[Plasma ascorbic acid: preanalytical factors and new HPLC method].
We describe a fast ED-HPLC method for ascorbic acid analysis, carried out on a new mixed mode chromatographic column. Due to its functional dodecylsilane group associating a quaternary ammonium, this column is simpler and easier to handle than previous ones because without any counter-ion in the mobile phase. As ascorbic acid and glutathione are very often measured on the same blood sample, it was efficient to validate a common pretreatment solution for ascorbic acid in plasma and glutathione in whole blood. This pretreatment is based on the use of a sulfosalycilic acid, N-ethylmaleimide, EDTA mix recently described. Plasma samples, stored after deproteinisation and without centrifugation, are stable for three weeks at -20 or -80 degrees C.


Peripheral nerve injury and Raynaud's syndrome following electric shock.
A truck driver was injured by a high-voltage line of 10,000 volts when holding a metallic bar in both hands. Initially no neurological abnormalities were found, but during the following few weeks increasing sensory and minor motor symptoms developed in the right upper extremity. After one year numbness of the right thigh and leg appeared, as well as attacks of white finger in both hands. Repeated examinations showed progressive abnormalities of the median and ulnar nerves in both hands. No other cause for Raynaud's syndrome was discovered. The late high-voltage effects, presumably indirect, are suggested to be of multifactorial etiology.


Effect of superstimulation protocols on nuclear maturation and distribution of lipid droplets in bovine oocytes.
Our objective was to study the effect of superstimulation protocols on nuclear maturation of the oocyte and the distribution of lipid droplets in the ooplasm. Heifers (n=4 each group) during the luteal phase were either treated with FSH for 4 days (Short FSH), FSH for 4 days followed by 84h of gonadotropin free period (FSH Starvation) or for 7 days (Long FSH) starting from the day of wave emergence. In all groups, LH was given 24h after induced luteolysis (penultimate day of FSH) and cumulus-oocyte complexes were collected 24h later. Oocytes were stained for nuclear maturation (Lamin/chromatin) and lipid droplets (Nile red). The Long FSH group had a greater proportion of mature oocytes (metaphase II) compared with heifers in the Short FSH and FSH Starvation groups (59/100 vs 5/23 and 2/25, respectively; P<0.01). On average across all groups, oocytes contained 22pL of lipids (3.3% of ooplasm volume) distributed as 3000 droplets. Average volume of individual lipid droplets was higher in the FSH Starvation (11.5±1.5 10(-3) pL, P=0.03) compared with the Short and Long FSH groups (7.2±0.6 10(-3) and 8.0±0.8 10(-3) pL, respectively). In conclusion, both FSH Starvation and Short FSH treatments yielded a lower proportion of mature oocytes compared with the Long FSH treatment. Furthermore, FSH starvation led to an accumulation of larger lipid droplets in the ooplasm, indicating atresia. Our results indicate that a longer superstimulation period in beef cattle yields higher numbers and better-quality oocytes.


Incorporation of memory effects in coarse-grained modeling via the Mori-Zwanzig formalism.
The Mori-Zwanzig formalism for coarse-graining a complex dynamical system typically introduces memory effects. The Markovian assumption of delta-correlated fluctuating forces is often employed to simplify the formulation of coarse-grained (CG) models and numerical implementations. However, when the time scales of a system are not clearly separated, the memory effects become strong and the Markovian assumption becomes inaccurate. To this end, we incorporate memory effects into CG modeling by preserving non-Markovian interactions between CG variables, and the memory kernel is evaluated directly from microscopic dynamics. For a specific example, molecular dynamics (MD) simulations of star polymer melts are performed while the corresponding CG system is defined by grouping many bonded atoms into single clusters. Then, the effective interactions between CG clusters as well as the memory kernel are obtained from the MD simulations. The constructed CG force field with a memory kernel leads to a non-Markovian dissipative particle dynamics (NM-DPD). Quantitative comparisons between the CG models with Markovian and non-Markovian approximations indicate that including the memory effects using NM-DPD yields similar results as the Markovian-based DPD if the system has clear time scale separation. However, for systems with small separation of time scales, NM-DPD can reproduce correct short-time properties that are related to how the system responds to high-frequency disturbances, which cannot be captured by the Markovian-based DPD model.


[Histomorphology of secondary cartilage in human fetal mandibles].
The aim of this study was to provide a histomorphological analysis of some secondary cartilages of mandible and temporal bone as observed in human fetuses 18-22 weeks old. The behavior of cartilage was studied in both these regions, which were decalcified, cut at 10 mu, stained with Mallory staining and examined by optical microscopy. In mandible symphysis menti and condylar cartilage were described. The symphysis appeared to be formed by a fibrous cartilagineous structure surrounded by membranous bone. This structure seems be round in the caudal sections and ovoidal in the rostral sections with the major axis perpendicular to the mean sagittal plane. Meanwhile the condyle is formed by secondary cartilage which may be appreciated in this development stage 5 zona. Secondary cartilage was observed also in the temporal bone nearby the primitive glenoid fossa. The development and the importance of these cartilagineous structures are discussed.


Vector graph assisted pedestrian dead reckoning using an unconstrained smartphone.
The paper presents a hybrid indoor positioning solution based on a pedestrian dead reckoning (PDR) approach using built-in sensors on a smartphone. To address the challenges of flexible and complex contexts of carrying a phone while walking, a robust step detection algorithm based on motion-awareness has been proposed. Given the fact that step length is influenced by different motion states, an adaptive step length estimation algorithm based on motion recognition is developed. Heading estimation is carried out by an attitude acquisition algorithm, which contains a two-phase filter to mitigate the distortion of magnetic anomalies. In order to estimate the heading for an unconstrained smartphone, principal component analysis (PCA) of acceleration is applied to determine the offset between the orientation of smartphone and the actual heading of a pedestrian. Moreover, a particle filter with vector graph assisted particle weighting is introduced to correct the deviation in step length and heading estimation. Extensive field tests, including four contexts of carrying a phone, have been conducted in an office building to verify the performance of the proposed algorithm. Test results show that the proposed algorithm can achieve sub-meter mean error in all contexts.


Blood flow determinations utilizing digital densitometry.
A method of obtaining relative and absolute blood flow measurements from digital densitometry was evaluated with a simulated vessel phantom and a hydrodynamic model. A digital vascular imaging system capable of acquisition in 512(2) and 1024(2) mode was used. Relative and absolute blood flow were measured using parameters derived from the densitometric curve. Since application of densitometric data to absolute flow measurements requires the vessel diameter, an algorithm for vessel size determination was created. Gray scale changes were demonstrated to be linearly related to contrast concentration. The variance of vessel size determination was significantly different in all combinations of 1024(2) and 512(2) imaging with 15 cm or 35 cm field size. The error in vessel size determination was significantly less using the larger 1024(2) matrix and the smaller 15 cm image intensifier field size, as shown by the smaller variance. In relative flow determinations, there was good correlation between the flow and four parameters of the densitometric curve with no significant differences between 512(2) and 1024(2) imaging. Absolute flow determinations had slightly lower correlation to actual flow but were not significantly different from relative flow determinations. Relative and absolute blood flow determinations can be performed adequately with either 512(2) or 1024(2) imaging. The increased accuracy in vessel size determination with 1024(2) imaging makes this high resolution system potentially preferable to determine absolute blood flow.


Monitoring and comorbidities in patients with chronic hepatitis B currently treated with nucleos(t)ide analogs.
Long-term monotherapy with nucleos(t)ide analogs (NAs) represents the treatment option for the majority of patients with chronic hepatitis B (CHB), an aging population with a greater likelihood of comorbidities. We assessed the prevalence of concurrent non-hepatic diseases and the safety monitoring in a large cohort of CHB patients receiving NAs and their potential impact on disease outcomes. We included 500 consecutive CHB patients from 5 major tertiary Greek centers, under long-term therapy with an NA. Epidemiological/clinical characteristics and data on concomitant disease, drug use and investigations ordered were collected. The mean age was 58 years and 66% were male. Most patients were receiving tenofovir disoproxil fumarate (TDF, 60%) or entecavir (ETV, 37%) monotherapy. Decompensated cirrhosis at baseline was present in 10%, while hepatocellular carcinoma (HCC) under therapy developed in 21 patients. The median duration of total NA therapy was 56 and of latest therapy 42 months. The most common (prevalence >10%) comorbidities were hypertension (28%), non-HCC cancer(s) (12%), and diabetes (11%). Patients with a longer duration of latest therapy (≥4 vs. <4 years) were older (mean age: 58 vs. 56 years, P=0.004), had more frequent history of prior use of NA(s) (53% vs. 35%, P<0.001), and less frequent liver decompensation (5% vs. 13%, P=0.008) and non-HCC cancers (8% vs. 15%, P=0.020). HCC developed more frequently in patients with than in those without diabetes (11% vs. 3%, P=0.022). Greek CHB patients currently treated with NAs, almost exclusively ETV or TDF, are often older than 60 years, have several comorbidities, and thus require careful management.


Honey Bees and Neonicotinoid-Treated Corn Seed: Contamination, Exposure, and Effects.
Most corn (Zea mays) seeds planted in the United States in recent years are coated with a seed treatment containing neonicotinoid insecticides. Abrasion of the seed coating generates insecticide-laden planter dust that disperses through the landscape during corn planting and has resulted in many "bee-kill" incidents in North America and Europe. We investigated the linkage between corn planting and honey bee colony success in a region dominated by corn agriculture. Over 3 yr we consistently observed an increased presence of corn seed treatment insecticides in bee-collected pollen and elevated worker bee mortality during corn planting. Residues of seed treatment neonicotinoids, clothianidin and thiamethoxam, detected in pollen positively correlated with cornfield area surrounding the apiaries. Elevated worker mortality was also observed in experimental colonies fed field-collected pollen containing known concentrations of corn seed treatment insecticides. We monitored colony growth throughout the subsequent year in 2015 and found that colonies exposed to higher insecticide concentrations exhibited slower population growth during the month of corn planting but demonstrated more rapid growth in the month following, though this difference may be related to forage availability. Exposure to seed treatment neonicotinoids during corn planting has clear short-term detrimental effects on honey bee colonies and may affect the viability of beekeeping operations that are dependent on maximizing colony size in the springtime. Environ Toxicol Chem 2021;40:1212-1221. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.


Periventricular leucomalacia and intraventricular haemorrhage in the preterm neonate.
Two hundred very low birthweight infants were prospectively scanned to ascertain the incidence of periventricular leucomalacia (PVL) and haemorrhage. Before collection of data, clear definitions of ultrasound abnormalities believed to represent PVL and intraventricular haemorrhage were described. These referred to small and moderate intraventricular haemorrhage, paenchymal haemorrhage, and PVL, including prolonged flare (echoes in the periventricular region lasting for two weeks or more and not becoming cystic). Sixty nine infants (34%) had no abnormality on ultrasound scans. Intraventricular haemorrhage occurred in 107 babies (37 grade I and 62 grade II), and only eight infants were thought to have true parenchymal haemorrhage. Ultrasound appearances of PVL were seen in 27 infants, 19 of whom developed cysts and eight died in the precystic stage. Prolonged flare occurred in another 25 babies. Unilateral parenchymal haemorrhage occurred in four infants who subsequently developed cystic PVL in the contralateral hemisphere. Twenty one infants developed ventricular dilatation, 12 of whom had associated parenchymal lesions. Haemorrhage, PVL, and flare occurred commonly in infants of 30 weeks' gestation and below and became markedly less common in more mature infants. We believe prolonged flare represents a form of PVL, and in this study a total of 52 (26%) infants had an ultrasound appearance of periventricular leucomalacia, an incidence considerably higher than previously reported.


Whole genome sequencing reveals extended natural transformation in Campylobacter impacting diagnostics and the pathogens adaptive potential.
Campylobacter is the major bacterial agent of human gastroenteritis worldwide and represents a crucial global public health burden. Species differentiation of C. jejuni and C. coli and phylogenetic analysis is challenged by inter-species horizontal gene transfer. Routine real-time PCR on more than 4000 C. jejuni and C. coli field strains identified isolates with ambiguous PCR results for species differentiation, in particular, from the isolation source eggs. K-mer analysis of whole genome sequencing data indicated the presence of C. coli hybrid strains with huge amounts of C. jejuni introgression. Recombination events were distributed over the whole chromosome. MLST typing was impaired, since C. jejuni sequences were also found in six of the seven housekeeping genes. cgMLST suggested that the strains were phylogenetically unrelated. Intriguingly, the strains shared a stress response set of C. jejuni variant genes, with proposed roles in oxidative, osmotic and general stress defence, chromosome maintenance and repair, membrane transport, cell wall and capsular biosynthesis and chemotaxis. The results have practical impact on routine typing and on the understanding of the functional adaption to harsh environments, enabling successful spreading and persistence of Campylobacter.


Fluorescent labelling of in situ hybridisation probes through the copper-catalysed azide-alkyne cycloaddition reaction.
In situ hybridisation is a powerful tool to investigate the genome and chromosome architecture. Nick translation (NT) is widely used to label DNA probes for fluorescence in situ hybridisation (FISH). However, NT is limited to the use of long double-stranded DNA and does not allow the labelling of single-stranded and short DNA, e.g. oligonucleotides. An alternative technique is the copper(I)-catalysed azide-alkyne cycloaddition (CuAAC), at which azide and alkyne functional groups react in a multistep process catalysed by copper(I) ions to give 1,4-distributed 1,2,3-triazoles at a high yield (also called 'click reaction'). We successfully applied this technique to label short single-stranded DNA probes as well as long PCR-derived double-stranded probes and tested them by FISH on plant chromosomes and nuclei. The hybridisation efficiency of differently labelled probes was compared to those obtained by conventional labelling techniques. We show that copper(I)-catalysed azide-alkyne cycloaddition-labelled probes are reliable tools to detect different types of repetitive sequences on chromosomes opening new promising routes for the detection of single copy gene. Moreover, a combination of FISH using such probes with other techniques, e.g. immunohistochemistry (IHC) and cell proliferation assays using 5-ethynyl-deoxyuridine, is herein shown to be easily feasible.


[Inspiratory muscle resistance to fatigue during exercise and simulated airway obstruction].
Respiratory muscle fatigue can develop during simulated airway obstruction. The aim of this study was to characterize the pattern of inspiratory muscle fatigue and to assess the resistance to fatigue of diaphragm (D), parasternal (PS), sternocleidomastoid (SM) and scalene (SC). 8 healthy untrained subjects participated in this study. To identify signs of inspiratory muscles fatigue development electromyographic activity of D, PS, SCM and SC was recorded during 5-min exercise with loaded breathing (40 cm H2O/L · s(-1)). The before-to-after exercise measurements of maximal inspiratory pressure (MIP) and EMG power spectrum changes were performed. Maximal inspiratory pressure declined about 12% after exercise test compared with control, whereas the peak magnitude of integrated electrical activity of D, PS, SCM and SC during post-exercise Muller's maneuver was significantly greater than in pre-exercise test in all subjects. The extent ofinspiratory muscles fatigue was evaluated by analysis of shift in centroid frequency (fc) of EMG power spectrum. All subjects demonstrated a significant reduction in fc of PS, SCM and SC.fc of D was not changed. Diaphragm is more resistantto fatigue during obstructive breathing compared with PS, SCM and SC. The data suggest that the reduction of maximum inspiratory pressure in chronic obstructive pulmonary disease also caused primarily by the weakening of the accessory muscles, while the weakness of the diaphragm may occur in the later stages of the disease. The functional failure of accessory muscles is an additional factor, which, along with the additional breathing resistance increases the load on the diaphragm, promoting its fatigue and reduced respiratory reserve.


A pilot study using carboplatin, vincristine, and temozolomide in children with progressive/symptomatic low-grade glioma: a Children's Oncology Group study†.
This study was initiated to test the feasibility and toxicity of a regimen that alternates the administration of weekly carboplatin and vincristine with temozolomide in the management of children with progressive and/or symptomatic low-grade glioma. Eligible children received a 10-week induction regimen followed by six 10-week cycles of maintenance chemotherapy. Feasibility was evaluated with short-term and long-term endpoints. Short-term feasibility was evaluated by the ability to complete induction and 1 maintenance cycle in 24 weeks without >25% reduction in either carboplatin or temozolomide. Long-term feasibility was evaluated by the ability to administer induction and 4 maintenance cycles within 60 weeks without >25% reduction in either carboplatin or temozolomide. Efficacy was assessed by response to initial chemotherapy and 5-year event-free survival. Initial pathology was reviewed centrally. Sixty-six patients were enrolled on the study. It was feasible to deliver the regimen, and toxicity was acceptable. The only significant toxicities were hematologic. Both the short-term and long-term feasibility endpoints were met. The short-term feasibility success rate was 87% (95% CI: 77%-96%) and the long-term feasibility success rate was 79% (95% CI: 68%-90%). The 5-year event-free survival was 46% (95% CI: 33%-58%) and the 5-year survival was 87% (95% CI: 75%-93%). It was feasible to deliver the combination of weekly carboplatin and vincristine alternating with temozolomide to children with progressive/symptomatic low-grade glioma with acceptable toxicities. This combination appears to be effective in delaying progression. Further trials are needed to establish the relative efficacy of this regimen compared with other regimens in use.


The Influence of Online Experiences: The Shaping of Gay Male Identities.
Although research has documented that technological advances via online avenues have impacted the way LGBT individuals gain social capital, less is known about how these online experiences influence the coming out processes for gay men. Using a qualitative approach, this study seeks to understand how the online experiences of gay men either enhanced or inhibited the coming out process. Clustered themes revealed four insights: 1. Online experiences catalyzed the coming out process, 2. Early milestones about the realization of homosexuality came largely from online experiences, 3. Pornography was a tool often utilized for sexual knowledge, and 4. Online experiences helped affirm homosexual identity. Generally, data show that online experiences may manifest offline experiences (i.e., meeting up with another individual, talking about their sexual orientation with others), and that technological communication expedites the coming out process.


[Chinese expert consensus on clinical application of endobiliary radiofrequency ablation therapy for malignant biliary obstruction].
Endobiliary radiofrequency ablation (EB-RFA) is a novel therapy for malignant biliary obstruction (MBO), which is considered as a palliative treatment for MBO caused by cholangiocarcinoma, pancreatic head carcinoma, metastatic lymphadenopathy and other causes. Due to the advantages of minimal invasiveness and repeatability, EB-RFA plays an important role in the combination of stent implantation in the initial treatment of MBO or in the recanalization of stent stenosis/obstruction. In this expert consensus, the indications, contraindications, operation protocol and evaluation criteria of efficacy of EB-RFA were sufficiently discussed, which was organized by the Professional Committee of Minimally Invasive Therapy of China Anti-cancer Association.


Glial cells influence cardiac permittivity as evidenced through in vitro and in silico models.
Excitation-contraction (EC) coupling in the heart has, until recently, been solely accredited to cardiomyocytes. The inherent complexities of the heart make it difficult to examine non-muscle contributions to contraction in vivo, and conventional in vitro models fail to capture multiple features and cellular heterogeneity of the myocardium. Here, we report on the development of a 3D cardiac μTissue to investigate changes in the cellular composition of native myocardium in vitro. Cells are encapsulated within micropatterned gelatin-based hydrogels formed via visible light photocrosslinking. This system enables spatial control of the microarchitecture, perturbation of the cellular composition, and functional measures of EC coupling via video microscopy and a custom algorithm to quantify beat frequency and degree of coordination. To demonstrate the robustness of these tools and evaluate the impact of altered cell population densities on cardiac μTissues, contractility and cell morphology were assessed with the inclusion of exogenous non-myelinating Schwann cells (SCs). Results demonstrate that the addition of exogenous SCs alter cardiomyocyte EC, profoundly inhibiting the response to electrical pacing. Computational modeling of connexin-mediated coupling suggests that SCs impact cardiomyocyte resting potential and rectification following depolarization. Cardiac μTissues hold potential for examining the role of cellular heterogeneity in heart health, pathologies, and cellular therapies.


Lymphodepletional strategies in transplantation.
Because lymphocytes were shown to mediate transplant rejection, their depletion has been studied as a mechanism of preventing rejection and perhaps inducing immunologic tolerance. Agents that profoundly deplete lymphocytes have included monoclonal antibodies, cytotoxic drugs, and radiation. We have studied several such agents but focused on antibodies that deplete not only peripheral blood lymphocytes, but also lymph node lymphocytes. Depletion of lymph node T lymphocytes appears to permit peripheral tolerance at least for T cells in animal models. Nevertheless, B-cell responses may be resistant to such approaches, and T memory cells are likewise relatively resistant to depleting antibodies. We review the experimental and clinical approaches to depletion strategies and outline some of the pitfalls of depletion, such as limitations of currently available agents, duration of tolerance, infection, and malignancy. It is notable that most tolerogenic strategies that have been attempted experimentally and clinically include depleting agents even when they are not named as the underlying strategy. Thus, there is an implicitly acknowledged role for reducing the precursor frequency of donor antigen-specific lymphocytes when approaching the daunting goal of transplant tolerance.


Integrating science and practice in pharmacy curricula.
An integrated curriculum is one where the summation of different academic disciplines forms a coherent whole and, importantly, where the relationships between the different disciplines have been carefully and strategically considered when forming the composite. Within pharmacy curriculum integration is important in order to produce graduates who have the capacity to apply their knowledge to a range of complex problems where available information is often incomplete. This paper discusses the development of an integrated curriculum in which students are presented with an organized, logical sequence of material, but still challenged to make their own integrations and develop as integrative thinkers. An evidence-based model upon which an interdisciplinary undergraduate pharmacy curriculum can be built is presented.


Innate immune recognition of flagellin limits systemic persistence of Brucella.
Brucella are facultative intracellular bacteria that cause chronic infections by limiting innate immune recognition. It is currently unknown whether Brucella FliC flagellin, the monomeric subunit of flagellar filament, is sensed by the host during infection. Here, we used two mutants of Brucella melitensis, either lacking or overexpressing flagellin, to show that FliC hinders bacterial replication in vivo. The use of cells and mice genetically deficient for different components of inflammasomes suggested that FliC was a target of the cytosolic innate immune receptor NLRC4 in vivo but not in macrophages in vitro where the response to FliC was nevertheless dependent on the cytosolic adaptor ASC, therefore suggesting a new pathway of cytosolic flagellin sensing. However, our work also suggested that the lack of TLR5 activity of Brucella flagellin and the regulation of its synthesis and/or delivery into host cells are both part of the stealthy strategy of Brucella towards the innate immune system. Nevertheless, as a flagellin-deficient mutant of B. melitensis wasfound to cause histologically demonstrable injuries in the spleen of infected mice, we suggested that recognition of FliC plays a role in the immunological stand-off between Brucella and its host, which is characterized by a persistent infection with limited inflammatory pathology.


Immunotoxicogenomics: the potential of genomics technology in the immunotoxicity risk assessment process.
Evaluation of xenobiotic-induced changes in gene expression as a method to identify and classify potential toxicants is being pursued by industry and regulatory agencies worldwide. A workshop was held at the Research Triangle Park campus of the Environmental Protection Agency to discuss the current state-of-the-science of "immunotoxicogenomics" and to explore the potential role of genomics techniques for immunotoxicity testing. The genesis of the workshop was the current lack of widely accepted triggering criteria for Tier 1 immunotoxicity testing in the context of routine toxicity testing data, the realization that traditional screening methods would require an inordinate number of animals and are inadequate to handle the number of chemicals that may need to be screened (e.g., high production volume compounds) and the absence of an organized effort to address the state-of-the-science of toxicogenomics in the identification of immunotoxic compounds. The major focus of the meeting was on the theoretical and practical utility of genomics techniques to (1) replace or supplement current immunotoxicity screening procedures, (2) provide insight into potential modes or mechanisms of action, and (3) provide data suitable for immunotoxicity hazard identification or risk assessment. The latter goal is of considerable interest to a variety of stakeholders as a means to reduce animal use and to decrease the cost of conducting and interpreting standard toxicity tests. A number of data gaps were identified that included a lack of dose response and kinetic data for known immunotoxic compounds and a general lack of data correlating genomic alterations to functional changes observed in vivo. Participants concluded that a genomics approach to screen chemicals for immunotoxic potential or to generate data useful to risk assessors holds promise but that routine use of these methods is years in the future. However, recent progress in molecular immunology has made mode and mechanism of action studies much more practical. Furthermore, a variety of published immunotoxicity studies suggest that microarray analysis is already a practical means to explore pathway-level changes that lead to altered immune function. To help move the science of immunotoxicogenomics forward, a partnership of industry, academia, and government was suggested to address data gaps, validation, quality assurance, and protocol development.


The heritability of inducible defenses in tadpoles.
The evolution of plastic traits requires phenotypic trade-offs and heritable traits, yet the latter requirement has received little attention, especially for predator-induced traits. Using a half-sib design, I examined the narrow-sense heritability of predator-induced behaviour, morphology, and life history in larval wood frogs (Rana sylvatica). Many of the traits had significant additive genetic variation in predator (caged Anax longipes) and no-predator environments. Whereas most traits had moderate to high heritability across environments, tail depth exhibited high heritability with predators but low heritability without predators. In addition, several traits had significant heritability for plasticity, suggesting a potential for selection to act on plasticity per se. Genetic correlations confirmed known phenotypic relationships across environments and identified novel relationships within each environment. This appears to be the first investigation of narrow-sense heritabilities for predator-induced traits and confirms that inducible traits previously shown to be under selection also have a genetic basis and should be capable of exhibiting evolutionary responses.


Multiple sclerosis: changes in microarchitecture of white matter tracts after training with a video game balance board.
To determine if high-intensity, task-oriented, visual feedback training with a video game balance board (Nintendo Wii) induces significant changes in diffusion-tensor imaging ( DTI diffusion-tensor imaging ) parameters of cerebellar connections and other supratentorial associative bundles and if these changes are related to clinical improvement in patients with multiple sclerosis. The protocol was approved by local ethical committee; each participant provided written informed consent. In this 24-week, randomized, two-period crossover pilot study, 27 patients underwent static posturography and brain magnetic resonance (MR) imaging at study entry, after the first 12-week period, and at study termination. Thirteen patients started a 12-week training program followed by a 12-week period without any intervention, while 14 patients received the intervention in reverse order. Fifteen healthy subjects also underwent MR imaging once and underwent static posturography. Virtual dissection of white matter tracts was performed with streamline tractography; values of DTI diffusion-tensor imaging parameters were then obtained for each dissected tract. Repeated measures analyses of variance were performed to evaluate whether DTI diffusion-tensor imaging parameters significantly changed after intervention, with false discovery rate correction for multiple hypothesis testing. There were relevant differences between patients and healthy control subjects in postural sway and DTI diffusion-tensor imaging parameters (P < .05). Significant main effects of time by group interaction for fractional anisotropy and radial diffusivity of the left and right superior cerebellar peduncles were found (F2,23 range, 5.555-3.450; P = .036-.088 after false discovery rate correction). These changes correlated with objective measures of balance improvement detected at static posturography (r = -0.381 to 0.401, P < .05). However, both clinical and DTI diffusion-tensor imaging changes did not persist beyond 12 weeks after training. Despite the low statistical power (35%) due to the small sample size, the results showed that training with the balance board system modified the microstructure of superior cerebellar peduncles. The clinical improvement observed after training might be mediated by enhanced myelination-related processes, suggesting that high-intensity, task-oriented exercises could induce favorable microstructural changes in the brains of patients with multiple sclerosis.


A crystallization-induced stereoselective glycosidation reaction in the synthesis of the anticancer drug etoposide.
The anticancer drug etoposide, 1, is prepared in 79% overall yield from readily available 4'-demethyl-4-epipodophyllotoxin, 3, and 4, 6-O-ethylidene-2,3-O-dibenzyl-D-glucose, 4, via a crystallization-induced stereoselective glycosidation reaction followed by catalytic hydrogenation.


Differential expression and tumor necrosis factor-mediated regulation of TNFRSF11b/osteoprotegerin production by human melanomas.
Tumors escape host immune responses, in part, through the release of immunomodulatory factors and decoy receptors into their microenvironment. Several cancers express surface-bound and soluble members of the tumor necrosis factor (TNF) receptor superfamily, including TNFRSF11b/osteoprotegerin (OPG). In its physiologic role, OPG regulates bone remodeling through competition for osteoclast-activating cytokines and protects newly forming bone from T cell-mediated apoptosis. In multiple tumor types, OPG production is associated with an aggressive phenotype and increased metastasis to bone, but no study has examined OPG production in human metastatic melanoma. We demonstrate that a significant proportion of human metastatic melanomas constitutively produces OPG through a mechanism governed by membrane-bound TNF-α signaling through TNF receptor 1 (TNFR1). These observations both define a specific mechanism that regulates melanoma production of OPG and establish a new molecular target for the therapeutic regulation of OPG.


A multilevel analysis of socioeconomic status and prostate cancer risk.
We investigated whether prostate cancer was associated with socioeconomic status (SES) at the individual level, area level, or a combination of both levels. This population-based case-control study of prostate cancer in men aged 65 to 79 years was conducted between 2000 and 2002 in South Carolina. Complete interviews were available for 407 incident prostate cancer cases and 393 controls (with respective response rates of 61% and 64%). We used educational level to measure individual-level SES and a composite variable capturing income and education from 2000 Census data to measure area-level SES. After adjustment for race, age, geographic region, and prostate-specific antigen testing, men with some college were at reduced risk for prostate cancer (odds ratio [OR], 0.44; 95% confidence interval [CI], 0.27-0.72), as were men in the highest quartile of area-level SES (OR, 0.52; 95% CI, 0.34-0.80). When assessing individual-level and area-level SES simultaneously and accounting for their nonindependence, the independent negative associations persisted and appeared to be more striking for men with a diagnosis of localized disease, rather than advanced disease. The independent effects of area-level and individual-level SES on prostate cancer risk seen in our study may help explain the conflicting results of previous studies conducted at both levels.


Using ancestral information to detect and localize quantitative trait loci in genome-wide association studies.
In mammalian genetics, many quantitative traits, such as blood pressure, are thought to be influenced by specific genes, but are also affected by environmental factors, making the associated genes difficult to identify and locate from genetic data alone. In particular, the application of classical statistical methods to single nucleotide polymorphism (SNP) data collected in genome-wide association studies has been especially challenging. We propose a coalescent approach to search for SNPs associated with quantitative traits in genome-wide association study (GWAS) data by taking into account the evolutionary history among SNPs. We evaluate the performance of the new method using simulated data, and find that it performs at least as well as existing methods with an increase in performance in the case of population structure. Application of the methodology to a real data set consisting of high-density lipoprotein cholesterol measurements in mice shows the method performs well for empirical data, as well. By combining methods from stochastic processes and phylogenetics, this work provides an innovative avenue for the development of new statistical methodology in the analysis of GWAS data.


Antibiotics for acute otitis media in children.
Acute otitis media is one of the most common diseases in early infancy and childhood. Antibiotic use for acute otitis media varies from 31% in the Netherlands to 98% in the USA and Australia. The objective of this review was to assess the effects of antibiotics for children with acute otitis media. We searched the Cochrane Controlled Trials Register, MEDLINE, Index Medicus (pre 1965), Current Contents and reference lists of articles from 1958 to January 2000. Randomised trials comparing antimicrobial drugs with placebo in children with acute otitis media. Three reviewers independently assessed trial quality and extracted data. Ten trials were eligible but only seven trials, with a total of 2,202 children, included patient-relevant outcomes. The methodological quality of the included trials was generally high. All trials were from developed countries. The trials showed no reduction in pain at 24 hours, but a 28% relative reduction (95% confidence interval 15% to 38%) in pain at two to seven days. Since approximately 80% of patients will have settled spontaneously in this time, this means an absolute reduction of 5% or that about 17 children must be treated with antibiotics to prevent one child having some pain after two days. There was no effect of antibiotics on hearing problems of acute otitis media, as measured by subsequent tympanometry. However, audiometry was done in only two studies and incompletely reported. Nor did antibiotics influence other complications or recurrence. There were few serious complications seen in these trials: only one case of mastoiditis occurred in a penicillin treated group. Antibiotics provide a small benefit for acute otitis media in children. As most cases will resolve spontaneously, this benefit must be weighed against the possible adverse reactions. Antibiotic treatment may play an important role in reducing the risk of mastoiditis in populations where it is more common. [This abstract has been prepared centrally.]


An Evaluation of Entropy Measures for Microphone Identification.
Research findings have shown that microphones can be uniquely identified by audio recordings since physical features of the microphone components leave repeatable and distinguishable traces on the audio stream. This property can be exploited in security applications to perform the identification of a mobile phone through the built-in microphone. The problem is to determine an accurate but also efficient representation of the physical characteristics, which is not known a priori. Usually there is a trade-off between the identification accuracy and the time requested to perform the classification. Various approaches have been used in literature to deal with it, ranging from the application of handcrafted statistical features to the recent application of deep learning techniques. This paper evaluates the application of different entropy measures (Shannon Entropy, Permutation Entropy, Dispersion Entropy, Approximate Entropy, Sample Entropy, and Fuzzy Entropy) and their suitability for microphone classification. The analysis is validated against an experimental dataset of built-in microphones of 34 mobile phones, stimulated by three different audio signals. The findings show that selected entropy measures can provide a very high identification accuracy in comparison to other statistical features and that they can be robust against the presence of noise. This paper performs an extensive analysis based on filter features selection methods to identify the most discriminating entropy measures and the related hyper-parameters (e.g., embedding dimension). Results on the trade-off between accuracy and classification time are also presented.


Trends in resistant Enterobacteriaceae and Acinetobacter species in hospitalized patients in the United States: 2013-2017.
Trends in antimicrobial resistance help inform infection control efforts. We examined trends in resistance for Enterobacteriaceae and Acinetobacter spp. from 2013 to 2017 in hospitalized US patients. We analyzed antimicrobial susceptibility of non-duplicate isolates in hospitalized patients (not limited to hospital-acquired infections) in the US BD Insights Research Database. Resistance profiles of interest were extended-spectrum beta-lactamase (ESBL)-producing, multidrug resistant (MDR), and carbapenem-nonsusceptible (Carb-NS) phenotypes of Enterobacteriaceae, and MDR and Carb-NS Acinetobacter spp. Time series models were used to evaluate the patterns of resistance trends in rate per 100 hospital admissions and proportion per isolates tested. More than 1 million Enterobacteriaceae isolates were obtained from 411 hospitals; 12.05% were ESBL, 1.21% Carb-NS, and 7.08% MDR. Urine was the most common source. For Acinetobacter spp. (n = 19,325), 37.48% were Carb-NS, 47.66% were MDR, and the most common source was skin/wound cultures. Trend analyses showed that the rates of ESBL and Carb-NS Enterobacteriaceae per 100 hospital admissions increased significantly between 2013 and 2017. Rates of MDR Enterobacteriaceae and Carb-NS and MDR Acinetobacter spp. decreased during this time period. Trends in proportions of resistant isolates generally mirrored trends in rates per 100 hospital admissions. MDR Enterobacteriaceae and Carb-NS and MDR Acinetobacter spp. were more common in winter than summer. In this large-scale study of patients in US hospitals, rates of ESBL and Carb-NS Enterobacteriaceae per 100 hospital admissions increased between 2013 and 2017. MDR Enterobacteriaceae and MDR and Carb-NS Acinetobacter spp. isolates decreased over this period. These data support continuing infection control and stewardship efforts and the development of new therapeutic options.


Infundibular dilations of the posterior communicating arteries: pathogenesis, anatomical variants, aneurysm formation, and subarachnoid hemorrhage.
Cerebrovascular infundibular dilations (IDs) are triangular-shaped widenings less than 3 mm in diameter, which are most commonly found at the posterior communicating artery (PCoA). The aims of this systematic review are to elucidate the natural histories of IDs, determine their risk of progression to significant pathology, and discuss potential management options. A comprehensive literature search of PubMed was used to find all case reports and series relating to cerebral IDs. IDs were classified into three types: type I IDs do not exhibit morphological change over a long follow-up period, type II IDs evolve into saccular aneurysms, while type III IDs are those that result in subarachnoid hemorrhage without prior aneurysmal progression. Data were extracted from studies that demonstrated type II or III IDs. We reviewed 16 cases of type II and seven cases of type III IDs. For type II IDs, 81.3% of patients were female with a median age at diagnosis of 38. All type II IDs were located at the PCoA without a clear predilection for sidedness. Median time to aneurysm progression was 7.5 years. For type III IDs there was no clear gender preponderance and the median age at diagnosis was 51. The PCoA was involved in 85.7% of cases, with 57.1% of IDs occurring on the left. Most patients were treated with clipping. Risk factors for aneurysm formation appear to be female gender, young age, left-sided localization, coexisting aneurysms, and hypertension. IDs can rarely progress to aneurysms or rupture. Young patients with type II or III IDs with coexisting aneurysms or hypertension may benefit from long-term imaging surveillance.


Quercetin sensitizes cells in a tumour-like low pH environment to hyperthermia.
Quercetin has been shown to act as a hyperthermia sensitizer by inhibiting the synthesis of heat shock protein 70 (HSP70) in a variety of tumour cell lines. It is most effective under conditions of low pH. This study was designed to test the hypothesis that quercetin suppresses thermotolerance development in cells adapted to growth at low pH and renders them as responsive as acutely acidified cells to hyperthermia-induced cytotoxicity. Chinese hamster ovarian carcinoma cells (OvCa) were exposed to 42 degrees C hyperthermia and/or quercetin (50-200 mm) at their growth pH of either 7.3 or 6.7 or after acute acidification from 7.3 to 6.7. Thermotolerance development was measured by colony survival. HSP70 synthesis and total protein synthesis were measured by radioactive precursor pulse labelling techniques. Quercetin, in a concentration-dependent manner, reduced the rate of total protein synthesis and increased cytotoxicity equally after acute acidification to pH 6.7 or growth at pH 6.7 at 37 degrees C, and to a greater extent than it did in cells at pH 7.3. At 42 degrees C, 100 mm quercetin inhibited total protein synthesis, HSP70 synthesis and thermotolerance development to a similar extent in cells grown at pH 6.7 or acutely acidified to pH 6.7. In contrast, quercetin reduced but did not completely inhibit HSP70 synthesis and thermotolerance development in cells grown and heated at pH 7.3. These results support the hypothesis that quercetin can specifically reduce thermotolerance development in tumour cells adapted to growth at pHe 6.7 so that they respond similarly to acutely acidified cells. Since many tumours are adapted to growth at low pH and may resist a wide variety of therapeutic modalities, inhibition of thermotolerance expression by quercetin may not only enhance the response to hyperthermia but the response to commonly used therapies such as chemotherapy and radiation.


An artificial neural network (ANN)-based lung-tumor motion predictor for intrafractional MR tumor tracking.
To address practical issues of implementing artificial neural networks (ANN) for lung-tumor motion prediction in MRI-based intrafractional lung-tumor tracking. A feedforward four-layered ANN structure is used to predict future tumor positions. A back-propagation algorithm is used for ANN learning. Adaptive learning is incorporated by continuously updating weights and learning rate during prediction. An ANN training scheme specific for MRI-based tracking is developed. A multiple-ANN structure is developed to reduce tracking failures caused by the lower imaging rates of MRI. We used particle swarm optimization to optimize the ANN structure and initial weights (IW) for each patient and treatment fraction. Prediction accuracy is evaluated using the 1D superior-inferior lung-tumor motions of 29 lung cancer patients for system delays of 120-520 ms, in increments of 80 ms. The result is compared with four different scenarios: (1), (2) ANN structure optimization + with∕without IW optimization, and (3), (4) no ANN structure optimization + with∕without IW optimization, respectively. An additional simulation is performed to assess the value of optimizing the ANN structure for each treatment fraction. For 120-520 ms system delays, mean RMSE values (ranges 0.0-2.8 mm from 29 patients) of 0.5-0.9 mm are observed, respectively. Using patient specific ANN structures, a 30%-60% decrease in mean RMSE values is observed as a result of IW optimization, alone. No significant advantages in prediction performance are observed, however, by optimizing for each fraction. A new ANN-based lung-tumor motion predictor is developed for MRI-based intrafractional tumor tracking. The prediction accuracy of our predictor is evaluated using a realistic simulated MR imaging rate and system delays. For 120-520 ms system delays, mean RMSE values of 0.5-0.9 mm (ranges 0.0-2.8 mm from 29 patients) are achieved. Further, the advantage of patient specific ANN structure and IW in lung-tumor motion prediction is demonstrated by a 30%-60% decrease in mean RMSE values.


Vancomycin use in a large teaching hospital in Shiraz, Islamic Republic of Iran, 2003.
We investigated adherence to the Hospital Infection Control Practice Advisory Committee (HICPAC) guidelines on vancomycin prescription in a large university-affiliated hospital in Shiraz. From August to December 2003, 200 hospitalized patients received vancomycin. For only 12 (6%) of these patients was vancomycin prescribed appropriately according to HICPAC guidelines. The main reasons why vancomycin use did not comply with HICPAC recommendations were: surgical prophylaxis in patients with negative cultures for resistant Gram-positive organisms, no investigation of vancomycin serum levels in patients receiving > 48 hours of vancomycin, vancomycin serum levels not repeated in patients receiving > 1 week of vancomycin, no appropriate adjustment of dosage with respect to serum levels in patients receiving vancomycin.


The checkered history of American psychiatric epidemiology.
American psychiatry has been fascinated with statistics ever since the specialty was created in the early nineteenth century. Initially, psychiatrists hoped that statistics would reveal the benefits of institutional care. Nevertheless, their fascination with statistics was far removed from the growing importance of epidemiology generally. The impetus to create an epidemiology of mental disorders came from the emerging social sciences, whose members were concerned with developing a scientific understanding of individual and social behavior and applying it to a series of pressing social problems. Beginning in the 1920s, the interest of psychiatric epidemiologists shifted to the ways that social environments contributed to the development of mental disorders. This emphasis dramatically changed after 1980 when the policy focus of psychiatric epidemiology became the early identification and prevention of mental illness in individuals. This article reviews the major developments in psychiatric epidemiology over the past century and a half. The lack of an adequate classification system for mental illness has precluded the field of psychiatric epidemiology from providing causal understandings that could contribute to more adequate policies to remediate psychiatric disorders. Because of this gap, the policy influence of psychiatric epidemiology has stemmed more from institutional and ideological concerns than from knowledge about the causes of mental disorders. Most of the problems that have bedeviled psychiatric epidemiology since its inception remain unresolved. In particular, until epidemiologists develop adequate methods to measure mental illnesses in community populations, the policy contributions of this field will not be fully realized.


Maternal thyroid autoimmunity during pregnancy and the risk of attention deficit/hyperactivity problems in children: the Generation R Study.
Maternal thyroid status and autoimmunity during pregnancy have been associated with impaired development of the offspring in animal and human studies. Our objective was to examine whether elevated titers of maternal thyroid peroxidase antibodies (TPOAbs) in early pregnancy increased the risk of cognitive impairment and problem behavior in preschool children. Second, we aimed at exploring to what extent any effect on child behavior was mediated by maternal thyroid parameters during pregnancy. In the Generation R Study, a population-based cohort of 3139 children and their mothers, we measured maternal thyroid parameters (thyrotropin [TSH], free Thyroxine, and TPOAbs) at 13.5±1.8 weeks of gestation. Children's verbal and nonverbal cognitive functioning was measured at 2.5 years using the Language Development Survey and the Parent Report of Children Abilities. At 3 years, children's behavior was assessed using the Child Behavior Checklist. Elevated titers of TPOAbs during pregnancy did not predict the verbal and nonverbal cognitive functioning of the children. However, elevated titers of TPOAbs in mothers were associated with externalizing problems in children (odds ratio [OR]=1.64, 95% confidence interval [CI]: 1.17-2.29, p=0.004). In particular, children of TPOAb-positive mothers were at a higher risk of attention deficit/hyperactivity problems (OR=1.77, 95% CI: 1.15-2.72, p=0.01). To explore whether the effect of maternal TPOAbs on child problem behavior was mediated by maternal thyroid parameters, we added maternal TSH to the model. After correcting for TSH, the effect of TPOAbs on externalizing problems was attenuated slightly but remained significant (OR=1.56, 95% CI: 1.14, 2.14, p=0.005). Our findings imply that the elevated titers of TPOAbs during pregnancy impact children's risk of problem behavior, in particular, attention deficit/hyperactivity. The observed effect is only partially explained by maternal TSH levels. These findings may point to a specific mechanism of Attention Deficit/Hyperactivity Disorder in children. Nevertheless, we can only speculate about public health implication of the study, as there is no specific treatment for TPOAb-positive pregnant women with normal thyroid function. Further investigation is needed to explore whether TPOAb-positive pregnant women and their children can benefit from close monitoring and early detection of developmental delay in populations at risk.


Visual imagery deficits, impaired strategic retrieval, or memory loss: disentangling the nature of an amnesic person's autobiographical memory deficit.
Conclusions about the duration of hippocampal contributions to our autobiographical record of personal episodes have come under intense scrutiny in recent years. Interpretation is complicated by such factors as extent and site of lesions as well as test sensitivity. We describe the case of an amnesic person, K.C., with large, bilateral hippocampal lesions who figured prominently in the development of theories of remote memory due to his severely impoverished autobiographical memory extending across his entire lifetime. However, the presence of lesions in higher-order visual cortex raises the possibility that K.C.'s retrograde autobiographical amnesia is mediated by loss of long-term visual images, whereas widespread frontal lesions suggest that his impairment may relate to deficits in strategic retrieval rather than storage. Normal performance on an extensive battery of visual imagery tests refutes the imagery loss interpretation. To test for deficits in strategic retrieval, we used a more formal autobiographical memory test requiring generation of personal events under varying levels of retrieval support. However, even with rigorous contextual prompting, K.C. produced few pre-injury recollections; all were schematic, lacking the richness of detail produced by control participants, raising doubt that his deficit is one of retrieval. Findings are discussed in the context of theories concerning the duration of hippocampal-neocortical interactions in supporting autobiographical re-experiencing. The approach we used to investigate the effects of different lesions on memory provides a framework for dealing with other patients who present with an interesting functional deficit whose neuroanatomical source is difficult to specify due to widespread lesions.


Basin stability measure of different steady states in coupled oscillators.
In this report, we investigate the stabilization of saddle fixed points in coupled oscillators where individual oscillators exhibit the saddle fixed points. The coupled oscillators may have two structurally different types of suppressed states, namely amplitude death and oscillation death. The stabilization of saddle equilibrium point refers to the amplitude death state where oscillations are ceased and all the oscillators converge to the single stable steady state via inverse pitchfork bifurcation. Due to multistability features of oscillation death states, linear stability theory fails to analyze the stability of such states analytically, so we quantify all the states by basin stability measurement which is an universal nonlocal nonlinear concept and it interplays with the volume of basins of attractions. We also observe multi-clustered oscillation death states in a random network and measure them using basin stability framework. To explore such phenomena we choose a network of coupled Duffing-Holmes and Lorenz oscillators which are interacting through mean-field coupling. We investigate how basin stability for different steady states depends on mean-field density and coupling strength. We also analytically derive stability conditions for different steady states and confirm by rigorous bifurcation analysis.


Sulforaphane produces antidepressant- and anxiolytic-like effects in adult mice.
Increasing evidence suggests that depression is accompanied by dysregulation of neuroimmune system. Sulforaphane (SFN) is a natural compound with antioxidative, anti-inflammatory and neuroprotective activities. The present study aims to investigate the effects of SFN on depressive- and anxiety-like behaviors as well as potential neuroimmune mechanisms in mice. Repeated SFN administration (10mg/kg, i.p.) significantly decreased the immobility time in the forced swimming test (FST), tail suspension test (TST), and latency time to feeding in the novelty suppressed feeding test (NSF), and increased the time in the central zone in the open field test (OPT). Using the chronic mild stress (CMS) paradigm, we confirmed that repeated SFN (10mg/kg, i.p.) administration significantly increased sucrose preference in the sucrose preference test (SPT), and immobility time in the FST and TST of mice subjected to CMS. Also, SFN treatment significantly reversed anxiety-like behaviors (assessed by the OPT and NSF) of chronically stressed mice. Finally, ELISA analysis showed that SFN administration blocked the increase in the serum levels of corticosterone (CORT), adrenocorticotropic hormone (ACTH), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in chronically stressed mice. In summary, these findings demonstrated that SFN has antidepressant- and anxiolytic-like activities in stressed mice model of depression, which likely occurs by inhibiting the hypothalamic-pituitary-adrenal (HPA) axis and inflammatory response to stress. These data support further exploration for developing SFN as a novel agent to treat depression and anxiety disorders.


Muscle contributions to support and progression during single-limb stance in crouch gait.
Pathological movement patterns like crouch gait are characterized by abnormal kinematics and muscle activations that alter how muscles support the body weight during walking. Individual muscles are often the target of interventions to improve crouch gait, yet the roles of individual muscles during crouch gait remain unknown. The goal of this study was to examine how muscles contribute to mass center accelerations and joint angular accelerations during single-limb stance in crouch gait, and compare these contributions to unimpaired gait. Subject-specific dynamic simulations were created for ten children who walked in a mild crouch gait and had no previous surgeries. The simulations were analyzed to determine the acceleration of the mass center and angular accelerations of the hip, knee, and ankle generated by individual muscles. The results of this analysis indicate that children walking in crouch gait have less passive skeletal support of body weight and utilize substantially higher muscle forces to walk than unimpaired individuals. Crouch gait relies on the same muscles as unimpaired gait to accelerate the mass center upward, including the soleus, vasti, gastrocnemius, gluteus medius, rectus femoris, and gluteus maximus. However, during crouch gait, these muscles are active throughout single-limb stance, in contrast to the modulation of muscle forces seen during single-limb stance in an unimpaired gait. Subjects walking in crouch gait rely more on proximal muscles, including the gluteus medius and hamstrings, to accelerate the mass center forward during single-limb stance than subjects with an unimpaired gait.


The Lhx4 homeobox transcript in the rat pineal gland: Adrenergic regulation and impact on transcripts encoding melatonin-synthesizing enzymes.
Homeobox genes generally encode transcription factors involved in regulating developmental processes. In the pineal gland, a brain structure devoted to nocturnal melatonin synthesis, a number of homeobox genes are also expressed postnatally; among these is the LIM homeobox 4 gene (Lhx4). We here report that Lhx4 is specifically expressed in the postnatal pineal gland of rats and humans. Circadian analyses revealed a fourfold rhythm in Lhx4 expression in the rat pineal gland, with rhythmic expression detectable from postnatal day 10. Pineal Lhx4 expression was confirmed to be positively driven by adrenergic signaling, as evidenced by in vivo modulation of Lhx4 expression by pharmacological (isoprenaline injection) and surgical (superior cervical ganglionectomy) interventions. In cultured pinealocytes, Lhx4 expression was upregulated by cyclic AMP, a second messenger of norepinephrine. By use of RNAscope technology, Lhx4 transcripts were found to be exclusively localized in melatonin-synthesizing pinealocytes. This prompted us to investigate the possible role of Lhx4 in regulation of melatonin-producing enzymes. By use of siRNA technology, we knocked down Lhx4 by 95% in cultured pinealocytes; this caused a reduction in transcripts encoding the melatonin-producing enzyme arylalkylamine N-acetyl transferase (Aanat). Screening the transcriptome of siRNA-treated pinealocytes by RNAseq revealed a significant impact of Lhx4 on the phototransduction pathway and on transcripts involved in development of the nervous system and photoreceptors. These data suggest that rhythmic expression of Lhx4 in the pineal gland is controlled via an adrenergic-cyclic AMP mechanism and that Lhx4 acts to promote nocturnal melatonin synthesis.


Tests for occult in stools of children.
Three tests for the presence of occult blood in the stools of children were examined; the orthotolidine test, the guaiacum resin test, and a modified reduced phenolphthalain test. The stools of 71 neonates, 33 normal children, and 8 children with suspected gastrointestinal pathology were examined. The 1% orthotolidine test and the guaiacum resin test produced a high number of false positive results in healthy children eating a meat-containing diet. It is concluded that a positive result by these tests is of little diagnostic value in children eating a normal diet.


Anti-angiogenic gene therapy for hepatocellular carcinoma mediated by microbubble-enhanced ultrasound exposure: an in vivo experimental study.
The aim of this study was to assess the effect of anti-angiogenic gene therapy for hepatocelluar carcinoma (HCC) treated by microbubble-enhanced ultrasound exposure. Forty C57BL/6J female mice were inoculated s.c. with Hepa1-6 tumor cell line. Herpes simplex virus thymidine kinase under the control of kinase domain-containing receptor (KDR, angiogenic growth factor's corresponding receptor) promoter was used. Plasmid DNA with or without microbubble contrast agent of SonoVuetrade mark was i.v. injected. Ultrasound (1 MHz, 2 W/cm(2), 5 min) was delivered to hepatic carcinomas in mice. The KDR-tk gene transfer was followed by ganciclovir (GCV) injection for 10 days and then the diameters of tumors were measured every 4 days till 28 days. The survivals of tumor-bearing mice were observed. PCR analysis and immunohistochemistry measurements revealed expression of the transfected gene. Transferase-mediated dUTP nick end labeling staining was used to detect apoptotic cells. Compared with the group treated by ultrasound alone, KDR-tk gene treatment treated by ultrasound combined with SonoVue restrained tumor growth and increased survival time of tumor-bearing mice; microvessel density in group mediated by ultrasound and SonoVue was significantly lower than that in group ultrasound alone (12.3 +/- 1.4 vs. 27.4 +/- 3.2, P < 0.05). An apoptosis index increased in the group treated by ultrasound and SonoVue compared with the group treated by ultrasound alone (25 +/- 3.6 vs. 36 +/- 3.8, P < 0.05), whereas there was no significant difference between group mediated by SonoVue alone and group phosphate-buffered saline alone (17 +/- 1.8 vs. 14 +/- 1.2, P>0.05). Gene therapy mediated by ultrasound exposure enhanced by a microbubble contrast agent may become a new treatment option for persistent HCC.


Helpfulness, Trust, and Safety of Neighborhoods: Social Capital, Household Income, and Self-Reported Health of Older Adults.
Growing literature documents that where you live has an impact on your health, due in part to social capital. Building on social capital literature, we assess how subjective appraisals of neighborhood quality are associated with self-reported health (SRH) for older adults. Cross-sectional analysis of the 2014 California Health Interview Survey, a representative survey of diverse, noninstitutionalized California residents. We use three measures of neighborhood quality: trustworthy neighbors, helpful neighbors, and feeling safe. Using weighted ordinary least squares regression, we assess the associations of trust, helpfulness, and safety to SRH, controlling for neighborhood, demographic, and health care variables. We then examine how these associations vary by household income. We find that characterizing neighbors as helpful and feeling safe are associated with better SRH, even controlling for community, demographic, and health care variables. However, the importance of these dimensions varies across household income: helpfulness is positively associated, whereas trust is negatively associated with SRH for lower income residents; safety is positively associated with SRH in all but the lowest income residents. These findings show that social capital dimensions work differently from one another, and differentially affect the health of older adults. Scholarly analyses of neighborhood effects should include a range of social capital measures and stratify by household income. Our findings may also inform priority setting for social capital programs, especially for older adults with limited economic resources. Policies and programs should consider actions that raise perceptions of helpfulness and safety.


Prey-switching does not protect a generalist turtle from bioenergetic consequences when its preferred food is scarce.
Optimal foraging theory explains how animals make foraging decisions based on the availability, nutritional content, and handling times of different food types. Generalists solve this problem by consuming a variety of food types, and alter their diets with relative ease. Specialists eat few food types, and may starve if those food types are not available. We integrated stable isotope analyses with previously-published stomach contents and environmental data to investigate how the foraging ecologies of three sympatric freshwater turtle species vary across four wetlands that differ in turbidity and primary producer abundance. We found that the generalist Emydura macquarii consumes a varied diet (but mostly filamentous green algae) when primary producers are available and water is clear, but switches to a more carnivorous diet when the water is turbid and primary producers are scarce, following the predictions of optimal foraging theory. In contrast, two more-specialized carnivorous species, Chelodina expansa and Chelodina longicollis, do not differ in diet across wetlands, and interspecific competition may increase where E. macquarii is carnivorous. When forced to be more carnivorous, E. macquarii exhibits higher rates of empty stomachs, and female turtles have reduced body condition, but neither Chelodina species are affected. Our results provide support for optimal foraging theory, but also show that the ability to change diet does not protect the generalist from experiencing lower foraging success when its preferred food is rare, with direct consequences for their energy budgets. Our results have conservation implications because wetlands in the Murray-Darling river system are increasingly turbid and have low macrophyte abundance, and all three species are declining.


Glutathione redox cycle regulates nitric oxide-mediated glyceraldehyde-3-phosphate dehydrogenase inhibition.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been identified as a potential target for nitric oxide (NO)-mediated cellular toxicity. We have previously shown that NO inhibits GAPDH by S-nitrosylation of the active site cysteine residue, which is reversed by low-molecular-weight thiols. Because endothelial cells contain high concentrations of low-molecular-weight thiols, principally glutathione, we investigated the effect of NO on GAPDH activity in intact endothelial cells and the influence that cellular glutathione has on GAPDH inhibition. Our results show that incubation of cells with an exogenous NO-generating system resulted in inhibition of GAPDH activity. The mechanism for inhibition appears to involve reversible modification of GAPDH because addition of thiols to cell extracts restored activity. Furthermore, cells were able to completely recover GAPDH activity after removal of the NO-generating system. Recovery did not require de novo protein synthesis. Depletion of cellular glutathione levels by treatment of cells with buthionine sulfoximine resulted in greater NO-mediated GAPDH inhibition as well as a lesser ability to recover activity. Finally, disruption of the glutathione redox cycle with the glutathione reductase inhibitor, 1,3-bis(2-chloroethyl)-1-nitrosourea, increased the extent of NO-mediated GAPDH inhibition and decreased both the rate and degree of recovery of GAPDH-activity. These results suggest that the glutathione redox cycle plays an important role not only in regulating the extent of NO-mediated GAPDH inhibition but also in the ability of endothelial cells to recover from NO-mediated GAPDH inhibition.


Nanoporous thin films in optical waveguide spectroscopy for chemical analytics.
Spectroscopy with planar optical waveguides is still an active field of research for the quantitative analysis of various supramolecular surface architectures and processes, and for applications in integrated optical chip communication, direct chemical sensing, etc. In this contribution, we summarize some recent development in optical waveguide spectroscopy using nanoporous thin films as the planar substrates that can guide the light just as well as bulk thin films. This is because the nanoporosity is at a spacial length-scale that is far below the wavelength of the guided light; hence, it does not lead to an enhanced scattering or additional losses of the optical guided modes. The pores have mainly two effects: they generate an enormous inner surface (up to a factor of 100 higher than the mere geometric dimensions of the planar substrate) and they allow for the exchange of material and charges between the two sides of the solid thin film. We demonstrate this for several different scenarios including anodized aluminum oxide layers for the ultrasensitive determination of the refractive index of fluids, or the label-free detection of small analytes binding from the pore inner volume to receptors immobilized on the pore surface. Using a thin film of Ti metal for the anodization results in a nanotube array offering an even further enhanced inner surface and the possibility to apply electrical potentials via the resulting TiO2 semiconducting waveguide structure. Nanoporous substrates fabricated from SiNx thin films by colloid lithography, or made from SiO2 by e-beam lithography, will be presented as examples where the porosity is used to allow for the passage of ions in the case of tethered lipid bilayer membranes fused on top of the light-guiding layer, or the transport of protons through membranes used in fuel cell applications. The final example that we present concerns the replication of the nanopore structure by polymers in a process that leads to a nanorod array that is equally well suited to guide the light as the mold; however, it opens a totally new field for integrated optics formats for direct chemical and biomedical sensing with an extension to even molecularly imprinted structures. Graphical abstract.


Making words count: the value of qualitative research.
In the current climate of evidence-based practice, physiotherapy is urged to prove its worth via rigorous scientific research. However, there are concerns that limited methodologies are used to explore complex therapeutic issues, and that the profession relies too heavily on quantitative research studies to provide its evidence base. Qualitative research methods are able to explore the complexity of human behaviour and generate deeper understanding of illness behaviours and therapeutic interactions. Nevertheless, there is still a sense of distrust of qualitative research, related to the challenge of evaluating both the quality and usefulness of findings derived through qualitative methods. This discussion paper explores these issues. It examines some of the most frequently used techniques aimed at ensuring quality and value in qualitative research, such as sampling, triangulation, multiple coding, respondent validation and the use of audit trails, as well as addressing reflexivity. Because of the pluralistic and interactive nature of qualitative inquiry, the criteria used to judge quality need to be appropriate to each piece of research and should provide evidence to help readers to evaluate the calibre of the study and its relevance to their own area of work.


Asymmetric distribution of cytochrome P-450 and NADPH--cytochrome P-450 (cytochrome c) reductase in vesicles from smooth endoplasmic reticulum of rat liver.
1. The topography of cytochrome P-450 in vesicles from smooth endoplasmic reticulum of rat liver has been examined. Approx. 50% of the cytochrome is directly accessible to the action of trypsin in intact vesicles whereas the remainder is inaccessible and partitioned between luminal-facing or phospholipid-embedded loci. Analysis by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis reveals three major species of the cytochrome. Of these, the variant with a mol.wt. of 52000 is induced by phenobarbitone and this species is susceptible to trypsin. 2. After trypsin treatment of smooth membrane, some NADPH-cytochrome P-450 (cytochrome c) reductase activity remains and this remaining activity is enhanced by treatment with 0.05% deoxycholate, which renders the membranes permeable to macromolecules. In non-trypsin-treated control membranes the reductase activity is increased to a similar extent. These observations suggest an asymmetric distribution of NADPH-cytochrome P-450 (cytochrome c) reductase in the membrane. 3. As compared with dithionite, NADPH reduces only 44% of the cytochrome P-450 present in intact membranes. After tryptic digestion, none of the remaining cytochrome P-450 is reducible by NADPH. 4. In the presence of both a superoxide-generating system (xanthine plus xanthine oxidase) and NADPH, all the cytochrome P-450 in intact membrane (as judged by dithionite reducibility) is reduced. The cytochrome P-450 remaining after trypsin treatment of smooth vesicles cannot be reduced by this method. 5. The superoxide-dependent reduction of cytochrome P-450 is prevented by treatment of the membranes with mersalyl, which inhibits NADPH-cytochrome P-450 (cytochrome c) reductase. Thus the effect of superoxide may involve NADPH-cytochrome P-450 reductase and cytosolically orientated membrane factor(s).


Computed tomography of the knee.
Although magnetic resonance imaging (MRI) has become the imaging technique of choice in Canada and elsewhere in the evaluation of knee disorders, magnetic resonance units are frequently unavailable, or access to them is greatly restricted. The authors have found that computed tomography (CT) can sometimes be successfully substituted for MRI as a noninvasive method for imaging a wide variety of knee disorders. This article reviews the CT appearance of normal knee anatomy and illustrates a wide variety of pathological conditions that can be successfully evaluated.


Hypoxia-inducible-factors differentially contribute to clinical disease and the control of viral replication during RSV infection.
Hypoxia-inducible-factors (HIF) are transcription factors that regulate cellular adaptation to hypoxic conditions, enabling cells to survive in low-oxygen environments. Viruses have evolved to stabilize this pathway to promote successful viral infection, therefore modulation of HIFs could represent a novel antiviral strategy. In previous in vitro studies, we demonstrate respiratory syncytial virus (RSV), a leading cause of respiratory illness, to stabilize HIFs under normoxic conditions, with inhibition of HIF-1α resulting in reduced viral replication. Despite several HIF modulating compounds being tested/approved for use in other non-infectious models, little is known about their efficacy against respiratory viruses using relevant animal models. This study aimed to characterize the disease modulating properties and antiviral potential of anti-HIF-1α (PX478) and anti-HIF-2α (PT2385) in RSV-infected BALB/c mice. We found inhibition of HIF-1α to worsen clinical disease parameters, while simultaneously improving airway function. Additionally, anti-HIF-1α results in significantly reduced viral titer at early and peak time points of RSV replication, followed by a loss in viral clearance when given every day, but not every-other-day. In contrast, inhibition of HIF-2α was associated with improved clinical parameters, with no changes in airway function, and amelioration of interstitial pneumonia. Furthermore, anti-HIF-2α reduced early and peak lung viral replication, with no impaired viral clearance. Analysis of lung cells found significant modification in the T cell compartment that correlated with changes in lung pathology and viral titers in response to each HIF inhibitor administration. These data underscore the complex role of HIFs in RSV infection and highlight the need for careful therapeutic consideration.


[Anti-inflammatory effect of mineral water inhalation: validity of identification of biochemical markers in nasal secretion].
The study was made of therapeutic activity of silicon mineral water Tersinka inhalations on clinical symptoms, external respiration function and activity of inflammation in patients with chronic bronchitis and bronchial asthma as shown by biochemical markers of inflammation in the blood serum and nasal secretion. A correlation was found between treatment efficacy and concentration of sialic acids in the nasal secretion in pulmonological patients.


Use of horseradish peroxidase-labeled antibody for light and electron microscope localization of reovirus antigen.
Horseradish peroxidase-labeled antibody was used for light and electron microscopic localization of reovirus antigen in tissue culture. Reaction product in infected cells was easily detected in the cytoplasm, and the procedure was as sensitive as the fluorescent-antibody technique. At the electron microscopic level, infected and enzyme-labeled antibody-treated cells showed accumulations of reaction product at the sites of viral replication and around the viral particles. Reaction product was not detected in unstained infected cells, in stained uninfected cells, or in cells infected with an unrelated virus.


The Patient Experience: An Analysis of Orthopedic Surgeon Quality on Physician-Rating Sites.
With the advent of the Consensus Core of Orthopedic Measures, arthroplasty surgeons are increasingly subjected to public performance reviews on physician-rating sites. Therefore, we evaluated (1) web site details of physician-rating sites, (2) differences between sites and the Consensus Core, (3) published patient experiences, (4) search rank among sites, and (5) differences between academic vs nonacademic and arthroplasty vs nonarthroplasty surgeons. The 5 busiest physician-rating sites were analyzed. To compare physician-rating sites to the Consensus Core, 3 reviewers analyzed the web site details. To evaluate patient ratings and reviews, orthopedists from the top 5 academic and nonacademic hospitals (2016 US News & World Report) were analyzed. Institution-produced rating sites were also analyzed. Findings were stratified between academic vs nonacademic and arthroplasty vs nonarthroplasty surgeons. Five hundred and six staff surgeons across 10 academic and nonacademic affiliated hospitals yielded 27,792 patient-generated ratings and reviews for 1404 accounts. Features on all sites were practice location, languages spoken, and patient experience. Two sites autogenerated profiles of surgeons without consent. No physician-rating site contained all Consensus Core domains. The composite orthopedic surgeon rating was 4.1 of 5. No significant differences were found between academic and nonacademic affiliated surgeons. Arthroplasty surgeons had a greater number of reviews and ratings on 2 sites. Reliability of physician-rating sites is questionable, as none contained all Consensus Core domains. Autogeneration of surgeon profiles is occurring, and no differences between academic vs nonacademic or arthroplasty vs nonarthroplasty surgeons were found. Institution-produced sites may serve to better promote and market surgeons.


Using GIS to evaluate a fire safety program in North Carolina.
Evaluating program impact is a critical aspect of public health. Utilizing Geographic Information Systems (GIS) is a novel way to evaluate programs which try to reduce residential fire injuries and deaths. The purpose of this study is to demonstrate the application of GIS within the evaluation of a smoke alarm installation program in North Carolina. This approach incorporates national fire incident data which, when linked with program data, provides a clear depiction of the 10 years impact of the Get Alarmed, NC! program and estimates the number of potential lives saved. We overlapped Get Alarmed, NC! program installation data with national information on fires using GIS to identify homes that experienced a fire after an alarm was installed and calculated potential lives saved based on program documentation and average housing occupancy. We found that using GIS was an efficient and quick way to match addresses from two distinct sources. From this approach we estimated that between 221 and 384 residents were potentially saved due to alarms installed in their homes by Get Alarmed, NC!. Compared with other program evaluations that require intensive and costly participant telephone surveys and/or in-person interviews, the GIS approach is inexpensive, quick, and can easily analyze large disparate datasets. In addition, it can be used to help target the areas most at risk from the onset. These benefits suggest that by incorporating previously unutilized data, the GIS approach has the potential for broader applications within public health program evaluation.


Coronary ostial stenosis complicating coronary arteriography.
Coronary ostial stenosis is a cause of angina pectoris and sudden death. It is due mainly to atherosclerosis, syphilis, and iatrogenic disease. The last is of growing importance because of the frequency of coronary arteriography and surgical procedures on the aortic valve. Since both may cause stenosis of the coronary ostia, these procedures raise the topic from an obscure morphologic entity to an important consideration in the treatment of cardiac disease. We describe a case of ostial stenosis that was complicated by coronary arteriography.


IdentiCS--identification of coding sequence and in silico reconstruction of the metabolic network directly from unannotated low-coverage bacterial genome sequence.
A necessary step for a genome level analysis of the cellular metabolism is the in silico reconstruction of the metabolic network from genome sequences. The available methods are mainly based on the annotation of genome sequences including two successive steps, the prediction of coding sequences (CDS) and their function assignment. The annotation process takes time. The available methods often encounter difficulties when dealing with unfinished error-containing genomic sequence. In this work a fast method is proposed to use unannotated genome sequence for predicting CDSs and for an in silico reconstruction of metabolic networks. Instead of using predicted genes or CDSs to query public databases, entries from public DNA or protein databases are used as queries to search a local database of the unannotated genome sequence to predict CDSs. Functions are assigned to the predicted CDSs simultaneously. The well-annotated genome of Salmonella typhimurium LT2 is used as an example to demonstrate the applicability of the method. 97.7% of the CDSs in the original annotation are correctly identified. The use of SWISS-PROT-TrEMBL databases resulted in an identification of 98.9% of CDSs that have EC-numbers in the published annotation. Furthermore, two versions of sequences of the bacterium Klebsiella pneumoniae with different genome coverage (3.9 and 7.9 fold, respectively) are examined. The results suggest that a 3.9-fold coverage of the bacterial genome could be sufficiently used for the in silico reconstruction of the metabolic network. Compared to other gene finding methods such as CRITICA our method is more suitable for exploiting sequences of low genome coverage. Based on the new method, a program called IdentiCS (Identification of Coding Sequences from Unfinished Genome Sequences) is delivered that combines the identification of CDSs with the reconstruction, comparison and visualization of metabolic networks (free to download at http://genome.gbf.de/bioinformatics/index.html). The reversed querying process and the program IdentiCS allow a fast and adequate prediction protein coding sequences and reconstruction of the potential metabolic network from low coverage genome sequences of bacteria. The new method can accelerate the use of genomic data for studying cellular metabolism.


Delayed auditory pathway maturation in the differential diagnosis of hypoacusis in young children.
Auditory deficiencies have profound implications for the development of communication abilities, social exchange, and cognitive skills. Although advances in technological instrumentation have decreased the age at which it is possible to detect auditory deficits, an absence of electrophysiological or behavioral reaction to sound does not always reflect hypoacusis. The objective of this study was to evaluate the evolution of audition in a group of children referred for therapy to the CINDA Foundation (Center for Research and Information in Auditory Deficiencies) in Bogota, Colombia, with an initial diagnosis of severe to profound sensorineural hearing loss. We evaluated 25 patients younger than 2.5 years of age with a protocol which included auditory evoked potentials, behavioral responses (tonal audiometry-responses to voice and startle) and language observation. After evaluation, the children began a program of auditory stimulation with audiological follow-up for a period of 6 months to 1 year. The audiological follow-up of the children after auditory stimulation showed significant improvements in both electrophysiological and behavioral indices of their hearing levels (p<.001, one-tailed Wilcoxon exact test). We found a high percentage of children in early stages of auditory maturation with a false diagnosis of hypoacusis (32%). The absence of response may result from compromised auditory reception or processing. Thus, delayed maturation of the auditory pathway should be considered in the differential diagnosis of young children with sensorineural hearing loss.


BAC-End Sequence-Based SNP Mining in Allotetraploid Cotton (Gossypium) Utilizing Resequencing Data, Phylogenetic Inferences, and Perspectives for Genetic Mapping.
A bacterial artificial chromosome library and BAC-end sequences for cultivated cotton (Gossypium hirsutum L.) have recently been developed. This report presents genome-wide single nucleotide polymorphism (SNP) mining utilizing resequencing data with BAC-end sequences as a reference by alignment of 12 G. hirsutum L. lines, one G. barbadense L. line, and one G. longicalyx Hutch and Lee line. A total of 132,262 intraspecific SNPs have been developed for G. hirsutum, whereas 223,138 and 470,631 interspecific SNPs have been developed for G. barbadense and G. longicalyx, respectively. Using a set of interspecific SNPs, 11 randomly selected and 77 SNPs that are putatively associated with the homeologous chromosome pair 12 and 26, we mapped 77 SNPs into two linkage groups representing these chromosomes, spanning a total of 236.2 cM in an interspecific F2 population (G. barbadense 3-79 × G. hirsutum TM-1). The mapping results validated the approach for reliably producing large numbers of both intraspecific and interspecific SNPs aligned to BAC-ends. This will allow for future construction of high-density integrated physical and genetic maps for cotton and other complex polyploid genomes. The methods developed will allow for future Gossypium resequencing data to be automatically genotyped for identified SNPs along the BAC-end sequence reference for anchoring sequence assemblies and comparative studies.


Comparative sensitivity of harbour and grey seals to several environmental contaminants using in vitro exposure.
In this study, we investigated the effects of cadmium chloride (CdCl(2)), mercury chloride (HgCl(2)), methylmercury chloride (CH(3)HgCl), and PCBs on lymphocyte proliferation in phocids. PBMCs isolated from harbour and grey seals were exposed in vitro to varying concentrations of contaminants. A reduction of viability occurred when cells were exposed to 10(-4)M HgCl(2) or CH(3)HgCl or to 50ppm of Aroclor 1254. In both grey and harbour seals, T-lymphocyte proliferation was suppressed when their cells were incubated with 5 x 10(-5)M CdCl(2) or 10(-4)M HgCl(2). An inhibition of proliferation occurred with CH(3)HgCl from 10(-6)M in grey seals and from 10(-5)M in harbour seals. In grey seals, Aroclor 1254 reduced lymphocyte proliferation at 15ppm. In both harbour and grey seals, CH(3)HgCl was ten times more immunotoxic that HgCl(2). From IC(50), chemicals were ranked in terms of toxicity as followed: CH(3)HgCl>CdCl(2)>HgCl(2)>Aroclor 1254.


Peripheral and central nervous system correlates in fibromyalgia.
Fibromyalgia (FM) is a syndrome characterized by altered pain processing at central and peripheral level, whose pathophysiologic mechanisms remain obscure. We aimed at exploring the structural changes of peripheral nociceptor measured by skin biopsy, the functional changes of central nociceptive pathway assessed by laser-evoked potentials (LEP), and their correlation with clinical features and comorbidities. In all, 81 patients diagnosed with FM underwent skin biopsies with quantification of intraepidermal nerve fibre density (IENFD) at the thigh and distal leg, and LEP recording by stimulating hand, thigh and foot. Nerve conduction study (NCS), clinical features, comorbidity with migraine and mood disorders, and previous, non-active immune-mediated disorders were recorded. Intraepidermal nerve fibre density was reduced in 85% of patients at the thigh and in 12.3% of patients at the distal leg, whereas it was normal in 14.8% of patients. N2P2 habituation index from laser stimulation at the thigh was altered in 97.5% of patients and correlated with reduced IENFD at the thigh. LEP latencies and amplitudes did not differ among groups. No association was found between IENFD, LEP, clinical features and comorbidities. Fibromyalgia patients most commonly showed a mild loss of peripheral nociceptors at the thigh rather than distal small fibre neuropathy. This finding was associated with an altered habituation index and strengthened the hypothesis that central sensitization plays a key role in the pathogenesis of the disease. Central impairment of pain processing likely underlies FM, which in most patients is associated with mild proximal small fibre pathology.


Multiple vibration displacements at multiple vibration frequencies stress impact on human femur computational analysis.
Whole-body vibration training using single-frequency methods has been reported to improve bone mineral density. However, the intensities can exceed safe levels and have drawn unfavorable comments from subjects. In a previous article, whole-body vibration training using multiple vibration displacements at multiple vibration frequencies (MVDMVF) was reported. This article presents the computational simulation evaluation of stress dispersion on a femur with and without the MVDMVF input. A model of bone femur was developed from a computed tomography image of the lower limb with Mimics software from Materialise (Plymouth, Michigan). We analyzed the mesh model in COMSOL Multiphysics (COMSOL, Inc; Burlington, Massachusetts) with and without MVDMVF input, with constraints and load applied to the femur model. We compared the results with published joint stresses during walking, jogging, and stair-climbing and descending and with standard vibration exposure limits. Results showed stress levels on the femur are significantly higher with MVDMVF input than without. The stress levels were within the published levels during walking and stair-climbing and descending but below the stress levels during jogging. Our computational results demonstrate that MVDMVF generates stress level equivalent to the level during walking and stair-climbing. This evidence suggests that MVDMVF is safe for prolonged use in subjects with osteoporosis who ambulate independently.


Stabilization of the 3' one-third of Escherichia coli ribosomal protein S20 mRNA in mutants lacking polynucleotide phosphorylase.
Mutations which largely inactivate polynucleotide phosphorylase and which render RNase II thermolabile exert two effects on the metabolism of the two nested mRNAs which encode ribosomal protein S20. (i) The lifetime of both mRNA species is extended 2.5-fold at 38 degrees C in a strain harboring both mutations. (ii) A relatively stable truncated fragment of these mRNAs accumulates to significant levels in strains lacking polynucleotide phosphorylase. The truncated RNA (Po RNA) is 147 to 148 residues long and is coterminal with the 3' ends of intact S20 mRNAs. Its 5' end appears to be generated by endonucleolytic cleavage to the 5' side of a G residue in the sequence AACCGAUC. The data are consistent with the hypothesis that S20 mRNAs can be degraded by alternative pathways. The normal pathway depends on functional polynucleotide phosphorylase and is concerted, since S20 mRNAs disappear without accumulation of detectable intermediates in the decay process. The slower alternative pathway is followed when polynucleotide phosphorylase is inactivated by mutation. This pathway is distinguished by segmental rather than concerted degradation of S20 mRNAs and involves at least one endonucleolytic cleavage. The 5' two-thirds of S20 mRNAs decays significantly more quickly than the 3' third in this latter mode of mRNA turnover.


Protective effects of ginger-turmeric rhizomes mixture on joint inflammation, atherogenesis, kidney dysfunction and other complications in a rat model of human rheumatoid arthritis.
Besides joint destruction, extra-articular complications (outside the locomotor system) are frequent in rheumatoid arthritis (RA) patients, especially cardiovascular, hematological and metabolic disorders. Here, we evaluated and compared the protective activity of two different doses of mixture of ginger and turmeric rhizomes powder (1 : 1) suspended in distilled water (GTaq) in alleviating both articular and extra-articular manifestations in rat adjuvant-induced arthritis (AIA). Arthritis was induced by a single intra-dermal injection of 0.1 mL of Complete Freund's adjuvant (containing heat-killed Mycobacterium tuberculosis) into the palmar surface of the left hind paw after the rats were subjected to light diethyl ether anesthesia. Arthritic rats received orally and daily (for 28 consecutive days) distilled water as vehicle, indomethacin (1.0 mg/kg body weight), or GTaq (200 or 400 mg/kg body weight) from the day of arthritis induction. The present study showed that GTaq (especially the high dose) was more effective (4.2-38.4% higher, P < 0.05-0.001) than indomethacin (a non-steroidal/anti-inflammatory drug) in alleviating the loss in body weight gain, the histopathological changes observed in ankle joints, blood leukocytosis and thrombocytosis, iron deficiency anemia, serum hypoalbuminemia and globulinemia, the impairment of kidney functions, and the risks for cardiovascular disease in arthritic rats. These protective effects of GTaq were mediated through increasing the food intake and decreasing the systemic inflammation that occur at the appearance of polyarthritis, oxidative stress and dyslipidemia. Ginger-turmeric rhizomes mixture may be effective against RA severity and complications as shown in an AIA rat model.