Effect of purified staphylococcal leukocidal toxins on isolated blood polymorphonuclear leukocytes and peritoneal macrophages in vitro.
Bicomponent (fractions S and F) staphylococcal leukocidal toxins (Panton-Valentine leukocidin-Luk and haemolysin gamma-Hlg) were tested for in vitro activity against isolated polymorphonuclear leukocytes (PMNL) and peritoneal macrophages (PMF). For assessment of membrane permeability at subcytolytic concentrations of leukocidin (Luk-S + Luk-F) and haemolysin gamma (HlgA + HlgB) (8-1000 ng/ml), PMNL and PMF were radiolabelled (86Rb, 14C-amino-isobutyric acid (AIB) or 51Cr). All toxins tested caused lysis of human PMNL, although considerable differences were noted in the sensitivity of these cells to Luk and Hlg. Release of 51Cr (at 1000-5000 ng/ml), being a sign of irreversible cell damage and lysis, was preceded, at lower concentrations of the toxins (40 and 200 ng/ml), by the release of large amounts of low-molecular labels--86Rb and 14C-AIB. In another experiment, it was found that release of 86Rb from PMNL incubated with low concentrations of Luk (50 ng/ml) took place after 15-30 minutes of incubation, when no significant amounts of 14C-AIB or 51Cr were released. These findings support the concept of pore formation by staphylococcal leukocidal toxins in membranes of sensitive cells and indicate that a relatively short time is needed for the formation of these pores after binding of the Luk-S and Luk-F components to the membrane.


Acceleration of Gastric Emptying by Insulin-Induced Hypoglycemia is Dependent on the Degree of Hypoglycemia.
Hypoglycemia is a major barrier to optimal glycemic control in insulin-treated diabetes. Recent guidelines from the American Diabetes Association have subcategorized "non-severe" hypoglycemia into level 1 (<3.9 mmol/L) and 2 (<3 mmol/L) hypoglycemia. Gastric emptying of carbohydrate is a major determinant of postprandial glycemia but its role in hypoglycemia counter-regulation remains underappreciated. "Marked" hypoglycemia (~2.6 mmol/L) accelerates gastric emptying and increases carbohydrate absorption in health and type 1 diabetes, but the impact of "mild" hypoglycemia (3.0-3.9 mmol/L) is unknown. To determine the effects of 2 levels of hypoglycemia, 2.6 mmol/L ("marked") and 3.6 mmol/L ("mild"), on gastric emptying in health. Fourteen healthy male participants (mean age: 32.9 ± 8.3 years; body mass index: 24.5 ± 3.4 kg/m2) from the general community underwent measurement of gastric emptying of a radiolabeled solid meal (100 g beef) by scintigraphy over 120 minutes on 3 separate occasions, while blood glucose was maintained at either ~2.6 mmol/L, ~3.6 mmol/L, or ~6 mmol/L in random order from 15 minutes before until 60 minutes after meal ingestion using glucose-insulin clamp. Blood glucose was then maintained at 6 mmol/L from 60 to 120 minutes on all days. Gastric emptying was accelerated during both mild (P = 0.011) and marked (P = 0.001) hypoglycemia when compared to euglycemia, and was more rapid during marked compared with mild hypoglycemia (P = 0.008). Hypoglycemia-induced gastric emptying acceleration during mild (r = 0.57, P = 0.030) and marked (r = 0.76, P = 0.0014) hypoglycemia was related to gastric emptying during euglycemia. In health, acceleration of gastric emptying by insulin-induced hypoglycemia is dependent on the degree of hypoglycemia and baseline rate of emptying.


Global prevalence of diabetes in active tuberculosis: a systematic review and meta-analysis of data from 2·3 million patients with tuberculosis.
Although diabetes and poor glycaemic control significantly increase the risk of tuberculosis and adversely affect tuberculosis treatment outcomes, the global burden of diabetes in the context of tuberculosis remains unknown. We did a systematic review and meta-analysis to estimate the prevalence of diabetes among patients with tuberculosis at global, regional, and country levels. We searched PubMed, Excerpta Medica Database, Web of Science, and Global Index Medicus to identify studies published between Jan 1, 1986, and June 15, 2017, on the prevalence of diabetes in patients with active tuberculosis, with no language restrictions. Criteria to diagnose tuberculosis and diabetes concurred with WHO guidelines. Methodological quality of eligible studies was assessed, and random-effect models meta-analysis served to obtain the pooled prevalence estimate of diabetes among patients with active tuberculosis, globally. Heterogeneity (I2) was assessed via the χ2 test on Cochran's Q statistic. This study is registered with PROSPERO, number CRD42016049901. We screened 7565 records of which 200 studies (2 291 571 people with active tuberculosis) were included in meta-analyses. The pooled prevalence of diabetes was 15·3% (95% prediction interval 2·5-36·1; I2 99·8%), varying from 0·1% in Latvia to 45·2% in Marshall Islands. Subgroup and metaregression analyses for identifying sources of heterogeneity showed that four International Diabetes Federation (IDF) regions (North America and Caribbean [19·7%], western Pacific [19·4%], southeast Asia [19·0%], Middle East and North Africa [17·5%]) had significantly higher prevalence estimates than the three others (Africa [8·0%], South and Central America [7·7%], and Europe [7·5%]; p<0·0001). Additionally, the prevalence increased with age, in men, and in countries with low tuberculosis burden. The prevalence of diabetes was decreased in countries that had low incomes and low Human Development Index scores. The form of tuberculosis infection and presence of HIV seemed not to affect the prevalence of diabetes among patients with active tuberculosis. This study suggests a high burden of diabetes among patients with active tuberculosis, with disparities according to age, sex, regions, level of country income, and development. Cost-effective strategies to curb the burden of diabetes among patients with active tuberculosis are needed. None.


Role of local treatment including radiotherapy in Barcelona Clinic of Liver Cancer stage C patients: a nationwide cohort analysis in South Korea.
Sorafenib is recommended as the standard treatment for hepatocellular carcinoma (HCC) of Barcelona Clinic of Liver Cancer stage C (BCLC C). However, local treatment including radiation therapy (LRT) is also widely administered in practice. The aim of our study was to define the role of LRT among BCLC C patients. Of the patients with HCC enrolled the Korean Central Cancer Registry from 2008 to 2014, the Korean Liver Cancer Study Group randomly extracted 10,580 patient data from ~50 hospitals nationwide. Among them, 3,401 patients were categorized to have BCLC C HCC. Among them, patients with information on initial treatments, defined as the first and secondary treatment within 60 days after the first treatment, were selected and classified into three initial treatment groups: LRT, sorafenib, and no treatment. Among 3,401 BCLC C HCC patients, 1,486 were included in the study and the remaining patients were excluded as they did not meet the criteria (eg, underwent local treatments without radiotherapy [RT] or received chemotherapy other than sorafenib). Of these, 266 were assigned to LRT (17.9%), 316 to sorafenib (21.3%), and 904 to no treatment group (60.8%). Median survival time of the sorafenib group was shorter than that of the LRT group (3.8 vs 7.6 months, P<0.001). In multivariable analysis, sorafenib group showed significantly higher risk related to mortality compared to LRT group, not only among all patients (HR: 1.50, 95% CI: 1.23-1.84) but also between subgroup cohorts with portal invasions (1.55, 1.23-1.84), with lymph node metastases (2.42, 1.53-3.83), without distant metastases (1.43, 1.10-1.87), and with distant metastases (1.57, 1.13-2.19). Additionally, no treatment group showed the worst survival among the three treatment groups not only in all patients, but also in all subgroups of patients (P<0.001 in all). LRT as an initial treatment showed survival benefit as compared to sorafenib in HCC patients of BCLC C.


Translational reprogramming as a driver of antimony-drug resistance in Leishmania.
Leishmania is a unicellular protozoan that has a limited transcriptional control and mostly uses post-transcriptional regulation of gene expression, although the molecular mechanisms of the process are still poorly understood. Treatments of leishmaniasis, pathologies associated with Leishmania infections, are limited due to drug resistance. Here, we report dramatic differences in mRNA translation in antimony drug-resistant and sensitive strains at the full translatome level. The major differences (2431 differentially translated transcripts) were demonstrated in the absence of the drug pressure supporting that complex preemptive adaptations are needed to efficiently compensate for the loss of biological fitness once they are exposed to the antimony. In contrast, drug-resistant parasites exposed to antimony activated a highly selective translation of only 156 transcripts. This selective mRNA translation is associated with surface protein rearrangement, optimized energy metabolism, amastins upregulation, and improved antioxidant response. We propose a novel model that establishes translational control as a major driver of antimony-resistant phenotypes in Leishmania.


Germ-line and somatic PTPN11 mutations in human disease.
Reversible protein tyrosyl phosphorylation of cell surface receptors and downstream intracellular transducers is a major regulatory mechanism used to modulate cellular responses to extracellular stimuli, and its deregulation frequently drives aberrant cell proliferation, survival and/or differentiation. SHP-2 is a cytoplasmic Src-homology 2 domain-containing protein tyrosine phosphatase that plays an important role in intracellular signaling and is required during development and hematopoiesis. Germ-line missense mutations in PTPN11, the gene coding SHP-2, have been discovered as a major molecular event underlying Noonan syndrome, an autosomal dominant trait characterized by short stature, dysmorphic facies, and congenital heart defects, as well as in other closely related developmental disorders. More recently, a distinct class of missense mutations in the same gene has been identified to occur as a somatic event contributing to myeloid and lymphoid malignancies. This review focuses on the role of SHP-2 in signal transduction, development and hematopoiesis, as well as on the consequences of SHP-2 gain-of-function.


[Laparoscopic extraperitoneal adenomectomy: surgical technique and preliminary results].
We analyze the laparascopic adenomectomy preliminary results and describe the surgical technique, for benign prostatic hyperplasia (BPH), for glands greater than 60cc. From January to April 2009 we have performed 10 laparoscopic adenomectomies. We performed a descriptive and retrospective study and early postoperative results were analyzed. Descriptive statistical analyses were performed using IPSS 17.0. The surgical technique is described step by step. Conversion to open surgery was not required, and none of the patients had serious peri-operative and post-operative complications. None of the ten patients required blood transfusions. Median operating time was 112,5min (80-135). Median hospital stay was 3,5 days (2-5) and median catheterization period was 7 days (3-21). The median prostate enucleated weight was 62gr. (40-93). The median postoperative Qmax was 18,8ml/seg and the median score of IPSS was 5. Laparoscopic adenomectomy is a low rate morbidity technique and reproducible in centers with laparoscopic skills. Prospective and comparative studies with open surgery will be necessary to choose the best technique for our patients.


Bright and Stable Quantum Dot Light-Emitting Diodes.
Quantum dot light-emitting diodes (QLEDs) are one of the most promising candidates for next-generation displays and lighting sources, but they are barely used because vulnerability to electrical and thermal stresses precludes high brightness, efficiency, and stability at high current density (J) regimes. Here, bright and stable QLEDs on a Si substrate are demonstrated, expanding their potential application boundary over the present art. First, a tailored interface is granted to the quantum dots, maximizing the quantum yield and mitigating nonradiative Auger decay of the multiexcitons generated at high-J regimes. Second, a heat-endurable, top-emission device architecture is employed and optimized based on optical simulation to enhance the light outcoupling efficiency. The multilateral approaches realize that the red top-emitting QLEDs exhibit a maximum luminance of 3 300 000 cd m-2 , a current efficiency of 75.6 cd A-1 , and an operational lifetime of 125 000 000 h at an initial brightness of 100 cd m-2 , which are the highest of the values reported so far.


Design and synthesis of artificial phospholipid for selective cleavage of integral membrane protein.
An artificial phospholipid, possessing saturated alkyl chains as a membrane anchor and protein recognition site as well as an Fe(III)-EDTA moiety as a protein cleavable polar head group, was designed and synthesized based on the amidite method for the purpose of examination of cleavage of integral membrane proteins.


N6-methyldeoxyadenosine marks active transcription start sites in Chlamydomonas.
N(6)-methyldeoxyadenosine (6mA or m(6)A) is a DNA modification preserved in prokaryotes to eukaryotes. It is widespread in bacteria and functions in DNA mismatch repair, chromosome segregation, and virulence regulation. In contrast, the distribution and function of 6mA in eukaryotes have been unclear. Here, we present a comprehensive analysis of the 6mA landscape in the genome of Chlamydomonas using new sequencing approaches. We identified the 6mA modification in 84% of genes in Chlamydomonas. We found that 6mA mainly locates at ApT dinucleotides around transcription start sites (TSS) with a bimodal distribution and appears to mark active genes. A periodic pattern of 6mA deposition was also observed at base resolution, which is associated with nucleosome distribution near the TSS, suggesting a possible role in nucleosome positioning. The new genome-wide mapping of 6mA and its unique distribution in the Chlamydomonas genome suggest potential regulatory roles of 6mA in gene expression in eukaryotic organisms.


Patterns from dried drops as a characterisation and healthcare diagnosis technique, potential and challenges: A review.
When particulate-laden droplets evaporate, they leave behind complex patterns on the substrate depending on their composition and the dynamics of their evaporation. Over the past two decades, there has been an increased interest in interpreting these patterns due to their numerous applications in biomedicine, forensics, food quality analysis and inkjet printing. The objective of this review is to investigate the use of patterns from dried drops as a characterisation and diagnosis technique. The patterns left behind by dried drops of various complex fluids are categorised. The potential applications of these patterns are presented, focussing primarily on healthcare, where the future impact could be greatest. A discussion on the limitations which must be overcome and prospective works that may be carried out to allow for widespread implementation of this technique is presented in conclusion.


[Adjuvant whole body acupuncture in depression. A placebo-controlled study with standardized mianserin therapy].
In order to examine the efficacy of whole body acupuncture additionally applied to drug treatment in depression, a single-blind placebo-controlled study with 70 inpatients administered to three different treatment groups has been carried out. All patients were pharmacologically treated with the tetracyclic antidepressant mianserin. The verum group (n = 22) received acupuncture at specific points considered to be effective in the treatment of depression. The placebo group (n = 24) was treated with acupuncture at non-specific locations and the control group (n = 24) received only pharmacological treatment. Acupuncture was applied three times a week over a period of four weeks. Psychopathology was rated by judges blind to verum/placebo conditions twice a week over eight weeks with the CGI, GAS, BRMS and BfS rating scales. Additionally applied acupuncture improved the course of depression more than pharmacological treatment with mianserin did by itself. However, we could not detect any differences between placebo and verum acupuncture.


Genetic Basis of Posterior Urethral Valves Inheritance.
To highlight genetic pattern of posterior urethral valves (PUVs), we performed a genetic study on 2 siblings affected. PUVs are the most common congenital cause of lower urinary tract obstruction and an important cause of renal failure in infants (50% progress to end-stage renal disease in 10 years). PUVs occur in 1 of 5000-8000 male infants, but real incidence is arduous to determine because of the wide spectrum of possible clinical presentation. A different recurrence rate is reported in African Americans and children with Down syndrome, although usually PUVs are not found in syndromic conditions but constitute an isolated disorder. Although most cases appear to be sporadic, some reports in literature suggest a partial genetic etiology. We report 2 brothers with PUVs. The children's mother was a healthy woman but had a history of urinary tract infections of unknown etiology. We investigated possible familial genetic anomalies using a DNA array comparative genomic hybridization technique. We identified 2 partial duplications in the short arm of chromosome 11 recurring in both children and mother. This finding, not previously reported to our knowledge, adds new data to support the hypothesis of the presence of a hereditary component in the occurrences of PUVs.


Inhibitory effects of pectenotoxins from marine algae on the polymerization of various actin isoforms.
Pectenotoxins (PTXs) are marine toxins produced by dinoflagellates and which accumulate in shellfish. There are at least 14 different analogs of PTX with slight variations in structure leading to different chemical properties and consequently different toxicities. Since preliminary studies have shown that the parent compound PTX1 targets actin, we investigated the effects of two analogs, PTX2 and PTX2 seco acid, on the polymerization and depolymerization of skeletal muscle actin, smooth muscle actin, cardiac muscle actin, and non-muscle actin. Optimized actin assays using fluorescently labeled skeletal muscle actin and SDS-PAGE were jointly used to determine the relative amounts of filamentous and globular actin formed during polymerization and depolymerization experiments. Our findings suggest that PTX2 causes a dose-dependent decrease in both the rate and yield of skeletal muscle actin polymerization (IC50 values of 44 and 177 nM; respectively), with no significant effects on depolymerization. Moreover, the inhibitory effects of PTX2 are conserved towards other actin isoforms (i.e., smooth muscle, cardiac muscle, and non-muscle), as the inhibitory effects on actin polymerization were also observed with similar IC50 values (range: 19-94 nM). No inhibitory effects on polymerization were observed for PTX2 seco acid, suggesting an intact lactone ring is necessary for bioactivity.


Dephosphorylation of Six2Y129 protects tyrosine hydroxylase-positive cells in SNpc by regulating TEA domain 1 expression.
Parkinson's disease (PD) is a neurodegenerative disease characterized by selective loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). We recently reported that Six2 could reverse the degeneration of DA neurons in a dephosphorylation state. Here we further identified that Eya1 was the phosphatase of Six2 that could dephosphorylate the tyrosine 129 (Y129) site by forming a complex with Six2 in damaged DA cells. Dephosphorylated Six2 then translocates from the cytoplasm to the nucleus. Using ChIP-qPCR and dual luciferase assay, we found that dephosphorylated Six2 down-regulates TEA domain1 (Tead1) expression, thus inhibiting 6-hydroxydopamine (6-OHDA)-induced apoptosis in DA cells. Furthermore, we showed Six2Y129F/Tead1 signaling could protect against the loss of SNpc tyrosine hydroxylase-positive (TH+) cells and improve motor function in PD model rats. Our results demonstrate a dephosphorylation-dependent mechanism of Six2 that restores the degeneration of DA neurons, which could represent a potential therapeutic target for PD.


Regional odontodysplasia.
Regional odontodysplasia was diagnosed in a girl aged 6 with lesions of a number of deciduous and permanent teeth in the right lower and upper quadrants. The literature comprises reports on some 50 cases of regional odontodysplasia, including only one patient who also showed involvement of both the maxilla and the mandible.


Accelerated polyethylene wear and early failure of the uncemented Reflection acetabular component: a ten to fifteen-year follow-up study.
The Reflection® second-generation uncemented acetabular component was designed to address increased rates of failure observed with the early acetabular designs. However, the reported survivorship of this acetabular component has been conflicting. The aim of this study was to describe the ten to 15-year survivorship and polyethylene wear rate for the uncemented Reflection® acetabular component performed as part of a primary total hip replacement. One hundred and four consecutive Reflection® uncemented acetabular components in 97 patients were identified from a prospective arthroplasty database with a minimum of ten years of follow-up. No patient was lost to follow-up. Mean cohort age was 59.1 years. There were 24 revisions of the acetabular component. The all-cause survival rate at ten years was 77.2% [95% confidence interval (CI) 73.9-80.5]. Mean linear wear was 0.20 mm [standard deviation (SD) 0.08] per year and the volumetric wear was 106.2 mm(3) per year. At final follow-up, more than half the patients had osteolysis identified around the femoral component. The mean Oxford Hip Score was 35.6 (SD 9.8) at a mean follow-up of 11.9 years, with six (11.1%) excellent, 26 (48.1%) good, 17 (31.5%) fair and five (9.3%) poor outcomes. Despite the fact that more than a third had a fair or poor outcome, only four (7.4%) were not satisfied with their hip. Due to the high rate of relatively asymptomatic polyethylene wear and osteolysis associated with this acetabular component, in our department, we now review all surviving patients both clinically and radiographically on an annual basis.


Ectopic soft tissue calcium deposition following liver transplantation.
We observed ectopic soft tissue calcification affecting seven patients following orthotopic liver transplantation. The cause of such calcification is unknown, but potential pathogenetic factors include hyperparathyroidism, calcium administered during and following surgery, renal failure, acid-base changes and citrate in fresh frozen plasma. To investigate some of the mechanisms underlying ectopic calcification following liver transplantation, we determined preoperative levels of ionized serum calcium, phosphate, magnesium, parathyroid hormone (midmolecule assay) and 1,25-(OH)2 vitamin D in 20 patients who underwent 24 liver transplants. In addition, these parameters were measured weekly in 15 patients during the first month after liver transplantation. Preoperatively, 5 of the 20 patients had elevated serum levels of parathyroid hormone, and 9 others had low levels of 1,25-(OH)2 vitamin D. After liver transplantation, ectopic calcification was found in seven patients (47%). The organs affected in order of frequency were lungs, liver graft, colon, vascular walls, kidneys, adrenal glands and gastric mucosa. One patient with ectopic calcification of both lungs had markedly restricted pulmonary function as well as radiologic evidence of osteopenia and pathologic fractures of three vertebrae. Postoperatively, increased parathyroid hormone levels were found in all patients who developed soft tissue calcification. Parathyroid hormone levels peaked during the second week after transplantation and were higher at all times compared to subjects without calcification. Five of the seven patients with ectopic calcification had associated renal failure. Individuals who developed calcification had received significantly more fresh frozen plasma, red blood cells and elemental calcium postoperatively, but showed no difference in serum levels of calcium, magnesium, vitamin D, total plasma CO2 or phosphate levels when compared to patients without calcification.(ABSTRACT TRUNCATED AT 250 WORDS)


Nedaplatin and 5-FU combined with radiation in the treatment for esophageal cancer.
A series of patients with esophageal cancer was treated with chemotherapeutic regimens of the new antitumor platinum preparation nedaplatin plus 5-FU in combination with radiation therapy, and the therapeutic responses, side effects, and complications were clinically assessed. There were 2 patients with a complete response and 11 patients with a partial response, hence, a response rate of 76.5%. Major adverse reactions were those of hematological toxicity and included leukopenia (13 patients, 76.5%), thrombocytopenia (8 patients, 47.1%), and lowered serum hemoglobin concentration (9 patients, 52.9%). The leukopenia and thrombocytopenia, though of a grade 3 severity in 3 and 2 patients, respectively, subsided spontaneously in all affected cases. Gastrointestinal adverse reactions were mild and included appetite loss in 7 patients (41.2%) and nausea in 2 patients (11.8%). The only abnormality in renal function observed was a slight elevation of serum creatinine in one patient. The combined therapy of chemotherapy with nedaplatin and 5-FU plus radiation produced a high response rate in the treatment of carcinoma of the esophagus and was associated with reduced gastrointestinal and renal toxicity. The results indicate the combined therapy with nedaplatin to be clinically useful.


Environment and coordination of FeMo-co in the nitrogenase metallochaperone NafY.
In nitrogenase biosynthesis, the iron-molybdenum cofactor (FeMo-co) is externally assembled at scaffold proteins and delivered to the NifDK nitrogenase component by the NafY metallochaperone. Here we have used nuclear magnetic resonance, molecular dynamics, and functional analysis to elucidate the environment and coordination of FeMo-co in NafY. H121 stands as the key FeMo-co ligand. Regions near FeMo-co diverge from H121 and include the η1, α1, α2 helical lobe and a narrow path between H121 and C196.


Teacher attitudes toward educational and psychological services for conduct problem children.
Ninety-nine teachers participated in a survey designed to assess teacher attitudes toward and experience with a variety of strategies for dealing with conduct problem children in regular classes. Results indicated that (a) a majority of teachers favored placing conduct problem children in regular rather than special classes; (b) current sources of assistance (school psychologists, clinical psychologists, principals, parents, and other teachers) were uniformly perceived as only slightly affective in helping teachers handle problem children; and (c) of 14 alternative educational and psychological assistance delivery strategies, teachers most frequently preferred smaller classes, teacher aides, increased counseling staff, and resource classrooms.


Heat Shock Protein 27 is down-regulated in Ballooned Hepatocytes of Patients with Nonalcoholic Steatohepatitis (NASH).
Ballooning degeneration (BD) of hepatocytes is a distinguishing histological feature associated with the progression of nonalcoholic fatty liver disease (NAFLD). Under the assumption that NAFLD severity is associated with metabolic-stress we explored the hypothesis that heat shock 27 kDa protein 1 (HSP27), a protein chaperone involved in stress resistance and cytoskeletal-remodeling, might be deregulated in ballooned hepatocytes. We observed that fasting plasma glucose (fpG) (p = 0.00002), total cholesterol (p = 0.02) and triglycerides (p = 0.01) levels, and female sex (p = 0.01) were significantly associated with the presence of BD. A logistic regression model showed that BD was independently associated with fpG (p = 0.002); OR per unit of glucose concentration 1.05, 95% confidence interval 1.02-1.09. Furthermore, BD was associated with a significant 2.24-fold decrease in the expression level of HSP27-mRNA in comparison with absence of ballooning, p = 0.002. Ballooned hepatocytes showed very low HSP27 immunoreactivity compared with hepatocyes without ballooning (p = 0.009); HSP27 immunoreactivity was inversely correlated with fpG levels (R: -0.49, p = 0.01). In conclusion, BD is associated with down-regulation of liver HSP27 gene and protein expression, suggesting that ballooned hepatocytes fail to ensure a robust physiological response to metabolic-induced stress.


The subunit composition of factor XIII proteins in normal and factor XIII deficient plasma and serum analysed by line immunoelectrophoresis.
Line immunoelectrophoresis was used for the characterization of human coagulation factor XIII in normal and factor XIII-deficient plasma and serum. In normal plasma, two immunoreactive factor XIII proteins termed XIII-p (Mr 330,000 Da) and XIII-s (Mr 170,000 Da) were identified within the beta-globulin fraction. Their reactions with monospecific antisera against known factor XIII subunits, suggests that the factor XIII-p and -s proteins recognized represent the factor XIII a2b2 tetramer and -b2 dimer, respectively. The concentration of the XIII-p protein was, in normal plasma, approximately 6 mg/l, in plasma from a case of congenital factor XIII, approximately 1 mg/l and in normal serum approximately 3 mg/l. The concentration of the XIII-s protein (approximately 18 mg/l) was unaffected by the coagulation process and normal in plasma and serum from the case of congenital factor XIII deficiency. Thus, the present investigation indicates the presence of an excess of the anticatalytic, DNA-binding factor XIII carrier protein in normal as well as in factor XIII-deficient plasma and serum.


[Disorders in the consolidation of temporary connections in the rat after chemical induction of lipid peroxidation and its prevention with the antioxidant ionol].
Injections to rats of antibiotic adriblastin activating lipids peroxide oxidation directly after elaboration of defensive conditioned reflexes, elicit retrograde amnesia. Administration of antioxidant ional before the administration of adriblastin prevents this effect. It is suggested that activation of lipids peroxide oxidation disturbs the process of consolidation.


Soluble guanylate cyclase stimulator riociguat improves spatial memory in mice via peripheral mechanisms.
Soluble guanylate cyclase (sGC) - cyclic guanosine monophosphate (cGMP) signalling is important for healthy memory function and a healthy vascular system. Targeting sGC-cGMP signalling can therefore be a potential strategy to enhance memory processes. sGC can be targeted by using agonists, such as sGC stimulator riociguat. Therefore, this study aimed to target sGC using riociguat to investigate its acute effects on memory function and neuronal plasticity in mice. The effects of riociguat on long-term memory and a biperiden-induced memory deficit model for assessing short-term memory were tested in the object location task, and working memory was tested in the Y-maze continuous alternation task. Pharmacokinetic measurements were performed within brain tissue of mice, and hippocampal plasticity measures were assessed using western blotting. Acute oral administration with a low dose of 0.03 mg/kg riociguat was able to enhance working-, short-, and long-term spatial memory. Under cerebral vasoconstriction higher doses of riociguat were still effective on memory. Pharmacokinetic measurements revealed poor brain penetration of riociguat and its metabolite M-1. Increased activation of VASP was found, while no effects were found on other memory-related hippocampal plasticity measures. Memory enhancing effects of riociguat are most likely regulated by vascular peripheral effects on cGMP signalling. Yet, further research is needed to investigate the possible contribution of hemodynamic or metabolic effects of sGC stimulators on memory performance.


An iron-rich organelle in the cuticular plate of avian hair cells.
Hair cells reside in specialized epithelia in the inner ear of vertebrates, mediating the detection of sound, motion, and gravity. The transduction of these stimuli into a neuronal impulse requires the deflection of stereocilia, which are stabilized by the actin-rich cuticular plate. Recent electrophysiological studies have implicated the vestibular system in pigeon magnetosensation. Here we report the discovery of a single iron-rich organelle that resides in the cuticular plate of cochlear and vestibular hair cells in the pigeon. Transmission electron microscopy, coupled with elemental analysis, has shown that this structure is composed of ferritin-like granules, is approximately 300-600 nm in diameter, is spherical, and in some instances is membrane-bound and/or organized in a paracrystalline array. This organelle is found in hair cells in a wide variety of avian species, but not in rodents or in humans. This structure may function as (1) a store of excess iron, (2) a stabilizer of stereocilia, or (3) a mediator of magnetic detection. Given the specific subcellular location, elemental composition, and evolutionary conservation, we propose that this structure is an integral component of the sensory apparatus in birds.


Species characteristics of serum proteins demonstrated after agarose gel electrophoresis.
Serum samples from five species of domestic animals were subjected to protein electrophoresis on commercially prepared agarose gel films. After processing, the strained electrophoretic patterns and densitometric traces exhibited certain characteristics for each species. The features which were considered distinctive of each species are illustrated.


Asymmetric Construction of Functionalized 1,2-Dihydropyridine and Pyridine Derivatives with Adjacent Stereocenters via a Unified Metal-Free Catalytic Approach.
A novel asymmetric catalytic approach for the construction of enantioenriched functionalized 1,2-dihydropyridines and pyridine derivatives incorporating adjacent quaternary and tertiary stereocenters has been reported. This process involved a metal-free catalytic asymmetric allylic alkylation and a stereospecifically nonoxidative aromatization approach for the desired chiral molecules.


Preclinical evaluation of gene transfer products: safety and immunological aspects.
As any new drug in development, gene transfer products have to be tested for their potential toxicity in preclinical studies. Strategies for preclinical safety evaluation of gene transfer compounds are similar to those undertaken for biotechnology-derived pharmaceuticals with the added complexity of having to test additional components, such as the vector and genetic material. Some recommendations have been issued by regulatory agencies to provide assistance in the toxicological assessment of gene transfer products before administration to humans. However, the design of such studies is complicated and has to be approached case-by-case. Crucial aspects for safety evaluation are: (1) the choice of animal models with respect to the pharmacological activity of the gene transfer product, the susceptibility to infection in the case of viral vectors, and to the immunogenicity of all entities; (2) biodistribution of the vector after administration, avoiding germinal transmission. Immunological and immunotoxicity endpoints should also be assessed. This paper reviews some of the properties of gene transfer vectors with emphasis on their potential effects on the immune system, and also highlights some of the problems likely to be encountered during the preclinical safety evaluation.


How an overlooked gene in coenzyme a synthesis solved an enzyme mechanism predicament.
Coenzyme A (CoA) is an essential cofactor throughout biology. The first committed step in the CoA synthetic pathway is synthesis of β-alanine from aspartate. In Escherichia coli and Salmonella enterica panD encodes the responsible enzyme, aspartate-1-decarboxylase, as a proenzyme. To become active, the E. coli and S. enterica PanD proenzymes must undergo an autocatalytic cleavage to form the pyruvyl cofactor that catalyzes decarboxylation. A problem was that the autocatalytic cleavage was too slow to support growth. A long-neglected gene (now called panZ) was belatedly found to encode the protein that increases autocatalytic cleavage of the PanD proenzyme to a physiologically relevant rate. PanZ must bind CoA or acetyl-CoA to interact with the PanD proenzyme and accelerate cleavage. The CoA/acetyl-CoA dependence has led to proposals that the PanD-PanZ CoA/acetyl-CoA interaction regulates CoA synthesis. Unfortunately, regulation of β-alanine synthesis is very weak or absent. However, the PanD-PanZ interaction provides an explanation for the toxicity of the CoA anti-metabolite, N5-pentyl pantothenamide.


Muyin extract inhibits non-small-cell lung cancer growth by inducing autophagy and apoptosis in vitro and in vivo.
Non-small cell lung cancer (NSCLC) is a major subtype of lung cancer with a higher mortality rate. Both apoptosis and autophagy are crucial processes in the pathophysiology of NSCLC. Muyin extract (MSE) is a combination of Momordica cochinchinensis (Lour.) Spreng seeds and Epimedium brevicornu Maxim extract, with an optimal ratio of 1:1. Our previous research has firstly shown that MSE exerts a good anti-tumor activity, especially for NSCLC. This study aims to evaluate the inhibitory effect of MSE on NSCLC and explore the underlying mechanism. In vitro, cell proliferation was examined by MTT and colony formation. Apoptosis was detected by annexin V-FITC/PI assay while autophagy was assessed by Acridine orange (AO) and Monodansylcadaverine (MDC) staining. In vivo, Lewis lung cancer cell transplanted mice model was established to measure the effect of MSE on tumor growth. Hematoxylin eosin (H & E) staining was used to observe the pathological changes of the tumor after MSE treatment. The apoptosis in tumor tissue was detected by TUNEL assay. Meanwhile, the cellular proliferation marker Ki67 and autophagy marker LC3Ⅱ were observed by immunohistochemistry staining. The IL-4 and IFN-γ concentrations in blood were tested by Elisa. The apoptosis related factors (Bcl-2, Bax Caspase-3, cleaved Caspase-3, Caspase-9 and p53), autophagy marker proteins (Atg-5, Becline-1, LC3Ⅱ/Ⅰand p62) as well as Akt/mTOR pathway were detected by western blotting. Present study showed that MSE greatly inhibited the proliferation of NSCLC in vitro and in vivo, together with apoptotic rate increasing. P53 and cleaved Caspase-3 levels were up-regulated while Bcl-2/Bax ratio, Caspase-3 and Caspase-9 levels were significantly down-regulated treated with MSE. Meanwhile, MSE activated autophagy, Atg-5, Becline-1 as well as the ratio of LC3Ⅱ/Ⅰ were notably up-regulated while p62 was down-regulated after MSE treatment. Importantly, MSE significantly blocked Akt/mTOR pathway, which is a common upstream signal triggered by autophagy and apoptosis. Furthermore, when co-treated with specific autophagy inhibitor, the inhibitory rate and anti-apoptotic Bcl-2 level were significantly reversed. Impressively, MSE remarkably increased IFN-γ/ IL-4 ratio while VP16 did not in animal model, and the inhibition rate in tumor weight after MSE treatment was higher than xiaojin pill. Taken together, it is proved that MSE may be a promising oral TCM candidate for NSCLC therapy with immunity improvement. The underlying mechanisms could be associated with the induction of apoptosis and autophagy through blocking Akt/mTOR pathway, meanwhile, it may promote crosstalk between autophagy and apoptosis.


Effects of normobaric oxygen on the progression of focal cerebral ischemia in rats.
Normobaric oxygen (NBO) reduces infarction at 24-48 h in experimental models of focal cerebral ischemia. However, to be clinically relevant, longer term safety and efficacy must be explored. Here, we assessed the effects of NBO on glial activation, neurovascular recovery, and behavioral outcomes at 2 weeks after transient focal ischemia in rats. 100 min transient focal ischemia was induced by intraluminal occlusion of the middle cerebral artery in adult male Sprague-Dawley rats. Animals were randomized into sham, controls or 85'NBO started 15 min after ischemic onset. Infarct volumes and behavioral outcomes were blindly quantified. Immunohistochemistry was used to examine the effects of NBO on glial activation and neurovascular responses. After 2 weeks of reperfusion the infarct volume was marked reduced in animals subjected to NBO. They also had better outcomes in forelimb placement test and in body-swing test and weight loss reduction. After 14 days, NBO decreased expression of Iba1, a marker of activated microglia, and GFAP, a marker of activated astrocytes. NBO treatment had no detectable effect on angiogenesis. These results suggest that protective effects of NBO may persist for up to 2 weeks post-stroke.


CIU and main event analyses of the structured discourse of older and younger adults.
Correct information unit (CIU) and main event analyses are quantitative measures for analyzing discourse of individuals with aphasia. Comparative data from healthy younger (YG) and older (OD) adults and an investigation of the influence of stimuli type would considerably extend the usefulness of such analyses. The objectives were (a) to compare discourse ability of younger and older healthy adults, and (b) to investigate influence of stimuli type on performance. Seventeen younger and 17 older healthy adults participated. Language samples obtained were analyzed for CIU and proportion of main events. Results indicated the YG group produced more accurate, relevant, and informative content than the OD group, and participants expressed more main events in response to sequential picture stimuli, regardless of age. We conclude that clinicians should consider stimuli type when measuring discourse ability. Age should also be considered when comparing performance of adults with brain damage to that of healthy adults. The reader will be able to: (a) recognize the importance of obtaining information about discourse performance in both older and younger healthy adults; (b) identify the purposes of CIU analysis and main event analysis for examining discourse; and (c) appreciate the potential influences of different language elicitation procedures on the discourse obtained from a task.


Investigations on the anti-inflammatory and anti-allergic activities of the leaves of Uncaria guianensis (Aublet) J. F. Gmelin.
Uncaria guianensis (Aublet) J. F. Gmelin is an herbal medicine from tropical areas of South and Central America. We investigated the anti-inflammatory and anti-allergic properties of an ethanolic extract of U. guianensis leaves, containing alkaloids, flavonoids and phenol carboxylic acids, as revealed by thin layer chromatography (TLC). Oral pre-treatment with U. guianensis inhibited zymosan-induced paw oedema (500 mg/paw) and pleural exudation (100 mg/kg) within 4 h (25-200 mg/kg). U. guianensis (100 mg/kg) inhibited total leukocyte and neutrophil numbers in the pleural cavity 4 h after zymosan stimulation. Pre-treatment with U. guianensis (100 mg/kg, p.o.) inhibited total leukocyte, neutrophil and eosinophil recruitment into the pleural cavity 24 h after LPS (250 ng/cavity, i.t.). Pre-treatment with U. guianensis inhibited paw oedema (25-200 mg/kg) induced by ovalbumin (OVA) within 1 h, and neutrophil and eosinophil recruitment into the mice pleural cavity 24 h after OVA (100 mg/kg). In vitro data revealed that U. guianensis impaired LPS-induced nitric oxide and CXCL8 generation by murine peritoneal macrophages, as well as OVA-induced interleukin-5 synthesis by previously sensitized spleen cells. These results demonstrate that U. guianensis leaves provide effective anti-inflammatory and anti-allergic activities.


Outcomes After Stage I Norwood Palliation in a Recently Established Program Can Achieve Results Similar to Longer Established Services.
Infants with hypoplastic left heart syndrome (HLHS) or similar single ventricle cardiac lesions require a three-stage surgical approach, the first step being the Stage I Norwood procedure. The Queensland Children's Hospital (QCH) in Australia is a tertiary hospital providing the only cardiac surgical service to children in Queensland and northern New South Wales. To review the centre's outcomes of Norwood procedures performed in the last 6 years. We retrospectively evaluated all infants undergoing the stage I Norwood procedure between January 2015 and August 2021. Mortality, intensive care length of stay, events of cardiac arrest following surgery and duration of mechanical ventilation were calculated and analysed for subgroups depending on type of pulmonary shunt type (right-ventricle-to-pulmonary-artery shunt [RVPAS] vs the modified Blalock-Taussig shunt [MBTS]). Forty-nine (49) patients were included. Overall survival to stage two operation (Glenn) was 90%. Both shunts were used evenly with the RVPA conduit preferred for HLHS and the MBTS largely chosen for hypoplastic left heart variants. In univariable analysis there was no difference in cardiac arrest or mortality rate for the patient with a RVPAS compared to the patient with a MBTS. We show that a recently established Norwood program can achieve results that are comparable to those reported by longer established centres, and the international literature.


Geometric and dosimetric accuracy and imaging dose of the real-time tumour tracking system of a gimbal mounted linac.
To suggest a comprehensive testing scheme to evaluate the geometric and dosimetric accuracy and the imaging dose of the VERO dynamic tumour tracking (DTT) for its clinical implementation. Geometric accuracy was evaluated for gantry 0° and 90° in terms of prediction (EP), mechanical (EM) and tracking (ET) errors for sinusoidal patterns with 10 and 20 mm amplitudes, 2-6 s periods and phase shift up to 1 s and for 3 patient patterns. The automatic 4D model update was investigated simulating changes in the breathing pattern during treatment. Dosimetric accuracy was evaluated with gafchromic films irradiated in static and moving phantom with and without DTT. The entrance skin dose (ESD) was assessed using a solid state detector and gafchromic films. The RMS of EP, EM, and ET were up to 0.8, 0.5 and 0.9 mm for all non phased-shifted motion patterns while for the phased-shifted ones, EP and ET increased to 2.2 and 2.6 mm. Up to 4 updates are necessary to restore a good correlation model, according to type of change. For 100 kVp and 1 mA s X-ray beam, the ESD per portal due to 20 s fluoroscopy was 16.6 mGy, while treatment verification at a frequency of 1 Hz contributed with 4.2 mGy/min. The proposed testing scheme highlighted that the VERO DTT system tracks a moving target with high accuracy. The automatic update of the 4D model is a powerful tool to guarantee the accuracy of tracking without increasing the imaging dose.


Lambda phage-based vaccine induces antitumor immunity in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a difficult to treat tumor with a poor prognosis. Aspartate β-hydroxylase (ASPH) is a highly conserved enzyme overexpressed on the cell surface of both murine and human HCC cells. We evaluated therapeutic effects of nanoparticle lambda (λ) phage vaccine constructs against ASPH expressing murine liver tumors. Mice were immunized before and after subcutaneous implantation of a syngeneic BNL HCC cell line. Antitumor actively was assessed by generation of antigen specific cellular immune responses and the identification of tumor infiltrating lymphocytes. Prophylactic and therapeutic immunization significantly delayed HCC growth and progression. ASPH-antigen specific CD4+ and CD8+ lymphocytes were identified in the spleen of tumor bearing mice and cytotoxicity was directed against ASPH expressing BNL HCC cells. Furthermore, vaccination generated antigen specific Th1 and Th2 cytokine secretion by immune cells. There was widespread necrosis with infiltration of CD3+ and CD8+ T cells in HCC tumors of λ phage vaccinated mice compared to controls. Moreover, further confirmation of anti-tumor effects on ASPH expressing tumor cell growth were obtained in another murine syngeneic vaccine model with pulmonary metastases. These observations suggest that ASPH may serve as a highly antigenic target for immunotherapy.


[Inflammatory diseases : anatomo-pathological features].
The aim of this article is to describe the role of anatomic pathology in the management of acute and chronic inflammatory diseases. Granulomatous pathologies will be also addressed as well as some dysimmune pathologies and recent data on the prognostic role of the antitumoral inflammatory response.


Evidence for the nonenzymatic and irreversible binding of cytembena to rat liver microsomes in vitro.
Characteristics of the irreversible binding of cytembena (CYT) to rat liver microsomal proteins have been investigated in vitro. Binding of [14C]CYT to rat liver microsomal proteins remained unchanged in the presence of SKF-525A or after heat denaturation and was not dependent upon the presence of pyridine nucleotide (NADPH or NADH). Ethacrynic acid, cysteine, dithiothreitol, and lysine were found to block the irreversible binding of [14C]CYT to microsomal proteins in a dose-dependent manner. The rank order of effectiveness as inhibitors of CYT binding was ethacrynic acid greater than cysteine greater than dithiothreitol greater than lysine. In other studies, CYT was shown to preferentially form adducts with cysteine rather than lysine or glycine. Using structural analogs and metabolites of CYT, it was found that 4-methoxybenzoylacrylic acid and beta-benzoylacrylic acid were effective competitors of [14C]CYT binding to liver microsomal proteins. By contrast, 4-methoxybenzoylpropionic acid, 5-(4-methoxyphenyl)dihydro-2(3H)furanone and 4-hydroxy-4-(4-methoxyphenyl)butyric acid were ineffective as inhibitors of the irreversible binding of CYT. These data suggest that sulfhydryl groups are involved in the nonenzymatic binding of CYT and that the presence of a carbon-carbon double bond in CYT, in debrominated metabolites or structural analogs, is requisite for an interference of the binding of CYT to microsomal proteins.


Identification of type II interferons and receptors in an osteoglossiform fish, the arapaima Arapaima gigas.
In mammals, type II interferon (IFN; i.e. IFN-γ) signalling transduces through its specific receptors IFN-γR1 and IFN-γR2. In an osteoglossiform fish, the arapaima Arapaima gigas, three type II IFNs, IFN-γ-like, IFN-γ and IFN-γrel, and their four possible receptor subunits IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 were identified in this study. The three type II IFN genes are composed of four exons and three introns, and they all contain IFN-γ signature motif and signal peptide, with the presence of potential nuclear localization signal (NLS) in IFN-γ-like and IFN-γ. The IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 are composed of seven exons and six introns, with predicted IFN-γR1-1 and IFN-γR1-2 proteins containing JAK1 and STAT1 binding sites, and IFN-γR2-1 and IFN-γR2-2 containing JAK2 binding sites. Gene synteny analysis showed that the type II IFN and their receptor loci are duplicated in arapaima. All these genes were expressed constitutively in all organs/tissues examined, and responded to the stimulation of polyI:C. The prokaryotic recombinant IFN-γ-like, IFN-γ and IFN-γrel proteins can significantly induce the upregulation of immune-related genes in trunk kidney leucocytes. The ligand-receptor relationship analyses revealed that recombinant IFN-γ-like, IFN-γ, and IFN-γrel transduce downstream signalling through IFN-γR1-1/IFN-γR2-1, IFN-γR1-2/IFN-γR2-2, and IFN-γR1-1, respectively, in xenogeneic cells with the overexpression of original or chimeric receptors. In addition, tyrosine (Y) 366 and Y377 in the intracellular region may be essential for the function of IFN-γR1-2 and IFN-γR1-1, respectively. The finding of type II IFN system in A. gigas thus provides different knowledge in understanding the diversity and evolution of type II IFN ligand-receptor relationships in vertebrates.


Relevance of using length of stay as a key indicator to monitor emergency department performance: Case study from a rural hospital in Thailand.
The present study explores factors related to length of stay (LOS) in a rural public hospital in Thailand and assesses the feasibility of using LOS as an ED key performance indicator. Using a mixed-methods approach, qualitative methods (in-depth interviews, patients' chart review and participatory observations) were used to guide and elaborate findings from quantitative analysis of 555 electronic ED records. Multivariate analysis revealed that age, Emergency Severity Index score and number of laboratory tests were significantly associated with LOS. The qualitative approach provided contradicting evidence on the linkage between LOS and patient outcomes. On the one hand, considering the 4 h rule, a child with asthma was referred to a tertiary care hospital because of deterioration after 4 h of ED care. On the other hand, a woman with sepsis was hospitalised with improved condition despite 7 h of ED care. Interviews revealed the waiting time to see doctors was probably the top priority issue for patients. Factors related to LOS in a rural hospital in Thailand are similar and in contrast to those of a previous study in a medical school setting. Reasons for the discrepancy of findings and implications for improving ED services were discussed. Our data support the notion of controversy in using LOS as a key indicator of ED performance in this rural hospital setting. Thus, it is imperative to not rely on any single throughput or process indicators to monitor ED performance, but to take into account a set of indicators including patient outcomes.


Activity of multipurpose contact lens solutions against Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens and Candida albicans biofilms.
The use of contact lenses has progressively increased around the world, thereby increasing the risk of complications. The most serious complication is microbial keratitis (corneal infection) that can progress to a corneal ulcer. Fourteen multipurpose contact lens solutions were tested on mature biofilms comprising Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens and Candida albicans, using the minimum disinfection times recommended by the manufacturers. The biofilm was induced in the lens case, and 24 h later, the solutions were added. Activity against planktonic and sessile cells was evaluated and quantified as colony forming units per millilitre. The minimum concentration for biofilm eradication was defined as a 99.9% reduction in viable cells. Although most solutions exhibited activity against planktonic cells, only five of the 14 solutions produced a significant reduction in the S. marcescens biofilm. No solution achieved the minimal biofilm eradication of S. aureus, P. aeruginosa and C. albicans. Multipurpose contact lens solutions provide greater bactericidal and/or fungicidal activity on planktonic cells than biofilms. The minimal eradication biofilm concentration was only achieved for S. marcescens.


Results from the Effects of MEtformin on cardiovasculaR function in AdoLescents with type 1 Diabetes (EMERALD) study: A brief report of kidney and inflammatory outcomes.
Youth with type 1 diabetes (T1D) demonstrate insulin resistance, independently of glycaemia, when compared to normoglycaemic peers. Insulin resistance increases the risk of cardiovascular disease and diabetic kidney disease, factors also associated with systemic inflammation. We evaluated the effect of metformin on markers of inflammation and diabetic kidney disease in adolescents with T1D. EMERALD, a double-blind, randomized, placebo-controlled trial of 3 months of metformin in 48 participants aged 12-21 years with T1D, included baseline and follow-up assessments of serum creatinine and cystatin C to estimate glomerular filtration rate (eGFR), aspartate aminotransferase, alanine aminotransferase, high-sensitivity C-reactive protein, white blood count, platelets, adiponectin, leptin, and urine albumin: creatinine ratio (UACR). Metformin was associated with a 13.9 mL/min/1.73 m2 (95% confidence interval 4.7-23.1 mL/min/1.73 m2 ) increase in estimated GFR by serum creatinine versus placebo (P ≤ 0.01), with a significant difference remaining after multivariable adjustments (P = 0.03). Whereas eGFR measured by serum creatinine increased significantly after metformin treatment, no differences were observed in cystatin C, UACR, or systemic inflammatory markers. Additional studies with directly measured GFR in response to metformin in T1D are needed.


Defined microbiota transplant restores Th17/RORγt+ regulatory T cell balance in mice colonized with inflammatory bowel disease microbiotas.
The building evidence for the contribution of microbiota to human disease has spurred an effort to develop therapies that target the gut microbiota. This is particularly evident in inflammatory bowel diseases (IBDs), where clinical trials of fecal microbiota transplantation have shown some efficacy. To aid the development of novel microbiota-targeted therapies and to better understand the biology underpinning such treatments, we have used gnotobiotic mice to model microbiota manipulations in the context of microbiotas from humans with inflammatory bowel disease. Mice colonized with IBD donor-derived microbiotas exhibit a stereotypical set of phenotypes, characterized by abundant mucosal Th17 cells, a deficit in the tolerogenic RORγt+ regulatory T (Treg) cell subset, and susceptibility to disease in colitis models. Transplanting healthy donor-derived microbiotas into mice colonized with human IBD microbiotas led to induction of RORγt+ Treg cells, which was associated with an increase in the density of the microbiotas following transplant. Microbiota transplant reduced gut Th17 cells in mice colonized with a microbiota from a donor with Crohn's disease. By culturing strains from this microbiota and screening them in vivo, we identified a specific strain that potently induces Th17 cells. Microbiota transplants reduced the relative abundance of this strain in the gut microbiota, which was correlated with a reduction in Th17 cells and protection from colitis.


Aberrant subclavian arteries: cross-sectional imaging findings in infants and children referred for evaluation of extrinsic airway compression.
The purpose of our study was to describe patterns of airway compression identified on cross-sectional imaging in infants and children with either right aortic arch and aberrant left subclavian artery or left aortic arch with aberrant right subclavian artery. Data from MR imaging and CT performed to evaluate pediatric patients for extrinsic airway compression were reviewed for cases that revealed an aberrant right or left subclavian artery. Clinical, endoscopic, and imaging findings in identified cases were reviewed. Recurrent patterns of extrinsic compression were reviewed among cases. Twelve patients with right aortic arch with aberrant left subclavian artery and nine patients with left aortic arch and aberrant right subclavian artery were identified. All 12 with right aortic arch with aberrant left subclavian artery had airway compression shown, with multiple sites or diffuse compression in six. Of these 12 patients, nine had compression at the level of the arch and aberrant subclavian artery (10 had Kommerell's diverticulum), and nine had compression of the distal airway in association with a midline descending aorta. Five of the nine patients with left aortic arch and aberrant right subclavian artery had airway compression shown, all at the level of the arch and aberrant subclavian artery. None of these compressions was associated with either Kommerell's diverticulum or midline descending aorta. Both right and left aberrant subclavian arteries can be associated with symptomatic airway compression, but the patterns of compression are different. The airway compression in right aortic arch with aberrant left subclavian artery is often associated with either Kommerell's diverticulum or midline descending aorta, whereas compression associated with left aortic arch and aberrant right subclavian artery is not.


Recommendations for the treatment of Sjögren's syndrome.
The recommendations proposed by the Sjögren's Syndrome Committee of the Brazilian Society of Rheumatology for the treatment of Sjögren's syndrome were based on a systematic review of literature in Medline (PubMed) and the Cochrane databases until October 2014 and on expert opinion in the absence of studies on the subject. 131 items classified according to Oxford & Grade were included. These recommendations were developed in order to guide the appropriate management and facilitate the access to treatment for those patients with an appropriate indication, considering the Brazilian socioeconomic context and pharmacological agents available in this country.


Prevalence of symptoms and use of medication for gastroesophageal reflux in an Australian community.
Gastroesophageal reflux disease is common in Western societies, although the prevalence of reflux symptoms in the community is not well described. In this study we determined the prevalence of symptoms of gastroesophageal reflux and other "esophageal" symptoms, and the consumption of medication for reflux in an Australian community. A population sample designed to accurately reflect the characteristics of the population aged 15 years or older in the State of South Australia was studied. Demographic data; symptoms specific to reflux, dysphagia, and abdominal bloating; and the consumption of antireflux medication were determined in a face-to-face interview. The frequency and severity of heartburn and dysphagia were assessed with analog scales. A total of 2,973 people (age range: 15-95 years) were interviewed between September and December 2006. Approximately half experienced the symptom of heartburn; 21.2% experienced heartburn at least once a month, and 12.4% described frequent symptoms of heartburn (at least a few times each week). Of those with heartburn, 25.0% graded it as moderate or severe, 10.9% reported some dysphagia for solid foods, and 6.9% reported dysphagia for liquids. 3.7% described dysphagia for solids at least once a month. Abdominal bloating was reported by 48.2%. 16.9% were taking medications for reflux symptoms (10.1% proton pump inhibitors, 1.2% H2-receptor antagonists, 2.1% simple antacids, 3.4% alternative medications). Heartburn was more common in individuals who consumed medication. There were significant associations between heartburn and bloating, and between heartburn and dysphagia. Symptoms of gastroesophageal reflux and the use of medications to treat such symptoms are very common in the community of South Australia. Nearly 1 in 7 people over the age of 15 consume medication for the treatment of symptoms of reflux.


The impact of switching P2Y12 receptor inhibitor therapy during index hospitalization: a systematic review.
With the availability of novel P2Y12 receptor inhibitors, patients presenting with acute coronary syndrome (ACS) may receive more than one type of this drug during index hospitalization. We sought to determine the effect of switching from clopidogrel to a novel P2Y12 receptor inhibitor on the occurrence of major adverse cardiovascular events (MACE) and bleeding. We conducted a literature search on SCOPUS for English language entries until 7 March 2015. Out of 188 citations, seven studies encompassing 16,431 patients were selected for analysis of (i) switching to a novel P2Y12 agent (switching group) versus continued clopidogrel or (ii) switching to a novel P2Y12 agent (switching group) versus upfront novel agent initiation during index hospitalization MACE was significantly lower in the switching group (odds ratio (OR) 0.77, 95 % confidence interval (CI) 0.63-0.96, p = 0.02), whereas bleeding was higher (OR 1.55, 1.29-1.85, p < 0.01) compared with continued clopidogrel. Conversely, MACE was similar with switching to a novel agent and upfront novel therapy initiation (OR 1.01, 95 % CI 0.8-1.29, p = 0.90), but bleeding was higher in the switching group (OR 1.24, 95 % CI 1.03-1.48, p = 0.02). The current study suggests that switching to a novel P2Y12 agent in patients with ACS and/or patients undergoing coronary stenting is more efficacious than continuing clopidogrel. In this cohort, switching to a novel agent did not result in worse ischemic outcomes than upfront initiation of novel therapies. However, switching was associated with greater bleeding compared with both continued clopidogrel as well as upfront use of novel P2Y12 agents.


Gd DTPA. Clinical efficacy.
Both the diagnostic accuracy and sensitivity of the MRI diagnosis of central nervous system neoplasms appear to be improved by the administration of a paramagnetic contrast agent, Gd DTPA.


The RNA polymerase III-RIG-I axis in antiviral immunity and inflammation.
Nucleic acid sensors survey subcellular compartments for atypical or mislocalized RNA or DNA, ultimately triggering innate immune responses. Retinoic acid-inducible gene-I (RIG-I) is part of the family of cytoplasmic RNA receptors that can detect viruses. A growing literature demonstrates that mammalian RNA polymerase III (Pol III) transcribes certain viral or cellular DNA sequences into immunostimulatory RIG-I ligands, which elicits antiviral or inflammatory responses. Dysregulation of the Pol III-RIG-I sensing axis can lead to human diseases including severe viral infection outcomes, autoimmunity, and tumor progression. Here, we summarize the newly emerging role of viral and host-derived Pol III transcripts in immunity and also highlight recent advances in understanding how mammalian cells prevent unwanted immune activation by these RNAs to maintain homeostasis.


Plasma Treatment Limits Human Melanoma Spheroid Growth and Metastasis Independent of the Ambient Gas Composition.
Melanoma skin cancer is still a deadly disease despite recent advances in therapy. Previous studies have suggested medical plasma technology as a promising modality for melanoma treatment. However, the efficacy of plasmas operated under different ambient air conditions and the comparison of direct and indirect plasma treatments are mostly unexplored for this tumor entity. Moreover, exactly how plasma treatment affects melanoma metastasis has still not been explained. Using 3D tumor spheroid models and high-content imaging technology, we addressed these questions by utilizing one metastatic and one non-metastatic human melanoma cell line targeted with an argon plasma jet. Plasma treatment was toxic in both cell lines. Modulating the oxygen and nitrogen ambient air composition (100/0, 75/25, 50/50, 25/75, and 0/100) gave similar toxicity and reduced the spheroid growth for all conditions. This was the case for both direct and indirect treatments, with the former showing a treatment time-dependent response while the latter resulted in cytotoxicity with the longest treatment time investigated. Live-cell imaging of in-gel cultured spheroids indicated that plasma treatment did not enhance metastasis, and flow cytometry showed a significant modulation of S100A4 but not in any of the five other metastasis-related markers (β-catenin, E-cadherin, LEF1, SLUG, and ZEB1) investigated.


Effect of chemical structure of interface modifier of TiO2 on photovoltaic properties of poly(3-hexylthiophene)/TiO2 layered solar cells.
Two classes of phosphonic acid-bearing organic molecules, 2-oligothiophene phosphonic acid and omega-(2-thienyl)alkyl phosphonic acid were adopted as interface modifiers (IMs) of the TiO(2) surface, to increase its compatibility with poly(3-hexylthiophene) (P3HT). The self-assembled monolayers of these molecules on titania surface were characterized by making contact angle measurements and X-ray photoelectron spectroscopy (XPS). Atomic force microscopic (AFM) images revealed that the adsorption of IMs effectively smooths the TiO(2) surface. Both photoluminescence (PL) spectroscopy and PL lifetime measurements were made to investigate the photoinduced properties of the TiO(2)/IM/P3HT layered-junction. The PL quenching efficiency increased with the number of thiophene rings and as the alkyl chain-length in IMs decreased. Meanwhile, the decline in the PL lifetime followed a similar trend as the PL quenching efficiency. Additionally, the power conversion efficiency (PCE) of the ITO/TiO(2)/IM/P3HT/Au devices was examined by measuring their photocurrent density-applied voltage (J-V) curves. The experimental results indicated that the short-circuit current density (J(SC)) increased with the number of thiophene units and as the hydrocarbon chain-length in IMs decreased. However, the open-circuit voltage (V(OC)) of the devices slightly fell as the energy level of the highest occupied molecular orbital (HOMO) of IM decreased. The PCE of the device with 2-terthiophene phosphonic acid was 2.5 times that of the device with 10-(2-thienyl)decyl phosphonic acid.


Influence of modest endotoxemia on postoperative antithrombin deficiency and circulating secretory immunoglobulin a levels.
To evaluate the influence of modest endotoxemia on postoperative antithrombin deficiency and cholestasis. It has not been determined whether endotoxin translocation in small amounts is a physiological phenomenon or whether it is a potential health hazard. Blood endotoxin, antithrombin III (ATIII), secretory immunoglobulin A (sIgA), which was selected as a marker of cholestasis, C-reactive protein (CRP), and alpha-1-antitrypsin (AAT) concentrations were measured from the 20 patients undergoing curative gastrectomy for gastric cancer preoperatively and postoperatively. Portal and systemic blood samples were taken for the analysis of endotoxin and interleukin-6 (IL-6) concentrations during surgery in these patients. Although plasma endotoxin levels showed a significant increase during surgery, we did not find a correlation with ATIII, sIgA, CRP, and IL-6 levels. Systemic blood endotoxin levels during surgery correlated with a postoperative rise of serum AAT levels. Plasma ATIII levels transiently decreased on the first and third postoperative day, and sIgA levels were shown to increase on the seventh postoperative day. There was a weak relationship between the extent of postoperative endotoxemia and a reduction in ATIII concentrations. The influence of modest endotoxemia on postoperative antithrombin deficiency and cholestasis was limited, and increased translocational endotoxemia during abdominal surgery may be a physiological phenomenon to trigger off an acute-phase protein response.


The role of alcohol-specific socialization in adolescents' drinking behaviour.
To determine which alcohol-specific socialization practices are related to adolescents' alcohol use, and to investigate whether parents differ in their alcohol-specific socialization towards their children. In a sample of 428 families, both parents and two adolescents (aged 13-16 years) completed a questionnaire at home about alcohol-specific parenting and their own alcohol use. Based on the reports of each family member, three different models of alcohol-specific socialization were formulated: from the perspective of the siblings, the mother and the father. Results of structural equation modelling generally showed the same associations between alcohol-specific socialization and drinking of younger and older adolescents. The strongest association was found for providing alcohol-specific rules. Applying strict rules about alcohol use was negatively related to adolescents' alcohol use; this was also the case for having confidence in the effectiveness of alcohol-specific socialization. Unexpectedly, frequency of communication about alcohol issues was positively associated with alcohol consumption of adolescents. This study is one of the first to examine associations between alcohol-specific socialization and adolescents' drinking using a between- and a within-family design. Results showed strong associations between alcohol-specific socialization (particularly of enforcing rules) and adolescent alcohol use. Although parents strongly differentiated their socialization practices between children, no differences in associations between alcohol-specific socialization and drinking were found between older and younger adolescents.


Systematic review of quality of reporting, outcome measurements and methods to study efficacy of preventive and therapeutic approaches to peri-implant diseases.
To systematically review the literature and to assess the quality of reporting, outcome measurements and methods in both preventive and therapeutic approaches to peri-implant mucositis (PM) and peri-implantitis (PI). Randomized (RCT) and Controlled Clinical Trials (CCT), evaluating preventive or therapeutic interventions in patients with PM or PI, were identified through searching in electronic databases and in relevant journals. Reporting and methods were evaluated through an analysis of the risk of biases and quality score. Sub-analysis was performed in four subgroups: prevention of PM and PI, treatment of PM, and non-surgical and surgical treatment of PI. Thirty-two trials (29 RCT) were identified as accomplishing inclusion criteria after full-text reading. Seven focused on prevention, and among those dealing with therapy, six were related to PM and 19 related to PI therapy (10 non-surgical and 9 surgical). Analysis found that quality of reporting and methods was generally low and surrogate outcomes were often chosen. Current literature on PM and PI prevention and treatment does not allow extracting applicable clinical information. Quality of methods and reporting guidelines should be encouraged. In particular, ad hoc guidelines should be designed for peri-implant diseases.


Integration of virtual screening and susceptibility test to discover active-site subpocket-specific biogenic inhibitors of Helicobacter pylori shikimate dehydrogenase.
Shikimate dehydrogenase (HpSDH) (EC 22.214.171.124) is a key enzyme in the shikimate pathway of Helicobacter pylori (H. pylori), which catalyzes the NADPH-dependent reversible reduction of 3-dehydroshikimate to shikimate. Targeting HpSDH has been recognized as an attractive therapeutic strategy against H. pylori infection. Here, the catalytic active site in the crystal structure of HpSDH in complex with its substrate NADPH and product shikimate was examined in detail; the site can be divided into three spatially separated subpockets that separately correspond to the binding regions of shikimate, NADPH dihydronicotinamide moiety, and NADPH adenine moiety. Subsequently, a cascading protocol that integrated virtual screening and antibacterial test was performed against a biogenic compound library to identify biologically active, subpocket-specific inhibitors. Consequently, five, eight, and six promising compounds for, respectively, subpockets 1, 2, and 3 were selected from the top-100 docking-ranked hits, from which 11 compounds were determined to have high or moderate antibacterial potencies against two reference H. pylori strains, with MIC range between 8 and 93 μg/mL. It is found that the HpSDH active site prefers to accommodate amphipathic and polar inhibitors that consist of an aromatic core as well as a number of oxygen-rich polar/charged substituents such as hydroxyl, carbonyl, and carboxyl groups. Subpockets 1- and 2-specific inhibitors exhibit a generally higher activity than subpocket 3-specific inhibitors. Molecular dynamics simulations revealed an intense nonbonded network of hydrogen bonds, π-π stacking, and van der Waals contacts at the tightly packed complex interfaces of active-site subpockets with their cognate inhibitors, conferring strong stability and specificity to these complex systems. Binding energetic analysis demonstrated that the identified potent inhibitors can target their cognate subpockets with an effective selectivity over noncognate ones.


The effect of the messenger RNA concentration on the competitive inhibition of translation by cap-analogues.
Inhibition of translation of several mRNA species in a micrococcal nuclease treated reticulocyte lysate by cap analogues was compared with the competition between two mRNAs. Inhibition characteristics were very similar, only complete mRNA molecules inhibited at concentrations 150 times lower than m7 G5'ppp5'G. The inhibition of mRNA translation by cap analogues could be neutralized by the addition of extra mRNA in a manner predicted from the competitive nature of the inhibition by cap analogues.


DnaJ (Hsp40 protein) binding to folded substrate impacts KplE1 prophage excision efficiency.
Temperate phages mediate gene transfer and can modify the properties of their host organisms through the acquisition of novel genes, a process called lysogeny. The KplE1 prophage is one of the 10 prophage regions in Escherichia coli K12 MG1655. KplE1 is defective for lysis but fully competent for site-specific recombination. The TorI recombination directionality factor is strictly required for prophage excision from the host genome. We have previously shown that DnaJ promotes KplE1 excision by increasing the affinity of TorI for its site-specific recombination DNA target. Here, we provide evidence of a direct association between TorI and DnaJ using in vitro cross-linking assays and limited proteolysis experiments that show that this interaction allows both proteins to be transiently protected from trypsin digestion. Interestingly, NMR titration experiments showed that binding of DnaJ involves specific regions of the TorI structure. These regions, mainly composed of α-helices, are located on a surface opposite the DNA-binding site. Taken together, we propose that DnaJ, without the aid of DnaK/GrpE, is capable of increasing the efficiency of KplE1 excision by causing a conformational stabilization that allows TorI to adopt a more favorable conformation for binding to its specific DNA target.


Renal and liver functions and muscle injuries during training and after competition in Thai boxers.
To observe whether there are any injuries to muscle and deleterious effects on the liver and kidneys during training and after competition in Thai boxers. Serum levels of intracellular enzymes and specific markers in the urine were measured during training and after fighting in Thai boxers. During the training period, the activities of muscle enzymes were significantly increased whereas those of the liver enzymes and creatinine clearance were not changed. After a match, on the other hand, both liver and muscle enzyme activities were elevated but renal function was decreased. The training protocol for Thai boxers has virtually no deleterious effect on liver and renal function, but damage to skeletal muscle cells may occur. However, competition may cause muscle injury without any obvious damage to the liver and kidneys.


Frequency of β-Blocker Use Following Exacerbations of COPD in Patients with Compelling Indication for Use.
To assess the current use of β-blockers in patients with compelling indications for use, following the acute exacerbation of chronic obstructive pulmonary disease (COPD). We performed a multicenter retrospective observational study using data from all of the patients admitted to five institutions for an acute exacerbation of COPD. Patients were included if they were admitted for an acute exacerbation of COPD and had a compelling indication for the use of a β-blocker, defined as previous myocardial infarction or heart failure with left ventricular ejection fraction ≤40%. There were 396 patients meeting the criteria for inclusion in the study. The population was predominantly white men with myocardial infarction as the most prevalent compelling indication. On admission, 267 (67.4%) patients were receiving β-blockers, which increased to 278 (70.2%) at discharge. There were 118 (29.8%) patients discharged without β-blockers. Of the predictors tested, none were significantly predictive of a patient not receiving β-blockers upon discharge; however, home and in-hospital β-blockers reduced the likelihood of being discharged without a β-blocker. Of the 129 patients not receiving β-blockers prehospitalization, 23 (17.8%) were discharged with a new prescription for a β-blocker. Nearly one-third of patients with compelling indications for β-blockers were not prescribed the therapy at discharge.


Application of Raman spectroscopy to biological macromolecules.
The Raman spectra of biological macromolecules arise from molecular vibrations of either the backbone chains or the side chains. The frequencies of the Raman bands lie in a region between 200 cm-1 and 3000 cm-1. From certain frequencies of the vibrations of the backbone chains one can determine the conformation or secondary structure of a macromolecule. Thus for polypeptides and proteins the frequencies of the Amide I and Amide III vibrations allow one to determine the averge conformation of their backbone chain. In polynucleotides and nucleic acids, the frequency of the phosphate diester stretch of the phosphate furanose chain varies between 814 cm-1 for A conformation and 790 cm-1 for B conformation. Raman spectra of the bases in nucleic acids can be used to determine base stacking and hydrogen bonding interactions. Thus Raman spectroscopy is an important tool for determining the conformation structure of proteins and nucleic acids.


Safe zone for arthroscopic resection of the superomedial scapular border in the treatment of snapping scapula syndrome.
Arthroscopic resection of the superomedial corner of the scapula was performed in 20 fresh frozen cadaveric specimens to define clinically safe and useful margins for arthroscopic bony resection. The lateral border of resection was delineated by directing the burr either toward a specifically chosen arthroscopic resection target (ART) situated equidistant between the inferior scapular angle and scapular spine or more lateral to the inferior angle of the scapula. The minimum distances between the suprascapular notch and the lateral edge of the resection were 25 mm (average, 31 mm) in the ART group and 10 mm (average, 21 mm) in the inferior angle group (P < .01). From the results of our study, we recommend a safe zone for arthroscopic removal of bone and soft tissue from the superomedial corner of the scapula. The medial border is defined by the medial scapula and scapular spine; the lateral border is drawn between Bell's portal and the ART.


Priapism--a complication of home parenteral nutrition?
A case of priapism in a patient receiving home parenteral nutrition is reported. To our knowledge this is the first such case to occur in a patient receiving home parenteral nutrition. The literature is reviewed and the possible aetiology discussed.


Endogenous CRISPR-Cas mediated in situ genome editing: State-of-the-art and the road ahead for engineering prokaryotes.
The CRISPR-Cas systems have shown tremendous promise as heterologous tools for genome editing in various prokaryotes. However, the perturbation of DNA homeostasis and the inherent toxicity of Cas9/12a proteins could easily lead to cell death, which led to the development of endogenous CRISPR-Cas systems. Programming the widespread endogenous CRISPR-Cas systems for in situ genome editing represents a promising tool in prokaryotes, especially in genetically intractable species. Here, this review briefly summarizes the advances of endogenous CRISPR-Cas-mediated genome editing, covering aspects of establishing and optimizing the genetic tools. In particular, this review presents the application of different types of endogenous CRISPR-Cas tools for strain engineering, including genome editing and genetic regulation. Notably, this review also provides a detailed discussion of the transposon-associated CRISPR-Cas systems, and the programmable RNA-guided transposition using endogenous CRISPR-Cas systems to enable editing of microbial communities for understanding and control. Therefore, they will be a powerful tool for targeted genetic manipulation. Overall, this review will not only facilitate the development of standard genetic manipulation tools for non-model prokaryotes but will also enable more non-model prokaryotes to be genetically tractable.


Field estimates of the strength of interference between oystercatchers haematopus ostralegus.
The rate at which oystercatchers ate mussels was higher when few oystercatchers were present. The calculated strength of this interference (m) was immensely higher than is compatible with those models which assume interference is coused solely by the time predators waste when they encounter each other.


Purification, characterization, and kinetic analysis of a 55-kDa form of phosphatidylinositol 4-kinase from Saccharomyces cerevisiae.
A 55-kDa form of membrane-associated phosphatidylinositol 4-kinase (ATP:phosphatidylinositol 4-phosphotransferase, EC 184.108.40.206) was purified 10,166-fold from Saccharomyces cerevisiae. The purification procedure included solubilization of microsome membranes with 1% Triton X-100 followed by chromatography with DE52, hydroxylapatite I, Q-Sepharose, Mono Q, and hydroxylapatite II. The procedure resulted in a nearly homogeneous 55-kDa phosphatidylinositol 4-kinase preparation. The 55-kDa phosphatidylinositol 4-kinase and the previously purified 45-kDa phosphatidylinositol 4-kinase differed with respect to their amino acid composition, isoelectric points, and peptide maps. Furthermore, the two forms of phosphatidylinositol 4-kinase did not show an immunological relationship. Maximum 55-kDa phosphatidylinositol 4-kinase activity was dependent on magnesium (10 mM) or manganese (0.5 mM) ions and Triton X-100 at the pH optimum of 7.0. The activation energy for the reaction was 12 kcal/mol, and the enzyme was labile above 30 degrees C. The enzyme was inhibited by thioreactive agents, MgADP, and calcium ions. A detailed kinetic analysis of the purified enzyme was performed using Triton X-100/phosphatidylinositol-mixed micelles. 55-kDa phosphatidylinositol 4-kinase activity followed saturation kinetics with respect to the bulk and surface concentrations of phosphatidylinositol and followed surface dilution kinetics. The interfacial Michaelis constant (Km) and the dissociation constant (Ks) for phosphatidylinositol in the Triton X-100 micelle surface were 1.3 mol % and 0.035 mM, respectively. The Km for MgATP was 0.36 mM. 55-kDa phosphatidylinositol 4-kinase catalyzed a sequential reaction mechanism as indicated by the results of kinetic and isotopic exchange reactions. The enzyme bound to phosphatidylinositol before ATP and released phosphatidylinositol 4-phosphate before ADP. The enzymological and kinetic properties of the 55-kDa phosphatidylinositol 4-kinase differed significantly from those of the 45-kDa phosphatidylinositol 4-kinase. This may suggest that the two forms of phosphatidylinositol 4-kinase from S. cerevisiae are regulated differentially in vivo.


Ten-year clinical performance of zirconia posterior fixed partial dentures.
With the constantly increasing demand for metal-free solutions in dental therapy, numerous ceramic restorations have found their way into everyday clinical practice, but long-term clinical data are limited. The aim of this prospective clinical study was to evaluate three- and four-unit fixed partial dentures in the posterior region made of zirconium dioxide frameworks veneered with feldspathic porcelain after 10 years in clinical use. Based on the two studies published in 2009 and 2012, in which the all-ceramic FDPs were evaluated after 3 and 5 years of function, a clinical evaluation of a total of 17 restorations after 10 years with regard to their condition and long-term stability was carried out in the course of this study. The restorations were fabricated using feldspathic ceramic-veneered, yttria-stabilised, tetragonal zirconium dioxide as the framework material. The data collection was based on modified CDA criteria and included, for example, the shape, shade, surface condition and the success and survival rates of the restorations. The all-ceramic prostheses embodied excellent biocompatibility and colour reproduction. As a result of an increased incidence of chipping fractures, the success rate was 60%. The survival rate, however, was 88.2%, as 2 of the 17 restorations were lost. All-ceramic concepts for FDP constructions in the posterior region achieved satisfactory results in terms of durability after 10 years. The main problem was chipping, as has been generally recognised. Nevertheless, the materials were characterised by excellent aesthetics and biocompatibility, which ultimately makes them a good alternative to conventional restorative options. This study is registered in DRKS-German Clinical Trials Register with the register number DRKS00021743.


Neuropsychological deficits in childhood epilepsy syndromes.
Seizure disorders are relatively common in childhood, and the International League Against Epilepsy (ILAE) provides a hierarchical classification system to define seizure types. At the final level of classification, specific epilepsy syndromes are defined that represent a complex of signs and symptoms unique to an epilepsy condition. The present review discusses the issues related to several of these epilepsy syndromes in childhood, including those classified as generalized idiopathic epilepsies (e.g., childhood absence epilepsy, juvenile absence epilepsy, juvenile myoclonic epilepsy), focal epilepsies (benign rolandic epilepsy, occipital epilepsy, temporal lobe epilepsy, frontal lobe epilepsy) and the "epileptic encephalopathies," including Dravet's Syndrome, West Syndrome, Lennox-Gastaut Syndrome, Myoclonic Astatic Epilepsy, and Landau-Kleffner Syndrome. For each syndrome, the epidemiology, clinical manifestations, treatments, and neuropsychological findings are discussed.


The proteins of varicella-zoster-virus.
[35S] methionine-labelled Varicella-Zoster-Virus (VZV) was partly purified by sucrose density gradient centrifugation. Enveloped virus, checked by electron microscopy, was analyzed by high resolution polyacrylamide gel electrophoresis. A polypeptide pattern of at least 31 proteins could be identified on the fluorogram. The molecular weight of the proteins, estimated by co-electrophoresis with marker proteins, ranged from 18,000 to 240,000 daltons. Immunoprecipitation using a specific human antiserum showed at least 14 proteins.


Liquid crystal-based molecularly imprinted nanoparticles with low crosslinking for capillary electrochromatography.
In this work, molecularly imprinted polymer (MIP) nanoparticles were prepared at a low level of crosslinking in the presence of liquid crystalline monomer as physical crosslinker to replace the most of chemical crosslinker. The imprinted nanoparticles against d-zopiclone were synthesized by precipitation polymerization using a mixture of methacrylic acid, ethylene glycol dimethacrylate, and liquid crystalline monomers. The resulting d-zopiclone imprinted nanoparticles were evaluated in capillary electrochromatography by partial filling technique. The resolution of enantiomer separation achieved on the d-zopiclone-imprinted nanoparticles was up to 3.29 and the column efficiency of zopiclone enantiomer (up to 66,900 plates/m) with good peak symmetry was obtained. The imprinted nanoparticles prepared in the presence of liquid crystalline monomer can retain affinity and specificity for template even when prepared with a level of cross-linker as low as 5%.


Inhibition of p38 and ERK MAP kinases blocks endotoxin-induced nitric oxide production and differentially modulates cytokine expression.
Mitogen-activated protein kinases (MAPKs) are thought to have a critical role in lipopolysaccharide (LPS)-induced immune responses but the molecular mechanisms underlying the mediation of these signaling are not clear. The roles of p38 and extracellular signal-regulated kinase (ERK) in the regulation of nitric oxide (NO) and proinflammatory cytokine expression in J774A.1 macrophages in response to LPS were examined. Specific inhibitors for p38 and ERK, SB203580 and PD98059, respectively, were used. LPS (30ng/ml) activated inducible nitric oxide synthase (iNOS), subsequent NO production, and gene expression for tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6, and IL-12. Treatment of cultures with SB203580 increased LPS-induced reactive oxygen species (ROS) production, whereas both SB203580 and PD98059 decreased LPS-induced NO production. Concomitant decreases in the expression of iNOS mRNA and protein were detected. SB203580 and PD98059 decreased LPS-induced gene expression of IL-1beta and IL-6. SB203580 increased LPS-induced expression of TNF-alpha and IL-12; PD98059 had no effect on these cytokines. Results indicated that both p38 and ERK pathways are involved in LPS-stimulated NO synthesis and the expression of IL-1beta and IL-6. p38 signaling pathway is involved in LPS-induced ROS, TNF-alpha and IL-12 production.


Photoactivation Reduces Side-Chain Dynamics of a LOV Photoreceptor.
We used neutron-scattering experiments to probe the conformational dynamics of the light, oxygen, voltage (LOV) photoreceptor PpSB1-LOV from Pseudomonas putida in both the dark and light states. Global protein diffusion and internal macromolecular dynamics were measured using incoherent neutron time-of-flight and backscattering spectroscopy on the picosecond to nanosecond timescales. Global protein diffusion of PpSB1-LOV is not influenced by photoactivation. Observation-time-dependent global diffusion coefficients were found, which converge on the nanosecond timescale toward diffusion coefficients determined by dynamic light scattering. Mean-square displacements of localized internal motions and effective force constants, <k'>, describing the resilience of the proteins were determined on the respective timescales. Photoactivation significantly modifies the flexibility and the resilience of PpSB1-LOV. On the fast, picosecond timescale, small changes in the mean-square displacement and <k'> are observed, which are enhanced on the slower, nanosecond timescale. Photoactivation results in a slightly larger resilience of the photoreceptor on the fast, picosecond timescale, whereas in the nanosecond range, a significantly less resilient structure of the light-state protein is observed. For a residue-resolved interpretation of the experimental neutron-scattering data, we analyzed molecular dynamics simulations of the PpSB1-LOV X-ray structure. Based on these data, it is tempting to speculate that light-induced changes in the protein result in altered side-chain mobility mostly for residues on the protruding Jα helix and on the LOV-LOV dimer interface. Our results provide strong experimental evidence that side-chain dynamics play a crucial role in photoactivation and signaling of PpSB1-LOV via modulation of conformational entropy.


[3H]5-HT binding in post-mortem human cerebral cortex: methodological considerations.
The effects of different membrane preparations and assay conditions on [3H]5-HT binding to post-mortem human cortical tissue was studied. Optimal binding necessitated thorough removal of endogenous 5-HT and this was achieved either by hypotonic lysis or by preincubation of the membranes at 37 degrees C. Calcium chloride (4 mM) increased specific [3H]5-HT binding. The further addition of ascorbic acid (5.7 mM) or ascorbic acid and clorgyline (10 microM) reduced specific [3H]5-HT binding.


Stimulation of cell invasion by the Golgi Ion Channel GAAP/TMBIM4 via an H2O2-Dependent Mechanism.
The mechanisms by which the Golgi apparatus (GA) impacts on cell invasion are poorly understood. The human Golgi Anti-Apoptotic Protein (hGAAP, also known as TMBIM4) is a highly conserved Golgi cation channel that modulates intracellular Ca2+ fluxes. Human GAAP is expressed in all human tissues, is essential for cell viability and provides resistance against a range of apoptotic stresses. Furthermore, hGAAP enhances adhesion and cell migration by increasing the turnover of focal adhesions due to activation of store-operated Ca2+ entry. Here, we describe a GA-derived mechanism that controls cell invasion. The overexpression of hGAAP stimulates 3-dimensional proteolytic cell invasion by a mechanism that is dependent on the accumulation of intracellular hydrogen peroxide, which might be produced by the hGAAP-dependent stimulation of mitochondrial respiration. These findings provide new insight into the complex mechanisms by which Ca2+ and reactive oxygen species signaling contribute to cell invasion and to the role of the GA in these processes.


Chemoradiotherapy with temozolomide after high-dose methotrexate for primary CNS lymphoma: a multicenter phase I study of a response-adapted strategy.
This study aimed to define the maximum tolerated dose (MTD) of temozolomide (TMZ) concurrent with radiotherapy (RT) after high-dose methotrexate (HD-MTX) for newly diagnosed primary central nervous system lymphoma (PCNSL). Adult patients with PCNSL were treated according to a response-adapted strategy. HD-MTX (3.5 g/m2) was followed by concomitant RT and escalating TMZ (50-60-75 mg/m2/day, 5 days/week). The total radiation dose was modulated according to the patient's response to HD-MTX. All patients received 30 Gy to the whole brain plus leptomeninges to C2, including the third posterior of the orbital cavity (clinical target volume 2; CTV2), plus 6, 10, or 16 Gy to the primary site, including the residual mass (CTV1), if a complete response (CR), partial response (PR)/stable disease (SD), or progressive disease (PD) was observed, respectively. Acute toxicities were graded according to the RTOG-EORTC criteria. Dose-limiting toxicity (DLT) was defined as grade 4 hematological toxicity or grade 3-4 hepatic toxicity, although 75 mg/m2/day was the maximum dose regardless of DLT. Neurocognitive function was evaluated using the Mini-Mental State Examination. Three patients were enrolled at each TMZ dose level (total = 9 patients). Twelve lesions were treated. Six patients received 2 cycles of HD-MTX, while 3 received only 1 cycle because of hepatic or renal toxicity. All patients completed chemoradiotherapy without interruptions. No DLT events were recorded. TMZ appears to be tolerable at a dose of 75 mg/m2/day when administered concomitantly with radiotherapy and after HD-MTX.