Electroless deposition of Ni-P/Au coating on Cu substrate with improved corrosion resistance from Au(iii)-DMH based cyanide-free plating bath using hypophosphite as a reducing agent.
The green cyanide-free gold deposition is an important development direction in electroless gold plating. However, the commonly Au(i) based cyanide-free gold plating bath always suffers from severe corrosion to the Ni-P layer and unsatisfactory service life of the bath. In this work, a green and environmentally friendly cyanide-free gold plating bath was developed with hypophosphite added as a reducing agent into the Au(iii)-DMH (5,5-dimethylhydantoin) based plating bath to retard the corrosion of the Ni-P layer. SEM micrographs combined with XRD and XPS analysis indicated that the electroless deposited gold was pure and compact. And XRD also revealed that the oriented deposition of gold was growing preferentially on Au (111). Corrosion tests, including salt spray tests, potentiodynamic polarization tests, and electrochemical impedance spectroscopy tests, indicated that the obtained Cu/Ni-P/Au coatings had significantly improved corrosion resistance performance with the loading of hypophosphite as the reducing agent. The baths remained transparent and no turbidity or precipitates were detected for 210 days, reflecting good stability. The detection for the Ni2+ concentration in the bath showed that adding hypophosphite could retard the replacement reaction between Au3+ and the Ni-P layer in part and this is important for decreasing the severe corrosion of the Ni-P layer. Moreover, Au was inactive for catalyzing the oxidation of hypophosphite during the deposition process which was confirmed furthermore via quantum chemical calculations. Therefore, our developed green cyanide-free electroless gold deposition process can beneficially provide research value and application prospects in microelectronic industry.


Computerized tomography in obstructive sleep apnea. Correlation of airway size with physiology during sleep and wakefulness.
Pathophysiologic changes during sleep in patients with obstructive apnea are often associated with alterations in upper airway function during awake periods. To determine whether these functional changes are related to abnormal airway structure, we performed computerized tomography (CT) in 20 awake patients with obstructive apnea and in 10 control subjects. The CT scan measurements of cross-sectional areas of the nasopharynx, oropharynx, and hypopharynx in apneic patients were significantly reduced (p less than 0.05) compared with those in the control subjects. Sites of obstruction varied, and apparent airway occlusion occurred in 6 patients. Reduced pharyngeal size correlated with increased sleep-disordered breathing rates (p less than 0.05), more severe nocturnal oxygen desaturation (p less than 0.05), and the presence of a saw-tooth pattern of flow-volume curves obtained during awake periods (p less than 0.05). Structural upper airway narrowing is detectable with CT in awake patients with obstructive sleep apnea.


Microbial degradation of naphthenic acids using constructed wetland treatment systems: metabolic and genomic insights for improved bioremediation of process-affected water.
Naphthenic acids (NAs) are a complex mixture of organic compounds released during bitumen extraction from mined oil sands that are important contaminants of oil sands process-affected water (OSPW). NAs can be toxic to aquatic organisms and, therefore, are a main target compound for OSPW. The ability of microorganisms to degrade NAs can be exploited for bioremediation of OSPW using constructed wetland treatment systems (CWTS), which represent a possible low energy and low-cost option for scalable in situ NA removal. Recent advances in genomics and analytical chemistry have provided insights into a better understanding of the metabolic pathways and genes involved in NA degradation. Here, we discuss the ecology of microbial NA degradation with a focus on CWTS and summarize the current knowledge related to the metabolic pathways and genes used by microorganisms to degrade NAs. Evidence to date suggests that NAs are mostly degraded aerobically through ring cleavage via the beta-oxidation pathway, which can be combined with other steps such as aromatization, alpha-oxidation, omega-oxidation, or activation as coenzyme A (CoA) thioesters. Anaerobic NA degradation has also been reported via the production of benzoyl-CoA as an intermediate and/or through the involvement of methanogens or nitrate, sulfate, and iron reducers. Furthermore, we discuss how genomic, statistical, and modeling tools can assist in the development of improved bioremediation practices.


Topical application of L-menthol induces heat analgesia, mechanical allodynia, and a biphasic effect on cold sensitivity in rats.
Menthol is used in analgesic balms and also in foods and oral hygiene products for its fresh cooling sensation. Menthol enhances cooling by interacting with the cold-sensitive thermoTRP channel TRPM8, but its effect on pain is less well understood. We presently used behavioral methods to investigate effects of topical menthol on thermal (hot and cold) pain and innocuous cold and mechanical sensitivity in rats. Menthol dose-dependently increased the latency for noxious heat-evoked withdrawal of the treated hindpaw with a weak mirror-image effect, indicating antinociception. Menthol at the highest concentration (40%) reduced mechanical withdrawal thresholds, with no effect at lower concentrations. Menthol had a biphasic effect on cold avoidance. At high concentrations (10% and 40%) menthol reduced avoidance of colder temperatures (15 degrees C and 20 degrees C) compared to 30 degrees C, while at lower concentrations (0.01-1%) menthol enhanced cold avoidance. In a -5 degrees C cold plate test, 40% menthol significantly increased the nocifensive response latency (cold hypoalgesia) while lower concentrations were not different from vehicle controls. These results are generally consistent with neurophysiological and human psychophysical data and support TRPM8 as a potential peripheral target of pain modulation.


Structural pursuit over multiple undirected graphs.
Gaussian graphical models are useful to analyze and visualize conditional dependence relationships between interacting units. Motivated from network analysis under di erent experimental conditions, such as gene networks for disparate cancer subtypes, we model structural changes over multiple networks with possible heterogeneities. In particular, we estimate multiple precision matrices describing dependencies among interacting units through maximum penalized likelihood. Of particular interest are homogeneous groups of similar entries across and zero-entries of these matrices, referred to as clustering and sparseness structures, respectively. A non-convex method is proposed to seek a sparse representation for each matrix and identify clusters of the entries across the matrices. Computationally, we develop an e cient method on the basis of di erence convex programming, the augmented Lagrangian method and the block-wise coordinate descent method, which is scalable to hundreds of graphs of thousands nodes through a simple necessary and sufficient partition rule, which divides nodes into smaller disjoint subproblems excluding zero-coe cients nodes for arbitrary graphs with convex relaxation. Theoretically, a finite-sample error bound is derived for the proposed method to reconstruct the clustering and sparseness structures. This leads to consistent reconstruction of these two structures simultaneously, permitting the number of unknown parameters to be exponential in the sample size, and yielding the optimal performance of the oracle estimator as if the true structures were given a priori. Simulation studies suggest that the method enjoys the benefit of pursuing these two disparate kinds of structures, and compares favorably against its convex counterpart in the accuracy of structure pursuit and parameter estimation.


Simplified Attachable EEG Revealed Child Development Dependent Neurofeedback Brain Acute Activities in Comparison with Visual Numerical Discrimination Task and Resting.
The development of an easy-to-attach electroencephalograph (EEG) would enable its frequent use for the assessment of neurodevelopment and clinical monitoring. In this study, we designed a two-channel EEG headband measurement device that could be used safely and was easily attachable and removable without the need for restraint or electrode paste or gel. Next, we explored the use of this device for neurofeedback applications relevant to education or neurocognitive development. We developed a prototype visual neurofeedback game in which the size of a familiar local mascot changes in the PC display depending on the user's brain wave activity. We tested this application at a local children's play event. Children at the event were invited to experience the game and, upon agreement, were provided with an explanation of the game and support in attaching the EEG device. The game began with a consecutive number visual discrimination task which was followed by an open-eye resting condition and then a neurofeedback task. Preliminary linear regression analyses by the least-squares method of the acquired EEG and age data in 30 participants from 5 to 20 years old suggested an age-dependent left brain lateralization of beta waves at the neurofeedback stage (p = 0.052) and of alpha waves at the open-eye resting stage (p = 0.044) with potential involvement of other wave bands. These results require further validation.


Acidification remediation alternatives: exploring the temporal dimension with cost benefit analysis.
Acidification of soils and surface waters caused by acid deposition is still a major problem in southern Scandinavia, despite clear signs of recovery. Besides emission control, liming of lakes, streams, and wetlands is currently used to ameliorate acidification in Sweden. An alternative strategy is forest soil liming to restore the acidified upland soils from which much acidified runoff originates. This cost-benefit analysis compared these liming strategies with a special emphasis on the time perspective for expected benefits. Benefits transfer was used to estimate use values for sport ffishing and nonuse values in terms of existence values. The results show that large-scale forest soil liming is not socioeconomically profitable, while lake liming is, if it is done efficiently-in other words, if only acidified surface waters are treated. The beguiling logic of "solving" an environmental problem at its source (soils), rather than continuing to treat the symptoms (surface waters), is thus misleading.


[Features of the epidemiologic characteristics of ischemic heart disease among scientific research workers].
The peculiarities of prevalence of coronary heart disease (CHD), its main risk factors and their interrelationships were studied on the basis of the results of epidemiological surveys of an organized population of men aged 20 to 59 engaged in research, and an unorganized male population in one of Moscow districts. A similar CHD frequency was revealed in both populations (10.5 and 9.5%), however the prevalence of possible CHD was significantly higher among the research workers. In this group more unfavorable epidemiological conditions were noted with relation to such CHD risk factors as hypercholesterolemia, disturbed glucose tolerance, a low physical activity, with practically the same prevalence of excess body mass and lower frequency of arterial hypertension and smoking. The results obtained formed the basis for a design of measures on primary and secondary CHD prevention among research workers.


Circulating RNA transcripts identify therapeutic response in cystic fibrosis lung disease.
Circulating leukocyte RNA transcripts are systemic markers of inflammation, which have not been studied in cystic fibrosis (CF) lung disease. Although the standard assessment of pulmonary treatment response is FEV(1), a measure of airflow limitation, the lack of systemic markers to reflect changes in lung inflammation critically limits the testing of proposed therapeutics. We sought to prospectively identify and validate peripheral blood leukocyte genes that could mark resolution of pulmonary infection and inflammation using a model by which RNA transcripts could increase the predictive value of spirometry. Peripheral blood mononuclear cells were isolated from 10 patients with CF and acute pulmonary exacerbations before and after therapy. RNA expression profiling revealed that 10 genes significantly changed with treatment when compared with matched non-CF and control subjects with stable CF to establish baseline transcript abundance. Peripheral blood mononuclear cell RNA transcripts were prospectively validated, using real-time polymerase chain reaction amplification, in an independent cohort of acutely ill patients with CF (n = 14). Patients who responded to therapy were analyzed using general estimating equations and multiple logistic regression, such that changes in FEV(1)% predicted were regressed with transcript changes. Three genes, CD64, ADAM9, and CD36, were significant and independent predictors of a therapeutic response beyond that of FEV(1) alone (P < 0.05). In both cohorts, receiver operating characteristic analysis revealed greater accuracy when genes were combined with FEV(1). Circulating mononuclear cell transcripts characterize a response to the treatment of pulmonary exacerbations. Even in small patient cohorts, changes in gene expression in conjunction with FEV(1) may enhance current outcomes measures for treatment response.


What Is the Outcome of Allograft and Intramedullary Free Fibula (Capanna Technique) in Pediatric and Adolescent Patients With Bone Tumors?
After bone tumor resection, reconstruction for limb salvage surgery can be challenging because of the resultant large segmental bony defects. Structural allografts have been used to fill these voids; however, this technique is associated with high complication rates. To circumvent the complications associated with this procedure, massive bony allografts have been supplemented with an intramedullary vascularized free fibula. However, few studies have examined the outcomes using this technique in the pediatric and adolescent populations. The purpose of this study was to examine the revision-free survival using he Capanna technique for limb salvage for pediatric lower limb salvage. We attempted to answer the following questions: (1) What was the overall limb salvage rate along with incidence of reoperation and complications? (2) How did pediatric and adolescent patients functionally perform after this technique? (3) What was the incidence of late complications including infection and fracture? (4) What was the incidence of limb length discrepancy? Eighteen pediatric patients who underwent lower extremity limb salvage with the use of cadaveric allograft and intramedullary free fibular transfer (Capanna technique) were identified. There were nine boys males and nine girls with a mean age of 11 years (range, 5-18 years) and mean followup of 8 years (range, 2-15 years), respectively. All patients had at least 2 years followup. Three patients have not been seen in followup during the past 5-years; however, all had made it to their 5-year clinical followup. The patients' medical records were reviewed for clinical and functional outcomes as well as postoperative complications. Time to union was recorded through an evaluation of radiographs. Mankin functional outcome and Musculoskeletal Tumor Society (MSTS) rating scale were recorded for each patient. The overall limb salvage rate was 94%. Fourteen patients underwent an additional surgical procedure. Six patients underwent additional procedure(s) to treat a symptomatic nonunion. Seventeen of the patients had a good or excellent Mankin score with a mean MSTS rating of 93% at last followup. Six of the patients underwent a limb length modification procedure. Use of large allografts in conjunction with intramedullary vascularized free fibulas appears to be a reliable method for the reconstruction of large bony tumors of the lower extremity in this population, although we did not directly compare this with allografts alone in our study. The use of locked plates may improve union times. The proportion of patients achieving limb preservation was high and complication rates are acceptable. Level IV, therapeutic study.


Restorative proctocolectomy for ulcerative colitis complicated by colorectal cancer.
Restorative proctocolectomy with ileal pouch-anal anastomosis is widely accepted as the procedure of choice for patients requiring surgery for chronic ulcerative colitis. The role of restorative proctocolectomy in the setting of chronic ulcerative colitis complicated by colorectal carcinoma is not clear. This study was undertaken to explore the clinical outcomes of chronic ulcerative colitis patients with coexisting colorectal carcinoma who underwent restorative proctocolectomy. A total of 756 patients with chronic ulcerative colitis were followed prospectively after restorative proctocolectomy. Forty-five (5.9 percent) were found to have invasive carcinoma of the colon (n = 31) or rectum (n = 14). These patients were followed with special attention to cancer stage, adjuvant therapy, oncologic outcome, and functional results after restorative proctocolectomy. Twenty-one patients (45.6 percent) had staged surgery (colon, 14; rectum, 7). Twenty-seven patients received adjuvant chemotherapy (colon, 22; rectum, 5). Fourteen patients (51.8 percent) who received chemotherapy were not diverted during this treatment. Two node-positive rectal cancer patients had pelvic radiotherapy: one before restorative proctocolectomy and one after restorative proctocolectomy. Mean time to restoration of intestinal continuity among staged patients did not differ between cancer and noncancer patients. Six patients died of metastatic disease (colon, 3; rectum, 3). Five deaths occurred among patients with Stage III disease (colon, 3/13, 23.1 percent; rectum, 2/3, 66.7 percent). One patient with Stage I cancer at the time of restorative proctocolectomy died. Thirty-nine patients are alive without evidence of disease at a mean interval from surgery of 76.5 months. Thirty-six patients have functioning pelvic pouches. Bowel frequency, continence, and complication rates are similar among restorative proctocolectomy patients with and without cancer. Restorative proctocolectomy as a single or staged procedure is a viable therapeutic option for selected chronic ulcerative colitis patients with associated colorectal cancers. Prognosis seems to be related to cancer stage. Adjuvant chemotherapy can safely be given to nondiverted patients. Appropriate use of preoperative and postoperative radiotherapy for rectal cancer patients who are otherwise candidates for restorative proctocolectomy is unknown. Long-term functional results for cancer patients are similar to those seen in chronic ulcerative colitis patients without cancer.


Stability of self-prepared fortified antibiotic eyedrops.
Self-prepared fortified antibiotic eye drops are essential for the severe ocular infection. The relation of potency decay and storage conditions including temperature, concentration and duration were studied by the changes of MIC. 1% and 10% amikin, 10% and 50% pipril, and 5% and 25% vancomycin were diluted from the prarenteral antibiotics with the 5% glucose and storage at 4, -18 and -80 degrees C for 3, 7, 14 and 28 days. MICs of amikin against Pseudomonas aeruginosa, pipril and vancomycin against Staphylococcus aureus were determined by the agar diffusion method. Fluctuations of the MIC were noted during the observation period. Most of the significant changes of MIC were found during the first 7 days. When the potencies between time of zero and 28 days are compared, only 10% pipril and 25% vancomycin stored at -80 degrees C had significant change. Our conclusion is that all of the fortified antibiotic eye drops in this study can be stored in the house refrigerator or freezer for up to 28 days. High concentration may show a negative result of the preservation. Temperature does not influence the preservative effect within four weeks' observation.


Neuroprotective Effect of Perilla Extracts on PC12 Cells.
Perilla frutescens (L) Britton contains some active principles which had neuroprotective actions. The present study was aimed to investigate the effect of Perilla extracts on neuroprotection, antioxidation and neurite outgrowth in PC12 cells. The neuroprotective effect of freeze-dried ethanolic extract from Perilla leaves and cold-pressed seed oil were tested on PC12 induced with beta-amyloid protein. The inhibition of tau-protein hyperphosphorylation and the antioxidant enzyme activity were analyzed. The neurite outgrowth bearing cells were investigated and MEK-1 protein production was analyzed by enzyme immunometric assay. In PC12 culture induced toxicity by beta-amyloid protein: (1) the decrease in cell viability was attenuated in cells pretreated with leaf extract 200 mg/ml and oil 50 mg/ml; (2) SOD activity seemed to decrease when pretreated cells with the extracts; (3) tau phosphorylation was decreased by pretreated cells with50 mg/ml of oil. Moreover, given Perilla leaf extract or seed oil to PC12 culture, the amount of neurite outgrowth bearing cells increased harmoniously with MEK-1 protein expression. Perilla leaf extract and seed oil reversed the effect of beta-amyloid induced toxicity by decreasing oxidative stress and inhibition of tau-protein hyperphosphorylation. The enhancement of neurite outgrowth by Perilla extracts was also revealed.


Electrosensory ampullary organs are derived from lateral line placodes in cartilaginous fishes.
Ampullary organ electroreceptors excited by weak cathodal electric fields are used for hunting by both cartilaginous and non-teleost bony fishes. Despite similarities of neurophysiology and innervation, their embryonic origins remain controversial: bony fish ampullary organs are derived from lateral line placodes, whereas a neural crest origin has been proposed for cartilaginous fish electroreceptors. This calls into question the homology of electroreceptors and ampullary organs in the two lineages of jawed vertebrates. Here, we test the hypothesis that lateral line placodes form electroreceptors in cartilaginous fishes by undertaking the first long-term in vivo fate-mapping study in any cartilaginous fish. Using DiI tracing for up to 70 days in the little skate, Leucoraja erinacea, we show that lateral line placodes form both ampullary electroreceptors and mechanosensory neuromasts. These data confirm the homology of electroreceptors and ampullary organs in cartilaginous and non-teleost bony fishes, and indicate that jawed vertebrates primitively possessed a lateral line placode-derived system of electrosensory ampullary organs and mechanosensory neuromasts.


Gestational Cocaine Exposure Alters Postnatal Pituitary-Adrenal Axis Activity and Stress Endurance in Ratsa.
Female pregnant Sprague-Dawley rats were injected once daily with 40 mg/kg cocaine hydrochloride or 0.9% saline from gestational day 12 (GD 12) to GD 21. From postnatal day 21 (PND 21) to PND 60, both male and female offspring were examined for stress response. Treated male and female offspring demonstrated a diminished tolerance to stress as determined by a cold water stress test performed at PND 21, 30 and 40. Base hormonal levels of adrenocorticotropin hormone (ACTH) and corticosterone were not affected by prenatal cocaine exposure in male offspring at PND 30. However, immobilization for 1 hr caused a significant sustained elevation of corticosterone levels at both PND 30 and PND 60 in male treated offspring as compared to the control group. Plasma ACTH levels were also significantly sustained after 1 hr of immobilization at PND 60 for the cocaine-treated male offspring. These results indicate both a diminished capacity to respond to stress and an abnormal heightened reactivity of the pituitary-adrenal axis in offspring exposed to cocaine in utero.


Growth performance, bile acid profile, fecal microbiome and serum metabolomics of growing-finishing pigs fed diets with bile acids supplementation.
The present experiment was conducted to determine the effect of bile acids (BAs) supplementation on growth performance, BAs profile, fecal microbiome, and serum metabolomics in growing-finishing pigs. A total of 60 pigs [Duroc × (Landrace × Yorkshire)] with an average body weight of 27.0 ± 1.5 kg were selected and allotted into one of 2 groups (castrated male to female ratio = 1:1), with 10 replicates per treatment and 3 pigs per replicate. The 2 treatments were the control group (control) and a porcine bile extract-supplemented group dosed at 0.5 g/kg feed (BA). After a 16-wk treatment, growth performance, BAs profiles in serum and feces, and fecal microbial composition were determined. An untargeted metabolomics approach using gas chromatography with a time-of-flight mass spectrometer was conducted to identify the metabolic pathways and associated metabolites in the serum of pigs. We found that BAs supplementation had no effect on the growth performance of the growing-finishing pig. However, it tended to increase the gain-to-feed ratio for the whole period (P = 0.07). BAs supplementation resulted in elevated serum concentrations of secondary bile acids, including hyodeoxycholic acid (HDCA), glycoursodeoxycholic acid, and tauro-hyodeoxycholic acid, as well as fecal concentration of HDCA (P < 0.05). Fecal microbiota analysis revealed no differences in alpha and beta diversity indices or the relative abundance of operational taxonomic units (OTUs) at both phylum and genus levels between groups. Metabolic pathway analysis revealed that the differential metabolites between control and BA groups are mainly involved in purine metabolism, ether lipid metabolism, glycerophospholipid metabolism, and amino sugar and nucleotide sugar metabolism, as well as primary bile acid biosynthesis. Our findings indicate that BAs supplementation tended to improve the feed efficiency, and significantly altered the BA profile in the serum and feces of growing-finished pigs, regardless of any changes in the gut microbial composition. The altered metabolic pathways could potentially play a vital role in improving the feed efficiency of growing-finished pigs with BAs supplementation.


Clinical outcomes of implant-supported prosthetic rehabilitation of severely atrophic maxilla: A systematic review.
The purpose of this systematic review is to evaluate the clinical outcomes for the various methods of rehabilitation of a severely atrophic maxilla with the help of implant-supported fixed prosthesis. The relevant publications published between 2013 and 2022 and written only in English were identified using an electronic search. The primary research question for this study was developed based on the PICO framework, which stands for population, intervention, control, and outcomes which was "What are the clinical outcomes of implant-supported prosthetic rehabilitation in patients with severely atrophic maxilla?" The relevancy of the articles was confirmed by examining their titles, abstracts, and complete texts to determine whether they satisfied the requirements for inclusion. Utilizing specialized study design-related bias assessment forms, the risk of bias was evaluated. The database search resulted in 1568 results; however, 1529 of them were eliminated because of insufficient, duplicate, or missing data. Additionally, manual searching yielded 11 articles. After 50 full-text papers were assessed for eligibility, 17 articles were eliminated. Thus, 33 studies in total are included in the current systematic review. Risk of bias analysis and GRADE evidence analysis were performed. Data were found to be heterogeneous and thus meta-analysis could not be done and narrative synthesis is presented. The patient's condition and the clinician's expertise play a role in taking the decision on choice of technique for the fixed implant-supported rehabilitation of the severely atrophic maxilla. A high success and survival rate is produced by the majority of fixed implant-assisted prostheses despite the biologic and prosthetic problems. A single approach cannot be recommended as the gold standard. The choice is dependent on the patient's biological factors as well as the clinician's expertise. The included studies were assessed using GRADE criteria. The quality of evidence is low-medium. Therefore, to better comprehend the clinical effectiveness of the treatment alternatives, more well-designed randomized controlled trials with longer follow-up period are required.


Distribution and function of recombinant endothelial nitric oxide synthase in transplanted hearts.
Introducing recombinant genes into donor hearts may offer a therapeutic intervention that could potentially attenuate the complications of heart transplantation, including rejection, infection and accelerated atherosclerosis. In the cardiovascular system, reduced bioactivity of endothelial nitric oxide is a feature of atherosclerosis and vascular injury. Nitric oxide is an arterial vasodilator that also inhibits proliferation of vascular smooth muscle cells and platelet aggregation. Experiments were designed to determine the distribution of adenoviral-mediated transfer of recombinant endothelial nitric oxide synthase gene (eNOS) and the effect of recombinant gene expression on the function of transplanted hearts. Adenoviral vectors for (a) bovine eNOS (AdeNOS) or (b) beta-galactosidase (AdLacZ; control) were infused into two groups (n = 12, per group) of explanted rat hearts. The transduced hearts were then implanted heterotopically into the abdomen of syngeneic recipient rats. After four days, the hearts were excised and examined for distribution and function of the recombinant genes. Polymerase chain reaction (PCR) verified the presence of the recombinant eNOS gene in eNOS-transduced but not in beta-galactosidase-transduced hearts; reverse transcriptase-PCR identified mRNA for eNOS in AdeNOS-transduced hearts. NOS activity (conversion of tritiated L-arginine to citrulline) was greater in homogenates of AdeNOS-compared to AdLacZ-transduced hearts. Positive immunoreactivity for eNOS was present in cardiomyocytes predominantly in eNOS-transduced hearts. Myocardial contractility and coronary blood flow, as determined using a Langendorff preparation, were not different between hearts transduced with AdeNOS or AdLacZ. These results suggest that, up to four days post transplantation, adenoviral-mediated transfer of eNOS into transplanted hearts is possible. However, expression of the recombinant protein did not result in measurable changes in myocardial contractility or coronary perfusion.


Inhibition of allergic encephalomyelitis in rats by treatment with sulfated polysaccharides.
A number of sulfated polysaccharides were tested for their ability to inhibit passively induced experimental allergic encephalomyelitis (EAE) in rats. Heparin and fucoidan both completely inhibited passive EAE even when treatment was begun 3 days after transfer of cells. Pentosan sulfate was partially inhibitory whereas chondroitin-4-sulfate had no effect. Inhibition was not merely due to killing of the cells since active sensitization 14 days after cell transfer resulted in an early onset of disease indicating the persistence of transferred cells as memory cells. Although all the inhibitory polysaccharides are anticoagulants, it would appear that this function alone is not the reason for inhibition since a heparin preparation devoid of anticoagulant activity also partially inhibited EAE. Actively induced EAE was also significantly delayed by treatment with heparin. The results are discussed in terms of the polysaccharides inhibiting the enzymatic dependent movement of lymphocytes across central nervous system vascular endothelium.


The reliability of topographic measurements from navigated transcranial magnetic stimulation in healthy volunteers and tumor patients.
Navigated transcranial magnetic stimulation (nTMS) is increasingly being used for preoperative mapping of the motor cortex. Any new technology should undergo rigorous validation before being widely adopted in routine clinical practice. The aim of this experimental study was to assess the intraexaminer and interexaminer reliability of topographic mapping with nTMS. nTMS mapping of the motor cortex for the first dorsal interosseous (FDI) muscle was performed by an expert and a novice examiner, twice in ten healthy volunteers and once in ten tumor patients. The distances between the centers-of-gravity and hotspots were calculated, as were coefficients of variation. This study also compared orthogonal versus variable orientation of the stimulation coil. The mean (range) distance between centers-of-gravity for the expert examiner in the test-retest protocol with healthy volunteers was 4.40 (1.86-7.68) mm. The mean (range) distance between centers-of-gravity for the expert vs. novice examiner was 4.89 (2.39-9.22) mm. There were no significant differences in this result between healthy volunteers and tumor patients. nTMS is sufficiently reliable for clinical use, but examiners should make efforts to minimize sources of error. The reliability of nTMS in tumor patients appears comparable to healthy subjects.


A systematic review and meta-analysis of traumatic intracranial hemorrhage in patients taking prehospital antiplatelet therapy: Is there a role for platelet transfusions?
Platelet transfusion has been utilized to reverse platelet dysfunction in patients on preinjury antiplatelets who have sustained a traumatic intracranial hemorrhage (tICH); however, there is little evidence to substantiate this practice. The objective of this study was to perform a systematic review on the impact of platelet transfusion on survival, hemorrhage progression and need for neurosurgical intervention in patients with tICH on prehospital antiplatelet medication. Controlled, observational and randomized, prospective and retrospective studies describing tICH, preinjury antiplatelet use, and platelet transfusion reported in PubMed, Embase, Cochrane Reviews, Cochrane Trials and Cochrane DARE databases between January 1987 and March 2019 were included. Investigations of concomitant anticoagulant use were excluded. Risk of bias was assessed using the Newcastle-Ottawa scale. We calculated pooled estimates of relative effect of platelet transfusion on the risk of death, hemorrhage progression and need for neurosurgical intervention using the methods of Dersimonian-Laird random-effects meta-analysis. Sensitivity analysis established whether study size contributed to heterogeneity. Subgroup analyses determined whether antiplatelet type, additional blood products/reversal agents, or platelet function assays impacted effect size using meta-regression. Twelve of 18,609 screened references were applicable to our questions and were qualitatively and quantitatively analyzed. We found no association between platelet transfusion and the risk of death in patients with tICH taking prehospital antiplatelets (odds ratio [OR], 1.29; 95% confidence interval [CI], 0.76-2.18; p = 0.346; I = 32.5%). There was no significant reduction in hemorrhage progression (OR, 0.88; 95% CI, 0.34-2.28; p = 0.788; I = 78.1%). There was no significant reduction in the need for neurosurgical intervention (OR, 1.00; 95% CI, 0.53-1.90, p = 0.996; I = 59.1%; p = 0.032). Current evidence does not support the use of platelet transfusion in patients with tICH on prehospital antiplatelets, highlighting the need for a prospective evaluation of this practice. Systematic Reviews and Meta-Analyses, Level III.


An electrogenic component of resting potential in rabbit ventricular muscle?
The resting potential and the intracellular Na and K concentrations (Nai, Ki) were determined at several extracellular K concentrations (Ko) between 0.5 and 18 mM and after inhibition of the sodium pump with 0.5 microM ouabain. Exposure to low Ko (0.5 mM) produced a transient hyperpolarization (from -80 to -100 mV) followed by a depolarization that led to a stable potential of -60 mV within 25 min. Similar potential levels were observed in the presence of ouabain regardless of the Ko/Ki ratio. Intracellular sodium increased at Ko < 5 mM, whereas Ki rose at Ko less than or equal to 1.0 mM. Because of the large decrease of Ki at Ko = 0.5 mM, Ko/Ki was the same at 0.5 and 1 mM. However, the resting potentials at the steady state differed by 50 mV at these concentrations. A PNa/PK of 0.032 for the control conditions was obtained with the Mullins-Noda equation using 2.5 as the Na-K coupling ratio. This PNa/PK value yielded a Goldman potential of -69 mV; so we estimated that electrogenic sodium extrusion contributed -10 mV to the resting potential. The size of the electrogenic potential increased as Ko was lowered from 5 to 1 mM. This finding suggests that the control of the Na-K coupling ratio may be independent of the mechanism that controls the pumping rate.


Energetics of high-speed running: integrating classical theory and contemporary observations.
We hypothesized that the anaerobic power and aerobic power outputs during all-out runs of any common duration between 10 and 150 s would be proportional to the maximum anaerobic (E(an-max)) and aerobic powers (E(aer-max)) available to the individual runner. Seventeen runners who differed in E(an-max) and E(aer-max) (5 sprinters, 5 middle-distance runners, and 7 long distance runners) were tested during treadmill running on a 4.6 degrees incline. E(an-max) was estimated from the fastest treadmill speed subjects could attain for eight steps. E(aer-max) was determined from a progressive, discontinuous, treadmill test to failure. Oxygen deficits and rates of uptake were measured to assess the respective anaerobic and aerobic power outputs during 11-16 all-out treadmill runs that elicited failure between 10 and 220 s. We found that, during all-out runs of any common duration, the relative anaerobic and aerobic powers utilized were largely the same for sprint, middle-distance, and long-distance subjects. The similar fractional utilization of the E(an-max) and E(aer-max) available during high-speed running 1) provides empirical values that modify and advance classic theory, 2) allows rates of anaerobic and aerobic energy release to be quantified from individual maxima and run durations, and 3) explains why the high-speed running performances of different event specialists can be accurately predicted (R(2) = 0.97; n = 254) from two direct measurements and the same exponential time constant.


Root coverage stability: A systematic overview of controlled clinical trials with at least 5 years of follow-up.
To systematically assess the long-term outcome (≥5 years) of root coverage procedures reported in controlled clinical trials. Literature search was performed according to the PRISMA guidelines with the following eligibility criteria: (a) English or German language; (b) controlled (CT) or randomised controlled clinical trials (RCT); (c) root coverage procedure with ≥5 years follow-up; and (d) clinical treatment effect size and/or patient-related outcome measures (PROMs) reported. Four CT and 14 RCT with a follow-up of 5-20 years fulfilled the eligibility criteria; sample size per study ranged from 8 to 70 patients contributing with 18-149 sites. Coronally advanced flap (CAF) and CAF + connective tissue graft (CTG) were the prevalent treatments (i.e., in 24 and 38% of the groups, respectively), while other flap designs and adjuncts (i.e., enamel matrix derivative, bone graft, collagen membrane) were represented only once. For single Miller class I/II gingival recessions (GR), CAF + CTG appeared advantageous compared to other techniques, and provided low residual recession depths (i.e., ≤0.5 mm), and complete root coverage in ≥2/3 of the patients; similar tendency was observed for multiple GR. No data on Miller class III/IV GR is available. No meta-analysis was feasible due to lack of similarity in the clinical and methodological characteristics across the trials and observed comparisons of interventions. CAF + CTG appears to be the 'gold standard' technique for the treatment of single and multiple Miller class I/II GR also in regard to long-term (i.e., ≥5 years of follow-up) treatment outcomes. There is little information regarding the performance, on the long-term, of other techniques and adjuncts.


If You Host It Online, Who Will (and Will Not) Come? Individual and Partner Enrollment in a Web-Based Intervention for Distressed Couples.
Given the increasing utilization of online recruitment and delivery for prevention programming, the current study was designed to examine the ways in which recruitment and eligibility factors affect the resulting size and composition of participants in an online intervention. Study hypotheses were tested from a sample of 2512 low-income individuals who sought to enroll in OurRelationship, a web-based intervention for distressed couples. Results indicated that more than half of the sample (62%) learned about the OurRelationship program from results of an online search engine. Differences in participant characteristics were observed on the basis of recruitment source, with individuals recruited from an online search and from social media being characterized by higher levels of relationship distress and personal psychological distress relative to those who learned about the program through other means. Partner participation requirements also had a significant effect on the final sample of participants, as more than half of help-seeking individuals (52%) had partners who did not complete the screening enrollment form and were thus ineligible to receive services. Furthermore, compared with individuals whose partners completed the enrollment form, individuals whose partners did not participate were characterized by greater levels of break-up potential, physical aggression, communication conflict, psychological distress, and anger. Findings from the study suggest that some, but not all, online sources recruit more at-risk populations as well as illustrate the ways in which partner participation requirements can screen out interested individuals that appear in most need of services. Implications for prevention researchers and practitioners are discussed.


Prognostic role of nm23 gene expression in patients with ovarian cancer.
The aim of the study was to define the prognostic role of the metastasis suppressor gene, nm23, in 50 patients with primary ovarian cancer. Immunohistochemical analysis was performed on formalin-fixed, paraffin-embedded specimens by the primary nm23 monoclonal antibody (Novocastra, NCL-nm23 clone 37.6). Forty-two specimens (84%) showed a positive nm23 staining. The nm23 staining was more intensive in patients with normal serum CA19.9 levels, patients with nonrecurrent disease, and alive patients (p < 0.05). Nm23 expression did not correlate with common clinicopathologic parameters such as histology, grade of differentiation, International Federation of Gynecology and Obstetrics stage, and CA-125. Although the difference was not statistically significant (p = 0.11), we found that nm23 may have a favorable prognostic factor in ovarian cancer. To clarify this subject further, prospective studies on a larger population are needed.


Mouse papillomavirus infections spread to cutaneous sites with progression to malignancy.
We report secondary cutaneous infections in the mouse papillomavirus (MmuPV1)/mouse model. Our previous study demonstrated that cutaneous MmuPV1 infection could spread to mucosal sites. Recently, we observed that mucosal infections could also spread to various cutaneous sites including the back, tail, muzzle and mammary tissues. The secondary site lesions were positive for viral DNA, viral capsid protein and viral particles as determined by in situ hybridization, immunohistochemistry and transmission electron microscopy analyses, respectively. We also demonstrated differential viral production and tumour growth at different secondarily infected skin sites. For example, fewer viral particles were detected in the least susceptible back tissues when compared with those in the infected muzzle and tail, although similar amounts of viral DNA were detected. Follow-up studies demonstrated that significantly lower amounts of viral DNA were packaged in the back lesions. Lavages harvested from the oral cavity and lower genital tracts were equally infectious at both cutaneous and mucosal sites, supporting the broad tissue tropism of this papillomavirus. Importantly, two secondary skin lesions on the forearms of two mice displayed a malignant phenotype at about 9.5 months post-primary infection. Therefore, MmuPV1 induces not only dysplasia at mucosal sites such as the vagina, anus and oral cavity but also skin carcinoma at cutaneous sites. These findings demonstrate that MmuPV1 mucosal infection can be spread to cutaneous sites and suggest that the model could serve a useful role in the study of the viral life cycle and pathogenesis of papillomavirus.


Protean manifestations of vitamin D deficiency, part 3: association with cardiovascular disease and disorders of the central and peripheral nervous systems.
Vitamin D deficiency is associated with the risk factors of inflammation, insulin resistance and endothelial dysfunction, and left ventricular hypertrophy. As a result there is an increase in cardiovascular events (CVEs) associated with vitamin D deficiency. Vitamin D deficiency itself or secondary hyperparathyroidism or both may be responsible for the increase in CVEs. Correction of vitamin D deficiency may decrease the incidence of CVEs. Vitamin D deficiency is also associated with Alzheimer disease, schizophrenia, depression, and chronic pain and muscle weakness. Vitamin D deficiency is early treated with oral vitamin D supplements which may improve the manifestations of the diseases associated with vitamin D deficiency.


Platelet-derived growth factor (PDGF)-PDGF receptor interaction activates bone marrow-derived mesenchymal stromal cells derived from chronic lymphocytic leukemia: implications for an angiogenic switch.
Malignant cells are capable of influencing the microenvironment in a manner that facilitates tumor cell survival. Bidirectional crosstalk between chronic lymphocytic leukemic (CLL) cells and marrow-derived mesenchymal stromal cells (MSCs) activates both cell types. In this study, we observed that the conditioned medium (CM) obtained from CLL cells was able to induce Akt activation in MSC. Subsequent studies investigated the mechanism of MSC activation mediated by CLL-CM. Platelet-derived growth factor receptors (PDGFRs) were selectively activated in MSCs by CLL-CM and found to be critical receptors for CLL-CM-driven MSC proliferation and MSC Akt activation. The known ligands of PDGFR, platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF), were detected in CLL-CM, but PDGF was the predominant ligand involved in the CM-mediated PDGFR activation. Both PDGF and VEGF were found to be elevated in the plasma of CLL patients with a positive association for high-risk factors and more advanced stage. Finally, we demonstrated that PDGF induced MSC VEGF production through a phosphatidylinositol 3-kinase (PI3K)-dependent mechanism. These results show that PDGF-PDGFR signaling influences at least the MSC in the microenvironment of CLL and may play a role in the induction of an angiogenic switch known to be permissive for disease progression.


Protective effects of an interaction between vagus nerve and melatonin on gastric ischemia/reperfusion: the role of oxidative stress.
Vagal pathways in gastrointestinal tract are the most important pathways that regulate ischemia/reperfusion (I/R). Gastrointestinal tract is one of the important sources of melatonin production. The aim of this study was to investigate probable protective effect of the interaction between vagus nerve and melatonin after I/R. This study was performed in male rats that were divided into six groups. Cervical vagus nerve was cut bilaterally after induction of I/R and the right one was stimulated by stimulator. Melatonin or vehicle was injected intraperitoneally. The stomach was removed for histopathological and biochemical investigations. A significant decrease in infiltration of gastric neutrophils and malondialdehyde (MDA) level after I/R was induced by melatonin and was disappeared after vagotomy. The stimulation of vagus nerve significantly enhanced these effects of melatonin. However, a stimulation of vagus nerve alone increased neutrophils infiltration and MDA level. Melatonin significantly increased the activities of catalase, glutathione peroxidase (GPx), superoxide dismutases (SOD). Unlike stimulation of vagus nerve, vagotomy decreased these effects of melatonin. According to these results, it is probable that protective effects of melatonin after I/R may be mediated by vagus nerve. Therefore, there is an interaction between melatonin and vagus nerve in their protective effects.


The reductive half-reaction of xanthine dehydrogenase from Rhodobacter capsulatus: the role of Glu232 in catalysis.
The kinetic properties of an E232Q variant of the xanthine dehydrogenase from Rhodobacter capsulatus have been examined to ascertain whether Glu(232) in wild-type enzyme is protonated or unprotonated in the course of catalysis at neutral pH. We find that kred, the limiting rate constant for reduction at high [xanthine], is significantly compromised in the variant, a result that is inconsistent with Glu(232) being neutral in the active site of the wild-type enzyme. A comparison of the pH dependence of both kred and kred/Kd from reductive half-reaction experiments between wild-type and enzyme and the E232Q variant suggests that the ionized Glu(232) of wild-type enzyme plays an important role in catalysis by discriminating against the monoanionic form of substrate, effectively increasing the pKa of substrate by two pH units and ensuring that at physiological pH the neutral form of substrate predominates in the Michaelis complex. A kinetic isotope study of the wild-type R. capsulatus enzyme indicates that, as previously determined for the bovine and chicken enzymes, product release is principally rate-limiting in catalysis. The disparity in rate constants for the chemical step of the reaction and product release, however, is not as great in the bacterial enzyme as compared with the vertebrate forms. The results indicate that the bacterial and bovine enzymes catalyze the chemical step of the reaction to the same degree and that the faster turnover observed with the bacterial enzyme is due to a faster rate constant for product release than is seen with the vertebrate enzyme.


P2 purinoceptor of the globular substance in the otoconial membrane of the guinea pig inner ear.
The biological characteristics of the globular substance, a precursor of otoconia, are unclear. In the present study, the ATP-induced internal free Ca2+ concentration ([Ca2+]i) changes of the globular substance and the ATP distribution in the vestibular organ were investigated using a Ca2+ indicator, fluo 3, and an adenine nucleotide-specific fluorochrome, quinacrine, by means of confocal laser scanning microscopy. [Ca2+]i showed a rapid and dose-dependent increase in response to ATP with a 50% effective concentration (EC50) of 16.7 microM. This reaction was independent of external Ca2+, indicating the presence of an internal Ca2+ reservoir. Neither adenosine, alpha, beta-methylene-ATP, 3'-O-(4-benzoylbenzoyl)-ATP, ADP, nor UTP evoked this reaction, whereas 2-methylthio-ATP induced an increase of [Ca2+]i with an EC50 of 14.4 microM. Moreover, P2 antagonists, reactive blue 2 and suramin, and a phospholipase C inhibitor, U-73122, inhibited the ATP-induced [Ca2+]i increase. These findings indicate the presence of a P2Y purinoceptor on the globular substance. In addition, granular fluorescence was observed in the quinacrine-stained macular sensory epithelium, indicating the presence of ATP-containing granules in this tissue. These results suggest that a paracrine mechanism involving ATP may exist in the macula and that this mechanism regulates the biological behavior of the globular substance.


Energetic and Metabolic Power Demands of National Rugby League Match-Play.
The purpose of this study was to apply a time-motion model to estimate and describe the energy expenditure and metabolic power demands of playing positions in elite rugby league match-play, utilizing Global Positioning System (GPS) devices. 18 elite rugby league players participated in this study. Players' positional groups included: outside backs (n=59 files, n=4 players), adjustables (n=74 files, n=4 players), wide-running (n=104 files, n=7 players) and hit-up forwards (n=36 files, n=3 players). Outside backs expended the greatest total energy (40.1±5.0 kJ·kg(-1)) per match, equivalent to 8.1%, 26.6% and 61.9% greater energy than adjustables, wide-running and hit-up forwards, respectively. Adjustables attained an anaerobic index 7.3% higher than wide-running forwards, 19.7% higher than hit-up forwards (p=0.001) and 43.2% higher than outside backs (p<0.001). Wide-running forwards achieved an anaerobic index (0.34±0.04) 11% and 32.8% higher than hit-up forwards (p=0.001) and outside backs (p<0.001), respectively. Mean power of adjustables (10.0±0.9 W·kg(-1)) was significantly higher than all other groups (outside backs: 28.8%, 7.8±1.0; hit-up: 12.4%, 8.9±0.6; and wide-running: 8.7%, 9.2±0.7 forwards) (p<0.001). Energetics indices indicated differing metabolic demands for all positional groups, suggesting position-specific conditioning drills are required to replicate the energetic demands of match-play.


Poland's syndrome with lung cancer. A case report.
Poland's syndrome, a rare congenital anomaly characterized by pectoralis muscle defect, has been reported in association with lymphoreticular malignancies and some solid tumors. Lung cancer associated with Poland's syndrome has not been previously described. We present the first report of a case of Poland's syndrome associated with lung cancer and demonstrate the CT findings.


The effects of exercise modality and intensity on energy expenditure and cardiorespiratory response in adults with obesity and treated obstructive sleep apnoea.
To inform recommendations for the exercise component of a healthy lifestyle intervention for adults with obesity and treated obstructive sleep apnoea (OSA), we investigated the total energy expenditure (EE) and cardiorespiratory response to weight-supported (cycling) and unsupported (walking) exercise. Individuals with treated OSA and a body mass index (BMI) > 30 kg/m2 performed an incremental cardiopulmonary exercise test on a cycle ergometer and a treadmill to determine the peak oxygen uptake [Formula: see text]. Participants subsequently completed two endurance tests on each modality, matched at 80% and 60% of the highest [Formula: see text] determined by the incremental tests, to intolerance. The cardiorespiratory response was measured and total EE was estimated from the [Formula: see text]. Sixteen participants completed all six tests: mean [SD] age 57 [13] years and median [IQ range] BMI 33.3 [30.8-35.3] kg/m2. Total EE during treadmill walking was greater than cycling at both high (158 [101] vs. 29 [15] kcal; p < 0.001) and moderate (178 [100] vs. 85 [59] kcal; p = 0.002) intensities, respectively, with similar cardiorespiratory responses and pattern of EE during rest, exercise and recovery. Contrary to current guidelines, walking might be the preferred training modality to achieve the combination of weight loss and increased cardiorespiratory fitness in adults with obesity and treated OSA.


Characteristics of the adenylyl cyclase system of differentiating rabbit bone marrow erythroblasts.
Changes in the cellular content of cyclic AMP and in the activities of adenylyl cyclase, cyclic AMP phosphodiesterase and cyclic AMP-dependent protein kinases during differentiation of rabbit bone marrow erythroid cells were investigated. The cells were separated by velocity sedimentation at unit gravity into six fractions corresponding to different stages of development: proerythroblasts, basophilic erythroblasts, polychromatic cells, early orthochromatic and late orthochromatic cells and reticulocytes. Adenylyl cyclase activity was found to decrease continuously as the cells developed, from approx. 180 pmoles cyclic AMP formed/mg of protein/20 min in proerythroblasts to 10 pmoles in circulating reticulocytes. The proerythroblasts were the richest cells in cyclic AMP which is present at a cellular concentration of approx. 1.4 microM. In basophilic cells the cyclic AMP content was about 80% lower than in proerythroblasts. No further changes in cyclic AMP levels were observed after the final cell division. Cyclic AMP phosphodiesterase was found to be very active in the most immature cells, the proerythroblasts. After differentiation into basophilic erythroblasts, a 4-fold decrease in cyclic AMP phosphodiesterase activity occurred. In polychromatic cells there was a further drop in phosphodiesterase activity and after the last cell division the enzyme activity was constant and very low. Both cytosolic cyclic AMP-binding capacity and cytosolic cyclic AMP-dependent protein kinase activity decreased in dividing rabbit bone marrow erythroblasts when calculated in terms of cell number but remained constant per cell volume. After the final cell division, cyclic AMP-dependent protein kinase activity did not change further, whereas cyclic AMP-binding capacity declined. There were no qualitative but only quantitative changes in the cyclic AMP-binding proteins that are present in the cytosol of developing erythroblasts. In the immature cells, the apparent Kd for the interaction of binding proteins with cyclic AMP was 4 . 10(-8) M. The data suggest that changes in cyclic AMP-binding activity during differentiation of erythroid cells are due both to changes in the amount of binding proteins and their affinity for cyclic AMP. The phosphorylation of rabbit erythroblast plasma membrane proteins by membrane-associated protein kinase(s) was found to be cyclic AMP-dependent in dividing cells during the early stages of differentiation. When the erythroid cells reach the non-dividing stage in their development, autophosphorylation of membrane ghosts was no longer stimulated by cyclic AMP.(ABSTRACT TRUNCATED AT 400 WORDS)


Simultaneous determination of valsartan, amlodipine besylate and hydrochlorothiazide using capillary zone electrophoresis (CZE).
A capillary zone electrophoresis method was developed for the simultaneous determination of valsartan (VAL), amlodipine besylate (AML) and hydrochlorothiazide (HCZ) in their combined tablets. Separation was achieved on a fused silica capillary by applying a potential of 15 kV (positive polarity) and a running background electrolyte containing 40 mM phosphate buffer at pH 7.5 with UV detection at 230 nm. The samples were injected hydrodynamically for 3s at 0.5 psi and the temperature of the capillary cartridge was kept at 25 degrees C. Pyrazinoic acid was used as an internal standard. The method was validated according to ICH guidelines regarding specificity, linearity, limits of detection and quantitation, accuracy and precision, (Supplementary materials, Table S2). The method showed satisfactory linearity in the ranges of 10-200, 2-20 and 2-20 μg mL(-1) with LODs of 1.82, 0.39, 0.65 μg mL(-1) and LOQs of 5.51, 1.17, 1.96 μg mL(-1) for VAL, AML and HCZ, respectively. The proposed method was successfully applied for the analysis of the studied drugs in their laboratory prepared mixtures and co-formulated tablets. The results were compared with reported methods and no significant differences were found. The proposed method can be used for quality control of the cited drugs in ordinary laboratories.


Oral manifestations of HIV infection in relation to clinical and CD4 immunological status in northern and southern Thai patients.
To assess prevalence and variations in the oral manifestations of HIV in HIV-infected subjects in southern Thailand (a new HIV epidemic) and northern Thailand (a mature epidemic), and the association with age, sex, risk behaviours, CD4 count and medication used. A total of 102 and 135 HIV-infected individuals were enrolled in northern and southern hospitals, respectively. Oral and haematological examination was performed after sociodemographic interview of the patients. Clinical history was retrieved from patients' medical records. Oral candidiasis (OC, 55%), oral hairy leucoplakia (OHL, 21%) and HIV-associated-periodontal disease (14%) were among the most common oral lesions in southern Thailand. OHL (38%), OC (25%), HIV-associated-periodontal disease (15%) were the three most common lesions in the north. A significant association was found between any oral lesion, OC, particularly the pseudomembranous type (PC), and CD4 < 200 cells mm(-3) at both sites. A negative relationship was found between systemic antifungal treatment and OC including PC and erythematous candidiasis (EC) in the southern data. OHL showed a positive relationship with male sex and a negative relationship with antiretroviral treatment in the northern site. Younger age and being a current smoker were positively associated with oral lesions in the southern group. OC, particularly PC, could be useful as a marker for immunosuppression, particularly where CD4 count cannot be determined routinely. Antifungal treatment is of benefit in the subjects who cannot afford highly active antiretroviral therapy (HAART).


Whole genome detection of signature of positive selection in African cattle reveals selection for thermotolerance.
As African indigenous cattle evolved in a hot tropical climate, they have developed an inherent thermotolerance; survival mechanisms include a light-colored and shiny coat, increased sweating, and cellular and molecular mechanisms to cope with high environmental temperature. Here, we report the positive selection signature of genes in African cattle breeds which contribute for their heat tolerance mechanisms. We compared the genomes of five indigenous African cattle breeds with the genomes of four commercial cattle breeds using cross-population composite likelihood ratio (XP-CLR) and cross-population extended haplotype homozygosity (XP-EHH) statistical methods. We identified 296 (XP-EHH) and 327 (XP-CLR) positively selected genes. Gene ontology analysis resulted in 41 biological process terms and six Kyoto Encyclopedia of Genes and Genomes pathways. Several genes and pathways were found to be involved in oxidative stress response, osmotic stress response, heat shock response, hair and skin properties, sweat gland development and sweating, feed intake and metabolism, and reproduction functions. The genes and pathways identified directly or indirectly contribute to the superior heat tolerance mechanisms in African cattle populations. The result will improve our understanding of the biological mechanisms of heat tolerance in African cattle breeds and opens an avenue for further study.


Stem cell therapies for the lysosomal storage diseases - the quintessential neurodegenerative diseases.
As a novel neurotherapeutic strategy, stem cell transplantation has received considerable attention. However, little focus of this attention has been devoted to the probabilities of success of stem cell therapies for specific neurological disorders. Given the complexities of the cellular organization of the nervous system and the manner in which it is assembled during development, it seems unlikely that a cellular replacement strategy will succeed for any but the simplest of neurological disorders in the near future. A general strategy for stem cell transplantation to prevent or minimize neurological disorders is much more likely to succeed. The lysosomal storage diseases represent the quintessential neurodegenerative diseases for which preventative stem cell transplantation will both likely succeed and set the stage for therapeutic approaches to other neurodegenerative diseases.


NF-κB and the cell cycle.
The NF-κB (nuclear factor κB) transcription factor family is a pleiotropic regulator of many cellular pathways, providing a mechanism for the cell to respond to a wide variety of stimuli and environmental challenges. It is not surprising therefore that an important component of NF-κB's function includes regulation of the cell cycle. However, this aspect of its behaviour is often overlooked and receives less attention than its ability to induce inflammatory gene expression. In the present article, we provide an updated review of the current state of our knowledge about integration of NF-κB activity with cell cycle regulation, including newly characterized direct and indirect target genes in addition to the mechanisms through which NF-κB itself can be regulated by the cell cycle.


Shear banding in monodisperse polymer melt.
We performed a series of molecular dynamics simulations on monodisperse polymer melts to investigate the formation of shear banding. Under high shear rates, shear banding occurs, which is intimately accompanied by the entanglement heterogeneity. Interestingly, the same linear relationship between the end-to-end distance Ree and entanglement density Z is observed at homogeneous flow before the onset of shear banding and at the shear banding state, where Ree ∼ ln(Wi0.87)-ξ0Z is proposed as the criterion to describe the dynamic force balance of the molecular chain in flow with a high rate. Deviating from this relation leads to a force imbalance and results in the emergence of shear banding. We establish a scaling relation between the disentanglement rate Vd and the Weissenberg number Wi as Vd∼Wi0.87 for stable flow in homogeneous shear and shear banding states. The formation of shear banding prevents chains from further stretching and disentanglement. The transition from homogeneous shear to shear banding partially dissipates the increased free energy from shear and reduces the free energy of the system.


[Activity of alpha-chymotrypsin immobilized in nanocapsules of poly(N,N-didodecyl-N,N-diallylammonium bromide)].
Biocatalytic nanocapsule systems containing alpha-chymotrypsin in the inner aqueous cavity were synthesized. These systems can function in both organic solvents and aqueous media. To this end, the reversed hydrated micelles of N,N-diallyl-N,N-didodecylammonium bromide (DDAB) in cyclohexane (w0 = 22) with alpha-chymotrypsin encapsulated were subjected to UV-induced polymerization. After precipitation with acetone, the nanocapsules were transferred into water with the use of AOT and subsequent sonication. The unilamellar liposomes thus formed have an inner monolayer from the polyDDAB network, an outer monolayer from the AOT molecules, and contain alpha-chymotrypsin in the inner aqueous medium. According to the light scattering data, the mean outer diameter of the nanocapsules is 20 nm. When compared with free enzyme in respect to p-nitrophenyl acetate hydrolysis, the alpha-chymotrypsin encapsulated in the liposomes exhibits the same K(m) value, but its values of kcat and of the inhibition constant by the reaction product Kp decreased by factors of 1.2 and 1.6, respectively. The alpha-chymotrypsin immobilized in the nanocapsules is thermally stable and catalytically active up to 80 degrees C. The polymer network hinders conformational rearrangements of the enzyme molecule upon heating.


Fear appeal effectiveness in antismoking campaigns: do anger and smoking matter?
Although fear appeals are widely used in health campaigns, empirical studies have produced conflicting results regarding their effectiveness. To understand the impact of fear appeal in health campaigns, this study investigates the relationship between fear, anger, and smoking status in the context of an antismoking campaign. Using a survey of 829 people in South Korea, this study tests the three-way interactive model with the PROCESS macro. Results indicate that the effect of fear on attitudes toward the campaign depends on the level of anger, and show that the impact of 'anger-adjusted' additional fear appears only among nonsmokers. Implications for antismoking advertising strategies are discussed.


Catalytic ferrous iron in amniotic fluid as a predictive marker of human maternal-fetal disorders.
Amniotic fluid contains numerous biomolecules derived from fetus and mother, thus providing precious information on pregnancy. Here, we evaluated oxidative stress of human amniotic fluid and measured the concentration of catalytic Fe(II). Amniotic fluid samples were collected with consent from a total of 89 subjects in Nagoya University Hospital, under necessary medical interventions: normal pregnancy at term, normal pregnancy at the 2nd trimester, preterm delivery with maternal disorders but without fetal disorders, congenital diaphragmatic hernia, fetal growth restriction, pregnancy-induced hypertension, gestational diabetes mellitus, Down syndrome and trisomy 18. Catalytic Fe(II) and oxidative stress markers (8-hydroxy-2'-deoxyguanosine, 8-OHdG; dityrosine) were determined with RhoNox-1 and specific antibodies, respectively, using plate assays. Levels of 8-OHdG and dityrosine were higher in the 3rd trimester compared with the 2nd trimester in normal subjects, and the abnormal groups generally showed lower levels than the controls, thus suggesting that they represent fetal metabolic activities. In contrast, catalytic Fe(II) was higher in the 2nd trimester than the 3rd trimester in the normal subjects, and overall the abnormal groups showed higher levels than the controls, suggesting that high catalytic Fe(II) at late gestation reflects fetal pathologic alterations. Notably, products of H2O2 and catalytic Fe(II) remained almost constant in amniotic fluid.


Global prevalence of ankylosing spondylitis.
For effective health care provision, knowledge of disease prevalence is paramount. There has been no systematic endeavour to establish continent-based AS estimates, however, prevalence is thought to vary by country and background HLA-B27 prevalence. This study aimed to estimate AS prevalence worldwide and to calculate the expected number of cases. A systematic literature search was conducted. Prevalence data were extracted and used to calculate the mean prevalence by continent and the expected number of cases based on country-specific prevalence (or, if missing, the prevalence from neighbouring countries). A second estimate was made using the prevalence from countries with similar HLA-B27 prevalences if a country-specific prevalence estimate was not available. The mean AS prevalence per 10,000 (from 36 eligible studies) was 23.8 in Europe, 16.7 in Asia, 31.9 in North America, 10.2 in Latin America and 7.4 in Africa. Additional estimates, weighted by study size, were calculated as 18.6, 18.0 and 12.2 for Europe, Asia and Latin America, respectively. There were sufficient studies to estimate the number of cases in Europe and Asia, calculated to be 1.30-1.56 million and 4.63-4.98 million, respectively. This study represents the first systematic attempt to collate estimates of AS prevalence into a single continent-based estimate. In addition, the number of expected cases in Europe and Asia was estimated. Through reviewing the current literature, it is apparent that the continuing conduct of epidemiological studies of AS prevalence is of great importance, particularly as diagnostic capabilities improve and with the recent development of the criteria for axial SpA.


Self-absorption correction for carbon-14.
A new and simple technique has been developed (10) for correcting C(14) radioactivity measurements for loss of radiation due to self-absorption. Results are discussed which show the applicability of the technique for many different counting situations. It is found that the absorption of radiation follows a hyperbolic law much more closely than an exponential one. Furthermore, the absorption coefficient for the sample itself has been found to be a function of weight rather than a constant, as had been assumed for the derivation of the law of exponential absorption.


Bias for consonantal information over vocalic information in 30-month-olds: cross-linguistic evidence from French and English.
Using a name-based categorization task, Nazzi found in 2005 that French-learning 20-month-olds can make use of one-feature consonantal contrasts between new labels but fail to do so with one-feature vocalic contrasts. This asymmetry was interpreted as developmental evidence for the proposal that consonants play a more important role than vowels at the lexical level. In the current study using the same task, we first show that by 30 months French-learning infants can make use of one-feature vocalic contrasts (e.g., /pize/-/pyze/). Second, we show that in a situation where infants must neglect either a consonantal one-feature change or a vocalic one-feature change (e.g., match a /pide/ with either a /tide/ or a /pyde/), both French- and English-learning 30-month-olds choose to neglect the vocalic change rather than the consonantal change. We argue that these results suggest that by 30 months of age, infants still give less weight to vocalic information than to consonantal information in a lexically related task even though they are able to process fine vocalic information.


Physiological concentrations of DHEA in human hair.
In 1974, steroids were added to the list of doping agents banned by the International Olympic Committee because of their effects on the performance of the athletes. Dehydroepiandrosterone (DHEA) is a steroid hormone naturally produced by the adrenal glands and by the ovaries. DHEA can be converted into other hormones, including estrogen and testosterone. In the United States, DHEA is classified as a nutritional supplement. This is not the case in France, where the drug is listed as a doping agent. As athletes can abuse DHEA to benefit from its conversion to testosterone, there is a need to establish the physiological range of DHEA concentrations in human hair. DHEA was investigated in hair obtained from 27 control subjects, including 15 males and 12 females aged 17-42 years. After decontamination with dichloromethane, 100 mg of hair was incubated in 1 M NaOH in presence of 1 ng of testosterone-d3. After neutralization, the extract was purified using solid-phase extraction with Isolute C18 columns and subsequent liquid-liquid extraction with pentane. After silylation, DHEA was analyzed by gas chromatography-mass spectrometry. Results were linear in the range 1-20 pg/mg. Relative extraction recovery was 91.6% with a limit of detection of 0.5 pg/mg. Concentrations were in the range 1.2-6.7 pg/mg (mean value of 4.3 pg/mg) and 0.5 to 10.6 pg/mg (mean value of 5.3 pg/mg) for the males and females, respectively. Extensive chromatographic procedures (two purification steps by solid-phase and liquid-liquid extraction, combined with injection of 4 microL through the column in pulsed mode) were analytical prerequisites for successful identification of DHEA in hair because of the low target concentrations. This new technology may find useful applications in anabolic abuse control.


Pellagra encephalopathy among tuberculous patients: its relation to isoniazid therapy.
Eight cases of pellagra, diagnosed on the grounds of neuropathological findings and retrospective study of clinical data, were found among 106 necropsy cases of tuberculosis. Although these eight patients had shown various mental, neurological and gastrointestinal symptoms, as well as skin lesions, the diagnosis of pellagra had not been made clinically. In all the patients, pellagra symptoms appeared during isoniazid therapy. Death occurred 4 to 16 weeks later. Isoniazid inhibits the conversion of tryptophan to niacin and may induce pellagra, particularly in poorly nourished patients. Pellagra should be suspected whenever tuberculous patients under treatment with isoniazid develop mental, neurological or gastrointestinal symptoms, even in the absence of typical pellagra dermatitis.


Exchange coupling torque in ferrimagnetic Co/Gd bilayer maximized near angular momentum compensation temperature.
Highly efficient current-induced motion of chiral domain walls was recently demonstrated in synthetic antiferromagnetic (SAF) structures due to an exchange coupling torque (ECT). The ECT derives from the antiferromagnetic exchange coupling through a ruthenium spacer layer between the two perpendicularly magnetized layers that comprise the SAF. Here we report that the same ECT mechanism applies to ferrimagnetic bi-layers formed from adjacent Co and Gd layers. In particular, we show that the ECT is maximized at the temperature TA where the Co and Gd angular momenta balance each other, rather than at their magnetization compensation temperature TM. The current induced velocity of the domain walls is highly sensitive to longitudinal magnetic fields but we show that this not the case near TA. Our studies provide new insight into the ECT mechanism for ferrimagnetic systems. The high efficiency of the ECT makes it important for advanced domain wall based spintronic devices.


Activities of enzymes of acetoacetate metabolism in rat brown adipose tissue during development.
The activities of two mitochondrial enzymes concerned in the utilization of acetoacetate, namely 3-oxoacid CoA-transferase and acetoacetyl-CoA thiolase, were high throughout the suckling and weanling period in brown adipose tissue of the rat. In contrast, 3-hydroxybutyrate dehydrogenase activity was comparatively low during this period. The activity of cytosolic acetoacetyl-CoA synthetase (involved in lipogenesis) declined after birth and remained low until the pups were weaned. Experiments with brown-adipose-tissue slices from weanling rats indicated that 70% of the [3-14C]acetoacetate utilized was oxidized to 14CO2, and this value was not altered appreciably by the addition of glucose and insulin.


Metabolic effect of 3,3',5'-triiodothyronine in cultured growth hormone-producing rat pituitary tumor cells. Evidence for a unique mechanism of thyroid hormone action.
Physiologic levels of 3,3',5'-triiodothyronine (rT3) are generally believed to have minimal metabolic effects in the pituitary gland and other tissues. In the present studies, the regulatory role of rT3 and other thyroid hormones on iodothyronine 5'-deiodinase (I5'D) activity was studied in a growth hormone-producing rat pituitary tumor cell line (GH3 cells). I5'D activity was thiol-dependent and displayed nonlinear reaction kinetics suggesting the presence of two enzymatic processes, one having a low Michaelis constant (Km for thyroxine [T4] of 2 nM) and a second with a high Km value (0.9 microM). Growth of cells in hormone-depleted medium resulted in a two- to 3.5-fold increase in low Km I5'D activity (P less than 0.001). The addition of thyroid hormones to the culture medium resulted in a rapid, dose-dependent inhibition of low Km I5'D activity with the following order of analogue potency: rT3 greater than or equal to T4 greater than 3,5,3'-triiodothyronine (T3). Using serum-free culture conditions, rT3 was approximately 50 times more active than T3. These inhibitory effects were noted within 15 min of hormone addition and could not be attributed to substrate competition with T4. These findings suggest that the control of T4 to T3 conversion by thyroid hormones in the anterior pituitary gland is mediated by a unique cellular mechanism that is independent of the nuclear T3 receptor; and under some circumstances, rT3 may play a regulatory role in controlling this enzymatic process.


Hydrogen inhalation inhibits microglia activation and neuroinflammation in a rat model of traumatic brain injury.
Traumatic brain injury (TBI) is a major cause of mortality and disability worldwide. To date, therapies to treat any forms of TBI are still limited. Recent studies have demonstrated the potential neuroprotective effects of molecular hydrogen on TBI. Although it has been demonstrated that hydrogen inhalation (HI) for about 5 hrs immediately after TBI has a beneficial effect on brain injury, the most effective intervention procedure in the treatment of TBI remains unknown. The mechanism underlying the neuroprotective effects of HI on TBI also needs to be further investigated. Our results showed that inhalation of 4% hydrogen during the first day after TBI was the most effective hydrogen intervention procedure in the treatment of TBI. Pathological examination showed that HI could attenuate TBI-induced reactive astrocytosis and microglial activation. Nissl staining demonstrated a significant decrease in the number of nissl-stained dark neurons (N-DNs) in HI group compared to TBI group at 2 h post-TBI, and the TBI-induced neuronal loss was attenuated by HI at day 3 post-TBI. IHC staining showed that HI resulted a decrease in CD16-positive cells and a further increase in CD206-positive cells as compared to TBI group. Multiplex cytokine assay demonstrated the most profound regulatory effects induced by HI on the levels of IL-12, IFN-γ, and GM-CSF at 24 h post-TBI, which confirmed the inhibitory effect of hydrogen on microglia activation. We concluded that inhalation of 4% hydrogen during the first day after TBI was the most effective intervention procedure in the treatment of TBI. Our results also showed that hydrogen may exert its protective effects on TBI via inhibition of microglia activation and neuroinflammation.


Light chain myeloma with features of the adult Fanconi syndrome: six years remission following one course of melphalan.
A 41-year-old woman, presenting with renal failure, renal glucosuria and moderate anemia, was found to have light chain myeloma, indicated by a kappa chain M-component in the serum, heavy urinary excretion of kappa chains and plasma cell infiltration of the bone marrow. After administration of one course of melphalan, resulting in transient pancytopenia, the light chains disappeared completely, renal function returned to normal, glucosuria disappeared and the Hb concentration normalized. During an observation period of six years she has remained in good health and there has been no sign of relapse.


What do parents know about the malformations afflicting the hearts of their children?
Traditionally, medical professionals have entrusted the parents of children with chronic illness with the task of imparting knowledge about the illness to the children. This practice assumes that parents understand the illness, and that they pass on their knowledge in a manner appropriate for the individual child. The aim of our study was to assess the knowledge that parents of children with a cardiac defect have about the malformation in the heart of their child. We sent a modified version of the Leuven questionnaire to 350 families. The questionnaire was filled in and sent back to our centre by 148 families. Parents showed a good knowledge of the name and anatomical characteristics of the cardiac disease suffered by their child. Parents with a child taking drugs were not very knowledgeable about the regime, side effects, and interaction with other drugs or food. Only one-quarter of the parents knew the definition of endocarditis, although almost two-fifths were aware that unexplained fever for more than 5 days was the most typical symptom. Less than half of the parents knew that endocarditis could recur. About two-fifths of parents knew the real possibility of their child being involved in competitive sports; but almost half of parents were unable to answer this question. The poor knowledge about particular aspects of the disease, treatment, and preventive measures revealed by our parents may have major consequences. The results of our study are relevant to general daily clinical practice.


Hybrid light applied with 37% carbamide peroxide bleaching agent with or without titanium dioxide potentializes color change effectiveness.
The effectiveness of dental color change was assessed by incorporating titanium dioxide (TiO2) into 37% carbamide peroxide bleaching agent associated with hybrid light. Fifty bovine incisors were selected to receive the bleaching treatment, and separated into five groups (n = 10): 35% hydrogen peroxide (HP) (Whiteness HP, FGM/HP); 37% carbamide peroxide (CP) (Whiteness SuperEndo, FGM/CP); CP + hybrid light (HL) (CP HL); CP + 1% TiO2 (CP TiO2); CP TiO2 + hybrid light (CP TiO2 HL). The bleaching gels were applied to the dental surface for 30 min. Hybrid light (Whitening Plus, DMC/infrared laser diodes + blue LEDs +violet LEDs) was applied with 1 min of active light, alternating with 1 min of pause. A spectrophotometer (VITA Easyshade® Advance, Vita) was used to determine the color of the dental elements at baseline and time points after the 1st, 2nd and 3rd bleaching sessions. Color change effectiveness was evaluated using Vita Classical, CIEL*a*b*, WID and ΔEab, ΔE00 and ΔWID parameters. Generalized mixed linear models for repeated measures (α = 5%) showed significant decrease in Vita Classical scores and a* and b* values, as well as an increase in L* and ∆WID values for all the groups. Higher color change values for ΔEab were observed for CP HL and CP TiO2 HL, while those of ΔE00 and ΔWID were higher for CP TiO2 HL at the end of the bleaching treatment. Hybrid light applied with TiO2 incorporated into CP potentiated the effectiveness of the color change in the tooth structure.


The effect of the addition of a polyglutamate motif to RGD on peptide tethering to hydroxyapatite and the promotion of mesenchymal stem cell adhesion.
Mimicking endogenous bone-binding proteins, RGD peptides have been synthesized with polyacidic amino acid domains in order to ionically tether the peptides to bone-like synthetic biomaterials, including hydroxyapatite (HA). However, a direct comparison of unmodified RGD with polyacidic-conjugated RGD has not been performed, and thus a benefit for the acidic domain has not been established. We evaluated the peptide/HA bond of RGD peptides with and without an attached polyglutamate sequence (E(7)), as well as examined mesenchymal stem cell (MSC) adhesion and morphology as they were affected by the conjugated peptide. We found that significantly more E(7)RGD was bound to HA than RGD at all coating concentrations tested, and moreover, more E(7)RGD was retained on the HA surface even after extended washing in serum-free media. Consistent with in vitro results, higher levels of E(7)RGD than RGD remained on HA that had been implanted in vivo for 24 h, indicating that the polyacidic domain improved peptide-binding efficiency. At several peptide concentrations, E(7)RGD increased cell adhesion compared to RGD surfaces, establishing a biological benefit for the E(7) modification. In addition, HA pre-coated sequentially with low-density E(7)RGD (1-10 microg/ml) and serum (FBS) stimulated cell adhesion and spreading, compared to either coating alone, suggesting that an ionic linkage allows for the potential adsorption of serum proteins to unoccupied sites, which may be important for bone formation in vivo. Collectively, these results suggest that tethering peptides to HA via a polyglutamate domain is an effective method for improving the peptide/HA bond, as well as for enhancing MSC adhesion.


Cation-induced kinetic heterogeneity of the intron-exon recognition in single group II introns.
RNA is commonly believed to undergo a number of sequential folding steps before reaching its functional fold, i.e., the global minimum in the free energy landscape. However, there is accumulating evidence that several functional conformations are often in coexistence, corresponding to multiple (local) minima in the folding landscape. Here we use the 5'-exon-intron recognition duplex of a self-splicing ribozyme as a model system to study the influence of Mg(2+) and Ca(2+) on RNA tertiary structure formation. Bulk and single-molecule spectroscopy reveal that near-physiological M(2+) concentrations strongly promote interstrand association. Moreover, the presence of M(2+) leads to pronounced kinetic heterogeneity, suggesting the coexistence of multiple docked and undocked RNA conformations. Heterogeneity is found to decrease at saturating M(2+) concentrations. Using NMR, we locate specific Mg(2+) binding pockets and quantify their affinity toward Mg(2+). Mg(2+) pulse experiments show that M(2+) exchange occurs on the timescale of seconds. This unprecedented combination of NMR and single-molecule Förster resonance energy transfer demonstrates for the first time to our knowledge that a rugged free energy landscape coincides with incomplete occupation of specific M(2+) binding sites at near-physiological M(2+) concentrations. Unconventional kinetics in nucleic acid folding frequently encountered in single-molecule experiments are therefore likely to originate from a spectrum of conformations that differ in the occupation of M(2+) binding sites.


Hybridization enables the fixation of selfish queen genotypes in eusocial colonies.
A eusocial colony typically consists of two main castes: queens that reproduce and sterile workers that help them. This division of labor, however, is vulnerable to genetic elements that favor the development of their carriers into queens. Several factors, such as intracolonial relatedness, can modulate the spread of such caste-biasing genotypes. Here we investigate the effects of a notable yet understudied ecological setting: where larvae produced by hybridization develop into sterile workers. Using mathematical modeling, we show that the coevolution of hybridization with caste determination readily triggers an evolutionary arms race between nonhybrid larvae that increasingly develop into queens, and queens that increasingly hybridize to produce workers. Even where hybridization reduces worker function and colony fitness, this race can lead to the loss of developmental plasticity and to genetically hard-wired caste determination. Overall, our results may help understand the repeated evolution toward remarkable reproductive systems (e.g., social hybridogenesis) observed in several ant species.


The impact of ruminative processing on the experience of self-referent intrusive memories in dysphoria.
The current study sought to experimentally assess the differential effects of analytical ruminative processing and distraction on the experience of self-referent naturally occurring intrusive memories in a sample of dysphoric (BDI-II > or = 12) participants. Seventy-seven undergraduate participants completed a memory interview to elicit details about a self-referential intrusion and were randomly assigned to either an analytical rumination or distraction condition. Subsequent to the rumination induction, participants rated their intrusive memory as more negative, more distressing, and more evocative of a negative emotional response compared to participants who were allocated to the distraction induction. Inducing analytical rumination also resulted in participants reporting worse (i.e., more sad) mood relative to those in the distraction condition. The findings align with the suggestion that depressed individuals may get caught up in a ruminative cycle that, due to the documented effects of analytical self-focus, exacerbate the emotional response elicited by intrusions and perpetuate biased attentional focus on them. Directions for future investigations of the cognitive processes that are important in the maintenance of intrusions in depressive disorders are discussed.


Evolution of urea transporters in vertebrates: adaptation to urea's multiple roles and metabolic sources.
In the two decades since the first cloning of the mammalian kidney urea transporter (UT-A), UT genes have been identified in a plethora of organisms, ranging from single-celled bacteria to metazoans. In this review, focusing mainly on vertebrates, we first reiterate the multiple catabolic and anabolic pathways that produce urea, then we reconstruct the phylogenetic history of UTs, and finally we examine the tissue distribution of UTs in selected vertebrate species. Our analysis reveals that from an ancestral UT, three homologues evolved in piscine lineages (UT-A, UT-C and UT-D), followed by a subsequent reduction to a single UT-A in lobe-finned fish and amphibians. A later internal tandem duplication of UT-A occurred in the amniote lineage (UT-A1), followed by a second tandem duplication in mammals to give rise to UT-B. While the expected UT expression is evident in excretory and osmoregulatory tissues in ureotelic taxa, UTs are also expressed ubiquitously in non-ureotelic taxa, and in tissues without a complete ornithine-urea cycle (OUC). We posit that non-OUC production of urea from arginine by arginase, an important pathway to generate ornithine for synthesis of molecules such as polyamines for highly proliferative tissues (e.g. testis, embryos), and neurotransmitters such as glutamate for neural tissues, is an important evolutionary driving force for the expression of UTs in these taxa and tissues.


[Spontaneous level of sister chromatid exchanges in patients with tuberous sclerosis and Recklinghausen's neurofibromatosis].
It is shown that the average number of sister chromatid exchanges (SCE) per one cell in patients with tuberous sclerosis as well as in those with Recklinghausen's neurofibromatosis do not differ from the control. But the non-parametric methods of analysis have revealed differences in the spontaneous level of SCE is patients with tuberous sclerosis, while no such differences were revealed in patients with Recklinghausen's neurofibromatosis.


Early posterior vitreous detachment is associated with LAMA5 dominant mutation.
Extracellular matrix molecular components, previously linked to multisystem syndromes include collagens, fibrillins and laminins. Recently, we described a novel multisystem syndrome caused by the c.9418G>A p.(V3140M) mutation in the laminin alpha-5 (LAMA5) gene, which affects connective tissues of all organs and apparatus in a three generation family. In the same family, we have also reported a myopic trait, which, however, was linked to the Prolyl 4-hydroxylase subunit alpha-2 (P4HA2) gene. Results of investigation on vitreous changes and their pathogenesis are reported in the present study. Nineteen family individuals underwent complete ophthalmic examination including best-corrected visual acuity (BCVA), fundus examination, fundus photography, intraocular pressure measurement, axial length measurement using ocular biometry, Goldmann visual field examination, standard electroretinogram, SD-OCT. Segregation analysis of LAMA5 and P4HA2 mutations was performed in enrolled members. The vitreous alterations fully segregated with LAMA5 mutation in both young and adult family members. Slight reduction of retinal thickness and peripheral retinal degeneration in only two patients were reported. In this work we showed that PVD is a common trait of LAMA5 multisystem syndrome, therefore occurring as an age-unrelated trait. We hypothesize that the p.(V3140M) mutation results in a reduction of retinal inner limiting membrane (ILM) stability, leading to a derangement in the macromolecular structure of the vitreous gel, and PVD. Further investigations will be necessary to elucidate the role of wild type and mutated LAMA5 in the pathogenesis of PVD.


Hormone-regulated transcriptomes: lessons learned from estrogen signaling pathways in breast cancer cells.
Recent rapid advances in next generation sequencing technologies have expanded our understanding of steroid hormone signaling to a genome-wide level. In this review, we discuss the use of a novel genomic approach, global nuclear run-on coupled with massively parallel sequencing (GRO-seq), to explore new facets of the steroid hormone-regulated transcriptome, especially estrogen responses in breast cancer cells. GRO-seq is a high throughput sequencing method adapted from conventional nuclear run-on methodologies, which is used to obtain a map of the position and orientation of all transcriptionally engaged RNA polymerases across the genome with extremely high spatial resolution. GRO-seq, which is an excellent tool for examining transcriptional responses to extracellular stimuli, has been used to comprehensively assay the effects of estrogen signaling on the transcriptome of ERα-positive MCF-7 human breast cancer cells. These studies have revealed new details about estrogen-dependent transcriptional regulation, including effects on transcription by all three RNA polymerases, complex transcriptional dynamics in response to estrogen signaling, and identification novel, unannotated non-coding RNAs. Collectively, these studies have been useful in discerning the molecular logic of the estrogen-regulated mitogenic response.


The potential of dynamic physiological traits in young tomato plants to predict field-yield performance.
To address the challenge of predicting tomato yields in the field, we used whole-plant functional phenotyping to evaluate water relations under well-irrigated and drought conditions. The genotypes tested are known to exhibit variability in their yields in wet and dry fields. The examined lines included two lines with recessive mutations that affect carotenoid biosynthesis, zeta z2083 and tangerine t3406, both isogenic to the processing tomato variety M82. The two mutant lines were reciprocally grafted onto M82, and multiple physiological characteristics were measured continuously, before, during and after drought treatment in the greenhouse. A comparative analysis of greenhouse and field yields showed that the whole-canopy stomatal conductance (gsc) in the morning and cumulative transpiration (CT) were strongly correlated with field measurements of total yield (TY: r2 = 0.9 and 0.77, respectively) and plant vegetative weight (PW: r2 = 0.6 and 0.94, respectively). Furthermore, the minimum CT during drought and the rate of recovery when irrigation was resumed were both found to predict resilience.