Interplay between extracellular matrix components and cellular and molecular mechanisms in kidney fibrosis.
Chronic kidney disease (CKD) is characterized by pathological accumulation of extracellular matrix (ECM) proteins in renal structures. Tubulointerstitial fibrosis is observed in glomerular diseases as well as in the regeneration failure of acute kidney injury (AKI). Therefore, finding antifibrotic therapies comprises an intensive research field in Nephrology. Nowadays, ECM is not only considered as a cellular scaffold, but also exerts important cellular functions. In this review, we describe the cellular and molecular mechanisms involved in kidney fibrosis, paying particular attention to ECM components, profibrotic factors and cell-matrix interactions. In response to kidney damage, activation of glomerular and/or tubular cells may induce aberrant phenotypes characterized by overproduction of proinflammatory and profibrotic factors, and thus contribute to CKD progression. Among ECM components, matricellular proteins can regulate cell-ECM interactions, as well as cellular phenotype changes. Regarding kidney fibrosis, one of the most studied matricellular proteins is cellular communication network-2 (CCN2), also called connective tissue growth factor (CTGF), currently considered as a fibrotic marker and a potential therapeutic target. Integrins connect the ECM proteins to the actin cytoskeleton and several downstream signaling pathways that enable cells to respond to external stimuli in a coordinated manner and maintain optimal tissue stiffness. In kidney fibrosis, there is an increase in ECM deposition, lower ECM degradation and ECM proteins cross-linking, leading to an alteration in the tissue mechanical properties and their responses to injurious stimuli. A better understanding of these complex cellular and molecular events could help us to improve the antifibrotic therapies for CKD.


Effects of exepanol-HCl, metoclopramide and domperidone on the lower esophageal sphincter of dogs.
The actions of rac. 3,5-cis-2,3,4,5-tetrahydro-3-methylamino-1-benzoxepine-5-ol hydrochloride (prop. INN exepanol-HCl, KC 2450), metoclopramide and domperidone on the resting pressure of the lower esophageal sphincter (LES) were studied in anesthetized and conscious beagle dogs using pull-through manometrical methods. Exepanol-HCl proved to enhance the LES pressure (LESP) significantly both in anesthetized and conscious dogs. The action of domperidone which was used as reference compound in the anesthetized dog experiments was less pronounced. In conscious dogs the actions of exepanol-HCl and metoclopramide were comparable. Domperidone was not active in this test.


Effects of exposure to environmental chemicals during pregnancy on the development of the male and female reproductive axes.
There is a large body of literature describing effects of environmental chemicals (ECs), many of them anthropogenic with endocrine-disrupting properties, on development in rodent laboratory species, some of which lead to impaired reproduction and adverse health. This literature joins extensive human epidemiological data and opportunistic wildlife findings on health effects of ECs. In contrast, the effect of endocrine disruption on foetal development and reproductive performance in domestic species is less extensively documented. This applies both to domestic farm and to companion species even though the former is critical to food production and the latter share our homes and many aspects of the modern developed human lifestyle. In domestic species, the nature of chemicals exposure in utero and their consequences for animal health and production are poorly understood. A complication in our understanding is that the pace of development, ontogeny and efficiency of foetal and maternal hepatic and placental activity differs between domestic species. In many ways, this reflects the difficulties in understanding human exposure and consequences of that exposure for the foetus and subsequent adult from epidemiological and largely rodent-based data. It is important that domestic species are included in research into endocrine disruption because of their (i) wide variety of exposure to such chemicals, (ii) greater similarity of many developmental processes to the human, (iii) economic importance and (iv) close similarities to developed world human lifestyle in companion species.


1,25(OH)2D3 binding along the rat nephron: autoradiographic study in isolated tubular segments.
1,25-dihydroxyvitamin D3 [1,25(OH)2D3] binding sites were studied along the nephron of rats. The animals were pretreated with the diphosphonate EHDP at doses that inhibit the endogenous production of 1,25(OH)2D3. A dry film autoradiographic technique was applied to tubular segments isolated by microdissection from kidneys incubated in vitro with various concentrations (0.2-12 nM) of [3H]1,25(OH)2D3 in the presence or absence of an excess unlabeled hormone (X200) in order to determine specific binding. Total, nonspecific, and specific labeling were quantified by silver grain counting over cytoplasmic and nuclear areas. Specific nuclear labeling appeared in the cortical ascending limb and papillary collecting tubule at 1 nM. In the distal tubule and, to a lesser extent, in the cortical collecting tubule a specific nuclear labeling was also present, but only at higher concentrations. No specific nuclear labeling was detected in the proximal tubule. All along the nephron, a significant and nonspecific labeling was observed in the cytoplasm, either alone or superimposed over the specific nuclear labeling. In conclusion 1,25(OH)2D3 specific binding sites appear to be localized mainly in the cortical ascending limb of the loop of Henle, in the distal and cortical collecting duct, and in the papillary collecting duct.


Testing for intracycle determinism in pseudoperiodic time series.
A determinism test is proposed based on the well-known method of the surrogate data. Assuming predictability to be a signature of determinism, the proposed method checks for intracycle (e.g., short-term) determinism in the pseudoperiodic time series for which standard methods of surrogate analysis do not apply. The approach presented is composed of two steps. First, the data are preprocessed to reduce the effects of seasonal and trend components. Second, standard tests of surrogate analysis can then be used. The determinism test is applied to simulated and experimental pseudoperiodic time series and the results show the applicability of the proposed test.


Enhanced CO2 methanation at mild temperature on Ni/zeolite from kaolin: effect of metal-support interface.
Catalytic CO2 hydrogenation to CH4 offers a viable route for CO2 conversion into carbon feedstock. The research aimed to enhance CO2 conversion at low temperature and to increase the stability of Ni catalysts using zeolite as a support. NaZSM-5 (MFI), NaA (LTA), NaY (FAU), and NaBEA (BEA) synthesized from kaolin were impregnated with 15% Ni nanoparticles in order to elucidate the effect of surface area, porosity and basicity of the zeolite in increasing Ni activity at mild temperature of ∼200 °C. A highly dispersed Ni catalyst was produced on high surface area NaY meanwhile the mesoporosity of ZSM-5 has no significant effect in improving Ni dispersion. However, the important role of zeolite mesoporosity was observed on the stability of the catalyst. Premature deactivation of Ni/NaA within 10 h was due to the relatively small micropore size that restricted the CO2 diffusion, meanwhile Ni/NaZSM-5 with a large mesopore size exhibited catalytic stability for 40 h of reaction. Zeolite NaY enhanced Ni activity at 200 °C to give 21% conversion with 100% CH4 selectivity. In situ FTIR analysis showed the formation of hydrogen carbonate species and formate intermediates at low temperatures on Ni/NaY, which implied the efficiency of electron transfer from the basic sites of NaY during CO2 reduction. The combination of Ni/NaY interfacial interaction and NaY surface basicity promoted CO2 methanation reaction at low temperature.


Self-calibrated 3D differential phase contrast microscopy with optimized illumination.
3D phase imaging recovers an object's volumetric refractive index from intensity and/or holographic measurements. Partially coherent methods, such as illumination-based differential phase contrast (DPC), are particularly simple to implement in a commercial brightfield microscope. 3D DPC acquires images at multiple focus positions and with different illumination source patterns in order to reconstruct 3D refractive index. Here, we present a practical extension of the 3D DPC method that does not require a precise motion stage for scanning the focus and uses optimized illumination patterns for improved performance. The user scans the focus by hand, using the microscope's focus knob, and the algorithm self-calibrates the axial position to solve for the 3D refractive index of the sample through a computational inverse problem. We further show that the illumination patterns can be optimized by an end-to-end learning procedure. Combining these two, we demonstrate improved 3D DPC with a commercial microscope whose only hardware modification is LED array illumination.


[Non-urothelial neoplasm in patients with infiltrating bladder carcinoma].
To analyze the incidence of non-urothelial neoplasms in patients with infiltrating bladder tumor submitted to radical surgery. A retrospective study was conducted in patients who underwent radical cystoprostatectomy for an infiltrating bladder tumor from 1987-1996. The pathological and follow-up data were analyzed. Of 120 patients submitted to radical cystoprostatectomy for an infiltrating bladder tumor, 23 (19%) had an associated non-urothelial neoplasm: 14 (61%) were metachronous and 9 (39%) were synchronous. Two patients had two secondary tumors (9%). We have found a high incidence of non-urothelial tumor in patients with infiltrating bladder tumor. By order of frequency, these were prostate cancer, gastrointestinal cancer, pulmonary cancer, etc. Studies have ascribed the association of prostate and bladder cancer to the deletion of suppressor genes.


Heritability of delay discounting in adolescence: a longitudinal twin study.
Delay discounting (DD) refers to the preference for smaller immediate rewards over larger but delayed rewards, and is considered to be a distinct component of a broader "impulsivity" construct. Although greater propensity for discounting the value of delayed gratification has been associated with a range of problem behaviors and substance abuse, particularly in adolescents, the origins of individual differences in DD remain unclear. We examined genetic and environmental influences on a real-life behavioral measure of DD using a longitudinal twin design. Adolescent participants were asked to choose between a smaller ($7) reward available immediately and a larger ($10) reward to be received in 7 days. Biometrical genetic analysis using linear structural equation modeling showed significant heritability of DD at ages 12 and 14 (30 and 51%, respectively) and suggested that the same genetic factors influenced the trait at both ages. DD was significantly associated with symptoms of conduct disorder, attention deficit hyperactivity disorder, substance use, and with higher novelty seeking and poor self-regulation. This study provides the first evidence for heritability of DD in humans and suggests that DD can be a promising endophenotype for genetic studies of addiction and externalizing disorders.


Tarsal tunnel syndrome caused by ganglia.
We describe in 30 feet the occurrence of a tarsal tunnel syndrome caused by a ganglion. The presenting symptom was numbness or pain in the toes and the sole with paraesthesiae in the distribution of the medial plantar nerve in 63% of the patients. Swellings which were not palpable were detected by ultrasonography. Twenty-nine patients were treated by operation. Most ganglia originated from the talocalcaneal joint, and five were associated with a talocalcaneal coalition. The surgical outcome was satisfactory in all patients except one who had a further operation for a recurrence of the ganglion.


[Mycobacterium abscessus group lung disease: case reports and review of the literature].
To analyze the clinical manifestations and efficacy of a combination antibiotic therapy including cefoxitin for Mycobacterium abscessus (M.abscessus) group lung disease. We retrospectively analyzed the clinical manifestations of 16 patients with M.abscessus group lung disease, and the responses of 5 cases treated with whole-course clarithromycin and moxifloxacin, initially intensified with intravenous amikacin and cefoxitin therapy for the first 12 weeks. Radiological study showed that 14 patients with M.abscessus group pulmonary disease were classified as nodular bronchiectasis form, and 1 patient as upper lobe cavity form and 1 patient was unclassifiable. The radiological characteristics of M.abscessus group pulmonary disease included multiple micronodules (14/16), bronchiectasis (14/16), tree in bud sign (13/16), cavity (5/16), consolidation (5/16), nodules (5/16), and collapse of lung (3/16). Five cases were treated with a combination antibiotic therapy including cefoxitin. After 3 months treatment for the initial phase, 2 of them got improvement in symptoms, CT manifestations and sputum conversion. Two of them improved in symptoms and CT manifestations, but not in sputum conversion. One case showed no improvement in the initial phase, and continuation therapy also failed to improve symptoms, CT abnormalities or sputum conversion. Nodular bronchiectasis is the main manifestation of M.abscessus group lung disease. The main imaging characteristics included multiple micronodules, bronchiectasis and tree in bud sign. A therapeutic regimen including cefoxitin may be moderately effective in treating M.abscessus group lung disease.


Intermittent Hypoxia Activates Duration-Dependent Protective and Injurious Mechanisms in Mouse Lung Endothelial Cells.
Intermittent hypoxia is a major factor in clinical conditions like the obstructive sleep apnea syndrome or the cyclic recruitment and derecruitment of atelectasis in acute respiratory distress syndrome and positive pressure mechanical ventilation. In vivo investigations of the direct impact of intermittent hypoxia are frequently hampered by multiple co-morbidities of patients. Therefore, cell culture experiments are important model systems to elucidate molecular mechanisms that are involved in the cellular response to alternating oxygen conditions and could represent future targets for tailored therapies. In this study, we focused on mouse lung endothelial cells as a first frontier to encounter altered oxygen due to disturbances in airway or lung function, that play an important role in the development of secondary diseases like vascular disease and pulmonary hypertension. We analyzed key markers for endothelial function including cell adhesion molecules, molecules involved in regulation of fibrinolysis, hemostasis, redox balance, and regulators of gene expression like miRNAs. Results show that short-time exposure to intermittent hypoxia has little impact on vitality and health of cells. At early timepoints and up to 24 h, many endothelial markers are unchanged in their expression and some indicators of injury are even downregulated. However, in the long-term, multiple signaling pathways are activated, that ultimately result in cellular inflammation, oxidative stress, and apoptosis.


Identification and characterization of human organic anion transporter 3 expressing predominantly in the kidney.
A cDNA encoding a multispecific organic anion transporter 3 (hOAT3) was isolated from a human kidney cDNA library. The hOAT3 cDNA consisted of 2179 base pairs that encoded a 543-amino-acid residue protein with 12 putative transmembrane domains. The deduced amino acid sequence of hOAT3 showed 36 to 51% identity to those of other members of the OAT family. Northern blot analysis revealed that hOAT3 mRNA is expressed in the kidney, brain, and skeletal muscle. When expressed in Xenopus laevis oocytes, hOAT3 mediated the transport of estrone sulfate (K(m) = 3.1 microM), p-aminohippurate (K(m) = 87.2 microM), methotrexate (K(m) = 10.9 microM), and cimetidine (K(m) = 57.4 microM) in a sodium-independent manner. hOAT3 also mediated the transport of dehydroepiandrosterone sulfate, ochratoxin A, PGE(2), estradiol glucuronide, taurocholate, glutarate, cAMP and uric acid. Estrone sulfate did not show any trans-stimulatory effects on either influx or efflux of [(3)H]estrone sulfate via hOAT3. hOAT3 interacted with chemically heterogeneous anionic compounds, such as nonsteroidal anti-inflammatory drugs, diuretics, sulfobromophthalein, penicillin G, bile salts and tetraethyl ammonium bromide. The hOAT3 protein was shown to be localized in the basolateral membrane of renal proximal tubules and the hOAT3 gene was determined to be located on the human chromosome 11q12-q13.3 by fluorescent in situ hybridization analysis. These results suggest an important role of hOAT3 in the excretion/detoxification of endogenous and exogenous organic anions in the kidney.


Mapping analysis of the Xylella fastidiosa genome.
A cosmid library was made of the 2.7 Mb genome of the Gram-negative plant pathogenic bacterium Xylella fastidiosa and analysed by hybridisation mapping. Clones taken from the library as well as genomic restriction fragments of rarely cutting enzymes were used as probes. The latter served as a backbone for ordering the initial map contigs and thus facilitated gap closure. Also, the co-linearity of the cosmid map, and thus the eventual sequence, could be confirmed by this process. A subset of the eventual clone coverage was distributed to the Brazilian X.FASTIDIOSA: sequencing network. Data from this effort confirmed more quantitatively initial results from the hybridisation mapping that the redundancy of clone coverage ranged between 0 and 45-fold across the genome, while the average was 15-fold by experimental design. Reasons for this not unexpected fluctuation and the actual gaps are being discussed, as is the use of this effect for functional studies.


Stabilization and structural alteration of the G-quadruplex DNA made from the human telomeric repeat mediated by Tröger's base based novel benzimidazole derivatives.
Ligand-induced stabilization of the G-quadruplex DNA structure derived from the single-stranded 3'-overhang of the telomeric DNA is an attractive strategy for the inhibition of the telomerase activity. The agents that can induce/stabilize a DNA sequence into a G-quadruplex structure are therefore potential anticancer drugs. Herein we present the first report of the interactions of two novel bisbenzimidazoles (TBBz1 and TBBz2) based on Tröger's base skeleton with the G-quadruplex DNA (G4DNA). These Tröger's base molecules stabilize the G4DNA derived from a human telomeric sequence. Evidence of their strong interaction with the G4DNA has been obtained from CD spectroscopy, thermal denaturation, and UV-vis titration studies. These ligands also possess significantly higher affinity toward the G4DNA over the duplex DNA. The above results obtained are in excellent agreement with the biological activity, measured in vitro using a modified TRAP assay. Furthermore, the ligands are selectively more cytotoxic toward the cancerous cells than the corresponding noncancerous cells. Computational studies suggested that the adaptive scaffold might allow these ligands to occupy not only the G-quartet planes but also the grooves of the G4DNA.


Behavior change interventions to improve the health of racial and ethnic minority populations: a tool kit of adaptation approaches.
Adapting behavior change interventions to meet the needs of racial and ethnic minority populations has the potential to enhance their effectiveness in the target populations. But because there is little guidance on how best to undertake these adaptations, work in this field has proceeded without any firm foundations. In this article, we present our Tool Kit of Adaptation Approaches as a framework for policymakers, practitioners, and researchers interested in delivering behavior change interventions to ethnically diverse, underserved populations in the United Kingdom. We undertook a mixed-method program of research on interventions for smoking cessation, increasing physical activity, and promoting healthy eating that had been adapted to improve salience and acceptability for African-, Chinese-, and South Asian-origin minority populations. This program included a systematic review (reported using PRISMA criteria), qualitative interviews, and a realist synthesis of data. We compiled a richly informative data set of 161 publications and twenty-six interviews detailing the adaptation of behavior change interventions and the contexts in which they were undertaken. On the basis of these data, we developed our Tool Kit of Adaptation Approaches, which contains (1) a forty-six-item Typology of Adaptation Approaches; (2) a Pathway to Adaptation, which shows how to use the Typology to create a generic behavior change intervention; and (3) RESET, a decision tool that provides practical guidance on which adaptations to use in different contexts. Our Tool Kit of Adaptation Approaches provides the first evidence-derived suite of materials to support the development, design, implementation, and reporting of health behavior change interventions for minority groups. The Tool Kit now needs prospective, empirical evaluation in a range of intervention and population settings.


A CAV3 microdeletion differentially affects skeletal muscle and myocardium.
Caveolin-3 is the muscle-specific protein product of the caveolin gene family and an integral membrane component of caveolae. Mutations in the gene encoding caveolin-3 (CAV3) underlie four distinct disorders of skeletal muscle: the autosomal dominant form of limb-girdle muscular dystrophy type 1C (LGMD-1C), rippling muscle disease (RMD), sporadic and familial forms of hyperCKemia, and distal myopathy. To characterize a multigenerational Italian family affected by an autosomal dominant myopathic disorder and to assess the expression of caveolin-3, dystrophin, dystrophin-associated glycoproteins, and neuronal nitric oxide synthase in the myocardium of an affected patient. Clinical analysis involved 15 family members. Skeletal muscle expression of sarcolemmal proteins was evaluated by immunohistochemistry and western blot analysis in three affected individuals. Caveolar structures were analyzed through electron microscopy in muscle biopsies and in one heart biopsy. CAV3 genetic analysis showed a heterozygous 3-bp microdeletion (328-330del) in affected individuals, resulting in the loss of a phenylalanine (Phe97del) in the transmembrane domain. In the skeletal muscle, the mutation was associated with severe caveolin-3 deficiency and caveolar disorganization, whereas the expression of the other analyzed muscle proteins was unaltered. Remarkably, caveolin-3 was expressed in myocardium at a level corresponding to about 60% of that of control individuals and was correctly localized at the myocardial cell membranes, with preservation of cardiac myofiber caveolar structures. Clinical analysis revealed the concomitant presence in this family of the following phenotypes: RMD, LGMD, and hyperCKemia. Intrafamilial phenotypic heterogeneity is associated with caveolin-3 Phe97 microdeletion. The molecular network interacting with caveolin-3 in skeletal muscle and heart may differ.


Increased Production of Colanic Acid by an Engineered Escherichia coli Strain, Mediated by Genetic and Environmental Perturbations.
Colanic acid (CA) is a major exopolysaccharide synthesized by Escherichia coli that serves as a constituent of biofilm matrices. CA demonstrates potential applications in the food, cosmetics, and pharmaceutical industry. Moreover, L-fucose, a monomeric constituent of CA, exhibits various physiological activities, such as antitumor, anti-inflammatory, and skin-whitening. Here, the effects of genetic and environmental perturbations were investigated for improving CA production by E. coli. When rcsF, a positive regulator gene of CA synthesis, was expressed in E. coli ΔwaaF, a CA-producing strain constructed previously, the CA titer increased to 3051.2 mg/L as compared to 2052.8 mg/L observed with E. coli ΔwaaF. Among the environmental factors tested, namely, osmotic and oxidative stresses and pH, pH was a primary factor that significantly improved CA production. When the pH of the culture medium of E. coli ΔwaaF + rcsF was maintained at 7, the CA titer significantly increased to 4351.6 mg/L. The CA yield obtained with E. coli ΔwaaF + rcsF grown at pH 7 was 5180.4 mg CA/g dry cell weight, which is the highest yield of CA reported so far. This engineered E. coli system with optimization of environmental conditions can be employed for fast and economically-feasible production of CA.


Religion and beliefs about treating medically unexplained symptoms: a survey of primary care physicians and psychiatrists.
Historical evidence and prior research suggest that psychiatry is biased against religion, and religious physicians are biased against the mental health professions. Here we examine whether religious and non-religious physicians differ in their treatment recommendations for a patient with medically unexplained symptoms. We conducted a national survey of primary care physicians and psychiatrists. We presented a vignette of a patient with medically unexplained symptoms, and experimentally varied whether the patient was religiously observant. We asked whether physicians would recommend six interventions: antidepressant medication, in-office counseling, referral to a psychiatrist, referral to a psychologist or licensed counselor, participation in meaningful relationships and activities, and involvement in religious community. Predictors included the physician's specialty and the physician's attendance at religious services. The response rate was 63% (896 of 1427) primary care physicians and 64% (312 of 487) psychiatrists. We did not find evidence that religious physicians were less likely to recommend mental health resources, nor did we find evidence that psychiatrists were less likely to recommend religious involvement. Primary care physicians (but not psychiatrists) were more likely to recommend that the patient get more involved in their religious community when the patient was more religiously observant, and when the physician more frequently attended services. We did not find evidence that mental health professionals are biased against religion, nor that religious physicians are biased against mental health professionals. Historical tensions are potentially being replaced by collaboration.


Sirolimus reduces vasculopathy but exacerbates proteinuria in association with inhibition of VEGF and VEGFR in a rat kidney model of chronic allograft dysfunction.
Use of the mTOR inhibitor (mTORi) sirolimus to replace calcineurin inhibitors in kidney transplantation has been associated with improved renal function but, in a proportion of cases, also with de novo or exacerbated proteinuria. Experimental deficiency of vascular endothelial growth factor (VEGF) induces proteinuria and mTOR is required for VEGF production and signalling. We therefore explored the impact of sirolimus on the development of chronic allograft dysfunction (CAD) in the rat, with a focus on VEGF biology. Lewis rats received F344 kidney allografts and were treated with 24 weeks of cyclosporine or sirolimus. Controls included allografts treated with cyclosporine for 10 days only and isografts treated with cyclosporine or sirolimus for 24 weeks. Kidney injury (proteinuria and histology) and expression of VEGF and VEGF-receptor (VEGFR; immunohistochemistry, laser capture micro-dissection and quantitative RT-PCR) were assessed. Allograft controls developed proteinuria, tubulointerstitial fibrosis and atrophy, glomerulosclerosis, vasculopathy and leucocyte accumulation. Proteinuria was significantly reduced in both treatment groups but significantly more in cyclosporine treated animals. Tubulointerstitial damage, glomerulosclerosis and leucocyte accumulation were significantly attenuated in both treatment groups; however, vasculopathy was reduced only by sirolimus. Significantly diminished expression of VEGF and VEGFR mRNA and protein was evident in the sirolimus group. In vitro, sirolimus reduced VEGF production by podocytes (P < 0.05) and inhibited VEGF-induced proliferation of podocytes, endothelial and mesangial cells. Cyclosporine and sirolimus retard development of CAD in this rat model. Sirolimus exhibits greater protection against vasculopathy but induces proteinuria; effects are likely to be related to inhibition of VEGF signalling.


Toll-like receptor 4 is involved in inflammatory and joint destructive pathways in collagen-induced arthritis in DBA1J mice.
In rheumatoid arthritis, a significant proportion of cytokine and chemokine synthesis is attributed to innate immune mechanisms. TLR4 is a prominent innate receptor since several endogenous ligands known to activate the innate immune system bind to it and may thereby promote joint inflammation. We generated TLR4 deficient DBA1J mice by backcrossing the TLR4 mutation present in C3H/HeJ strain onto the DBA1J strain and investigated the course of collagen-induced arthritis in TLR4 deficient mice in comparison to wild type littermates. The incidence of collagen- induced arthritis was significantly lower in TLR4 deficient compared to wild type mice (59 percent vs. 100 percent). The severity of arthritis was reduced in the TLR4 deficient mice compared to wild type littermates (mean maximum score 2,54 vs. 6,25). Mice deficient for TLR4 were virtually protected from cartilage destruction, and infiltration of inflammatory cells was reduced compared to wt mice. In parallel to the decreased clinical severity, lower anti-CCP antibody concentrations and lower IL-17 concentrations were found in the TLR4 deficient mice. The study further supports the role of TLR4 in the propagation of joint inflammation and destruction. Moreover, since deficiency in TLR4 led to decreased IL-17 and anti-CCP antibody production, the results indicate a link between TLR4 stimulation and the adaptive autoimmune response. This mechanism might be relevant in human rheumatoid arthritis, possibly in response to activating endogenous ligands in the affected joints.


[Analysis of the pathogenic characteristics of 162 severely burned patients with bloodstream infection].
To analyze the distribution and drug resistance of pathogen isolated from severely burned patients with bloodstream infection, so as to provide reference for the clinical treatment of these patients. Blood samples of 162 severely burned patients (including 120 patients with extremely severe burn) with bloodstream infection admitted into our burn ICU from January 2011 to December 2014 were collected. Pathogens were cultured by fully automatic blood culture system, and API bacteria identification panels were used to identify pathogen. Kirby-Bauer paper disk diffusion method was used to detect the drug resistance of major Gram-negative and -positive bacteria to 37 antibiotics including ampicillin, piperacillin and teicoplanin, etc. (resistance to vancomycin was detected by E test), and drug resistance of fungi to 5 antibiotics including voriconazole and amphotericin B, etc. Modified Hodge test was used to further identify imipenem and meropenem resistant Klebsiella pneumonia. D test was used to detect erythromycin-induced clindamycin resistant Staphylococcus aureus. The pathogen distribution and drug resistance rate were analyzed by WHONET 5.5. Mortality rate and infected pathogens of patients with extremely severe burn and patients with non-extremely severe burn were recorded. Data were processed with Wilcoxon rank sum test. (1) Totally 1 658 blood samples were collected during the four years, and 339 (20.4%) strains of pathogens were isolated. The isolation rate of Gram-negative bacteria, Gram-positive bacteria, and fungi were 68.4% (232/339), 24.5% (83/339), and 7.1% (24/339), respectively. The top three pathogens with isolation rate from high to low were Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa in turn. (2) Except for the low drug resistance rate to polymyxin B and minocycline, drug resistance rate of Acinetobacter baumannii to the other antibiotics were relatively high (81.0%-100.0%). Pseudomonas aeruginosa was sensitive to polymyxin B but highly resistant to other antibiotics (57.7%-100.0%). Enterobacter cloacae was sensitive to imipenem and meropenem, while its drug resistance rates to ciprofloxacin, levofloxacin, cefoperazone/sulbactam, cefepime, piperacillin/tazobactam were 25.0%-49.0%, and those to the other antibiotics were 66.7%-100.0%. Drug resistance rates of Klebsiella pneumoniae to cefoperazone/sulbactam, imipenem, and meropenem were low (5.9%-15.6%, two imipenem- and meropenem-resistant strains were identified by modified Hodge test), while its drug resistance rates to amoxicillin/clavulanic acid, piperacillin/tazobactam, cefepime, cefoxitin, amikacin, levofloxacin were 35.3%-47.1%, and those to the other antibiotics were 50.0%-100.0%. (3) Drug resistance rates of methicillin-resistant Staphylococcus aureus (MRSA) to most of the antibiotics were higher than those of the methicillin-sensitive Staphylococcus aureus (MSSA). MRSA was sensitive to linezolid, vancomycin, and teicoplanin, while its drug resistance rates to compound sulfamethoxazole, clindamycin, minocycline, and erythromycin were 5.3%-31.6%, and those to the other antibiotics were 81.6%-100.0%. Except for totally resistant to penicillin G and tetracycline, MSSA was sensitive to the other antibiotics. Fourteen Staphylococcus aureus strains were resistant to erythromycin-induced clindamycin. Enterococcus was sensitive to vancomycin and teicoplanin, while its drug resistance rates to linezolid, chloramphenicol, nitrofurantoin, and high unit gentamicin were low (10.0%-30.0%), and those to ciprofloxacin, erythromycin, minocycline, and ampicillin were high (60.0%-80.0%). Enterococcus was fully resistant to rifampicin. (4) Fungi was sensitive to amphotericin B, and drug resistance rates of fungi to voriconazole, fluconazole, itraconazole, and ketoconazole were 7.2%-12.5%. (5) The mortality of patients with extremely severe burn was higher than that of patients with non-extremely severe burn. The variety of infected pathogens in patients with extremely severe burn significantly outnumbered that in patients with non-extremely severe burn (Z=-2.985, P=0.005). The variety of pathogen in severely burned patients with bloodstream infection is wide, with the main pathogens as Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa, and the drug resistance situation is grim. The types of infected pathogen in patients with extremely severe burn are more complex, and the mortality of these patients is higher when compared with that of patients with non-extremely severe burn.


Equilibrium gels of low-valence DNA nanostars: a colloidal model for strong glass formers.
Kinetic arrest in colloidal dispersions with isotropic attractive interactions usually occurs through the destabilization of the homogeneous phase and the formation of a non-equilibrium network of jammed particles. Theory and simulations predict that a different route to gelation should become available when the valence of each colloidal particle is suitably reduced. Under these conditions, gelation should be achievable through a reversible sequence of equilibrium states. Here we report the reversible dynamic arrest of a dispersion of DNA-based nanoparticles with anisotropic interactions and a coordination number equal to four. As the temperature is decreased, the relaxation time for density fluctuations slows down by about five orders of magnitude, following an Arrhenius scaling in the entire experimentally accessible temperature window. The system is in thermodynamic equilibrium at all temperatures. Gelation in our system mimics the dynamic arrest of networking atomic strong glass formers such as silica, for which it could thus provide a suitable colloidal model.


Disentangling of Local and Wide-Field Motion Adaptation.
Motion adaptation has been attributed in flying insects a pivotal functional role in spatial vision based on optic flow. Ongoing motion enhances in the visual pathway the representation of spatial discontinuities, which manifest themselves as velocity discontinuities in the retinal optic flow pattern during translational locomotion. There is evidence for different spatial scales of motion adaptation at the different visual processing stages. Motion adaptation is supposed to take place, on the one hand, on a retinotopic basis at the level of local motion detecting neurons and, on the other hand, at the level of wide-field neurons pooling the output of many of these local motion detectors. So far, local and wide-field adaptation could not be analyzed separately, since conventional motion stimuli jointly affect both adaptive processes. Therefore, we designed a novel stimulus paradigm based on two types of motion stimuli that had the same overall strength but differed in that one led to local motion adaptation while the other did not. We recorded intracellularly the activity of a particular wide-field motion-sensitive neuron, the horizontal system equatorial cell (HSE) in blowflies. The experimental data were interpreted based on a computational model of the visual motion pathway, which included the spatially pooling HSE-cell. By comparing the difference between the recorded and modeled HSE-cell responses induced by the two types of motion adaptation, the major characteristics of local and wide-field adaptation could be pinpointed. Wide-field adaptation could be shown to strongly depend on the activation level of the cell and, thus, on the direction of motion. In contrast, the response gain is reduced by local motion adaptation to a similar extent independent of the direction of motion. This direction-independent adaptation differs fundamentally from the well-known adaptive adjustment of response gain according to the prevailing overall stimulus level that is considered essential for an efficient signal representation by neurons with a limited operating range. Direction-independent adaptation is discussed to result from the joint activity of local motion-sensitive neurons of different preferred directions and to lead to a representation of the local motion direction that is independent of the overall direction of global motion.


[Structural and functional characteristics of the sarcoplasmic reticulum membranes of the skeletal muscles in thyrotoxicosis].
A rate of CA2+ accumulation and efficiency of Ca2+-pump were decreased in the fractions of skeletal muscle sarcoplasmic reticulum isolated from rabbits with thyrotoxicosis. Temperature of phase transition in membranes of sarcoplasmic reticulum was changed by 4-5 degrees towards low temperatures. Under conditions of thyrotoxicosis fatty acid composition of total lipids from the sarcoplasmic reticulum fractions was altered as follows: amount of palmitic acid was increased 1.4-fold but oleic and arachidonic acids were decreased 21.8% and 40.5%, respectively. Modification of Ca2+-transport properties in sarcoplasmic reticulum caused by long-term administration of thyroid hormones into animals appears to alter the membrane protein-lipid composition.


An aluminium adjuvant compound with ginseng stem leaf saponins enhances the potency of inactivated Pseudomonas plecoglossicida vaccine in large yellow croaker (Larimichthys crocea).
Large yellow croaker (Larimichthys crocea) farm industry in China suffered from huge economic loss caused by Pseudomonas plecoglossicida infection. Due to multi-antibiotic resistance, efficient vaccines are urgent to be developed to combat this pathogen. In this study, an inactivated vaccine was developed with an aluminium adjuvant (Alum) plus ginseng stem and leaf saponins (GSLS). As a result, the relative percentage survival (RPS) against P. plecoglossicida was up to 67.8 %. Comparatively, RPS of groups that vaccinated with only inactivated vaccine and vaccine containing Alum or Montanide™ 763A as adjuvant were 21.8 %, 32.2 % and 62.1 %, respectively. Assays for total serum protein and serum lysozyme activity in group vaccinated with inactivated vaccine plus Alum + GSLS adjuvant were significantly higher than that in control group. Moreover, specific antibody in serum elicited a rapid and persistent level. According to the expression of some immune related genes, inactivated vaccine plus Alum + GSLS adjuvant induced a stronger cellular immune response which was vital to defend against P. plecoglossicida. In conclusion, our study demonstrated that the compound Alum and GSLS adjuvant is a potential adjuvant system to develop LYC vaccine.


Morphologic demonstration of receptor-mediated endocytosis of epidermal growth factor by isolated bile duct epithelial cells.
It was recently shown that intrahepatic bile duct epithelial cells in situ or after isolation from rat liver have coated pits and vesicles, suggesting that they participate in receptor-mediated endocytosis. Therefore, using a morphologic approach and epidermal growth factor coupled to horseradish peroxidase or colloidal gold as probes, we studied freshly isolated or short-term cultured intrahepatic bile duct epithelial cells prepared from normal rat liver to determine if they participate in receptor-mediated endocytosis. Immunoelectron microscopy using a monoclonal antibody against the epidermal growth factor receptor was also used to examine for the presence of the growth factor receptor on the cells. Immediately after isolation, the cells did not internalize either epidermal growth factor-horseradish peroxidase or epidermal growth factor-colloidal gold; no growth factor receptor could be shown on these cells by immunocytochemistry, either. In contrast, cells cultured for 24 h bound and internalized both epidermal growth factor-horseradish peroxidase and epidermal growth factor-colloidal gold at 37 degrees C and showed growth factor receptors diffusely distributed on the plasma membrane. When cultured cells exposed to epidermal growth factor-colloidal gold were fixed with glutaraldehyde containing saponin and tannic acid, colloidal gold particles were observed in coated pits and in coated and uncoated vesicles. Preincubation of cultured cells with native epidermal growth factor completely blocked the internalization of both epidermal growth factor-horseradish peroxidase and epidermal growth factor-colloidal gold. When rat liver was stained in situ for epidermal growth factor receptor, reaction product was observed by immunoelectron microscopy exclusively on the basal surface of the plasma membrane of the intrahepatic bile duct epithelial cells. These results indicate that bile duct epithelial cells internalize epidermal growth factor by endocytosis via coated pits containing receptors localized in situ exclusively to the basal domain of their plasma membranes. The data demonstrate for the first time that intrahepatic bile duct epithelial cells participate in receptor-mediated endocytosis and raise the possibility that they are a target for epidermal growth factor.


Computational study of the covalent bonding of microcystins to cysteine residues--a reaction involved in the inhibition of the PPP family of protein phosphatases.
Microcystins (MCs) are cyclic peptides, produced by cyanobacteria, that are hepatotoxic to mammals. The toxicity mechanism involves the potent inhibition of protein phosphatases, as the toxins bind the catalytic subunits of five enzymes of the phosphoprotein phosphatase (PPP) family of serine/threonine-specific phosphatases: Ppp1 (aka PP1), Ppp2 (aka PP2A), Ppp4, Ppp5 and Ppp6. The interaction with the proteins includes the formation of a covalent bond with a cysteine residue. Although this reaction seems to be accessory for the inhibition of PPP enzymes, it has been suggested to play an important part in the biological role of MCs and furthermore is involved in their nonenzymatic conjugation to glutathione. In this study, the molecular interaction of microcystins with their targeted PPP catalytic subunits is reviewed, including the relevance of the covalent bond for overall inhibition. The chemical reaction that leads to the formation of the covalent bond was evaluated in silico, both thermodynamically and kinetically, using quantum mechanical-based methods. As a result, it was confirmed to be a Michael-type addition, with simultaneous abstraction of the thiol hydrogen by a water molecule, transfer of hydrogen from the water to the α,β-unsaturated carbonyl group of the microcystin and addition of the sulfur to the β-carbon of the microcystin moiety. The calculated kinetics are in agreement with previous experimental results that had indicated the reaction to occur in a second step after a fast noncovalent interaction that inhibited the enzymes per se.


Evaluation of critical parameters for in vitro skin permeation and penetration studies using animal skin models.
In vitro skin permeation/penetration studies may be affected by many sources of variation. Herein, we aimed to investigate the major critical procedures of in vitro skin delivery studies. These experiments were performed with model drugs according to official guidelines. The influence of skin source on penetration studies was studied as well as the use of a cryopreservation agent on skin freezing evaluated by transepidermal water loss, electrical resistance, permeation/penetration profiles and histological changes of the skin. The best condition for tape stripping procedure was validated through the evaluation of the distribution of corneocytes, mass of stratum corneum (SC) removed and amount of protein removed using finger pressure, a 2kg weight and a roller. The interchangeability of the tape stripping procedures followed by the epidermis and dermis homogenate and the micrometric horizontal cryostat skin sectioning methods were also investigated, besides the effect of different formulations. Noteworthy, different skin sources were able to ensure reliable interchangeability for in vitro permeation studies. Furthermore, an increased penetration was obtained for stored frozen skin compared to fresh skin, even with the addition of a cryoprotectant agent. The best method for tape stripping was the finger pressure followed by the addition of a propylene glycol solvent leading to better SC removal. Finally, no significant difference was found in skin penetration studies performed by different methods suggesting their possible interchangeability.


Factors affecting outcome after incomplete excision of cervical intraepithelial neoplasia.
Conservative treatment for cervical intraepithelial neoplasia (CIN) by ablative or excisional techniques is widely used. However, women with incomplete excision have a variable risk of CIN recurrence. The aim of this study was to identify possible risk factors for recurrence of CIN after large loop excision of the transformation zone (LLETZ) with involved margins of excision. All cases of women treated with LLETZ for CIN between 1989-2000, in whom histological evaluation of the excised specimen revealed extension of CIN to the excision margins, were retrospectively studied. A woman was considered to have recurrence when she had histologically confirmed CIN following a second LLETZ or hysterectomy during the follow-up period. The characteristics that were examined as possible risk factors were age, parity, smoking habit, grade of initial lesion and extension to the endo- or ectocervical margin. Treatment failure was diagnosed in 18 out of 65 (27.7%) women with involved margins. The only characteristic that reached statistical significance was age. The mean age of women with recurrence was 35.8 years, whereas the mean age of women without recurrence was 32.8 years (p = 0.044). Also, a trend was evident in women with high-grade initial lesions (CIN II-III) (p = 0.168) and involvement of the endocervical margins (p = 0.149). No differences were observed between the two groups regarding parity and smoking habit. Increased age is a risk factor for recurrence in women with incomplete excision of CIN after LLETZ. Larger studies are required for definite conclusions.


Identification, nucleotide sequence, and characterization of PspF, the transcriptional activator of the Escherichia coli stress-induced psp operon.
The phage shock protein (psp) operon (pspABCE) of Escherichia coli is strongly induced in response to a variety of stressful conditions or agents such as filamentous phage infection, ethanol treatment, osmotic shock, heat shock, and prolonged incubation in stationary phase. Transcription of the psp operon is driven from a sigma54 promoter and stimulated by integration host factor. We report here the identification of a transcriptional activator gene, designated pspF, which controls expression of the psp operon in E. coli. The pspF gene was identified by random miniTn10-tet transposon mutagenesis. Insertion of the transposon into the pspF gene abolished sigma54-dependent induction of the psp operon. The pspF gene is closely linked to the psp operon and is divergently transcribed from one major and two minor sigma 70 promoters, pspF encodes a 37-kDa protein which belongs to the enhancer-binding protein family of sigma54 transcriptional activators. PspF contains a catalytic domain, which in other sigma54 activators would be the central domain, and a C-terminal DNA-binding domain but entirely lacks an N-terminal regulatory domain and is constitutively active. The insertion mutant pspF::mTn10-tet (pspF877) encodes a truncated protein (PspF delta HTH) that lacks the DNA-binding helix-turn-helix (HTH) motif. Although the central catalytic domain is intact, PspF delta HTH at physiological concentration cannot activate psp expression. In the absence of inducing stimuli, multicopy-plasmid-borne PspF or PspF delta HTH overcomes repression of the psp operon mediated by the negative regulator PspA.


[Enhancement of motor relearning and functional recovery in stroke patients: non-invasive strategies for modulating the central nervous system].
Most of the stroke survivors do not recover the basal state of the affected upper limb, suffering from a severe disability which remains during the chronic phase of the illness. This has an extremely negative impact in the quality of life of these patients. Hence, neurorehabilitation strategies aim at the minimization of the sensorimotor dysfunctions associated to stroke, by promoting neuroplasticity in the central nervous system. Brain reorganization can facilitate motor and functional recovery in stroke subjects. None-theless, after the insult, maladaptive neuroplastic changes can also happen, which may lead to the appearance of certain sensori-motor disorders such as spasticity. Noninvasive brain stimulation strategies, like transcranial direct current stimulation or transcranial magnetic stimulation, are widely used techniques that, when applied over the primary motor cortex, can modify neural networks excitability, as well as cognitive functions, both in healthy subjects and individuals with neurological disorders. Similarly, brain-machine-interface systems also have the potential to induce a brain reorganization by the contingent and simultaneous association between the brain activation and the peripheral stimulation. This review describes the positive effects of the previously mentioned neurorehabilitation strategies for the enhancement of cortical reorganization after stroke, and how they can be used to alleviate the symptoms of the spasticity syndrome.


Identifying key amino acid residues that affect α-conotoxin AuIB inhibition of α3β4 nicotinic acetylcholine receptors.
α-Conotoxin AuIB is a selective α3β4 nicotinic acetylcholine receptor (nAChR) subtype inhibitor. Its analgesic properties are believed to result from it activating GABAB receptors and subsequently inhibiting CaV2.2 voltage-gated calcium channels. The structural determinants that mediate diverging AuIB activity at these targets are unknown. We performed alanine scanning mutagenesis of AuIB and α3β4 nAChR, homology modeling, and molecular dynamics simulations to identify the structural determinants of the AuIB·α3β4 nAChR interaction. Two alanine-substituted AuIB analogues, [P6A]AuIB and [F9A]AuIB, did not inhibit the α3β4 nAChR. NMR and CD spectroscopy studies demonstrated that [F9A]AuIB retains its native globular structure, so its activity loss is probably due to loss of specific toxin-receptor residue pairwise contacts. Compared with AuIB, the concentration-response curve for inhibition of α3β4 by [F9A]AuIB shifted rightward more than 10-fold, and its subtype selectivity profile changed. Homology modeling and molecular dynamics simulations suggest that Phe-9 of AuIB interacts with a two-residue binding pocket on the β4 nAChR subunit. This hypothesis was confirmed by site-directed mutagenesis of the β4-Trp-59 and β4-Lys-61 residues of loop D, which form a putative binding pocket. AuIB analogues with Phe-9 substitutions corroborated the finding of a binding pocket on the β4 subunit and gave further insight into how AuIB Phe-9 interacts with the β4 subunit. In summary, we identified critical residues that mediate interactions between AuIB and its cognate nAChR subtype. These findings might help improve the design of analgesic conopeptides that selectively "avoid" nAChR receptors while targeting receptors involved with nociception.


Cystine uptake prevents production of hydrogen peroxide by Lactobacillus fermentum BR11.
BspA is an abundant surface protein from Lactobacillus fermentum BR11, and is required for normal cystine uptake. In previous studies, a mutant strain deficient in BspA (L. fermentum PNG201) was found to be sensitive to oxidative stress. In this study, the biochemical basis for this was explored. It was found that under aerobic batch culture conditions in de Mann-Rogosa-Sharpe medium, both L. fermentum BR11 and PNG201 entered stationary phase due to hydrogen peroxide accumulation. However, this took place at a lower optical density for PNG201 than for BR11. Measurements of hydrogen peroxide levels revealed that the BspA mutant strain overproduces this compound. Addition of 6 mM cystine to aerobic cultures was found to prevent hydrogen peroxide production by both the BR11 and PNG201 strains, but lower cystine concentrations depressed hydrogen peroxide production in BR11 more efficiently than in PNG201. Each mole of cystine was able to prevent the production of several moles of hydrogen peroxide by L. fermentum BR11, suggesting that hydrogen peroxide breakdown is dependent upon a thiol that cycles between reduced and oxidized states. It was concluded that peroxide breakdown by L. fermentum BR11 is dependent upon exogenous cystine. It is most probable that the imported L-cystine is catabolized by a cystathionine lyase and then converted into a thiol reductant for a peroxidase.


Molecular Stratification of Medulloblastoma: Clinical Outcomes and Therapeutic Interventions.
Medulloblastoma (MB) is the most common malignant pediatric posterior fossa tumor. Recent genetic, epigenetic, and transcriptomic analyses have classified MB into three subgroups, Wingless Type (WNT), Sonic Hedgehog (SHH), and non-WNT/non-SHH (originally termed Group 3 and Group 4), with discrete patient profiles and prognoses. WNT is the least common subgroup with the best prognosis, characterized by nuclear β-catenin expression, mutations in Catenin beta-1 (CTNNB1), and chromosome 6 monosomy. SHH tumors contain mutations and alterations in GLI1, GLI2, SUFU, and PTCH1 genes, which constitutively activate the SHH pathway. Originally, the presence of TP53 gene alterations and/or MYC amplifications was considered the most reliable prognostic factor. However, recent molecular analyses have subdivided SHH MB into several subtypes with distinct characteristics such as age, TP53 mutation, MYC amplification, presence of metastases, TERT promoter alterations, PTEN loss, and other chromosomal alterations as well as SHH pathway-related gene mutations. The third non-WNT/non-SHH MB (Group3/4) subgroup is genetically highly heterogeneous and displays several molecular patterns, including MYC and OTX2 amplification, GFI1B activation, KBTBD4 mutation, GFI1 rearrangement, PRDM6 enhancer hijacking, KDM6A mutation, LCA histology, chromosome 10 loss, isochromosome 17q, SNCAIP duplication, and CDK6 amplification. However, based on molecular profiling and methylation patterns, additional non-WNT/non-SHH MB subtypes have been described. Recent WHO (2021) guidelines stratified MB into four molecular subgroups with four and eight further subgroups for SHH and non-WNT/non-SHH MB, respectively. In this review, we discuss advancements in genetics, epigenetics, and transcriptomics for better characterization, prognostication, and treatment of MB using precision medicine.


[The effect of selenium therapy on autoimmune thyroiditis].
Selenium as an essential trace element is capable of exerting complex effects on the endocrine and immune system by its antioxidant capacity. The role of selenium is important because the level of free oxygen radicals is elevated in the physiological thyroid hormone synthesis. was to determine whether selenium therapy can influence the level of antithyroid peroxidase and antithyroglobulin antibodies or whether there is a correlation between antioxidant capacity and the titer of autoantibodies. 132 patient with autoimmune thyroiditis were investigated in a prospective, blind and placebo-controlled study. L-thyroxine substitution therapy was made in both groups and the level of TSH remained in the normal range. The selenium-treated group (n = 70 patients, 68 female, mean age 41,4 +/- 9,5 year) was compared with the placebo-treated group (n = 62 patients, 61 female, mean age 42,7 +/- 8,3 year). Selenium therapy was continued by L-seleno-methionine (per os 2 x 100 microg/day) for one year. Determination of TSH, fT4, fT3 and autoantibodies was carried out by chemiluminescence method. Total antioxidant capacity was determined by Randox kit, the level of selenium in the sera by atomic absorption technique was measured. In the follow-up study, patients were controlled every third month and at the end of a one-year observation period. The level of selenium in the untreated patients was significantly lower than in treated patients and controls. The fT3/fT4 ration proved to be higher in patients after selenium therapy. The titer of antithyroid antibodies (mostly the antithyroid peroxidase) significantly decreased at the end of the study. An inverse correlation was found between antioxidant capacity and the level of antithyroid peroxidase antibodies. The volume of thyroid gland slightly diminished in treated patients. Side effects were not observed. Selenium completed with L-thyroxine is a suitable therapy for patients with autoimmune thyroiditis.


De novo assembly of the transcriptome of the non-model plant Streptocarpus rexii employing a novel heuristic to recover locus-specific transcript clusters.
De novo transcriptome characterization from Next Generation Sequencing data has become an important approach in the study of non-model plants. Despite notable advances in the assembly of short reads, the clustering of transcripts into unigene-like (locus-specific) clusters remains a somewhat neglected subject. Indeed, closely related paralogous transcripts are often merged into single clusters by current approaches. Here, a novel heuristic method for locus-specific clustering is compared to that implemented in the de novo assembler Oases, using the same initial transcript collections, derived from Arabidopsis thaliana and the developmental model Streptocarpus rexii. We show that the proposed approach improves cluster specificity in the A. thaliana dataset for which the reference genome is available. Furthermore, for the S. rexii data our filtered transcript collection matches a larger number of distinct annotated loci in reference genomes than the Oases set, while containing a reduced overall number of loci. A detailed discussion of advantages and limitations of our approach in processing de novo transcriptome reconstructions is presented. The proposed method should be widely applicable to other organisms, irrespective of the transcript assembly method employed. The S. rexii transcriptome is available as a sophisticated and augmented publicly available online database.


Fluorogenic molecularly imprinted polymers with double recognition abilities synthesized via click chemistry.
This paper reports a new strategy for the preparation of molecularly imprinted polymer (MIP) based composite materials with double recognition abilities through the exploitation of click chemistry. Combining the inherent molecular recognition ability of MIPs and the affinity binding ability of boronic acid ligands for saccharides, a boronic acid-attached MIP with double recognition abilities was prepared. An alkyne modified 2,4-dichlorophenoxyacetic acid (2,4-D) imprinted polymer was first synthesized using a two-stage precipitation polymerization. An azide-contained boronic acid was then linked to the clickable 2,4-D imprinted polymers through copper-catalyzed azide-alkyne cycloaddition (CuAAC). The boronic acid-attached MIPs displayed recognition ability for 2,4-D and affinity binding ability for saccharides at physiological pH. The intensity of fluorescence emission of the boronic acid-attached MIPs was found to increase when increasing amounts of a cis-diol compound (i.e., fructose) were added.


Oncogenic mutations regulate tumor microenvironment through induction of growth factors and angiogenic mediators.
Activating mutations in the tyrosine kinase domain of HER2 (ErbB2) have been identified in human cancers. Compared with wild-type HER2, mutant HER2 shows constitutively activate kinase activity and increased oncogenicity. Cells transformed by mutant HER2 are resistant to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors and exhibit an attenuated response to the HER2 antibody trastuzumab. We investigated herein pathways through which mutant HER2 alters the extracellular environment, potentially leading to drug resistance and the effect of simultaneously targeting HER2 and the tumor cell microenvironment with a therapeutic intent. Expression of mutant HER2 in mammary epithelial cells activated autocrine transforming growth factor (TGF) beta1 signaling through a mechanism involving Rac1 and c-Jun N-terminal kinase-activating protein 1-dependent transcription. Cells transformed by an activating mutant of H-Ras (G12V) also expressed higher TGF-beta1 level through Rac1 activation. In addition, mutant HER2 induced the EGFR ligands TGF-alpha and amphiregulin at the mRNA and protein levels. Vascular endothelial growth factor, a target of the TGF-beta-Smad transcriptional regulation, was also induced as a result of expression of mutant HER2. Inhibition of TGF-beta signaling with the Alk5 small molecule inhibitor LY2109761 reduced growth and invasiveness of cells expressing mutant HER2. Combined inhibition of intracellular and paracrine effects of mutant HER2 by trastuzumab and the EGFR antibody cetuximab were more efficient than single-agent therapies. These data suggest that mutations in oncogenes such as HER2 and Ras not only alter intracellular signaling but also influence on other components of the tumor microenvironment by inducing several pro-invasive growth factors. In turn, these serve as extracellular targets of novel therapeutic strategies directed at both cancer-driving oncogenes and the modified tumor microenvironment.


Atom-specific activation in CO oxidation.
We report on atom-specific activation of CO oxidation on Ru(0001) via resonant X-ray excitation. We show that resonant 1s core-level excitation of atomically adsorbed oxygen in the co-adsorbed phase of CO and oxygen directly drives CO oxidation. We separate this direct resonant channel from indirectly driven oxidation via X-ray induced substrate heating. Based on density functional theory calculations, we identify the valence-excited state created by the Auger decay as the driving electronic state for direct CO oxidation. We utilized the fresh-slice multi-pulse mode at the Linac Coherent Light Source that provided time-overlapped and 30 fs delayed pairs of soft X-ray pulses and discuss the prospects of femtosecond X-ray pump X-ray spectroscopy probe, as well as X-ray two-pulse correlation measurements for fundamental investigations of chemical reactions via selective X-ray excitation.


Management of precocious puberty in girls with McCune-Albright syndrome using letrozole.
Objective To evaluate the safety and efficacy of letrozole in girls with progressive precocious puberty (PP) associated with McCune-Albright syndrome (MAS). Design Monocentric retrospective cross-sectional and longitudinal study of consecutive patients. Patients Ten MAS patients treated at Peking Union Medical College Hospital between September 1999 and December 2017 were retrospectively reviewed; those with complications due to PP were followed. Results The mean age at letrozole initiation was 4.5 ± 2.6 years, while the mean duration of treatment was 3.3 ± 2.4 years. Letrozole was highly effective at decreasing the rate of skeletal maturation, with a significant decrease in the bone age-to-chronological age (BA/CA) ratio from 1.9 ± 1.1 pre-treatment to 1.5 ± 1.2 on letrozole treatment (P = 0.016). Moreover, growth velocity Z-scores declined from 0.41 ± 0.5 to -0.2 ± 0.31 with treatment (P < 0.001). Predicted adult height Z-scores increased significantly from -2.03 ± 2.33 at baseline to 1.13 ± 0.84 following treatment initiation (P = 0.029). Moreover, vaginal bleeding declined significantly on letrozole. Conclusions Our findings suggest that letrozole may be an effective therapy in some girls with MAS, as treatment results in improved BA/CA ratio, growth velocity and predicted adult height. Possible adverse effects include nettle rash.


Running in place: implications of HIV incidence estimates among urban men who have sex with men in the United States and other industrialized countries.
Attempts to document changing HIV incidence rates among MSM are compromised by issues of generalizability and statistical power. To address these issues, this paper reports annualized mean HIV incidence rates from the entire published incidence literature on MSM from Europe, North America and Australia for the period 1995-2005. Publications that met the entry criteria were coded for region of the world, sampling method and year of study. From these reports, we calculated a mean incidence rate with confidence intervals for these variables. Although no differences in mean incidence rates were found for MSM from 1995 to 2005, HIV incidence rates are lower in Australia than either North America or Europe. We calculated a mean incidence rate of 2.39% for MSM in the United States, which if sustained within a cohort of MSM, would yield HIV prevalence rate of approximately 40% at age 40. These extrapolations overlap published HIV prevalence rates for MSM younger than age 40 in the United States. HIV incidence rates in the 2-3% range will adversely affect the health of gay male communities for decades to come. This analysis suggests that greater attention should be devoted to the question of how best to design prevention interventions that will lower HIV incidence rates among gay men.


The influence of abnormal hip mechanics on knee injury: a biomechanical perspective.
During the last decade, there has been a growing body of literature suggesting that proximal factors may play a contributory role with respect to knee injuries. A review of the biomechanical and clinical studies in this area indicated that impaired muscular control of the hip, pelvis, and trunk can affect tibiofemoral and patellofemoral joint kinematics and kinetics in multiple planes. In particular, there is evidence that motion impairments at the hip may underlie injuries such as anterior cruciate ligament tears, iliotibial band syndrome, and patellofemoral joint pain. In addition, the literature suggests that females may be more disposed to proximal influences than males. Based on the evidence presented as part of this clinical commentary, it can be argued that interventions which address proximal impairments may be beneficial for patients who present with various knee conditions. More specifically, a biomechanical argument can be made for the incorporation of pelvis and trunk stability, as well as dynamic hip joint control, into the design of knee rehabilitation programs. Aetiology/therapy, level 5.


Mesalazine treatment in organotypic culture of celiac patients: Comparative study with gluten free diet.
Given the central role of gluten in the pathogenesis of celiac disease (CD), a strict gluten-free diet (GFD) is the only validated treatment able to restore epithelium integrity and eliminate risks of complications. The risk of gluten contamination and the persistence of inflammation, even in patients strictly adhering to GFD, may render this treatment not always effective claiming the necessity of different new solutions. Oxidative and nitrosative stress have been indicated to play a pathophysiological role in CD. Mesalazine (5-ASA), a drug largely used in inflammatory bowel disease, has potent antinflammatory and antioxidant effects. In fact, mesalazine has been shown to decrease in vitro gluten induced cytokine response and it has been used in vivo in some refractory condition. However, its effect has never compared to that of GFD. The present study aimed to address this issue by comparing the ability of mesalazine and GFD in treating gluten-induced inflammation and oxidative stress. These effects were studied on duodenal mucosa biopsy cultures from newly diagnosed CD patients, treated or not in vitro with mesalazine, and CD biopsy cultures from patients on gluten-free diet for at least one year; and a cohort of controls constituted by healty subjects. On these models, the antioxidant cellular defences, the PPARγ, NF-kB and NOS2 proteins levels were studied. This study shows that mesalazine is as effective as GFD in reducing oxidative burst and inducing PPARγ expression; moreover it resulted more effective than GFD in decreasing NF-kB and NOS2 to the levels of controls.


Direct-calibration method for determination of aluminum in serum is comparable with the protein-precipitation technique.
We compared results by two methods for serum Al determination: matrix modification with direct calibration in a stabilized-temperature platform furnace (Clin Chem 28, 2139, 1982) and a technique involving extraction with nitric acid before atomic absorption spectrometry (Clin Chem 30, 1216, 1984). The two methods gave similar results with use of either a deuterium or a Zeeman system of background correction, but gave different slopes for standard additions (mA X s per microgram/L), depending on the Al content of the serum, an effect not seen with aqueous solutions. These differences do not affect the accuracy of the Al determination up to 150 micrograms/L.


Changes in Metabolic Profiles of Pea (Pisum sativum L.) as a Result of Repeated Short-Term Soil Drought and Subsequent Re-Watering.
The metabolic re-arrangements of peas (Pisum sativum L.) under soil drought and re-watering are still not fully explained. The search for metabolic markers of the stress response is important in breeding programs, to allow for the selection drought-resistant cultivars. During the present study, changes in the polar metabolite content in pea plant shoots were measured under repeated short-term soil drought and subsequent re-watering. A gas chromatograph, equipped with a mass spectrometer (GC-MS), was used for the metabolite profiling of pea plants during their middle stage of vegetation (14-34 days after sowing, DAS). The major changes occurred in the concentration of amino acids and some soluble carbohydrates. Among them, proline, γ-aminobutyric acid (GABA), branched-chain amino acids, hydroxyproline, serine, myo-inositol, and raffinose were accumulated under each soil drought and decreased after re-watering. Besides, the obtained results show that the first drought/re-watering cycle increased the ability of pea plants to restore a metabolic profile similar to the control after the second similar stress. The accumulation of proline seems to be an important part of drought memory in pea plants. However, confirmation of this suggestion requires metabolite profiling studies on a broader spectrum of pea cultivars.


Inverted repeats in chloroplast DNA from higher plants.
The circular chloroplast DNAs from spinach, lettuce, and corn plants have been examined by electron microscopy and shown to contain a large sequence repeated one time in reverse polarity. The inverted sequence in spinach and lettuce chloroplast DNA has been found to be 24,400 base pairs long. The inverted sequence in the corn chloroplast DNA is 22,500 base pairs long. Denaturation mapping studies have shown that the structure of the inverted sequence is highly conserved in these three plants. Pea chloroplast DNA does not contain an inverted repeat. All of the circular dimers of pea chloroplast DNA are found to be in a head-to-tail confirmation. Circular dimers of spinach and lettuce were also found to have head-to-tail conformation. However, approximately 70-80% of the circular dimers in preparations of lettuce and spinach chloroplast DNA were found to be in a head-to-head conformation. We propose that the head-to-head circular dimers are formed by a recombination event between two circular monomers in the inverted sequence.


Changes in use of types of tobacco products by pack sizes and price segments, prices paid and consumption following the introduction of plain packaging in Australia.
To describe changes among smokers in use of various types of tobacco products, reported prices paid and cigarette consumption following the standardisation of tobacco packaging in Australia. National cross-sectional telephone surveys of adult smokers were conducted from April 2012 (6 months before transition to plain packaging (PP)) to March 2014 (15 months afterwards). Multivariable logistic regression assessed changes in products, brands and pack types/sizes; multivariable linear regression examined changes in inflation-adjusted prices paid and reported cigarette consumption between the pre-PP and three subsequent periods-the transition phase, PP year 1 and PP post-tax (post a 12.5% tax increase in December 2013). The proportion of current smokers using roll-your-own (RYO) products fluctuated over the study period. Proportions using value brands of factory-made (FM) cigarettes increased from pre-PP (21.4%) to PP year 1 (25.5%; p=0.002) and PP post-tax (27.8%; p<0.001). Inflation-adjusted prices paid increased in the PP year 1 and PP post-tax phases; the largest increases were among premium FM brands, the smallest among value brands. Consumption did not change in PP year 1 among daily, regular or current smokers or among smokers of brands in any market segment. Consumption among regular smokers declined significantly in PP post-tax (mean=14.0, SE=0.33) compared to PP year 1 (mean=14.8, SE=0.17; p=0.037). Introduction of PP was associated with an increase in use of value brands, likely due to increased numbers available and smaller increases in prices for value relative to premium brands. Reported consumption declined following the December 2013 tax increase.


Aging, Diseases and X-chromatin.
X-chromatin of the buccal epithelium was determined in women aged 3-107 years. The frequency of sex chromatin occurence increase with maturation and decreases again after the menopause. The distribution of women of the child-bearing age according to X-chromatin occurence has bimodal form. With aging both modes merge. The magnitude of this change is connected with the state of health. Delayed confluence of modal variants is observed in patients with dyshormonal tumours of genital organs and the mammary gland, as well as in patients with arterial hypertension. The hypothesis that the female population consists of two subpopulations, differing by constitutional peculiarities of the endocrine regulation, is discussed.


Approaches to empiric ablation of slow pathway: results from the Canadian EP web survey.
Dual atrioventricular nodal physiology (DAVNP) is a frequent finding in patients with suspected or documented supraventricular tachycardia (SVT). Empiric slow pathway ablation (ESPA) is sometimes performed in patients with DAVNP without inducible SVT at the time of electrophysiological study. Evidence to guide this practice in the adult population is limited. This study was aimed to assess the practice of ESPA by adult electrophysiologists in Canada. All Canadian interventional electrophysiologists (n = 81) were invited to complete a web-based questionnaire assessing their practice of ESPA in patients with suspected and documented SVT. Operator experience, reimbursement models, diagnostic, and treatment decisions regarding ESPA were assessed with case scenarios. Forty-one responses (50 %) were obtained. Ninety-five percent of the responders stated that the evidence for ESPA is lacking or limited. Responders were more likely to perform ESPA in the setting of non-inducible SVT when there was documentation of the clinical arrhythmia (64 vs. 31 % (p = 0.017)). The threshold to perform ESPA was highly variable. Longer time in practice (r = 0.38, p = 0.017) and less perceived complications with ESPA (r = 0.31, p = 0.05) were correlated with the practice of ESPA, whereas length of ablation waiting lists (r = -0.15, p = 0.38), number of procedures performed per day (r = 0.11, p = 0.51) and type of reimbursement (p = 0.24) were not associated with the practice of ESPA. The perceived complication rate with ESPA was <1 %. Variability in the practice of ESPA in cases of non-inducible SVT exists. Documentation of the clinical arrhythmia, operator experience, and perceived low complication rates positively influence this practice.


Crystal structure of the cytochrome P-450CAM active site mutant Thr252Ala.
The crystal structure of a cytochrome P-450CAM site-directed mutant in which the active site Thr252 has been replaced with an Ala (Thr252Ala) has been refined to an R factor of 0.18 at 2.2 A. According to sequence alignments (Nelson & Strobel, 1989), Thr252 is highly conserved among P-450 enzymes. The crystallographic structure of ferrous camphor- and carbon monoxide-bound P-450CAM (Raag & Poulos, 1989b) suggests that Thr252 is a key active site residue, forming part of the dioxygen-binding site. Mutation of the active site threonine to alanine produces an enzyme in which substrate hydroxylation is uncoupled from electron transfer. Specifically, hydrogen peroxide and "excess" water are produced instead of the product, 5-exo-hydroxycamphor. The X-ray structure has revealed that a local distortion in the distal helix between Gly248 and Thr252 becomes even more severe in the Thr252Ala mutant. Furthermore, a solvent molecule not present in the native enzyme is positioned in the dioxygen-binding region of the mutant enzyme active site. In this location, the solvent molecule could sterically interfere with and destabilize dioxygen binding. In addition, the active site solvent molecule is connected, via a network of hydrogen bonds, with an internal solvent channel which links distal helix residues to a buried Glu side chain. Thus, solvent protons appear to be much more accessible to dioxygen in the mutant than in the wild-type enzyme, a factor which may promote hydrogen peroxide and/or water production instead of substrate hydroxylation. On the basis of crystallographic and mutagenesis data, a proton delivery pathway involving residues Lys178/Arg186, Asp251, and Thr252 is proposed for wild-type P-450CAM. Coordinates of structures discussed in this paper have been submitted to the Brookhaven Protein Data Bank (Bernstein et al., 1977).


Smartphone-Guided Educational Counseling and Self-Help for Chronic Tinnitus.
Tinnitus is an auditory phantom perception in the ears or head in the absence of a corresponding external stimulus. There is currently no effective treatment available that reliably reduces tinnitus. Educational counseling is a treatment approach that aims to educate patients and inform them about possible coping strategies. For this feasibility study, we implemented educational material and self-help advice in a smartphone app. Participants used the educational smartphone app unsupervised during their daily routine over a period of four months. Comparing the tinnitus outcome measures before and after smartphone-guided treatment, we measured changes in tinnitus-related distress, but not in tinnitus loudness. Improvements on the Tinnitus Severity numeric rating scale reached an effect size of 0.408, while the improvements on the Tinnitus Handicap Inventory (THI) were much smaller with an effect size of 0.168. An analysis of user behavior showed that frequent and intensive use of the app is a crucial factor for treatment success: participants that used the app more often and interacted with the app intensively reported a stronger improvement in the tinnitus. Between study allocation and final assessment, 26 of 52 participants dropped out of the study. Reasons for the dropouts and lessons for future studies are discussed in this paper.


Photooxidation of cyanobacteria in natural conditions.
Photodynamic effects were demonstrated and assayed under field conditions in a number of different laboratory strains and pond isolates of cyanobacteria; parameters assayed for resistance to photooxidation were viable count, turbidity of the cyanobacterial suspension, and protein and pigment contents. The effects of density, colonial structure, and internal gas vacuoles on the lethal outcome were investigated. The stability and formation of superoxide dismutase under photooxidative conditions in the field and laboratory were studied in the different strains. An isolate of Microcystis from blooms in ponds exhibited extremely high resistance to photooxidation, which was abolished by exposure to chloramphenicol.


Hemodynamic studies in spontaneously hypertensive rats with congenital arteriovenous shunts.
Previous studies have demonstrated a subset of normotensive rats (Wistar-Kyoto, WKY) that has arteriovenous shunts and biventricular hypertrophy. In this study, using the combined radioactive reference sample and microsphere techniques, spontaneously hypertensive (SHR) and WKY rats were found to have arteriovenous fistulas, and the magnitude of this shunting was quantified. In both groups with shunts (WKY and SHR, respectively) mean arterial pressure was lower (117 +/- 3 vs. 125 +/- 2 mmHg, NS; 164 +/- 4 vs. 184 +/- 7 mmHg, P less than 0.001) and cardiac index was higher (518 +/- 52 vs. 259 +/- 10 ml.min-1.kg-1, P less than 0.001: 416 +/- 18 vs. 239 +/- 8 ml.min-1.kg-1, P less than 0.001) than in rats without shunts. Both groups demonstrated sizable shunts, with microsphere trapping in 55 +/- 6 and 47 +/- 5% in WKY and SHR, respectively, and reduced blood flow to every organ. These data demonstrate the need for caution in evaluating hemodynamic and myocardial functional data as well as endocrinologic or metabolic functions in WKY and SHR unless shunting is excluded. Moreover, since these subsets seem to have had arteriovenous shunts from birth, it may be inferred that the development of the arterial hypertension in SHR did not result from tissue hyperperfusion.


Three new cardenolides from the fruits of Cascabela thevetia (L.) Lippold and their cytotoxic activities.
Phytochemical investigations on the fruits of Cascabela thevetia (L.) Lippold led to obtain three new cardenolides (1-3) and five known analogues (4-7). Their structures were elucidated by means of UV, IR, HR-ESI-MS, 1D and 2D NMR spectroscopic data analysis. Compounds 1 and 2 represent the first examples of naturally occurring cardenolides with 19-nor-5(10)-ene group and α-l-3-demethyl-thevetose, respectively. Compound 3 is a rare C-nor-D-homocardenolide in nature. All isolated cardenolides (1-7) were evaluated for their cytotoxic activities against four human cancer cell lines (MCF-7, HCT-116, HeLa and HepG2), and the results indicated the compounds with sugar units (1, 2, 4, and 5) exhibited stronger cytotoxic activities with IC50 values ranging between 0.022 and 0.308 μM.


Brightly luminescent organically capped silicon nanocrystals fabricated at room temperature and atmospheric pressure.
Silicon nanocrystals are an extensively studied light-emitting material due to their inherent biocompatibility and compatibility with silicon-based technology. Although they might seem to fall behind their rival, namely, direct band gap based semiconductor nanocrystals, when it comes to the emission of light, room for improvement still lies in the exploitation of various surface passivations. In this paper, we report on an original way, taking place at room temperature and ambient pressure, to replace the silicon oxide shell of luminescent Si nanocrystals with capping involving organic residues. The modification of surface passivation is evidenced by both Fourier transform infrared spectroscopy and nuclear magnetic resonance measurements. In addition, single-nanocrystal spectroscopy reveals the occurrence of a systematic fine structure in the emission single spectra, which is connected with an intrinsic property of small nanocrystals since a very similar structure has recently been observed in specially passivated semiconductor CdZnSe nanoparticles. The organic capping also dramatically changes optical properties of Si nanocrystals (resulting ensemble photoluminescence quantum efficiency 20%, does not deteriorate, radiative lifetime 10 ns at 550 nm at room temperature). Optically clear colloidal dispersion of these nanocrystals thus exhibits properties fully comparable with direct band gap semiconductor nanoparticles.


Hybro chicks outperform Ross308 in a numerical-ordinal task. Cognitive and behavioral comparisons between 2 broiler strains of newborn domestic chicks (Gallus gallus).
Domestic chickens (Gallus gallus) are among those species subject to intensive selection for production. Among the most widely used broiler strains are the Ross308 and the Hybro. From the perspective of animal production, Ross308 were superior to Hybro in weight gain, final body mass, and feed conversion. Intensive selection is thought to also cause behavioral changes and to negatively affect cognitive abilities. Up to date, though, no evidence has been provided on broiler breeds. The aim of this study was to explore cognitive differences among Hybro and Ross308 chickens by assessing their ordinal-numerical abilities. Chicks learned learnt to find a food reward in the 4th container in a series of 10 identical and sagittally aligned containers. We designed a standard training procedure ensuring that all chicks received the same amount of training. The chicks underwent 2 tests: a sagittal and a fronto-parallel one. In the former test, the series was identical to that experienced during training. In the fronto-parallel test, the series was rotated by 90°, thus left-to-right oriented, to assess the capability of transferring the learnt rule with a novel spatial orientation. In the sagittal test, both chicken hybrids selected the 4th item above chance; interestingly the Hybro outperformed the Ross308 chicks. In the fronto-parallel test, both strains selected the 4th left and the 4th right container above chance; nevertheless, the Hybro chicks were more accurate. Our results support the hypothesis that intense selection for production can influence animal cognition and behavior, with implications on animal husbandry and welfare.


Gerodontology--how big is the challenge in Ireland?
Population trends suggest that the Irish population is ageing, and that this population will have substantial treatment needs. These patients will be better informed than previous generations, and will demand treatment aimed at preserving a natural dentition. This will impact upon delivery of oral healthcare and manpower planning needs to consider how to address the increased demand for dental care. Poor oral health is associated with systemic health problems, including cardiovascular disease, respiratory disease and diabetes mellitus. It also has a negative impact upon quality of life, and the World Health Organisation has encouraged public healthcare administrators and decision makers to design effective and affordable strategies for better oral health and quality of life of older adults, which, in turn, are integrated into general health management programmes. Treatment concepts such as minimally invasive dentistry and the shortened dental arch concept are discussed in the context of these demographic changes and recommendations.


Mechanism of carbon monoxide oxidation by the carbon monoxide dehydrogenase/acetyl-CoA synthase from Clostridium thermoaceticum: kinetic characterization of the intermediates.
Carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) from Clostridium thermoaceticum catalyzes (i) the synthesis of acetyl-CoA from a methylated corrinoid protein, CO, and coenzyme A and (ii) the oxidation of CO to CO2. CO oxidation occurs at a Ni- and FeS-containing center known as cluster C. Electrons are transferred from cluster C to a separate metal center, cluster B, to external acceptors like ferredoxin. In the work described here, we performed reductive titrations of CODH/ACS with CO and sodium dithionite and monitored the reaction by electron paramagnetic resonance (EPR) spectroscopy. We also performed pre-steady-state kinetic studies by rapid freeze-quench EPR spectroscopy (FQ-EPR) and stopped-flow kinetics. Redox titrations of CODH/ACS revealed the existence of a UV-visible and EPR-silent electron acceptor denoted center S that does not appear to be associated with any of the other metal centers in the protein. Our results support the previous proposals [Anderson, M. E., & Lindahl, P. A. (1994) Biochemistry 33, 8702-8711; Anderson, M. E., & Lindahl, P. A. (1996) Biochemistry 35, 8371-8380] that the Cred2 form of cluster C is two electrons more reduced than the Cred1 form. The combined results from titrations and pre-steady-state studies were used to formulate a mechanism for CO oxidation, composed of the following steps: (i) CO binding to the [Cred1,Box, Xox] state to yield a Cred1-CO complex; (ii) two-electron reduction of Cred1 to Cred2 concerted with CO2 release; (iii) binding of a second CO molecule to the [Cred2,Box,Xox] state to form a Cred2-CO complex; (iv) electron transfer from Cred2-CO to cluster B to form [Cred2,Bred,Xred] with concerted release of the second CO2. Step iii competes with internal electron transfer from Cred2 to Box and Xox. At high CO concentrations, step iii is favored, whereas at low concentrations, only one CO molecule per turnover binds and undergoes oxidation. Closure of the catalytic cycle involves electron transfer from reduced enzyme to an electron acceptor protein, like ferredoxin. Xox is a yet-uncharacterized electron acceptor that may be an intermediate in the reduction of center S. The Cred2 state appears to be the predominant state of cluster C during steady-state turnover. The rate-determining step for the first half-reaction is step iv, while during steady-state turnover, it appears to be electron transfer to external electron acceptors.


Effect of calorie restriction or protein intake on circulating levels of insulin like growth factor I in humans: A systematic review and meta-analysis.
Calorie restriction (CR) and reductions in protein intake in rodents result in increased lifespan and reduced levels of IGF-1. However, the changes in IGF-1 in humans in response to CR and elevated protein intake are confused. We conducted a systematic review and meta-analysis to investigate the effect of Calorie restriction (CR) or increase in protein intake on IGF-1 in humans. The systematic review protocols have been developed based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement guidelines. Two separate systematic searches were undertaken: first for the effect of CR and second on the effect of increase in protein intake on IGF-1. PubMed, SCOPUS and ISI Web of Science databases were searched. In the meta-analysis of the calorie restriction studies, twelve studies met the inclusion criteria (8 clinical trials and 4 observational studies). The meta-analysis of both clinical trials and observational studies revealed no significant effect of CR on IGF-1 (clinical trials: standardized mean difference (SMD) = 0.002 ng/ml, 95% CI -0.14 to 0.14 ng/ml, p = 0.98; observational studies (SMD = -1.14 ng/ml, 95% CI -1.9 to -0.38 ng/ml, p = 0.003). In the meta-analysis of protein intake studies (six studies), a significant increase in circulating IGF-1 levels in response to increases in dietary protein was revealed (SMD = 0.4 ng/ml, 95% CI 0.18-0.61 ng/ml, p < 0.001). In conclusion, in humans, CR was not associated with a significant change in circulating IGF-1. However an increase in protein intake was associated with increased levels of circulating IGF-1. PROTOCOLS REGISTRATION NUMBER: CRD42017073149 for the protein intake meta-analysis and CRD42016046260 for CR meta-analysis.


Milk production response to diets supplemented with dried brewers grains, wet brewers grains, or soybean meal.
Forty four multiparous Holsteins, 114 +/- 28 days in milk (32 kg/day, 3.4% fat), were fed a basal diet of 12% crude protein and 20% acid detergent fiber for 10 days, then stratified by milk production into four groups. Cows were randomized to one of nine diets in a 3 X 3 factorial in which basal was supplemented with dried brewers grains, wet brewers grains, or soybean meal to supply in the diet 14.5, 16.0, and 17.5% crude protein. Eight cows remained on basal during the 50-day trial. Basal contained: 14% alfalfa silage, 27% ensiled ground-corn, 53% corn silage, and 6% vitamin-mineral mix. Milk production (kg/day) for cows fed dried brewers grains (29.4) and wet brewers grains (28.9) was higher than soybean meal (26.2) and basal (23.1). Milk production was different for diets with high (29.6) vs. low (27.8) and medium (27.2) protein. Dry matter intake (as percent of body weight) was 3.7, 3.5, 3.3, and 2.9 for dried brewers, wet brewers, soybean meal, and basal, respectively. Milk protein percent and milk fat percent differed for protein source. Rumen fluid ammonia nitrogen for combined 2, 4, and 6 h post-feeding, was (mg/100 ml) 10.4 for dried brewers, 14.9 for wet brewers, and 18.0 for soybean meal and increased from 13.2 to 15.4 with increased protein. Plasma urea tended to follow patterns of rumen ammonia. Dried brewers grains had lower apparent nitrogen digestibility but equal nitrogen balance, indicating more efficient metabolic use than soybean meal.


"Door-In to Door-Out" Delay in Patients with Acute ST-Segment Elevation Myocardial Infarction Transferred for Primary Percutaneous Coronary Intervention in a Metropolitan STEMI Network of a Developing Country.
Background Routine performance measures of primary percutaneous coronary intervention (PCI) within an ST-segment elevation myocardial infarction (STEMI) network are needed to improve care. Objective We evaluated the door-in to door-out (DI-DO) delays at the initial hospitals in STEMI patients as a routine performance measure of the metropolitan STEMI network. Patients and Methods We retrospectively analyzed the DI-DO time from 1,076 patients with acute STEMI who were transferred by ground ambulance to a primary PCI center for primary PCI between 4 October 2014 and 1 April 2019. Correlation analysis between DI-DO times and total ischemia time was performed using Spearman's test. Logistic regression analyses were used to find variables associated with a longer DI-DO time. Results Median DI-DO time was 180 minutes (25th percentile to 75th percentile: 120-252 minutes). DI-DO time showed a positive correlation with total ischemia time ( r = 0.4, p < 0.001). The median door-to-device time at the PCI center was 70 minutes (25th percentile to 75th percentile: 58-88 minutes). Multivariate analysis showed that women patients were independently associated with DI-DO time > 120 minutes (odds ratio 1.55, 95% confidence interval 1.03 to 2.33, p = 0.03). Conclusion The DI-DO time reported in this study has not reached the guideline recommendation. To improve the overall performance of primary PCI in the region, interventions aimed at improving the DI-DO time at the initial hospitals and specific threat for women patients with STEMI are possibly the best efforts in improving the total ischemia time.


Pancreas transplantation in The Netherlands: report of the first case.
This case report describes the first clinical pancreas transplant performed in The Netherlands. A duct-obliterated segmental graft from a cadaver donor was transplanted to a male recipient who was insulin-dependent after total pancreatectomy eight years earlier. The recipient was off insulin for four postoperative months; during this period a rejection crisis was successfully treated. The graft was removed eight months after transplantation. Histologic examination indicated both fibrosis and rejection to have presumably contributed to graft failure. The patient is well but again insulin-dependent.


Dental and Oral Lesions in HIV-Positive Individuals in East Bohemia--Czech Republic, Single Centre Experience.
Human immunodeficiency virus (HIV) continues to be a serious health issue and one of the world most devastating epidemics. An estimated 1.5 million people died from AIDS-related illnesses in 2013, and an estimated 37 million people with AIDS have died worldwide since the epidemic has begun. HIV infection is known for its oral manifestations which causes discomfort and pain for infected individuals. The objective of this study was to document oral conditions of HIV positive patients and the pattern and frequency of oral and dental lesions. All patients with confirmed HIV infection who were treated at the Department of Dentistry, University Hospital in Hradec Králové, were examined. During the study period, 29 HIV positive patients were examined and treated--19 men, 10 women, with mean age of 32.9 years (range 22-58 years). 72.41% patients received ART. In total, all patients underwent 186 visits. The most frequent treatments were associated with teeth and periodontal lesions (71.80%), oral mucosal lesions were diagnosed and treated only in 3.96% cases. Since the introduction of ART, the frequency of oral mucosal lesions is minimal in patients with HIV infection.