was supported by several lines of evidence: MD-based calculations of interaction energies between ligand-loaded and ligand-free states, gel filtration data and hetero-FRET spectroscopy that indicate a highly reduced tendency for dimer dissociation in the presence of lactose, CD-based thermal denaturation showing that the transition temperature of the lectin is significantly increased in the presence of lactose, and saturation transfer difference (STD) NMR using a molecular probe of the monomer state whose presence is diminished in the presence of lactose. MD simulations with the half-loaded ligand-bound state also provided insight into how allosteric signaling may occur. Overall, our results reveal long-range effects on Gal-7 structure and dynamics, which factor into entropic contributions to ligand binding and allow further comparisons with other members of the galectin family.


Efficient Flexible Fitting Refinement with Automatic Error Fixing for De Novo Structure Modeling from Cryo-EM Density Maps.
Structural modeling of proteins from cryo-electron microscopy (cryo-EM) density maps is one of the challenging issues in structural biology. De novo modeling combined with flexible fitting refinement (FFR) has been widely used to build a structure of new proteins. In de novo prediction, artificial conformations containing local structural errors such as chirality errors, cis peptide bonds, and ring penetrations are frequently generated and cannot be easily removed in the subsequent FFR. Moreover, refinement can be significantly suppressed due to the low mobility of atoms inside the protein. To overcome these problems, we propose an efficient scheme for FFR, in which the local structural errors are fixed first, followed by FFR using an iterative simulated annealing (SA) molecular dynamics protocol with the united atom (UA) model in an implicit solvent model; we call this scheme "SAUA-FFR". The best model is selected from multiple flexible fitting runs with various biasing force constants to reduce overfitting. We apply our scheme to the decoys obtained from MAINMAST and demonstrate an improvement of the best model of eight selected proteins in terms of the root-mean-square deviation, MolProbity score, and RWplus score compared to the original scheme of MAINMAST. Fixing the local structural errors can enhance the formation of secondary structures, and the UA model enables progressive refinement compared to the all-atom model owing to its high mobility in the implicit solvent. The SAUA-FFR scheme realizes efficient and accurate protein structure modeling from medium-resolution maps with less overfitting.


Imaging monitored loosening of dense fibrous tissues using high-intensity pulsed ultrasound.
Pulsed high-intensity focused ultrasound (HIFU) is proposed as a new alternative treatment for contracture of dense fibrous tissue. It is hypothesized that the pulsed-HIFU can release the contracted tissues by attenuating tensile stiffness along the fiber axis, and that the stiffness reduction can be quantitatively monitored by change of B-mode images. Fresh porcine tendons and ligaments were adapted to an ex vivo model and insonated with pulsed-HIFU for durations ranging from 5 to 30 min. The pulse length was 91 µs with a repetition frequency of 500 Hz, and the peak rarefactional pressure was 6.36 MPa. The corresponding average intensities were kept around 1606 W cm(-2) for ISPPA and 72.3 W cm(-2) for ISPTA. B-mode images of the tissues were acquired before and after pulsed-HIFU exposure, and the changes in speckle intensity and organization were analyzed. The tensile stiffness of the HIFU-exposed tissues along the longitudinal axis was examined using a stretching machine. Histology examinations were performed by optical and transmission electron microscopy. Pulsed-HIFU exposure significantly decreased the tensile stiffness of the ligaments and tendons. The intensity and organization of tissue speckles in the exposed region were also decreased. The speckle changes correlated well with the degree of stiffness alteration. Histology examinations revealed that pulsed-HIFU exposure probably damages tissues via a cavitation-mediated mechanism. Our results suggest that pulsed-HIFU with a low duty factor is a promising tool for developing new treatment strategies for orthopedic disorders.


Molecular cloning and characterization of the human CLOCK gene: expression in the suprachiasmatic nuclei.
The Clock gene is an essential regulator of circadian rhythms. It encodes a member of the basic helix-loop-helix/PER-ARNT-SIM family of transcription factors known to play a central role in the control of diverse cellular events. Previously we described the functional identification and molecular isolation of the Clock gene in the mouse, its interaction with the BMAL1 protein, and the role of this complex as a transcriptional activator in the circadian pacemaker. Here, we report the cloning, exon organization, chromosomal location, and mRNA expression of the human CLOCK gene. The coding sequence of human CLOCK extends for 2538 bp and is 89% identical to its mouse ortholog; its deduced amino acid sequence is 846 residues long and is 96% identical to mouse CLOCK. Radiation hybrid mapping localized human CLOCK to the long arm of human chromosome 4 (4q12). Direct sequencing of a genomic CLOCK clone indicated that the coding sequence of human CLOCK extends over 20 exons and that its intron/exon organization is identical to that of the mouse ortholog. Northern blot analysis indicated widespread expression of two major transcripts of 8 and 10 kb, and in situ hybridization of human brain tissue revealed elevated expression of CLOCK mRNA in the suprachiasmatic nuclei, the locus of circadian control in mammals, and in the cerebellum. Comparison of cDNA clones revealed two single nucleotide polymorphisms in noncoding sequence flanking the CLOCK open reading frame. The central role of Clock in the organization of circadian rhythms suggests that it will be a useful candidate gene for genetic analyses of disorders associated with dysfunction of the circadian system.


Amelioration of prediabetes-induced changes of dendritic structural plasticity.
Accumulating evidence suggests that the diabetes-induced cognitive dysfunction can be alleviated when exposed to the enriched environment. However, the impact of the changes of the hippocampal plasticity on the cognitive decline and the possible effect of an enriched environment in prediabetes are still not clearly documented. To explore the effect of enriched environment for prediabetes-induced changes of dendritic structural plasticity in hippocampus pyramidal and cognitive deficits, the praxiology experiments for evaluating of anxiety, spatial learning and memory of prediabetic Wistar were performed, and then the dendritic spine density was assessed in the hippocampal CA1 pyramidal neuronal region. The prediabetic rats demonstrated a hyper-anxiety like behavior and significantly decreased spatial learning abilities and memory deficits. Exposing prediabetic rats to an enriched environment appeared to significantly mitigate the above changes in a time-dependent manner. The enriched environment also restored the density of the hippocampal dendritic spine which was significantly reduced in prediabetic rats. We found that the enriched environment was beneficial in overcoming the prediabetes-induced cognitive disorders and diminished dendritic plasticity of hippocampus pyramidal.


Interfacially driven transport theory: a way to unify Marangoni and osmotic flows.
We show that the solvent behaviour in both diffusio-osmosis and Marangoni flow can be derived from a simple model of colloid-interface interactions. We demonstrate that the direction of the flow is regulated by a single value of the attractive parameter covering the purely repulsive and attractive-repulsive interaction cases. The proposed universality between diffusio-osmosis and Marangoni flow is extended further to include diffusio-phoresis. In particular, an object immersed to a colloidal solution moves towards the low concentration of the colloidal particles in the case of colloid-interface repulsion and towards the high concentration of the colloidal particles in the case of colloid-interface attraction. The approach combines the methods of fluid dynamics, molecular physics and transport phenomena and provides a tractable explanation of how the colloid-interface interactions affect the momentum balance and the transport phenomena (interfacially driven transport).


[Eyelid hygiene in preparing dry eye patients for cataract surgical treatment].
To evaluate the effectiveness of complex treatment, including eyelid hygiene, on the condition of ocular surface in dry eye patients before cataract surgery. The study included 56 female age-related cataract patients (64±4.3 years old) with dry eye caused by Meibomian gland dysfunction (MGD). The main group patients (n=28) underwent eyelid hygiene therapy (2 times a day, 1 month) that involved Blefarogel cleansing, Blefarolotion, Blefarogel-1. The main and control (n=28) group patients performed (during the month): «fat-water» type nanoemulsion instillation (3 times a day); preservative free vitamin-A application (at night). Methods included dry eye symptoms assessment (OSDI scale); TBUT; Shirmer-1 test; lissamine green vital staining with xerosis indicator calculation (van Bijsterveld scale); "lid wipers" symptom evaluation; Norn compression test. Statistical evaluation included: mean and standard deviation calculation (M±s); significance of differences assessment (Mann-Whitney U-test; Wilcoxon's T-test). The main group patients (artificial tear, keratoprotection, eyelid hygiene) showed a statistically significant more pronounced decrease in OSDI (from 33.2±3.1 points to 15.2±1.8 points) and a significantly more pronounced increase in TBUT (from 4.8±0.5 s to 6.9±0.4 s) than in control group patients (artificial tear, keratoprotection; respectively, from 32.7±3.2 to 20.8±3.1 points and from 4.85±0.5 to 5.8±0.4 s; p<0.05). Apparently, this was associated with a significantly more pronounced relief of the MGD signs in main group patients (from 2.1±0.1 to 1.1±0.2 points; p<0.05) as a result of eyelid hygiene. In the control group, the dynamics of MGD severity (from 2.08±0.3 to 2.11±0.5 points) was statistically insignificant. Eyelid hygiene (based on Blefarogel cleansing, Blefarolotion, Blefarogel-1) increases the effectiveness of dry eye treatment in MGD patients, compared with isolated tear substitution and keratoprotection.


A mechanistic cohort study evaluating cognitive impairment in women treated for breast cancer.
Some women report cognitive impairment after adjuvant chemotherapy (CTh) for breast cancer. Here we explore cognitive function, and underlying mechanisms with blood tests and functional magnetic resonance imaging (fMRI). Women treated for early breast cancer were recruited to three groups based on self-reported cognitive symptoms (CS) using FACT-Cog scores. CTh + CS+ (n = 44) had received chemotherapy and self-reported cognitive symptoms; CTh + CS- (n = 52) had chemotherapy but did not report cognitive problems; CTh- (n = 30) had not received chemotherapy. Clinical and computer-based neuropsychological tests were performed. Blood tests included 10 cytokines, sex hormones, coagulation factors, and apolipoprotein-E genotype. fMRI (n = 101) was performed while subjects performed an n-back memory task. Participants had median age 50 (range: 29-60) years and were a median of 17 months post-diagnosis. On clinical neuropsychological tests 19% had cognitive impairment using Global Deficit Score, and 36% using International Cancer and Cognition Task Force criteria with no significant differences in cognitive impairment rates between groups. CTh + CS+ had significantly more fatigue, anxiety/depression and poorer quality-of-life than other groups. There was no association between FACT-Cog and neuropsychological scores. There were significant differences in frontal and parietal regions on fMRI scans: CTh- showed hyperactivation compared to chemotherapy-treated groups, CTh + CS+ had more frontal activation than CTh + CS-. Elevated IL-1, IL-2 were associated weakly and IL-8 more strongly with neuropsychological impairment (rho > 0.20). There were no differences in global cognitive impairment between groups. Cognitive symptoms were associated with fatigue and anxiety/depression, but not with objective cognitive impairment. fMRI scans differed among the three groups.