The exocyst complex and intracellular vesicles mediate soluble protein trafficking to the primary cilium.

The efficient transport of proteins into the primary cilium is a crucial step for many signaling pathways. Dysfunction of this process can lead to the disruption of signaling cascades or cilium assembly, resulting in developmental disorders and cancer. Previous studies on the protein delivery to the cilium were mostly focused on the membrane-embedded receptors. In contrast, how soluble proteins are delivered into the cilium is poorly understood. In our work, we identify the exocyst complex as a key player in the ciliary trafficking of soluble Gli transcription factors. In line with the known function of the exocyst in intracellular vesicle transport, we demonstrate that soluble proteins, including Gli2/3 and Lkb1, can use the endosome recycling machinery for their delivery to the primary cilium. Finally, we identify GTPases: Rab14, Rab18, Rab23, and Arf4 that are involved in vesicle-mediated Gli protein ciliary trafficking. Our data pave the way for a better understanding of ciliary transport and uncover transport mechanisms inside the cell.

MRCKα/ß positively regulates Gli protein activity.

Posttranslational modifications (PTMs) are key regulatory events for the majority of signaling pathways. Transcription factors are often phosphorylated on multiple residues, which regulates their trafficking, stability, or transcriptional activity. Gli proteins, transcription factors that respond to the Hedgehog pathway, are regulated by phosphorylation, but the sites and the kinases involved have been only partially described. We identified three novel kinases: MRCKα, MRCKß, and MAP4K5 which physically interact with Gli proteins and directly phosphorylate Gli2 on multiple sites. We established that MRCKα/ß kinases regulate Gli proteins, which impacts the transcriptional output of the Hedgehog pathway. We showed that double knockout of MRCKα/ß affects Gli2 ciliary and nuclear localization and reduces Gli2 binding to the Gli1 promoter. Our research fills a critical gap in our understanding of the regulation of Gli proteins by describing their activation mechanisms through phosphorylation.

Analyzing Complex Problem Solving by Dynamic Brain Networks.

Complex problem solving is a high level cognitive task of the human brain, which has been studied over the last decade. Tower of London (TOL) is a game that has been widely used to study complex problem solving. In this paper, we aim to explore the underlying cognitive network structure among anatomical regions of complex problem solving and its subtasks, namely planning and execution. A new computational model for estimating a brain network at each time instant of fMRI recordings is proposed. The suggested method models the brain network as an Artificial Neural Network, where the weights correspond to the relationships among the brain anatomic regions. The first step of the model is preprocessing that manages to decrease the spatial redundancy while increasing the temporal resolution of the fMRI recordings. Then, dynamic brain networks are estimated using the preprocessed fMRI signal to train the Artificial Neural Network. The properties of the estimated brain networks are studied in order to identify regions of interest, such as hubs and subgroups of densely connected brain regions. The representation power of the suggested brain network is shown by decoding the planning and execution subtasks of complex problem solving. Our findings are consistent with the previous results of experimental psychology. Furthermore, it is observed that there are more hubs during the planning phase compared to the execution phase, and the clusters are more strongly connected during planning compared to execution.

Subgap dynamics of double quantum dot coupled between superconducting and normal leads.

Dynamical processes induced by the external time-dependent fields can provide valuable insight into the characteristic energy scales of a given physical system. We investigate them here in a nanoscopic heterostructure, consisting of the double quantum dot coupled in series to the superconducting and the metallic reservoirs, analyzing its response to (i) abrupt bias voltage applied across the junction, (ii) sudden change of the energy levels, and imposed by (iii) their periodic driving. We explore subgap properties of this setup which are strictly related to the in-gap quasiparticles and discuss their signatures manifested in the time-dependent charge currents. The characteristic multi-mode oscillations, their beating patters and photon-assisted harmonics reveal a rich spectrum of dynamical features that might be important for designing the superconducting qubits.

Morphological description of the alimentary tract of Geoicautricularia (Passerini, 1856) (Insecta, Hemiptera, Eriosomatinae).

Existing literature data report the lack of stomach and crenated intestine in the aphid species Geoicasetulosa (Passerini, 1860), a representative of subfamily Eriosomatinae. This odd anatomical feature seemed remarkable, due to the presence of fully developed intestine in closely related genera and mutualistic relationship with ants of this genus. The study aimed at repeated anatomical research of Geoicautricularia (Passerini 1856), in order to confirm what seemed to be a generic feature. Standard histological methods were applied, with addition of oblique light microscopy, fluorescence microscopy and confocal laser scanning microscopy. The results indicated the existence of a fully developed intestine, with broad sac-shaped stomach and loops of the crenated intestine. The general anatomy of the alimentary tract of G. utricularia resembles that of other representatives of the tribe Fordini. Also well-developed rectal gland is present, most probably playing a role in modifying the carbohydrate composition of excreted honeydew.

Can Helicobacter pylori be eradicated with high-dose proton pump inhibitor in extensive metabolizers with the CYP2C19 genotypic polymorphism?

Proton pump inhibitors (PPI) metabolism and pharmacokinetics are regulated by cytochrome P450 enzymes in the liver. Cytochrome P450 2C19 (CYP2C19) polymorphism plays an import role in the metabolism of PPIs. The three possible genotypes for CYP2C19 each has a distinct effect on the pharmacodynamics of PPIs. Homozygote extensive metabolizers (HomEM) are the most frequent genotype and have two wild-types (non-mutant) (*1/*1) alleles. HomEM is associated with increased enzyme activity, which increases the rate of PPI metabolism. Intragastric pH, which is required for eradication, is lowest in HomEM. In HomEMs, an insufficient increase in intragastric pH results in decreased anti-Helicobacter pylori (HP) efficacy of the antibiotics and, therefore, lower eradication rates. We determined whether the HP eradication rate would increase after high-dose PPI treatment of extensive PPI metabolizers who had been treated unsuccessfully with a standard PPI dose. In our report, increasing the PPI dosage in patients with genotype polymorphisms may be effective on eradication rates. Eradication rates are directly affected by CYP2C19 polymorphisms, and eradication treatments should be planned considering such genotypic polymorphisms. Hence, CYP2C19 genotyping prior to treatment may facilitate determination of the optimum PPI dose to improve the therapeutic outcome. However, further researches are required to confirm this hypothesis.

Steroid-refractory inflammatory bowel disease is a risk factor for CMV infection.

OBJECTIVE: Patients with inflammatory bowel disease (IBD) show increased the prevalence of cytomegalovirus (CMV) infection due to the severity of the disease and the immunosuppressive treatments they receive. The aim of this study was to determine the prevalence of CMV infection in IBD patients and identify the risk factors for CMV infection with different demographic characteristics in IBD patients. PATIENTS AND METHODS: We enrolled 85 patients diagnosed with IBD (43 with ulcerative colitis (UC) and 42 with Crohn's disease (CD)) in this prospective study. The clinical disease activities of UC and CD were assessed using Truelove-Witts and Crohn's disease activity index (CDAI). CMV infection was assessed by detection of DNA using real-time polymerase chain reaction (PCR) in blood samples and quantitative PCR in colonic biopsy specimens and by detection of inclusion bodies using hematoxylin-eosin staining. RESULTS: Thirteen patients with IBD exhibited concomitant CMV infection. CMV infection was not detected in any of the patients in remission. Viral loads measured in the colonic mucosa of infected patients ranged from 800-7000 genome copies/mL total extracted DNA. The mean serum CMV DNA level was 1694 ± 910 copies/mL (range: 800-3800). The rate of steroid resistance in CMV-positive cases was significantly higher than that in CMV-negative cases (p = 0.001). CD with acute exacerbation was a risk factor for CMV disease (p = 0.04). All of the CMV-positive patients received immunosuppressive treatments. CONCLUSIONS: CMV infection should be suspected in steroid-resistant UC and CD. Antiviral treatment improved the clinical outcome in steroid-resistant IBD cases with serum CMV DNA levels above 1000 copies/mL.

Effect of cytochrome P450 2C19 polymorphisms on the Helicobacter pylori eradication rate following two-week triple therapy with pantoprazole or rabeprazole.

OBJECTIVE: Cytochrome P450 2C19 (CYP2C19) polymorphisms play an important role in the metabolism of proton pump inhibitors. Rabeprazole is primarily metabolized via non-enzymatic pathways. In this study, we determined whether rabeprazole- and pantoprazole-based eradication treatments were influenced by CYP2C19 polymorphisms. PATIENTS AND METHODS: A total of 200 patients infected with Helicobacter pylori were treated with either 40 mg of pantoprazole or 20 mg of rabeprazole plus 500 mg of clarithromycin, 1000 mg of amoxicillin twice daily for 2 weeks. CYP2C19 genotype status was determined by Polymerase Chain Reaction (PCR)-restriction-fragment-length polymorphism. The genotypes of cytochrome P450 2C19 were classified as homozigote extensive metabolizer (HomEM), heterozigote metabolizer (HetEM) and poor metabolizer (PM). The CYP2C19 genotype of all patients, the effectiveness of the treatment, the effect of the genotypic polymorphism on the treatment were assessed. RESULTS: The frequencies of HotEM, HetEM, PM were 78%, 19.5% and 2.5%, respectively. 48% (n = 96) of the patients received treatment with rabeprazole and 52% (n = 104) with pantoprazole. The eradication rate was 64.7% for HomEM, 79.4% for HetEM, 100% for PM (p = 0.06). In HetEM, PM, are considered as a single group, the eradication rates were higher in patients with the HetEM and PM (HetEM+PM) genotypes than in those with the wild-type genotype (81.8 vs. 64.7% p = 0.031). Among the patients treated with rabeprazole, the eradication rates were significantly lower in those with the HomEM genotype than in those with the HetEM+PM genotypes (60% vs. 85.7% p = 0.023). CONCLUSIONS: The genotypic polymorphism is effective on the rate of eradication. Eradication treatment rate with rabeprazole is influenced by CYP2C19 genotype.

Infliximab in inflammatory bowel disease: attention to adverse events.

OBJECTIVE: To assess the efficacy and adverse effects of infliximab in patients with Crohn's disease and ulcerative colitis who are resistant to conventional therapy or having fistulising type Crohn's disease. PATIENTS AND METHODS: The patients with a diagnosis of inflammatory bowel disease received infliximab between 2007 and 2009 were followed-up prospectively. Infliximab 5 mg/kg was given at week 0, 2, 6, and every 8 weeks thereafter. Early and late adverse events occurring during the treatment were recorded for each patient. RESULTS: There were 36 patients [mean age 35±12, 17 male] included in the study. Thirty-two (88%) patients were receiving concomitant long-term immunosuppressive therapy. Complete or partial response was obtained in 75% of all patients. At least one adverse event was observed in 10 (28%) patients. Anaphylaxis was seen in 2 (6%) patients, mild acute infusion reaction in 2 (6%) patients, hypotension in 2 (6%) patients, respiratory distress in 2 (6%) patients, skin rash and eruptions in 2 (6%) patients, one hypertension (3%) and one (3%) tightness in the chest. Treatment was continued in all except patients with anaphylaxis. No infection, tumour or cases of death were observed. CONCLUSIONS: Several adverse events might be observed in patients who receive infliximab. Care should be given to patients whom treatment was restarted after a break in regard to anaphylaxis. No serious adverse event was observed during infliximab treatment except allergic events.

Treatment failure may lead to accelerated fibrosis progression in patients with chronic hepatitis C.

Chronic hepatitis C (CHC) patients with treatment failure (TF) remain at risk of continuing fibrosis progression. However, it has not been investigated whether there is an increased risk of accelerated fibrosis progression after failed interferon-based therapy. We aimed to investigate long-term influence of TF on fibrosis progression compared with untreated patients with CHC. We studied 125 patients with CHC who underwent paired liver biopsies from 1994 to 2012. Patients with advanced fibrosis were excluded from the analysis. Sixty-three patients had TF, and 62 patients were treatment-naïve (TN). Annual fibrosis progression rate (FPR) was calculated, and significant fibrosis progression (SFP) was defined as ≥ 2 stage increase in fibrosis during follow-up. Multiple regression analyses were performed to find out independent predictors of FPR and SFP. Demographic characteristics and duration between paired liver biopsies were similar in TF and TN groups. Baseline alanine aminotransferase and gamma-glutamyl transferase (GGT) levels (71 ± 31 vs 47 ± 22, P < 0.001 and 49 ± 39 vs 36 ± 28, P = 0.027, respectively), baseline mean fibrosis stage (2.2 ± 0.7 vs 1.9 ± 0.7, P = 0.018) and histologic activity index (6.3 ± 1.9 vs 4.3 ± 1.6, P < 0.001) were higher in the TF group compared with the TN group. In regression analyses, the strongest independent predictor of fibrosis progression was the GGT level (OR: 1.03, 95%CI 1.01-1.5, P < 0.001). Treatment experience (OR: 5.97, 95%CI 1.81-19.7, P = 0.003) also appeared as an independent predictor of both FPR and SFP. Failed interferon-based CHC treatment may lead to accelerated FPR in the long-term compared with the natural course.

Management of factor VII-deficient patients undergoing joint surgeries--preliminary results of locally developed treatment regimen.

Inherited factor VII (FVII) deficiency is a rare coagulation disorder with variable haemorrhagic manifestations. In severely affected cases spontaneous haemarthroses leading to advanced arthropathy have been observed. Such cases may require surgery. Therapeutic options for bleeding prevention in FVII deficient patients undergoing surgery comprise various FVII preparations but the use of recombinant activated factor VII (rFVIIa) seems to be the treatment of choice. To present the outcome of orthopaedic surgery under haemostatic coverage of rFVIIa administered according to the locally established treatment regimen in five adult patients with FVII baseline plasma levels below 10 IU dL(-1). Two patients required total hip replacement (THR); three had various arthroscopic procedures. Recombinant activated factor VII was administered every 8 h on day of surgery (D0) followed by every 12-24 h for the subsequent 9-14 days, depending on the type of surgery. Factor VII plasma coagulation activity (FVII:C) was determined daily with no predefined therapeutic target levels. Doses of rFVIIa on D0 ranged from 18 to 37 µg kg(-1) b.w. and on the subsequent days--from 13 to 30 µg kg(-1) b.w. Total rFVIIa dose per procedure ranged from 16 to 37.5 mg, and the total number of doses per procedure was 16-31. None of our patients developed excessive bleeding including those in whom FVII:C trough levels returned nearly to the baseline level on the first post-op day. Preliminary results demonstrate that rFVIIa administered according to our treatment regimen is an effective and safe haemostatic agent for hypoproconvertinaemia patients undergoing orthopaedic surgery.

Apolipoprotein E-e4, processing speed, and white matter volume in a genetically enriched sample of midlife adults.

Healthy midlife children of a parent with Alzheimer's disease ([AD] N = 23; 9 male) participated in neuropsychological testing, and magnetic resonance imaging (MRI) of brain volumetrics were obtained. In all, 35% of the sample were apolipoprotein E (ApoE)-e4 positive (n = 8; 5 male). The ApoE-e4 group exhibited significantly slower performances on an executive function and processing speed measure and had less white matter volume than the non-ApoE-e4 group. Lesser white matter volume was significantly correlated with slower processing speed. Processing speed and changes in white matter volume might be indicators of preclinical decline in AD.

Phenotype-genotype correlation in eight Polish patients with inherited Factor XIII deficiency: identification of three novel mutations.

Inherited factor XIII (FXIII) deficiency is known as one of the most rare blood coagulation disorder in humans. In the present study, phenotype and genotype of eight FXIII deficient Polish patients from five unrelated families were compared. The patients presented with a severe phenotype demonstrated by a high incidence of intracerebral haemorrhages (seven of eight patients), haemarthrosis (six patients) and bleeding due to trauma (five patients). Introduction of regular substitution with FXIII concentrate prevented spontaneous bleeding in seven patients. In all patients, mutations within the F13A gene have been identified revealing four missense mutations (Arg77Cys, Arg260Cys, Ala378Pro, Gly420Ser), one nonsense mutation (Arg661X), one splice site mutation (IVS5-1 G>A) and one small deletion (c.499-512del). One homozygous large deletion involving exon 15 was detected by failure of PCR product. The corresponding mutations resulted in severely reduced FXIII activity and FXIII A-subunit antigen concentration, while FXIII B-subunit antigen remained normal or mildly decreased. Structural analysis demonstrated that the novel Ala378Pro mutation may cause a disruption of the FXIII catalytic triad leading to a non-functional protein which presumably undergoes premature degradation. In conclusion, the severe phenotype with high incidence of intracranial bleeding and haemarthrosis was in accordance with laboratory findings on FXIII and with severe molecular defects of the F13A gene.

[Predictive values of peak expiratory flow measured with mechanical meters for Polish children and adolescents between 7-19 years of age]. / Wartosci nalezne szczytowego przeplywu wydechowego mierzonego miernikami mechanicznymi dla polskiej populacji dzieci i mlodziezy w wieku 7-19 LAT.

The aim of this study was to evaluate the predicted values of pack expiratory flow for the Polish children and youth by means of measures Mini Wright and Personal Best. The accounts were based on a group of 86 boys and 110 girls. It was established that it was necessary to state different predicted values for different measures and that an optimum model of regression for counting of predicted values of PEFR in relation to body height for the Polish children and youth was an exponential model. Model indices for both measures were set up separately for boys and girls.

[The transfer of hepatitis B virus (HBV) from HBsAg(+) mothers to their babies]. / HBsAg (+) annelerden dogan bebeklere hepatit B virusunun (HBV) geçis sikligi.

In this study, Hepatitis B surface antigen (HBsAg) was detected in 3.1% (19/602) of mothers just after delivery. HBsAg positivity was detected in 31% (6/19) in cord blood of newborn infants born from HBsAg positive mothers. Six months later, 15 of the HBsAg positive mothers and their children were screened serologically. HBsAg positivity persisted in only one child (1/15, 6.6%).