Relationship between time in therapeutic range and comparative treatment effect of rivaroxaban and warfarin: results from the ROCKET AF trial.
Time in therapeutic range (TTR) is a standard quality measure of the use of warfarin. We assessed the relative effects of rivaroxaban versus warfarin at the level of trial center TTR (cTTR) since such analysis preserves randomized comparisons. TTR was calculated using the Rosendaal method, without exclusion of international normalized ratio (INR) values performed during warfarin initiation. Measurements during warfarin interruptions >7 days were excluded. INRs were performed via standardized finger-stick point-of-care devices at least every 4 weeks. The primary efficacy endpoint (stroke or non-central nervous system embolism) was examined by quartiles of cTTR and by cTTR as a continuous function. Centers with the highest cTTRs by quartile had lower-risk patients as reflected by lower CHADS2 scores (P<0.0001) and a lower prevalence of prior stroke or transient ischemic attack (P<0.0001). Sites with higher cTTR were predominantly from North America and Western Europe. The treatment effect of rivaroxaban versus warfarin on the primary endpoint was consistent across a wide range of cTTRs (P value for interaction=0.71). The hazard of major and non-major clinically relevant bleeding increased with cTTR (P for interaction=0.001), however, the estimated reduction by rivaroxaban compared with warfarin in the hazard of intracranial hemorrhage was preserved across a wide range of threshold cTTR values. The treatment effect of rivaroxaban compared with warfarin for the prevention of stroke and systemic embolism is consistent regardless of cTTR.


The effect of Phoenix dactylifera pollen on the expression of NRF2, SOD2, CAT, and GPX4 genes, and sperm parameters of fertile and infertile men: A controlled clinical trial.
Oxidative stress is caused by the imbalance occurring between the creation and clearance of the reactive oxygen species (ROS), which is responsible for 30-40% of male infertility. The positive impact of phoenix dactylifera pollen (Date palm pollen, DPP) on the improvement of sperm parameters has been well documented in animal models. For evaluating the effect(s) of DPP on sperm parameters, ROS levels, expression of antioxidant genes, and activity of antioxidant enzymes of infertile men. In this controlled clinical trial, a total of 60 male case with infertility and 20 normospermic fertile men were recruited. Before and after the treatment with DPP, the case were administered 400 mg/kg of gelatinous capsules daily for 30 consecutive days and semen samples were taken. Quantitative real-time polymerase chain reaction was applied for the evaluation of the mRNA expression levels of Nuclear factor erythroid 2-related factor 2(NRF2), superoxide dismutase (SOD2), glutathione peroxidase 4(GPX4), and catalase (CAT) genes. The mRNA expression levels of NRF2, SOD2, GPX4, and CAT (p < 0.05 for all) and significantly increased after treatment with DPP. The increased expressions of all antioxidant genes and enzymes significantly correlated with improvement in semen parameters including count (p = 0.01), motility (p = 0.05), and morphology (p = 0.01) of sperm. A significant correlation between the alteration of SOD2 gene expression and SOD activity, GPX4 and GPX, and CAT were also observed (p = 0.05). DPP can increase the expressions of NRF2, GPX4, SOD2, and CAT genes and also improve the semen quality in infertile men.


A systematic literature review of clinical trials and therapeutic applications of ibogaine.
Iboga and its primary alkaloids, ibogaine and noribogaine, have been of interest to researchers and practitioners, mainly due to their putative efficacy in treating substance use disorders (SUDs). For many SUDs, still no effective pharmacotherapies exist. Distinct psychoactive and somatic effects of the iboga alkaloids set them apart from classic hallucinogens like LSD, mescaline, and psilocybin. The study team performed this systematic review focusing on clinical data and therapeutic interventions involving ibogaine and noribogaine. The team conducted a search for all publications up to December 7, 2020, using PubMed and Embase following PRISMA guidelines. In total, we identified 743 records. In this review, we consider 24 studies, which included 705 individuals receiving ibogaine or noribogaine. This review includes two randomized, double-blind, controlled clinical trials, one double-blind controlled clinical trial, 17 open-label studies or case series (including observational or retrospective studies), three case reports, and one retrospective survey. The published data suggest that ibogaine is an effective therapeutic intervention within the context of SUDs, reducing withdrawal symptoms and craving. Data also point toward a beneficial impact on depressive and trauma-related psychological symptoms. However, studies have reported severe medical complications and deaths, which seem to be associated with neuro- and cardiotoxic effects of ibogaine. Two of these fatalities were described in the 24 studies included in this review. Treatment of SUDs and persisting comorbidities requires innovative treatment approaches. Rapid-onset therapies such as the application of ibogaine may offer novel treatment opportunities for specific individuals. Rigorous study designs within medical settings are necessary to warrant safe application, monitoring, and, possibly, medical intervention.


Evaluating categorisation and clinical relevance of drug-related problems in medication reviews.
We aimed to evaluate the categorisation and clinical relevance of DRPs identified by community pharmacists, and further, to assess the quality of interventions with the patients and the physicians as documented by the pharmacists. 23 Norwegian community pharmacies. Patients with type 2 diabetes were recruited by 24 community pharmacists who performed structured medication reviews based on the patients' drug profiles and patient interviews. The DRPs identified were subsequently categorised. An evaluation group (EG) retrospectively evaluated the reviews. Clinical/practical relevance of each DRP and quality of community pharmacists' intervention with patients and physician were scored. Average agreement between the EG and the community pharmacists was calculated. Internal agreement in the EG was calculated using a modified version of Fleiss' Kappa coefficient. A total of 73 patients were included (mean age 62 years, 52% female, on average prescribed 8.7 drugs). The pharmacists identified 88 DRPs in 43 of the patients. The most common DRPs were adverse drug reactions (22%) and wrong drug or dose used by patient (14%). Anti-diabetic drugs and lipid modifying drugs were associated with the most DRPs. The EG agreed with detection and categorisation of DRPs in more than 80% of the cases. The clinical/practical relevance of the detected DRPs was scored by the EG to be high or medium in 87% of the cases. The quality of the follow-up with patients and physicians was scored to be good or satisfactory in 93 and 98% of the cases, respectively. Pre-defined categories of DRPs supported by structured forms were reliable and valid tools for identifying DRPs. The evaluation demonstrated that community pharmacists were able to identify DRPs of high to medium clinical/practical relevance, and to perform follow-ups of the DRPs with the patients and the physicians with a good or satisfactory quality.


Microbial inactivation and physicochemical properties of ultrasound processed pomegranate juice.
The effects of ultrasound treatment at various amplitudes (50, 75, and 100%) and times (0, 6, 12, 18, 24, and 30 min) on Escherichia coli ATCC 25922 (a surrogate for E. coli O157:H7) and Saccharomyces cerevisiae ATCC 2366 levels and physicochemical characteristics (monomeric anthocyanins, color values, total phenolics, pH, and soluble solids) were determined in pomegranate juice. More than a 5-log inactivation of E. coli ATCC 25922 and a 1.36-log inactivation of S. cerevisiae ATCC 2366 were achieved after 30 min of ultrasound treatment at 100% amplitude. The log-linear and Weibull models were successfully used to estimate the microbial inactivation as a function of ultrasound treatment time (R(2) > 0.97). No significant changes were observed in total phenolics, pH, and soluble solids of the treated juice (P > 0.05). The ultrasound treatment for up to 30 min resulted in more than 92 and 89% anthocyanin retention at 75 and 100% amplitude, respectively. The redness (a*) of the juice did not change significantly after the ultrasound treatment at amplitudes of 75 and 100% for up to 24 and 12 min, respectively. No significant changes in L* and b* values were observed after ultrasound treatment at all amplitudes and after up to 30 min of treatment for 50 and 75% amplitudes. Small differences in juice color were noted based on total color difference scores.


Reduced neural encoding of utility prediction errors in cocaine addiction.
Influential accounts of addiction posit alterations in adaptive behavior driven by deficient dopaminergic prediction errors (PEs), signaling the discrepancy between actual and expected reward. Dopamine neurons encode these error signals in subjective terms, calibrated by individual risk preferences, as "utility" PEs. It remains unclear, however, whether people with drug addiction have PE deficits or their computational source. Here, using an analogous task to prior single-unit studies with known expectancies, we show that fMRI-measured PEs similarly reflect utility PEs. Relative to control participants, people with chronic cocaine addiction demonstrate reduced utility PEs in the dopaminoceptive ventral striatum, with similar trends in orbitofrontal cortex. Dissecting this PE signal into its subcomponent terms attributed these reductions to weaker striatal responses to received reward/utility, whereas suppression of activity with reward expectation was unchanged. These findings support that addiction may fundamentally disrupt PE signaling and reveal an underappreciated role for perceived reward value in this mechanism.


Early discharge in the thrombolytic era: an analysis of criteria for uncomplicated infarction from the Global Utilization of Streptokinase and t-PA for Occluded Coronary Arteries (GUSTO) trial.
This study sought to readdress the definition of uncomplicated myocardial infarction and to apply clinical criteria for early discharge of such patients in the thrombolytic era. Previous studies proposed early hospital discharge at day 7 to 10 after acute myocardial infarction. The potential for earlier discharge of patients with uncomplicated infarction after thrombolysis remains undemonstrated. We defined "uncomplicated infarction" a priori as the absence of death, reinfarction, ischemia, stroke, shock, heart failure (Killip class > 1), bypass surgery, balloon pumping, emergency catheterization or cardioversion or defibrillation in the first 4 hospital days. We applied this definition to 41,021 patients in the Global Utilization of Streptokinase and t-PA for Occluded Coronary Arteries (GUSTO-I) trial. We examined death at 30 days and 1 year and rates of in-hospital reinfarction, heart failure, recurrent ischemia, shock and stroke in the uncomplicated and complicated groups created by application of our definition. We also assessed lengths of hospital and cardiac care unit stay. Application of our clinical criteria yielded 23,497 (57.3%) patients in the uncomplicated group at day 4 with a very low risk of death and in-hospital complications: 30-day mortality 1%, reinfarction 1.7%, heart failure 2.6%, recurrent ischemia 6.7%, shock 0.4% and stroke 0.2%. One-year mortality was 3.6%. The median hospital stay was 9 days (7, 12 [25th, 75th percentiles, respectively]), and the median cardiac care unit stay 3 days (3, 5). Simple clinical characteristics can identify a very low risk post-myocardial infarction population by hospital day 4. Use of these criteria for early discharge planning could substantially reduce length of stay for patients with uncomplicated acute myocardial infarction.


Human IgM monoclonal antibody 16.88: pharmacokinetics and immunogenicity in colorectal cancer patients.
Twenty colorectal cancer patients were given an intravenous injection of human IgM monoclonal antibody (MAb) 16.88 (8 mg) conjugated to 131I for tumor localization. After a 2-week interval, a second injection with 200, 500, or 1000 mg of unlabeled antibody added was given to groups of five patients each. at the end of the 2-hour infusion, 66% of the radioactivity remained in the circulation. Blood clearance of the 131I-labeled MAb 16.88 was biphasic with a mean half-life (T1/2 alpha) of 12 hours and T1/2 beta of 45 hours. Clearance rate was 0.09 L/hour. More than 90% of the 131I in serum was protein bound, with an immunoreactive fraction of 80% in the first 48 hours. Size exclusion chromatography indicated no degradation products other than 131I in serum and urine. The urinary excretion rate of 131I increased to 1.5% of the dose per hour at 24 hours, with 50% of the dose excreted in 34 hours. The pharmacokinetic profile of 131I-labeled MAb 16.88 was neither influenced by the total protein dose of antibody administered nor affected by specific uptake in tumor tissue in individual patients, as determined on early immunoscintigrams. The larger antibody doses showed a slightly slower excretion of 131I. The assays applied to determine immunogenicity were enzyme-linked immunosorbent assay, radioimmunoassay, and the dot-blot assay. They had sensitivities ranging from 5 ng/mL to 0.5 micrograms/mL for goat or rabbit antihuman IgM. The assays did not reveal antihuman antibody responses.


A Bioactive Substance Derived from Brown Seaweeds: Phlorotannins.
Phlorotannins are a type of natural active substance extracted from brown algae, which belong to a type of important plant polyphenol. Phloroglucinol is the basic unit in its structure. Phlorotannins have a wide range of biological activities, such as antioxidant, antibacterial, antiviral, anti-tumor, anti-hypertensive, hypoglycemic, whitening, anti-allergic and anti-inflammatory, etc. Phlorotannins are mainly used in the fields of medicine, food and cosmetics. This paper reviews the research progress of extraction, separation technology and biological activity of phlorotannins, which will help the scientific community investigate the greater biological significance of phlorotannins.


Clinical Characteristics of Patients with Gastroesophageal Reflux Disease Refractory to Proton Pump Inhibitors and the Effects of Switching to 20 mg Esomeprazole on Reflux Symptoms and Quality of Life.
BACKGROUND Refractory gastroesophageal reflux disease (GERD) may deteriorate patient quality of life (QOL) despite proton pump inhibitor (PPI) therapy. MATERIAL AND METHODS Nineteen Japanese institutions were surveyed to determine the clinical characteristics and QOL of patients with refractory GERD. Those patients treated with a conventional PPI were switched to 20 mg esomeprazole for 4 weeks. Symptoms and QOL were assessed using Global Overall Symptom and Gastrointestinal Symptom Rating Scale (GSRS) questionnaires at baseline and at 2 and/or 4 weeks of esomeprazole treatment. RESULTS Of 120 patients who completed the survey, 58 (48.3%) had refractory GERD. Of these, 69.0% were aged ≥ 65 years, 79.3% were prescribed a PPI at a standard or high dose, and 22.4% were prescribed a PPI together with another drug. After switching to esomeprazole, patients reported significant improvements in heartburn, acid regurgitation, and excessive belching at 2 weeks using a symptom diary, as well as the total score, reflux, abdominal pain, and indigestion, which were assessed using the GSRS at 4 weeks. CONCLUSIONS About half of Japanese patients with GERD may be refractory to conventional PPIs. Their reflux-related symptoms are often severe and may impair QOL. Switching to esomeprazole could be used to improve their symptoms and QOL.


Production and characterization of polyclonal anti-S 20499 antibodies: influence of the hapten structure on stereospecificity.
Immunoassays were studied as an alternative to HPLC methods for the stereoselective determination of a chiral drug, S 20499, a new anxiolytic compound that is chemically related to buspirone. The production of highly stereospecific polyclonal antibodies was sought following the construction of appropriately optimized hapten-protein conjugates. This process involved the selection of the structure and the length of the spacer arm used to couple S 20499 to the carrier protein as well as deciding on the location of the coupling site with respect to the chiral center. Two haptens were prepared: one a derivative resembling the original structure of S 20499, with the effective addition of a carboxylic acid group, and a second with the effective addition of a butanoic acid moiety that is supposed to favor stereorecognition. Six stereospecific polyclonal antisera were obtained in rabbits with two groups of antibody families defined in terms of specificity. Both approaches gave high levels of stereospecificity (cross-reactivity towards the optical antipode of S 20499 ranged from 4.1% to < 0.1%). Although it did not decrease the mean apparent affinity constant, the longer spacer improved antibody specificity by decreasing cross-reactions towards dealkylated S 20499 derivatives. Hence, the addition of a four carbon atom bridge should be a valuable tool for increasing antibody stereospecificity with no drawbacks in terms of specificity and affinity. It was also shown that long immunization periods appear to have no effect on the stereospecificity of the antibodies obtained.


Resonant Inelastic X-ray Scattering Amplitudes and Cross Sections in the Algebraic Diagrammatic Construction/Intermediate State Representation (ADC/ISR) Approach.
A scheme has been derived and implemented to gain computational access to the full electronic part of the Kramers-Heisenberg-Dirac (KHD) expression for resonant and nonresonant inelastic scattering amplitudes. Our implementation of this scheme is based on the complex polarization propagator in the algebraic diagrammatic construction (ADC) framework and within its intermediate state representation (ISR). The hierarchy of the second- and third-order ADC/ISR computational schemes known as ADC(2), ADC(2)-x, and ADC(3/2) is considered, and the calculated resonant inelastic X-ray scattering (RIXS) amplitudes and transition strengths for water are in excellent agreement with recent experimental data.


[Chemoluminescence of blood and urine in diffuse appendicular peritonitis in children].
The results of the study of hemiluminescence of native blood, plasma and urine in 56 children with diffuse purulent peritonitis demonstrate that changes of light-sum of the luminescence of the investigated media could reflect aggravation of endogenous intoxication connected with development of organic insufficiency. The usage of blood and urine hemiluminescence for definition of the gravity of endogenous intoxication in diffuse forms of appendicular peritonitis in children is suggested.


Organ geometry channels reproductive cell fate in the Arabidopsis ovule primordium.
In multicellular organisms, sexual reproduction requires the separation of the germline from the soma. In flowering plants, the female germline precursor differentiates as a single spore mother cell (SMC) as the ovule primordium forms. Here, we explored how organ growth contributes to SMC differentiation. We generated 92 annotated 3D images at cellular resolution in Arabidopsis. We identified the spatio-temporal pattern of cell division that acts in a domain-specific manner as the primordium forms. Tissue growth models uncovered plausible morphogenetic principles involving a spatially confined growth signal, differential mechanical properties, and cell growth anisotropy. Our analysis revealed that SMC characteristics first arise in more than one cell but SMC fate becomes progressively restricted to a single cell during organ growth. Altered primordium geometry coincided with a delay in the fate restriction process in katanin mutants. Altogether, our study suggests that tissue geometry channels reproductive cell fate in the Arabidopsis ovule primordium.


Modelling drug adsorption in metal-organic frameworks: the role of solvent.
Solvent plays a key role in biological functions, catalysis, and drug delivery. Metal-organic frameworks (MOFs) due to their tunable functionalities, porosities and surface areas have been recently used as drug delivery vehicles. To investigate the effect of solvent on drug adsorption in MOFs, we have performed integrated computational and experimental studies in selected biocompatible MOFs, specifically, UiO-AZB, HKUST-1 (or CuBTC) and NH2-MIL-53(Al). The adsorption of three drugs, namely, 5-fluorouracil (5-FU), ibuprofen (IBU), and hydroxyurea (HU) were performed in the presence and absence of the ethanol. Our computational predictions, at 1 atmospheric pressure, showed a reasonable agreement with experimental studies performed in the presence of ethanol. We find that in the presence of ethanol the drug molecules were adsorbed at the interface of solvent and MOFs. Moreover, the computationally calculated adsorption isotherms suggested that the drug adsorption was driven by electrostatic interactions at lower pressures (<10-4 Pa). Our computational predictions in the absence of ethanol were higher compared to those in the presence of ethanol. The MOF-adsorbate interaction (U HA) energy decreased with decrease in the size of a drug molecule in all three MOFs at all simulated pressures. At high pressure the interaction energy increases with increase in the MOFs pore size as the number of molecules adsorbed increases. Thus, our research shows the important role played by solvent in drug adsorption and suggests that it is critical to consider solvent while performing computational studies.


Translating words into actions in working memory: The role of spatial-motoric coding.
Research from a working memory perspective on the encoding and temporary maintenance of sequential instructions has established a consistent advantage for enacted over verbal recall. This is thought to reflect action planning for anticipated movements at the response phase. We describe five experiments investigating this, comparing verbal and enacted recall of a series of action-object pairings under different potentially disruptive concurrent task conditions, all requiring repetitive movements. A general advantage for enacted recall was observed across experiments, together with a tendency for concurrent action to impair sequence memory performance. The enacted recall advantage was reduced by concurrent action for both fine and gross concurrent movement with the degree of disruption influenced by both the complexity and the familiarity of the movement. The results are discussed in terms of an output buffer store of limited capacity capable of holding motoric plans for anticipated action.


Identifying barriers to healthcare to reduce health disparity in Zuni Indians using focus group conducted by community health workers.
The Zuni Pueblo is home to an economically disadvantaged population, which faces a public health challenge from the interrelated epidemics of obesity, diabetes and kidney disease. Efforts to decrease the impact of these epidemics have been complicated by historical, economic and cultural barriers, which may limit healthcare utilization. The NIH supported Zuni Health Initiative (ZHI) conducted a study to identify barriers to healthcare in the Zuni Pueblo. Community health representatives (CHRs) led 14 one-hour focus group sessions at which a total of 112 people participated posed unique questions that took into account the Zuni culture to elicit information on perceived barriers to healthcare. Audiotapes were translated and transcribed by bilingual ZHI staff. We reduced the text to thematic categories, constructed a coding dictionary and inserted the text into NVivo 9 program. We identified nine themes emerged regarding the barriers experienced in receiving healthcare and adhering to medical advice. These included distance; transportation; embarrassment; relating to healthcare professionals; navigating the medical system; awareness of available resources; waiting times; adhering to medication; and incentives in health promotion. In conclusion the implementation of culturally appropriate community-based health promotion programs and preventive screening techniques will improve access to healthcare and diminish health disparities.


[Hyperthermic-antiblastic isolation perfusion for advanced melanoma of the limbs. The technic, immediate results and a review of the literature].
Hyperthermic antiblastic isolated perfusion is a method largely used for the treatment of locally advanced limb melanoma. The method requires vascular isolation and hyperthermic perfusion of the limb using an extracorporeal circuit and administering the melphalan as antiblastic drug. Twenty-six patients with primary or recurrent melanoma of the limbs have undergone this treatment at our Institute. There were no cases of operative mortality and systemic toxicity was negligible. The local complications were transitory and no patient showed symptoms of nervous toxicity or permanent functional damage. Two cases of deep thrombophlebitis and two of lymphocele were documented a few months after treatment. Four clinically complete responses, 3 partial and 2 cases of stable disease were observed in the 9 patients treated with unexcised lesions. Our data like the totality of the present experience points to the safety of this method in the therapy of locally advanced limb melanoma. Nevertheless further controlled studies are required to define its role in order to improve survival.


Comparative Genomics of Listeria Sensu Lato: Genus-Wide Differences in Evolutionary Dynamics and the Progressive Gain of Complex, Potentially Pathogenicity-Related Traits through Lateral Gene Transfer.
Historically, genome-wide and molecular characterization of the genus Listeria has concentrated on the important human pathogen Listeria monocytogenes and a small number of closely related species, together termed Listeria sensu strictu. More recently, a number of genome sequences for more basal, and nonpathogenic, members of the Listeria genus have become available, facilitating a wider perspective on the evolution of pathogenicity and genome level evolutionary dynamics within the entire genus (termed Listeria sensu lato). Here, we have sequenced the genomes of additional Listeria fleischmannii and Listeria newyorkensis isolates and explored the dynamics of genome evolution in Listeria sensu lato. Our analyses suggest that acquisition of genetic material through gene duplication and divergence as well as through lateral gene transfer (mostly from outside Listeria) is widespread throughout the genus. Novel genetic material is apparently subject to rapid turnover. Multiple lines of evidence point to significant differences in evolutionary dynamics between the most basal Listeria subclade and all other congeners, including both sensu strictu and other sensu lato isolates. Strikingly, these differences are likely attributable to stochastic, population-level processes and contribute to observed variation in genome size across the genus. Notably, our analyses indicate that the common ancestor of Listeria sensu lato lacked flagella, which were acquired by lateral gene transfer by a common ancestor of Listeria grayi and Listeria sensu strictu, whereas a recently functionally characterized pathogenicity island, responsible for the capacity to produce cobalamin and utilize ethanolamine/propane-2-diol, was acquired in an ancestor of Listeria sensu strictu.


A comparison of sampling methods for examining the laryngeal microbiome.
Shifts in healthy human microbial communities have now been linked to disease in numerous body sites. Noninvasive swabbing remains the sampling technique of choice in most locations; however, it is not well known if this method samples the entire community, or only those members that are easily removed from the surface. We sought to compare the communities found via swabbing and biopsied tissue in true vocal folds, a location that is difficult to sample without causing potential damage and impairment to tissue function. A secondary aim of this study was to determine if swab sampling of the false vocal folds could be used as proxy for true vocal folds. True and false vocal fold mucosal samples (swabbed and biopsied) were collected from six pigs and used for 454 pyrosequencing of the V3-V5 region of the 16S rRNA gene. Most of the alpha and beta measures of diversity were found to be significantly similar between swabbed and biopsied tissue samples. Similarly, the communities found in true and false vocal folds did not differ considerably. These results suggest that samples taken via swabs are sufficient to assess the community, and that samples taken from the false vocal folds may be used as proxies for the true vocal folds. Assessment of these techniques opens an avenue to less traumatic means to explore the role microbes play in the development of diseases of the vocal folds, and perhaps the rest of the respiratory tract.


The hemidesmosomal plaque. I. Characterization of a major constituent protein as a differentiation marker for certain forms of epithelia.
To examine whether constituent proteins of hemidesmosomal structures can be used as markers for certain pathways of epithelial differentiation we have examined the occurrence of the major M- approximately 230,000 plaque protein, the "bullous pemphigoid" (BP) antigen. Several bovine, rat and human tissues and bovine cell culture lines were examined, using different human autoantibody preparations in immunocytochemistry and immunoblotting. We report that this protein, also unequivocally identified by cDNA cloning from expression libraries and DNA sequencing, occurs not only in different stratified epithelia but also, apparently always in hemidesmosomal structures, in urothelium of bladder and the complex epithelia of trachea, bronchus and several glands, notably myoepithelium-containing skin glands, the mammary gland and salivary glands. The protein is absent, however, in all single-layered epithelia and in several tissues reported to have subplasmalemmal densities structurally similar to hemidesmosomes, such as Purkinje fibers of heart, meninges and perineuria. A mammary-gland-derived epithelial cell line (BMGE + H) is particularly rich in hemidesmosomes. This has been used to study the endocytotic uptake of hemidesmosome-containing plasma membrane domains into cytoplasmic vesicles upon detachment of cell sheets during treatment with dispase, a proteolytic enzyme. We propose to use the Mr- approximately 230,000 plaque protein as a marker selective for certain subsets of epithelial cell types and epithelium-derived tumors in studies of fetal and tumor development, including differentiation diagnosis of carcinomas.


Alveolar Hemorrhage in Vasculitis (Primary and Secondary).
Defined by the accumulation of red blood cells into the alveolar space, diffuse alveolar hemorrhage (DAH) is a severe and potentially fatal medical condition requiring careful attention. In contrast to simple extravasation of erythrocytes facilitated by impaired hemostasis or hemodynamic causes, DAH in vasculitis is due to capillaritis, that is, inflammation of capillaries. Dyspnea, hemoptysis, chest infiltrates, and abrupt fall of blood hemoglobin level represent the cardinal features of DAH; yet, hemoptysis is lacking in one-third of cases. Bronchoalveolar lavage, retrieving bright red fluid, is the best diagnostic clue, also excluding infection and other causes of hemoptysis. Although not recommended, lung biopsy is the gold standard for the diagnosis of DAH and pulmonary capillaritis. Pulmonary capillaritis may be primary as in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis or secondary to drugs (especially antithyroid drugs such as propylthiouracil), infections, connective tissue diseases especially systemic lupus erythematosus, or other small vessel vasculitides. Newer toxic causes of drugs of abuse may be difficult to diagnose. Granulomatosis with polyangiitis and microscopic polyangiitis are the most common causes of capillaritis and DAH, whereas DAH is extremely rare in eosinophilic granulomatosis with polyangiitis. When pulmonary capillaritis is not secondary to underlying systemic vasculitis, idiopathic pauci-immune pulmonary capillaritis may be considered, with or without ANCA. Supportive treatment strategy is mandatory in all cases of DAH. Mechanical ventilation and extracorporeal membrane oxygenation may be used in severe cases. Early identification and removal of the putative drug is crucial in drug-induced vasculitis/DAH and may obviate the need for immunosuppressive therapy. High-dose corticosteroids, intravenous cyclophosphamide, and recently rituximab are the mainstay of treatment in vasculitis. Plasma exchange is recommended in anti-glomerular basement membrane disease and in severe DAH associated with systemic lupus erythematosus and is used in selected cases in ANCA-associated vasculitis.


VoxelTrack: Multi-Person 3D Human Pose Estimation and Tracking in the Wild.
We present VoxelTrack for multi-person 3D pose estimation and tracking from a few cameras which are separated by wide baselines. It employs a multi-branch network to jointly estimate 3D poses and re-identification (Re-ID) features for all people in the environment. In contrast to previous efforts which require to establish cross-view correspondence based on noisy 2D pose estimates, it directly estimates and tracks 3D poses from a 3D voxel-based representation constructed from multi-view images. We first discretize the 3D space by regular voxels and compute a feature vector for each voxel by averaging the body joint heatmaps that are inversely projected from all views. We estimate 3D poses from the voxel representation by predicting whether each voxel contains a particular body joint. Similarly, a Re-ID feature is computed for each voxel which is used to track the estimated 3D poses over time. The main advantage of the approach is that it avoids making any hard decisions based on individual images. The approach can robustly estimate and track 3D poses even when people are severely occluded in some cameras. It outperforms the state-of-the-art methods by a large margin on four public datasets including Shelf, Campus, Human3.6 M and CMU Panoptic.


Protein conformation in solution: cross-linking of lysozyme.
The cross-linking of lysozyme by reaction with phenol-2,4-disulfonyl chloride has been effected. The cross-linked protein retained enzymatic activity, has approximately the same molecular weight as native lysozyme, and has essentially the same conformation as native lysozyme as judged by optical rotatory dispersion analysis. The positions of sulfonylation were assigned by a standard degradation sequence; the presence of sulfonamide bonds was confirmed by infrared spectroscopy. Cross-links may thus be introduced without incurring major structural changes in the protein, and certain intramolecular distances that are allowed in the active enzyme may be deduced.


Fluorescence lifetime microscopy of the sodium indicator sodium-binding benzofuran isophthalate in HeLa cells.
The behavior of the sodium indicator sodium-binding benzofuran isophthalate (SBFI) is investigated in HeLa cells by time-resolved fluorescence microscopy. The fluorescence relaxation of SBFI in HeLa cells can be described by a triexponential for intracellular sodium concentration ([Na(+)](i)) between 0 and 90 mM. Changes in [Na(+)](i) affect neither the fluorescence relaxation times (0.21, 0. 60, and 2.7 ns) nor the average decay time (2.2 ns). The preexponential factor of the shortest decay time is negative. However, the ratio of the fluorescence excitation signal at 340 nm to that at 380 nm increases with [Na(+)](i). To elucidate the behavior of SBFI in cells, experiments are performed on SBFI in buffer at various concentrations of sodium, potassium, and bovine serum albumin (BSA) and at various viscosities. The fluorescence decay is triexponential only in the presence of BSA. The relaxation times are independent of [Na(+)] and [BSA]. The preexponential factor of the shortest decay time is negative from a certain [BSA] on, which depends on [Na(+)]. The data indicate that interactions with intracellular components rather than microviscosity influence the SBFI behavior in cells. A model is suggested in which the fluorescence intensities are mainly determined by the signals from the Na(+) subsetSBFI and SBFI subsetprotein complexes.


Resolving fetal hydrops - A rare entity.
Non-immune hydrops fetalis (NIHF) is abnormal accumulation of serous fluid in ≥2 interstitial spaces with no evidence of maternal red cell alloimmunization. Leaving a few treatable conditions, it is generally considered as a sign of poor fetal outcome. Bi-allelic variants in THSD1 have been found to be to be associated with phenotypes ranging from lethal NIHF to persistent edema. Here, we report a family with non-immune hydrops in two successive pregnancies. Whole exome sequencing in second pregnancy identified a homozygous truncating variant in THSD1 (NM_018676:c.892G>T:p.Glu298Ter). Postnatal follow up showed gradual resolution of the accumulated fluid and normal development. This report further strengthens the association of variants in THSD1 with NIHF.


Disorders of the aldosterone synthase and steroid 11beta-hydroxylase deficiencies.
The most potent corticosteroids are 11beta-hydroxylated compounds. In humans, two cytochrome P450 isoenzymes with 11beta-hydroxylase activity, catalysing the biosynthesis of cortisol and aldosterone, are present in the adrenal cortex. CYP11B1, the gene encoding 11beta-hydroxylase (P450c11), is expressed on high levels in the zona fasciculata and is regulated by ACTH. CYP11B2, the gene encoding aldosterone synthase (P450c11Aldo), is expressed in the zona glomerulosa under primary control of the renin-angiotensin system. Aldosterone synthase has 11beta-hydroxylase activity as well as 18-hydroxylase activity and 18-oxidase activity. The substrate for CYP11B2 is 11-deoxycorticosterone, that of CYP11B1 is 11-deoxycortisol. Mutations in CYP11B1 cause congenital adrenal hyperplasia (CAH) due to 11beta-hydroxylase deficiency. This disorder is characterized by androgen excess and hypertension. Mutations in CYP11B2 cause congenital hypoaldosteronism (aldosterone synthase deficiency) which is characterized by life-threatening salt loss, failure to thrive, hyponatraemia and hyperkalaemia in early infancy. Both disorders have an autosomal recessive inheritance. Classical and nonclassical forms of 11beta-hydroxylase deficiency can be distinguished. Studies in heterozygotes for classical 11beta-hydroxylase deficiency show inconsistent results with no or only mild hormonal abnormalities (elevated plasma levels of 11-deoxycortisol after ACTH stimulation). In infants with congenital hypoaldosteronism, a comparable frequency of 18-hydroxylase deficiency (aldosterone synthase deficiency type I) and of 18-oxidase deficiency (aldosterone synthase deficiency type II) can be found. Molecular genetic studies of the CYP11B1 and CYP11B2 genes in 11beta-hydroxylase deficiency or aldosterone synthase deficiency have led to the identification of several mutations. Transfection experiments showed loss of enzyme activity in vitro. In some of the patients with 18-oxidase deficiency (aldosterone synthase deficiency type II) no mutations in the CYP11B2 gene were identified. Refined methods for steroid determination are the basis for the diagnosis of inborn errors of steroidogenesis. Molecular genetic studies are complementary; on the one hand, they have practical importance for the prenatal diagnosis of virilizing CAH forms and on the other hand, they are of theoretical importance in terms of our understanding of the functioning of cytochrome P450 enzymes. Copyrightz1999S.KargerAG, Basel


NAD+ and NADH in ischemic brain injury.
NAD+ and NADH have been emerging as the common mediators of energy metabolism, mitochondrial functions, calcium homeostasis, aging and cell death. NAD+ and NADH can affect cell death by various mechanisms, such as influencing energy metabolism, mitochondrial permeability transition pores, and apoptosis-inducing factor. Because energy failure, calcium disregulation and cell death are the key components in the tissue damaging cascade initiated by cerebral ischemia, it is likely that NAD+ and NADH play significant roles in ischemic brain damage. Many studies, including the findings that poly(ADP-ribose) polymerase-1 mediates ischemic brain injury and that NAD+ administration can decrease ischemic brain damage, have suggested significant roles of NAD+ and NADH in the debilitating illness. However, there is still distinct insufficiency of the information regarding the roles of NAD+ and NADH in ischemic brain injury. Because increasing evidence has indicated critical functions of NAD+ and NADH in various biological processes, future studies on the roles of NAD+ and NADH in cerebral ischemia may expose essential mechanisms underlying ischemic brain injury and suggest novel therapeutic strategies for the illness.


Physical fitness and sports skills in relation to sports injuries. A four-year prospective investigation of sports injuries among physical education students.
In order to study the relationship between physical fitness/sport-specific skills and sports injuries 136 physical education students were studied during their 4-years of training in a prospective investigation. Physical fitness was measured every year using a battery of fitness tests, and the performance marks of a number of sports scored at the exams of the academy were used as parameters for the sport-specific skills. Sports injuries were recorded every 3 weeks on standard forms. Relative risk ratios were calculated between the tertile groups good, average and poor for all variables of physical fitness and sport-specific skills. Injury-proneness was defined for all and for acute and chronic injuries separately near the median number of injuries sustained. In only 6 out of 126 computed relative risks was a significant difference found. Discriminant analysis revealed an explanation of 16%, 14% and 11% of the variance for respectively all, acute and chronic injuries, at which 5 or 6 variables in varying combination were included. From our findings it may be concluded that physical fitness and sport-specific skills have little impact on sports injuries for the following two main reasons. Firstly, subjects at risk for sports injuries participate per definition in sports activities and have consequently developed their fitness and skills compared to the sedentary population. Thus, the range in physical fitness or sports skills in the population at risk is relatively small (physical education students belong to the 7th-10th decile in fitness test scores within a general college student population) and therefore an effect is hard to show. Secondly, the total number of sports injuries is very small and moreover, it should be distributed over several categories for analysis. The favourable advantages of using physical education students to study intrinsic risk factors (comparable and varied sports program, excellent compliance) appeared to be insufficient to compensate for drawbacks of selection.


Titanium and vanadium complexes as anticancer agents.
A series of complexes containing titanium and vanadium as a metal centers have shown to possess a wide spectrum of antitumor properties. These series belong to the non-platinum metal antitumor agents that appear to offer a different alternative for cancer chemotherapy which do not follow mechanism of action of the platinum complexes. The antitumor activity of both titanocene and vanadocene complexes has been established against various animal and xenografted human tumors. The exact mechanism of action for these compounds has not been determined, the target is unknown and even the exact chemical nature of the formulated solutions is still unknown. It has been proposed that these species interact with DNA, inhibiting the cell cycle. However, the antitumor mechanism of the titanocenes is most likely a complex pathway, probably involving a number of different biological molecules related to the transport and delivery of Ti species into cancer cells, and, after hydrolysis, subsequent interaction with nucleic acids and/or proteins and/or other potential coordinating constituents present in the intracellular environment. The tendency to hydrolyze seems to be one of the hypotheses for the tumor-inhibiting potency of the titanocene dihalides. Vanadium compounds exert preventive effects against chemical carcinogenesis on animals, by modifying, mainly, various xenobiotic enzymes, inhibiting, thus, carcinogen-derived active metabolites. The anticarcinogenic effects of vanadium, in combination to its low toxicity, established also, by its administration in humans, suggest vanadium as a candidate antineoplastic agent against human cancer. New complexes being more potent and less toxic favor this perspective. The use of these species as chemotherapeutic agents remains relatively unexplored and waits for future investigation. Research proceeded during the recent decades, enriched our knowledge on the chemical and biochemical properties, as well as the mechanisms of systemic, cellular and molecular antitumor effects of titanium and vanadium compounds.


Interpersonal ambivalence, perceived relationship adjustment, and conjugal loss.
Ambivalence is widely assumed to prolong grief. To examine this hypothesis, the authors developed a measure of ambivalence based on an algorithmic combination of separate positive and negative evaluations of one's spouse. Preliminary construct validity was evidenced in relation to emotional difficulties and to facial expressions of emotion. Bereaved participants, relative to a nonbereaved comparison sample, recollected their relationships as better adjusted but were more ambivalent. Ambivalence about spouses was generally associated with increased distress and poorer perceived health but did not predict long-term grief outcome once initial outcome was controlled. In contrast, initial grief and distress predicted increased ambivalence and decreased Dyadic Adjustment Scale scores at 14 months postloss, regardless of initial scores on these measures. Limitations and implications of the findings are discussed.


Remission induced by an elemental diet in small bowel Crohn's disease.
Seventeen children with active Crohn's disease of the small intestine were entered into a randomised control trial comparing the efficacy of an elemental diet with that of a high dose steroid regimen. Eight children received an elemental diet (Flexical) through a nasogastric tube for six weeks, followed by reintroduction of food over six weeks during which the Flexical was stopped. Seven children were given intramuscular adrenocorticotrophic hormone followed by oral prednisolone with sulphasalazine. Two children were withdrawn from the trial. The elemental diet was equally effective in inducing an improvement in Lloyd-Still disease activity index, erythrocyte sedimentation rate, C reactive protein and albumin concentrations, and body weight as the high dose steroid regimen. Linear growth, assessed from height velocity over six months, was significantly greater in the children receiving an elemental diet.


Ultrasensitive and selective assay of glutathione species in arsenic trioxide-treated leukemia HL-60 cell line by molecularly imprinted polymer decorated electrochemical sensors.
Herein a pair of molecularly imprinted polymer (MIP) modified electrochemical sensors were reported to detect glutathione (GSH) and glutathione disulfide (GSSG) in arsenic trioxide-treated HL-60 cells. MIP film was in situ synthesized onto electrode surface via electro-polymerization in a facile way. The characteristics of the obtained sensors were investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Both GSH-MIP and GSSG-MIP sensors exhibit the relatively wide linear detection range and low detection limit of 1.33 × 10(-10) M (S/N=3). It is found that N-acetylcysteine and DL-homocysteine, the precursors of GSH, show little influence on the detection of glutathione species, nor did the reactants of arsenite and GSH. Such strategies were successfully applied to discriminate GSH and GSSG in cell samples with acceptable recoveries of 92.0-109.1%, and the results are comparable with classic o-phthalaldehyde fluorospectrophotometry. Moreover, the presented sensors allow for easy disclosure of the reversion of malignant phenotype in leukemia cells via glutathione species analysis.


Self-Perceptions and Factors Associated With Being Put Down at School Among Middle and High School Students.
This study assessed the prevalence of verbally being put down by others and intrapersonal and normative factors associated with being put down by others among middle and high school students. Students (N = 1,027) completed the Adolescent Health Risk Behavior Survey. Over 16% of participants reported being put down at school. Students who identified with the negative words confused, unattractive, dull, and careless and identified less with the positive words popular, smart, considerate, cool, and self-confident reported being put down by others. High school students were less likely to be put down. As students scored higher on the Negative Self-Description Scale, their odds of being put down increased. As students scored higher on the Positive Self-Description Scale, their odds of being put down decreased. Students who perceived their friends drinking alcohol regularly were less likely to be put down. Strategies to enhance self-perceptions to raise self-awareness and form healthy/positive identities are needed.


Reducing Opioid Bias is Necessary (ROBIN): An Educational Program to Reduce Addiction Stigma.
Prescription opioid and heroin use and addiction is a local, national, and worldwide epidemic plaguing over 11 million people. Attitudes toward substance use among the general public were highlighted as an area that needs to be further investigated and addressed. The Reducing Opioid Bias is Necessary (ROBIN) educational project was developed and presented to 21 participants as a one-time, 4-hour pretest and posttest design that included theoretical background, short videos, a documentary film, and writings from individuals and community members affected by opioid use disorder (OUD). Participants were assessed on their familiarity, attitudes, and effectiveness of the ROBIN educational program on their perceptions toward persons with OUD. Quantitative data collection included demographic information, level of familiarity, the Attitudes to Mental Illness Questionnaire, and the Drug and Drug Problems Perceptions Questionnaire. Qualitative measures included reflective writing and focus group discussion 2 months after the program. Quantitative measures were completed using descriptive statistics, and qualitative data were sorted for common themes. Spearman's rank rho of 0.947 (p < .05) indicated less familiarity and more bias attitudes toward people with OUD. The paired-sample t test on the Drug and Drug Problems Perceptions Questionnaire indicated a significant decrease in participant bias in the posttest (M = 59.33, SD = 16.48) when compared with the pretest (M = 74.86, SD = 19.38), t(20) = 5.17, p = .00002. The qualitative data from the written reflection and focus group indicated strong changes in participants' overall understanding, empathy, and desire for promoting the educational program. The ROBIN program can reduce bias among community and healthcare subjects. Limitations to the study should be addressed in future research on this topic.


Yeast recombination factor Rdh54 functionally interacts with the Rad51 recombinase and catalyzes Rad51 removal from DNA.
The Saccharomyces cerevisiae RDH54-encoded product, a member of the Swi2/Snf2 protein family, is needed for mitotic and meiotic interhomologue recombination and DNA repair. Previous biochemical studies employing Rdh54 purified from yeast cells have shown DNA-dependent ATP hydrolysis and DNA supercoiling by this protein, indicative of a DNA translocase function. Importantly, Rdh54 physically interacts with the Rad51 recombinase and promotes D-loop formation by the latter. Unfortunately, the low yield of Rdh54 from the yeast expression system has greatly hampered the progress on defining the functional interactions of this Swi2/Snf2-like factor with Rad51. Here we describe an E. coli expression system and purification scheme that together provide milligram quantities of nearly homogeneous Rdh54. Using this material, we demonstrate that Rdh54-mediated DNA supercoiling leads to transient DNA strand opening. Furthermore, at the expense of ATP hydrolysis, Rdh54 removes Rad51 from DNA. We furnish evidence that the Rad51 binding domain resides within the N terminus of Rdh54. Accordingly, N-terminal truncation mutants of Rdh54 that fail to bind Rad51 are also impaired for functional interactions with the latter. Interestingly, the rdh54 K352R mutation that ablates ATPase activity engenders a DNA repair defect even more severe than that seen in the rdh54Delta mutant. These results provide molecular information concerning the role of Rdh54 in homologous recombination and DNA repair, and they also demonstrate the functional significance of Rdh54.Rad51 complex formation. The Rdh54 expression and purification procedures described here should facilitate the functional dissection of this DNA recombination/repair factor.


Formation of the rocky mountains, Western United States: a continuum computer model.
One hypothesis for the information of the Rocky Mountain structures in late Cretaceous through Eocene time is that plate of oceanic lithosphere was underthrust horizontally along the base of the North American lithosphere. The horizontal components of the motion of this plate are known from paleomagnetism, and the edge of the region of flat slab can estimated from reconstructed patterns of volcanism. New techniques of finite-element modeling allow prediction of the thermal and mechanical effects of horizontal subduction on the North American plate. A model that has a realistic temperature-dependent rheology and a simple plane-layered initial condition is used to compute the consequences of horizontal underthrusting in the time interval 75 million to 30 million years before present. Successful prediction of this model include (i) the location, amount, and direction of horizontal shortening that has been inferred from Laramide structures; (ii) massive transport of lower crust from southwest to northeast; (iii) the location and timing of the subsequent extension in metamorphic core complexes and the Rio Grande rift; and (iv) the total area eventually involved in Basin-and-Range style extension. In a broad sense, this model has predicted the belt of Laramide structures, the transport of crust from the coastal region to the continental interior, the subsequent extension in metamorphic core complexes and the Rio Grande rift, and the geographic region of late Tertiary Basin-and-Range extension. Its principal defects are that (i) many events are predicted about 5 million to 10 million years too late and (ii) the wave of crustal thickening does not travel far enough to the east. Reasonable modifications to the oceanic plate kinematics and rheologies that were assumed may correct these defects. The correspondence of model predictions to actual geology is already sufficiently close to show that the hypothesis that horizontal subduction caused the Laramide orogeny is probably correct. The Rocky Mountain thrust and reverse faults formed in an environment of east-west to northeast-southwest compressive stress that was caused by the viscous coupling between the oceanic plate and the base of the North American crust. Nonuniform crustal thickening by simple-shear transport also caused relative uplifts; therefore, this model is consistent with both of the range-forming mechanisms that have been inferred (1). A new proposal that arises from this simulation is that horizontal subduction also caused the subsequent extensional Basin-and-Range taphrogeny by stripping away the mantle lithosphere so that the crust was exposed to hot asthenosphere after the oceanic slab dropped away.


The effect of anxiety on learning outcomes post-CABG.
Post-operative coronary artery bypass graft (CABG) patients require educational interventions to support recovery and prevention of surgical complications. However, the effectiveness of these interventions is questionable, as stress related to the hospitalization process can result in increased levels of anxiety that may impact on the success of the education. The purpose of this study was to examine the relationship between anxiety and the achievement of knowledge, use of self-care behaviours, and management of symptoms. A descriptive correlation design was used, which included a convenience sample. Results indicate statistically significant correlations between anxiety and the outcomes ofinterest. Implications for practice include provision of educational interventions at times when anxiety levels are low.


[Possible mechanism of cannabinoid-mediated modulation of signal transduction through the basal ganglia].
A possible mechanism of cannabinoid-mediated akinesia is suggested. This effect is proposed to be the consequence of a decrease in LTD/LTP in cortical inputs to striatopallidal/striatonigral cells in the matrix due to CB1 receptor activation. In addition, cannabinoids can attenuate locomotor activity due to a reducing of glutamate/GABA release from axon terminals of subthalamic nucleus/striatonigral cells of matrix and subsequent decrease/increase in the activity of neurons of globus pallidus/substantia nigra pars reticulata. Cannabinoid-mediated rise of dopamine release might be a result of a decrease of dopamine neuron inhibition by striatonigral cells of striosomes. It follows from the suggested mechanism that an inactivation (activation) of CB1 receptors leading to rise (lowering) of the motor activity can be useful for treatment of Parkinson (Huntington) disease.


HIV Envelope Trimer Specific Immune Response Is Influenced by Different Adjuvant Formulations and Heterologous Prime-Boost.
The development of a preventive vaccine against human immunodeficiency virus (HIV-1) infection is the most efficient method to control the epidemic. The ultimate goal is to develop a vaccine able to induce specific neutralizing, non-neutralizing antibodies and cellular mediated immunity (CMI). Humoral and CMI responses can be directed to glycoproteins that are normally presented as a trimeric spike on the virus surface (gp140). Despite safer, subunit vaccines are normally less immunogenic/effective and need to be delivered together with an adjuvant. The choice of a suitable adjuvant can induce effective humoral and CMI that utterly lead to full protection against disease. In this report, we established a hierarchy of adjuvant potency on humoral and CMI when admixed with the recombinant HIV gp140 trimer. We show that vaccination with gp140 in the presence of different adjuvants can induce high-affinity antibodies, follicular helper T cells and germinal center B cells. The data show that poly (I:C) is the most potent adjuvant to induce specific CMI responses evidenced by IFN-γ production and CD4+/CD8+ T cell proliferation. Furthermore, we demonstrate that combining some adjuvants like MPL plus Alum and MPL plus MDP exert additive effects that impact on the magnitude and quality of humoral responses while mixing MDP with poly (I:C) or with R848 had no impact on total IgG titers but highly impact IgG subclass. In addition, heterologous DNA prime- protein boost yielded higher IgG titers when compare to DNA alone and improved the quality of humoral response when compare to protein immunization as evidenced by IgG1/IgG2a ratio. The results presented in this paper highlight the importance of selecting the correct adjuvant-antigen combination to potentiate desired cells for optimal stimulation.


Horizontal gene transfer in the innovation and adaptation of land plants.
Horizontal gene transfer (HGT) has been well documented in prokaryotes and unicellular eukaryotes, but its role in plants and animals remains elusive. In a recent study, we showed that at least 57 families of nuclear genes in the moss Physcomitrella patens were acquired from prokaryotes, fungi or viruses and that HGT played a critical role in plant colonization of land. In this paper, we categorize all acquired genes based on their putative functions and biological processes, and further address the importance of HGT in plant innovation and evolution.


Association Between Diseases and Symptoms Diagnosed in Primary Care and the Subsequent Specific Risk of Multiple Sclerosis.
Previous studies have reported a possible prodrome in multiple sclerosis (MS) defined by non-specific symptoms including mood disorder or genito-urinary symptoms and increased health care use detected several years before diagnosis.This study aimed to evaluate agnostically the associations between diseases and symptoms diagnosed in primary care and the risk of multiple sclerosis (MS) relative to controls and two other autoimmune inflammatory diseases with similar population characteristics, namely lupus and Crohn's disease. A case-control study was conducted using electronic health records from the Health Improvement Network database in the UK and France. We agnostically assessed the associations between 113 diseases and symptoms in the five years before and after diagnosis in patients with subsequent diagnosis of MS. Individuals with a diagnosis of MS were compared to individuals without MS, and individuals with two other auto-immune diseases, Crohn's disease and lupus. The study population consisted of patients with MS (n= 20,174), patients without MS (n=54,790), patients with Crohn's disease (n=30,477) or patients with lupus (n=7,337). Twelve ICD-10 codes were significantly positively associated with the risk of MS compared to controls without MS. After considering ICD-10 codes suggestive of neurological symptoms as the first diagnosis of MS, five ICD-10 codes remained significantly associated with MS: depression (UK OR 1.22 [95%CI 1.11-1.34]), sexual dysfunction (1.47 [1.11-1.95]), constipation (1.5 [1.27-1.78]), cystitis (1.21 [1.05-1.39]), and urinary tract infections of unspecified site (1.38 [1.18-1.61]). However, none of these conditions was selectively associated with MS in comparisons with both lupus and Crohn's disease. All five ICD-10 codes identified were still associated with MS during the five years after diagnosis. We identified 5 health conditions associated with subsequent MS diagnosis, which may be considered not only prodromal but also early-stage symptoms. However, these health conditions overlap with prodrome of two other autoimmune diseases, hence they lack specificity to MS.


Virus-inclusive single-cell RNA sequencing reveals the molecular signature of progression to severe dengue.
Dengue virus (DENV) infection can result in severe complications. However, the understanding of the molecular correlates of severity is limited, partly due to difficulties in defining the peripheral blood mononuclear cells (PBMCs) that contain DENV RNA in vivo. Accordingly, there are currently no biomarkers predictive of progression to severe dengue (SD). Bulk transcriptomics data are difficult to interpret because blood consists of multiple cell types that may react differently to infection. Here, we applied virus-inclusive single-cell RNA-seq approach (viscRNA-Seq) to profile transcriptomes of thousands of single PBMCs derived early in the course of disease from six dengue patients and four healthy controls and to characterize distinct leukocyte subtypes that harbor viral RNA (vRNA). Multiple IFN response genes, particularly MX2 in naive B cells and CD163 in CD14+ CD16+ monocytes, were up-regulated in a cell-specific manner before progression to SD. The majority of vRNA-containing cells in the blood of two patients who progressed to SD were naive IgM B cells expressing the CD69 and CXCR4 receptors and various antiviral genes, followed by monocytes. Bystander, non-vRNA-containing B cells also demonstrated immune activation, and IgG1 plasmablasts from two patients exhibited clonal expansions. Lastly, assembly of the DENV genome sequence revealed diversity at unexpected sites. This study presents a multifaceted molecular elucidation of natural dengue infection in humans with implications for any tissue and viral infection and proposes candidate biomarkers for prediction of SD.


Rapid self-assembly of core-shell organosilicon microcapsules within a microfluidic device.
The preparation of hierarchically structured organosilicon microcapsules from commercially available starting materials is described. Using a microfluidic device, an emulsion of dichlorodiphenylsilane is formed in a continuous phase of aqueous glycerol. The silane droplets undergo hydrolysis, condensation, and crystallization within minutes to form self-assembled, core-shell microcapsules. The microparticles have been characterized with light and electron microscopy, nuclear magnetic resonance spectroscopy (NMR), diffusion-ordered NMR spectroscopy (DOSY), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and powder X-ray diffraction (XRD). The characterization data show that the microcapsule walls consist of amorphous, oligomeric poly(diphenylsiloxane) surrounded by a spiny layer of crystalline diphenylsilanediol. Glycerol is occluded within the wall material but is not covalently bound to the silicon components. Glycerol is a crucial element for producing low-dispersity microcapsules with well-ordered surface spines, as the use of methyl cellulose as viscomodifier yields amorphous surfaces.


Protective effects of anti-HMGB1 monoclonal antibody on lung ischemia reperfusion injury in mice.
During ischemia reperfusion (IR) injury, high mobility group box 1 (HMGB1), a chromatin binding protein, is released from necrotic cells and triggers inflammatory responses. We assessed the therapeutic effect of a neutralizing anti-HMGB1 monoclonal antibody (mAb) on lung IR injury. A murine hilar clamp model of IR was used, where mice were divided into sham and IR groups with intravenous administration of anti-HMGB 1 mAb or control mAb. We analyzed the effect of anti-HMGB1 mAb against IR injury by assessing lung oxygenation, lung injury score, neutrophil infiltration, expression of proinflammatory cytokines and chemokines, levels of mitogen-activated protein kinase (MAPK) signaling, and measurement of apoptotic cells. Anti-HMGB1 mAb significantly decreased the plasma level of HMGB1 elevated by IR. The severity of IR injury represented by oxygenation capacity, lung injury score, and neutrophil infiltration was significantly improved by anti-HMGB1 mAb treatment. The expression of proinflammatory factors, including IL-1β, IL-6, IL-12, TNF-α, CXCL-1, and CXCL-2, and phosphorylation of p38 MAPK were both significantly reduced by anti-HMGB1 mAb treatment. Furthermore, anti-HMGB1 mAb treatment suppressed apoptosis, as determined through TUNEL assays. Overall, anti-HMGB1 mAb ameliorated lung IR injury by reducing inflammatory responses and apoptosis. Our findings indicate that anti-HMGB1 mAb has potential for use as a therapeutic to improve IR injury symptoms during lung transplantation.


Effect of fatigue on joint position sense of the knee.
This study was designed to test position sense of the knee joint before and after fatigue in order to determine whether muscle or capsular receptors are the primary sensors for joint position sense. Reproduction of passive positioning and detection of the onset of motion (kinesthesia) were employed to measure joint position sense. Eleven subjects underwent joint position sense measurement before and after a fatigue protocol. A significant worsening of reproduction of knee joint angle after fatigue was noted (p less than 0.05). Threshold (kinesthesia) showed no statistically significant change after fatigue. A significant correlation of reproduction measurements and threshold measurements prior to fatigue (p less than 0.01) demonstrated that the same neural mechanism is applicable in the rested state, but these variables did not correlate significantly after fatigue. There was a significant correlation between reproduction measurements before and after fatigue (p = 0.018), while no correlation was seen for the pre- and postfatigue threshold measurements, suggesting a change in the neural path after fatigue. Since both tests of joint position sense are affected by fatigue, we conclude that muscle receptors are a prominent, if not primary, determinant of joint position sense, and capsular receptors may have a secondary role. Reproduction ability is decreased, presumably through the loss of efficiency of muscle receptors. The threshold data suggest a change in the mechanism of appreciation after fatigue, possibly due to increased sensitivity of capsular receptors from muscle-fatigue-induced laxity.


A Commercial Probiotic Induces Tolerogenic and Reduces Pathogenic Responses in Experimental Autoimmune Encephalomyelitis.
Previous studies in experimental autoimmune encephalomyelitis (EAE) models have shown that some probiotic bacteria beneficially impact the development of this experimental disease. Here, we tested the therapeutic effect of two commercial multispecies probiotics-Lactibiane iki and Vivomixx-on the clinical outcome of established EAE. Lactibiane iki improves EAE clinical outcome in a dose-dependent manner and decreases central nervous system (CNS) demyelination and inflammation. This clinical improvement is related to the inhibition of pro-inflammatory and the stimulation of immunoregulatory mechanisms in the periphery. Moreover, both probiotics modulate the number and phenotype of dendritic cells (DCs). Specifically, Lactibiane iki promotes an immature, tolerogenic phenotype of DCs that can directly induce immune tolerance in the periphery, while Vivomixx decreases the percentage of DCs expressing co-stimulatory molecules. Finally, gut microbiome analysis reveals an altered microbiome composition related to clinical condition and disease progression. This is the first preclinical assay that demonstrates that a commercial probiotic performs a beneficial and dose-dependent effect in EAE mice and one of the few that demonstrates a therapeutic effect once the experimental disease is established. Because this probiotic is already available for clinical trials, further studies are being planned to explore its therapeutic potential in multiple sclerosis patients.


A decade of missile-induced vascular trauma.
During 10 years of serious strife the Royal Victoria Hospital, Belfast, has dealt with trauma inflicted by a wide variety of missiles. Injuries involving 316 vessels were sustained by 192 patients. A group of "knee-cappings' accounted for a large proportion of popliteal vessel injuries. A majority of patients also suffered damage to other regions of the body. Hypotension was a notable feature and several patients arrived in extremis, particularly those with injuries to a major vessel trunks and vital organs. The benefits of rapid evacuation in relation to ischaemia time are discussed. The operative management of a wide range of arterial and venous injuries is described. The use of simple and compound vein grafts allowed adequate excision of damaged vessels. The advantages of meticulous repair, timely fasciotomy, and early reintervention are reflected in the results, contributing to the small lower-lib amputation rate of 5.1%.


Membrane-associated transcription factor peptidase, site-2 protease, antagonizes ABA signaling in Arabidopsis.
Abscisic acid plays important roles in maintaining seed dormancy while gibberellins (GA) and other phytohormones antagonize ABA to promote germination. However, how ABA signaling is desensitized during the transition from dormancy to germination is still poorly understood. We functionally characterized the role of membrane-associated transcription factor peptidase, site-2 protease (S2P), in ABA signaling during seed germination in Arabidopsis. Genetic analysis showed that loss-of-function of S2P conferred high ABA sensitivity during seed germination, and expression of the activated form of membrane-associated transcription factor bZIP17, in which the transmembrane domain and endoplasmic reticulum (ER) lumen-facing C-terminus were deleted, in the S2P mutant rescued its ABA-sensitive phenotype. MYC and green fluorescent protein (GFP)-tagged bZIP17 were processed and translocated from the ER to the nucleus in response to ABA treatment. Furthermore, genes encoding negative regulators of ABA signaling, such as the transcription factor ATHB7 and its target genes HAB1, HAB2, HAI1 and AHG3, were up-regulated in seeds of the wild-type upon ABA treatment; this up-regulation was impaired in seeds of S2P mutants. Our results suggest that S2P desensitizes ABA signaling during seed germination through regulating the activation of the membrane-associated transcription factor bZIP17 and therefore controlling the expression level of genes encoding negative regulators of ABA signaling.


Reconstruction of localized fluorescent target from multi-view continuous-wave surface images of small animal with lp sparsity regularization.
Fluorescence diffuse optical tomography using a multi-view continuous-wave and non-contact measurement system and an algorithm incorporating the lp (0 < p ≤ 1) sparsity regularization reconstructs a localized fluorescent target in a small animal. The measurement system provides a total of 25 fluorescence surface 2D-images of an object, which are acquired by a CCD camera from five different angles of view with excitation from five different angles. Fluorescence surface emissions from five different angles of view are simultaneously imaged on the CCD sensor, thus leading to fast acquisition of the 25 images within three minutes. The distributions of the fluorophore are reconstructed by solving the inverse problem based on the photon diffusion equations. In the reconstruction process incorporating the lp sparsity regularization, the regularization term is reformulated as a differentiable function for gradient-based non-linear optimization. Numerical simulations and phantom experiments show that the use of the lp sparsity regularization improves the localization of the target and quantitativeness of the fluorophore concentration. A mouse experiment demonstrates that a localized fluorescent target in a mouse is successfully reconstructed.


Synthesis, spectroscopic and biological studies on the new symmetric Schiff base derived from 2,6-diformyl-4-methylphenol with N-aminopyrimidine.
A phenol based novel Schiff base polydentate symmetric ligand was prepared. The complexes were prepared by reacting ligand and the metal chlorides of Cu (II), Ni(II), Co(II) and Fe(II) in methanol to get a series of mononuclear complexes. The complexes were characterized by elemental analyses, conductivity measurements, magnetic susceptibility data, IR, UV-Vis, NMR and API-ES mass spectral data. The mononuclear structure of the complexes was confirmed on the basis of elemental analyses, magnetic susceptibility and API-ES mass spectral data. The ligand and all the metal complexes were evaluated for their antimicrobial activity against four Gram-positive bacteria (Staphylococcus aureus ATCC 6538, S. aureus ATCC 25923, Bacillus cereus ATCC 7064 and Micrococcus luteus ATCC 9345), one Gram-negative bacterium (Escherichia coli ATCC 4230), and three yeast (Candida albicans ATCC 14053, Candida krusei ATCC 6258 and Candida parapsilosis ATCC 22019) strains. Therefore, newly synthesized the ligand and two complexes [(Cu(II) and Co(II)] showed good biological activity against all tested bacteria and yeast strains.


Excess Mortality in Treated and Untreated Hyperthyroidism Is Related to Cumulative Periods of Low Serum TSH.
Cumulative time-dependent excess mortality in hyperthyroid patients has been suggested. However, the effect of antithyroid treatment on mortality, especially in subclinical hyperthyroidism, remains unclarified. We investigated the association between hyperthyroidism and mortality in both treated and untreated hyperthyroid individuals. Register-based cohort study of 235,547 individuals who had at least one serum thyroid-stimulating hormone (TSH) measurement in the period 1995 to 2011 (7.3 years median follow-up). Hyperthyroidism was defined as at least two measurements of low serum TSH. Mortality rates for treated and untreated hyperthyroid subjects compared with euthyroid controls were calculated using multivariate Cox regression analyses, controlling for age, sex, and comorbidities. Cumulative periods of decreased serum TSH were analyzed as a time-dependent covariate. Hazard ratio (HR) for mortality was increased in untreated [1.23; 95% confidence interval (CI), 1.12 to 1.37; P < 0.001], but not in treated, hyperthyroid patients. When including cumulative periods of TSH in the Cox regression analyses, HR for mortality per every 6 months of decreased TSH was 1.11 (95% CI, 1.09 to 1.13; P < 0.0001) in untreated hyperthyroid patients (n = 1137) and 1.13 (95% CI, 1.11 to 1.15; P < 0.0001) in treated patients (n = 1656). This corresponds to a 184% and 239% increase in mortality after 5 years of decreased TSH in untreated and treated hyperthyroidism, respectively. Mortality is increased in hyperthyroidism. Cumulative periods of decreased TSH increased mortality in both treated and untreated hyperthyroidism, implying that excess mortality may not be driven by lack of therapy, but rather inability to keep patients euthyroid. Meticulous follow-up during treatment to maintain biochemical euthyroidism may be warranted.


Dually Synergetic Network Hydrogels with Integrated Mechanical Stretchability, Thermal Responsiveness, and Electrical Conductivity for Strain Sensors and Temperature Alertors.
The first example of dually synergetic network hydrogel, which has integrated mechanical stretchability, thermal responsiveness, and electrical conductivity, has been constructed by a versatile and topological co-cross-linking approach. Poly( N-isopropylacrylamide) (PNIPAAm) is introduced as the thermally responsive ingredient, and polyaniline (PANI) is selected as the electrically conductive ingredient. PNIPAAm network is cross-linked by double-bond end-capped Pluronic F127 (F127DA). PANI network is doped and cross-linked by phytic acid. These two ingredients are further mechanically interlocked. Due to the integrated multiple functionalities, the topologically co-cross-linked hydrogels, as will be mentioned as F-PNIPAAm/PANI hydrogels, can be fabricated into resistive-type strain sensors. The strain sensors can achieve a gauge factor of 3.92, a response time of 0.4 s, and a sensing stability for at least 350 cycles and can be further applied for monitoring human motions, including motion of two hands, bending of joints, and even swallowing and pulse rate. Moreover, F-PNIPAAm/PANI hydrogels are utilized to construct efficient temperature alertors based on the disconnection of circuits induced by volume shrinkage at high temperature.


The deubiquitylase USP37 links REST to the control of p27 stability and cell proliferation.
The RE1 silencing transcription factor (REST) is a repressor of neuronal differentiation and its elevated expression in neural cells blocks neuronal differentiation. In this study, we demonstrate a role for REST in the control of proliferation of medulloblastoma cells. REST expression decreased the levels of cyclin-dependent kinase (CDK)NIB/p27, a CDK inhibitor and a brake of cell proliferation in these cells. The reciprocal relationship between REST and p27 was validated in human tumor samples. REST knockdown in medulloblastoma cells derepessed a novel REST target gene encoding the deubiquitylase ubiquitin (Ub)-specific peptidase 37 (USP37). Ectopically expressed wild-type USP37 formed a complex with p27, promoted its deubiquitination and stabilization and blocked cell proliferation. Knockdown of REST and USP37 prevented p27 stabilization and blocked the diminution in proliferative potential that normally accompanied REST loss. Unexpectedly, wild-type USP37 expression also induced the expression of REST-target neuronal differentiation genes even though REST levels were unaffected. In contrast, a mutant of USP37 carrying a site-directed change in a conserved cysteine failed to rescue REST-mediated p27 destabilization, maintenance of cell proliferation and blockade to neuronal differentiation. Consistent with these findings, a significant correlation between USP37 and p27 was observed in patient tumors. Collectively, these findings provide a novel connection between REST and the proteasomal machinery in the control of p27 and cell proliferation in medulloblastoma cells.


Revascularization and functional outcomes after mechanical thrombectomy: an update to key metrics.
The advent of mechanical thrombectomy (MT) has become an effective option for the treatment of acute ischemic stroke in addition to tissue plasminogen activator (tPA). With recent advances in device technology, MT has significantly altered the hospital course and functional outcomes of stroke patients. The authors' goal was to establish the most up-to-date reperfusion and functional outcomes with the evolution of MT technology. The authors conducted a retrospective study of 403 patients who underwent MT for ischemic stroke at their institution from 2010 to 2017. They collected data on patient comorbidities, National Institutes of Health Stroke Scale (NIHSS) score on arrival, tPA administration, revascularization outcomes, and functional outcomes on discharge. In 403 patients, the mean NIHSS score on presentation was 15.8 ± 6.6, with 195 (48.0%) of patients receiving tPA prior to MT. Successful reperfusion (thrombolysis in cerebral infarction score 2B or 3) was achieved in 84.4%. Hemorrhagic conversion with significant mass effect was noted in 9.9% of patients. The median lengths of ICU and hospital stay were 3.0 and 7.0 days, respectively. Functional independence (modified Rankin Scale score 0-2) was noted in 125 (31.0%) patients, while inpatient mortality occurred in 43 (10.7%) patients. As MT has established acute ischemic stroke as a neurosurgical disease, there is a pressing need to understand the hospital course, hospital- and procedure-related complications, and outcomes for this new patient population. The authors provide a detailed account of key metrics for MT with the latest device technology and identify the predictors of unfavorable outcomes and inpatient mortality.


Insurance Transitions and Changes in Physician and Emergency Department Utilization: An Observational Study.
Shopping for health insurance is encouraged as a way to find the most affordable coverage that best meets an enrollee's needs. However, the extent to which individuals switch insurance and subsequent changes in health care utilization that might arise, particularly new physician visits, are not well understood. To examine the relationship between insurance switching and new physician and emergency department visits around the time of a switch. Observational study using a difference-in-differences design to compare those switching insurance carriers with propensity score-matched controls who did not switch, stratified based on whether individuals initially had private or Medicaid insurance coverage. All analyses adjusted for individual and insurance characteristics. Continuously insured, non-elderly individuals with private or Medicaid insurance coverage in Massachusetts from 2010 to 2013. Rates of new primary care and specialist physician visits, as well as rates of emergency department visits. Before matching, among 1,628,057 continuously insured individuals, 418,231 (26%) switched insurance carriers during a 2-year period. Characteristics of switchers and non-switchers were similar after matching (n = 316,343 in each group). After matching, switching plans was associated with a 203% and 47.5% increase in the rate of new primary care physician visits following switching for those initially with Medicaid or private coverage, respectively (both p < 0.001), with a large short-term increase, diminishing over time. Among those with Medicaid coverage, switching was associated with a 14.9% higher rate of ED visits during the month of switching (p < 0.001), but otherwise decreased modestly after switching. Insurance switching is common, and is associated with increased new physician visits and temporarily increased ED use among the publicly insured. As insurance markets become more volatile in the current policy environment, understanding changes in utilization after insurance switching may become increasingly important.


Post-transplant lymphoproliferative disorders.
Post-transplant lymphoproliferative disorder (PTLD) is one of the most dreaded complications of orthotopic transplantation. It consists of a heterogeneous group of lymphoproliferative disorders of varying clonal composition, occurring in immuno-suppressed organ allograft recipients and is frequently due to EBV infection. It is most common in heart/lung transplants followed by heart, liver, and kidney and rarely in bone marrow transplants. Clinically, PTLD can present in a number of ways ranging from features resembling infectious mononucleosis, lymphoproliferative masses involving both nodal and extranodal locations, to a fulminant form characterized by a combination of peripheral lymphadenopathy, severe metabolic acidosis, organ failure or allograft dysfunction. Pathologically PTLD is characterized by a dense inflammatory infiltrate with a spectrum ranging from that found in infectious mononucleosis to a polymorphous B-cell hyperplasia to that of a monomorphous lymphoma. Analysis of EBV is especially useful for the diagnosis of early cases of PTLD. In addition, immunophenotyping to determine the lymphocyte type (B or T cell type) and monoclonality are most helpful in determining the prognosis.


Evaluating the mutagenic activity of targeted endonucleases containing a Sharkey FokI cleavage domain variant in zebrafish.
Synthetic targeted endonucleases such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) have recently emerged as powerful tools for targeted mutagenesis, especially in organisms that are not amenable to embryonic stem cell manipulation. Both ZFNs and TALENs consist of DNA-binding arrays that are fused to the nonspecific FokI nuclease domain. In an effort to improve targeted endonuclease mutagenesis efficiency, we enhanced their catalytic activity using the Sharkey FokI nuclease domain variant. All constructs tested display increased DNA cleavage activity in vitro. We demonstrate that one out of four ZFN arrays containing the Sharkey FokI variant exhibits a dramatic increase in mutagenesis frequency in vivo in zebrafish. The other three ZFNs exhibit no significant alteration of activity in vivo. Conversely, we demonstrate that TALENs containing the Sharkey FokI variant exhibit absent or severely reduced in vivo mutagenic activity in zebrafish. Notably, Sharkey ZFNs and TALENs do not generate increased toxicity-related defects or mortality. Our results present Sharkey ZFNs as an effective alternative to conventional ZFNs, but advise against the use of Sharkey TALENs.


Association of Fusobacterium nucleatum infection with colorectal cancer in Chinese patients.
To investigate Fusobacterium nucleatum (F. nucleatum) abundance in colorectal cancer (CRC) tissues and its association with CRC invasiveness in Chinese patients. The resected cancer and adjacent normal tissues (10 cm beyond cancer margins) from 101 consecutive patients with CRC were collected. Fluorescent quantitative polymerase chain reaction (FQ-PCR) was applied to detect F. nucleatum in CRC and normal tissues. The difference of F. nucleatum abundance between cancer and normal tissues and the relationship of F. nucleatum abundance with clinical variables were evaluated. Fluorescence in situ hybridization (FISH) analysis was performed on 22 CRC tissues with the highest F. nucleatum abundance by FQ-PCR testing to confirm FQ-PCR results. The median abundance of F. nucleatum in CRC tissues [0.242 (0.178-0.276)] was significantly higher than that in normal controls [0.050 (0.023-0.067)] (P < 0.001). F. nucleatum was over-represented in 88/101 (87.1%) CRC samples. The abundance of F. nucleatum determined by 2(-ΔCT) was significantly greater in tumor samples [0.242 (0.178, 0.276)] than in normal controls [0.050 (0.023, 0.067)] (P < 0.001). The frequency of patients with lymph node metastases was higher in the over-abundance group [52/88 (59.1%)] than in the under-abundance group [0/13 (0%)] (P < 0.005). No significant association of F. nucleatum with other clinico-pathological variables was observed (P > 0.05). FISH analysis also found more F. nucleatum in CRC than in normal tissues (median number 6, 25(th) 3, 75(th) 10 vs 2, 25(th) 1, 75(th) 5) (P < 0.01). F. nucleatum was enriched in CRC tissues and associated with CRC development and metastasis.


X-ray diffraction from nonperiodic layered structures with correlations: analytical calculation and experiment on mixed Aurivillius films.
X-ray diffraction from films consisting of layers with different thicknesses, structures and chemical contents is analysed. The disorder is described by probabilities for different sequences of layers. Closed analytical expressions for the diffracted X-ray intensity are obtained when the layers form a stationary Markov chain. The proposed model is applied to the diffraction data from epitaxial sodium bismuth titanate thin films with Aurivillius structure possessing such one-dimensional disorder. In this case, the disorder is caused by a random stacking of three and four perovskite units separated by bismuth oxide interlayers. The results of analytical calculations are in good agreement with the experimental data and indicate that the incorporation of sodium in the Bi(4)Ti(3)O(12) phase causes the formation of a fourth perovskite unit.


Genetic regulation of carotene biosynthesis in selected tomato strains: aspects of beta-carotene biosynthesis and B gene specificity.
A comparative qualitative and quantitative study of the carotene compositions of a high beta-carotene type mutant Priya Darshini-1 (PD-1), high lycopene type, Pusa Ruby (PR) and the F1 hybrid of these two strains was carried out. Increased amounts of beta-zeacarotene and gamma-carotene were realized in the PD-1 parent and in the F1 hybrid. This correlates the increased beta-zeacarotene and gamma-carotene synthesis with increased beta-carotene synthesis. Introduction of gene B (the gene that regulates beta-carotene biosynthesis) to a system lacking it induces considerable amount of beta-carotene synthesis at the expense of lycopene in the hybrid. Based on these observations a 'three point control' for the beta-carotene biosynthesis in tomato fruits is suggested with the gene B capable of promoting two different pathways in the high beta-carotene types. The probable sites of action of the modifier mOB (the gene that regulates expression of B gene) are indicated. Beta-carotene and xanthophyll contents of different parts of tomato plants of a PD-1 type and the PR type F2 segregates of the F1 hybrid (PR X PD-1)F1 have been studied. Results indicate that the action of gene B is highly specific and exclusively oriented towards the fruit and not expressed in any other parts of the tomato plant.


[Morphological criteria of hyperplasia, dysplasia and incipient cancer of the large intestine].
Early stages of adenomas and hyperplastic polyps are observed when the surface of mucous membrane from 76 surgically removed colons was examined under binocular microscope and when biopsies obtained by means of the endomicroscope were studied. A close link between adenomatous and hyperplastic polyps is established and this may indicate the involvement of the latter in carcinogenesis. Besides this, it is shown that epithelial dysplasia in the adenomas may spread to the goblet cells this being followed by the development of peculiar clear cell adenomas which are mainly formed of the goblet cells with signs of altered differentiation. The observation of the adenoma remnants in 38% of the incipient carcinoma cases indicates the close connection between the adenomas and carcinoma.


Spin dynamics in single-molecule magnets and molecular qubits.
Over recent decades, much effort has been made to lengthen spin relaxation/decoherence times of single-molecule magnets and molecular qubits by following different chemical design rules such as maximizing the total spin value, controlling symmetry, enhancing the ligand field or inhibiting key vibrational modes. Simultaneously, electronic structure calculations have been employed to provide an understanding of the processes involved in the spin dynamics of molecular systems and served to refine or introduce new design rules. This review focuses on contemporary theoretical approaches focused on the calculation of spin relaxation/decoherence times, highlighting their main features and scope. Fundamental aspects of experimental techniques for the determination of key Single Molecule Magnet/Spin Qubit properties are also reviewed.


Hip, hip, hooray!
X-rays are best known for showing where bones have fractured, but researchers can also use X-rays to investigate why bones break, which could lead to treatments that reduce the number of elderly people who suffer broken hips.


HSP90 inhibitors potentiate PGF2α-induced IL-6 synthesis via p38 MAP kinase in osteoblasts.
Heat shock protein 90 (HSP90) that is ubiquitously expressed in various tissues, is recognized to be a major molecular chaperone. We have previously reported that prostaglandin F2α (PGF2α), a potent bone remodeling mediator, stimulates the synthesis of interleukin-6 (IL-6) through p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase in osteoblast-like MC3T3-E1 cells, and that Rho-kinase acts at a point upstream of p38 MAP kinase. In the present study, we investigated the involvement of HSP90 in the PGF2α-stimulated IL-6 synthesis and the underlying mechanism in MC3T3-E1 cells. Geldanamycin, an inhibitor of HSP90, significantly amplified both the PGF2α-stimulated IL-6 release and the mRNA expression levels. In addition, other HSP90 inhibitors, 17-allylamino-17demethoxy-geldanamycin (17-AAG) and 17-dimethylamino-ethylamino-17-demethoxy-geldanamycin (17-DMAG) and onalespib, enhanced the PGF2α-stimulated IL-6 release. Geldanamycin, 17-AAG and onalespib markedly strengthened the PGF2α-induced phosphorylation of p38 MAP kinase. Geldanamycin and 17-AAG did not affect the PGF2α-induced phosphorylation of p44/p42 MAP kinase and myosin phosphatase targeting subunit (MYPT-1), a substrate of Rho-kinase, and the protein levels of RhoA and Rho-kinase. In addition, HSP90-siRNA enhanced the PGF2α-induced phosphorylation of p38 MAP kinase. Furthermore, SB203580, an inhibitor of p38 MAP kinase, significantly suppressed the amplification by geldanamycin, 17-AAG or 17-DMAG of the PGF2α-stimulated IL-6 release. Our results strongly suggest that HSP90 negatively regulates the PGF2α-stimulated IL-6 synthesis in osteoblasts, and that the effect of HSP90 is exerted through regulating p38 MAP kinase activation.


Homeopathic treatment for peripheral nerve regeneration: an experimental study in a rat sciatic nerve transection model.
Effects of homeopathic treatment with Hypericum perforatum (Hypericum) on peripheral nerve regeneration was studied using a rat sciatic nerve transection model. Fifty-four male healthy White Wistar rats were divided into three experimental groups (n = 18), randomly: Sham-operation (Sham), control: silicon tube (Sil) and treatment: silicon tube + Hypericum (Sil/Hypericum). In the Sham group after anesthesia left sciatic nerve was exposed through a gluteal muscle incision and after homeostasis muscle was sutured. In the Sil group the left sciatic nerve was exposed the same way and transected proximal to tibio-peroneal bifurcation leaving a 10-mm gap. Proximal and distal stumps were each inserted into a silicone tube. In the Sil/Hypericum group a silicone tube was implanted the same way and each animal received three oral drops of Hypericum 30c twice daily for 1 week. Each group was subdivided into three subgroups of six animals each studied 4, 8, 12 weeks after surgery. Data were analyzed statistically by factorial analysis of variance (ANOVA) and, the Bonferroni test for pair-wise comparisons. Functional study showed faster and better recovery of regenerated axons in Sil/Hypericum than in Sil group (P < 0.05). Gastrocnemius muscle mass in Sil/Hypericum was significantly greater than in Sil group. Morphometric indices of regenerated fibers showed number and diameter of the myelinated fibers in Sil/Hypericum were significantly higher than in control group. Immunohistochemistry, showed the location of reactions to S-100 in Sil/Hypericum was clearly more positive than in Sil group. Hypericum improves functional recovery of peripheral nerve regeneration in rats.


Gender differences in metformin effect on aging, life span and spontaneous tumorigenesis in 129/Sv mice.
Studies in mammals have led to the suggestion that hyperglycemia and hyperinsulinemia are important factors both in aging and in the development of cancer. It is possible that the life-prolonging effects of calorie restriction are due to decreasing IGF-1 levels. A search of pharmacological modulators of insulin/IGF-1 signaling pathway (which mimetic effects of life span extending mutations or calorie restriction) could be a perspective direction in regulation of longevity. Antidiabetic biguanides are most promising among them. The chronic treatment of inbred 129/Sv mice with metformin (100 mg/kg in drinking water) slightly modified the food consumption but failed to influence the dynamics of body weight, decreased by 13.4% the mean life span of male mice and slightly increased the mean life span of female mice (by 4.4%). The treatment with metformin failed influence spontaneous tumor incidence in male 129/Sv mice, decreased by 3.5 times the incidence of malignant neoplasms in female mice while somewhat stimulated formation of benign vascular tumors in the latter.