Status, distribution and ecological risk of organochlorines (OCs) in the surface sediments from the Ravi River, Pakistan.
Organochlorines (OCs) including organochlorine pesticides (OCPs) and polychlorinated biphenyl (PCBs) were analyzed in surface sediments from the Ravi River, Punjab, Pakistan. Among the OCPs, hexachlorocyclohexane (HCHs), dichlorodiphenyltrichloroethane (DDTs) and chlordane (cis- and trans-) were most abundant and their concentrations were ranged from n.d to 16.0 ng g(-1), 1.5 to 58.5 ng g(-1) and n.d to 29.0 ng g(-1), respectively. Concentrations of Σ31PCBs ranged from 4.6 to 424.3 ng g(-1) with a mean value of 48.9 ng g(-1). In general, the concentrations of all studied organochlorines (OCs) in sediments collected from the sampling sites in the vicinity of industrial areas were higher than those from the agricultural areas. Compositional analyses of PCBs indicated that penta- and tetra-PCBs homologues were dominant which was consistent with the previous studies. Compared with other areas around the world, levels of OCs in sediments from the Ravi River were within the range, but were found to be much higher than previously reported from Pakistan. According to established sediment quality guidelines (SQGs), screening-level risk assessment of OCPs and PCBs suggested that they have a potential for adverse effects on benthic organisms.


Possible determination of the vector and reservoir of leishmaniasis in the Dominican Republic.
Two species of sand flies were collected by various methods from sites in the Dominican Republic. Lutzomyia cayennensis hispaniolae was the more common of the two. It was found in wooded habitats from sea level to an elevation of 442 m. This species was observed feeding on lizards (Anolis sp.) in the wild. In the laboratory, it fed only on lizards and only under lighted conditions. The other species, Lu. christophei was only found in the vicinity of seven leishmaniasis case sites. It readily fed on or probed rodents and humans. Although no naturally infected sand flies were collected, in the laboratory Lu. christophei was readily capable of transmitting the Dominican Leishmania parasite to uninfected BALB/c mice. We collected 167 specimens of three species of rodents and three Herpestes auropunctatus (mongoose) from the vicinity of two case sites. All four species are non-endemics introduced in post-Columbian times. Although we were unable to isolate parasites from any of these specimens, four of 44 Rattus rattus from one case site were seropositive for antibodies against Leishmania by indirect fluorescent antibody testing. This represents the first report of transmission of the Dominican Leishmania parasite by a sympatric species of sand fly and suggests that commensal rodents may play a role in the epidemiologic cycle.


A systematic review of fast food access studies.
The frequent consumption of energy-dense fast food is associated with increased body mass index. This systematic review aims to examine the methodology and current evidence on fast food access and its associations with outcomes. Six databases were searched using terms relating to fast food. Only peer-reviewed studies published in English during a 10-year period, with data collection and analysis regarding fast food access were included. Forty articles met the aforementioned criteria. Nearly half of the studies (n = 16) used their own set of features to define fast food. Studies predominantly examined the relationship between fast food access and socioeconomic factors (n = 21) and 76% indicated fast food restaurants were more prevalent in low-income areas compared with middle- to higher-income areas. Ten of 12 studies found fast food restaurants were more prevalent in areas with higher concentrations of ethnic minority groups in comparison with Caucasians. Six adult studies found higher body mass index was associated with living in areas with increased exposure to fast food; four studies, however, did not find associations. Further work is needed to understand if and how fast food access impacts dietary intake and health outcomes; and if fast food access has disparate socioeconomic, race/ethnicity and age associations.


Implanted Port Patency: Comparing Heparin and Normal Saline.
To maintain implanted port patency, clinicians can better determine the difference between the use of heparin and normal saline, including risk to patients, unnecessary cost to the healthcare system, and whether heparin is effective in maintaining port patency. The aim is to compare the effectiveness of saline to heparin for maintaining implanted port patency and to evaluate the cost differences. Data were collected from 56 patients enrolled in oncology clinical trials; 37 had paired data. Results showed that saline was as effective as heparin in maintaining implanted port patency. The difference in mean cost of saline versus heparin was statistically significant. Eliminating heparin when locking implanted ports did not increase catheter occlusion rates.


Diabetes and cancer: 5 years into the recent controversy.
Diabetes and cancer are common chronic disorders. The literature has long recognised that type 2 diabetes (T2D) is associated with an increased incident risk of several cancer types, independent of the mutual risk factor, obesity. However, in June 2009, four papers were published simultaneously in Diabetologia, the official journal of the European Association for the Study of Diabetes, raising questions of a link between diabetes therapies, notably the long-acting insulin analogue, glargine, and increased cancer risk. These papers awakened an unprecedented debate in the diabetes community, drawing in cancer experts and bringing together representatives from these two large, traditionally non-intersecting, biomedical communities. This Current Perspective summarises the events that followed the 'breaking news' from summer 2009: the pitfalls encountered; the increased mutual understanding between diabetes and cancer researchers; and the direction of current research. Much of the debate on the clinical impact of this controversy has been played out in the diabetes literature: here, we update the oncology readership.


Cell type-dependent differential activation of ERK by oncogenic KRAS in colon cancer and intestinal epithelium.
Oncogenic mutations in KRAS or BRAF are frequent in colorectal cancer and activate the ERK kinase. Here, we find graded ERK phosphorylation correlating with cell differentiation in patient-derived colorectal cancer organoids with and without KRAS mutations. Using reporters, single cell transcriptomics and mass cytometry, we observe cell type-specific phosphorylation of ERK in response to transgenic KRASG12V in mouse intestinal organoids, while transgenic BRAFV600E activates ERK in all cells. Quantitative network modelling from perturbation data reveals that activation of ERK is shaped by cell type-specific MEK to ERK feed forward and negative feedback signalling. We identify dual-specificity phosphatases as candidate modulators of ERK in the intestine. Furthermore, we find that oncogenic KRAS, together with β-Catenin, favours expansion of crypt cells with high ERK activity. Our experiments highlight key differences between oncogenic BRAF and KRAS in colorectal cancer and find unexpected heterogeneity in a signalling pathway with fundamental relevance for cancer therapy.


Neurocognitive performance during acute THC intoxication in heavy and occasional cannabis users.
Performance impairment during Delta(9)-tetrahydrocannabinol (THC) intoxication has been well described in occasional cannabis users. It is less clear whether tolerance develops to the impairing effects of THC in heavy users of cannabis. The aim of the present study was to assess neurocognitive performance during acute THC intoxication in occasional and heavy users. Twenty-four subjects (12 occasional cannabis users and 12 heavy cannabis users) participated in a double-blind, placebo-controlled, two-way mixed model design. Both groups received single doses of THC placebo and 500 microg/kg THC by smoking. Performance tests were conducted at regular intervals between 0 and 8 h after smoking, and included measures of perceptual motor control (critical tracking task), dual task processing (divided attention task), motor inhibition (stop signal task) and cognition (Tower of London). THC significantly impaired performance of occasional cannabis users on critical tracking, divided attention and the stop signal task. THC did not affect the performance of heavy cannabis users except in the stop signal task, i.e. stop reaction time increased, particularly at high THC concentrations. Group comparisons of overall performance in occasional and heavy users did not reveal any persistent performance differences due to residual THC in heavy users. These data indicate that cannabis use history strongly determines the behavioural response to single doses of THC.


Lipopolysaccharide-dependent interaction between PU.1 and c-Jun determines production of lipocalin-type prostaglandin D synthase and prostaglandin D2 in macrophages.
Previously, we reported that expression of lipocalin-prostaglandin D synthase (L-PGDS) is inducible in macrophages and protects from Pseudomonas pneumonia. Here, we investigated the mechanism by which L-PGDS gene expression is induced in macrophages. A promoter analysis of the murine L-PGDS promoter located a binding site of PU.1, a transcription factor essential for macrophage development and inflammatory gene expression. A chromatin immunoprecipitation assay showed that PU.1 bound to the cognate site in the endogenous L-PGDS promoter in response to LPS. Overexpression of PU.1, but not of PU.1(S148A), a mutant inert to casein kinase II (CKII) or NF-kappaB-inducing kinase (NIK), induced L-PGDS in RAW 264.7 cells. Conversely, siRNA silencing of PU.1 expression blunted productions of L-PGDS and prostaglandin D2 (PGD(2)). LPS treatment induced formation of the complex of PU.1 and cJun on the PU.1 site, but inactivation of cJun by treatment with JNK or p38 kinase inhibitor abolished the complex, and suppressed PU.1 transcriptional activity for L-PGDS gene expression. Together, these results show that PU.1, activated by CKII or NIK, cooperates with MAPK-activated cJun to maximally induce L-PGDS expression in macrophages following LPS treatment, and suggest that PU.1 participates in innate immunity through the production of L-PGDS and PGD(2).


C-arm flat detector computed tomography: the technique and its applications in interventional neuro-radiology.
Flat detector computed tomography (FDCT) is an imaging tool that generates three-dimensional (3-D) volumes from data obtained during C-arm rotation using CT-like reconstruction algorithms. The technique is relatively new and, at current levels of performance, lags behind conventional CT in terms of image quality. However, the advantage of its availability in the interventional room has prompted neuro-radiologists to identify clinical settings where its role is uniquely beneficial. We performed a search of the online literature databases to identify studies reporting experience with FDCT in interventional neuro-radiology. The studies were systematically reviewed and their findings grouped according to specific clinical situation addressed. FDCT images allow detection of procedural complications, evaluation of low-radiopacity stents and assessment of endosaccular coil packing in intra-cranial aneurysms. Additional roles are 3-D angiography that provides an accurate depiction of vessel morphology with low concentrations of radiographic contrast media and a potential for perfusion imaging due to its dynamic scanning capability. A single scan combining soft tissue and angiographic examinations reduces radiation dose and examination time. Ongoing developments in flat detector technology and reconstruction algorithms are expected to further enhance its performance and increase this range of applications. FDCT images provide useful information in neuro-interventional setting. If current research confirms its potential for assessing cerebral haemodynamics by perfusion scanning, the combination would redefine it as an invaluable tool for interventional neuro-radiology procedures. This facility and its existing capabilities of parenchymal and angiographic imaging would also extend its use to the triage of acute stroke patients.


Highly Flexible Protein-Peptide Docking Using CABS-Dock.
Protein-peptide molecular docking is a difficult modeling problem. It is even more challenging when significant conformational changes that may occur during the binding process need to be predicted. In this chapter, we demonstrate the capabilities and features of the CABS-dock server for flexible protein-peptide docking. CABS-dock allows highly efficient modeling of full peptide flexibility and significant flexibility of a protein receptor. During CABS-dock docking, the peptide folding and binding process is explicitly simulated and no information about the peptide binding site or its structure is used. This chapter presents a successful CABS-dock use for docking a potentially therapeutic peptide to a protein target. Moreover, simulation contact maps, a new CABS-dock feature, are described and applied to the docking test case. Finally, a tutorial for running CABS-dock from the command line or command line scripts is provided. The CABS-dock web server is available from http://biocomp.chem.uw.edu.pl/CABSdock/ .


Qualified Biolayer Interferometry Avidity Measurements Distinguish the Heterogeneity of Antibody Interactions with Plasmodium falciparum Circumsporozoite Protein Antigens.
Ab avidity is a measure of the overall strength of Ab-Ag interactions and hence is important for understanding the functional efficiency of Abs. In vaccine evaluations, Ab avidity measurements can provide insights into immune correlates of protection and generate hypotheses regarding mechanisms of protection to improve vaccine design to achieve higher levels of efficacy. The commonly used Ab avidity assays require the use of chaotropic reagents to measure avidity index. In this study, using real-time detection of Ab-Ag binding by biolayer interferometry (BLI) technique, we have developed a qualified assay for measuring avidity of vaccine-induced Abs specific for Plasmodium falciparum circumsporozoite protein (CSP) Ags. Human mAb derived from plasmablasts of recipients of RTS,S/AS01 (RTS,S), the most advanced malaria vaccine candidate, were used in the assay development to measure Ag-specific binding responses and rate constants of association and dissociation. The optimized BLI binding assay demonstrated 1) good precision (percentage of coefficient of variation <20), 2) high specificity, 3) a lower limit of detection and quantitation in the 0.3-3.3 nM range, and 4) a range of linearity up to 50-100 nM for the CSP Ags tested. Analysis of polyclonal sera of malaria vaccinees demonstrated the suitability of this method to distinguish among vaccinees and rank Ab responses by avidity. These results demonstrate that precise, specific, and sensitive BLI measurements of Ab avidity in polyclonal sera from malaria vaccinees can map Ab response heterogeneity and potentially help to determine the role of Ab avidity as an immune correlate of protection for vaccines.


Detrimental influence on performance of high temperature incubation in a tropical reptile: is cooler better in the tropics?
Global temperatures have risen over the last century, and are forecast to continue rising. Ectotherms may be particularly sensitive to changes in thermal regimes, and tropical ectotherms are more likely than temperate species to be influenced by changes in environmental temperature, because they may have evolved narrow thermal tolerances. Keelback snakes (Tropidonophis mairii) are tropical, oviparous reptiles. To quantify the effects of temperature on the morphology and physiology of hatchling keelbacks, clutches laid by wild-caught females were split and incubated at three temperatures, reflecting the average minimum, overall average and average maximum temperatures recorded at our study site. Upon hatching, the performance of neonates was examined at all three incubation temperatures in a randomized order over consecutive days. Hatchlings from the 'hot' treatment had slower burst swim speeds and swam fewer laps than hatchlings from the cooler incubation temperatures in all three test temperatures, indicating a low thermal optimum for incubation of this tropical species. There were no significant interactions between test temperature and incubation temperature across performance variables, suggesting phenotypic differences caused by incubation temperature did not acclimate this species to post-hatching conditions. Thus, keelback embryos appear evolutionarily adapted to development at cooler temperatures (relative to what is available in their habitat). The considerable reduction in hatchling viability and performance associated with a 3.5 °C increase in incubation temperature, suggests climate change may have significant population-level effects on this species. However, the offspring of three mothers exposed to the hottest incubation temperature were apparently resilient to high temperature, suggesting that this species may respond to selection imposed by thermal regime.


Modification by N-acetyltransferase 1 genotype on the association between dietary heterocyclic amines and colon cancer in a multiethnic study.
Colorectal cancer incidence is greater among African Americans, compared to whites in the U.S., and may be due in part to differences in diet, genetic variation at metabolic loci, and/or the joint effect of diet and genetic susceptibility. We examined whether our previously reported associations between meat-derived heterocyclic amine (HCA) intake and colon cancer were modified by N-acetyltransferase 1 (NAT1) or 2 (NAT2) genotypes and whether there were differences by race. In a population-based, case-control study of colon cancer, exposure to HCAs was assessed using a food-frequency questionnaire with a meat-cooking and doneness module, among African Americans (217 cases and 315 controls) and whites (290 cases and 534 controls). There was no association with NAT1*10 versus NAT1-non*10 genotypes for colon cancer. Among whites, there was a positive association for NAT2-"rapid/intermediate" genotype [odds ratio (OR)=1.4; 95% confidence interval (CI)=1.0, 1.8], compared to the NAT2-"slow" that was not observed among African Americans. Colon cancer associations with HCA intake were modified by NAT1, but not NAT2, regardless of race. However, the "at-risk" NAT1 genotype differed by race. For example, among African Americans, the positive association with 2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine (PhIP) was confined to those with NAT1*10 genotype (OR=1.8; 95% CI=1.0, 3.3; P for interaction=0.02, comparing highest to lowest intake), but among whites, an association with 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) was confined to those with NAT1-non*10 genotype (OR=1.9; 95% CI=1.1, 3.1; P for interaction=0.03). Our data indicate modification by NAT1 for HCA and colon cancer associations, regardless of race. Although the at-risk NAT1 genotype differs by race, the magnitude of the individual HCA-related associations in both race groups are similar. Therefore, our data do not support the hypothesis that NAT1 by HCA interactions contribute to differences in colorectal cancer incidence between African Americans and whites.


Ethnic differences in maternal dietary patterns are largely explained by socio-economic score and integration score: a population-based study.
The impact of socio-economic position and integration level on the observed ethnic differences in dietary habits has received little attention. To identify and describe dietary patterns in a multi-ethnic population of pregnant women, to explore ethnic differences in odds ratio (OR) for belonging to a dietary pattern, when adjusted for socio-economic status and integration level and to examine whether the dietary patterns were reflected in levels of biomarkers related to obesity and hyperglycaemia. This cross-sectional study was a part of the STORK Groruddalen study. In total, 757 pregnant women, of whom 59% were of a non-Western origin, completed a food frequency questionnaire in gestational week 28±2. Dietary patterns were extracted through cluster analysis using Ward's method. Four robust clusters were identified where cluster 4 was considered the healthier dietary pattern and cluster 1 the least healthy. All non-European women as compared to Europeans had higher OR for belonging to the unhealthier dietary patterns 1-3 vs. cluster 4. Women from the Middle East and Africa had the highest OR, 21.5 (95% CI 10.6-43.7), of falling into cluster 1 vs. 4 as compared to Europeans. The ORs decreased substantially after adjusting for socio-economic score and integration score. A non-European ethnic origin, low socio-economic and integration scores, conduced higher OR for belonging to clusters 1, 2, and 3 as compared to cluster 4. Significant differences in fasting and 2-h glucose, fasting insulin, glycosylated haemoglobin (HbA1c), insulin resistance (HOMA-IR), and total cholesterol were observed across the dietary patterns. After adjusting for ethnicity, differences in fasting insulin (p=0.015) and HOMA-IR (p=0.040) across clusters remained significant, despite low power. The results indicate that socio-economic and integration level may explain a large proportion of the ethnic differences in dietary patterns.


Genome-wide expression profiling of lymphoblastoid cell lines distinguishes different forms of autism and reveals shared pathways.
Autism is a heterogeneous condition that is likely to result from the combined effects of multiple genetic factors interacting with environmental factors. Given its complexity, the study of autism associated with Mendelian single gene disorders or known chromosomal etiologies provides an important perspective. We used microarray analysis to compare the mRNA expression profile in lymphoblastoid cells from males with autism due to a fragile X mutation (FMR1-FM), or a 15q11-q13 duplication (dup(15q)), and non-autistic controls. Gene expression profiles clearly distinguished autism from controls and separated individuals with autism based on their genetic etiology. We identified 68 genes that were dysregulated in common between autism with FMR1-FM and dup(15q). We also identified a potential molecular link between FMR1-FM and dup(15q), the cytoplasmic FMR1 interacting protein 1 (CYFIP1), which was up-regulated in dup(15q) patients. We were able to confirm this link in vitro by showing common regulation of two other dysregulated genes, JAKMIP1 and GPR155, downstream of FMR1 or CYFIP1. We also confirmed the reduction of the Jakmip1 protein in Fmr1 knock-out mice, demonstrating in vivo relevance. Finally, we showed independent confirmation of roles for JAKMIP1 and GPR155 in autism spectrum disorders (ASDs) by showing their differential expression in male sib pairs discordant for idiopathic ASD. These results provide evidence that blood derived lymphoblastoid cells gene expression is likely to be useful for identifying etiological subsets of autism and exploring its pathophysiology.


The subscapular and subcoracoid bursae: descriptive and functional anatomy.
The subscapularis and subcoracoid bursae, as well as the subscapularis muscle, were studied in 42 fresh cadaveric shoulders. Fibrous bands were found in the medial part of the muscle; they were intercalated with the distal tendon-like bands. The superior distal fibrous band was always found to be thicker than the others. Strong fibrous attachments of the subscapularis muscle were found along the lateral border of the scapula as well as along the medial third of the bony crests in the subscapular fossa. Between the crests, the muscle bundles were directly attached to the bone. In the lateral part of the scapula, the subscapularis muscle had only weak connective links with the bone. The subscapularis bursa was found in all cases as a pouch strongly attached to the scapular neck and the adjacent part of the joint capsule. The top of the bursa was linked to the coracoid process by a fibrous attachment called the suspensory ligament. The subcoracoid bursa had only weak connective links with the coracoid process. In 28% of cases, the subscapularis and subcoracoid bursae merged into a unique wide bursa. The subscapular bursa had two types of links with the muscle: (1). weak connective links with the superficial muscle bundles and (2). at least 2 fibrous arcs joining the lateral tendon-like bands, the bursa, and bony crests of the subscapularis fossa. These arcs allow the bursa to follow the course of the muscle exactly. During movements of the glenohumeral joint, the subscapularis muscle sustains huge changes of orientation, particularly the upper part of the muscle that coils around the coracoid process. The strong superior fibrous band enables the muscle to maintain contact with the coracoid process. It is the function of the subscapularis and subcoracoid bursae to manage the friction of the superficial fibers against the scapular neck, the humeral head, and the coracoid process.


Selection, generalized transmission and the evolution of modifier genes. I. The reduction principle.
Modifier gene models are used to explore the evolution of features of organisms, such as the genetic system, that are not directly involved in the determination of fitness. Recent work has shown that a general "reduction principle" holds in models of selectively neutral modifiers of recombination, mutation, and migration. Here we present a framework for models of modifier genes that shows these reduction results to be part of a more general theory, for which recombination and mutation are special cases. The deterministic forces that affect the genetic composition of a population can be partitioned into two categories: selection and transmission. Selection includes differential viabilities, fertilities, and mating success. Imperfect transmission occurs as a result of such phenomena as recombination, mutation and migration, meiosis, gene conversion, and meiotic drive. Selectively neutral modifier genes affect transmission, and a neutral modifier gene can evolve only by generating association with selected genes whose transmission it affects. We show that, in randomly mating populations at equilibrium, imperfect transmission of selected genes allows a variance in their marginal fitnesses to be maintained. This variance in the marginal fitnesses of selected genes is what drives the evolution of neutral modifier genes. Populations with a variance in marginal fitnesses at equilibrium are always subject to invasion by modifier genes that bring about perfect transmission of the selected genes. It is also found, within certain constraints, that for modifier genes producing what we call "linear variation" in the transmission processes, a new modifier allele can invade a population at equilibrium if it reduces the level of imperfect transmission acting on the selected genes, and will be expelled if it increases the level of imperfect transmission. Moreover, the strength of the induced selection on the modifier gene is shown to range up to the order of the departure of the genetic system from perfect transmission.


Accelerating the switchgrass (Panicum virgatum L.) breeding cycle using genomic selection approaches.
Switchgrass (Panicum virgatum L.) is a perennial grass undergoing development as a biofuel feedstock. One of the most important factors hindering breeding efforts in this species is the need for accurate measurement of biomass yield on a per-hectare basis. Genomic selection on simple-to-measure traits that approximate biomass yield has the potential to significantly speed up the breeding cycle. Recent advances in switchgrass genomic and phenotypic resources are now making it possible to evaluate the potential of genomic selection of such traits. We leveraged these resources to study the ability of three widely-used genomic selection models to predict phenotypic values of morphological and biomass quality traits in an association panel consisting of predominantly northern adapted upland germplasm. High prediction accuracies were obtained for most of the traits, with standability having the highest ten-fold cross validation prediction accuracy (0.52). Moreover, the morphological traits generally had higher prediction accuracies than the biomass quality traits. Nevertheless, our results suggest that the quality of current genomic and phenotypic resources available for switchgrass is sufficiently high for genomic selection to significantly impact breeding efforts for biomass yield.


Factors associated with poor sleep quality in patients with multiple sclerosis differ by disease duration.
Sleep disturbance is a common symptom of multiple sclerosis (MS) and knowledge about factors that contribute to poor sleep quality is scarce. The aim was to explore the differences in the prevalence and determinants of poor sleep quality in a sample of patients with MS with disease duration ≤ 5 years and >5 years. We collected data from 152 consecutive patients with MS; 66 patients (78% women, averaged 37.35 ± 10.1 years) were in the group with disease duration ≤ 5 years and 86 patients (73.3% women, averaged 42.10 ± 9.4 years) in the group with disease duration >5 years. Patients filled out the Pittsburgh Sleep Quality Index, the Hospital Anxiety and Depression Scale, the Multidimensional Fatigue Inventory, one item of the Incapacity Status Scale regarding bladder problems and one item of the Short Form-36 regarding pain. Multiple linear regression was used to analyze the relationship between the study variables. The prevalence of poor sleep is significantly higher in patients with longer disease duration (34.8 vs. 51.2%). Anxiety, reduced motivation and mental fatigue (all p < 0.05) were associated with poor sleep quality in patients with disease duration ≤5 years, whereas pain (p < 0.01), depression and mental fatigue (both p < 0.05) were in patients with disease duration >5 years. Sleep problems are present in patients with MS with both short and long disease duration, but these problems are associated with different factors. These should be recognized and managed in addition to the treatment of sleep disorders.


Biochemical comparison of osteoarthritic knees with and without effusion.
Several symptom-relieving interventions have been shown to be efficacious among osteoarthritis (OA) patients with knee effusion; however, not every symptomatic knee OA patient has clinical effusion. Results may be over-generalized since it is unclear if effused knees represent a unique pathological condition or subset compared to knees without effusion. The primary purpose of this study was to determine if biochemical differences existed between OA knees with and without effusion. The present cross-sectional study consisted of 22 volunteers (11 with knee effusion, 11 without knee effusion) with confirmed late-stage radiographic knee OA (Kellgren-Lawrence score ≥ 3). Synovial fluid samples were collected and analyzed using a custom multiplex enzyme-linked immunosorbent assay to determine eight specific biomarker concentrations (e.g., catabolic, anabolic). Matrix metalloproteinase (MMP)-3, tissue inhibitor of MMPs (TIMP)-1, TIMP-2, and interleukin-10 were significantly higher in the knees with effusion than in the knees without effusion. The biochemical differences that existed between knees with and without effusion provide support that OA subsets may exist, characterized by distinct biochemical characteristics and clinical findings (e.g., effusion).


Site preferences in hetero-metallic [Fe9-xNix] clusters: a combined crystallographic, spectroscopic and theoretical analysis.
The reaction of mixtures of Fe(O2CMe)2·2H2O and Ni(O2CMe)2·4H2O of various compositions with di-2-pyridyl ketone (py2CO, dpk) in MeCN under an inert atmosphere afforded a family of hetero-metallic enneanuclear clusters with general formula [Fe9-xNix(μ4-OH)2(O2CMe)8(py2CO2)4] (2, x = 1.00; 3: x = 6.02; 4, x = 7.46; 5, x = 7.81). Clusters 2-5 are isomorphous to the homo-metallic [Fe9] cluster (1) previously reported by some of us, and also isostructural to the known homo-metallic [Ni9] cluster. All four clusters contain a central MII ion in an unusual 8-coordinate site and eight peripheral MII ions in distorted octahedral environments. The distribution of FeII and NiII ions over these two distinct coordination sites in 2-5 can be established through a combination of X-ray fluorescence and Mössbauer spectroscopies, which show that FeII preferentially occupies the unique 8-coordinate metal site while NiII accumulates in the octahedral holes. Density functional theory indicates that the distribution of ions across the two sites arises not from any intrinsic preference of the FeII ions for the 8-coordinate sites, but rather because of the large ligand field stabilization energy available to NiII in octahedral coordination.


Childhood Family Structure and Intergenerational Income Mobility in the United States.
The declining prevalence of two-parent families helped increase income inequality over recent decades. Does family structure also condition how economic (dis)advantages pass from parents to children? If so, shifts in the organization of family life may contribute to enduring inequality between groups defined by childhood family structure. Using National Longitudinal Survey of Youth data, I combine parametric and nonparametric methods to reveal how family structure moderates intergenerational income mobility in the United States. I find that individuals raised outside stable two-parent homes are much more mobile than individuals from stable two-parent families. Mobility increases with the number of family transitions but does not vary with children's time spent coresiding with both parents or stepparents conditional on a transition. However, this mobility indicates insecurity, not opportunity. Difficulties maintaining middle-class incomes create downward mobility among people raised outside stable two-parent homes. Regardless of parental income, these people are relatively likely to become low-income adults, reflecting a new form of perverse equality. People raised outside stable two-parent families are also less likely to become high-income adults than people from stable two-parent homes. Mobility differences account for about one-quarter of family-structure inequalities in income at the bottom of the income distribution and more than one-third of these inequalities at the top.


Kinetic mechanism of DNA polymerase I (Klenow).
The minimal kinetic scheme for DNA polymerization catalyzed by the Klenow fragment of DNA polymerase I (KF) from Escherichia coli has been determined with short DNA oligomers of defined sequence. A key feature of this scheme is a minimal two-step sequence that interconverts the ternary KF.DNAn.dNTP and KF.DNAn+1.PPi complexes. The rate is not limited by the actual polymerization but by a separate step, possibly important in ensuring fidelity [Mizrahi, V., Henrie, R. N., Marlier, J. F., Johnson, K. A., & Benkovic, S. J. (1985) Biochemistry 24, 4010-4018]. Evidence for this sequence is supplied by the observation of biphasic kinetics in single-turnover pyrophosphorolysis experiments (the microscopic reverse of polymerization). Data analysis then provides an estimate of the internal equilibrium constant. The dissociations of DNA, dNTP, and PPi from the various binary and ternary complexes were measured by partitioning (isotope-trapping) experiments. The rate constant for DNA dissociation from KF is sequence dependent and is rate limiting during nonprocessive DNA synthesis. The combination of single-turnover (both directions) and isotope-trapping experiments provides sufficient information to permit a quantitative evaluation of the kinetic scheme for specific DNA sequences.


Atopic characteristics of children with recurrent wheezing at high risk for the development of childhood asthma.
Few studies have characterized the atopic profile of toddler-aged children with recurrent wheezing at high risk of the development of persistent asthma. Objective We sought to determine the atopic profile of toddler-aged children with frequent wheeze at high risk for the development of persistent asthma who either had a parental history of asthma, a personal history of atopic dermatitis, or both. Participants enrolled in the Prevention of Early Asthma in Kids study (n = 285) on the basis of a modified Asthma Predictive Index were characterized on the basis of allergy and asthma questionnaire responses and allergy skin puncture test results. The majority of the children (60.7%, n = 148) were sensitized to either food or aeroallergens. Male children were significantly more likely to be sensitized to aeroallergens ( P = .03) and to have a blood eosinophil level of 4% or greater ( P = .03) and a total serum IgE level of greater than 100 IU/mL ( P = .0004). Additionally, eosinophilia and total serum IgE level had the strongest correlation with aeroallergen sensitization. The high prevalence of aeroallergen sensitization in this high-risk cohort suggests that aeroallergens might have an important role in the early development of asthma. As such, the Prevention of Early Asthma in Kids cohort appears to be an appropriate cohort in which to test whether early intervention with an inhaled corticosteroid can significantly attenuate, or perhaps even prevent, the allergic march from the initial stages of allergic sensitization to the subsequent development of asthma in toddlers with episodic wheezing.


Immune escape of the human facultative pathogenic yeast Candida albicans: the many faces of the Candida Pra1 protein.
Infectious diseases caused by human pathogenic fungi represent a major and global health problem. Based on the limited efficacy of existing drugs and the increasing resistance to antifungal compounds, new strategies are urgently needed to fight such fungal infections. The medically important pathogen Candida albicans can exist as an opportunistic yeast, but can also cause severe diseases, septicaemia, and death. In order to establish new strategies to fight Candida infections and to interfere with survival of the pathogen, it is highly relevant to understand the molecular and immunology interactions between the pathogen C. albicans and the human host. This immune cross talk has moved into the focus of infectious disease research. In this review, we summarize the diverse and multiple levels of the immune cross talk between the fungal pathogen C. albicans and the human host. In particular, we define how one single fungal protein Pra1 (pH-regulated antigen 1) interferes and controls host immune attack at multiple levels and thus contributes to fungal immune escape. Candida Pra1 represents a promising candidate for immune interference.


The effects of S-nitrosocaptopril on canine coronary circulation.
Nitroglycerin (NTG) has been used for more than a century for the relief of angina pectoris and is regarded as an extrinsic donor of nitric oxide (NO). Captopril, an angiotensin-converting enzyme inhibitor (ACEI), is reported to have beneficial effects on survival in acute myocardial infarction. In this study, a hybrid compound of NO and ACEI, S-nitrosocaptopril (S-NO-Cap), was synthesized, which was characterized by its molecular structure. Its coronary effects in chronically instrumented dogs were compared with those of NTG and captopril. S-NO-Cap (50 micrograms/kg i.v.) increased epicardial coronary diameter (CoD) by a maximum of 5.2% for more than 20 min. NTG (15 micrograms/kg i.v.) increased CoD by a maximum of 4.6% for 10 min, i.e., the effect was equipotent with that of S-NO-Cap (50 micrograms/kg i.v.). However, S-NO-Cap produced its maximal CoD increase more slowly than NTG did. Both S-NO-Cap and NTG transiently increased coronary blood flow. However, NTG had a more potent effect than S-NO-Cap (28.4 vs. 40.8 ml/min, respectively). Captopril, on the other hand, had almost no effect on either CoD or coronary blood flow. Thus, the vasodilatory action of S-NO-Cap more closely resembled that of NTG than that of captopril. Therefore, S-NO-Cap may dilate coronary arteries by virtue of its NO moiety rather than by its ACEI properties. These findings indicate that S-NO-Cap is a potential antianginal drug.


Clinical acidosis in a Gippsland dairy herd.
To report on spontaneous clinical and subclinical acidosis in a large dairy herd, to evaluate the diets and feeding strategies involved, and to report on measures of rumen function in the cows affected. A Gippsland dairy herd was sampled as part of a wider randomised cross-sectional study that examined the prevalence, risk factors for, and effects of, acidosis on rumen function of dairy cattle. Three herds on the farm were involved in the study: the transition herd (cows 3 weeks prior to calving), the very fresh lactating herd (1 < days in milk < 10, herd 1) and the fresh lactating herd (10 < days in milk < 120, herd 2) including a small lame herd fed separately. The transition cows were fed 2 kg dry matter triticale per cow per day and hay with an estimated total dry matter intake of 4.8 kg per cow per day. The lactating cow diet included 6.75 kg dry matter triticale per cow per day with total concentrate fed being 8.1 kg dry matter per cow per day in the milking parlour. Silage, lucerne cubes, hay and pasture (herd 2 only) was also fed to the lactating cows with the estimated total dry matter intake for cows in herds 1 and 2 being 13.7 kg and 20.8 kg per cow per day respectively. Three primiparous and five multiparous cows in early lactation (< 100 days in milk) were randomly selected from each of two lactating herds: herds 1 and 2. Rumen fluid was sampled from each cow by both rumenocentesis and stomach tube. The rumenocentesis samples were tested for pH at the time of sampling. Stomach tube samples were frozen and subsequently tested for volatile fatty acid, ammonia, and D- and L-lactate concentration. In the very fresh herd, there was a high prevalence of severe lameness and scouring, coupled with a mean rumen pH 5.67, low mean total volatile fatty acid concentration 71.0 mM and high mean concentrations of L- and D-lactate, (7.71 mM and 7.31 mM), respectively. Cows in the fresh herd had moderate signs of scouring and lameness. A lame herd comprising approximately 50 cows separated from the fresh herd was also present on the farm. The mean rumen pH of the fresh herd was 5.74 and mean rumen concentrations of volatile fatty acids, ammonia, L- and D-lactate were within ranges considered normal. The transition diet failed to supply sufficient energy and protein for maintenance of cows of this live weight in late gestation. The diet fed to the very fresh herd was low in effective fibre and contained an excessive content of non-structural carbohydrate in the form of processed, rapidly fermentable grain. The sudden change from the transition diet to the diet fed to the very fresh herd probably also precipitated the outbreak of acidosis. This case report provides unique detail, including information on diets and rumen parameters, of an outbreak of acidosis in a pasture-fed herd.


Reproducibility of data from a hand-held digital pulp tester used on teeth and oral soft tissue.
Controlled studies of the reproducibility of data from electronic pulp testing instruments are limited and contain few statistical analyses. The reproducibility of these readings is important if the instrument is to be used for determining differences in sensitivity. Twenty human subjects (16 male) were used in this study. One incisor, one premolar, one molar tooth with small or no restorations, and two gingival soft tissue positions from each upper and lower arch of each subject were stimulated with the Analytic Technology vitality scanner. This procedure was repeated twice with a 5-minute rest between each trial, for a total of three trials. Each subject was then seen again after a period of at least 3 days, at which time the trials were repeated. The collected data were grouped by trial, tooth position, and day. Paired t test analysis of both the absolute difference between any two trials on the same day and the average of the absolute differences between corresponding trials on days 1 and 2 showed no statistically significant differences (p greater than 0.05). Accommodation to the stimulus was evaluated by examining differences in the mean values between the three same-day trials. The Analytic Technology vitality scanner was found to be reproducible both for consecutive same-day trials and for corresponding trials on different days. No same-day trends in meter readings were noted.


The implications of a tracheal bronchus on one-lung ventilation and fibreoptic bronchoscopy in a patient undergoing thoracic surgery: a case report.
Due to its anatomical complexity, a tracheal bronchus has important clinical implications for one-lung ventilation (OLV). We present a case of successful OLV in a patient with a high a type I (i.e., high take-off) tracheal bronchus. This anomaly presented unusual fibreoptic bronchoscopic (FOB) views that were difficult to discern from the normal carinal bifurcation. A 35-yr-old male presented for posterior basal segmentectomy of the left lower lobe under video-assisted thoracoscopy. The preoperative chest radiography was reported as normal, but a computed tomography scan of the chest revealed a right upper lobe tracheal bronchus. The inlet of the tracheal bronchus was located high above the carina, and the distal trachea had significant narrowing. Because the main trachea was divided into a tracheal bronchus and a distal trachea with similar diameters and with an acute angle of divergence, FOB views of the tracheal bronchus take-off appeared similar to the normal carinal bifurcation. Moreover, the actual carina had an atypical appearance with the main bronchi shifted laterally and a blunted carinal ridge. As a result of this atypical tracheobronchial anatomy, we used an Arndt endobronchial blocker system instead of a double-lumen tube (DLT) for right-sided OLV. One-lung ventilation was satisfactory throughout the uncomplicated operation. Careful preoperative assessment of tracheobronchial anatomy is imperative in order to choose an appropriate method of OLV and prevent potential complications. In a type I tracheal bronchus with a narrowed distal trachea, a bronchial blocker may have advantages over the conventional DLT in achieving OLV.


Gene cluster of Rhodothermus marinus high-potential iron-sulfur Protein: oxygen oxidoreductase, a caa(3)-type oxidase belonging to the superfamily of heme-copper oxidases.
The respiratory chain of the thermohalophilic bacterium Rhodothermus marinus contains an oxygen reductase, which uses HiPIP (high potential iron-sulfur protein) as an electron donor. The structural genes encoding the four subunits of this HiPIP:oxygen oxidoreductase were cloned and sequenced. The genes for subunits II, I, III, and IV (named rcoxA to rcoxD) are found in this order and seemed to be organized in an operon of at least five genes with a terminator structure a few nucleotides downstream of rcoxD. Examination of the amino acid sequence of the Rcox subunits shows that the subunits of the R. marinus enzyme have homology to the corresponding subunits of oxidases belonging to the superfamily of heme-copper oxidases. RcoxB has the conserved histidines involved in binding the binuclear center and the low-spin heme. All of the residues proposed to be involved in proton transfer channels are conserved, with the exception of the key glutamate residue of the D-channel (E(278), Paracoccus denitrificans numbering). Analysis of the homology-derived structural model of subunit I shows that the phenol group of a tyrosine (Y) residue and the hydroxyl group of the following serine (S) may functionally substitute the glutamate carboxyl in proton transfer. RcoxA has an additional sequence for heme C binding, after the Cu(A) domain, that is characteristic of caa(3) oxidases belonging to the superfamily. Homology modeling of the structure of this cytochrome domain of subunit II shows no marked electrostatic character, especially around the heme edge region, suggesting that the interaction with a redox partner is not of an electrostatic nature. This observation is analyzed in relation to the electron donor for this caa(3) oxidase, the HiPIP. In conclusion, it is shown that an oxidase, which uses an iron-sulfur protein as an electron donor, is structurally related to the caa(3) class of heme-copper cytochrome c oxidases. The data are discussed in the framework of the evolution of oxidases within the superfamily of heme-copper oxidases.


Audiovisual asynchrony detection in human speech.
Combining information from the visual and auditory senses can greatly enhance intelligibility of natural speech. Integration of audiovisual speech signals is robust even when temporal offsets are present between the component signals. In the present study, we characterized the temporal integration window for speech and nonspeech stimuli with similar spectrotemporal structure to investigate to what extent humans have adapted to the specific characteristics of natural audiovisual speech. We manipulated spectrotemporal structure of the auditory signal, stimulus length, and task context. Results indicate that the temporal integration window is narrower and more asymmetric for speech than for nonspeech signals. When perceiving audiovisual speech, subjects tolerate visual leading asynchronies, but are nevertheless very sensitive to auditory leading asynchronies that are less likely to occur in natural speech. Thus, speech perception may be fine-tuned to the natural statistics of audiovisual speech, where facial movements always occur before acoustic speech articulation.


Adverse testicular effects of Botox® in mature rats.
Botox® injections are taking a consistently increasing place in urology. Intracremasteric injections, particularly, have been applied for cryptorchidism and painful testicular spasms. Studies outlining their safety for this use are, however, scanty. Thus, the present study aimed at evaluating possible testicular toxicity of Botox® injections and their effect on male fertility. Mature rats were given intracremasteric Botox® injections (10, 20 and 40 U/kg) three times in a two-week interval. Changes in body and testes weights were examined and gonadosomatic index compared to control group. Semen quality, sperm parameters, fructose, protein, cholesterol and triglycerides contents were assessed. Effects on normal testicular function were investigated by measuring testosterone levels and changes in enzyme activities (lactate dehydrogenase-X and acid phosphatase). To draw a complete picture, changes in oxidative and inflammatory states were examined, in addition to the extent of connective tissue deposition between seminiferous tubules. In an attempt to have more accurate information about possible spermatotoxic effects of Botox®, flowcytometric analysis and histopathological examination were carried out. Botox®-injected rats showed altered testicular physiology and function. Seminiferous tubules were separated by dense fibers, especially with the highest dose. Flowcytometric analysis showed a decrease in mature sperms and histopathology confirmed the findings. The oxidative state was, however, comparable to control group. This study is the first to show that intracremasteric injections of Botox® induce adverse testicular effects evidenced by inhibited spermatogenesis and initiation of histopathological changes. In conclusion, decreased fertility may be a serious problem Botox® injections could cause.


Polysaccharide from Pleurotus nebrodensis induces apoptosis via a mitochondrial pathway in HepG2 cells.
A novel alkali extractable polysaccharide (designated as PNA-2) was purified from Pleurotus nebrodensis and the effects of purified PNA-2 on the proliferation and apoptosis of human hepatic cancer cells (HepG2) were investigated in this study. The results of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated that PNA-2 inhibited the proliferation of HepG2 cells by apoptosis induction, which was also characterized using scanning electron microscopy (SEM). Moreover, the expression of apoptosis-associated mRNA, proteins and the cell-cycle arrest at the G0/G1 phase was determined using RT-qPCR, Western blot and flow cytometry, respectively. A notable inhibition of the migration rate of PNA-2-treated HepG2 cells was observed using a cell scratch assay. DNA damage was observed using a comet assay and AO/EB staining in HepG2 cells, which were exposed to PNA-2. Induction of the mitochondria-mediated intrinsic apoptotic pathway by PNA-2 was indicated by the loss of mitochondrial membrane potential (ΔΨm), Bcl-2 dysregulation and cytochrome c release. All the results suggested that the mitochondria-mediated intrinsic apoptotic pathway could be involved in PNA-2-mediated apoptosis of human liver carcinoma cells HepG2. Finally, the results indicated that PNA-2 significantly suppressed tumor growth in HepG2 tumor-bearing mice, indicating that PNA-2 may be developed as a candidate drug or functional food factor to prevent or treat liver cancer.


Chirped-mirror dispersion-compensated femtosecond optical parametric oscillator.
We describe the operating characteristics of a femtosecond optical parametric oscillator employing chirped mirrors for intracavity group-velocity dispersion compensation. Pumped by 760 mW of power from a self-mode-locked Ti:sapphire laser, this device provides 100-fs near-transform-limited pulses continuously tunable from 1.18 to 1.32 microm with an average power of 100-180 mW. The limitations of the present setup and strategies for further pulse shortening are discussed.


A major external source of cholinergic innervation of the striatum and nucleus accumbens originates in the brainstem.
Cholinergic transmission in the striatal complex is critical for the modulation of the activity of local microcircuits and dopamine release. Release of acetylcholine has been considered to originate exclusively from a subtype of striatal interneuron that provides widespread innervation of the striatum. Cholinergic neurons of the pedunculopontine (PPN) and laterodorsal tegmental (LDT) nuclei indirectly influence the activity of the dorsal striatum and nucleus accumbens through their innervation of dopamine and thalamic neurons, which in turn converge at the same striatal levels. Here we show that cholinergic neurons in the brainstem also provide a direct innervation of the striatal complex. By the expression of fluorescent proteins in choline acetyltransferase (ChAT)::Cre(+) transgenic rats, we selectively labeled cholinergic neurons in the rostral PPN, caudal PPN, and LDT. We show that cholinergic neurons topographically innervate wide areas of the striatal complex: rostral PPN preferentially innervates the dorsolateral striatum, and LDT preferentially innervates the medial striatum and nucleus accumbens core in which they principally form asymmetric synapses. Retrograde labeling combined with immunohistochemistry in wild-type rats confirmed the topography and cholinergic nature of the projection. Furthermore, transynaptic gene activation and conventional double retrograde labeling suggest that LDT neurons that innervate the nucleus accumbens also send collaterals to the thalamus and the dopaminergic midbrain, thus providing both direct and indirect projections, to the striatal complex. The differential activity of cholinergic interneurons and cholinergic neurons of the brainstem during reward-related paradigms suggest that the two systems play different but complementary roles in the processing of information in the striatum.


The real response of bone to exercise.
This review presents findings made in studies of large mammalian bones, especially from racehorse training experiments (2-8 years old, third metacarpal, tarsal) and human autopsy orthopaedic femoral implant retrievals and other human biopsy and autopsy cases. Samples were cleaned to analyse mineralized matrix in three dimensions, or poly methyl-methacrylate embedded and micromilled to delete topography and study the superficial c. 0.5-microm two-dimensional section using quantitative backscattered electron imaging. With experimental implant studies in rabbits, observations were also made in vivo using confocal microscopy. Cracks in both calcified cartilage and bone may be removed by infilling with calcified matrix. This may be a general repair mechanism for calcified connective tissue crack repair. The fraction of the organ volume occupied by any form of bone tissue in equine distal third metacarpal extremities was increased in the more exercised groups by bone deposited within former marrow adipocytic space. Where deposited upon prior lamellar bone surfaces, this occurred without the intervention of prior resorption and without the formation of a hypermineralized cement line. Exercise inhibited osteoclastic resorption at external anatomical growth modelling sites where it normally occurs. Addition is not coupled to time-wasting resorption: both internally and externally, it occurs both by layering on existing cancellous surfaces and by creation of new immature scaffold, with de novo incorporation of a rich, capillary blood vessel supply. The real response within bone organs subjected to mechanical overload exercise within normal physiological limits is to make more, and to lose less, bone.


The need for specific penalties for hacking in criminal law.
In spite of the fact that hacking is a widely used term, it is still not legally established. Moreover, the definition of the concept of hacking has been deployed in a wide variety of ways in national literature. This ambiguity has led to various side effects. Recently in the United States, reforms collectively known as Aaron's Law were proposed as intended amendments to the Computer Fraud and Abuse Act (CFAA). Most experts expect that this change will put the brakes on the CFAA as a severe punishment policy, and result in a drop in controversial court decisions. In this study, we analyze the definitions and the penalties for hacking for each country and compare them with the national law and then make suggestions through more specific legislation. We expect it will reduce legal controversy and prevent excessive punishment.


Identification, purification, and characterization of an eukaryotic-like phosphopantetheine adenylyltransferase (coenzyme A biosynthetic pathway) in the hyperthermophilic archaeon Pyrococcus abyssi.
Although coenzymeA (CoA) is essential in numerous metabolic pathways in all living cells, molecular characterization of the CoA biosynthetic pathway in Archaea remains undocumented. Archaeal genomes contain detectable homologues for only three of the five steps of the CoA biosynthetic pathway characterized in Eukarya and Bacteria. In case of phosphopantetheine adenylyltransferase (PPAT) (EC 18.104.22.168), the putative archaeal enzyme exhibits significant sequence similarity only with its eukaryotic homologs, an unusual situation for a protein involved in a central metabolic pathway. We have overexpressed in Escherichia coli, purified, and characterized this putative PPAT from the hyperthermophilic archaeon Pyrococcus abyssi (PAB0944). Matrix-assisted laser desorption ionization-time of flight mass spectrometry and high performance liquid chromatography measurements are consistent with the presence of a dephospho-CoA (dPCoA) molecule tightly bound to the polypeptide. The protein indeed catalyzes the synthesis of dPCoA from 4'-phosphopantetheine and ATP, as well as the reverse reaction. The presence of dPCoA stabilizes PAB0944, as it induces a shift from 76 to 82 degrees C of the apparent Tm measured by differential scanning microcalorimetry. Potassium glutamate was found to stabilize the protein at 400 mm. The enzyme behaves as a monomeric protein. Although only distantly related, secondary structure prediction indicates that archaeal and eukaryal PPAT belong to the same nucleotidyltransferase superfamily of bacterial PPAT. The existence of operational proteins highly conserved between Archaea and Eukarya involved in a central metabolic pathway challenge evolutionary scenarios in which eukaryal operational proteins are strictly of bacterial origin.


Online singing groups for people with dementia: scoping review.
In the face of the SARS-CoV-2 pandemic, people with dementia and their carers are contending with serious challenges to their health and wellbeing, due to risk of severe illness, limiting of social contact and disruption to usual activities. Many forms of support for people with dementia and their carers, including singing groups, have moved online using videoconferencing. Previous research has demonstrated the benefits of group singing, which include cognitive stimulation, meaningful activity and peer support. However, although we know which aspects of the singing group experience participants find helpful, we do not know how this experience translates into an online videoconferencing format, and this is a very new field with little existing research. This article reviews the literature pertinent to online singing interventions and uses the findings to develop some suggestions for running an online singing group. SCOPING REVIEW. Systematic literature searches were conducted in EMBASE, Medline, CINAHL, PsycINFO and Web of Science. Owing to the paucity of existing research, searches were also conducted in Google Scholar. The scope of the review covered five related areas: online music making and music therapy, telemedicine and telecare, everyday technology for people with dementia, digital arts and dementia, and use of technology for social interaction and leisure. Our analysis aimed to integrate the results to inform the implementation of online singing groups for people with dementia. Scoping of evidence from discrete fields of enquiry and different disciplinary traditions can inform the delivery of online singing in dementia. This literature also yields useful insights into the role of the carer and how best to support participants to use technology. Barriers and facilitators to online singing were found to relate both to the technology and to the individual participant. Lockdown restrictions have led to much innovation, and this is likely to lead to changes in practice even after normal life resumes. The suggestions in this article will be helpful primarily for practitioners moving into online work and researchers investigating this novel area. They may also be useful to commissioners and policymakers because they reflect current knowledge about best practice.


Plasticity of mesenchymal stem cells in immunomodulation: pathological and therapeutic implications.
Mesenchymal stem cells (MSCs) are multipotent stromal cells that exist in many tissues and are capable of differentiating into several different cell types. Exogenously administered MSCs migrate to damaged tissue sites, where they participate in tissue repair. Their communication with the inflammatory microenvironment is an essential part of this process. In recent years, much has been learned about the cellular and molecular mechanisms of the interaction between MSCs and various participants in inflammation. Depending on their type and intensity, inflammatory stimuli confer on MSCs the ability to suppress the immune response in some cases or to enhance it in others. Here we review the current findings on the immunoregulatory plasticity of MSCs in disease pathogenesis and therapy.


The Relationship between the Physical Activity Environment, Nature Relatedness, Anxiety, and the Psychological Well-being Benefits of Regular Exercisers.
Research from a variety of scientific fields suggests that physical activity in nature and feelings of connection to nature enhance psychological health and well-being. This study investigated the psychological health and well-being impact of the physical activity environment for those already undertaking the recommended weekly amount of physical activity. This topic is important for the design of health and well-being environments and interventions involving physical activity. Participants (N = 262) aged 18-71 years (M = 34.5, SD = 13.1) who met the UK physical activity guidelines completed the Nature Relatedness Scale, the trait section of the State Trait Inventory for Cognitive and Somatic Anxiety and the Psychological Well-Being Scale. Analysis via Multivariate ANOVA indicated that participants who engaged in outdoor physical activity reported significantly lower somatic anxiety levels and higher Nature Relatedness experience (NRexp). Significant results were not evident for wellbeing. Hierarchical regressions revealed that the psychological well-being facet of autonomy, NRexp, and outdoor physical activity predicted lower somatic anxiety, whereas indoor physical activity predicted higher somatic anxiety. Results indicate that somatic anxiety is lower for outdoor physical activity participation, and that outdoor activity, in conjunction with autonomy and NRexp, predicts lower anxiety levels. The findings extend previous work by demonstrating the impact of the physical activity environment on anxiety levels, as well as the contribution of outdoor physical activity and well-being facets to the previously established Nature Relatedness-anxiety relationship.


Protein expression profiling of Coccidioides posadasii by two-dimensional differential in-gel electrophoresis and evaluation of a newly recognized peroxisomal matrix protein as a recombinant vaccine candidate.
Coccidioides posadasii and Coccidioides immitis are dimorphic, soil-dwelling pathogenic ascomycetes endemic to the southwestern United States. Infection can result from inhalation of a very few arthroconidia, but following natural infection, long-lived immunity is the norm. Previous work in the field has shown that spherule-derived vaccines afford more protection than those from mycelia. We have used two-dimensional differential in-gel electrophoresis coupled with nano-high-performance liquid chromatography-tandem mass spectrometry to directly assess both absolute abundance and differential expression of proteins in the spherule and the mycelial phases of C. posadasii with the intent to identify potential vaccine candidates. Peptides derived from 40 protein spots were analyzed and a probable identity was assigned to each. One spherule-abundant protein, identified as Pmp1, showed homology to allergens from Aspergillus fumigatus and other fungi, all of which exhibit similarity to yeast thiol peroxidases. Recombinant Pmp1 was reactive with serum from individuals with both acute and protracted disease, and evoked protection in two murine models of infection with C. posadasii. These results demonstrate the utility of proteomic analysis as a point of discovery for protective antigens for possible inclusion in a vaccine candidate to prevent coccidioidomycosis.


Induced sputum in adolescents with severe stable asthma. Safety and the relationship of cell counts and eosinophil cationic protein to clinical severity.
This study examined the safety of sputum induction and the relation between sputum cell counts and clinical parameters in adolescents with severe persistent asthma. Within 5 days, induced sputum and reversibility in forced expiratory volume in one second (FEV1), quality of life, provocative concentration causing a 20% fall in FEV1 (PC20) of adenosine monophosphate and histamine, exercise-induced bronchoconstriction, overall asthma severity index, and blood eosinophils were collected in 20 atopic adolescents with moderate-to-severe persistent asthma (12-18 yrs of age, FEV1 65-110% of predicted, on 500-2,000 microg inhaled steroids daily). FEV1 was reversible by 13.3-2.3% pred. After sputum induction, FEV1 was still increased by 9.0+/-2.6% pred as compared to the pre-salbutamol baseline. Sputum contained, median (range): 12.4 (0.4-59.5)% squamous cells, 47.3 (6.8-84.0)% macrophages, 39.0 (4.6-84.8)% neutrophils, 4.8 (1.0-12.4)% lymphocytes, 0.4 (0-10.8)% eosinophils and 3.6 (0-23.4)% bronchial epithelial cells. Sputum eosinophils showed a trend towards a significant association with the overall asthma severity index (r=0.46, p=0.06) and correlated inversely with baseline FEV1 (r=-0.51, p=0.03). In conclusion, sputum can be induced safely in adolescents with moderate-to-severe persistent asthma, if pretreated with beta2-agonists. Despite relatively low sputum eosinophil counts in these patients on inhaled steroids, the association of eosinophil numbers with baseline forced expiratory volume in one second and asthma severity index favours a role of induced sputum in monitoring adolescents with severe asthma.


On the onset of motion of sliding drops.
In this article we numerically investigate the onset of motion of liquid drops in contact with a plane and homogeneous substrate with contact angle hysteresis. The drops are driven by a body force F = ρgV, where ρ is the density of the liquid, g is the acceleration of gravity, and V is the volume of the drop. We compare two protocols to vary the bond number Bo = λ(v)/λ(c) by changes of either the drop size λ(v) = V(1/3) or the capillary length λ(c) = (γ/ρg)(1/2) where γ is the interfacial tension, revealing that the transition between pinned and steady moving states can be either continuous or discontinuous. In a certain range both pinned and moving states can be found for a given bond number Bo, depending on the history of the control parameters g and V. Our calculations are extended to arbitrary combinations of static advancing and receding contact angles and provide a comprehensive picture of the depinning transition induced by a quasi-static variation of the control parameters. Finally, we demonstrate that the particular form of the contact line mobility in our model has an impact on the interfacial shape of steady moving drops.


Tele-Rehabilitation for Postural Control by Means of Virtual Reality Rehabilitation System in an Adolescent With Motor Disorder: A Case Study.
The coming of an unforeseen and hostile event such as the COVID-19 pandemic has brought about various changes in everyone's daily life. During the lockdown period, a huge number of restrictions were imposed, hence interrupting a wide range of activities previously proven to be necessary for some people. Due to the circumstances, rehabilitation treatments for children with neurodevelopmental disorders have been suspended, resulting in consequent distress for the children themselves and their parents. To overcome this problem, a tele-rehabilitation approach has proven to be an excellent solution to give continuity to children's rehabilitation. The tele-rehabilitation approach allows access to rehabilitation services directly from home. During lockdown due to the COVID-19 pandemic, the Virtual Reality Rehabilitation System (VRRS) HomeKit, developed by Khymeia, was employed as a rehabilitation system for the treatment of posture and balance of an adolescent with disabilities for the first time ever. Specifically, a 17-year-old female patient was enrolled and evaluated by the clinical staff before and after the home-training sessions. The system was delivered to the patient's home so that she could perform the tasks in a familiar environment, while under the supervision of the clinical staff. Through a specific platform, using different modules of the system, therapists could remotely check that the proposed exercises were properly performed and provide feedback and/or increase the difficulty according to the patient's needs and progress. Therefore, the treatment performed was carried out at home in a personalized, intensive, and playful way; characteristics do not present in a traditional treatment. Our results are promising and demonstrate both the efficacy of rehabilitation exercises carried out at home and the feasibility of home-based rehabilitation, when using the VRRS HomeKit even with adolescents. The VRRS HomeKit presents some limitations, such as the need to have the line connection and free space at home, the presence of technical issues, and the education of parents and patients to understand the instructions. Despite the limitations, this study provides the basis for continuing the experience of tele-rehabilitation on patients with a motor disorder also by customizing the exercises to their characteristics.


Feline immunodeficiency virus cell entry.
The process of feline immunodeficiency virus (FIV) cell entry was examined using assays for virus replication intermediates. FIV subtype B was found to utilize the chemokine receptor CXCR4, but not CCR5, as a cellular receptor. Zidovudine blocked formation of late viral replication products most effectively, including circular DNA genome intermediates. Our findings extend the role of CXCR4 as a primary receptor for CD4-independent cell entry by FIV.


Update on Surgical Management of Pediatric Urolithiasis.
Urolithiasis has always been a fascinating disease, even more so in children. There are many intriguing facets to this pathology. This article is a nonsystematic review to provide an update on the surgical management of pediatric urolithiasis. It highlights the pros and cons of various minimally invasive surgical options such as extracorporeal shockwave lithotripsy (ESWL), retrograde intrarenal surgery (RIRS), percutaneous nephrolithotomy (PCNL), laparoscopy, and robotics. This article also describes the various intracorporeal disintegration technologies available to fragment the stone, including the newer advancements in laser technology. It also emphasizes the cost considerations especially with reference to the emerging economies. Thus, this manuscript guides how to select the least-invasive option for an individual patient, considering age and gender; stone size, location, and composition; and facilities and expertise available.


Age Differences among Female Sex Workers in the Philippines: Sexual Risk Negotiations and Perceived Manager Advice.
Consistent condom use among high risk groups such as female sex workers (FSWs) remains low. Adolescent female sex workers are especially at higher risk for HIV/STI infections. However, few published studies have compared the sexual risk negotiations among adolescent, emerging adult, and older age groups or the extent a manager's advice about condom use is associated with an FSW's age. Of 1,388 female bar/spa workers surveyed in the southern Philippines, 791 FSW who traded sex in the past 6 months were included in multivariable logistic regression models. The oldest FSWs (aged 36-48) compared to adolescent FSWs (aged 14-17) were 3.3 times more likely to negotiate condoms when clients refused condom use. However, adolescent FSWs received more advice from their managers to convince clients to use condoms or else to refuse sex, compared to older FSWs. Both adolescent and the oldest FSWs had elevated sexually transmitted infections (STIs) and inconsistent condom use compared to other groups. Having a condom rule at the establishment was positively associated with condom negotiation. Factors such as age, the advice managers give to their workers, and the influence of a condom use rule at the establishment need to be considered when delivering HIV/STI prevention interventions.


Development of Onchocerca volvulus larvae in Simulium pintoi in the Amazonas region of Venezuela.
The intake and development of Onchocerca volvulus in Simulium pintoi from the Parima mountain region of the Federal Territory of Amazonas in Venezuela, were studied experimentally. When wild females fed on the lower half of the legs and lower third of the back of an onchocerciasis patient harboring 23 and 264 microfilariae per skin snip, respectively, at each site, an average (median) of 14 (range, 1-77) and 245 (range, 58-495) microfilariae was ingested. However, within 24 hours of microfilarial ingestion a mortality of 47% (16/34 flies) was observed in the group of flies which fed on the back, as compared with 2% (2/101 flies) in the other group which fed on the legs. At a temperature varying between 16 degrees C and 24 degrees C, the development of O. volvulus larvae in S. pintoi was synchronous and orderly; no abnormal nor deformed larvae were observed. Third-stage larvae were first seen in the head of flies dying between 8 and 9 days after microfilarial ingestion, and 98 of 100 larvae recovered from days 10-16 were in the third stage. The proportions of females harboring third-stage larvae among flies which lived through day 8 in the two groups which fed on the legs and back, respectively, were 55% (21/38 flies) and 63% (5/8 flies). Although only two of five positive flies in the latter group contained third-stage larvae in the head (1 and 12, respectively), 71% (15/21 positive flies) of the former group had an average of 2.7 third-stage larvae in the head (range, 1-10).(ABSTRACT TRUNCATED AT 250 WORDS)


Transient increase in HDL-cholesterol during weight gain by hyperalimentation in healthy subjects.
Determination of lipid levels is fundamental in cardiovascular risk assessment. We studied the short-term effects of fast food-based hyperalimentation on lipid levels in healthy subjects. Twelve healthy men and six healthy women with a mean age of 26 ± 6.6 years and an aged-matched control group were recruited for this prospective interventional study. Subjects in the intervention group aimed for a body weight increase of 5-15% by doubling the baseline caloric intake by eating at least two fast food-based meals a day in combination with adoption of a sedentary lifestyle for 4 weeks. This protocol induced a weight gain from 67.6 ± 9.1 kg to 74.0 ± 11 kg (P < 0.001). A numerical increase in the levels of high-density lipoprotein (HDL)-cholesterol occurred in all subjects during the study and this was apparent already at the first week in 16/18 subjects (mean increase at week 1: +22.0 ± 16%, range from -7 to +50%), whereas the highest level of HDL during the study as compared with baseline values varied from +6% to +58% (mean +31.6 ± 15%). The intake of saturated fat in the early phase of the trial related positively with the HDL-cholesterol-increase in the second week (r = 0.53, P = 0.028). Although the levels of insulin doubled at week 2, the increase in low-density lipoprotein (LDL)-cholesterol was only +12 ± 17%, and there was no statistically significant changes in fasting serum triglycerides. We conclude that hyperalimentation can induce a fast but transient increase in HDL-cholesterol that is of clinical interest when estimating cardiovascular risk based on serum lipid levels.


Prevalence and Spectrum of Gastro Esophageal Reflux Disease in Bronchial Asthma.
There exists a complex interplay between asthma and gastroesophageal reflux disease. Both these diseases are known to aggravate each other and amelioration of one is necessary for the control of the other. There is a paucity of studies in Indian population on this subject. To evaluate the clinical features and the endoscopic findings of the upper gastrointestinal tract in patients with bronchial asthma. Study was conducted at KMC group of hospitals, Mangalore in the Department of chest medicine in association with Department of gastroenterology. Subjects included 50 cases of bronchial asthma and controls were 58 non asthmatic patients with allergic rhinitis and chronic urticaria. All patients were queried about presence or absence of symptoms of upper gastro intestinal tract disorders by gastro oesophageal reflux disease (GERD) questionnaire and all the included patients underwent upper gastro intestinal endoscopy. The study showed that symptoms of gastroesophageal reflux were significantly more in asthmatics (52%) as compared to the controls (28%). The common presenting features of gastroesophageal reflux in asthmatics were heartburn (40%) retrosternal pain (24%), nocturnal cough (18%), dyspepsia (16%) and regurgitation (14%) and the above symptoms were significantly more common in asthmatics as compared to controls. Gastroesophageal reflux disease was found to be significantly more common in the asthmatics (58%) as compared to the control group where it was present in 32.75% of the subjects. Clinical or endoscopic evidence of any upper gastrointestinal disorder was found in 68% of the asthmatics as compared to 37.93% of the controls. This difference was found to be statistically significant. The study showed that gastroesophageal reflux disease was significantly more in asthmatics as compared to the controls. Upper gastrointestinal symptoms were more common in asthmatics as against controls. Clinical or endoscopic evidence of upper gastrointestinal disorder and gastroesophageal reflux disease was found in significantly higher proportion of the asthmatics as compared to the controls. Clinically silent gastroesophageal reflux disease was however seen in both control and asthmatic groups equally with a lower prevalence.


Burkholderia pseudomallei Capsule Exacerbates Respiratory Melioidosis but Does Not Afford Protection against Antimicrobial Signaling or Bacterial Killing in Human Olfactory Ensheathing Cells.
Melioidosis, caused by the bacterium Burkholderia pseudomallei, is an often severe infection that regularly involves respiratory disease following inhalation exposure. Intranasal (i.n.) inoculation of mice represents an experimental approach used to study the contributions of bacterial capsular polysaccharide I (CPS I) to virulence during acute disease. We used aerosol delivery of B. pseudomallei to establish respiratory infection in mice and studied CPS I in the context of innate immune responses. CPS I improved B. pseudomallei survival in vivo and triggered multiple cytokine responses, neutrophil infiltration, and acute inflammatory histopathology in the spleen, liver, nasal-associated lymphoid tissue, and olfactory mucosa (OM). To further explore the role of the OM response to B. pseudomallei infection, we infected human olfactory ensheathing cells (OECs) in vitro and measured bacterial invasion and the cytokine responses induced following infection. Human OECs killed >90% of the B. pseudomallei in a CPS I-independent manner and exhibited an antibacterial cytokine response comprising granulocyte colony-stimulating factor, tumor necrosis factor alpha, and several regulatory cytokines. In-depth genome-wide transcriptomic profiling of the OEC response by RNA-Seq revealed a network of signaling pathways activated in OECs following infection involving a novel group of 378 genes that encode biological pathways controlling cellular movement, inflammation, immunological disease, and molecular transport. This represents the first antimicrobial program to be described in human OECs and establishes the extensive transcriptional defense network accessible in these cells. Collectively, these findings show a role for CPS I in B. pseudomallei survival in vivo following inhalation infection and the antibacterial signaling network that exists in human OM and OECs.


Pattern of ocular dominance columns and cytochrome oxidase activity in a macaque monkey with naturally occurring anisometropic amblyopia.
Unilateral eyelid suture, a model for amblyopia induced by congenital cataract, produces shrinkage of the deprived eye's ocular dominance columns in the striate cortex. Loss of geniculocortical projections are thought to account for the poor vision in the amblyopic eye. It is uncertain whether ocular dominance columns become shrunken in other forms of amblyopia. We examined the striate cortex in a pigtailed macaque with natural anisometropia discovered at age 5 months. Amblyopia in the left eye was documented at 1 year by behavioral testing. At age 6 years, the left eye was injected with [3H]proline and the striate cortex was processed for autoradiography and cytochrome oxidase (CO). The ocular dominance columns in layer IVc labelled with [3H]proline were normal. CO staining showed a novel pattern of thin dark bands in layer IV. These bands occupied the core zones at the center of the ocular dominance columns. Their appearance resulted from relative loss of CO activity along the borders of the ocular dominance columns, regions specialized for binocular processing. These findings indicate that not all forms of amblyopia are accompanied by shrinkage of ocular dominance columns. The unusual pattern of CO staining in layer IVc reflected a subtle alteration in metabolic activity which may have resulted from impairment of binocular function in anisometropic amblyopia.


Emergent collective dynamics of bottom-heavy squirmers under gravity.
We present the results of hydrodynamic simulations using the method of multi-particle collision dynamics for a system of squirmer microswimmers moving under the influence of gravity at low Reynolds numbers. In addition, the squirmers are bottom-heavy so that they experience a torque which aligns them along the vertical. The squirmers interact hydrodynamically by the flow fields of a stokeslet and rotlet, which are initiated by the acting gravitational force and torque, respectively, and by their own flow fields. By varying the ratio of swimming to bulk sedimentation velocity and the torque, we determine state diagrams for the emergent collective dynamics of neutral squirmers as well as strong pushers and pullers. For low swimming velocity and torque we observe conventional sedimentation, while the sedimentation profile becomes inverted when their values are increased. For neutral squirmers we discover convective rolls of circulating squirmers between both sedimentation states, which sit at the bottom of the system and are fed by plumes made of collectively sinking squirmers. At larger torques porous clusters occur that spawn single squirmers. The two latter states can also occur transiently starting from a uniform squirmer distribution and then disappear in the long-time limit. For strong pushers and pullers only weak plume formation is observed.


A new, easy-to-make pectin-honey hydrogel enhances wound healing in rats.
Honey, alone or in combination, has been used for wound healing since ancient times and has reemerged as a topic of interest in the last decade. Pectin has recently been investigated for its use in various biomedical applications such as drug delivery, skin protection, and scaffolding for cells. The aim of the present study was to develop and evaluate a pectin-honey hydrogel (PHH) as a wound healing membrane and to compare this dressing to liquid honey. Thirty-six adult male Sprague-Dawley rats were anesthetized and a 2 × 2 cm excisional wound was created on the dorsum. Animals were randomly assigned to four groups (PHH, LH, Pec, and C): in the PHH group, the pectin-honey hydrogel was applied under a bandage on the wound; in the LH group, liquid Manuka honey was applied; in the Pec group, pectin hydrogel was applied (Pec); and in the C group, only bandage was applied to the wound. Images of the wound were taken at defined time points, and the wound area reduction rate was calculated and compared between groups. The wound area reduction rate was faster in the PHH, LH, and Pec groups compared to the control group and was significantly faster in the PHH group. Surprisingly, the Pec group exhibited faster wound healing than the LH group, but this effect was not statistically significant. This is the first study using pectin in combination with honey to produce biomedical hydrogels for wound treatment. The results indicate that the use of PHH is effective for promoting and accelerating wound healing.


Neuroregenerative Effect of Oxandrolone: A Case Report.
Anabolic steroids have the clinical effect of increasing protein synthesis in muscle and other tissues. The brain and spinal cord neurons have gonadal steroid receptors and various studies have shown at structural and molecular levels that androgenic steroids have a significant trophic effect on the brain and spinal cord. We evaluated the effect of Oxandrolone (an FDA-approved anabolic steroid) at the dose of 20 mg/day for 3 months added to concomitant exercise strength training 3 times a week in a patient affected by a demyelinating disease, Charcot-Marie-Toot 1 (CMT1). After the treatment, an increase in muscular strength and walking capacity was observed. Muscle biopsy revealed a significant increase of type grouping of muscle fibers, an expression of regeneration and reinnervation processes. Data ensuing from this single case-report suggest that anabolic androgenic steroids have a potential neuroregenerative effect, with an inherent improvement in neuromuscular efficiency through an increased myelin synthesis at peripheral nervous system site.


Effective induction of cell death on adult T-cell leukaemia cells by HLA-DRbeta-specific small antibody fragment isolated from human antibody phage library.
By a biopanning method using cell sorter, we quickly isolated an antibody phage clone (S1T-A3) specific to human T-lymphotropic virus type 1-carrying T-cell line S1T from a human single chain Fv (scFv) antibody phage library. This scFv antibody bound to HTLV-1-carrying T-cell lines including MT-2, MT-4 and M8166 other than S1T, but not to non-HTLV-1-carrying T-cell lymphomas such as Jurkat and MOLT4 cells. Interestingly, this antibody induced the cell death on S1T cells very quickly (< 30 min). We tried to identify the target molecules by western blotting and mass spectrometric analysis, revealing that the target antigen was HLA class II DR. The cell death was induced only in dimmer form of scFv (diabody) and at 15-fold lower concentration than that of a fusion protein of scFv and human IgG Fc [(scFv)(2)-Fc] or anti HLA-DR mouse whole antibody L243. Thus, S1T-A3 diabody is a small antibody fragment with agonistic activity to induce cell death through HLA-DR. This is the first report elucidating that diabody specific to HLA-DR is effective to induce the cell death in T-cell malignancy especially adult T-cell leukaemic cell line.


Gene disruption of ribosomal protein L5 (RPL5) decreased the sensitivity of CHO-K1 cells to uncoupler carbonylcyanide-3-chlorophenylhydrazone.
Protonophoric uncoupler carbonylcyanide-3-chlorophenylhydrazone (CCCP) decreases the proton motive force (ΔP) of the mitochondrial inner membrane and results in inhibition of oxidative phosphorylation. In this study, a CCCP-resistant clone was isolated from a random gene trap insertional mutant library of Chinese hamster ovary (CHO)-K1 cells which was constructed by infecting a retrovirus vector, ROSAβgeo. Although we expected the isolation of the mutants defective in nuclear genes responsible for mitochondrial functions, the disrupted gene of the isolated mutant that we named R1 cells was identified as one of the alleles for ribosomal protein 5 of large subunit (RPL5). The R1 cells express as much as 80% RPL5 protein compared with the parental CHO-K1 cells, possibly due to enhanced transcription from a remaining wild-type RPL5 allele in R1 cells. Furthermore, the protein amount is not decreased by CCCP in R1 cells, in contrast to its clear reduction by CCCP in parental cells. Since mutations of RPL5 and other ribosomal proteins are responsible for the ribosomopathies and cancer, the present mutant may be a useful cellular model of such human diseases from a viewpoint of energy metabolism as well as a tool for the study of ribosome biogenesis and extra-ribosomal function of the RPL5 protein.


Epilepsy in Asia: Disease burden, management barriers, and challenges.
This article reviews the burden of epilepsy in Asia, the challenges faced by people with epilepsy, and the management of epilepsy. Comparison is made with other parts of the world. For this narrative review, data were collected using specified search criteria. Articles investigating the epidemiology of epilepsy, diagnosis, comorbidities and associated mortality, stigmatization, and treatment were included. Epilepsy is a global health care issue affecting up to 70 million people worldwide. Nearly 80% of people with epilepsy live in low- and middle-income countries with limited resources. People with epilepsy are prone to physical and psychological comorbidities, including anxiety and depression, which can negatively impact their quality of life. Furthermore, people with epilepsy are at higher risk of premature death than people without epilepsy. Discrimination or stigmatization of people with epilepsy is common in Asia and can affect their education, work, and marriage opportunities. Access to epilepsy treatment varies throughout Asia. Although highly advanced treatment is available in some countries, up to 90% of people with epilepsy are not adequately treated or are not treated with conventional antiepileptic therapy in resource-limited countries. People in remote areas often do not receive any epilepsy care. First-generation antiepileptic drugs (AEDs) are available, but usually only in urban areas, and second-generation AEDs are not available in all countries. Newer AEDs tend to have more favorable safety profiles than first-generation AEDs and provide options to tailor therapy for individual patients, especially those with comorbidities. Active epilepsy surgery centers are present in some countries, although epilepsy surgery is often underutilized given the number of patients who could benefit. Further epidemiologic research is needed to provide accurate epilepsy data across the Asian region. Coordinated action is warranted to improve access to treatment and care.


Humic acid induces apoptosis in human premyelocytic leukemia HL-60 cells.
It has been shown that humic acid (HA), a phenolic polymer, exhibits pro-oxidant and cytotoxic effects. In this study, HA induction of apoptosis was studied using cultured human premyelocytic leukemia HL-60 cells. Treatment at a range of HA concentrations (50-400 microg/ml) resulted in dose-and time-dependent sequences of events marked by apoptosis, as demonstrated through by apoptotic features such as loss of cell viability, chromatin condensation, and internucleosomal DNA fragmentation. This HA-induced apoptosis in the HL-60 cells was mainly associated with the release of cytochrome c from the mitochondria. Furthermore, apoptosis in the HL-60 cells was accompanied by the activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase (PARP), a major component in the apoptotic cell death mechanism. Although the HA-induced apoptosis was associated with Bax protein levels, negligible Bcl-2 reduction was observed. Analysis of the data reported herein reveals that HA exerts antiproliferative action and growth inhibition on HL-60 cells through induction of apoptosis, which may have anticancer properties potentially useful for the development of new drug products.


The M235T variant of the angiotensinogen gene and the body mass index are useful markers for prevention of hypertension in pregnancy: a tree-based analysis of gene-environment interaction.
We sought to perform a tree-based analysis of lifestyle risk factors for hypertension in pregnancy (HP) with univariate and multivariate analyses. Seventy-eight HP patients and 199 normal controls were recruited from primiparous women 20 to 34 years of age. Data from angiotensinogen (AGT) genotyping and data from a self-administered questionnaire about lifestyle were subjected to univariate and multivariate analyses. By dividing the subjects into two subgroups--those who possessed "the TT genotype of AGT" and "body mass index (BMI) < 24" and those who did not--we were able to examine the acquired risk factors for HP during pregnancy in these two groups. Multivariate analysis selected "mentally stressful condition" and "no antenatal training during pregnancy" in the former group, and "poorly balanced diet" in the latter group. Determination of factors obvious before pregnancy, such as genotype or prepregnancy BMI, may be useful for devising effective individualized strategies for preventing HP.


Where have all the interactions gone? Estimating the coverage of two-hybrid protein interaction maps.
Yeast two-hybrid screens are an important method for mapping pairwise physical interactions between proteins. The fraction of interactions detected in independent screens can be very small, and an outstanding challenge is to determine the reason for the low overlap. Low overlap can arise from either a high false-discovery rate (interaction sets have low overlap because each set is contaminated by a large number of stochastic false-positive interactions) or a high false-negative rate (interaction sets have low overlap because each misses many true interactions). We extend capture-recapture theory to provide the first unified model for false-positive and false-negative rates for two-hybrid screens. Analysis of yeast, worm, and fly data indicates that 25% to 45% of the reported interactions are likely false positives. Membrane proteins have higher false-discovery rates on average, and signal transduction proteins have lower rates. The overall false-negative rate ranges from 75% for worm to 90% for fly, which arises from a roughly 50% false-negative rate due to statistical undersampling and a 55% to 85% false-negative rate due to proteins that appear to be systematically lost from the assays. Finally, statistical model selection conclusively rejects the Erdös-Rényi network model in favor of the power law model for yeast and the truncated power law for worm and fly degree distributions. Much as genome sequencing coverage estimates were essential for planning the human genome sequencing project, the coverage estimates developed here will be valuable for guiding future proteomic screens. All software and datasets are available in and , -, and -, and are also available from our Web site, http://www.baderzone.org.