Microbiological analysis of environmental samples collected from child care facilities in North and South Carolina.
Children cared for outside the home are at an increased risk of enteric disease. Microbiological analyses were performed on environmental samples collected from child care facilities in North and South Carolina. There were 326 samples collected from 40 facilities corresponding to common surfaces (77% of samples) and the hands of care providers (23% of samples). Samples were analyzed for total aerobic plate counts (APCs), total coliforms, biotype I Escherichia coli, and pathogens Shigella spp, Salmonella enterica, E coli O157, Campylobacter jejuni, and human norovirus. Median APCs and coliform counts for hands were 4.6 and 1.0 log10 colony-forming units (CFU) per hand, respectively. Median APCs for surfaces were 2.0 and 2.6 log10 CFU for flat and irregular surfaces, respectively. Coliforms were detected in 16% of samples, with counts ranging from 1.0 log10 to > 4.3 log10 CFU, with higher counts most often observed for hand rinse samples. Biotype I E coli counts were below assay detection limits (< 1 log10 CFU) for all but 1 sample. No samples were positive for any of the 4 bacterial pathogens, whereas 4 samples showed evidence of human norovirus RNA. The relative absence of pathogens and biotype I E coli in environmental samples suggests the child care facilities sampled in this study managed fecal contamination well.


Therapeutic benefits of Indole-3-Carbinol in adjuvant-induced arthritis and its protective effect against methotrexate induced-hepatic toxicity.
Rheumatoid arthritis (RA) being an incapacitating disease requires early effective intervention. Considering Methotrexate (MTX)- the first line of treatment for RA- intoxicates the liver; therefore, alternative therapies with similar efficacy yet lower cytotoxicity are desired. Indole-3-Carbinol (I3C) which is found in cruciferous vegetables was examined for its possible therapeutic potentials in comparison with MTX by investigating its anti-inflammatory, anti-arthritic, anti-oxidant, and hepatoprotective potentials in adjuvant-induced arthritis (AIA) rat model. Arthritis was induced in Sprague Dawley rats by injection of Complete Freund's Adjuvant (CFA). Arthritic rats were treated with I3C and/or MTX. To examine the anti-inflammatory and anti-arthritic effect, the following parameters were assessed: body weight, macroscopic scoring of the hind paw, the level of the pivotal cytokines (TNF-α, IL-6) heavily involved in the pathogenesis, spleen index, and erythrocyte sedimentation rate. At a histological level, the tibiotarsal joint was stained with several specific stains. To assess the hepatoprotective and anti-oxidant effects, several oxidative stress parameters were monitored, and the liver histology was examined. Both I3C and MTX attenuated the inflammation that was aggravated by arthritis by downregulating the inflammatory markers and mediators and alleviating the histopathological changes affecting the tibiotarsal joint. I3C attenuated the liver impairment that was initiated by arthritis and MTX treatment. It did so by downregulating the pro-oxidants and up-regulating the anti-oxidant defenses and by reducing the pathological changes affecting the liver. Our results suggest that I3C is as potent as MTX as an anti-inflammatory and anti-arthritic agent. In addition, I3C does so while protecting the liver from damage as opposed to MTX.


Ethics of Infection Control Measures for Carriers of Antimicrobial Drug-Resistant Organisms.
Many countries have implemented infection control measures directed at carriers of multidrug-resistant organisms. To explore the ethical implications of these measures, we analyzed 227 consultations about multidrug resistance and compared them with the literature on communicable disease in general. We found that control measures aimed at carriers have a range of negative implications. Although moral dilemmas seem similar to those encountered while implementing control measures for other infectious diseases, 4 distinct features stand out for carriage of multidrug-resistant organisms: carriage presents itself as a state of being; carriage has limited relevance for the health of the carrier; carriage has little relevance outside healthcare settings; and antimicrobial resistance is a slowly evolving threat on which individual carriers have limited effect. These features are of ethical relevance because they influence the way we traditionally think about infectious disease control and urge us to pay more attention to the personal experience of the individual carrier.


Generalizability of Human Activity Recognition Machine Learning Models from non-Parkinson's to Parkinson's Disease Patients.
Recent evidence shows that high-intensity exercises reduce tremors and stiffness in Parkinson's disease (PD). However, there is insufficient evidence on the types of exercises; in effect, high-intensity may be a personalized measure. Recent progress in automated Human Activity Recognition using machine learning (ML) models shows potential for better monitoring of PD patients. However, ML models must be calibrated to ignore tremors and accurately identify activity and its intensity. We report findings from a study where we trained ML models using data from medically validated triple synchronous sensors connected to 8 non-PD subjects performing 32 exercises. We then tested the models to identify exercises performed by 8 PD patients at different stages of the disease. Our analysis shows that better data preprocessing before modeling can provide some model generalizability. However, it is extremely challenging, as the models work with high accuracy on one group (Healthy or PD patients) (F1=0.88-0.94) but not on both groups.Clinical relevance-Patients with Parkinson's and other motor-generative diseases can now accurately measure physical activity with machine learning approaches. Clinicians, caregivers, and apps can make accurate, personalized exercise recommendations to augment medications that reduce tremors and stiffness.


Effect of nanovaccine chemistry on humoral immune response kinetics and maturation.
Acute respiratory infections represent a significant portion of global morbidity and mortality annually. There is a critical need for efficacious vaccines against respiratory pathogens. To vaccinate against respiratory disease, pulmonary delivery is an attractive route because it mimics the route of natural infection and can confer both mucosal and systemic immunity. We have previously demonstrated that a single dose, intranasal vaccine based on polyanhydride nanoparticles elicited a protective immune response against Yersinia pestis for at least 40 weeks after immunization with F1-V. Herein, we investigate the effect of nanoparticle chemistry and its attributes on the kinetics and maturation of the antigen-specific serum antibody response. We demonstrate that manipulation of polyanhydride nanoparticle chemistry facilitated differential kinetics of development of antibody titers, avidity, and epitope specificity. The results provide new insights into the underlying role(s) of nanoparticle chemistry in providing long-lived humoral immunity and aid in the rational design of nanovaccine formulations to induce long-lasting and mature antibody responses.


Injuries of the pelvic ureter.
Ureteral injuries may cause infection, fistula, and even death. By revealing abnormalities, preoperative pyelography warns the surgeon of hazards to be anticipated. Injury found during operation is managed by reimplantation, reanastomosis, or ureterostomy in situ, while deliberate ligature is avoided. Postoperative ureteral fistula is treated by reimplantation, although occasionally a tongue of reflected bladder wall converted into a tube is required. In the absence of skilled urologic advice, ureteral injuries may be managed by the gynecologist.


Shear stress regulates endothelial nitric-oxide synthase promoter activity through nuclear factor kappaB binding.
We have previously demonstrated that shear stress increases transcription of the endothelial nitric-oxide synthase (eNOS) by a pathway involving activation of the tyrosine kinase c-Src and extracellular signal-related kinase 1/2 (ERK1/2). In the present study sought to determine the events downstream of this pathway. Shear stress activated a human eNOS promoter chloramphenicol acetyl-CoA transferase chimeric construct in a time-dependent fashion, and this could be prevented by inhibition of the c-Src and MEK1/2. Studies using electromobility shift assays, promoter deletions, and promoter mutations revealed that shear activation of the eNOS promoter was due to binding of nuclear factor kappaB subunits p50 and p65 to a GAGACC sequence -990 to -984 base pairs upstream of the eNOS transcription start site. Shear induced nuclear translocation of p50 and p65, and activation of the eNOS promoter by shear could be prevented by co-transfection with a dominant negative I kappa Balpha. Exposure of endothelial cells to shear resulted in Ikappa kinase phosphorylation, and this was blocked by the MEK1/2 inhibitor PD98059 and the cSrc inhibitor PP1, suggesting these signaling molecules are upstream of NFkappaB activation. These experiments indicate that shear stress increases eNOS transcription by NFkappaB activation and p50/p65 binding to a GAGACC sequence present of the human eNOS promoter. While NFkappaB activation is generally viewed as a proinflammatory stimulus, the current data indicate that its transient activation by shear may increase expression of eNOS, which via production of nitric oxide could convey anti-inflammatory and anti-atherosclerotic properties.


Preparation and chemical composition of the cell walls of mature infectious dense forms of meningopneumonitis organisms.
Relatively large-scale production and purification of meningopneumonitis organisms was developed for chemical and immunological studies on cell walls of the infectious dense forms. By disruption of purified organisms with glass beads in a Mickle shaker, highly purified preparations of cell walls were obtained by sucrose density gradient centrifugation, enzyme digestion, and sodium dodecyl sulfate treatment. The dry-weight recovery of purified cell walls from intact organisms was about 13%. When (32)P-labeled preparations of cell walls were fractionated into acid-soluble, lipid, ribonucleic acid (RNA), deoxyribonucleic (DNA), and residual fractions, about 80% of the (32)P in cell wall preparations was recovered in the phospholipid fraction, which corresponded to about 3% of the total phospholipid in the intact organisms. About 7% of the (32)P in purified cell walls was recovered in the RNA and DNA fractions respectively, but this corresponds to only about 0.4% of the (32)P found in those fractions in intact organisms. From dry-weight determinations, it was calculated that the purified cell wall preparations contained only 0.6% total nucleic acids, and these are probably not true cell wall constituents. These cell walls contained 70 to 75% protein, corresponding to about 14% of the protein in intact organisms. Amino acid analysis of these protein showed the existence of all common amino acids, glucosamine, and galactosamine. However, no muramic acid was detected by the methods employed.


Reduction of excessive unfolded protein response by 4-phenylbutyric acid may mitigate procymidone-induced testicular damage in mice by changing the levels of circRNA Scar and circZc3h4.
Procymidone (PCM) exposure below the no-observed-effect level triggers changes in circRNA Scar and circZc3h4 and overactivation of the unfolded protein response (UPR) in mice, culminating in testicular injury. The 4-phenyl butyric acid (4-PBA) is known to stabilize proteins and reduce the UPR. This study employed an in vitro system in which mouse testes were cultured with 1 × 10-5 M PCM and varying concentrations (0, 20, 40, and 80 mM) of 4-PBA; 4-week-old male mice were subsequently treated with 100 mg/kg/d PCM (suspended in corn oil) and/or 100 mg/kg/d 4-PBA for 21 d, consecutively. The treatments were as follows: the negative control (NC) group was orally administered corn oil; the positive control (PC) group was orally administered PCM; the 4-PBA group was intraperitoneally injected with 4-PBA; the 4-PBA-I group was orally administered PCM and 4-PBA simultaneously; the 4-PBA-II group received daily administration of 4-PBA 24 h prior to PCM; and the 4-PBA-III group was intraperitoneally injected with 4-PBA for 7 d after 21 d of PCM administration. However, the 4-PBA intervention groups showed no considerable changes in the overall or testicular appearance of mice. In vitro, 4-PBA inhibited the PCM-induced testicular injury, with the most significant effect observed at 80 mM. In vivo, the 4-PBA-III group exhibited the best in vivo effects. Our findings indicate that 4-PBA conferred testicular protection by decreasing PCM-induced circRNA Scar, elevating circZc3h4, and suppressing UPR both in vitro and in vivo. It has been hypothesized that 4-PBA mitigates testicular damage by reducing excessive UPR levels.


Secondary leukemia in a child with neuroblastoma while on oral etoposide: what is the cause?
To date little has been reported about the risk of therapy-related leukaemia (t-AML) in children receiving oral etoposide therapy. The authors present a case of t-AML that developed in a child with metastatic neuroblastoma 18 months after he received oral etoposide, given for palliation purpose. The leukemic blasts were examined by morphological, immunohistochemical, cytogenetic, and molecular genetic analyses. Although the t-AML developed following oral etoposide therapy, the child had previously received high-dose, multiagent chemotherapy, and rearrangement of the MLL gene was not demonstrated. The use of modern multiagent therapy often makes it difficult to appropriately apportion blame for causation of specific side effects. Moreover, the etiology of t-AML and mechanism of leukemogenesis are likely to be multifactorial and complex. Further studies on the precise association with different therapies are thus needed. Oral etoposide remains an effective palliative agent and its usage should not be excluded without most careful consideration of the risks.


Childhood adversities and adult psychiatric disorders in the national comorbidity survey replication I: associations with first onset of DSM-IV disorders.
Although significant associations of childhood adversities (CAs) with adult mental disorders have been documented consistently in epidemiological surveys, these studies generally have examined only 1 CA per study. Because CAs are highly clustered, this approach results in overestimating the importance of individual CAs. Multivariate CA studies have been based on insufficiently complex models. To examine the joint associations of 12 retrospectively reported CAs with the first onset of DSM-IV disorders in the National Comorbidity Survey Replication using substantively complex multivariate models. Cross-sectional community survey with retrospective reports of CAs and lifetime DSM-IV disorders. Household population in the United States. Nationally representative sample of 9282 adults. Lifetime prevalences of 20 DSM-IV anxiety, mood, disruptive behavior, and substance use disorders assessed using the Composite International Diagnostic Interview. The CAs studied were highly prevalent and intercorrelated. The CAs in a maladaptive family functioning (MFF) cluster (parental mental illness, substance abuse disorder, and criminality; family violence; physical abuse; sexual abuse; and neglect) were the strongest correlates of disorder onset. The best-fitting model included terms for each type of CA, number of MFF CAs, and number of other CAs. Multiple MFF CAs had significant subadditive associations with disorder onset. Little specificity was found for particular CAs with particular disorders. Associations declined in magnitude with life course stage and number of previous lifetime disorders but increased with length of recall. Simulations suggest that CAs are associated with 44.6% of all childhood-onset disorders and with 25.9% to 32.0% of later-onset disorders. The fact that associations increased with length of recall raises the possibility of recall bias inflating estimates. Even considering this, the results suggest that CAs have powerful and often subadditive associations with the onset of many types of largely primary mental disorders throughout the life course.


The Botanic Mission to Mozambique (1942-1948): contributions to knowledge of the medicinal flora of Mozambique.
This article reviews the historical and scientific findings of the Botanic Mission to Mozambique (1942-1948) under the Tropical Botanic Garden of the Instituto de Investigação Científica Tropical, in Lisbon, highlighting the collectors' field notes with the aim of identifying the traditional medicinal uses of Mozambican flora. Having collated information on 71 taxa (70 species and one genus), the medicinal usage of 34 species presumably not yet reported in Mozambique was identified, including five whose therapeutic use still had not yet been described in the African continent. Overall, 58 uses presumably not yet reported in Mozambique were recorded.


Purification and properties of phosphoprotein phosphatase from rabbit heart.
A low molecular weight phosphoprotein phosphatase acting on muscle phosphorylase a has been purified to homogeneity from rabbit heart by acid precipitation, ethanol treatment, and chromatography on Sephadex G-75 and Sepharose-histone. The purified enzyme showed a single band when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; the molecular weight calculated by this method was 34,000. The S20,w value and Stockes radius for the enzyme were 3.45 and 24.0 A, respectively. Using these two values, a molecular weight of 35,000 was calculated. Purified enzyme showed a wide substrate specificity and catalyzed the dephosphorylation of phosphorylase a, glycogen synthase D, phosphorylated histone, and phosphorylated casein. A heat-stable protein inhibitor for this enzyme with phosphorylase a as the substrate was also shown to be present in crude extracts of rabbit heart. It inhibited the dephosphorylation of phosphorylase a by phosphoprotein phosphatase by decreasing the Vmax of the reaction.


The impact of chromatin organization of vitamin D target genes.
The vitamin D receptor (VDR), the nuclear receptor for 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3), controls gene expression by binding discrete DNA sequences in promoter regions of target genes, referred to as 1alpha,25(OH)2D3 response elements (VDREs). Although these elements are well characterized in vitro, the function of VDREs in living cells in the context of chromatin is still largely unknown. To resolve this issue, 7 to 8 kB of the promoter regions of the primary 1alpha,25(OH)2D3 target genes CYP24, cyclin C and p21(Waf1/Cip1) were studied by chromatin immunoprecipitation (ChIP) assays using antibodies against acetylated histone H4 (to assess the global chromatin status) and various other components of VDR-dependent gene activation, such as VDR, retinoid X receptor (RXR), coactivator (CoA) and corepressor proteins. This approach identified three to four functional VDREs per gene promoter. In parallel, the extended analysis of the gene areas, of all six members of the insulin-like growth factor binding protein (IGFBP) family (i.e., 10 kB of promoter, introns, exons and 10 kB of the downstream region) were screened in silico for putative VDR-binding sites. Gel shift, reporter gene and ChIP assays identified, in total, ten functional VDREs in the genes IGFBP1, 3 and 5 and real-time PCR confirmed that these genes are primary VDR targets. Taken together, these results suggest that a reasonable proportion of all VDR target genes, if not all, are under the control of multiple VDREs. These results will have an impact on the development of therapeutic regimes for diseases, such as cancer, that use 1alpha,25(OH)2D3 and its analogs.


The direct inhibitory effect of dutasteride or finasteride on androgen receptor activity is cell line specific.
Finasteride and dutasteride were developed originally as 5α-reductase inhibitors to block the conversion of testosterone to dihydrotestosterone (DHT). These drugs may possess off-target effects on the androgen receptor (AR) due to their structural similarity to DHT. A total of four human prostate cancer cell models were examined: LNCaP (T877A mutant AR), 22Rv1 (H874Y mutant AR), LAPC4 (wild-type AR), and VCaP (wild-type AR). Cells were cultured in 10% charcoal-stripped fetal bovine serum, either with or without DHT added to the medium. AR activity was evaluated using the ARE-luciferase assay or the expression of AR regulated genes. Dutasteride was more potent than finasteride in interfering with DHT-stimulated AR signaling. Disruption of AR function was accompanied by decreased cell growth. Cells that rely on DHT for protection against death were particularly vulnerable to dutasteride. Different prostate cancer cell models exhibited different sensitivities to dutasteride and finasteride. LNCaP was most sensitive, LAPC4 and VCaP were intermediate, while 22Rv1 was least sensitive. Regardless of the AR genotype, if AR was transfected into drug-sensitive cells, AR was inhibited by drug treatment; and if AR was transfected into drug-resistant cells, AR was not inhibited. The direct inhibitory effect of dutasteride or finasteride on AR signaling is cell line specific. Mutations in the ligand binding domain of AR do not appear to play a significant role in influencing the AR antagonistic effect of these drugs. Subcellular constituent is an important factor in determining the drug effect on AR function.


[Immediate effect of nasal continuous positive pressure in the treatment of obstructive sleep apnea].
In 8 patients with severe sleep apnea syndrome we have studied the immediate effect of nasal continuous positive airway pressure. A good adaptation to the machine during their stay at the laboratory and the beneficial immediate effect on somnolence was observed. There was found a better efficiency of sleep, shortening of latency, decrease of stage 1, increase of REM and shortening of the latency of REM (rebound of REM sleep). Significant correlations were found between the needed pressure to eliminate apneas and the proportion of stage 1 (r = 0.83), of REM stage (r = 0.72) and stage 2 (r = -0.84) of basal night sleep.


Regulation of gene expression by retinoids.
Vitamin A serves as substrate for the biosynthesis of several derivates (retinoids) which are important for cell growth and cell differentiation as well as for vision. Retinoic acid is the major physiologically active form of vitamin A regulating the expression of different genes. At present, hundreds of genes are known to be regulated by retinoic acid. This regulation is very complex and is, in turn, regulated on many levels. To date, two families of retinoid nuclear receptors have been identified: retinoic acid receptors and retinoid X receptors, which are members of the steroid hormone receptor superfamily of ligand-activated transcription factors. In order to regulate gene expression, all-trans retinal needs to be oxidized to retinoic acid. All-trans retinal, in turn, can be produced during oxidation of all-trans retinol or in a retinol-independent metabolic pathway through cleavage of β-carotene with all-trans retinal as an intermediate metabolite. Recently it has been shown that not only retinoic acid is an active form of vitamin A, but also that all-trans retinal can play an important role in gene regulation. In this review we comprehensively summarize recent literature on regulation of gene expression by retinoids, biochemistry of retinoid receptors, and molecular mechanisms of retinoid-mediated effects on gene regulation.


Interaction between the skeletal muscle type 1 Na+ channel promoter E-box and an upstream repressor element. Release of repression by myogenin.
We have defined how four elements that regulate expression of the rat skeletal muscle type 1 sodium channel (SkM1) gene cooperate to yield specific expression in differentiated muscle. A basal promoter region containing within it a promoter E-box (-31/-26) is broadly expressed in many cells, including myoblasts and myotubes; mutations within the promoter E-box that disrupt binding of the myogenic basic helix-loop-helix (bHLH) factors reduce expression in all cell types only slightly. Sequential addition of upstream elements to the wild-type promoter confer increasing specificity of expression in differentiated cells, even though all three upstream elements, including a positive element (-85/-57), a repressor E-box (-90/-85), and upstream repressor sequences (-135/-95), bind ubiquitously expressed transcription factors. Mutations in the promoter E-box that disrupt the binding of the bHLH factors counteract the specificity conferred by addition of the upstream elements, with the greatest interaction observed between the upstream repressor sequences and the promoter E-box. Forced expression of myogenin in myoblasts releases repression exerted by the upstream repressor sequences in conjunction with the wild-type, but not mutant, promoter E-box, and also initiates expression of the endogenous SkM1 protein. Our data suggest that particular myogenic bHLH proteins bound at the promoter E-box control expression of SkM1 by releasing repression exerted by upstream repressor sequences in differentiated muscle cells.


The Molecular Mechanism of Antioxidation of Huolisu Oral Liquid Based on Serum Analysis and Network Analysis.
Huolisu Oral Liquid (HLS), a well-known traditional Chinese medicine (TCM) prescription, is an over-the-counter drug that is registered and approved by the State Food and Drug Administration (Approval No. Z51020381). HLS has been widely applied in the clinical treatment of cognitive disorders and has effects on delaying aging. The antioxidant effects of HLS are closely related to its antiaging activities, but the underlying mechanisms are unclear. In this study, the potential antioxidant ingredients of HLS were screened based on serum pharmacochemistry and network pharmacology, and the potential mechanisms involved in HLS antioxidant effects were preliminarily explored. Further, the antioxidant effects of HLS were verified by in vivo and in vitro experiments. The results showed that potential antioxidant ingredients could affect the toxic advanced glycation end products-receptor for advanced glycation end products (TAGE-RAGE) signaling, mitogen-activated protein kinase (MAPK) signaling, interleukin (IL)-17 signaling, tumor necrosis factor (TNF) signaling, toll-like receptors (TLRs), cyclic adenosine monophosphate (cAMP) signaling, hypoxia-inducible factor (HIF)-1 signaling, and other related pathways by regulating GAPDH, AKT1, TP53, MAPK1, JUN, and other associated targets. Thus, HLS may reduce inflammation, control the release of inflammatory cytokines, and regulate mitochondrial autophagy and metabolic abnormalities to ultimately play an antioxidant role. This is the first study attempting to construct a multilevel network of "HLS-antioxidant targets" based on serum pharmacochemistry and network pharmacology to explore the relationship between HLS and antioxidation and the molecular mechanisms of antioxidation combined with bioinformatics functional analysis and lays a foundation for further elucidating the antioxidant mechanisms of HLS.


Cardiorespiratory fitness and adiposity in metabolically healthy overweight and obese youth.
Controversy exists surrounding the contribution of fitness and adiposity as determinants of the Metabolically Healthy Overweight(MHO) phenotype in youth. This study investigated the independent contribution of cardiorespiratory fitness and adiposity to the MHO phenotype among overweight and obese youth. This cross-sectional study included 108 overweight and obese youth classified as MHO (no cardiometabolic risk factors) or non-MHO (≥1 cardiometabolic risk factor), based on age- and gender specific cut-points for fasting glucose, triglycerides, high-density lipoprotein cholesterol, systolic and diastolic blood pressure, and hepaticsteatosis. Twenty-five percent of overweight and obese youth were classified as MHO. This phenotype was associated with lower BMIz-score (BMI z-score: 1.8 ± 0.3 vs 2.1 ± 0.4, P = .02) and waist circumference (99.7 ± 13.2 vs 106.1 ± 13.7 cm, P = .04) compared with non-MHO youth. When matched for fitness level and stratified by BMI z-score (1.6 ± 0.3 vs 2.4 ± 0.2), the prevalence of MHO was four fold higher in the low BMI z-score group (27% vs 7%; P = .03).Multiple logistic regression analyses revealed that the best predictor of MHO was the absence of hepatic steatosis even after adjusting for waist circumference (odds ratio 0.57, 95% confidence interval 0.40–0.80) or BMI z-score (odds ratio 0.59, 95% confidence interval 0.43–0.80). The MHO phenotype was present in 25% of overweight and obese youth and is strongly associated with lower levels of adiposity,and the absence of hepatic steatosis, but not with cardiorespiratory fitness.


Genomic analysis for managing small and endangered populations: a case study in Tyrol Grey cattle.
Analysis of genomic data is increasingly becoming part of the livestock industry. Therefore, the routine collection of genomic information would be an invaluable resource for effective management of breeding programs in small, endangered populations. The objective of the paper was to demonstrate how genomic data could be used to analyse (1) linkage disequlibrium (LD), LD decay and the effective population size (NeLD); (2) Inbreeding level and effective population size (NeROH) based on runs of homozygosity (ROH); (3) Prediction of genomic breeding values (GEBV) using small within-breed and genomic information from other breeds. The Tyrol Grey population was used as an example, with the goal to highlight the potential of genomic analyses for small breeds. In addition to our own results we discuss additional use of genomics to assess relatedness, admixture proportions, and inheritance of harmful variants. The example data set consisted of 218 Tyrol Grey bull genotypes, which were all available AI bulls in the population. After standard quality control restrictions 34,581 SNPs remained for the analysis. A separate quality control was applied to determine ROH levels based on Illumina GenCall and Illumina GenTrain scores, resulting into 211 bulls and 33,604 SNPs. LD was computed as the squared correlation coefficient between SNPs within a 10 mega base pair (Mb) region. ROHs were derived based on regions covering at least 4, 8, and 16 Mb, suggesting that animals had common ancestors approximately 12, 6, and 3 generations ago, respectively. The corresponding mean inbreeding coefficients (F ROH) were 4.0% for 4 Mb, 2.9% for 8 Mb and 1.6% for 16 Mb runs. With an average generation interval of 5.66 years, estimated NeROH was 125 (NeROH>16 Mb), 186 (NeROH>8 Mb) and 370 (NeROH>4 Mb) indicating strict avoidance of close inbreeding in the population. The LD was used as an alternative method to infer the population history and the Ne. The results show a continuous decrease in NeLD, to 780, 120, and 80 for 100, 10, and 5 generations ago, respectively. Genomic selection was developed for and is working well in large breeds. The same methodology was applied in Tyrol Grey cattle, using different reference populations. Contrary to the expectations, the accuracy of GEBVs with very small within breed reference populations were very high, between 0.13-0.91 and 0.12-0.63, when estimated breeding values and deregressed breeding values were used as pseudo-phenotypes, respectively. Subsequent analyses confirmed the high accuracies being a consequence of low reliabilities of pseudo-phenotypes in the validation set, thus being heavily influenced by parent averages. Multi-breed and across breed reference sets gave inconsistent and lower accuracies. Genomic information may have a crucial role in management of small breeds, even if its primary usage differs from that of large breeds. It allows to assess relatedness between individuals, trends in inbreeding and to take decisions accordingly. These decisions would be based on the real genome architecture, rather than conventional pedigree information, which can be missing or incomplete. We strongly suggest the routine genotyping of all individuals that belong to a small breed in order to facilitate the effective management of endangered livestock populations.


Whole-genome microarrays of fission yeast: characteristics, accuracy, reproducibility, and processing of array data.
The genome of the fission yeast Schizosaccharomyces pombe has recently been sequenced, setting the stage for the post-genomic era of this increasingly popular model organism. We have built fission yeast microarrays, optimised protocols to improve array performance, and carried out experiments to assess various characteristics of microarrays. We designed PCR primers to amplify specific probes (180-500 bp) for all known and predicted fission yeast genes, which are printed in duplicate onto separate regions of glass slides together with control elements (approximately 13,000 spots/slide). Fluorescence signal intensities depended on the size and intragenic position of the array elements, whereas the signal ratios were largely independent of element properties. Only the coding strand is covalently linked to the slides, and our array elements can discriminate transcriptional direction. The microarrays can distinguish sequences with up to 70% identity, above which cross-hybridisation contributes to the signal intensity. We tested the accuracy of signal ratios and measured the reproducibility of array data caused by biological and technical factors. Because the technical variability is lower, it is best to use samples prepared from independent biological experiments to obtain repeated measurements with swapping of fluorochromes to prevent dye bias. We also developed a script that discards unreliable data and performs a normalization to correct spatial artefacts. This paper provides data for several microarray properties that are rarely measured. The results define critical parameters for microarray design and experiments and provide a framework to optimise and interpret array data. Our arrays give reproducible and accurate expression ratios with high sensitivity. The scripts for primer design and initial data processing as well as primer sequences and detailed protocols are available from our website.


Repair of congenitally dysplastic aortic valve by bicuspidization: midterm results.
Repair of congenital aortic valve (CAV) lesions may be achieved by creation of either tricuspid or bicuspid valve morphology. To define feasibility and outcome of CAV repair by bicuspidization a 10-year experience was reviewed. Between January 2002 and December 2011, 147 consecutive patients underwent operation for CAV insufficiency; 58 had valve or root repair (group 1) and 89 had valve or root replacement (group 2). Patients having repair were younger (42.9 vs 51.3 years, p=0.001), with lesser prevalence of severe insufficiency (72% vs 90%, p=0.002). In patients having repair, morphology of CAV was bicuspid in 51, monocuspid in 4, and quadricuspid in 3, whereas in the replacement group it was bicuspid in 87 and quadricuspid in 2 (p=0.04). Surgery consisted of an isolated aortic valve procedure in 20 versus 45 patients, associated with aortic root or ascending aortic repair in 38 versus 44 patients, in group 1 versus 2 (p=0.04). There were no hospital and 3 late deaths during a mean follow-up of 3.8±2.5 years (range 0.2 to 10.0). Eight-year survival (89%±10% vs 97%±2% [p=0.7]), freedom from valve-related events (84%±10% vs 89%±4% [p=0.8]), and freedom from aortic valve reoperation (95%±3% vs 93%±3% [p=0.6]) were comparable. Risk factors for reoperation at univariate analysis were isolated valve surgery (p=0.001), Ross operation (p=0.001), and endocarditis (p=0.002). Follow-up echocardiography of repair patients showed mild or less aortic insufficiency in 51 (88%) and mild or less stenosis in 57 (98%). Valve repair by preservation or creation of bicuspid morphology is feasible in almost half of all comers with CAV insufficiency, with satisfactory and stable midterm functional outcome. Rates of valve-related adverse events and reoperation are similar to those of patients having replacement.


The multiform motor cortical output: Kinematic, predictive and response coding.
Observing actions performed by others entails a subliminal activation of primary motor cortex reflecting the components encoded in the observed action. One of the most debated issues concerns the role of this output: Is it a mere replica of the incoming flow of information (kinematic coding), is it oriented to anticipate the forthcoming events (predictive coding) or is it aimed at responding in a suitable fashion to the actions of others (response coding)? The aim of the present study was to disentangle the relative contribution of these three levels and unify them into an integrated view of cortical motor coding. We combined transcranial magnetic stimulation (TMS) and electromyography recordings at different timings to probe the excitability of corticospinal projections to upper and lower limb muscles of participants observing a soccer player performing: (i) a penalty kick straight in their direction and then coming to a full stop, (ii) a penalty kick straight in their direction and then continuing to run, (iii) a penalty kick to the side and then continuing to run. The results show a modulation of the observer's corticospinal excitability in different effectors at different times reflecting a multiplicity of motor coding. The internal replica of the observed action, the predictive activation, and the adaptive integration of congruent and non-congruent responses to the actions of others can coexist in a not mutually exclusive way. Such a view offers reconciliation among different (and apparently divergent) frameworks in action observation literature, and will promote a more complete and integrated understanding of recent findings on motor simulation, motor resonance and automatic imitation.


Current and upcoming point-of-care diagnostics for schistosomiasis.
Point-of-care (POC) diagnostics are simple and effective portable tools that can be used for fast mapping of helminthic diseases and monitoring control programs. Most POC tests (POCTs) available for schistosomiasis diagnosis are lateral flow immunoassays (LFIAs). The emergence of simple and rapid DNA isolation methods, along with isothermal nucleic acid amplification strategies - for example, loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) - and recent clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostic methods facilitate the development of molecular-based POC diagnostics for schistosomiasis. Furthermore, smartphone-based techniques increase real-time connectivity and readout accuracy of POCTs. This review discusses the recent advances in immunological-, molecular-based POCTs and mobile phone microscopes for the diagnosis/screening of schistosomiasis.


Controlled synthesis of hollow octahedral ZnCo2O4 nanocages assembled from ultrathin 2D nanosheets for enhanced lithium storage.
Hollow octahedral ZnCo2O4 nanocages assembled from ultrathin 2D nanosheets are prepared through facile fast simultaneous coordinating etching and thermal processes. Electrochemical results show that the hollow octahedral ZnCo2O4 nanocage is an outstanding anode material for LIBs with a high reversible discharge capacity of 1025 mA h g-1 at 500 mA g-1 after 200 cycles, and an outstanding rate capability of 525 mA h g-1 at 4 A g-1. Moreover, this simple, low cost and fast process could be useful for the construction of many other hollow advanced materials for supercapacitors, sensors and other novel energy and environmental applications.


Relational wellbeing following traumatic interpersonal events and challenges to core beliefs.
Prior research has found that the degree to which a traumatic event challenges core beliefs is associated with adjustment problems; however, how such experiences impact relational wellbeing has received little attention. The current study examined whether negative posttraumatic cognitions mediated the relation between examination of core beliefs and relational wellbeing in young adults following experiences of interpersonal trauma. A moderated parallel mediation model investigated the relation between core beliefs and relational wellbeing through negative cognitions about the self, the world and others, self-blame, and depressive symptoms, following interpersonal violence (IPV; n = 168) and violent loss (VL; n = 102), with ego-resilience moderating the paths from examination of core beliefs to each mediator and relational wellbeing. For individuals with IPV experiences, greater examination of core beliefs was associated with increased depressive symptoms and negative cognitions about the self and about the world and others, each of which was associated with decreased relational wellbeing. For those reporting VL and lower ego-resilience, only negative cognitions about the self emerged as a mediator. Self-blame did not emerge as a mediator whereas ego-resilience emerged as both promotive and protective. Results suggest distinct pathways to relational difficulties following IPV and VL. Following IPV, it was not the process of examining core beliefs that impacted relational functioning, but rather the conclusions drawn about the self, others, and the world. Findings suggest that targeting specific posttraumatic cognitions may enhance relational functioning and possible interventions for addressing such cognitions and fostering ego-resilience are presented. (PsycINFO Database Record


Incidence of atypical nontraumatic diaphyseal fractures of the femur.
Bisphosphonates reduce the rate of osteoporotic fractures in clinical trials and community practice. "Atypical" nontraumatic fractures of the diaphyseal (subtrochanteric or shaft) part of the femur have been observed in patients taking bisphosphonates. We calculated the incidence of these fractures within a defined population and examined the incidence rates according to duration of bisphosphonate use. We identified all femur fractures from January 1, 2007 until December 31, 2011 in 1,835,116 patients older than 45 years who were enrolled in the Healthy Bones Program at Kaiser Southern California, an integrated health care provider. Potential atypical fractures were identified by diagnostic or procedure codes and adjudicated by examination of radiographs. Bisphosphonate exposure was derived from internal pharmacy records. The results showed that 142 patients had atypical fractures; of these, 128 had bisphosphonate exposure. There was no significant correlation between duration of use (5.5 ± 3.4 years) and age (69.3 ± 8.6 years) or bone density (T-score -2.1 ± 1.0). There were 188,814 patients who had used bisphosphonates. The age-adjusted incidence rates for an atypical fracture were 1.78/100,000/year (95% confidence interval [CI], 1.5-2.0) with exposure from 0.1 to 1.9 years, and increased to 113.1/100,000/year (95% CI, 69.3-156.8) with exposure from 8 to 9.9 years. We conclude that the incidence of atypical fractures of the femur increases with longer duration of bisphosphonate use. The rate is much lower than the expected rate of devastating hip fractures in elderly osteoporotic patients. Patients at risk for osteoporotic fractures should not be discouraged from initiating bisphosphonates, because clinical trials have documented that these medicines can substantially reduce the incidence of typical hip fractures. The increased risk of atypical fractures should be taken into consideration when continuing bisphosphonates beyond 5 years.


Transcriptional activity of the human tissue inhibitor of metalloproteinases 1 (TIMP-1) gene in fibroblasts involves elements in the promoter, exon 1 and intron 1.
The active forms of all of the matrix metalloproteinases (MMPs) are inhibited by a family of specific inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Inhibition represents a major level of control of MMP activity. A detailed knowledge of the mechanisms controlling TIMP gene expression is therefore important. We have isolated a genomic clone of the human TIMP-1 gene. A 3 kbp XbaI fragment has been sequenced; this fragment contains 1718 bp 5' flanking sequences, exon 1, a 929 bp intron 1 and part of exon 2. Computer analysis reveals 10 consensus sequences for Sp1, six for activating protein 1 (AP-1), six for polyoma enhancer A3 (PEA3), 12 for AP-2 and five CCAAT boxes. The region hybridizing with a murine TIMP-1 promoter fragment has been subcloned and analysed further. RNase protection identifies six transcription start points, making exon 1 up to 48 bp in length. Transient transfection of promoter-chloramphenicol O-acetyltransferase reporter constructs into primary human connective tissue fibroblasts shows that a 904 bp fragment that hybridizes to a murine TIMP-1 promoter fragment contains a functional promoter. Constructs of -738/+95 to -194/+21 are inducible with serum or phorbol ester to a similar extent to the endogenous TIMP-1 gene. These results and further mapping with 5' deletion mutants from the -738/+95 region have demonstrated that an AP-1 site at -92/-86 is essential for basal expression of the gene. Point mutations within this region have further confirmed the role of this site, along with a more minor role for a neighbouring PEA3 site, in basal expression. Deletions from the 3' end also implicate a region across the exon 1/intron 1 boundary and especially +21 to +58 in basal expression. The +21/+58 region contains a putative binding site for the transcription factor leader-binding protein 1 (LBP-1). Gel-shift analysis shows that protein binds specifically to this region, but competition studies suggest that it is unlikely to be LBP-1.


A Meloidogyne incognita effector is imported into the nucleus and exhibits transcriptional activation activity in planta.
Root-knot nematodes are sedentary biotrophic endoparasites that maintain a complex interaction with their host plants. Nematode effector proteins are synthesized in the oesophageal glands of nematodes and secreted into plant tissue through a needle-like stylet. Effectors characterized to date have been shown to mediate processes essential for nematode pathogenesis. To gain an insight into their site of action and putative function, the subcellular localization of 13 previously isolated Meloidogyne incognita effectors was determined. Translational fusions were created between effectors and EGFP-GUS (enhanced green fluorescent protein-β-glucuronidase) reporter genes, which were transiently expressed in tobacco leaf cells. The majority of effectors localized to the cytoplasm, with one effector, 7H08, imported into the nuclei of plant cells. Deletion analysis revealed that the nuclear localization of 7H08 was mediated by two novel independent nuclear localization domains. As a result of the nuclear localization of the effector, 7H08 was tested for the ability to activate gene transcription. 7H08 was found to activate the expression of reporter genes in both yeast and plant systems. This is the first report of a plant-parasitic nematode effector with transcriptional activation activity.


Iliocaval and Iliofemoral Venous Stenting for Obstruction Secondary to Tumor Compression: Single Center Experience.
Background Cancer patients with pelviabdominal masses can suffer from lower extremity symptoms due to venous compression. The effectiveness of venous stenting has been established in extrinsic venous compression in benign conditions like May-Thurner syndrome. In this retrospective study we evaluate the efficacy and safety of caval, iliocaval and iliofemoral venous stenting for cases of extrinsic venous compression caused by malignant masses in cancer patients. Methods IRB-approved retrospective review of patients who underwent iliofemoral venography with venoplasty and stenting between January 2018 and February 2022 was performed. Patients with extrinsic venous compression caused by malignant masses were included. Data on patient demographics, pre-procedure symptoms, procedural technique, stent characteristics, outcomes and follow-up were collected. Descriptive statistics were used to assess technical success, clinical success, primary stent patency and adverse events of the procedure. Results Thirty-seven patients (19 males, 18 females) who underwent 45 procedures were included. Deep venous thrombosis (DVT) was present in 21 (57%) patients. Twenty-nine patients (78%, 95% CI 62-90%) reported clinical improvement of the presenting symptoms. The median overall survival after the procedure was 4.7 months (95% CI 3.58-5.99). Eight (22%) patients were alive at last follow up with median follow up of 10.33 months (Range 2-25 months). Twenty-six patients had patent stents on their last follow up imaging (70%, 95% CI 61%-91%). Two patients had a small access site hematoma which resolved spontaneously. Two patients developed moderate, and 1 patient developed severe adverse events related to post procedure therapeutic anticoagulation Conclusion Venous stenting is a safe procedure and should be considered as part of the palliative care for patients with debilitating lower extremity symptoms related to iliocaval and iliofemoral venous compression.


Lysine-329 of murine leukemia virus reverse transcriptase: possible involvement in the template-primer binding function.
Treatment of murine leukemia virus reverse transcriptase (MuLV RT) with 4-(oxoacetyl)-phenoxyacetic acid (OAPA) results in the loss of DNA polymerase as well as template-primer binding activity but has no effect on the RT-associated RNase-H activity. Binding stoichiometry revealed that approximately 3 mol of OAPA bound per mole of enzyme, when complete enzyme activation occurred. However, in the presence of template-primer, OAPA does not abolish polymerase activity and 2 mol of OAPA remains bound to 1 mol of enzyme. This observation suggests that only one OAPA reactive site is responsible for the loss of polymerase activity. This site was located on a single tryptic peptide by comparing the maps of the native enzyme and the enzyme treated with OAPA in the presence and absence of template-primer. The appearance of a new peptide peak eluting at 125 min from a C-18 reverse-phase column was consistently noted in the tryptic digest of enzyme treated with OAPA. This peak was absent in tryptic peptides made from the control enzyme or the enzyme protein that was treated with OAPA in the presence of activated DNA or synthetic template-primers. Amino acid composition and sequence analyses of this peptide revealed that it spanned residues 312-342 in the primary amino acid sequence of MuLV RT. Since this peptide does not contain arginine residues and Lys-329 exhibited resistance to tryptic digestion, we conclude that Lys-329 is the target of OAPA action.(ABSTRACT TRUNCATED AT 250 WORDS)


The Dosage Recommendation of Cyclosporin in Children with Hemophagocytic Lymphohistiocytosis Based on Population Pharmacokinetic Model.
Cyclosporin is one of the therapeutic regimens for hemophagocytic lymphohistiocytosis (HLH); however, the optimal dosage of cyclosporine in children with HLH is unknown. It has been found that piperacillin-tazobactam affects the cyclosporine pharmacokinetic process in pediatric HLH patients. Thus, the purpose of the present study was to recommend cyclosporin dosage for pediatric HLH with and without piperacillin- tazobactam. A previously established cyclosporine population pharmacokinetic model for pediatric HLH patients has been used in this study to recommend optimal dosage based on Monte Carlo simulation. The pediatric HLH patients have been included in eight weight groups (5, 10, 20, 30, 40, 50, 60, 70 kg) for sixteen dosages (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 mg/kg), split into one dose or two doses. The optimal cyclosporin dosages for children having HLH without piperacillin-tazobactam have been found to be 15, 13, 12, 11, 10, and 9 mg/kg, split into two doses for weights of 5-7, 7-10, 10-20, 20-28, 28-45, and 45-70 kg, respectively. For children with HLH, optimal cyclosporin dosages with piperacillin-tazobactam have been found to be 8 and 7 mg/kg, split into two doses for weights of 5-20 and 20-70 kg, respectively. It is the first time that the cyclosporin dosage regimens for HLH in children have been developed based on Monte Carlo simulation, and the initial dosage optimizations of cyclosporine in pediatric HLH patients have been recommended.


Annona muricata (Annonaceae): A Review of Its Traditional Uses, Isolated Acetogenins and Biological Activities.
Annona muricata is a member of the Annonaceae family and is a fruit tree with a long history of traditional use. A. muricata, also known as soursop, graviola and guanabana, is an evergreen plant that is mostly distributed in tropical and subtropical regions of the world. The fruits of A. muricata are extensively used to prepare syrups, candies, beverages, ice creams and shakes. A wide array of ethnomedicinal activities is contributed to different parts of A. muricata, and indigenous communities in Africa and South America extensively use this plant in their folk medicine. Numerous investigations have substantiated these activities, including anticancer, anticonvulsant, anti-arthritic, antiparasitic, antimalarial, hepatoprotective and antidiabetic activities. Phytochemical studies reveal that annonaceous acetogenins are the major constituents of A. muricata. More than 100 annonaceous acetogenins have been isolated from leaves, barks, seeds, roots and fruits of A. muricata. In view of the immense studies on A. muricata, this review strives to unite available information regarding its phytochemistry, traditional uses and biological activities.


Predictors of functional outcomes after simple decompression for ulnar neuropathy at the elbow: a multicenter study by the SUN study group.
To identify predictors of surgical outcome for ulnar neuropathy at the elbow (UNE). Prospective cohort followed for 1 year. Clinics. Patients diagnosed with UNE (N=55). All subjects had simple decompression surgery. The primary outcome measure was patient-reported outcomes, such as overall hand function through the Michigan Hand Outcomes Questionnaire (MHQ). Predictors included age, duration of symptoms, disease severity, and motor conduction velocity across the elbow. Multiple regression models with change in the overall MHQ score as the dependent variable showed that at 3 months postoperative time, patients with <3 months duration of symptoms showed 12 points (95% confidence interval [CI], 0.9-23.5) greater improvement in MHQ scores than those with ≥3 months symptom duration. Less than 3 months of symptoms was again associated with 13 points (95% CI, 2.9-24) greater improvement in MHQ scores at 6 months postoperative, but it was no longer associated with better outcomes at 12 months. A worse baseline MHQ score was associated with significant improvement in MHQ scores at 3 months (coefficient, -0.38; 95% CI, -.67 to -.09), and baseline MHQ score was the only significant predictor of 12 month MHQ scores (coefficient, -.40; 95% CI, -.79 to -.01). Subjects with <3 months of symptoms and worse baseline MHQ scores showed significantly greater improvement in functional outcomes as reported by the MHQ. However, duration of symptoms was only predictive at 3 or 6 months because most patients recovered within 3 to 6 months after surgery.


A sodium bicarbonate-acid powered blow-gun syringe for remote injection of wildlife.
An automatic blow-gun syringe which uses carbon dioxide gas as the injecting force is described. Upon striking the animal, carbon dioxide gas is released by the chemical combination of sodium bicarbonate (baking soda) and acid (vinegar), within the blow-gun syringe. The syringe has been used successfully with captive collared peccaries (Dicotyles tajacu). It has the advantages of longer stability, dependable gas expansion, reduction of drug loss, and consistent drug injection.


Nasal pillows as an alternative interface in patients with obstructive sleep apnoea syndrome initiating continuous positive airway pressure therapy.
Side-effects directly due to the nasal mask are common in patients with obstructive sleep apnoea syndrome (OSAS) commencing continuous positive airway pressure (CPAP). Recently, nasal pillows have been designed to overcome these issues. Limited evidence exists of the benefits and effectiveness of these devices. Twenty-one patients (19 male, 49±10years) with the established diagnosis of OSAS [apnoea/hypopnoea index (AHI): 52±22] and who had a successful CPAP titration were commenced on CPAP therapy (10±2cmH2O), and randomized to 4weeks of a nasal pillow (P) and a standard nasal mask (M) in a crossover design. Outcome measures were objective compliance, AHI, quality of life, Epworth Sleepiness Score (ESS) and CPAP side-effects. There was no difference in compliance (M versus P: 5.1±1.9h versus 5.0±1.7h; P=0.701) and AHI (2.6±2.7 versus 3.0±2.9; P=0.509). Quality of life and ESS improved with CPAP, but there was no difference in the extent of improvement between both devices. Usage of nasal pillows resulted in less reported pressure on the face and more subjects found the nasal pillow the more comfortable device. However, there was no clear overall preference for either device at the end of the study (mask=57%, pillow=43%; P=0.513). The applied CPAP pressure did not correlate with compliance, AHI and ESS. Furthermore, no differences in outcome parameters were noted comparing groups with CPAP pressure <10 and ≥10cm H(2) O. Nasal pillows are equally effective in CPAP therapy, but do not generally lead to improved compliance.


Drop-port study of microresonator frequency combs: power transfer, spectra and time-domain characterization.
We use a drop-port geometry to characterize frequency combs generated from silicon nitride on-chip microresonators in the normal group velocity regime. In sharp contrast with the traditional transmission geometry, we observe smooth output spectra with comparable powers in the pump and adjacent comb lines. The power transfer into the comb may be explained to a large extent by the coupling parameters characterizing the linear operation of the resonances studied. Furthermore, comparison of thru- and drop-port spectra shows that much of the ASE noise is filtered out by transmission to the drop-port. Autocorrelation measurements are performed on the drop-port output, without the need to filter out or suppress the strong pump line as is necessary in thru-port experiments. Passively mode-locked pulses with low background are observed in a normal dispersion microcavity.


Regulatory control of the terminal complement proteins at the surface of human endothelial cells: neutralization of a C5b-9 inhibitor by antibody to CD59.
Functionally inhibitory antibody to the plasma membrane complement inhibitor CD59 has been used to investigate control of the terminal complement proteins at the endothelial cell surface. Antibodies against purified human erythrocyte CD59 (polyclonal anti-CD59 and monoclonal antibodies [MoAbs] 1F1 and 1F5) were found to bind specifically to monolayers of cultured human umbilical vein endothelial cells, and by Western blotting to recognize an 18- to 21-Kd endothelial protein. When bound to the endothelial monolayer, anti-CD59 (immunoglobulin G or Fab fragment) potentiated membrane pore formation induced upon C9 binding to C5b-8, and augmented the C5b-9-induced cellular responses, including stimulated secretion of von Willebrand factor and expression of catalytic surface for the prothrombinase enzyme complex. Although potentiating endothelial responses to the terminal complement proteins, anti-CD59 had no effect on the response of these cells to stimulation by histamine. Taken together, these data suggest that human endothelial cells express the CD59 cell surface inhibitor of the terminal complement proteins, which serves to protect these cells from pore-forming and cell-stimulatory effects of the C5b-9 complex. These data also suggest that the inactivation or deletion of this cell surface regulatory molecule would increase the likelihood for procoagulant changes in endothelium exposed to complement activation in plasma.


An experimental investigation of light emission produced in the process of positronium formation in matter.
The excess energy emitted during the positronium (Ps) formation in condensed matter may be released as light. Spectroscopic analysis of this light can be a new method of studying the electronic properties of materials. We report the first experimental attempt, according to our knowledge, to verify the existence of this emission process. As a result, the possibility of the emission of photons during Ps formation is within the experimental uncertainty in two different solids: an n-alkane and porous silica. However, it seems that the Ps formation on the alkane surface is not accompanied by the emission of photons with energy in the detection range of 1.6-3.8 eV. Various processes that can influence the energy of the photon emitted during the Ps formation are discussed to elucidate this issue. To aid future experiments, equations were developed to estimate the expected ratio of light emission events to annihilation events with the presence or absence of a photon during the Ps formation.


A trade-off between reproduction and a condition-dependent sexually selected ornament in the house sparrow Passer domesticus.
Reproductive effort was manipulated in a free-living population of house sparrows (Passer domesticus) to investigate the trade-off between reproductive investment and the expression of a condition-dependent sexually selected ornament. Phenotypic plasticity in the expression of this trait was related to the experimentally manipulated size of the brood reared by a male. Males that invested more in current reproduction subsequently became more attractive to females in this population as they showed a preference for males with smaller badges. This supports the argument that direct benefits are a primary focus for mate choice by females. Trade-offs between reproductive effort and the expression of sexual ornaments are a potentially important source of phenotypic variation in both sexual ornaments and life-history traits.


Understanding and classifying metabolite space and metabolite-likeness.
While the entirety of 'Chemical Space' is huge (and assumed to contain between 10(63) and 10(200) 'small molecules'), distinct subsets of this space can nonetheless be defined according to certain structural parameters. An example of such a subspace is the chemical space spanned by endogenous metabolites, defined as 'naturally occurring' products of an organisms' metabolism. In order to understand this part of chemical space in more detail, we analyzed the chemical space populated by human metabolites in two ways. Firstly, in order to understand metabolite space better, we performed Principal Component Analysis (PCA), hierarchical clustering and scaffold analysis of metabolites and non-metabolites in order to analyze which chemical features are characteristic for both classes of compounds. Here we found that heteroatom (both oxygen and nitrogen) content, as well as the presence of particular ring systems was able to distinguish both groups of compounds. Secondly, we established which molecular descriptors and classifiers are capable of distinguishing metabolites from non-metabolites, by assigning a 'metabolite-likeness' score. It was found that the combination of MDL Public Keys and Random Forest exhibited best overall classification performance with an AUC value of 99.13%, a specificity of 99.84% and a selectivity of 88.79%. This performance is slightly better than previous classifiers; and interestingly we found that drugs occupy two distinct areas of metabolite-likeness, the one being more 'synthetic' and the other being more 'metabolite-like'. Also, on a truly prospective dataset of 457 compounds, 95.84% correct classification was achieved. Overall, we are confident that we contributed to the tasks of classifying metabolites, as well as to understanding metabolite chemical space better. This knowledge can now be used in the development of new drugs that need to resemble metabolites, and in our work particularly for assessing the metabolite-likeness of candidate molecules during metabolite identification in the metabolomics field.


Motion and deformation tracking for short-axis echo-planar myocardial perfusion imaging.
The assessment of regional myocardial perfusion during the first-pass of a contrast agent bolus requires tracking of the signal time course for each myocardial segment so that a detailed perfusion map can be derived. To obtain such a map in practice, however, is not trivial because deformation of the shape of the myocardium and respiratory-induced motion render a major difficulty in this process. This study describes an automated approach for motion and deformation tracking of functional myocardial perfusion images. The effectiveness of the described method has been evaluated using a numerical phantom and results are compared with those from existing techniques which use deformable models. Preliminary results from applying our approach to 20 patients are discussed and compared with those from SPECT studies.


INTERRELATIONSHIPS OF SMOOTH AND NONSMOOTH VARIANTS IN THE DISSOCIATION OF PASTEURELLA HAEMOLYTICA.
Wessman, G. E. (National Animal Disease Laboratory, U.S. Department of Agriculture, Agricultural Research Service, Ames, Iowa). Interrelationships of smooth and nonsmooth variants in the dissociation of Pasteurella haemolytica. J. Bacteriol. 88:356-360. 1964.-Smooth Pasteurella haemolytica, after rapid growth in statically incubated broth cultures, decreased in numbers rapidly, and were replaced by nonsmooth variants. Upon continued incubation, smooth cells again predominated. The two phenotypes were alike in general biochemical characteristics, but differed in virulence for mice. The presence of nonsmooth cells in mixed cultures severely limited the growth of smooth cells. The inhibition of smooth cells correlated with the establishment of definite population densities, and the critical factor appeared to be limitation of oxygen in the cultural medium. Selective inhibition did not occur in aerated cultures, but was more pronounced in cultures grown under reduced air pressure. Selective death of smooth cells on slant cultures held at 5 C and preferential growth of nonsmooth cells, plus death of smooth cells at room temperature, accounted for population changes in stored cultures.


Recurrence-free survival in prostate cancer is related to increased stromal TRAIL expression.
TRAIL (tumor necrosis factor related apoptosis-inducing ligand) is involved in tumor immune surveillance and, thus, may be a potential cancer therapy. TRAIL expression in the tumor microenvironment has been shown to impact cancer survival in multiple tumor types, including ovarian cancer. We studied TRAIL expression and outcomes in patients with prostate cancer. A tissue microarray (TMA) of 200 prostate cancer patients and benign prostate tissue controls was used to assess the epithelial and stromal protein expression of TRAIL, death receptors (DR4 and DR5), decoy receptors (DcR1 and DcR2), and the FLICE inhibitory protein (FLIP(L) ). We correlated these expression patterns with clinicopathological parameters and determined its impact on recurrence-free survival. Nearly all (99.5%) prostate cancer tissues examined displayed either decreased expression of pro-apoptotic TRAIL receptors, increased FLIP(L) expression, or both. We observed elevated death receptor, decoy receptor, FLIP(L) , and epithelial TRAIL expression in prostate cancer epithelium. TRAIL expression in the stromal tumor microenvironment surrounding the prostate cancer was markedly lower. Elevated TRAIL expression in the tumor microenvironment was also significantly associated with increased recurrence-free survival (P = .014), after controlling for other prognostic markers. In contrast, epithelial expression of TRAIL did not have an effect on overall survival. Expression of the components of the pro-apoptotic TRAIL pathway is altered in prostate cancer. Moreover, TRAIL expression in the tumor microenvironment may affect recurrence-free survival rate of prostate cancer patients. Consequently, these results may be useful in devising future therapeutic strategies targeting the TRAIL pathway in prostate cancer.


DDIT3 Immunohistochemistry Is a Useful Tool for the Diagnosis of Myxoid Liposarcoma.
Myxoid liposarcoma is a malignant adipogenic neoplasm characterized by prominent arborizing capillaries, occasional lipoblasts, and primitive-appearing spindle cells in a myxoid background. A recurrent translocation in myxoid liposarcoma results in an oncoprotein consisting of full-length DDIT3 (CHOP) fused to an N-terminal segment of either FUS (TLS) or, less often, EWSR1. Here, we explore the diagnostic significance of DDIT3 expression in myxoid liposarcoma using a mouse monoclonal antibody recognizing an epitope in the N-terminal region. Studying a total of 300 tumors, we find diffuse, moderate-to-strong nuclear-localized anti-DDIT3 immunoreactivity in all 46 cases of myxoid liposarcoma representing 36 unique tumors, including 6 cases with high-grade (round cell) morphology. DDIT3 immunohistochemistry also highlighted a distinctive vasculocentric growth pattern in 7 myxoid liposarcomas treated with neoadjuvant radiation. In contrast, the vast majority of other examined lipomatous and myxoid neoplasms exhibited no DDIT3 expression; limited, weak immunoreactivity in <10% of cells was infrequently observed in dedifferentiated liposarcoma (6/39, 15%), solitary fibrous tumor (3/12, 25%), pleomorphic liposarcoma (1/15, 7%), and high-grade myxofibrosarcoma (2/17, 12%). Although this minimal DDIT3 expression did not correlate with DDIT3 amplification or myxoid liposarcoma-like morphology in dedifferentiated liposarcoma, there was evidence among sarcomas (excluding myxoid liposarcoma) of a relationship between expression and exposure to neoadjuvant radiation or cytotoxic chemotherapy. The constellation of findings indicates that DDIT3 immunohistochemistry may have utility in the evaluation of myxoid and lipomatous neoplasms to support the diagnosis of myxoid liposarcoma.


Bond strength of etched-base-metal alloy to enamel using different etching techniques and cements.
A comparison of bond strength produced by using three different metal surface treatment were evaluated using three different cements. The surface metal treatment were sandblast, electrochemical etching and chemical etching. The three cements used were ABC, Rely-a-bond and panavia EX cements. The following conclusions were obtained: 1. The tensile bond strength of enamel-resin metal joint with panavia EX resin has the best bond strength when the metal was chemically etched. 2. The debonding strength of the chemically etched joints were superior compared to that obtained with either sandblast or electrochemical etching.


Electronic health records for biological sample collection: feasibility study of statin-induced myopathy using the Clinical Practice Research Datalink.
Electronic healthcare records (EHRs) are increasingly used to store clinical information. A secondary benefit of EHRs is their use, in an anonymized form, for observational research. The Clinical Practice Research Datalink (CPRD) contains EHRs from primary care in the UK and, despite 1083 peer-reviewed research publications, has never been used to obtain pharmacogenetic samples. Using a statin-induced myopathy paradigm, we evaluated using the CPRD to obtain patient samples for a pharmacogenetic study targeting 250 cases and 500 controls from UK general practitioner (GP) practices. The CPRD identified potential patients fitting specific case-definition criteria (active rhabdomyolysis or creatine phosphokinase > four times the upper limit of normal), and corresponding GP practices were asked to invite patient participation. Consenting patients were requested to provide either saliva or blood samples and to complete an ethnicity questionnaire. Control subjects were recruited from the same GP practice (saliva) or a small number of practices (blood). Samples were forwarded for DNA extraction. Thirty-six months of recruitment yielded DNA samples from 149 statin-induced myopathy cases and 587 tolerant controls. Data show that contacting patients through their GP is a reliable method for obtaining samples without compromising anonymity. Saliva collection directly from patients was considerably less effective than blood sampling. After 10 months of recruitment, saliva sampling was suspended in favour of blood sampling. We demonstrate the potential of EHRs for identifying accurately phenotyped cases and controls for pharmacogenetic studies. Recruitment was successful only because of the willingness of GP practices to participate and the existence of strong doctor-patient relationships. The present study provides a model that can be implemented in future genetic analyses using EHRs.


Tractional force generation by porcine Müller cells. Development and differential stimulation by growth factors.
To assess the ability of retinal Müller cells to generate tractional forces during dedifferentiation in culture and to assess their responsiveness to contraction-stimulating growth factors. Müller cells were isolated from papain-DNase-digested porcine retina. The identity of the isolated cells was confirmed by immunodetection of carbonic anhydrase II (CA-II), cellular retinaldehyde-binding protein (CRALBP), glial fibrillary acidic protein (GFAP), vimentin, and alpha smooth muscle actin (alpha SMA). Tractional force generation was assessed as a function of Müller cell contraction of collagenous extracellular matrices in vitro. The effects of potential promoters were assessed by addition directly to culture medium. The contributions of specific promoting to the contraction-promoting activity in serum were assessed by adding neutralizing antibodies and measuring loss of stimulatory activity. Freshly isolated Müller cells did not generate substantial matrix contraction. However, this activity increased 150-fold within 12 days in culture and continued to increase during the next 21 days. Development of the capacity for extracellular matrix contraction coincided with the acquisition of immunodetectable alpha SMA and loss of GFAP. Matrix contraction by Müller cells was stimulated in a dose-dependent fashion by human serum, platelet-derived growth factor (PDGF), and insulin-like growth factor-I (IGF-I). Müller cells were not stimulated by transforming growth factor beta 1 (TGF beta 1), transforming growth factor beta 2 (TGF beta 2), or endothelin-1 (E1). Neutralizing antibodies against PDGF and IGF-I reduced the activity in human serum by 37% and 58%, respectively, and 87% when added together. Porcine Müller cells in culture acquire the ability to contract extracellular matrices and thus generate tractional forces. Acquisition of this activity coincides with alpha SMA expression and loss of GFAP. Further, this activity is dependent on the presence of exogenous promoters, including PDGF or IGF-I.


Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR/Cas9.
We have applied the CRISPR/Cas9 system to Drosophila S2 cells to generate targeted genetic mutations in more than 85% of alleles. By targeting a constitutive exon of the AGO1 gene, we demonstrate homozygous mutation in up to 82% of cells, thereby allowing the study of genetic knockouts in a Drosophila cell line for the first time. We have shown that homologous gene targeting is possible at 1-4% efficiency using this system, allowing for the construction of defined insertions and deletions. We demonstrate that a 1 kb homology arm length is optimal for integration by homologous gene targeting, and demonstrate its efficacy by tagging the endogenous AGO1 protein. This technology enables controlled genetic manipulation in Drosophila cell lines, and its simplicity offers the opportunity to study cellular phenotypes genome-wide.


Local control in cardiac E-C coupling.
The development of local control theories in cardiac excitation-contraction coupling solved a major problem in the calcium-induced calcium release (CICR) hypothesis. Local control explained how regeneration, inherent in the CICR mechanism, might be limited spatially to enable graded Ca release (and force production). The key lies in the stochastic recruitment of individual calcium release units (couplons or CRUs) where adjacent CRUs are partially uncoupled by the distance between them. In the CRU, individual groups of sarcoplasmic reticulum calcium release channels (RyRs) are very close to the surface membrane where calcium influx, controlled by membrane depolarization, leads to high local Ca levels that enable a high speed response from RyRs that have a very low probability to opening at resting Ca levels. However, calcium diffusion from an activated CRU results in adjacent CRUs being exposed to much lower levels of Ca and probability of activation. This effectively uncouples the CRUs and limits overall regenerative gain to enable stability without compromising sensitivity. Nevertheless, it is still unclear how the CRU terminates its release of calcium on the physiological timescale, and possible mechanisms (and problems) are briefly reviewed. We suggest that modulation in RyR gating may serve to control average SR Ca levels to regulate other metabolic functions of the sarco(endo)plasmic reticulum beyond regulating contractility. This article is part of a special issue entitled "Local Signaling in Myocytes."


Complement factor B polymorphism 32W protects against age-related macular degeneration.
The 32Q (rs641153; A) and 32W (rs12614; T) variants of complement factor B (CFB) cause less efficient complement activation in vitro than the common 32R variant. This is thought to be the reason that the 32Q variant is associated with decreased risk of age-related macular degeneration (AMD). We investigated whether the 32W variant was also associated with decreased risk of AMD. We genotyped 367 cases with neovascular AMD and 251 disease-free controls. Association with the disease phenotype was assessed by logistic regression for polymorphisms of CFB alone and in combination with smoking status and genetic risk markers of complement factor H (CFH) and HtrA serine peptidase 1 (HTRA1). We performed meta-analysis of all previously published reports of 32W allele frequency in AMD cases and controls. The CFB variant 32W was associated with protection against neovascular AMD, compared to the common 32R variant (odds ratio 0.64, p<0.05, in logistic regression with CFB variants; odds ratio 0.53, p<0.05, in logistic regression with CFB variants, CFH haplotypes, HTRA1 rs10490924 genotype, and smoking status). Meta-analysis (n=1,795) including this study and two others of neovascular AMD showed a combined odds ratio of 0.75 (p<0.05) for 32W, compared to 32R. Meta-analysis (n=2,600) of all reported studies of all types of AMD showed a combined odds ratio of 0.79 (p<0.01). Our study shows that the 32W variant of CFB is associated with protection against AMD, in keeping with evidence of its functional effect on the complement system. The protective effect is less strong than that associated with 32Q.


Retinal charge sensitivity and spatial discrimination obtainable by subretinal implants: key lessons learned from isolated chicken retina.
In order to obtain functional parameters relevant to the designing of a subretinal implant, we carried out electrical stimulation experiments with isolated chicken retina. The median threshold for network activation with planar disc electrodes (diameter 10 microm) was 0.5 nC (625 microC cm(-2)) for anodal voltage impulses and 1.6 nC (2 mC cm(-2)) for cathodal impulses. Above threshold, the number of spikes evoked by a single voltage impulse increased up to saturation within a range of injected charge from 0.1 nC to 1 nC for anodal impulses and from 1 nC to 10 nC for cathodal impulses. Using needle electrodes with a tip diameter of 1 microm, we determined the electrical point spread function (EPSF) for subretinal stimulation. It had a half width in the range of 100 microm, which corresponds to a visual angle of 21' and to a visual acuity of 20/417 in the human eye. It is reasonable to conclude that with subretinal implants the minimum separable will be of the same dimension.


Pharmacodynamics of the type II calcimimetic compound cinacalcet HCl.
Calcimimetic compounds, which activate the parathyroid cell Ca(2+) receptor (CaR) and inhibit parathyroid hormone (PTH) secretion, are under experimental study as a treatment for hyperparathyroidism. This report describes the salient pharmacodynamic properties, using several test systems, of a new calcimimetic compound, cinacalcet HCl. Cinacalcet HCl increased the concentration of cytoplasmic Ca(2+) ([Ca(2+)](i)) in human embryonic kidney 293 cells expressing the human parathyroid CaR. Cinacalcet HCl (EC(50) = 51 nM) in the presence of 0.5 mM extracellular Ca(2+) elicited increases in [Ca(2+)](i) in a dose- and calcium-dependent manner. Similarly, in the presence of 0.5 mM extracellular Ca(2+), cinacalcet HCl (IC(50) = 28 nM) produced a concentration-dependent decrease in PTH secretion from cultured bovine parathyroid cells. Using rat medullary thyroid carcinoma 6-23 cells expressing the CaR, cinacalcet HCl (EC(50) = 34 nM) produced a concentration-dependent increase in calcitonin secretion. In vivo studies in rats demonstrated cinacalcet HCl is orally bioavailable and displays approximately linear pharmacokinetics over the dose range of 1 to 36 mg/kg. Furthermore, this compound suppressed serum PTH and blood-ionized Ca(2+) levels and increased serum calcitonin levels in a dose-dependent manner. Cinacalcet was about 30-fold more potent at lowering serum levels of PTH than it was at increasing serum calcitonin levels. The S-enantiomer of cinacalcet (S-AMG 073) was at least 75-fold less active in these assay systems. The present findings provide compelling evidence that cinacalcet HCl is a potent and stereoselective activator of the parathyroid CaR and, as such, might be beneficial in the treatment of hyperparathyroidism.


Sebelipase alfa in children and adults with lysosomal acid lipase deficiency: Final results of the ARISE study.
Children and adults with lysosomal acid lipase deficiency (LAL-D) experience cirrhosis and dyslipidemia from lysosomal accumulation of cholesteryl esters and triglycerides. Sebelipase alfa enzyme replacement therapy is indicated for individuals with LAL-D. We report final results from the phase III randomized ARISE study of sebelipase alfa in children aged ≥4 years and adults with LAL-D. The study included a 20-week, double-blind, placebo-controlled period; a 130-week, open-label, extension period; and a 104-week, open-label, expanded treatment period. In the open-label periods, all patients received intravenous sebelipase alfa every other week. The primary outcome was alanine aminotransferase (ALT) level normalization; aspartate aminotransferase (AST) levels, lipid parameters, liver histology, liver and spleen volume and fat content, and safety were also assessed. Of 66 patients enrolled, 59 completed the study. Median (range) age at randomization was 13 (4.7-59) years. At the last open-label visit, ALT and AST levels had normalized in 47% and 66% of patients, respectively. Patients who switched from placebo to sebelipase alfa experienced sustained improvements in ALT and AST during the open-label periods that mirrored those observed in the sebelipase alfa group during the double-blind period. Median (IQR) percent changes in lipid levels included a 25% (39%, 6.5%) reduction in low-density lipoprotein cholesterol and a 27% (19%, 44%) increase in high-density lipoprotein cholesterol. Most adverse events during the open-label periods were mild to moderate in severity; 13 patients had infusion-associated reactions (serious in 1 patient). Six patients (9%) developed anti-drug antibodies. Early and rapid improvements in markers of liver injury and lipid abnormalities with sebelipase alfa were sustained, with no progression of liver disease, for up to 5 years. NCT01757184; EudraCT Number: 2011-002750-31 LAY SUMMARY: Sebelipase alfa is used to treat lysosomal acid lipase deficiency (LAL-D), a rare, inherited disease of lipid metabolism. We report the final results of the phase III ARISE clinical study, which show that replacement of the defective LAL enzyme with sebelipase alfa for up to 5 years allows adults and children 4 years of age and older to maintain their initial improvements in liver and cholesterol parameters over the long term, without worsening of liver disease.


The structural basis for the intrinsic disorder of the actin filament: the "lateral slipping" model.
Three-dimensional (3-D) helical reconstructions computed from electron micrographs of negatively stained dispersed F-actin filaments invariably revealed two uninterrupted columns of mass forming the "backbone" of the double-helical filament. The contact between neighboring subunits along the thus defined two long-pitch helical strands was spatially conserved and of high mass density, while the intersubunit contact between them was of lower mass density and varied among reconstructions. In contrast, phalloidinstabilized F-actin filaments displayed higher and spatially more conserved mass density between the two long-pitch helical strands, suggesting that this bicyclic hepta-peptide toxin strengthens the intersubunit contact between the two strands. Consistent with this distinct intersubunit bonding pattern, the two long-pitch helical strands of unstabilized filaments were sometimes observed separated from each other over a distance of two to six subunits, suggesting that the intrastrand intersubunit contact is also physically stronger than the interstrand contact. The resolution of the filament reconstructions, extending to 2.5 nm axially and radially, enabled us to reproducibly "cut out" the F-actin subunit which measured 5.5 nm axially by 6.0 nm tangentially by 3.2 nm radially. The subunit is distinctly polar with a massive "base" pointing towards the "barbed" end of the filament, and a slender "tip" defining its "pointed" end (i.e., relative to the "arrowhead" pattern revealed after stoichiometric decoration of the filaments with myosin subfragment 1). Concavities running approximately parallel to the filament axis both on the inner and outer face of the subunit define a distinct cleft separating the subunit into two domains of similar size: an inner domain confined to radii less than or equal to 2.5-nm forms the uninterrupted backbone of the two long-pitch helical strands, and an outer domain placed at radii of 2-5-nm protrudes radially and thus predominantly contributes to the outer part of the massive base. Quantitative evaluation of successive crossover spacings along individual F-actin filaments revealed the deviations from the mean repeat to be compensatory, i.e., short crossovers frequently followed long ones and vice versa. The variable crossover spacings and diameter of the F-actin filament together with the local unraveling of the two long-pitch helical strands are explained in terms of varying amounts of compensatory "lateral slipping" of the two strands past each other roughly perpendicular to the filament axis. This intrinsic disorder of the actin filament may enable the actin moiety to play a more active role in actin-myosin-based force generation than merely act as a rigid passive cable as has hitherto been assumed.


Clinical Outcomes After Endovascular Repair and Open Surgery to Treat Immunoglobulin G4-Related and Nonrelated Inflammatory Abdominal Aortic Aneurysms.
To compare the follow-up results of endovascular aneurysm repair (EVAR) vs open surgery (OS) for inflammatory abdominal aortic aneurysms (IAAAs) with regard to immunoglobulin G4-related diseases (IgG4-RD), which are fibrous inflammatory conditions characterized by elevated serum IgG4 concentrations and numerous infiltrations of IgG4+ plasmacytes. Between January 2005 and December 2015, 91 patients were treated with EVAR (begun in 2008) and 166 patients underwent OS for AAA. Forty of these 257 patients had IAAAs identified by a >2-mm thickness of periaortic fibrosis (PAF). Of these 40, 21 had pathologically confirmed IgG4-RD and/or serum IgG4 concentrations ≥135 mg/dL (classified IgG4+); 8 (mean age 76 years; 8 men) were treated with EVAR and 13 (mean age 71 years; 11 men) underwent OS. Of the 19 IgG4- patients with IAAA, 9 (mean age 71 years; 8 men) had EVAR and 10 (mean age 75 years; 9 men) had OS. The 4 subgroups were compared in terms of symptoms, complications, inflammation markers, PAF, and aneurysm diameter using the latest midterm follow-up data (12-24 months). Preoperative aneurysm diameter, PAF, gender, median age, symptoms, and median follow-up period were similar in all groups. Preoperative serum IgG4 was equal in EVAR and OS IgG4+ groups. Compared with the OS IgG4+ group, EVAR IgG4+ patients more frequently had postoperative IgG4 increase (5/8; p=0.006) and PAF progression (5/8; p=0.027), higher postoperative serum IgG4 levels (median 141 mg/dL; p=0.034), a thicker postoperative PAF (median 5.1 mm; p=0.016), and persistent clinical symptoms (p=0.006). Compared with EVAR IgG4- patients, the EVAR IgG4+ patients showed significantly thicker postoperative PAF (p=0.024) and larger increases in postoperative sac diameter (median +13.1 mm; p=0.030). Postoperative PAF and sac diameter frequently and synchronously became worse in the EVAR IgG4+ subgroup with increased IgG4 during follow-up. The rate of change in IgG4 significantly positively correlated with the rates of change in PAF (R=0.555, p=0.03) and sac diameter (R=0.902, p=0.003). Though sample sizes were rather small, this pilot study suggested that EVAR-treated IgG4+ IAAA patients have a higher risk of persistent symptoms and increases in PAF, sac diameter, and IgG4 levels. Therefore, OS should be preferred for complete recovery. Frequent monitoring of the postoperative serum IgG4 is necessary following EVAR in IgG4+ patients to detect these complications.


Large molecular forms of cholecystokinin circulating in humans.
Molecular heterogeneity between cholecystokinin (CCK) present in humans and that present in the pig has been proposed. We recently demonstrated that CCK-8 exists in humans in form identical to the porcine peptide. The aims of this work were to evaluate the presence in human plasma of CCK forms larger than CCK-8 and to compare them with the well-characterized porcine forms. Antiserum (no. 4899) was raised in a New Zealand white rabbit immunized with porcine CCK-33 that had specificity for the 7 to 21 region of that peptide and that recognized molecules present in human plasma. To characterize these, postprandial human plasma was applied to an immunoaffinity column generated with this antiserum. Adsorbed peptides were eluted, concentrated on an octadecylsilane cartridge, separated by reversed-phase HPLC and gel filtration chromatography, and screened by cross-reacting and specific CCK and gastrin radioimmunoassays and CCK bioassay by quantification of amylase release by rat pancreatic acini. Two peptides were consistently identified that possessed CCK-like but not gastrin-specific immunoreactivity and CCK-like biological activity. These appeared to be similar in size to CCK-33 and intermediate in size between CCK-33 and CCK-8. Though analogous to porcine CCK based on antibody cross-reactivity and biological activity, the human peptides were heterogeneous from the porcine peptides based on differing chromatographic behavior.


[Various aspects of surgical treatment of vasorenal hypertension].
The results of examination and treatment by surgery of 103 patients with vasorenal hypertension are analysed. Retromboses and restenoses of the reconstructed renal arteries and grafts and uncorrected hyperaldosteronism were the most frequent causes of the poor results. Adequate reconstruction of the renal arteries leads to correction of hyperaldosteronism in patients with normal or high activity of plasma renin. Low-renin secondary hyperaldosteronism in patients with vasorenal hypertension is an indication for simultaneous correction.


Phenolic acids inhibit the formation of advanced glycation end products in food simulation systems depending on their reducing powers and structures.
The concentration of advanced glycation end products (AGEs) in foods, which are formed by Maillard reaction, has demonstrated as risk factors associated with many chronic diseases. The AGEs inhibitory activities of five common phenolic acids (protocatechuic acid, dihydroferulic acid, p-coumaric acid, p-hydroxybenzoic acid and salicylic acid) with different chemical properties had been investigated in two food simulation systems (glucose-bovine serum albumin (BSA) and oleic acid-BSA). The results substantiated that the AGEs inhibitory abilities of phenolic acids in the oleic acid BSA system were much better than the glucose-BSA system for their strong reducing powers and structures. Among them, dihydrogenferulic acid showed strong inhibition of AGEs formation in oleic acid-BSA system at 0.01 mg/mL compared to nonsignificant AGEs inhibitory effect in oleic acid-BSA system at 10-fold higher concentration (0.1 mg/mL). This study suggests that edible plants rich in phenolic acids may be used as AGEs inhibitor during high-fat cooking.


Crop and irrigation management strategies for saline-sodic soils and waters aimed at environmentally sustainable agriculture.
Irrigation has long played a key role in feeding the expanding world population and is expected to play a still greater role in the future. As supplies of good-quality irrigation water are expected to decrease in several regions due to increased municipal-industrial-agricultural competition, available freshwater supplies need to be used more efficiently. In addition, reliance on the use and reuse of saline and/or sodic drainage waters, generated by irrigated agriculture, seems inevitable for irrigation. The same applies to salt-affected soils, which occupy more than 20% of the irrigated lands, and warrant attention for efficient, inexpensive and environmentally acceptable management. Technologically and from a management perspective, a couple of strategies have shown the potential to improve crop production under irrigated agriculture while minimizing the adverse environmental impacts. The first strategy, vegetative bioremediation--a plant-assisted reclamation approach--relies on growing appropriate plant species that can tolerate ambient soil salinity and sodicity levels during reclamation of salt-affected soils. A variety of plant species of agricultural significance have been found to be effective in sustainable reclamation of calcareous and moderately sodic and saline-sodic soils. The second strategy fosters dedicating soils to crop production systems where saline and/or sodic waters predominate and their disposal options are limited. Production systems based on salt-tolerant plant species using drainage waters may be sustainable with the potential of transforming such waters from an environmental burden into an economic asset. Such a strategy would encourage the disposal of drainage waters within the irrigated regions where they are generated rather than exporting these waters to other regions via discharge into main irrigation canals, local streams, or rivers. Being economically and environmentally sustainable, these strategies could be the key to future agricultural and economic growth and social wealth in regions where salt-affected soils exist and/or where saline-sodic drainage waters are generated.


Control of beta-cell differentiation by the pancreatic mesenchyme.
The importance of mesenchymal-epithelial interactions for normal development of the pancreas was recognized in the early 1960s, and mesenchymal signals have been shown to control the proliferation of early pancreatic progenitor cells. The mechanisms by which the mesenchyme coordinates cell proliferation and differentiation to produce the normal number of differentiated pancreatic cells are not fully understood. Here, we demonstrate that the mesenchyme positively controls the final number of beta-cells that develop from early pancreatic progenitor cells. In vitro, the number of beta-cells that developed from rat embryonic pancreatic epithelia was larger in cultures with mesenchyme than without mesenchyme. The effect of mesenchyme was not due to an increase in beta-cell proliferation but was due to increased proliferation of early pancreatic duodenal homeobox-1 (PDX1)-positive progenitor cells, as confirmed by bromodeoxyuridine incorporation. Consequently, the window during which early PDX1(+) pancreatic progenitor cells differentiated into endocrine progenitor cells expressing Ngn3 was extended. Fibroblast growth factor 10 mimicked mesenchyme effects on proliferation of early PDX1(+) progenitor cells and induction of Ngn3 expression. Taken together, our results indicate that expansion of early PDX1(+) pancreatic progenitor cells represents a way to increase the final number of beta-cells developing from early embryonic pancreas.