Long-term modulation of presynaptic 5-HT-output: experimentally induced changes in cortical 5-HT-transporter density, tryptophan hydroxylase content and 5-HT innervation density.
Whereas experimentally induced long-term changes of postsynaptic mechanisms of 5-HT neurotransmission have been studied in great detail, much less is currently known about the effects of certain treatments on the presynaptic components governing the output of 5-HT in individual brain regions. This contribution summarizes the results of a series of experiments on the influence of different physiologic and pharmacologic manipulations on three different parameters of 5-HT presynapses, 5-HT transporter density, tryptophan hydroxylase content, and serotonin level in the rat frontal cortex. The combined measurement of several parameters of 5-HT presynapses allows to differentiate between treatments which exclusively affect the density of 5-HT transporters (long-term food restriction), which exclusively affect the level of tryptophan hydroxylase apoenzyme (imipramine treatment of olfactory bulbectomized rats) or which cause a parallel increase (bulbectomy, chronic administration of tranylcypramine to rats with chemical lesions of their cortical 5-HT innervation) or a parallel decrease (administration of p-chloroamphetamine) of both parameters, indicating treatment-induced changes in the density of 5-HT presynapses in the frontal cortex. Each of these changes may lead to an altered output of serotonergic afferences, and may therefore act to either potentiate or to attenuate the impact of serotonin-mediated effects on the activity of local networks located in a certain brain region.


Matriptase proteolytically activates influenza virus and promotes multicycle replication in the human airway epithelium.
Influenza viruses do not encode any proteases and must rely on host proteases for the proteolytic activation of their surface hemagglutinin proteins in order to fuse with the infected host cells. Recent progress in the understanding of human proteases responsible for influenza virus hemagglutinin activation has led to the identification of members of the type II transmembrane serine proteases TMPRSS2 and TMPRSS4 and human airway trypsin-like protease; however, none has proved to be the sole enzyme responsible for hemagglutinin cleavage. In this study, we identify and characterize matriptase as an influenza virus-activating protease capable of supporting multicycle viral replication in the human respiratory epithelium. Using confocal microscopy, we found matriptase to colocalize with hemagglutinin at the apical surface of human epithelial cells and within endosomes, and we showed that the soluble form of the protease was able to specifically cleave hemagglutinins from H1 virus, but not from H2 and H3 viruses, in a broad pH range. We showed that small interfering RNA (siRNA) knockdown of matriptase in human bronchial epithelial cells significantly blocked influenza virus replication in these cells. Lastly, we provide a selective, slow, tight-binding inhibitor of matriptase that significantly reduces viral replication (by 1.5 log) of H1N1 influenza virus, including the 2009 pandemic virus. Our study establishes a three-pronged model for the action of matriptase: activation of incoming viruses in the extracellular space in its shed form, upon viral attachment or exit in its membrane-bound and/or shed forms at the apical surface of epithelial cells, and within endosomes by its membrane-bound form where viral fusion takes place.


Adolescents and violence: "adults are cruel: "they just beat, beat, beat!".
This paper examines the nature of violence as experienced by adolescent African children between the ages of 10-14 years in the South African township of New Crossroads near Cape Town. The longitudinal study focuses on children from a wide variety of family structures and dynamics in an attempt to examine detailed factors shaping each adolescent's development. The study moves from the premise that the impact of violence on individuals is complex, reflecting the interaction between the violent act on the one hand, and coping strategies adopted by the victims on the other. The children identified three spheres as arenas in which violent interactions occur: home, school and street. It was found that not only have the children devised strategies for dealing with the violence and learned how to negotiate various relationships with adults and peers but also that beyond the violence around them they do have some aspirations.


Adaptive distributed outlier detection for WSNs.
The paradigm of pervasive computing is gaining more and more attention nowadays, thanks to the possibility of obtaining precise and continuous monitoring. Ease of deployment and adaptivity are typically implemented by adopting autonomous and cooperative sensory devices; however, for such systems to be of any practical use, reliability and fault tolerance must be guaranteed, for instance by detecting corrupted readings amidst the huge amount of gathered sensory data. This paper proposes an adaptive distributed Bayesian approach for detecting outliers in data collected by a wireless sensor network; our algorithm aims at optimizing classification accuracy, time complexity and communication complexity, and also considering externally imposed constraints on such conflicting goals. The performed experimental evaluation showed that our approach is able to improve the considered metrics for latency and energy consumption, with limited impact on classification accuracy.


The GIY-YIG Type Endonuclease Ankyrin Repeat and LEM Domain-Containing Protein 1 (ANKLE1) Is Dispensable for Mouse Hematopoiesis.
Ankyrin repeat and LEM-domain containing protein 1 (ANKLE1) is a GIY-YIG endonuclease with unknown functions, mainly expressed in mouse hematopoietic tissues. To test its potential role in hematopoiesis we generated Ankle1-deficient mice. Ankle1Δ/Δ mice are viable without any detectable phenotype in hematopoiesis. Neither hematopoietic progenitor cells, myeloid and lymphoid progenitors, nor B and T cell development in bone marrow, spleen and thymus, are affected in Ankle1Δ/Δ-mice. Similarly embryonic stress erythropoiesis in liver and adult erythropoiesis in bone marrow and spleen appear normal. To test whether ANKLE1, like the only other known GIY-YIG endonuclease in mammals, SLX1, may contribute to Holliday junction resolution during DNA repair, Ankle1-deficient cells were exposed to various DNA-damage inducing agents. However, lack of Ankle1 did not affect cell viability and, unlike depletion of Slx1, Ankle1-deficiency did not increase sister chromatid exchange in Bloom helicase-depleted cells. Altogether, we show that lack of Ankle1 does neither affect mouse hematopoiesis nor DNA damage repair in mouse embryonic fibroblasts, indicating a redundant or non-essential function of ANKLE1 in mouse.


Conformational change in a single molecular species, beta3, of beta-conglycinin in acidic ethanol solution.
Several physicochemical experiments were done to obtain further information on the conformational changes occurring in beta-conglycinin in acidic-ethanol solution, using a single molecular species of this protein, beta3. By far-UV circular dichroism (CD), a transition from beta-sheet to alpha-helical structure was observed upon addition of acidic-ethanol, and the alpha-helix content was found to reach 76% in 70% ethanol (pH 2). From analyses of near-UV CD and difference absorption spectra, it was found that the tertiary structure of the beta3 species was significantly altered at ethanol concentrations between 10 and 20%. The profiles of binding of 1-anilinonaphthalene-8-sulfonic acid to the beta3 species during acidic-ethanol denaturation were indicative of the existence of intermediate conformers in the molten globule-like denaturation state. By measuring Fourier transform infrared spectra and estimating the Stokes radius by dynamic light scattering, the beta3 molecules were found to aggregate with an increase in ethanol concentration.


An Atraumatic Sacral Fracture with Lumbosacral Radiculopathy Complicating the Early Postpartum Period: A Case Report.
BACKGROUND Sacral stress fractures are rare complications of pregnancy and the early postpartum. Of these, few present with lumbosacral radiculopathy. We report the first Australian case of a young multiparous woman who sustained an atraumatic, fatigue sacral fracture with associated radiculopathy. We highlight the diagnostic process and chronic management of this case, particularly in relation to a future pregnancy. CASE REPORT A 26-year-old multiparous Caucasian female presented with worsening lumbosacral back pain and radicular symptoms following the rapid and spontaneous vaginal delivery of her second infant. Her pregnancy was unremarkable and she had no personal risk factors for osteoporosis. A magnetic resonance imaging (MRI) scan confirmed the diagnosis of a right S1 vertebral fracture. Bone densitometry and fasting bone metabolic testing excluded pregnancy-associated osteoporosis. She was managed conservatively with intermittent bed rest, regular physiotherapy and multimodal analgesia. During a future pregnancy, she experienced a severe exacerbation of her lumbosacral radiculopathy requiring hospital admission, up-titration of her analgesia and a right S1 epidural injection. She subsequently underwent an elective caesarean section and has since benefitted from regular hydrotherapy. CONCLUSIONS Lumbosacral radiculopathy in the absence of trauma during pregnancy or the early postpartum should prompt consideration of an underlying atraumatic, fatigue sacral fracture. Such fractures may result from the abnormal biomechanical loading of the sacrum during rapid vaginal deliveries and are most effectively diagnosed by MRI. Conservative management strategies involving physiotherapy and multimodal analgesia are recommended. Future pregnancies may exacerbate radicular symptoms. Such patients may subsequently benefit from elective caesarean section deliveries and hydrotherapy.


Mechanism of interaction between ascorbic acid and soil iron-containing minerals for peroxydisulfate activation and organophosphorus flame retardant degradation.
Soil constituents may play an important role in peroxydisulfate (PDS)-based oxidation of organic contaminants in soil. Iron-containing minerals (Fe-minerals) have been found to promote PDS activation for organics degradation. Our study found that ascorbic acid (H2A) could enhance PDS activation by soil Fe-minerals for triphenyl phosphate (TPHP) degradation. Determination and characterization analyses of Fe fractions showed that H2A could induce the reductive dissolution of solid Fe-minerals and the increasing of oxygen vacancies/hydroxyl groups content on Fe-minerals surface. The increasing of divalent Fe (Fe(Ⅱ)) accelerated PDS activation to generate reactive oxygen species (ROS). Electron paramagnetic resonance (EPR) and quenching studies showed that sulfate radicals (SO4•-) and hydroxyl radicals (HO•) contributed significantly to TPHP degradation. The composition and content of Fe-minerals and soil organic matter (SOM) markedly influenced ROS transformations. Surface-bond and structural Fe played the main role in the production of Fe(Ⅱ) in reaction system. The high-concentration SOM could result in ROS consumption and degradation inhibition. Density functional theory (DFT) studies revealed that H2A is preferentially adsorbed at α-Fe2O3(012) surface through Fe-O-C bridges rather than hydrogen bonds. After absorption, H atoms on H2A may further is migrated to adjacent O atoms on the α-Fe2O3(012) surface. With the transformation of H atoms to the α-Fe2O3(012) surface, the Fe-O-C bridge is broken and one electron is transferred from the O to Fe atom, inducing the reduction of trivalent Fe (Fe(III)) atom. MS/MS2 analysis, HPLC analysis, and toxicity assessment demonstrated that TPHP was transformed to less toxic 4-hydroxyphenyl diphenyl phosphate (OH-TPHP), diphenyl hydrogen phosphate (DPHP), and phenyl phosphate (PHP) through phenol-cleavage and hydroxylation processes, and even be mineralized in reaction system.


Subtidal gastropods consume sulfur-oxidizing bacteria: evidence from coastal hydrothermal vents.
The black abalone (Haliotis cracherodii), a commercially important shallow-water gastropod common off White Point, Southern California, is found frequently at subtidal hydrothermal vents within mats of filamentous sulfur-oxidizing bacteria. Foraging vent abalones actively consume the bacteria and confine their nightly feeding forays to bacterial mats surrounding the vents. The growth of abalones consuming the sulfur bacteria exceeds that of control individuals consuming microalgae and is comparable to reported growth rates of abalones consuming macroalgae. Thus, off White Point, the black abalone may derive a portion of its nutrition from the subsidy of geothermal energy.


Role of mTOR in podocyte function and diabetic nephropathy in humans and mice.
Chronic glomerular diseases, associated with renal failure and cardiovascular morbidity, represent a major health issue. However, they remain poorly understood. Here we have reported that tightly controlled mTOR activity was crucial to maintaining glomerular podocyte function, while dysregulation of mTOR facilitated glomerular diseases. Genetic deletion of mTOR complex 1 (mTORC1) in mouse podocytes induced proteinuria and progressive glomerulosclerosis. Furthermore, simultaneous deletion of both mTORC1 and mTORC2 from mouse podocytes aggravated the glomerular lesions, revealing the importance of both mTOR complexes for podocyte homeostasis. In contrast, increased mTOR activity accompanied human diabetic nephropathy, characterized by early glomerular hypertrophy and hyperfiltration. Curtailing mTORC1 signaling in mice by genetically reducing mTORC1 copy number in podocytes prevented glomerulosclerosis and significantly ameliorated the progression of glomerular disease in diabetic nephropathy. These results demonstrate the requirement for tightly balanced mTOR activity in podocyte homeostasis and suggest that mTOR inhibition can protect podocytes and prevent progressive diabetic nephropathy.


Circadian patterns of plasma cortisol, 17-hydroxyprogesterone, and testosterone in congenital adrenal hyperplasia.
In 11 children aged between 2 and 17 years with (nonsalt-losing) congenital adrenal hyperplasia (21-hydroxylase deficiency) blood was drawn at 90-minute intervals during a 24-hour period and levels of 17-hydroxyprogesterone, testosterone, and cortisol were measured. Levels of 17-ketosteroids and pregnanetriol were measured too in 24-hour urine samples. These measurements were taken under different regimens of treatment and after interruption of treatment. Cortisol level rose and fell rapidly after administered corticosteroid, and reached unphysiologically high levels. Testosterone levels showed pronounced variations but stayed in the normal range for most of the time even in untreated patients; thus testosterone provides a poor control parameter. Levels of 17-hydroxyprogesterone showed extreme fluctuations and very high peak levels in untreated patients; standard treatment with two or three daily doses of corticosteroids did not prevent a pronounced rise in its level after midnight. After the first morning dose of hydrocortisone a very steep fall was observed. The 24-hour pregnanetriol excretion correlated well with the corresponding total integrated 17-hydroxyprogesterone area. It is concluded that single 17-hydroxyprogesterone values are unlikely to give adequate information about the quality of treatment.


Bioinformatic Mapping of Opine-Like Zincophore Biosynthesis in Bacteria.
Zinc is an essential nutrient in biological systems due to its structural or catalytic requirement in proteins involved in diverse cellular processes. To meet this cellular demand, microbes must acquire sufficient zinc from their environment. However, many environments have low zinc availability. One of the mechanisms used by bacteria to acquire zinc is through the production of small molecules known as zincophores. Similar to bacterial siderophores used for iron uptake, zincophores are synthesized by the bacterium and exported and then reimported as zincophore-zinc complexes. Thus far, only four zincophores have been described, including two from the human pathogens Staphylococcus aureus and Pseudomonas aeruginosa, in which they play a critical role in zinc acquisition during infection, and one in a soil bacterium. To determine what other microbes may produce zincophores, we used bioinformatic analyses to identify new zincophore biosynthetic gene clusters (BGCs) and predict the diversity of molecules synthesized. Genome neighborhood network analysis identified approximately 250 unique zincophore-producing species from actinobacteria, firmicutes, proteobacteria, and fusobacteria. This indicates that zincophores are produced by diverse bacteria that inhabit a broad range of ecological niches. Many of the BGCs likely produce characterized zincophores, based on similarity to the characterized systems. However, this analysis also identified numerous BGCs that, based on the colocalization of additional modifying enzymes and sequence divergence of the biosynthetic enzymes, are likely to produce unique zincophores. Collectively, these findings provide a comprehensive understanding of the zincophore biosynthetic landscape that will be invaluable for future research on these important small molecules.IMPORTANCE Bacteria must acquire essential nutrients, including zinc, from their environment. For bacterial pathogens, this necessitates overcoming the host metal-withholding response known as nutritional immunity. A novel type of zinc uptake mechanism that involves the bacterial production of a small zinc-scavenging molecule was recently described in the human pathogens Staphylococcus aureus, Pseudomonas aeruginosa, and Yersinia pestis, as well as the soil-associated bacterium Paenibacillus mucilaginosus This suggests that zincophores may be important for zinc acquisition in diverse environments. In this study, we sought to identify other zincophore-producing bacteria using bioinformatics. We identified almost 250 unique zincophore-producing species, including human and animal pathogens, as well as isolates from soil, rhizosphere, plant, and marine habitats. Crucially, we observed diversity at the amino acid and gene organization levels, suggesting that many of these species are producing unique zincophores. Together, our findings highlight the importance of zincophores for a broad array of bacteria living in diverse environments.


Effects of cyclooxygenase inhibition on insulin release and pancreatic islet blood flow in rats.
To examine the effects of inhibition of cyclooxygenase (COX) on islet hormone secretion in vitro and on pancreatic islet blood flow in vivo. Insulin release was measured in a static incubation system of islets isolated from Wistar-F rats after inhibition of COX-1 and COX-2 with SC 560 (COX-1), FR 122047 (COX-1), rofecoxib (COX-2), or indomethacin (both COX-1 and COX-2). In other rats organ blood flow values were measured with a microsphere technique during both normo- and hyperglycemia after administration of these enzyme inhibitors. Serum insulin values were lower after pretreatment with a COX-1 inhibitor or a non-selective COX inhibitor in both control and glucose-injected rats in vivo, whereas COX-2 inhibition had no such effects. However, inhibition of COX had only minor effects on insulin release in vitro. Inhibition of COX affected neither total pancreatic nor islet blood flow in normoglycemic rats. Hyperglycemia caused an increase in both these flow values and in the duodenum. The increase in total pancreatic and duodenal blood flow was prevented by inhibition of COX-2 or non-selective COX inhibition. However, no effects on islet blood flow were seen after COX inhibition. Inhibition of COX affects insulin release and blood glucose concentrations in vivo. However, COX inhibition has only minor effects on pancreatic islet blood flow, but prevents the glucose-induced increase in total pancreatic blood flow.


RNA-Seq Reveals that Light and Darkness Are Different Stimuli in Freshwater Heterotrophic Actinobacteria.
Light is a ubiquitous source of both energy and information in surface environments, and regulates gene expression not only in photosynthetic microorganisms, but in a broad range of photoheterotrophic and heterotrophic microbes as well. Actinobacteria are keystone species in surface freshwater environments, where the ability to sense light could allow them to coordinate periods of nutrient uptake and metabolic activity with primary production. The model freshwater Actinobacteria Rhodoluna (R.) lacicola strain MWH-Ta8 and Aurantimicrobium (A.) photophilum strain MWH-Mo1 grow faster in the light than in the dark, but do not use light energy to support growth. Here, we characterize transcription throughout a light-dark cycle in R. lacicola and A. photophilum. In both species, some genes encoding carbohydrate metabolism and storage are upregulated in the light. However, expression of genes of the TCA cycle is only coordinated with light availability in R. lacicola. In fact, the majority of genes that respond to light and darkness in these two species are different, even though their light-responsive phenotypes are similar. The ability to respond to light and darkness may be widespread in freshwater Actinobacteria, but the genetic networks controlled by these two stimuli may vary significantly.


Oscillating neuro-capillary coupling during cortical spreading depression as observed by tracking of FITC-labeled RBCs in single capillaries.
Coupling between capillary red blood cell (RBC) movements and neuronal dysfunction during cortical spreading depression (CSD) was examined in rats by employing a high-speed camera laser-scanning confocal fluorescence microscope system in conjunction with our Matlab domain software (KEIO-IS2). Following microinjection of K(+) onto the surface of the brain, changes in electroencephalogram (EEG), DC potential and tissue optical density were all compatible with the occurrence of a transient spreading neuronal depression. RBC flow in single capillaries was not stationary. Unpredictable redistribution of RBCs at branches of capillaries was commonly observed, even though no change in diameter was apparent at the reported site of the capillary sphincter and no change of arteriolar-venule pressure difference was detected. There appeared to be a slow morphological change of astroglial endfeet. When local neurons were stunned transiently by K(+) injection, the velocity and oscillation frequency of RBCs flowing in nearby capillaries started to decrease. The flow in such capillaries was rectified, losing oscillatory components. Sluggish floating movements of RBCs in pertinent capillaries were visualized, with occasional full stops. When CSD subsided, RBC movements recovered to the original state. We postulate that neuronal depolarization blocks oscillatory signaling to local capillaries via low-shear plasma viscosity increases in the capillary channels, and a complex interaction between the RBC surface and the buffy coat on the capillary wall surface increases the capillary flow resistance. Then, when CSD subsides and oscillatory neuronal function is recovered, the normal physiological conditions are restored.


Resting-state glutamatergic neurotransmission is related to the peak latency of the auditory mismatch negativity (MMN) for duration deviants: An (1)H-MRS-EEG study.
Mismatch negativity (MMN), an ERP elicited by a deviant stimulus in a train of standard stimuli, has been suggested to be associated to glutamatergic neurotransmission, mediated by glutamatergic NMDA receptors. In this study, we examined the relationship between interindividual variation of (1)H-MRS-measured glutamate+glutamine (Glx) in the superior temporal gyrus and MMN for duration and frequency deviants in 19 healthy young adults (9 male). We found a significant relationship between the peak latency of the duration-MMN peak and creatine-scaled Glx (p = .0003, η(2) = .43), with increased Glx level being associated to earlier peak of the duration-MMN (r = -.63). In contrast, the amplitude of the duration-MMN was not related to Glx. There was no significant relationship between Glx and the frequency-MMN. The present study is the first to demonstrate that interindividual variation in the glutamatergic neurotransmission affects the MMN response in healthy individuals.


The importance of management information systems in a managed care environment.
Keys to successful information systems for home care providers are planning and control. With managed care's emphasis on data, agencies need to have information systems that can handle the demands managed care puts on agencies today--planning before hurrying to install a system will ensure control as the managed care contracts add up.


Autophagic bulk sequestration of cytosolic cargo is independent of LC3, but requires GABARAPs.
LC3, a mammalian homologue of yeast Atg8, is assumed to play an important part in bulk sequestration and degradation of cytoplasm (macroautophagy), and is widely used as an indicator of this process. To critically examine its role, we followed the autophagic flux of LC3 in rat hepatocytes during conditions of maximal macroautophagic activity (amino acid depletion), combined with analyses of macroautophagic cargo sequestration, measured as transfer of the cytosolic protein lactate dehydrogenase (LDH) to sedimentable organelles. To accurately determine LC3 turnover we developed a quantitative immunoblotting procedure that corrects for differential immunoreactivity of cytosolic and membrane-associated LC3 forms, and we included cycloheximide to block influx of newly synthesized LC3. As expected, LC3 was initially degraded by the autophagic-lysosomal pathway, but, surprisingly, autophagic LC3-flux ceased after ~2h. In contrast, macroautophagic cargo flux was well maintained, and density gradient analysis showed that sequestered LDH partly accumulated in LC3-free autophagic vacuoles. Hepatocytic macroautophagy could thus proceed independently of LC3. Silencing of either of the two mammalian Atg8 subfamilies in LNCaP prostate cancer cells exposed to macroautophagy-inducing conditions (starvation or the mTOR-inhibitor Torin1) confirmed that macroautophagic sequestration did not require the LC3 subfamily, but, intriguingly, we found the GABARAP subfamily to be essential.


Lycopene and apo-10'-lycopenoic acid have differential mechanisms of protection against hepatic steatosis in β-carotene-9',10'-oxygenase knockout male mice.
Nonalcoholic fatty liver disease is positively associated with obesity and cardiovascular disease risk. Apo-10'-lycopenoic acid (APO10LA), a potential oxidation product of apo-10'-lycopenal that is generated endogenously by β-carotene-9',10'-oxygenase (BCO2) cleavage of lycopene, inhibited hepatic steatosis in BCO2-expressing mice. The present study evaluated lycopene and APO10LA effects on hepatic steatosis in mice without BCO2 expression. Male and female BCO2-knockout (BCO2-KO) mice were fed a high saturated fat diet (HSFD) with or without APO10LA (10 mg/kg diet) or lycopene (100 mg/kg diet) for 12 wk. Lycopene or APO10LA supplementation reduced hepatic steatosis incidence (78% and 72%, respectively) and severity in BCO2-KO male mice. Female mice did not develop steatosis, had greater hepatic total cholesterol (3.06 vs. 2.31 mg/g tissue) and cholesteryl ester (1.58 vs. 0.86 mg/g tissue), but had lower plasma triglyceride (TG) (229 vs. 282 mg/dL) and cholesterol (97.1 vs. 119 mg/dL) than male mice. APO10LA-mitigated steatosis in males was associated with reduced hepatic total cholesterol (18%) and activated sirtuin 1 signaling, which resulted in reduced fatty acids (FAs) and TG synthesis markers [stearoyl-coenzyme A (CoA) desaturase protein, 71%; acetyl-CoA carboxylase phosphorylation, 79%; AMP-activated protein kinase phosphorylation, 67%], and elevated cholesterol efflux genes (cytochrome P450 family 7A1, 65%; ATP-binding cassette transporter G5/8, 11%). These APO10LA-mediated effects were not mimicked by lycopene supplementation. Intriguingly, steatosis inhibition by lycopene induced peroxisome proliferator-activated receptor (PPAR)α- and PPARγ-related genes in mesenteric adipose tissue (MAT) that increases mitochondrial uncoupling [cell death-inducing DNA fragmentation factor, α subunit-like effector a, 55%; PR domain-containing 16, 47%; uncoupling protein 3 (Ucp3), 55%], FA β-oxidation (PPARα, 53%; very long chain acyl-CoA dehydrogenase, 38%), and uptake (FA transport protein 4, 29%; lipoprotein lipase 43%). Expressions of 10 MAT PPAR-related genes were inversely correlated with steatosis score, suggesting that lycopene reduced steatosis by increasing MAT FA utilization. Our data suggest that lycopene and APO10LA inhibit HSFD-induced steatosis in BCO2-KO male mice through differential mechanisms. Sex disparity of BCO2-KO mice was observed in the outcomes of HSFD-induced liver steatosis and plasma lipids.


[Changes in the ligand affinity of benzodiazepine receptors in the presence of 4,5,6,7-tetrahydroisoxazolo(5,4-c)pyridin-3-ol (THIP)].
4,5,6,7-tetrahydroisoxazolo-(5,4-c-)pyridin-3-ol (THIP) in concentrations of 100-500 microM inhibited 3H-flunitrazepam (3H FNZ) binding to intact membranes at 0 degrees C in 25 mM TRIS-HCl buffer. The inhibition of 3H FNZ binding was due to the decrease in the affinity and Bmax of 3H FNZ binding. The affinity for benzodiazepines (e.g. BZ-receptor agonists) was reduced in the presence of THIP, whereas the affinity for BZ-receptor antagonists was unchanged. The value of THIP-induced shift was dependent on the molarity of TRIS-HCl buffer and the concentration of THIP. The results obtained suggest that THIP-induced shifts have a predictive value in the determination of pharmacological efficacy of BZ-receptor ligands.


Binding of adenosine to the crude plasma membrane fraction isolated from dog coronary and carotid arteries.
In an attempt to elucidate the mechanism of action of adenosine, specific adenosine binding was determined by using plasma membrane rich microsomal fraction of dog coronary and carotid arteries. The binding of adenosine to both coronary and carotid preparations was temperature as well as pH dependent and was inhibited by aminophylline and adenosine nucleotides. Scatchard analysis of the saturable binding data on carotid arteries exhibited a single species of binding sites with a Kd value of 1.34 x 10(-6) M and binding capacity of 140 pmol/mg of protein. The first order dissociation rate constant of adenosine binding to carotid preparation was 0.0247 min-1. The nonlinearity of equilibrum binding data (Scatched Plot) and dissociation curve of binding to coronary arteries suggested the presence of more than one population of binding sites. The data suggest that the binding sites located in the microsomal fraction of coronary and carotid arteries possess the basic characteristics of the receptors which might be involved in the vasodilatory action of adenosine.


Performance of soluble Klotho assays in clinical samples of kidney disease.
Soluble Klotho has multiple systemic salutary effects. In animals, both acute and chronic kidney disease models display systemic Klotho deficiency. As such, there is considerable interest in investigating soluble Klotho as a biomarker in patients with different types and severity of kidney diseases. Unfortunately, there remains uncertainty regarding the best method to measure soluble Klotho in human serum samples. Using human serum samples obtained from several clinical cohorts with a wide range of kidney function, we measured soluble Klotho using a commercial enzyme-linked immunosorbent assay (ELISA) as well as with an immunoprecipitation-immunoblot (IP-IB) assay utilizing a synthetic antibody with high affinity and specificity for Klotho. Recovery of spiking with a known amount of exogenous Klotho was tested. A subset of samples was analyzed with and without the addition of a protease inhibitor cocktail at the time of collection or after the first freeze-thaw cycle to determine if these maneuvers influenced performance. The IP-IB assay was superior to the ELISA at recovery of exogenous Klotho (81-115% versus 60-81%) across the spectrum of kidney function. Klotho measurements by IP-IB were highly correlated with estimated glomerular filtration rate (eGFR) (R = 0.80, P < 0.001) in comparison with the commercial ELISA, which exhibited minimal correlation with eGFR (R = 0.18, P = 0.12). Use of a protease inhibitor cocktail neither improved nor impaired performance of the IP-IB assay; however, subsequent freeze-thaw cycle resulted in a significant reduction in Klotho recovery and dissipated the correlation between Klotho levels and eGFR. With the ELISA, the use of protease inhibitor cocktail resulted in an increase in intrasubject variability. The IP-IB assay is preferable to the commercial ELISA to measure soluble Klotho concentrations in never-thawed serum samples of humans with varying severity of kidney disease. However, due to the labor-intensive nature of the IP-IB assay, further research is needed to secure an assay suitable for high-throughput work.


Healthy Eating for Elective Major Orthopedic Surgery: Quality, Quantity, and Timing.
Improvements to enhanced recovery pathways in orthopedic surgery are reducing the time that patients spend in the hospital, giving an increasingly vital role to prehabilitation and/or rehabilitation after surgery. Nutritional support is an important tenant of perioperative medicine, with the aim to integrate the patient's diet with food components that are needed in greater amounts to support surgical fitness. Regardless of the time available between the time of contemplation of surgery and the day of admission, a patient who eats healthy is reasonably more suitable for surgery than a patient who does not meet the daily requirements for energy and nutrients. Moreover, a successful education for healthy food choices is one possible way to sustain the exercise therapy, improve recovery, and thus contribute to the patient's long-term health. The expected benefits presuppose that the patient follows a healthy diet, but it is unclear which advice is needed to improve dietary choices. We present the principles of healthy eating for patients undergoing major orthopedic surgery to lay the foundations of rational and valuable perioperative nutritional support programs. We discuss the concepts of nutritional use of food, requirements, portion size, dietary target, food variety, time variables of feeding, and the practical indications on what the last meal to be consumed six hours before the induction of anesthesia may be together with what is meant by clear fluids to be consumed until two hours before. Surgery may act as a vital "touch point" for some patients with the health service and is therefore a valuable opportunity for members of the perioperative team to promote optimal lifestyle choices, such as the notion and importance of healthy eating not just for surgery but also for long-term health benefit.


Diamond-shaped optical fiber delivery system.
Physicians who use laser systems are looking for new fiberoptic delivery systems with better performance. We searched for a delivery system with high power and with a tip of high durability that may be reused many times. The delivery systems currently found on the market can burn and break easily. The surgeon may either finish the surgery with a damaged delivery system, or have to switch to a new one during surgery, thus increasing the inconvenience and cost of the surgery. Distal tips of fibers used with Nd:YAG and diode lasers have been shaped and polished according to their specific function, including excision, vaporization, and coagulation of tissue. We have developed several prototypes of optical fiber delivery systems, and in particular, we have developed a prototype of a delivery system, called the Diamond-Shaped Optical Fiber Delivery System, or Diamond. By making the tip of an optical fiber diamond shaped, we are able to obtain greater mechanical resistance than that of other delivery systems that were tested in our laboratory. In the Colorectal Unit, we applied it in surgical procedures such as perianal fistulectomy, hemorrhoidectomy, and excision of condilomata acuminata and pilonidal cyst and sinuses. During cutting and coagulating, the system performed well with no technical problems. A new delivery system (Diamond) gave satisfactory performance during surgical operations. Its main component, the diamond-shaped distal end, provided high durability and was reused many times, thus improving the quality of operations and reducing expenses greatly.


Parents and newborn "togetherness" after birth.
Zero separation is a family-centred approach where newborns should be accompanied by their parents, regardless of the type of birth or health status. To our knowledge, few studies have described the way this approach is realized in clinical practice. This study describes situations of separation between mother/partner and newborn after birth on the labour ward, maternity ward and at the neonatal unit. An observation study was conducted during four months at a Swedish hospital. All caregivers at the three units were given the task of collecting the data. A semantic thematic analysis was performed with an inductive approach. Six themes emerged from the analysis. Two themes were common to all three units, one theme was common to two units and three themes emerged at only one unit. The themes describe various causes of separation, such as organizational and economic barriers, clinical routines, parents' own decisions, shortage of collaboration within and between units, as well as a shortage of interprofessional communication. Our study shows that there is still a gap between the latest evidence-based knowledge of the importance of zero separation and current practice in newborn care. There is a need for continuous collaboration between all units responsible for the care of mother and newborn.


Nitric oxide synthase inhibitors do not substitute in rats trained to discriminate phencyclidine from saline.
Release of nitric oxide occurs as a consequence of glutamate stimulation of NMDA receptors and is dependent upon calcium-calmodulin activation of the enzyme nitric oxide synthase. Since nitric oxide may serve as an intracellular messenger for NMDA glutamatergic neurons, it could be hypothesized that blockade of its synthesis may produce pharmacological effects similar to those of NMDA receptor antagonists. The purpose of the present study was to compare the effects of nitric oxide synthase inhibitors to those of the high affinity NMDA open channel blocker phencyclidine in drug discrimination, a pharmacologically selective procedure in which phencyclidine produces distinctive effects. Rats were trained to discriminate 2 mg/kg phencyclidine from saline in a standard two-lever discrimination task with food reward. Whereas phencyclidine dose-dependently substituted for itself, 7-nitroindazole, L-NAME (N(G)-nitro-L-arginine methyl ester), and L-NOARG (N(G)-nitro-L-arginine) failed to substitute for phencyclidine when administered intraperitoneally. L-NAME and 7-nitroindazole were tested up to doses that disrupted responding, providing evidence that a behaviorally-relevant dosage range was evaluated. Although these results conflict with those of a previous study which found that nitric oxide synthase inhibitors substituted for phencyclidine and produced phencyclidine-like catalepsy in pigeons, they are consistent with research showing that these drugs did not produce phencyclidine-like pharmacological effects in behavioral procedures in rats.


A genome-scale metabolic reconstruction of soybean and Bradyrhizobium diazoefficiens reveals the cost-benefit of nitrogen fixation.
Nitrogen-fixing symbioses allow legumes to thrive in nitrogen-poor soils at the cost of diverting some photoassimilate to their microsymbionts. Effort is being made to bioengineer nitrogen fixation into nonleguminous crops. This requires a quantitative understanding of its energetic costs and the links between metabolic variations and symbiotic efficiency. A whole-plant metabolic model for soybean (Glycine max) with its associated microsymbiont Bradyrhizobium diazoefficiens was developed and applied to predict the cost-benefit of nitrogen fixation with varying soil nitrogen availability. The model predicted a nitrogen-fixation cost of c. 4.13 g C g-1 N, which when implemented into a crop scale model, translated to a grain yield reduction of 27% compared with a non-nodulating plant receiving its nitrogen from the soil. Considering the lower nitrogen content of cereals, the yield cost to a hypothetical N-fixing cereal is predicted to be less than half that of soybean. Soybean growth was predicted to be c. 5% greater when the nodule nitrogen export products were amides versus ureides. This is the first metabolic reconstruction in a tropical crop species that simulates the entire plant and nodule metabolism. Going forward, this model will serve as a tool to investigate carbon use efficiency and key mechanisms within N-fixing symbiosis in a tropical species forming determinate nodules.


A phase II trial of piroxantrone in endometrial cancer: Southwest Oncology Group study 8918.
A phase II trial of the new anthrapyrazole piroxantrone was carried out by the Southwest Oncology Group in patients with advanced metastatic or recurrent endometrial cancer. A two-stage statistical design targeted accrual of 20 eligible patients. The starting dose of piroxantrone was 150 mg/m2 in patients without prior radiation therapy (RT) and 120 mg/m2 in patients with prior RT. There were 15 eligible patients, six of whom had received prior hormonal therapy while nine patients had not received prior hormonal therapy. Eight patients had received prior RT while seven patients had not received any prior RT. One to seven cycles of piroxantrone were administered. Dose escalation was feasible in four patients. No grade 5 toxicity was experienced by any patients. Most of the grade 4 (granulocytopenia in one) and grade 3 (leukopenia in three, granulocytopenia in three, anemia in two and thrombocytopenia in one) toxicity was related to myelosuppression. Grade 3 non-hematologic toxicities were nausea, fatigue and SGOT elevation. There was one partial response for a response rate of 7% (95% CI 0.2-32%) and median survival was 11 months (95% CI 3-13 months). The study was prematurely terminated due to lack of patient accrual.


Host-parasite interaction in the rat renal pelvis: a possible role for pili in the pathogenesis of pyelonephritis.
The initial interaction between bacteria and the renal pelvic epithelium may determine whether intrarenal infection occurs. A model of retrograde pyelonephritis was employed to study these events by electron microscopy. Female rats received an intravesicular inoculation of a 0.5-ml suspension of Proteus mirabilis containing 10(8) organisms. At intervals after inoculation, the kidneys were fixed by intravascular perfusion and the tissues were prepared for electron microscopy. During the first 24 h, increasing numbers of bacteria were seen to be attached by pili to the renal pelvic epithelial cells. The organism appeared to cross the mucosal barrier by several mechanisms: (a) penetration into the cytoplasm of intact epithelial cells, (b) passage between epithelial cells that were separated by excessive hydrostatic pressure generated during bladder inoculation, (c) passage across necrotic regions of the pelvis, and (d) translocation to the cortex by calicotubular backflow. Whereas at inoculation bacteria possessed pili 40 A in diameter (type III pili) 24 h after reflux, the predominant type of pili measured 70 A in thickness (type IV pili). Repetitive subculture induced a similar transition in vitro. To assess the influence of pili type on virulence in this model, 80 rats were challenged with either type III or type IV pilated organisms and the frequency of rats with cortical abscesses were compared at 1 wk. A significantly greater number of rats inoculated with type IV pilated Proteus manifested macroscopic evidence of infection. These results suggest that pili play a role in the pathogenesis of ascending pyelonephritis.


Epidemiology of Breast Cancer Mortality in Kazakhstan, trends and Geographic Distribution.
According to the International Agency for Research on Cancer, ongoing demographic changes will lead to an increase in the number of deaths from breast cancer (BC) per year in the vast majority of regions. In 2040 it is expected that 1.04 million people worldwide will die from this malignancy, including 2,380 women in Kazakhstan. The retrospective study (2009-2018) was done using descriptive and analytical methods of oncoepidemiology. The extensive, crude and age-specific incidence rates are determined according to the generally accepted methodology used in sanitary statistics. The data were used to calculate the average percentage change (APС) using the Joinpoint regression analysis to determine the trend over the study period. During 10 years 12,958 women died from BC. An average age of the death was 61.6 years (95%CI=60.6-62.6) and tended to increase (APC=+0.6%, R2=0.6117). Age-specific rates had a bimodal increase with peak rates at 70-74 years - 76.7±5.5 (APC=+3.4%, R2=0.2656) and 80-84 years - 78.0±9.1 (APC=+3.7%, R2=0.0875). The age-standardized rate was 13.9 per 100,000 of female population, and the trend has decreased. When compiling thematic maps, mortality rates were determined on the basis of standardized indicators: low - up to 12.5, average - from 12.5 to 15.2, high - above 15.2 per 100,000. The results of the spatial analysis showed the regions with a higher levels of BC mortality rate per 100,000: Pavlodar (16.9), Almaty (19.2) and Astana cities (19.3). Age-standardized mortality rates had a strong downward trend (APC=-4.0%, R2=0.9218). The decrease mostly is due to a large coverage of the population by mammography screening and to an improvement in the effectiveness of breast cancer treatment.


Nerve regenerative effects of GABA-B ligands in a model of neuropathic pain.
Neuropathic pain arises as a direct consequence of a lesion or disease affecting the peripheral somatosensory system. It may be associated with allodynia and increased pain sensitivity. Few studies correlated neuropathic pain with nerve morphology and myelin proteins expression. Our aim was to test if neuropathic pain is related to nerve degeneration, speculating whether the modulation of peripheral GABA-B receptors may promote nerve regeneration and decrease neuropathic pain. We used the partial sciatic ligation- (PSL-) induced neuropathic model. The biochemical, morphological, and behavioural outcomes of sciatic nerve were analysed following GABA-B ligands treatments. Simultaneous 7-days coadministration of baclofen (10 mg/kg) and CGP56433 (3 mg/kg) alters tactile hypersensitivity. Concomitantly, specific changes of peripheral nerve morphology, nerve structure, and myelin proteins (P0 and PMP22) expression were observed. Nerve macrophage recruitment decreased and step coordination was improved. The PSL-induced changes in nociception correlate with altered nerve morphology and myelin protein expression. Peripheral synergic effects, via GABA-B receptor activation, promote nerve regeneration and likely ameliorate neuropathic pain.


Cytokine and cytokine-like inflammation markers, endothelial dysfunction, and imbalanced coagulation in development of diabetes and its complications.
Recent developments indicate that pathophysiological mechanisms leading to beta-cell damage, insulin resistance, and the vascular complications of diabetes include an activation of the inflammation cascade, endothelial dysfunction, and procoagulant imbalance. Their circulating biomarkers may therefore provide opportunities for early diagnosis and targets for novel treatments. Circulating biomarkers of these pathways such as TNFalpha, IL-6, C-reactive protein (CRP) (inflammation), vascular cellular adhesion molecule-1, interstitial cellular adhesion molecule-1, E-selectin, von Willebrand factor (endothelial dysfunction), plasminogen activator inhibitor-1, fibrinogen, P-selectin (procoagulant state), and adiponectin (antiinflammation) may be associated with development of both type 1 and type 2 diabetes and some studies, particularly in type 2 diabetes, have demonstrated that certain biomarkers may have independent predictive value. Similarly studies have shown that these biomarkers may be associated with development of diabetic nephropathy and retinopathy, and again, particularly in type 2 diabetes, with cardiovascular events as well. Finally, the comorbidities of diabetes, namely obesity, insulin resistance, hyperglycemia, hypertension and dyslipidemia collectively aggravate these processes while antihyperglycemic interventions tend to ameliorate them. Increased CRP, IL-6, and TNFalpha, and especially interstitial cellular adhesion molecule-1, vascular cellular adhesion molecule-1, and E-selectin are associated with nephropathy, retinopathy, and cardiovascular disease in both type 1 and type 2 diabetes. Whereas further work is needed, it seems clear that these biomarkers are predictors of increasing morbidity in prediabetic and diabetic subjects and should be the focus of work testing their clinical utility to identify high-risk individuals as well as perhaps to target interventions.


Giant Thermal Rectification from Polyethylene Nanofiber Thermal Diodes.
The realization of phononic computing is held hostage by the lack of high-performance thermal devices. Here, it is shown through theoretical analysis and molecular dynamics simulations that unprecedented thermal rectification factors (as large as 1.20) can be achieved utilizing the phase-dependent thermal conductivity of polyethylene nanofibers. More importantly, such high thermal rectifications only need very small temperature differences (<20 °C) across the device, which is a significant advantage over other thermal diodes which need temperature biases on the order of the operating temperature. Taking this into consideration, it is shown that the dimensionless temperature-scaled rectification factors of the polymer nanofiber diodes range from 12 to 25-much larger than those of other thermal diodes (<8). The polymer nanofiber thermal diode consists of a crystalline portion whose thermal conductivity is highly phase-sensitive and a cross-linked portion which has a stable phase. Nanoscale size effect can be utilized to tune the phase transition temperature of the crystalline portion, enabling thermal diodes capable of operating at different temperatures. This work will be instrumental to the design of high performance, inexpensive, and easily processible thermal devices, based on which thermal circuits can be built to ultimately enable phononic computing.


Mate choice for genetic quality when environments vary: suggestions for empirical progress.
Mate choice for good-genes remains one of the most controversial evolutionary processes ever proposed. This is partly because strong directional choice should theoretically deplete the genetic variation that explains the evolution of this type of female mating preference (the so-called lek paradox). Moreover, good-genes benefits are generally assumed to be too small to outweigh opposing direct selection on females. Here, we review recent progress in the study of mate choice for genetic quality, focussing particularly on the potential for genotype by environment interactions (GEIs) to rescue additive genetic variation for quality, and thereby resolve the lek paradox. We raise five questions that we think will stimulate empirical progress in this field, and suggest directions for research in each area: (1) How is condition-dependence affected by environmental variation? (2) How important are GEIs for maintaining additive genetic variance in condition? (3) How much do GEIs reduce the signalling value of male condition? (4) How does GEI affect the multivariate version of the lek paradox? (5) Have mating biases for high-condition males evolved because of indirect benefits?


Exceeding the resolving imaging power using environmental conditions.
We present two approaches that use the environmental conditions in order to exceed the classical Abbe's limit of resolution of an aperture-limited imaging system. At first we use water drops in order to improve the resolving capabilities of an imaging system using a time-multiplexing approach. The limit for the resolution improvement capabilities is equal to the size of the rain drops. The rain drops falling close to the imaged object act as a sparse and random high-resolution mask attached to it. By applying proper image processing, the center of each falling drop is located, and the parameters of the encoding grating are extracted from the captured set of images. The decoding is done digitally by applying the same mask and time averaging. In many cases urban environment includes periodic or other high-resolution objects such as fences. Actually urban environment includes many objects of this type since from an engineering point of view they are considered appealing. Those objects follow well known standards, and therefore their structure can be a priori known even without being fully capable of imaging them. We show experimentally how we use such objects in order to superresolve the contour of moving targets passing in front of them.


B-A Chromosome Translocations Possessing an A Centromere Partly Overcome the Root-Restricted Process of Chromosome Elimination in Aegilops speltoides.
Some eukaryotes exhibit dramatic genome size differences between cells of different organs, resulting from the programmed elimination of chromosomes. Aegilops speltoides is an annual diploid species from the Poaceae family, with a maximum number of eight B chromosomes (Bs) in addition to its inherent seven pairs of standard A chromosomes (As). The Bs of this species undergo precise elimination in roots early in embryo development. In areal parts of the plant, the number of Bs is stable. To affect the root restricted process of B chromosome elimination, we employed X-ray mutagenesis, and different types of restructured Bs were identified. Standard Bs were observed in all analyzed shoots of mutagenized plants, while B-A translocations were only observed in 35.7% of F1 plants. In total 40 different B variants inconsistently escaped the elimination process in roots. As a result, mosaicism of B chromosome variants was found in roots. Only a small B chromosome fragment fused to an A chromosome was stably maintained in roots and shoots across F1 to F3 generations. The absence of B-A translocation chromosomes possessing a derived B centromere in root cells implies that the centromere of the B is a key component of the chromosome elimination process.


Access to infertility care in Brazil: validation of a questionnaire for a bioethical discussion.
The World Health Organization characterizes infertility as a disease since 2009; however, in many countries, reproductive health is not prioritized. Characterizing a target population and knowing the barriers to accessing reproductive care may allow for a broadening of the discussion on how to provide equal opportunities. The objective of this study was to develop and validate a questionnaire that seeks to identify socioeconomic and cultural characteristics of Brazilian infertile couples to open the discussion on the principle of fair access to health. (1) literature review in the main databases, (2) questionnaire elaboration by researchers within the areas of human reproduction and bioethics, concerned with content adequacy and comprehension, using the Google Forms online platform, (3) pilot study - the questionnaire was applied to 54 couples, over 18 years of age, that were seeking treatment for infertility and accepted to participate in the study and (4) statistical analysis - for continuous numerical variables, mean, standard deviation and 95% CI of the means were presented. For discrete numerical variables, median, interquartile range, minimum and maximum were presented. Forty-four questionnaires were fully completed and with adequate answers. The questionnaire proved to be objective and easy to understand. It was possible to obtain information on education, race of the couple, the impact of infertility on the couple's life, socioeconomic conditions, and the main difficulties in accessing medical care for treatment of infertility. The questionnaire proved to be feasible in collecting appropriate information to characterize a target population and the only limitation was that there was no academic expert evaluation prior to the pilot test.


The ultrastructure of feline infectious peritonitis virus in feline embryonic lung cells.
The ultrastructure of feline infectious peritonitis virus in cultured feline embryonic lung cells is reported. Feline embryonic lung cells were infected with feline infectious peritonitis virus and studied by transmission electron microscopy. The virus was not apparent in the cultured cells until 24 h after infection when it occurred in the endoplasmic reticulum, perinuclear space, Golgi apparatus, free in the cytoplasm, in large vacuoles in the cytoplasm and outside the cell membrane. The virus possessed typical coronavirus morphology and was produced by budding into the endoplasmic reticulum. There was no evidence to indicate that this virus budded through the cell membrane. Multinucleate giant cells were formed by infection of the cultured cells with the virus. The host cells were destroyed by the virus and phagocytosed by apparently healthy cells.


[Long-term consequences of esophageal atresia; esophageal and lung abnormalities in adulthood].
Esophageal atresia is a rare congenital anomaly. Due to increased survival rates, the population of adults born with this malformation is growing. These patients turn out to have an increased risk to develop Barrett's esophagus, esophageal carcinoma or lung abnormalities like bronchiectasis. This is illustrated by three cases: a 42-year-old man with an irresectable esophageal squamous cell carcinoma; a 23-year-old man with a Barrett's esophagus without any reflux complaints; and a 51-year-old women with a reflux esophagitis and extensive bronchiectasis due to a combination of gastroesophageal reflux with chronic aspiration and a reduced sputum clearance because of a history of tracheomalacia. It is important for healthcare providers to be aware of these risks and the possible absence of symptoms, in order to detect abnormalities at an early stage and improve quality of life of these patients.


[Intraocular malignant lymphoma].
The paper in devoted to a rare condition difficult to diagnose, intraocular lymphoma. Five patients with a disease history of 2 to 6 years were examined. The authors describe clinical features of the condition and histologic and electron microscopic findings. The clinical pattern of malignant intraocular lymphoma in polymorphous, therefore virtually every case with a sluggish uveitis or posterior scleritis resistant to common therapy has to be verified. A hematologist should take part in examination of such patients. Since the involvement is local, as our findings evidence, a course of polychemotherapy is necessary for such patients.


Safety of foods derived from genetically modified plants.
Biopharmaceuticals have been available for clinical use for nearly three decades, but foods derived from agribiotechnology have been available for just under a decade. Controversy surrounding foods from genetically modified (GM) plants has focused primarily upon their allergenicity, with lesser concerns about antibiotic resistance genes. Concerns are related to possible environmental impacts on non-human species, including effects on non-target species (e.g., butterflies) and on the development of so-called "super weeds." Food allergies are no more prevalent in foods from GM plants than in conventional foods. Further, the use of antibiotics in the development of GM plants does not pose a significant risk to the human population. Foods from the current GM plant products have been shown not to pose any detrimental effects to humans, and, in fact, nutritionally enhanced products are being developed. GM foods are subjected globally to intense regulatory scrutiny, and extensive data have been provided consistently to regulatory agencies in the United States on a voluntary basis, with mandatory reporting of data soon to be in force. Existing environmental concerns appear to be unjustified on the basis of existing data and experience.


The emergence of sarcopenia as an important entity in older people.
Sarcopenia refers to the loss of muscle mass and strength seen with advancing age. The pathophysiology is multifactorial, with loss of muscle satellite cells, changes in hormonal systems, chronic inflammation, oxidative stress and anabolic resistance to protein utilisation all implicated. Older age, female sex and immobility are important risk factors. Sarcopenia is clinically important as it is a major risk factor for physical frailty, falls, prolonged hospitalisation, dependency and earlier death. Diagnosis requires evidence of reduced muscle mass measured by handgrip strength or walk speed, together with evidence of low muscle mass, measured by one of a variety of techniques such as bioimpedance analysis or dual X-ray absorptiometry. Resistance training is the only intervention of proven efficacy to treat sarcopenia, but a range of nutritional and pharmacological interventions are under test, including myostatin inhibitors, leucine and protein supplementation, angiotensin-converting enzyme inhibitors and allopurinol.


The maladroit adolescent: learning disorders and attentional deficits.
Being maladroit in a society with increasing emphasis on performance is a formidable challenge to adolescent development. Helping maladroit adolescents achieve optimal potential is a challenge for physicians, educators, and others involved in their care. The first steps for professionals are recognizing SLD and ADD and understanding the configurations of these problems as they apply to adolescents and adults. Educators can prioritize academic concerns and evaluate and provide intervention strategies. Physicians can review new pharmacologic data and follow-up studies to help formulate clinical judgments, particularly around the area of medication. Along with counselors, physicians will provide better longitudinal care when aware of the social and behavioral outcomes reported for patients similar to theirs. Finally, awareness of resources facilitates multidisciplinary communication and provides access for patients and clients to needed medical, educational, and support resources. There has always been a sense of advocacy among those working with the adolescent whose exaggerated maladroitness stems from a problem with learning or attention. Perhaps this review will provide a data base from which advocacy can expand to specific action. That has been our goal.


Getting REDD-y: conservation and climate change in Latin America.
Deforestation in Latin America, especially in the Amazon basin, is a major source of greenhouse gases such as carbon dioxide that contribute to global warming. Protected areas play a vital role in minimizing forest loss and in supplying key environmental services, including carbon sequestration and rainfall regulation, which mitigate the adverse impacts of climate change amid a rising tide of economic development in the region. The area of protected forest has expanded rapidly since 1980 to cover one-fifth of Latin America and more than two-fifths of Amazonia, a region whose rain forest captures some 40 percent of Latin America's carbon emissions. The reserve sector has traditionally suffered from severe underfunding, but the possibility of new resources being generated through financial compensation for "reduced emissions from deforestation and forest degradation" (REDD) or "avoided deforestation" under a new Kyoto protocol after 2012 could help strengthen the environmental and social roles of protected areas. However, a number of major implementation and governance challenges will need to be addressed.


A 22-year-old woman with chronic, mild, intermittent knee pain.
The following case is presented to illustrate the roentgenographic and clinical findings of a condition of interest to orthopedic surgeons. The initial history, physical findings, and roentgenographic examination are found on the first page. The final clinical and roentgenographic diagnosis is presented on the following page.


Bimanual Coordination Learning with Different Augmented Feedback Modalities and Information Types.
Previous studies have shown that bimanual coordination learning is more resistant to the removal of augmented feedback when acquired with auditory than with visual channel. However, it is unclear whether this differential "guidance effect" between feedback modalities is due to enhanced sensorimotor integration via the non-dominant auditory channel or strengthened linkage to kinesthetic information under rhythmic input. The current study aimed to examine how modalities (visual vs. auditory) and information types (continuous visuospatial vs. discrete rhythmic) of concurrent augmented feedback influence bimanual coordination learning. Participants either learned a 90°-out-of-phase pattern for three consecutive days with Lissajous feedback indicating the integrated position of both arms, or with visual or auditory rhythmic feedback reflecting the relative timing of the movement. The results showed diverse performance change after practice when the feedback was removed between Lissajous and the other two rhythmic groups, indicating that the guidance effect may be modulated by the type of information provided during practice. Moreover, significant performance improvement in the dual-task condition where the irregular rhythm counting task was applied as a secondary task also suggested that lower involvement of conscious control may result in better performance in bimanual coordination.


Dihydrodehydrodiisoeugenol enhances adipocyte differentiation and decreases lipolysis in murine and human cells.
Loss of subcutaneous fat is a hallmark of ageing usually starting in the face. Attempts to ameliorate cosmetically the appearance of subcutaneous fat loss have been of limited success as they fail to rebuild the missing subcutaneous tissue. Ageing-driven loss of subcutaneous fat results from (i) the reduced capacity of pre-adipocytes to differentiate into adipocytes and (ii) the fact that adipocytes of the elderly secrete increased amounts of TNFα, that in turn enhances lipolysis, inhibits pre-adipocyte differentiation and induces dedifferentiation of adipocytes. The neolignan dihydrodehydrodiisoeugenol (DDE) caused a 30% increase in lipid accumulation in murine 3T3-L1 cells. This effect was accompanied by an induction of the differentiation-associated transcription factors peroxisome proliferator-activated receptorγ (PPARγ2), CAAT/enhancer-binding protein α (C/EBPα), fatty acid binding protein 4 and adiponectin, and a loss of the pre-adipocyte marker Pref1. In addition, DDE diminished both basal and TNFα-induced lipolysis. Similar results were obtained in human subcutaneous (hsc) pre-adipocytes cultured in an age-adapted hormone mix with reduced levels of insulin and dexamethasone. In this system, DDE significantly increased lipid accumulation by 71% and 94% and was associated with an induction of PPARγ2 and adiponectin mRNA expression. DDE also reduced basal lipolysis in mature hsc adipocytes. DDE acted as a partial PPARγ agonist because (i) DDE displaced PPARγ ligand from the human PPAR ligand-binding site, (ii) DDE-induced lipid accumulation and (iii) DDE-induced adiponectin secretion could be overcome by the addition of PPARγ antagonists. Taken together, these studies identify DDE as a compound well suited to prevent and reverse loss of subcutaneous fat.


[Rosacea - manifestations and treatment options].
Rosacea - manifestations and treatment options Abstract. Rosacea is a common dermatosis of the face with a prevalence of up to 22 %, according to the current literature. The known trigger factors include caffeine, alcohol, sunlight, hot and spicy foods, psychological stress, menstruation and extreme temperatures or temperature fluctuations. Diagnosis is most often clinical, however, due to the numerous differential diagnoses, performing a biopsy may be helpful in atypical manifestations. Depending on the symptoms, in addition to the avoidance of trigger factors, physical therapeutic options as well as topical and systemic drugs are available.


Experience of parents with head and neck cancer who are caring for young children.
This paper is a report of a study exploring the experience and support needs of parents with head and neck cancer, who are also caring for their young children. The incidence of head and neck cancer is steadily rising in the younger age groups (under 50 years). As a direct consequence, more parents of young children are being diagnosed with this cancer. Using a qualitative descriptive design, 12 patients with young children under the age of 16 were interviewed during 2008. The data were analysed using cognitive mapping. At diagnosis, parents experienced the fear of missing milestones in the life of their children and fear of telling the children about their diagnosis. During treatment, parents' inevitable stays in hospital led to separation from the children and the need to prepare their children for the first hospital visit. Parents experienced many debilitating effects of treatment, which had an impact on family activities, especially those that involved eating. There was often changing roles within the family. After treatment, the experience of living with cancer for parents resulted in the desire to get back to day-to-day life. Parents had an increased appreciation of life but were also living with uncertainties. The most important support networks for parents were their spouse, family and friends. Oncology nurses are in an excellent position to offer support and guidance to parents with cancer who have children, and should encourage them to explore with the family the best way of managing role change during treatment.


Machine Learning Modeling Based on Microbial Community for Prediction of Natural Attenuation in Groundwater.
Natural attenuation is widely adopted as a remediation strategy, and the attenuation potential is crucial to evaluate whether remediation goals can be achieved within the specified time. In this work, long-term monitoring of indigenous microbial communities as well as benzene, toluene, ethylbenzene, and xylene (BTEX) and chlorinated aliphatic hydrocarbons (CAHs) in groundwater was conducted at a historic pesticide manufacturing site. A machine learning approach for natural attenuation prediction was developed with random forest classification (RFC) followed by either random forest regression (RFR) or artificial neural networks (ANNs), utilizing microbiological information and contaminant attenuation rates for model training and cross-validation. Results showed that the RFC could accurately predict the feasibility of natural attenuation for both BTEX and CAHs, and it could successfully identify the key genera. The RFR model was sufficient for the BTEX natural attenuation rate prediction but unreliable for CAHs. The ANN model showed better performance in the prediction of the attenuation rates for both BTEX and CAHs. Based on the assessments, a composite modeling method of RFC and ANN was proposed, which could reduce the mean absolute percentage errors. This study reveals that the combined machine learning approach under the synergistic use of field microbial data has promising potential for predicting natural attenuation.


[Methods of hemofiltration].
The methods of continuous hemofiltration therapy are described. The various forms of blood access are discussed, especially the pumpdriven venovenous form of hemofiltration utilizing a double lumen catheter in the superior caval vein is demonstrated as an alternative to the spontaneous arteriovenous method. The own experiences in over 140 patients with renal, cardial and pulmonal indications are summarized.


Physiologically-based toxicokinetic modeling of durene (1,2,3,5-tetramethylbenzene) and isodurene (1,2,4,5-tetramethylbenzene) in humans.
Physiologically-based toxicokinetic (PB-TK) models are developed to simulate absorption, distribution and excretion of xenobiotics. PB-TK models consist of several groups of compartments, where tissues are grouped together according to physiological parameters (tissue blood flows, tissue group volumes) and physicochemical properties (partition coefficients, metabolic constants). Tetramethylbenzene (TETMB), a mixture of its three isomers: prenitene (1,2,3,4-TETMB), isodurene (1,2,3,5-TETMB), and durene (1,2,4,5-TETMB) is an essential component of numerous commercial preparations of organic solvents. The aim of the study was to develop the PB-TK model for two TETMB isomers, durene and isodurene, in humans. The assumed PB-TK model groups organs and tissues into five physiological compartments: fat tissue, muscles, organs, liver, and brain. The brain has been considered as a separate compartment due to the potential neurotoxicity of TETMB. Water/air, oil/air and blood/air partition coefficients for durene and isodurene were measured in vitro. Tissue/air partition coefficients were calculated from values of olive/air and water/air partition coefficients and the average fat and water content in different tissues. Tissue/blood partition coefficients were calculated as a tissue/air quotient and the blood/air partition coefficient measured in vitro. The Michaelis-Menten constant (KM) values and maximum metabolism rate constant (VMAX) for selected metabolites of durene and isodurene were obtained in vitro using microsomal fraction of the human liver. The developed model was validated against experimental data obtained earlier as a result of an 8-h exposure of volunteers to durene and isodurene vapors of 10 and 25 mg/m3. The prediction of both TETMB isomers concentration in blood as well as of the elimination rates of 2,4,5-TMBA and 2,3,5-TMBA were close to the results of experimental exposures. Simulations of one working week inhalation exposure to aromatic hydrocarbons indicate that the elaborated PB-TK model may be used to predict the chemical distribution in different body compartments, based on physicochemical properties.


Common variants in the periostin gene influence development of atherosclerosis in young persons.
We investigated the influence of genetic variants (rare and common) in the gene encoding periostin (POSTN) on atherosclerosis as measured in arterial specimens from the Pathobiological Determinants of Atherosclerosis in Youth (PDAY) study. A comprehensive survey of common POSTN variants (87 single-nucleotide polymorphisms [SNPs]) in PDAY subjects (n = 2527) identified numerous SNPs associated with raised lesions in abdominal aorta and with fatty streaks in thoracic aorta. These SNPs belonged to a small number of correlation bins that spanned the entire locus. To examine effects of rare variants, we resequenced POSTN functional regions in PDAY cases with raised lesions (n = 291) and controls with no raised lesions (n = 294). However, we found no significant associations with case-control status for carriers of POSTN rare variants using the weighted-sum method for rare variant analysis. We identified common variants in POSTN that are associated with arterial lesions in young persons from the PDAY study. This finding strongly supports a role for periostin in atherogenesis, as suggested by recent proteomics analysis that found abundant expression of periostin in atherosclerotic lesions. Genetic variation may influence atherosclerosis via periostin's known involvement in multiple relevant pathways, including angiogenesis, vascular remodeling, and stimulation of migration and differentiation of vascular smooth muscle cells.


Mechanisms of the stimulation of insulin release by oxytocin in normal mouse islets.
Oxytocin (OT) produced a dose-dependent increase in somatostatin, glucagon and insulin release by isolated mouse islets. A small effect on somatostatin release was observed with 0.1 nM-OT, but 1-10 nM-OT was required to affect A- and B- cells significantly. The effects of OT on somatostatin and glucagon release were similar in the presence of 3 mM- and 10 mM-glucose. No change in insulin release was produced by OT in 3 mM-glucose, but a stimulation was still observed in the presence of a maximally effective concentration of glucose (30 mM). The increase in insulin release produced by OT (in 15 mM-glucose) was accompanied by small accelerations of 86Rb and 45Ca efflux from islet cells. Omission of extracellular Ca2+ accentuated the effect of OT on 86Rb efflux, attenuated that on 45Ca efflux, and abolished that on release. OT never inhibited 86Rb efflux. It did not affect the resting potential of B-cells, but slightly increased the Ca2(+)-dependent electrical activity induced by 15 mM-glucose. OT did not affect cyclic AMP levels, but increased inositol phosphate levels in islet cells. It is suggested that the amplification of glucose-induced insulin release that OT produces is due to a stimulation of phosphoinositide metabolism, and presumably an activation of protein kinase C, rather than to a change in cyclic AMP levels or a direct action on the membrane potential. Since OT is present in the pancreas, it is possible that it exerts a neuropeptidergic control of the islet function.


Magnetic resonance spectroscopy of live Drosophila melanogaster using magic angle spinning.
High-Resolution Magic Angle Spinning (HRMAS) proton magnetic resonance spectroscopy ((1)H-MRS) is a novel non-destructive technique that improves spectral line-widths and allows high-resolution spectra to be obtained from extracts, intact cells, cell cultures, and more importantly intact tissue to investigate relationships between metabolites and cellular processes. In vivo HRMAS (1)H-MRS studies have yet to be reported in the live fruit fly Drosophila melanogaster. Drosophila, as a simpler genetic organism, allows the multiple biological functions and various evolutionarily conserved signaling pathways to be examined at the whole organism level and it is a useful model for investigating genetics and physiology. To this end, we developed and implemented an in vivo HRMAS (1)H-MRS method to investigate live Drosophila at 14.1 T. Here, we outline an HRMAS (1)H-MRS protocol for the molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe. This technique is a novel, in vivo, non-destructive Drosophila metabolite measurement approach, which enables the identification of disease biomarkers and thus may contribute to novel therapeutic development.


Integrated recovery management model for ex-offenders with co-occurring mental health and substance use disorders and high rates of HIV risk behaviors.
This paper provides outcomes from an evaluation of a federally funded program combining HIV prevention services with an integrated mental health and substance abuse treatment program to a population of primarily African American ex-offenders living with, or at high risk for contracting HIV in Memphis, Tennessee. During the 5-year evaluation, data were collected from 426 individuals during baseline and 6-month follow-up interviews. A subset of participants (n = 341) completed both interviews. Results suggest that the program was successful in reducing substance use and mental health symptoms but had mixed effects on HIV risk behaviors. These findings are important for refining efforts to use an integrated services approach to decrease (a) the effects of substance use and mental health disorders, (b) the disproportionate impact of criminal justice system involvement, and (c) the HIV infection rate in African American ex-offenders in treatment.


Interfacial bonding in a CdS/PVA nanocomposite: A Raman scattering study.
Raman spectroscopy is employed to characterize the bonding between CdS nanoparticles (NPs) and a polyvinyl alcohol (PVA) as well as structural changes in the polymeric matrix caused by incorporation of NPs. It is shown that after the formation of CdS NPs the vibrations of carbonyl groups in acetate residuals of PVA and of C-O groups at the macromolecules ends disappear. Formation of NPs also leads to an increased degree of hydrogen bonding and crystallinity of the hybrid material as compared with the unloaded polymer. The observed changes are ascribed to the formation of coordinative bonds and hydrogen between the CdS nanoparticles and polymeric macromolecules. The scheme of this interfacial bonding is also proposed.


Researcher perspectives on including people who use drugs in clinical research.
Background: People who use drugs are commonly excluded from clinical research despite being disproportionately impacted by numerous health conditions. Recent work indicates that community-recruited individuals report low support of research inclusion for persons reporting substance use.Objective: This study conducted a mixed-method assessment of researchers' attitudes on including persons reporting substance use in clinical research.Methods: Respondents (N = 168; 58% female; 58% psychologists) recruited via scientific society listservs and Twitter completed a survey containing two hypothetical research vignettes. Quantitative items evaluated respondents' endorsement of research participation by healthy adults, people who use drugs, and vulnerable populations. Qualitative items included open-ended questions asking reasons why people who use drugs should and should not participate in research.Results: Respondents reported significantly lower support for research participation by people who use drugs compared to healthy adults (p <.001). Open-ended responses concerning the inclusion of people who use drugs included themes relevant to the Belmont Principles (e.g., capacity to consent) and data quality (e.g., "bad data," poor compliance).Conclusion: Although lower support for research participation by people who use drugs was observed compared to healthy adults, the magnitude of this difference was smaller than reported for prior community-recruited respondents. These findings emphasize salient factors that may serve as both protections for and barriers to inclusion of people who use drugs in research. Initiatives including adoption of person-first language, addressing stereotyping of people who use drugs, and emphasizing the benefits of including these populations in clinical research should be explored to reduce bias while retaining needed protections.


The effect of MMF dose and trough levels on adverse effects in pediatric heart transplant recipients.
Limited pharmacokinetic and safety data exist for MMF in pediatric HTR. Previously targeted MPA-TL are 1.5-3.0 μg/mL. The objective of this study was to assess the outcomes targeting MPA-TL of 0.8-2.0 μg/mL in pediatric HTR. MPA-TL were retrospectively collected 2-12 months post-transplant. Acute rejection, infection, leukopenia, and GI complaints were then correlated with MPA-TL. A total of 355 MPA-TL from 22 HTR were included. Median age was 2.5 yr. Primary indication for transplant was dilated cardiomyopathy (64%). Mean MPA-TL was 1.7 ± 0.9 μg/mL. African American patients received significantly higher doses (702 ± 235 mg/m(2) ) compared with other races (p = 0.035). Leukopenia was less common in patients with SUB MPA vs. others (p = 0.01). MMF was discontinued for GI complaints in one patient and leukopenia in two patients. One SUB patient had acute rejection, and one SUP patient had infection. One-yr survival was 100%. Targeting a lower range for MPA-TL was not associated with significant rejection or infection. Despite lower MPA-TL, MMF was discontinued in 3/22 patients for adverse effects.


Dual-core steered non-rigid registration for multi-modal images via bi-directional image synthesis.
In prostate cancer radiotherapy, computed tomography (CT) is widely used for dose planning purposes. However, because CT has low soft tissue contrast, it makes manual contouring difficult for major pelvic organs. In contrast, magnetic resonance imaging (MRI) provides high soft tissue contrast, which makes it ideal for accurate manual contouring. Therefore, the contouring accuracy on CT can be significantly improved if the contours in MRI can be mapped to CT domain by registering MRI with CT of the same subject, which would eventually lead to high treatment efficacy. In this paper, we propose a bi-directional image synthesis based approach for MRI-to-CT pelvic image registration. First, we use patch-wise random forest with auto-context model to learn the appearance mapping from CT to MRI domain, and then vice versa. Consequently, we can synthesize a pseudo-MRI whose anatomical structures are exactly same with CT but with MRI-like appearance, and a pseudo-CT as well. Then, our MRI-to-CT registration can be steered in a dual manner, by simultaneously estimating two deformation pathways: 1) one from the pseudo-CT to the actual CT and 2) another from actual MRI to the pseudo-MRI. Next, a dual-core deformation fusion framework is developed to iteratively and effectively combine these two registration pathways by using complementary information from both modalities. Experiments on a dataset with real pelvic CT and MRI have shown improved registration performance of the proposed method by comparing it to the conventional registration methods, thus indicating its high potential of translation to the routine radiation therapy.


Craniopharyngioma in a boy with centronuclear (myotubular) myopathy: clinical and postmortem findings.
This report describes a case of centronuclear myopathy which was considered to represent an example of severe x-linked myotubular myopathy [van Wijngaarden et al. 1969]. The neonatal period was dramatic with weak movements and respiratory problems. This was followed by a gradual improvement of muscle strength but he continued to have severe respiratory problems. At the age of four he developed the symptoms of a craniopharyngioma, which was operated upon. He died from respiratory failure two months after the operation.


Expression of metallothionein gene at different time in testicular interstitial cells and liver of rats treated with cadmium.
Rodent testes are generally more susceptible to cadmium (Cd)-induced toxicity than liver. To clarify the molecular mechanism of Cd-induced toxicity in testes, we compared metallothionein (MT) gene expression, MT protein accumulation, and Cd retention at different time in freshly isolated testicular interstitial cells and liver of rats treated with Cd. Adult male Sprague-Dawley rats weighing 250-280 g received a s.c injection of 4.0 micromol Cd/kg and were euthanized by CO(2) asphyxiation 1 h, 3 h, 6 h, or 24 h later. Tissue was sampled and testicular interstitial cells were isolated. There were three replicates per treatment and 3 animals per replicate for RNA analyses, others, three replicates per treatment and one animal per replicate. MT1 and MT2 mRNA levels were determined by semi-quantitative RT-PCR analysis followed by densitometry scanning, and MT was estimated by the enzyme-linked immunosorbent assay (ELISA) method. Cadmium content was determined by atomic absorption spectrophotometry. The same parametersd were also analyzed in the liver, since this tissue unquestionably accumulate MT. The rat testis expressed MT1 and MT2, the major isoforms. We also found that untreated animals contained relatively high basal levels of both isoform mRNA, which were increased after Cd treatment in liver and peaked at 3 h, followed by a decline. In contrast, the mRNA levels in interstitial cells peaked at 6 h. Interestingly, the induction of MT1 mRNA was lower than MT2 mRNA in liver of rat treated with Cd, but it was opposite to interstitial cells. Cd exposure substantially increased hepatic MT (3.9-fold increase), but did not increase MT translation in interstitial cells. Cd-induced expression of MT isoforms is not only tissue dependent but also time-dependent. The inability to induce the metal-detoxicating MT-protein in response to Cd, may account for a higher susceptibility of testes to Cd toxicity and carcinogenesis compared to liver.


Rapid Thermostabilization of Bacillus thuringiensis Serovar Konkukian 97-27 Dehydroshikimate Dehydratase through a Structure-Based Enzyme Design and Whole Cell Activity Assay.
Thermostabilization of an enzyme with complete retention of catalytic efficiency was demonstrated on recombinant 3-dehydroshikimate dehydratase (DHSase or wtAsbF) from Bacillus thuringiensis serovar konkukian 97-27 (hereafter, B. thuringiensis 97-27). The wtAsbF is relatively unstable at 37 °C, in vitro (t1/237 = 15 min), in the absence of divalent metal. We adopted a structure-based design to identify stabilizing mutations and created a combinatorial library based upon predicted mutations at specific locations on the enzyme surface. A diversified asbF library (∼2000 variants) was expressed in E. coli harboring a green fluorescent protein (GFP) reporter system linked to the product of wtAsbF activity (3,4-dihydroxybenzoate, DHB). Mutations detrimental to DHSase function were rapidly eliminated using a high throughput fluorescence activated cell sorting (FACS) approach. After a single sorting round and heat screen at 50 °C, a triple AsbF mutant (Mut1), T61N, H135Y, and H257P, was isolated and characterized. The half-life of Mut1 at 37 °C was >10-fold higher than the wtAsbF (t1/237 = 169 min). Further, the second-order rate constants for both wtAsbF and Mut1 were approximately equal (9.9 × 105 M-1 s-1, 7.8 × 105 M-1 s-1, respectively), thus demonstrating protein thermostability did not come at the expense of enzyme thermophilicity. In addition, in vivo overexpression of Mut1 in E. coli resulted in a ∼60-fold increase in functional enzyme when compared to the wild-type enzyme under the identical expression conditions. Finally, overexpression of the thermostable AsbF resulted in an approximate 80-120% increase in DHB accumulation in the media relative to the wild-type enzyme.


With the help of kin? Household composition and reproduction in the Netherlands, 1842-1920.
Relatives play an important role in human reproduction according to evolutionary theories of reproductive behavior, but previous empirical studies show large differences in the effects of kin on fertility outcomes. In our paper we examine the effect of co-resident kin and non-kin on the length of birth intervals over the reproductive life course of Dutch women born between 1842 and 1920. We estimate Cox proportional hazard models for parity progression based on the presence of kin and non-kin in the household while controlling for a large number of individual and community-level characteristics. We find that couples living with their brothers experienced shorter birth intervals whereas couples residing with a widowed father had relatively longer birth intervals. The effects of these types of kin on reproduction were most pronounced up to the birth of the fifth child, but not thereafter. We found no effect for mothers or other types of kin.


[Reflex therapy of vertebrogenic radicular pain syndromes].
The authors analyse results of the treatment of 120 patients with vertebrogenic radicular pain syndromes by reflexotherapy, electropuncture, electroacupuncture and acupuncture, applied both alone and combined under conditions of a neurological department. Examination of the patients included a comprehensive analysis of neurological symptomatology and its variations in the time course of the treatment exercised according to a specially elaborated schedule of case history. Approaches to the choice of the most effective methods of reflexotherapy, depending on the extent of injury, intensity of the pain syndrome and the state of the vegetative nervous system are offered and the efficacy of the methods of reflexotherapy is clinically compared.


Effects of CO2 and temperature on photosynthetic performance in the diatom Chaetoceros gracilis.
CO2 concentration and temperature for growth of photosynthetic organisms are two important factors to ensure better photosynthetic performance. In this study, we investigated the effects of CO2 concentration and temperature on the photosynthetic performance in a marine centric diatom Chaetoceros gracilis. Cells were grown under four different conditions, namely, at 25 °C with air bubbling, at 25 °C with a supplementation of 3% CO2, at 30 °C with air bubbling, and at 30 °C with the CO2 supplementation. It was found that the growth rate of cells at 30 °C with the CO2 supplementation is faster than those at other three conditions. The pigment compositions of cells grown under the different conditions are altered, and fluorescence spectra measured at 77 K also showed different peak positions. A novel fucoxanthin chlorophyll a/c-binding protein complex is observed in the cells grown at 30 °C with the CO2 supplementation but not in the other three types of cells. Since oxygen-evolving activities of the four types of cells are almost unchanged, it is suggested that the CO2 supplementation and growth temperature are involved in the regulation of photosynthetic light-harvesting apparatus in C. gracilis at different degrees. Based on these observations, we discuss the favorable growth conditions for C. gracilis.


The Divergence of Chromosome Structures and 45S Ribosomal DNA Organization in Cucumis debilis Inferred by Comparative Molecular Cytogenetic Mapping.
Cucumis debilis W.J.de Wilde & Duyfjes is an annual and monoecious plant. This species is endemic to Southeast Asia, particularly Vietnam. However, C. debilis is rarely studied, and no detailed information is available regarding its basic chromosome number, 45S ribosomal DNA (rDNA) status, and divergence among other Cucumis species. In this study, we characterized the morphological characters and determined and investigated the basic chromosome number and chromosomal distribution of 45S rDNA of C. debilis using the fluorescent in situ hybridization (FISH) technique. A maximum likelihood tree was constructed by combining the chloroplast and internal transcribed spacer of 45S rDNAs to infer its relationship within Cucumis. C. debilis had an oval fruit shape, green fruit peel, and protrusion-like white spots during the immature fruit stage. FISH analysis using 45S rDNA probe showed three pairs of 45S rDNA loci located at the terminal region in C. debilis, similar to C. hystrix. Meanwhile, two, two, and five pairs of 45S rDNA loci were observed for C. melo, C. metuliferus, and C. sativus, respectively. One melon (P90) and cucumber accessions exhibited different chromosomal localizations compared with other members of Cucumis. The majority of Cucumis species showed the terminal location of 45S rDNA, but melon P90 and cucumber exhibited terminal-interstitial and all interstitial orientations of 45S rDNA loci. Based on molecular cytogenetics and phylogenetic evidence, C. debilis is more closely related to cucumber than melon. Therefore, C. debilis may serve as a potential parental accession for genetic improvement of cucumber through interspecific hybridization.


Asymmetric alpha-alkylation of N'-tert-butanesulfinyl amidines. Application to the total synthesis of (6R,7S)-7-amino-7,8-dihydro-alpha-bisabolene.
A highly diastereoselective alpha-alkylation of N'-tert-butanesulfinyl amidines has been developed along with methods for converting the alkylation products to enantiomerically enriched amines that incorporate both alpha- and beta-stereocenters. The utility of this chemistry is further demonstrated by the first asymmetric synthesis of the antimicrobial marine natural product (6R,7S)-7-amino-7,8-dihydro-alpha-bisabolene.


[Informed Consent in the Humanization of the Cesarean: A Preliminary Study].
The informed consent (IC) is a process based on dialogue between the professional and the patient in which he freely decides on possible interventions in their health. This is applicable to caesarean delivery and if it meets a number of conditions will help to improve the process of ″humanization″ of birth. The overall objective of this study is to analyze preliminarily in several hospitals in the Region of Murcia the IC in caesarean delivery. To this end, we have revised the documents of IC and we studied who, where, when and how the IC process is done. The results show that all hospitals are based on the same document, and although the documents take into account all the elements of a IC, do not indicate the date of their design or subsequent revisions. It does not contemplate the risks and complications that caesarean section can have on the newborn, mother, and mother-child relationship later. It is noted that the document of IC normally is delivers by gynecologist in the consultation, when intervention is programmed, although it are sometimes nurses, who after admission to the hospital give it to sign the patient. In urgent caesarean sections, there are some hospitals that in life-threatening situation, do not offer the document of IC to women. In others, it is offered hastily by the gynecologist or midwife. In conclusion, the IC is a process which used correctly, favors the relationship between women and health professionals in the intervention of cesarean section. Although this process and the documents of IC examined in our study, have presented many positive aspects, the humanization of caesarean could be increased improving with the preparation and updating of these documents and coordinating the various professionals.