Yeast ribosomal protein L40 assembles late into precursor 60 S ribosomes and is required for their cytoplasmic maturation.
Most ribosomal proteins play important roles in ribosome biogenesis and function. Here, we have examined the contribution of the essential ribosomal protein L40 in these processes in the yeast Saccharomyces cerevisiae. Deletion of either the RPL40A or RPL40B gene and in vivo depletion of L40 impair 60 S ribosomal subunit biogenesis. Polysome profile analyses reveal the accumulation of half-mers and a moderate reduction in free 60 S ribosomal subunits. Pulse-chase, Northern blotting, and primer extension analyses in the L40-depleted strain clearly indicate that L40 is not strictly required for the precursor rRNA (pre-rRNA) processing reactions but contributes to optimal 27 SB pre-rRNA maturation. Moreover, depletion of L40 hinders the nucleo-cytoplasmic export of pre-60 S ribosomal particles. Importantly, all these defects most likely appear as the direct consequence of impaired Nmd3 and Rlp24 release from cytoplasmic pre-60 S ribosomal subunits and their inefficient recycling back into the nucle(ol)us. In agreement, we show that hemagglutinin epitope-tagged L40A assembles in the cytoplasm into almost mature pre-60 S ribosomal particles. Finally, we have identified that the hemagglutinin epitope-tagged L40A confers resistance to sordarin, a translation inhibitor that impairs the function of eukaryotic elongation factor 2, whereas the rpl40a and rpl40b null mutants are hypersensitive to this antibiotic. We conclude that L40 is assembled at a very late stage into pre-60 S ribosomal subunits and that its incorporation into 60 S ribosomal subunits is a prerequisite for subunit joining and may ensure proper functioning of the translocation process.


Isolation of a potent antibiotic producer bacterium, especially against MRSA, from northern region of the Persian Gulf.
Nowadays, emergence and prevalence of MRSA (Methicillin Resistant Staphylococcus aureus) strain have become a great global concern in 21st century, so, it is necessary to discover new antibiotics against this pathogen. The aim of this study was isolation and evaluation marine bacteria from the Persian Gulf in order to finding antibiotic compounds against some pathogenic bacteria. For this purpose, water and sediment samples were collected from the Persian Gulf during March to October 2009. The antibacterial activity of the isolated bacteria was assessed using disc diffusion method. The Growth Curve Interference (GCI) parameter against MRSA was determined for the high potential antibiotic producing strain. The most important factors affecting fermentation conditions in antibiotic production were also optimized. Definite identification of intended isolate was confirmed by 16S rRNA sequencing. Altogether, 51 bacterial colony was isolated and among them only 3 bacterium showed antibacterial activity. Pseudoalteromonas piscicida PG-01 isolated from a sediment sample was chosen as the best antibiotic producing strain. This strain was effective against all tested Gram-positive bacteria, had good anti-MRSA activity and also GCI value against MRSA was two times lower than MIC value. Among the optimized fermentation parameters, carbon and nitrogen sources play major role in efficacy of optimized antibiotic production. Ultrastructural study on the effect of intended antibiotic compounds on MRSA using TEM revealed that the target site for this compound is cell wall. Considering the antibacterial effect of PG-01 strain especially against MRSA, intended antibiotic compounds can gives hope for treatment of diseases caused by multi-drug resistant bacteria.


The zebra finch, Taeniopygia guttata: an avian model for investigating the neurobiological basis of vocal learning.
Songbirds are capable of learning their vocalizations by copying a singing adult. This vocal learning ability requires juveniles to hear and memorize the sound of the adult song, and later to imitate it through a process involving sensorimotor integration. Vocal learning is a trait that songbirds share with humans, where it forms the basis of spoken language acquisition, with other avian groups (parrots and hummingbirds), and with a few other mammals (cetaceans, bats). It is however absent in traditional model organisms such as rodents and nonhuman primates. Zebra finches, a songbird species from Australia, are popular pets and are easy to breed. They also sing a relatively simple and stereotyped song that is amenable to quantitative analysis. Zebra finches have thus emerged as a choice model organism for investigating the neurobiological basis of vocal learning. A number of tools and methodologies have been developed to characterize the bioacoustics properties of their song, analyze the degree of accurate copying during vocal learning, map the brain circuits that control singing and song learning, and investigate the physiology of these circuits. Such studies have led to a large base of knowledge on song production and learning, and their underlying neural substrate. Several molecular resources have recently become available, including brain cDNA/EST databases, microarrays, BAC libraries, a molecular brain atlas, a complete genome assembly, and the ability to perform transgenesis. The recent availability of many other avian genomes provides unique opportunities for comparative analysis in the search for features unique to vocal learning organisms.


Endogenous hydrogen peroxide increases biofilm formation by inducing exopolysaccharide production in Acinetobacter oleivorans DR1.
In this study, we investigated differentially expressed proteins in Acinetobacter oleivorans cells during planktonic and biofilm growth by using 2-dimensional gel electrophoresis combined with matrix-assisted laser desorption time-of-flight mass spectrometry. We focused on the role of oxidative stress resistance during biofilm formation using mutants defective in alkyl hydroperoxide reductase (AhpC) because its production in aged biofilms was enhanced compared to that in planktonic cells. Results obtained using an ahpC promoter-gfp reporter vector showed that aged biofilms expressed higher ahpC levels than planktonic cells at 48 h. However, at 24 h, ahpC expression was higher in planktonic cells than in biofilms. Deletion of ahpC led to a severe growth defect in rich media that was not observed in minimal media and promoted early biofilm formation through increased production of exopolysaccharide (EPS) and EPS gene expression. Increased endogenous H2O2 production in the ahpC mutant in rich media enhanced biofilm formation, and this enhancement was not observed in the presence of antioxidants. Exogenous addition of H2O2 promoted biofilm formation in wild type cells, which suggested that biofilm development is linked to defense against H2O2. Collectively, our data showed that EPS production caused by H2O2 stress enhances biofilm formation in A. oleivorans.


A simple recipe for sophisticated cocktails: organocatalytic one-pot reactions--concept, nomenclature, and future perspectives.
Asymmetric organocatalysis has been successfully incorporated in many multistep one-pot sequences to provide simple access to structurally complex target molecules in a highly stereoselective fashion. The key feature behind this success is the ability of organocatalyzed reactions to proceed efficiently in the presence of large amounts of spectator reagents. Additionally, owing to their organic nature and substoichiometric presence, organocatalysts are also expected to become innocent bystanders in subsequent transformations. In this Minireview, an easy-to-use classification and nomenclatural system that is capable of systematically and informatively describing each one-pot reaction is introduced, and selected important contributions within the field of organocatalytic one-pot reactions are reviewed according to this new system. Finally, future developments and perspectives in the field are discussed.


Au growth on semiconductor nanorods: photoinduced versus thermal growth mechanisms.
Gold growth on CdS nanorods and on seeded CdSe/CdS nanorods with and without illumination at different temperatures was studied. Two competing mechanisms were identified: thermal and light-induced growth. The thermal mechanism leads to growth of small gold particles at defects along the rod body and can be suppressed at lower temperatures. This control is attributed to a phase transition of the alkyl chains of the surface amine ligands to a static phase at lower temperatures, blocking the Au precursor's access to the nanorod surfaces. While a long-chain (C18) amine shows effective blocking at 293 K, a shorter chain (C12) amine shows the same result only at 273 K; however, in the case of a bulky trialkylamine, defect growth was observed even at 273 K. Light-induced growth leads to selective deposition of gold on one end of the rods. The tip was shown to grow on sulfur-rich facets of the nanorod, producing end-on and angled tip orientations. Growth under illumination with decreased temperature provides a highly selective synthesis of hybrid semiconductor nanorods with a single gold tip. Such anisotropic semiconductor-metal hybrids are of interest for self-assembly and photocatalysis and as building blocks in optoelectronic devices.


Association Between Income and Perinatal Mortality in the Netherlands Across Gestational Age.
The association between household income and perinatal health outcomes has been understudied. Examining disparities in perinatal mortality within strata of gestational age and before and after adjusting for birth weight centile can reveal how the income gradient is associated with gestational age, birth weight, and perinatal mortality. To investigate the association between household income and perinatal mortality, separately by gestational age strata and time of death, and the potential role of birth weight centile in mediating this association. This cross-sectional study used individually linked data of all registered births in the Netherlands with household-level income tax records. Singletons born between January 1, 2004, and December 31, 2016, at 24 weeks to 41 weeks 6 days of gestation with complete information on birth outcomes and maternal characteristics were studied. Data analysis was performed from March 1, 2018, to August 30, 2021. Household income rank (adjusted for household size). Perinatal mortality, stillbirth (at ≥24 weeks of gestation), and early neonatal mortality (at ≤7 days after birth). Disparities were expressed as bottom-to-top ratios of projected mortality among newborns with the poorest 1% of households vs those with the richest 1% of households. Generalized additive models stratified by gestational age categories, adjusted for potential confounding by maternal age at birth, maternal ethnicity, parity, sex, and year of birth, were used. Birth weight centile was included as a potential mediator. Among 2 036 431 singletons in this study (1 043 999 [51.3%] males; 1 496 579 [73.5%] with mother of Dutch ethnicity), 121 010 (5.9%) were born before 37 weeks of gestation, and 8720 (4.3 deaths per 1000) died during the perinatal period. Higher household income was positively associated with higher rates of perinatal survival, with an unadjusted bottom-to-top ratio of 2.18 (95% CI, 1.87-2.56). The bottom-to-top ratio decreased to 1.30 (95% CI, 1.22-1.39) after adjustment for potential confounding factors and inclusion of birth weight centile as a possible mediator. The fully adjusted ratios were lower for stillbirths (1.27; 95% CI, 1.20-1.36) than for early neonatal deaths (1.35; 95% CI, 1.14-1.66). Inequalities in perinatal mortality were found for newborns at greater than 26 weeks of gestation but not between 24 and 26 weeks of gestation (fully adjusted bottom-to-top ratio, 0.89; 95% CI, 0.77-1.04). The results of this large nationally representative cross-sectional study suggest that a large part of the increased risk of perinatal mortality among low-income women is mediated via a lower birth weight centile. The absence of disparities at very low gestational ages suggests that income-related risk factors for perinatal mortality are less prominent at very low gestational ages. Further research should aim to understand which factors associated with preterm birth and lower birth weight can reduce inequalities in perinatal mortality.


Multifactorial inheritance with cultural transmission and assortative mating. III. Family structure and the analysis of separation experiments.
Demographic data about family composition or structure in the United States is reviewed. About 25% of white children and a majority of black children are reared in either broken or extended families, and this must be taken into consideration for valid studies of cultural inheritance. Atypical family structures are described including those in which parents include: biological parents, stepparents, grandparents, uncles, aunts, sibs, foster parents, and their spouses. General formulae for a wide variety of kinship correlations are derived using path analysis. The multifactorial model presented allows for cultural inheritance, polygenic inheritance, correlated sibling environments, and phenotypic assortative mating (as previously described for intact families) plus extensions necessary for the analysis of separation experiments. These extensions allow for variable family structure and differences in parental influence due to separation, age or stage of development of the child, birth order, or type of relationship. Family structure is observed to have a marked effect on familial resemblance. Computer simulation studies demonstrate marked heterogeneity among phenotypic correlations for kinships of the same degree of genetic relationship arising in different family structures. Analyses of multiple types of sibs and other relatives in variable family structures offer great promise for the study of cultural inheritance.


Specialist Herbivore Performance on Introduced Plants During Native Host Decline.
Ash (Fraxinus spp.) is in rapid decline across the northeastern USA due to the invasive emerald ash borer (Agrilus planipennis Fairmaire). Three recently co-occurring confamilial species may serve as alternative larval host plants for ash-reliant Lepidoptera. These prospective hosts are nonnative shrubs often planted in managed suburban landscapes and are sometimes invasive or naturalized in North America. Given the imminent decline of ash trees, we considered potential downstream effects on insect herbivores historically specialized on ash foliage. We measured the performance of three ash-specialist hawkmoths (Lepidoptera: Sphingidae) on native white ash (Fraxinus americana L.) and alternative host plants: common lilac (Syringa vulgaris L.), weeping forsythia [Forsythia suspensa (Thunb.) Vahl], and European privet (Ligustrum vulgare L.). We found the nonnative host plants provided varied support for larval survival to pupation, with biomass and growth rate affected differently by both plant and insect identity. Nearly all caterpillars reared on one alternative host, European privet, exhibited distinct malformations of the wing buds at pupation. Given caterpillar presence on privet in the field, privet may constitute an ecological trap (i.e., when female moths select a sub-optimal host, offspring survival and fitness are reduced). This work demonstrates how performance testing can reveal species-specific effects of host plant loss on mono- or oligophagous insects. For some ash specialists, alternative nonnative host plants may be suboptimal, but some cultivated host plants may be able to support certain specialist insects during native host decline. We suggest that landscaping decisions can be tailored to support threatened insect species.


Transformation invariant on-line target recognition.
Transformation invariant automatic target recognition (ATR) has been an active research area due to its widespread applications in defense, robotics, medical imaging and geographic scene analysis. The primary goal for this paper is to obtain an on-line ATR system for targets in presence of image transformations, such as rotation, translation, scale and occlusion as well as resolution changes. We investigate biologically inspired adaptive critic design (ACD) neural network (NN) models for on-line learning of such transformations. We further exploit reinforcement learning (RL) in ACD framework to obtain transformation invariant ATR. We exploit two ACD designs, such as heuristic dynamic programming (HDP) and dual heuristic dynamic programming (DHP) to obtain transformation invariant ATR. We obtain extensive statistical evaluations of proposed on-line ATR networks using both simulated image transformations and real benchmark facial image database, UMIST, with pose variations. Our simulations show promising results for learning transformations in simulated images and authenticating out-of plane rotated face images. Comparing the two on-line ATR designs, HDP outperforms DHP in learning capability and robustness and is more tolerant to noise. The computational time involved in HDP is also less than that of DHP. On the other hand, DHP achieves a 100% success rate more frequently than HDP for individual targets, and the residual critic error in DHP is generally smaller than that of HDP. Mathematical analyses of both our RL-based on-line ATR designs are also obtained to provide a sufficient condition for asymptotic convergence in a statistical average sense.


ON THE PERMEABILITY OF THE STOMACH MUCOSA FOR ACIDS AND SOME OTHER SUBSTANCES.
1. Solutions approximately isotonic with blood of strong and weak acids, several salts, glucose, and glycine were introduced in the resting stomachs of cats. The concentration and volume changes were recorded. 2. It was found that the stomach mucosa was permeable to the majority of the ions tested. There was also a permeability in the opposite direction from the blood (mucosa) to the stomach content, particularly of alkali chlorides. Poorly permeable substances were glucose, glycine, and sodium iodate. Pure weak acids such as acetic acid penetrated very rapidly. 3. The electrolyte concentration changes in the stomach content (or gastric juice) are pictured as an exchange diffusion; for instance, the hydrogen ions of an acid are exchanged against alkali ions of the mucosa or blood. 4. It is pointed out that the concept of the mucosa as an ion permeable membrane could be used as the foundation of a "diffusion theory," which can explain the acidity and chloride variations of the gastric juice without postulating neutralizing or diluting secretions.


Frontiers in human factors: integrating human factors and ergonomics to improve safety and quality in Latin American healthcare systems.
The importance of human factors/ergonomics (HFE) is well established in all high-reliability systems but only applied in the healthcare sector relatively recently. Across many sectors, low-/middle-income countries (LMICs) lag behind more economically developed countries in their application of this safety science, due to resource and, in some cases, awareness and expertise. Most previous applications of HFE related to occupational ergonomics rather than healthcare safety. The paper details how the reputation of HFE is being developed within healthcare communities of Latin America (LatAm), through increasing awareness and understanding of its role as safety science in the healthcare sector. It starts by articulating the need for HFE and then provides examples from Mexico, Colombia and Peru. The practical examples for research and education illustrate a developing awareness of the relevance of HFE to the healthcare sectors in LatAm and an appreciation of its worth to improve health service quality and patient safety through healthcare community engagement. A new LatAm Network of HFE in Healthcare Systems (RELAESA) was formed in 2019, which has provided a platform for HFE advice during the COVID-19 pandemic. There is a real opportunity in LatAm and other LMIC health services to make more rapid and sustainable progress in healthcare-embedded HFE than has been experienced within healthcare services of more developed nations.


The subventricular zone is able to respond to a demyelinating lesion after localized radiation.
Radiation is a common tool in the treatment of brain tumors that induces neurological deficits as a side effect. Some of these deficits appear to be related to the impact of radiation on the neurogenic niches, producing a drastic decrease in the proliferative capacity of these regions. In the adult mammalian brain, the subventricular zone (SVZ) of the lateral ventricles is the main neurogenic niche. Neural stem/precursor cells (NSCs) within the SVZ play an important role in brain repair following injuries. However, the irradiated NSCs' ability to respond to damage has not been previously elucidated. In this study, we evaluated the effects of localized radiation on the SVZ ability to respond to a lysolecithin-induced demyelination of the striatum. We demonstrated that the proliferation rate of the irradiated SVZ was increased after brain damage and that residual NSCs were reactivated. The irradiated SVZ had an expansion of doublecortin positive cells that appeared to migrate from the lateral ventricles toward the demyelinated striatum, where newly generated oligodendrocytes were found. In addition, in the absence of demyelinating damage, remaining cells in the irradiated SVZ appeared to repopulate the neurogenic niche a year post-radiation. These findings support the hypothesis that NSCs are radioresistant and can respond to a brain injury, recovering the neurogenic niche. A more complete understanding of the effects that localized radiation has on the SVZ may lead to improvement of the current protocols used in the radiotherapy of cancer.


Morbidity and Mortality of Spinal Epidural Abscess in End-Stage Renal Disease Patients: A Case-Control Study.
Spinal epidural abscess (SEA) is an uncommon and highly morbid infection of the epidural space. End-stage renal disease (ESRD) patients are known to be at increased risk of developing SEA; however, there are no studies that have described the risk factors and outcomes of SEA in ESRD patients utilizing the United States Renal Data System (USRDS). To determine risk factors, morbidity, and mortality associated with SEA in ESRD patients, a retrospective case-control study was conducted using the USRDS. ESRD patients diagnosed with SEA between 2005 and 2010 were identified, and logistic regression was performed to examine correlates of SEA, as well as risk factors associated with mortality in SEA-ESRD patients. The prevalence of SEA amongst ESRD patients was 0.39% (n = 1,697). Patients with SEA were more likely to be male [adjusted Odds Ratio (OR) = 1.22], black (OR = 1.19), diabetic (OR = 1.26), with catheter access (OR = 1.29), and less likely to be ≥65 years old (OR = 0.64). Osteomyelitis, bacteremia/septicemia, MRSA, and endocarditis were all significantly associated with increased risk of SEA (OR = 1.54-5.14). Age ≥65 years (HR = 1.45), urinary tract infections (HR = 1.26), decubitus ulcers (HR=1.37), and post-SEA paraplegia (HR = 1.25) were significantly associated with mortality among those with SEA. As described in previous literature, risk factors for SEA included infections, diabetes, and indwelling catheters. Additionally, clinicians should be aware of the risk factors for mortality in SEA-ESRD patients. As the largest study of SEA to date, our report identifies important risk factors for SEA in ESRD patients, and novel data regarding their mortality-associated risk factors.


Comparing current definitions of return to work: a measurement approach.
Return-to-work (RTW) status is an often used outcome in work and health research. In low back pain, work is regarded as a normal activity a worker should return to in order to fully recover. Comparing outcomes across studies and even jurisdictions using different definitions of RTW can be challenging for readers in general and when performing a systematic review in particular. In this study, the measurement properties of previously defined RTW outcomes were examined with data from two studies from two countries. Data on RTW in low back pain (LBP) from the Canadian Early Claimant Cohort (ECC); a workers' compensation based study, and the Dutch Amsterdam Sherbrooke Evaluation (ASE) study were analyzed. Correlations between outcomes, differences in predictive validity when using different outcomes and construct validity when comparing outcomes to a functional status outcome were analyzed. In the ECC all definitions were highly correlated and performed similarly in predictive validity. When compared to functional status, RTW definitions in the ECC study performed fair to good on all time points. In the ASE study all definitions were highly correlated and performed similarly in predictive validity. The RTW definitions, however, failed to compare or compared poorly with functional status. Only one definition compared fairly on one time point. Differently defined outcomes are highly correlated, give similar results in prediction, but seem to differ in construct validity when compared to functional status depending on societal context or possibly birth cohort. Comparison of studies using different RTW definitions appears valid as long as RTW status is not considered as a measure of functional status.


Physician, heal thyself: health maintenance behaviors among physicians.
The goal of this article is to assess the need for health maintenance intervention programs directed at physicians and patients. We compared the health maintenance behavior compliance of physicians (as patients) to a control patient population. An online survey was sent to the Texas Tech University Health Sciences Center (TTUHSC) medical school clinical and nonclinical basic science faculties and to the non-TTUHSC clinicians of the local county medical society. The survey included questions regarding basic demographics and recent participation in regular health maintenance strategies including annual checkup, influenza vaccination, colonoscopy, Pap smear, and mammogram. Logistic regressions were conducted to determine if being a physician had a significant association with the likelihood of participating in the health maintenance behavior outcomes listed above. This article shows that physicians are less likely than the general population to adhere to specific health maintenance guidelines: namely, annual checkups, colonoscopies, and mammograms. Pap smear rates did not differ between physicians and a control population, but physicians showed an increased likelihood of receiving an influenza vaccine.


Effect of warm or cold exposure on GDP binding and uncoupling protein in rat brown fat.
The effects of acute and chronic exposure to different environmental temperatures on the total tissue cytochrome oxidase activity, level of mitochondrial GDP binding, and specific mitochondrial concentration of uncoupling protein have been investigated in rat brown adipose tissue, a radioimmunoassay being used to measure uncoupling protein. Acclimation at different temperatures for 3 wk produced parallel changes in GDP binding, the concentration of uncoupling protein, and the activity of cytochrome oxidase, each parameter rising with decreasing temperature between thermoneutrality (29 degrees C) and 4 degrees C. Acute exposure of warm-acclimated (29 degrees C) rats to the cold (4 degrees C) led to a rapid increase in GDP binding without any alteration in the amount of uncoupling protein. The increase in binding was accompanied by an increase in the rate of acetate-induced swelling of the mitochondria. The concentration of uncoupling protein in warm-acclimated rats was significantly raised only after 48 h exposure to cold. When cold-acclimated rats were exposed acutely to the warm, there was a rapid decrease in GDP binding without any alteration in the amount of uncoupling protein. It is concluded that after alterations in environmental temperature the concentration of uncoupling protein in brown adipose tissue mitochondria changes much more slowly than GDP binding and that binding can therefore be dissociated from the amount of the protein.


Effect of cough machine assistance and care bundle combined therapy on burned patients with moderate to severe inhalation injury.
To investigate the effectiveness of cough machine assistance combined with care bundle on mental resilience and self-efficacy on burned patients with moderate to severe inhalation injury.Totally 98 burned patients with moderate to severe inhalation injury from April 2021 to April 2022 were enrolled in this retrospective analysis. They were divided into the control group (n=49, conventional therapy) and combined-therapy group (n=49, cough machine assistance and care bundles). The mental resilience, posttraumatic growth, and self-efficacy were conducted. The arterial blood gas analysis indicators and incidence of pulmonary infection were also evaluated.The length of hospital stay in combined-therapy group was significantly less than that of control group. There was no difference of pulmonary infections between two groups before treatment. However, the incidence of pulmonary infection in the combined-therapy group was significantly lower. After the therapy, the combined-therapy group showed significantly higher partial pressure of oxygen, and lower levels of oxyhemoglobin saturation and partial pressure of carbon dioxide compared with the control group. The evaluation with the mental resilience, posttraumatic growth and self-efficacy after the therapy, indicated that the patients in the combined-therapy group had significantly higher scores of optimism, strength, tenacity, relating to others, new possibilities, personal strength, appreciation of life and spiritual change, and self-efficacy.The combined therapy with cough machine assistance and care bundles can effectively improve the mental resilience and self-efficacy level of burned patients with moderate to severe inhalation injury, showing potential for possible clinical applications.


Avian viral surveillance in Victoria, Australia, and detection of two novel avian herpesviruses.
Viruses in avian hosts can pose threats to avian health and some have zoonotic potential. Hospitals that provide veterinary care for avian patients may serve as a site of exposure of other birds and human staff in the facility to these viruses. They can also provide a useful location to collect samples from avian patients in order to examine the viruses present in wild birds. This study aimed to investigate viruses of biosecurity and/or zoonotic significance in Australian birds by screening samples collected from 409 birds presented to the Australian Wildlife Health Centre at Zoos Victoria's Healesville Sanctuary for veterinary care between December 2014 and December 2015. Samples were tested for avian influenza viruses, herpesviruses, paramyxoviruses and coronaviruses, using genus- or family-wide polymerase chain reaction methods coupled with sequencing and phylogenetic analyses for detection and identification of both known and novel viruses. A very low prevalence of viruses was detected. Columbid alphaherpesvirus 1 was detected from a powerful owl (Ninox strenua) with inclusion body hepatitis, and an avian paramyxovirus most similar to Avian avulavirus 5 was detected from a musk lorikeet (Glossopsitta concinna). Two distinct novel avian alphaherpesviruses were detected in samples from a sulphur-crested cockatoo (Cacatua galerita) and a tawny frogmouth (Podargus strigoides). Avian influenza viruses and avian coronaviruses were not detected. The clinical significance of the newly detected viruses remains undetermined. Further studies are needed to assess the host specificity, epidemiology, pathogenicity and host-pathogen relationships of these novel viruses. Further genome characterization is also indicated, and would be required before these viruses can be formally classified taxonomically. The detection of these viruses contributes to our knowledge on avian virodiversity. The low level of avian virus detection, and the absence of any viruses with zoonotic potential, suggests low risk to biosecurity and human health.


Clearing-factor lipase in adipose tissue. Distinction of different states of the enzyme and the possible role of the fat cell in the maintenance of tissue activity.
1. Incubation of intact epididymal adipose tissue from fed rats at 37 degrees in an albumin solution at pH7.4 in vitro results in rapid loss of clearing-factor lipase activity until a low activity, stable to prolonged incubation, is attained. The clearing-factor lipase activity of intact tissue from starved rats, which is initially much less than that of tissue from fed rats, is mainly stable to incubation at 37 degrees . 2. Much of the clearing-factor lipase activity of intact epididymal adipose tissue from fed rats is inactivated by collagenase. The enzyme activity of intact tissue from starved rats is not inactivated by collagenase. 3. The clearing-factor lipase activity of fat cells isolated from the epididymal adipose tissue of fed rats is stable to prolonged incubation at 37 degrees . It represents only a small proportion of the total activity of the intact tissue. In starved rats, the isolated fat cells contain a much higher proportion of the activity of the intact tissue. Their activity is also stable at 37 degrees . 4. Incubation of isolated fat cells in a serum-based medium leads to a progressive rise in clearing-factor lipase activity. Actinomycin increases the extent of this rise in activity. No rise in clearing-factor lipase activity occurs when stromal-vascular cells isolated from epididymal adipose tissue are incubated in the medium. 5. The findings indicate that less than 20% of the activity of intact adipose tissue from fed rats is retained when fat cells are isolated from the tissue by collagenase treatment. The activity that is lost could be that which normally functions in the uptake of triglyceride fatty acids by the tissue.


Valuing Hemlock Woolly Adelgid Control in Public Forests: Scope Effects with Attribute Non-Attendance.
Sensitivity to the scope of public good provision is an important indication of validity for the contingent valuation method. An online survey was administered to an opt-in non-probability sample panel to estimate the willingness-to-pay to protect hemlock trees from a destructive invasive species on federal land in North Carolina. We collected survey responses from 907 North Carolina residents. We find evidence that attribute non-attendance (ANA) is a factor when testing for sensitivity to scope. When estimating the model with stated ANA, the ecologically and socially important scope coefficients become positive and statistically significant with economically significant marginal willingness-to-pay estimates.


Pediatric Cholestatic Liver Disease: Review of Bile Acid Metabolism and Discussion of Current and Emerging Therapies.
Cholestatic liver diseases are a significant cause of morbidity and mortality and the leading indication for pediatric liver transplant. These include diseases such as biliary atresia, Alagille syndrome, progressive intrahepatic cholestasis entities, ductal plate abnormalities including Caroli syndrome and congenital hepatic fibrosis, primary sclerosing cholangitis, bile acid synthesis defects, and certain metabolic disease. Medical management of these patients typically includes supportive care for complications of chronic cholestasis including malnutrition, pruritus, and portal hypertension. However, there are limited effective interventions to prevent progressive liver damage in these diseases, leaving clinicians to ultimately rely on liver transplantation in many cases. Agents such as ursodeoxycholic acid, bile acid sequestrants, and rifampicin have been mainstays of treatment for years with the understanding that they may decrease or alter the composition of the bile acid pool, though clinical response to these medications is frequently insufficient and their effects on disease progression remain limited. Recently, animal and human studies have identified potential new therapeutic targets which may disrupt the enterohepatic circulation of bile acids, alter the expression of bile acid transporters or decrease the production of bile acids. In this article, we will review bile formation, bile acid signaling, and the relevance for current and newer therapies for pediatric cholestasis. We will also highlight further areas of potential targets for medical intervention for pediatric cholestatic liver diseases.


Algorithms for Automated Calibration of Transcutaneous Spinal Cord Stimulation to Facilitate Clinical Applications.
Transcutaneous spinal cord stimulation (tSCS) is a promising intervention that can benefit spasticity control and augment voluntary movement in spinal cord injury (SCI) and multiple sclerosis. Current applications require expert knowledge and rely on the thorough visual analysis of electromyographic (EMG) responses from lower-limb muscles to optimize attainable treatment effects. Here, we devised an automated tSCS setup by combining an electrode array placed over low-thoracic to mid-lumbar vertebrae, synchronized EMG recordings, and a self-operating stimulation protocol to systematically test various stimulation sites and amplitudes. A built-in calibration procedure classifies the evoked responses as reflexes or direct motor responses and identifies stimulation thresholds as recommendations for tSCS therapy. We tested our setup in 15 individuals (five neurologically intact, five SCI, and five Parkinson's disease) and validated the results against blinded ratings from two clinical experts. Congruent results were obtained in 13 cases for electrode positions and in eight for tSCS amplitudes, with deviations of a maximum of one position and 5 to 10 mA in amplitude in the remaining cases. Despite these minor deviations, the calibration found clinically suitable tSCS settings in 13 individuals. In the remaining two cases, the automatic setup and both experts agreed that no reflex responses could be detected. The presented technological developments may facilitate the dissemination of tSCS into non-academic environments and broaden its use for diagnostic and therapeutic purposes.


Radiosensitization of colon cancer cell lines by docetaxel: mechanisms of action.
The radiation-modifying action of docetaxel in experimental systems is well established. Docetaxel is also an increasingly important drug for the treatment of cancer in concurrent radiotherapy protocols. However, the mechanisms of docetaxel radiosensitization are not fully understood. We have investigated the magnitude and mechanisms of docetaxel radiosensitization in vitro in four human colorectal cancer cell lines (SW480, SW707, SW48, and HT29) with widely differing radiosensitivities. Cell survival curves were generated for a range of docetaxel concentrations (5-20 nM) alone and for X-rays (1-5 Gy) +/- 10 or 20 nM docetaxel (for 24 h before irradiation). Cell cycle distributions and apoptotic frequencies were measured during the treatments. Sensitivity to docetaxel alone was similar in all cell lines and could be attributed to massive induction of apoptosis (60-80% by 24 h). Radiosensitivity varied widely; the surviving fractions at 2 Gy in the most resistant (HT29) and most sensitive (SW28) lines were 0.81 and 0.13, respectively. Exposure to 10 nM docetaxel induced a progressive accumulation of SW480, SW707, and SW48 cells in G2/M. After 24 h, 55-70% of the cells were in G2/M. It is likely, therefore, that accumulation in this radiosensitive phase of the cell cycle contributes significantly to radiosensitization by the drug.


Determinants of 6-month mortality in survivors of myocardial infarction after thrombolysis. Results of the GISSI-2 data base. The Ad hoc Working Group of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardico (GISSI)-2 Data Base.
Current knowledge of risk assessment in survivors of myocardial infarction is largely based on data gathered before the advent of thrombolysis. It must be determined whether and to what extent available information and proposed criteria of prognostication are applicable in the thrombolytic era. We reassessed risk prediction in the 10,219 survivors of myocardial infarction with follow-up data available (ie, 98% of the total) who had been enrolled in the GISSI-2 trial, relying on a set of prespecified variables. The 3.5% 6-month all-cause mortality rate of these patients compared with the higher value of 4.6% found in the corresponding GISSI-1 cohort, originally allocated to streptokinase therapy, indicates a 24% reduction in postdischarge 6-month mortality. On multivariate analysis (Cox model), the following variables were predictors of 6-month all-cause mortality: ineligibility for exercise test for both cardiac (relative risk [RR], 3.30; 95% confidence interval [CI], 2.36-4.62) and noncardiac reasons (RR, 3.28; 95% CI, 2.23-4.72), early left ventricular failure (RR, 2.41; 95% CI, 1.87-3.09), echocardiographic evidence of recovery phase left ventricular dysfunction (RR, 2.30; 95% CI, 1.78-2.98), advanced (more than 70 years) age (RR, 1.81; 95% CI, 1.43-2.30), electrical instability (ie, frequent and/or complex ventricular arrhythmias) (RR, 1.70; 95% CI, 1.32-2.19), late left ventricular failure (RR, 1.54; 95% CI, 1.17-2.03), previous myocardial infarction (RR, 1.47; 95% CI, 1.14-1.89), and a history of treated hypertension (RR, 1.32; 95% CI, 1.05-1.65). Early post-myocardial infarction angina, a positive exercise test, female sex, history of angina, history of insulin-dependent diabetes, and anterior site of myocardial infarction were not risk predictors. On further multivariate analysis, performed on 8315 patients with the echocardiographic indicator of left ventricular dysfunction available, only previous myocardial infarction was not retained as an independent risk predictor. A decline in 6-month mortality of myocardial infarction survivors, seen within 6 hours of symptom onset, has been observed in recent years. Ineligibility for exercise test, early left ventricular failure, and recovery-phase left ventricular dysfunction are the most powerful (RR, > 2) predictors of 6-month mortality among patients recovering from myocardial infarction after thrombolysis. Qualitative variables reflecting residual myocardial ischemia do not appear to be risk predictors. The lack of an independent adverse influence of early post-myocardial infarction angina on 6-month survival represents a major difference between this study and those of the prethrombolytic era.


Clinical and safety outcomes associated with treatment of acute venous thromboembolism: a systematic review and meta-analysis.
Many anticoagulant strategies are available for the treatment of acute venous thromboembolism, yet little guidance exists regarding which drug is most effective and safe. To summarize and compare the efficacy and safety outcomes associated with 8 anticoagulation options (unfractionated heparin [UFH], low-molecular-weight heparin [LMWH], or fondaparinux in combination with vitamin K antagonists); LMWH with dabigatran or edoxaban; rivaroxaban; apixaban; and LMWH alone) for treatment of venous thromboembolism. A systematic literature search was conducted using MEDLINE, EMBASE, and the evidence-based medicine reviews from inception through February 28, 2014. Eligible studies were randomized trials reporting rates of recurrent venous thromboembolism and major bleeding in patients with acute venous thromboembolism. Of the 1197 studies identified, 45 trials including 44,989 patients were included in the analyses. Two reviewers independently extracted trial-level data including number of patients, duration of follow-up, and outcomes. The data were pooled using network meta-analysis. The primary clinical and safety outcomes were recurrent venous thromboembolism and major bleeding, respectively. Compared with the LMWH-vitamin K antagonist combination, a treatment strategy using the UFH-vitamin K antagonist combination was associated with an increased risk of recurrent venous thromboembolism (hazard ratio [HR], 1.42; 95% credible interval [CrI], 1.15-1.79). The proportion of patients experiencing recurrent venous thromboembolism during 3 months of treatment were 1.84% (95% CrI, 1.33%-2.51%) for the UFH-vitamin K antagonist combination and 1.30% (95% CrI, 1.02%-1.62%) for the LMWH-vitamin K antagonist combination. Rivaroxaban (HR, 0.55; 95% CrI, 0.35-0.89) and apixaban (HR, 0.31; 95% CrI, 0.15-0.62) were associated with a lower risk of bleeding than was the LMWH-vitamin K antagonist combination, with a lower proportion of patients experiencing a major bleeding event during 3 months of anticoagulation: 0.49% (95% CrI, 0.29%-0.85%) for rivaroxaban, 0.28% (95% CrI, 0.14%-0.50%) for apixaban, and 0.89% (95% CrI, 0.66%-1.16%) for the LMWH-vitamin K antagonist combination. Using meta-analytic pooling, there were no statistically significant differences for efficacy and safety associated with most treatment strategies used to treat acute venous thromboembolism compared with the LMWH-vitamin K antagonist combination. However, findings suggest that the UFH-vitamin K antagonist combination is associated with the least effective strategy and that rivaroxaban and apixaban may be associated with the lowest risk for bleeding.


Long term effects of temperature and substrate level on BNR with an external nitrification reactor.
This study was conducted with a BNR (biological nutrient removal) process with an external nitrification and switching arrangement of anoxic and oxic stages. It was observed that the SPRR (specific phosphorus release rate) and SDNR (specific denitrification rate) were greatly affected by the organic loads, and SDNR had a higher temperature effect than other kinetic rates including nitrification. It was further observed that the stoichiometric values like PHA (poly hydroxyalkanoate) stored for P release and PHA consumed for P uptake also varied. Variations of % Px (phosphorus content) and PHA as intracellular matter suggest the PAOs were more active at lower temperatures with this process configuration, where more than 70% of NH4-N and phosphorus were removed at temperatures below 10 degrees.


Do People Believe That They Are More Deontological Than Others?
The question of how we decide that someone else has done something wrong is at the heart of moral psychology. Little work has been done to investigate whether people believe that others' moral judgment differs from their own in moral dilemmas. We conducted four experiments using various measures and diverse samples to demonstrate the self-other discrepancy in moral judgment. We found that (a) people were more deontological when they made moral judgments themselves than when they judged a stranger (Studies 1-4) and (b) a protected values (PVs) account outperformed an emotion account and a construal-level theory account in explaining this self-other discrepancy (Studies 3 and 4). We argued that the self-other discrepancy in moral judgment may serve as a protective mechanism co-evolving alongside the social exchange mechanism and may contribute to better understanding the obstacles preventing people from cooperation.


Integrating Family Planning and HIV Services at the Community Level: Formative Assessment with Village Health Teams in Uganda.
Little is known on integrating HIV and family planning (FP) services in community settings. Using a cluster randomized controlled design, we conducted a formative assessment in two districts in Uganda where community health workers, called VHTs, already offered FP. Thirty-six trained VHTs also provided HIV testing and counseling (HTC) during the intervention. We surveyed all 36 VHTs and 256 FP clients, and reviewed service statistics. In the intervention group, VHTs tested 80% of surveyed clients for HIV, including 76% they already saw for FP and 22% who first came to them for HTC before receiving FP. Comparing clients' experiences in the intervention and control groups, adding HTC does not appear to have negatively affected FP service quality. VHTs reported more monthly clients, but rated their workload as easy to manage. This integrated model seems feasible and beneficial for both VHTs and clients, while not resulting in any negative effects. This study was registered with ClinicalTrials.gov, number [NCT02244398].


Chloroplast Differentiation in Tomato Root Galls Induced by the Root Knot Nematode Meloidogyne incognita.
Primary roots of tomato, Lycopersicon esculentum cv. Marglobe, were cultured aseptically on agar containing a standard nutrient formulation with or without kinetin. When secondary roots developed, cultures were inoculated with the root-knot nematode, Meloidogyne incognita. Following inoculation, the cultures were divided into two groups which were incubated either in total darkness or in 16-h light-8-h dark cycles. At 24 h, 1, 2, 3, and 4 wk after incubation, roots from all cultures were processed for transmission electron microscopy. Fine structural observation of the parenchyma tissue in galls from the inoculated cultures indicated that starch containing plastids or amyloplasts, which are usually present and remain undifferentiated in these root cells, developed into chloroplasts. These chloroplasts contained a membrane system indistinguishable from those found in leaves of intact plants. Although plastid development was not affected when uninoculated cultures were incubated in the light, differentiation of the amyloplast was induced when roots were cultured on the medium containing kinetin. These results suggest that plastid differentiation in the inoculated tissue may be influenced by an accumulation of kinetin in the gall, which is induced by the nematode and serves as the nutrient sink for its feeding.


[Otalgia and impairment of hearing in the diagnosis of inflammatory and tumorous diseases of the pharynx and larynx].
Otalgia and/or impairment of hearing is one of the earliest, if not the first, symptom of certain pharyngeal and laryngeal disorders. Underestimation of these conditions or incorrect interpretation of their origin frequently lead to erroneous diagnosis and the wrong choice of treatment strategy. A total of 1074 patients with pharyngeal and laryngeal problems were available for examination that revealed pathologies in which otalgia and/or impairment of hearing is the first or an earlier symptom of the disease. Otalgia proved to be an early manifestation of serious inflammatory diseases affecting the middle part of the pharynx or of metastasis of malignant pharyngeal and laryngeal tumours into deep cervical lymph nodes. Juvenile angiofibroma, hemangiouma, malignant nasopharyngeal tumour are known to disturb functional automatism of pharyngeal openings of Eustachian tubes and cause impairment of hearing long before clinical manifestations of neoplastic growth. Results of diagnostic studies are usually interpreted as middle ear pathology (tubootitis, exudative otitis, middle ear inflammation). Correct diagnosis is possible within 1-12 months (or more) after the appearance of the first symptoms. An error is possible to avoid by thorough examination of the pharynx using endoscopic optics. Target biopsy is indicated as soon as neoplastic microlesions are identified excepting cases of branched arterial-type hemangiomas.


Temporal overlap and repeatability of feather corticosterone levels: practical considerations for use as a biomarker.
The measurement of corticosterone (CORT) levels in feathers has recently become an appealing tool for the conservation toolbox, potentially providing a non-invasive, integrated measure of stress activity throughout the time of feather growth. However, because the mechanism of CORT deposition, storage and stability in feathers is not fully understood, it is unclear how reliable this measure may be, especially when there is an extended interval between growth and feather collection. We compared CORT levels of naturally grown feathers from tree swallows (Tachycineta bicolor) that were moulted and regrown concurrently and therefore expected to have similar CORT levels. Specifically, we compared the same feather from the left and right wing (moulted symmetrically) and different types of feathers (wing, back and tail) expected to have been moulted within the same time period. We found that larger, heavier feathers held more CORT per unit length. In addition, we found a lack of concordance in CORT levels both within the same feather type and between different feather types, even after taking into account differences in feather density. Our results indicate that naturally grown feathers may not consistently provide an indication of stress status. Additionally, conflict in results may arise depending on the feather assayed, and total feather volume may be an important consideration when interpreting feather CORT levels. Future work is necessary to determine explicitly the mechanisms of CORT deposition, the effects of environmental exposure and feather wear on the permanence of the feather CORT signal, and the influence of responses to wild stressors on feather CORT levels, before feather CORT can be implemented effectively as a tool for ecological and conservation applications.


Loss of lymphocyte cyclic AMP dependent protein kinase activity in malignant melanoma.
Cyclic AMP dependent protein kinase activity was depressed in whole thymus and spleen as well as isolated splenic lymphocytes from B16 melanoma bearing C57B1/6J mice as compared to control animals. A similar loss of enzyme activity was observed in human peripheral blood lymphocytes from melanoma bearing patients as compared to normal subjects. An unaltered level of activity in the heart of tumor bearing mice suggested some specificity for the lymphoid system. This depressed enzyme activity was the result of a diminished Vmax for cAMP stimulated calf histone phosphorylation. The tumor bearing state in the mouse was also accompanied by a depletion of small lymphocytes from both thymus and spleen and it is hypothesized that the losses of lymphocytes and cAMP dependent protein kinase activity are related.


Direct role of furin in mammalian prosomatostatin processing.
We have previously reported that rat prosomatostatin (rPSS) undergoes conversion at Arg decreases and Lys decreases monobasic sites to SS-28 and PSS-(1-10) respectively in COS-7 cells, and have proposed furin or a related enzyme of the constitutive secretory pathway as the endoproteinase responsible. Here we have tested directly the ability of furin to cleave rPSS at the two monobasic sites as well as at the RXRK dibasic site of SS-14 conversion (a furin motif, except for Lys substituting for Arg at P1). Recombinant vaccinia virus (VV) vectors were used to co-express rPSS with graded doses of furin in COS-7 cells and LoVo colon carcinoma cells deficient in furin. PSS and cleavage products in cell extracts and media were characterized by HPLC analysis and C-terminal [SS-14-like immunoreactivity (SS-14 LI)] and N-terminal [PSS-(1-10) LI] directed radioimmunoassays. There was a dose-dependent increase in SS-28 production from rPSS by furin in COS-7 cells from 29% (control) to 58% (high-dose furin) associated with a progressive decrease in unprocessed PSS from > 60% to approximately 20% of total SS-14 LI. Significant SS-14 production occurred only at high levels of furin infection. Control LoVo cells infected with VV:rPSS exhibited production of approximately 21% SS-28, approximately 15% PSS-(1-10) and 3.5% SS-14. Infection of LoVo cells with VV:hfurin (hfurin = human furin) enhanced SS-28 production to 30-34%. SS-14 synthesis also increased to 25-40%, probably by conversion from SS-28. Overexpression of furin in COS-7 or LoVo cells failed to increase PSS-(1-10) production. These results show that furin is a candidate SS-28 convertase. Arginine is the preferred residue at the P1 site of furin cleavage. Furin does not process rPSS to PSS-(1-10), suggesting the existence of another monobasic convertase with a preference for Lys rather than Arg at P1. Such an enzyme could also explain the presence of endogenous SS-28-, PSS-(1-10)- and SS-14-producing activities in LoVo cells.


Clinico-pathological profile of Hairy cell leukemia: critical insights gained at a tertiary care cancer hospital.
Hairy cell leukemia (HCL) is a rare, low grade, B-cell neoplasm with a characteristic morphologic and immunophenotypic profile. It has to be distinguished from chronic lymphoproliferative disorders because of different treatment protocol and clinical course. To evaluate clinicopathological features including immunophenotypic analysis of cases diagnosed as HCL. The present study included 28 cases diagnosed over a period of nine years (2002-2010). Clinical presentation, complete blood count, bone marrow aspirate, and flow cytometric analysis of cases were reviewed. Treatment and follow-up details (ranging from 3-90 months) were noted. This study revealed 28 cases (referrals-7, indoor-21), aged 26-69 years with a median age of 47 years, with a male predominance (M:F=6:1). The presenting complaints were weakness (80%) followed by fever (56%) and abdominal pain. Physical examination revealed splenomegaly in most patients (92%) and hepatomegaly in a minority (28%). The common laboratory features were anemia in 23 cases, pancytopenia in 14 cases, while two patients had leukocytosis and three patients had normal WBC count. Dry tap was observed in 84% of the cases where hairy cells constituted 16-97% of non-erythroid nucleated cells. Tartarte resistant acid phosphate staining was positive in all the eight cases where it was done. CD5 was negative in all the cases, while CD10 was expressed in three cases (13%) and CD23 in five cases (19%). Though pancytopenia is common, occasional patient can present with normal blood counts or leukocytosis. Few unusual findings include presence of lymphadenopathy, absence of palpable splenomegaly, and expression of CD23 and CD10 by the leukemic cells.


Plant foods, fiber, and rectal cancer.
Associations between colon and rectal cancer and intakes of vegetables, other plant foods, and fiber have stimulated much debate. We examined the association between rectal cancer and plant food and fiber intakes. Data from 952 incident cases of rectal cancer were compared with data from 1205 population-based controls living in Utah or enrolled in the Kaiser Permanente Medical Care Program in northern California Rectal cancer was inversely associated with intakes of vegetables (odds ratio: 0.72; 95% CI: 0.54, 0.98), fruit (0.73; 0.53, 0.99), and whole-grain products (0.69; 0.51, 0.94), whereas a high intake of refined-grain products was directly associated with an increased risk of rectal cancer (1.42; 1.04, 1.92). Similarly, relative to low fiber intakes, high intakes of dietary fiber reduced the risk of rectal cancer (0.54; 0.37, 0.78). The reduced risk of rectal cancer associated with vegetable (0.48; 0.29, 0.80), fruit (0.63; 0.38, 1.06), and fiber (0.40; 0.22, 0.71) intakes was strongest for persons who received the diagnosis after age 65 y. A threshold effect at approximately 5 servings of vegetables/d was needed to see a reduced risk of rectal cancer. The results suggest that plant foods may be important in the etiology of rectal cancer in both men and women. Age at diagnosis appears to play an important role in the association.


Arabidopsis GAAP1 to GAAP3 Play Redundant Role in Cell Death Inhibition by Suppressing the Upregulation of Salicylic Acid Pathway Under Endoplasmic Reticulum Stress.
The unfolded protein response (UPR) is activated to sustain cell survival by reducing misfolded protein accumulation in the endoplasmic reticulum (ER). The UPR also promotes cell death when the ER stress is severe. However, the underlying molecular mechanisms of UPR activity regulation and cell death transition are less understood in plants. Arabidopsis GAAP1 and GAAP3 are involved in the regulation of UPR and cell death. Five GAAP gene members are found in Arabidopsis. Here, we analyzed the function of GAAP2 in addition to GAAP1 and GAAP3 in ER stress response using single, double, and triple mutants. Results showed that single or double or triple mutants reduced plant survival and enhanced cell death under ER stress. And the sensitivity increased with the number of mutation genes increase. Quantitative real-time polymerase chain reaction analysis showed that mutation in triple genes promoted UPR signaling when confronted with mild ER stress, advanced SA target genes upregulation when confronted with severe stress. Moreover, Quantitative detection by UPLC-ESI-MS/MS showed that ER stress upregulated salicylic acid (SA) content in plants. These data suggest that GAAP1 to GAAP3 played redundant roles in cell death resistance and fine tuning UPR activation. And the anti-cell death function of GAAPs might be achieved by impairing the up-regulation of the SA pathway under ER stress.


Effect of topography-dependent light coupling through a near-field aperture on the local photocurrent of a solar cell.
An aperture-type scanning near-field optical microscope (a-SNOM) is readily used for the optical and optoelectronic characterizations of a wide variety of chemical, biological and optoelectronic samples with sub-wavelength optical resolution. These samples mostly exhibit nanoscale topographic variations, which are related to local material inhomogeneity probed either by an optical contrast or by secondary effects such as photoconductivity or photoluminescence. To date, in the interpretation and evaluation of the measurement results from a-SNOM or derived methods, often only the local material inhomogeneity is taken into account. A possible influence of the optical interaction between the scanning probe and the surface topography is rarely discussed. In this paper, we present experimental and theoretical investigation of the effects of nanoscale topographic features on a-SNOM measurement results. We conduct local photocurrent measurements on a thin-film solar cell with an a-SNOM as the illumination source. A clear correlation between the photocurrent response and local topography is observed in all measurements with a signal contrast of up to ∼30%, although the sample features homogeneous permittivity and electrical properties. With the help of finite-difference time-domain (FDTD) simulations, this correlation is reproduced and local light coupling is identified as the mechanism which determines the local photocurrent response. Our results suggest that a-SNOM-based measurements of any sample with material inhomogeneity will be superimposed by the local light-coupling effect if surface topography variation exists. This effect should always be taken into consideration for an accurate interpretation of the measurement results.


Resistance to differentiation affects ribo- and deoxyribonucleotide pools and sensitivity to pyrimidine metabolism antagonists in HL60 cells.
HL60 myeloid leukemia cells are extensively used as a differentiation model. We investigated a variant of HL60 which is resistant to differentiation induction (HL60-R) by standard differentiation inducers such as retinoic acid and dimethylsulfoxide (DMSO). To find an explanation for this resistance, we examined nucleotide (NTP) and deoxynucleotide (dNTP) pools in HL60-R and its parent cell line, sensitive to differentiation, HL60-S. We also explored whether these differences led to a difference in sensitivity to various antimetabolites. Drug sensitivity was measured with the tetrazolium (MTT) assay, while nucleotides were measured with anion-exchange HPLC. HL60-R cells were between 2- and 5-fold resistant to the antimetabolites 5-fluorouracil, Brequinar, hydroxyurea and N-(phosphonacetyl)-L-aspartate (PALA), but more sensitive to aza-2'-deoxycytidine (DAC), cytarabine and thymidine (5- to 10-fold). The NTP pools in both HL60 variants showed a normal pattern with ATP being the highest (2530-2876 pmol/106 cells) and CTP being lowest. However, UTP pools were 2-fold higher in the HL60-S cells (p < .01), while CTP and GTP pools were 30% higher (p < .01) compared to HL60-R cells. For the dNTP pools, larger differences were observed, with dATP (50 pmol/106 cells) being highest in HL60-R cells, but dATP was 4-fold lower in HL60-S cells. In HL-60-R, the triple combination retinoic acid, DMSO and DAC increased all NTPs almost 2-fold in contrast to HL60-S. Uridine increased UTP (1.4-fold), CTP (2-fold) and dCTP (1.4.-fold) pools in both cell lines, but thymidine increased only dTTP pools (4- to 7-fold), with a depletion of dCTP. PALA decreased UTP and CTP in both cell lines, but increased ATP (only in HL60-R). Hydroxyurea decreased dNTP especially in HL60-S cells. In conclusion, the pronounced differences in NTP and dNTP pools between HL60-S and HL60-R possibly play a role in the induction of differentiation and drug sensitivity.


Calcium regulates the production of nodulation outer proteins (Nops) and precludes pili formation by Sinorhizobium fredii USDA257, a soybean symbiont.
Sinorhizobium fredii USDA257 forms nitrogen-fixing nodules on primitive soybean cultivars such as 'Peking' but is unable to establish efficient symbiosis with North American cultivars. USDA257 when grown in presence of genistein, a potent nodD-inducing isoflavonoid, secretes at least six nodulation outer proteins (NopX, NopB, NopL, NopP, NopA and NopC) to the extracellular milieu through a type III secretion system. These proteins regulate legume nodulation in a host-specific manner. Here, it is demonstrated that calcium prevents the accumulation of NopB and NopA, and drastically reduces that of NopX and NopL. The inhibitory effect on Nops accumulation appears to be mediated specifically by calcium since other divalent cations such as Mg(2+) and Mn(2+) had no detectable effect. Calcium does not appear to interfere with the secretion of these proteins since Western blot analysis revealed that these Nops do not accumulate inside the cell. The inhibitory effect of calcium on Nops production is mediated at the posttranscriptional level. Studies by the authors indicate that the production of Nops, which function as determinants of host-range, is regulated by calcium.


Racial and ethnic differences in diabetes care and health care use and costs.
Previous studies have shown racial and ethnic differences in diabetes complication rates and diabetes control. The objective of this study was to examine racial and ethnic differences in diabetes care and health care use and costs for adults with diabetes using a nationally representative sample of the U.S. noninstitutionalized civilian population. We performed a cross-sectional analysis of the 2000 Medical Expenditure Panel Survey (MEPS) and its related Diabetes Care Survey. The respondents were adults (aged 18 years and older) with diabetes, including non-Hispanic whites, non-Hispanic African Americans, and Hispanics. Racial and ethnic differences were examined in diabetes process of care and health care use and costs using logistic regression, negative binomial regression, and ordinary least squares regression with log cost. Most of the outcomes in diabetes care management, treatment, and complications were not significantly different among race groups. After adjusting for socioeconomic and demographic characteristics, Hispanics were more likely to have eye problems than whites (odds ratio, 1.56; 95% confidence interval, 1.03-2.56). African Americans and Hispanics had lower total health care costs, lower ambulatory care costs, and lower prescription drug costs than whites (P < .01 for all). We found differences in ambulatory care and prescription drug fills among white, African American, and Hispanic adults with diabetes. However, most of the diabetes care measures were not significantly different among the three racial and ethnic groups. Understanding the reason outcomes do not differ when health care use and costs differ significantly should be a focus of future studies.


Independent control processes? Evidence for concurrent distractor inhibition and attentional usage of distractor information.
Interference evoked by a distractor presented prior to a target stimulus is reduced when the distractor-target SOA is increased, suggesting inhibition of distractor-related activation. Distractor processing is also assumed to be (strategically) adjusted to the proportions of congruent and incongruent target-distractor combinations, yielding a larger distractor interference effect when the proportion of congruent trials is higher (i.e., Proportion Congruent Effect, PCE). To explore the interplay of proportion congruent-based processing adjustment and the time course of distractor-related activation we varied the proportions of congruent and incongruent trials as well as the distractor-target SOA. To control for item-specific priming we kept distractor-related contingencies (i.e., frequency of individual distractor-target conjunctions) constant for a subset of the stimuli (and used a different subset to manipulate the proportions of congruent and incongruent trials). A PCE occurred, even for the subset of stimuli associated with constant distractor-related contingencies, thus ruling out item-specific contingency learning. Distractor interference was reduced when the SOA was increased, but this reduction did not differ between the proportion congruent conditions, as confirmed by a Bayesian analysis. Our results are consistent with independent processes pertaining to usage of distractor information for biasing response selection and distractor inhibition during the SOA. Alternative interpretations of the independent effects of the PC manipulation and the distractor-target SOA are discussed.


Expression of the murine CD27 ligand CD70 in vitro and in vivo.
The interaction between TNFR family member CD27 and its ligand CD70 promotes lymphocyte expansion and effector cell formation. In humans, control of CD27 function is partly regulated by the restricted expression of CD70. We used newly developed mAbs to characterize murine (m) CD70 expression in vitro and in vivo. On resting lymphocytes and immature dendritic cells (DC), mCD70 is absent. In vitro, Ag receptor triggering induced mCD70 mRNA in T cells, but cell surface protein expression was very low. Activated B cells synthesized much higher levels of mCD70 mRNA than activated T cells and clearly expressed mCD70 at the cell surface. mCD70 cell surface expression could also be induced on the DC line D1 and on in vitro-generated murine DC upon maturation. In lymphoid organs of naive mice, virtually no mCD70-expressing cells were found, with exception of cells in the thymic medulla, which may be epithelial in origin. However, after intranasal infection with influenza virus, lung-infiltrating T cells and T and B cells in draining lymph nodes expressed mCD70 according to immunohistology. In such activated lymphocytes, mCD70 protein is largely retained intracellularly. Plasma membrane expression of mCD70 was only detectable by flow cytometry on a small proportion of lung-infiltrating T cells and peaked at the height of the primary response. Thus, expression of CD70 in the mouse is highly regulated at the transcriptional and posttranslational level. This most likely serves to limit excessive effector cell formation after antigenic stimulation.


HOXA7 stimulates human hepatocellular carcinoma proliferation through cyclin E1/CDK2.
HOX genes are transcription factors that control morphogenesis, organogenesis and differentiation. Increasing evidence suggests that HOX genes play a role in hepatocellular carcinoma (HCC) progression; however few studies have defined the functional roles and mechanisms of action. In the present study, we used siRNA and forced-expression in multiple cell lines to define the role of HOXA7 in the regulation of proliferation of HCC in vitro and in vivo. Knockdown of endogenous HOXA7 decreased the proliferation of HepG2 and QGY-7703 cells. These changes were not associated with significant changes in cyclin D1 and CDK4. However, downregulation of HOXA7 significantly reduced cyclin E1 and CDK2 protein levels. Conversely, overexpression of HOXA7 in QSG-7701 cells stimulated proliferation and increased cyclin E1 and CDK2 protein levels. Our results confirmed that HOXA7 promoted cell proliferation, and these changes were mediated by cyclin E1/CDK2. These observations contribute to our understanding of the important roles of HOXA7 in HCC development and progression and HOXA7 could be a promising molecular target for the development of new diagnostic and therapeutic strategies for HCC.


[Liver injury due to COVID-19 in critically ill adult patients. A retrospective study].
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged, causing the current pandemic of acute respiratory disease known as COVID-19. Liver injury due to COVID-19 is defined as any liver injury occurring during the course of the disease and treatment of patients with COVID-19, with or without liver disease. The incidence of elevated liver transaminases, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), ranges from 2.5 to 76.3%. The aim of the present study was to describe the hepatic biochemical abnormalities, after a SARS-CoV-2-positive polymerase chain reaction (PCR) test, and the mortality rate in critically ill patients. A retrospective study was conducted that included 70 patients seen at a private hospital in Mexico City, within the time frame of February-December 2021. Median patient age was 44.5 years (range: 37-57.2) and 43 (61.4%) of the patients were men. Liver function tests were performed on the patients at hospital admission. Gamma glutamyl transferase (GGT) levels were elevated (p = 0.032), as were those of AST (p = 0.011) and ALT (p = 0.021). The patients were stratified into age groups: 18-35, 36-50, and > 50 years of age. The 18 to 35-year-olds had the highest liver enzyme levels and transaminase levels were higher, the younger the patient. Due to the low mortality rate (one patient whose death did not coincide with a hepatic cause), the multivariate analysis showed an R2 association of 0.689, explained by AST, GGT, and C-reactive protein levels. Despite the increase in transaminases in our study population during the course of COVID-19, there was no increase in mortality. Nevertheless, hospitalized patient progression should be continuously followed.


Atypical Sensory behaviours in children with Tourette's Syndrome and in children with Autism Spectrum Disorders.
Certain visual disturbances make it difficult to read text and have been attributed to visual stress, also called "pattern-related visual stress". 12 Children with ASD, 12 children with Tourette's syndrome and without ASD and 12 controls, all matched on age and non verbal ability, participated in an experiment exploring sensory behaviours and visual stress. Reading rate and accuracy were assessed with the Wilkins Rate of Reading test with and without the Intuitive Overlays. Both the children with Tourette's and the children with ASD showed a higher prevalence of atypical sensory behaviours and symptoms of visual stress than the typically developing control children. Six out of twelve children with Tourette's syndrome (50%) read more accurately and over 15% more quickly with a coloured overlay. Four of the 12 children with ASD and none of the control children read over 15% more quickly with an overlay. The findings are discussed in relation to problems in sensory modulation.


Effect of once-weekly dulaglutide versus insulin glargine in people with type 2 diabetes and different baseline glycaemic patterns: A post hoc analysis of the AWARD-2 clinical trial.
The long-acting glucagon-like peptide-1 receptor agonist dulaglutide acts by stimulating insulin secretion and reducing glucagon levels in a glucose-dependent manner both in the fasting and postprandial states, resulting in reductions of both fasting glucose (FG) and postprandial glucose (PPG). In contrast, the main mechanism of action of basal insulin is to reduce elevated FG by inhibiting hepatic glucose production. The aim of the present post hoc analysis of the phase 3 AWARD-2 trial was to investigate whether specific baseline glycaemic patterns respond differentially to dulaglutide compared to insulin glargine (glargine). We categorized participants into four subgroups based on prespecified glucose thresholds and their baseline FG and daily 2-hour mean PPG: low FG/low PPG; low FG/high PPG; high FG/low PPG; and high FG/high PPG. Changes in glycaemic measures in response to treatment with dulaglutide or glargine were evaluated in each subgroup. At 52 weeks, significant reductions from baseline in glycated haemoglobin (HbA1c) were observed in all subgroups with dulaglutide 1.5 mg and with glargine (all P < .05), except in patients with low FG/low PPG who received glargine. Greater HbA1c reductions were observed with dulaglutide 1.5 mg compared to glargine in all subgroups (all P ≤ .05), except in the low FG/high PPG subgroup.


Incidence of DWI-positive stroke in patients with vertigo of unclear etiology, preliminary results.
Acute vertigo may be secondary to stroke or to non-ischemic causes. Accurate identification of vertigo secondary to ischemia may lead to appropriate timely intervention that can minimize stroke-related damage and can help in tailoring the most appropriate individual therapy for affected patients. Diffusion weighted MRI (DWI) is very accurate for diagnosing stroke and we therefore aimed to test whether it can aid in making a correct diagnosis of vertigo secondary to stroke. All patients presenting with vertigo in which the diagnosis of stroke was considered underwent DWI only MRI. Data regarding the symptoms and neurological deficits, vascular risk factors, imaging findings, and outcomes was accrued. Patients with stroke on DWI were compared with those without ischemia. Between June 2010 and August 2011, 28 patients fulfilling the entry criteria were identified with a mean age of 62·2±12·8 (60% male). The final diagnosis was stroke in 11 patients (39%). Patients with stroke did not differ from those without stroke in their risk factor profile. However, patients with stroke more often tended to present with vertigo accompanied by other neurological symptoms (73% versus 12% respectively, P = 0·001). After adjusting for age and the presence of diabetes, the presence of multiple symptoms remained the only variable that was associated with a positive DWI scan (odds ratio: 30: 95% confidence interval: 2·6-349). Most patients with stroke had very mild strokes with a median admission NIHSS score of 3 and DWI lesion volumes >2 cm were found in only three patients. Most stroke patients made a good recovery (modified Rankin score ≤2 in seven of nine patients with 90 day data). The most common diagnosis in patients without stroke was of vertigo of peripheral origin (14/17). DWI only MRI can be used to rapidly screen patients presenting with vertigo and suspected vertebrobasilar stroke. The occurrence of vertigo in combination with other focal neurological symptoms may increase the accuracy of stroke diagnosis but is not sensitive or specific enough. Combining clinical presentation patterns with DWI data may enable rapid decision making as to treatment options.


PRSice-2: Polygenic Risk Score software for biobank-scale data.
Polygenic risk score (PRS) analyses have become an integral part of biomedical research, exploited to gain insights into shared aetiology among traits, to control for genomic profile in experimental studies, and to strengthen causal inference, among a range of applications. Substantial efforts are now devoted to biobank projects to collect large genetic and phenotypic data, providing unprecedented opportunity for genetic discovery and applications. To process the large-scale data provided by such biobank resources, highly efficient and scalable methods and software are required. Here we introduce PRSice-2, an efficient and scalable software program for automating and simplifying PRS analyses on large-scale data. PRSice-2 handles both genotyped and imputed data, provides empirical association P-values free from inflation due to overfitting, supports different inheritance models, and can evaluate multiple continuous and binary target traits simultaneously. We demonstrate that PRSice-2 is dramatically faster and more memory-efficient than PRSice-1 and alternative PRS software, LDpred and lassosum, while having comparable predictive power. PRSice-2's combination of efficiency and power will be increasingly important as data sizes grow and as the applications of PRS become more sophisticated, e.g., when incorporated into high-dimensional or gene set-based analyses. PRSice-2 is written in C++, with an R script for plotting, and is freely available for download from http://PRSice.info.


What makes rhythms hard to perform? An investigation using Steve Reich's Clapping Music.
Clapping Music is a minimalist work by Steve Reich based on twelve phased variations of a rhythmic pattern. It has been reimagined as a game-based mobile application, designed with a dual purpose. First, to introduce new audiences to the Minimalist genre through interaction with the piece presented as an engaging game. Second, to use large-scale data collection within the app to address research questions about the factors determining rhythm production performance. The twelve patterns can be differentiated using existing theories of rhythmic complexity. Using performance indicators from the game such as tap accuracy we can determine which patterns players found most challenging and so assess hypotheses from theoretical models with empirical evidence. The app has been downloaded over 140,000 times since the launch in July 2015, and over 46 million rows of gameplay data have been collected, requiring a big data approach to analysis. The results shed light on the rhythmic factors contributing to performance difficulty and show that the effect of making a transition from one pattern to the next is as significant, in terms of pattern difficulty, as the inherent complexity of the pattern itself. Challenges that arose in applying this novel approach are discussed.


Swedish version of the multi dimensional health assessment questionnaire -- translation and psychometric evaluation.
Health assessment measurements for patients with Rheumatoid arthritis (RA) have to be meaningful, valid and relevant. A commonly used questionnaire for patients with RA is the Stanford Health Assessment Questionnaire Disability Index (HAQ), which has been available in Swedish since 1988. The HAQ has been revised and improved several times and the latest version is the Multi Dimensional Health Assessment Questionnaire (MDHAQ). The aim of this study was to translate the MDHAQ to Swedish conditions and to test the validity and reliability of this version for persons with RA. Translation and adaption of the MDHAQ were performed according to guidelines by Guillemin et al. The translated version was tested for face validity and test-retest in a group of 30 patients with RA. Content validity, criterion validity and internal consistency were tested in a larger study group of 83 patients with RA. Reliability was tested with test-retest and Cronbach´s alpha for internal consistency. Two aspects of validity were explored: content and criterion validity. Content validity was tested with a content validity index.Criterion validity was tested with concurrent validity by exploring the correlation between the MDHAQ-S and the AIMS2-SF. Floor and ceiling effects were explored. Test-retest with intra-class correlation coefficient (ICC) gave a coefficient of 0.85 for physical function and 0.79 for psychological properties. Reliability test with Cronbach´s alpha gave an alpha of 0.65 for the psychological dimension and an alpha of 0.88 for the physical dimension of the MDHAQ-S.The average sum of the content validity index for each item was of the MDHAQ-S was 0.94. The MDHAQ-S had mainly a moderate correlation with the AIMS2-SF, except for the social dimension of the AIMS2-SF, which had a very low correlation with the MDHAQ-S. The MDHAQ-S was considered to be reliable and valid, but further research is needed concerning sensitivity to change.


Rapid resolution of traumatic acute subdural haematoma in the elderly.
Rapid resolution of acute subdural haematoma (ASDH) has rarely been observed. Two elderly patients with rapid resolution of ASDH are described. In the present cases, it was indicated that a mechanism of rapid resolution of ASDH was attributable to cerebrospinal fluid dilution and cerebral atrophy resulting from aging.


Using 3-6 differences in essential fatty acids rather than 3/6 ratios gives useful food balance scores.
The vitamin-like omega-3 and omega-6 essential fatty acids are converted in the body to a large family of hormones which act at selective receptors that occur on nearly every cell and tissue. A relative omega-3 deficit allows overabundant actions of omega-6 hormones to develop into health disorders. People need simple, explicit information on the balance of essential fatty acids in their foods to avoid accumulating unintended imbalances in their tissue omega-3 and omega-6 fatty acids. We developed an Omega 3-6 Balance Food Score that summarizes in a single value the balance among eleven omega-3 and omega-6 essential fatty acids in a food. The value allows a quantitative estimate of the impact of each food item on the proportions of omega-3 and omega-6 that will accumulate in the 20- and 22-carbon highly unsaturated fatty acids of blood, which is an important health risk assessment biomarker. The impact of an individual food item upon a useful health risk assessment biomarker is easily evident in a simple, explicit value for the balance among eleven essential fatty acids nutrients. Foods with more positive Omega 3-6 Balance Food Scores will increase the percent of omega-3 in the biomarker, whereas those with more negative Scores will increase the percent of omega-6 in the biomarker.


[Approach to population-based screening in the Czech Republic, methodology and first results of the personalised invitation of citizens to cancer screening programmes].
In January 2014, a programme of personalised invitations was launched in the Czech Republic, with the objective of inviting insured persons to cancer screening programmes; namely breast cancer screening and cervical cancer screening in women, and colorectal cancer screening both in women and men. This programme aims at strengthening the current cancer prevention programmes, and to increase the currently inadequate participation of the target population in these programmes; therefore, personalised invitations are sent to citizens who have not participated in these programmes for several years and therefore at risk of developing a serious disease. The project is coordinated by the Czech Ministry of Health in cooperation with the expert medical societies involved (gynaecology, gastroenterology, gastrointestinal oncology, diagnostic radiology, general practice), representatives of health care payers, and other experts nominated by the Minister of Health. All health care payers invite their clients (insured persons) to preventive check-ups, covering all examinations needed. The project has been realised with the assistance of financial resources from EU funds. This article describes the methodology of personalised invitations which has been implemented nationwide, its data background, and the first results of the project in the first half of 2014, when almost 1.3 million Czech citizens were invited.


How Serious Are Health-Related Welfare Problems in Unowned Unsocialised Domestic Cats? A Study from Denmark Based on 598 Necropsies.
Free ranging unsocialised domestic cats are widely believed to suffer from a high load of welfare problems. We assessed the validity of this belief by performing necropsies on the corpses of 598 unsocialised cats, originating from all parts of Denmark, that had been euthanised by two Danish cat welfare organisations. We selected a number of variables for health-related cat welfare that could be assessed through necropsy (e.g., gross lesions, ectoparasites and body condition) or by laboratory analysis (e.g., infection with feline immunodeficiency virus (FIV), and by feline leukaemia virus (FeLV)). Each finding was classified as having either a major or minor welfare impact on the cat. More than 83% of the cats had no major finding, and 54% had no finding indicating a welfare issue at all. More than 83% of the cats had a body condition within normal range. Only 0.3% were emaciated. The most common finding was infestation with ectoparasites, with 15.9% infected with lice, 12.3% with fleas, 4.7% with ticks, and 6.7% with ear mites. FIV and FeLV were detected in 9.2% and 1.2% of the cases, respectively. The most common lesion related to the cats' teeth. Overall, unsocialised cats in Denmark have a moderate level of health-related welfare problems.


Recovery in community: using participatory action research to explore recovery with alternatives.
In Ontario, many people are calling for recovery to become the overarching philosophy and goal of the mental health system. Mental health service organizations are key to promoting recovery, but relatively little has been done to understand what recovery means at the organizational level. This paper describes the context, conduct, and findings of a participatory action research project designed to explore recovery in Alternatives, an organization in Toronto providing mental health services.


Regulation of Smoothened Trafficking and Abundance in Hedgehog Signaling.
The GPCR-family protein Smoothened (Smo) is essential for Hedgehog (Hh) signal transduction in both insects and vertebrates. The regulation of subcellular localization and abundance of Smo is a critical step in Hh signaling. Recent studies have demonstrated that Smo is subjected to ubiquitination mediated by multiple E3 ubiquitin ligases, leading to Smo endocytosis and subsequent degradation through the proteasome- and lysosome-mediated pathways in Drosophila. Ubiquitination of Smo also promotes its ciliary exit in mammalian cells. Hh inhibits Smo ubiquitination by blocking E3 ligase recruitment and promoting Smo deubiquitination through the ubiquitin-specific protease 8 (USP8) in Drosophila. Inhibition of Smo ubiquitination by Hh promotes Smo cell surface accumulation in Drosophila and ciliary accumulation in mammalian cells. Interestingly, Hh also induces sumoylation of Smo in both Drosophila and mammalian cells, which promotes Smo cell surface/ciliary accumulation. This review focuses on how ubiquitination and sumoylation regulate Smo intracellular trafficking and abundance and how these processes are regulated by Hh.


Interaction of a 60-kilodalton D-mannose-containing salivary glycoprotein with type 1 fimbriae of Escherichia coli.
A 60-kilodalton glycoprotein previously isolated and purified from human saliva (J. B. Babu, E. H. Beachey, D. L. Hasty, and W. A. Simpson, Infect. Immun. 51: 405-413, 1986) was found to interact with type 1 fimbriae and prevent adhesion of type 1 fimbriated Escherichia coli to animal cells in a D-mannose-sensitive manner. Purified salivary glycoprotein agglutinated type 1 fimbriated E. coli and, at subagglutinating concentrations, blocked the ability of type 1 fimbriated E. coli to attach to human buccal epithelial cells or agglutinate guinea pig erythrocytes. Both interactions were inhibited by alpha-methyl-D-mannoside but not by alpha-methyl-D-glucoside. Complexing of the glycoprotein to type 1 fimbriae was demonstrated by molecular sieve chromatography and modified Western blots. When mixed with type 1 fimbriae, the radiolabeled salivary glycoprotein coeluted with type 1 fimbriae from a column of Sepharose 4B. When blotted from a sodium dodecyl sulfate gel to nitrocellulose sheets, the glycoprotein interacted directly with type 1 fimbriae applied to the blots. Both of the latter interactions also were blocked by alpha-methyl-D-mannoside but not by alpha-methyl-D-glucoside. Chemical modification of the glycoprotein with sodium metaperiodate abolished its ability to interact with isolated type 1 fimbriae or type 1 fimbriated E. coli. These results suggest that the carbohydrate moiety of the 60-kilodalton glycoprotein serves as a receptor for type 1 fimbriae in the oral cavity, and we postulate that the interaction may cause agglutination and early removal of E. coli, thereby preventing colonization by these organisms of oropharyngeal mucosae and dental tissues.


Gemcitabine potentiates cisplatin cytotoxicity and inhibits repair of cisplatin-DNA damage in ovarian cancer cell lines.
Synergistic cytotoxicity between cisplatin and the nucleoside analog gemcitabine was observed in a panel of cisplatin-sensitive (2008, A2780) and -resistant (2008/C13*5.25, A2780/CP70) human ovarian cell lines. Previous studies have suggested a role for DNA repair in the mechanism of synergy between the two drugs. We therefore further investigated the hypothesis that the synergistic cytotoxicity between gemcitabine and cisplatin in these cell lines may be caused by gemcitabine-mediated inhibition of cisplatin intrastrand adduct (IA) and interstand cross-link (ICL) repair. The effect of gemcitabine on the accumulation and repair of cisplatin IA and ICL in each cell line was then measured directly using gene-specific quantitative polymerase chain reaction and denaturation/renaturation techniques, respectively. Pretreatment of 2008 cells with 1 microM gemcitabine for 2 h before exposure to cisplatin for 7 h enhanced the accumulation of cisplatin IA and ICL by 50 and 40%, respectively (P < 0.05), above that induced by cisplatin alone. To investigate the possibility that the increased accumulation of cisplatin lesions was caused by inhibition of removal of cisplatin damage, 2008 cells were incubated with 200 microM cisplatin for 5 h in the presence and absence of gemcitabine and then a further 8 h in the absence of cisplatin. Only 57% IA were removed in the combination treated cells compared with 74% in cisplatin control cells. Similarly, repair of cisplatin ICL was inhibited in the gemcitabine-treated cells compared with the cells treated with cisplatin only (60 versus 72%). These findings demonstrate a direct inhibitory effect of gemcitabine on the repair of cisplatin IA and ICL and suggest a mechanistic basis for the cytotoxic synergy between the two drugs.


Bioengineered implantable scaffolds as a tool to study stromal-derived factors in metastatic cancer models.
Modeling the hematogenous spread of cancer cells to distant organs poses one of the greatest challenges in the study of human metastasis. Both tumor cell-intrinsic properties as well as interactions with reactive stromal cells contribute to this process, but identification of relevant stromal signals has been hampered by the lack of models allowing characterization of the metastatic niche. Here, we describe an implantable bioengineered scaffold, amenable to in vivo imaging, ex vivo manipulation, and serial transplantation for the continuous study of human metastasis in mice. Orthotopic or systemic inoculation of tagged human cancer cells into the mouse leads to the release of circulating tumor cells into the vasculature, which seed the scaffold, initiating a metastatic tumor focus. Mouse stromal cells can be readily recovered and profiled, revealing differential expression of cytokines, such as IL1β, from tumor-bearing versus unseeded scaffolds. Finally, this platform can be used to test the effect of drugs on suppressing initiation of metastatic lesions. This generalizable model to study cancer metastasis may thus identify key stromal-derived factors with important implications for basic and translational cancer research.


Breed-specific reference intervals for assessing thyroid function in seven dog breeds.
Thyroxine (T4), free T4 (FT4), and thyrotropin (TSH) concentrations were measured in serum from 693 healthy representatives from 7 dog breeds (Alaskan Malamute, Collie, English Setter, Golden Retriever, Keeshond, Samoyed, or Siberian Husky) to determine whether breed-specific reference intervals (RIs) are warranted. Veterinarians reviewed the health history, performed a physical examination, and approved laboratory data for the enrolled dogs. Many purebred dogs had T4 and FT4 concentrations that were at, or below, the lower limits previously determined for non-breed-specific RIs. Mean concentrations of T4, FT4, and TSH varied significantly among breeds. The range of mean concentration of T4 (19.7 nmol/L [1.53 µg/dL] in English Setters to 29.0 nmol/L [2.25 µg/dL] in Keeshonds) and FT4 (12.6 pmol/L [0.98 ng/dL] in English Setters to 20.2 pmol/L [1.57 ng/dL] in Samoyeds) was considerable. Median TSH values ranged from 6.10 mIU/L (0.07 ng/mL; Alaskan Malamute and Golden Retriever) to 17.6 mIU/L (0.26 ng/mL; Collie). Mean T4 and FT4 concentrations were higher in females. Increasing age was associated with decreasing T4 and FT4, and increasing TSH concentration. The substantial ranges across breeds of measures of central tendency (mean, median) for all hormones indicate that breed-specific RIs are warranted. RIs encompassing the central 95% of reference values for all breeds combined, and for individual breeds, were calculated using nonparametric (TSH) and robust (T4, FT4) methods. Use of breed-specific RIs in combination with careful attention to the potential for pre-analytical and analytical variability in test results will improve thyroid function assessment in these breeds.