CDKL5 expression is modulated during neuronal development and its subcellular distribution is tightly regulated by the C-terminal tail

Mutations in the human X-linked cyclin-dependent kinase-like 5 (CDKL5) gene have been identified in patients with Rett syndrome (RTT), West syndrome, and X-linked infantile spasms, sharing the common feature of mental retardation and early seizures. CDKL5 is a rather uncharacterized kinase, but its involvement in RTT seems to be explained by the fact that it works upstream of MeCP2, the main cause of Rett syndrome. To understand the role of this kinase for nervous system functions and to address if molecular mechanisms are involved in regulating its distribution and activity, we studied the ontogeny of CDKL5 expression in developing mouse brains by immunostaining and Western blotting. The expression profile of CDKL5 was compared with that of MeCP2. The two proteins share a general expression profile in the adult mouse brain, but CDKL5 levels appear to be highly modulated at the regional level. Its expression is strongly induced in early postnatal stages, and in the adult brain CDKL5 is present in mature neurons, but not in astroglia. Interestingly, the presence of CDKL5 in the cell nucleus varies at the regional level of the adult brain and is developmentally regulated. CDKL5 shuttles between the cytoplasm and the nucleus and the C-terminal tail is involved in localizing the protein to the cytoplasm in a mechanism depending on active nuclear export. Accordingly, Rett derivatives containing disease-causing truncations of the C terminus are constitutively nuclear, suggesting that they might act as gain of function mutations in this cellular compartment

High-resolution tracking in a GEM-Emulsion detector

SHiP (Search for Hidden Particles) is a beam dump experiment proposed at the CERN SPS aiming at the observation of long lived particles very weakly coupled with ordinary matter mostly produced in the decay of charmed hadrons. The beam dump facility of SHiP is also a copious factory of neutrinos of all three kinds and therefore a dedicated neutrino detector is foreseen in the SHiP apparatus. The neutrino detector exploits the Emulsion Cloud Chamber technique with a modular structure, alternating walls of target units and planes of electronic detectors providing the time stamp to the event. GEM detectors are one of the possible choices for this task. This paper reports the results of the first exposure to a muon beam at CERN of a new hybrid chamber, obtained by coupling a GEM chamber and an emulsion detector. Thanks to the micrometric accuracy of the emulsion detector, the position resolution of the GEM chamber as a function of the particle inclination was evaluated in two configurations, with and without the magnetic fiel

Functional consequences of mutations in CDKL5, an X-linked gene involved in infantile spasms and mental retardation

Mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene have been identified in patients with Rett syndrome, West syndrome, and X-linked infantile spasms sharing the common features of generally intractable early seizures and mental retardation. Disease-causing mutations are distributed in both the catalytic domain and in the large COOH terminus. In this report, we examine the functional consequences of some Rett mutations of CDKL5 together with some synthetically designed derivatives useful to underline the functional domains of the protein. The mutated CDKL5 derivatives have been subjected to in vitro kinase assays and analyzed for phosphorylation of the TEY (Thr-Glu-Tyr) motif within the activation loop, their subcellular localization, and the capacity of CDKL5 to interact with itself. Whereas wild-type CDKL5 autophosphorylates and mediates the phosphorylation of the methyl-CpG-binding protein 2 (MeCP2) in vitro, Rett-mutated proteins show both impaired and increased catalytic activity suggesting that a tight regulation of CDKL5 is required for correct brain functions. Furthermore, we show that CDKL5 can self-associate and mediate the phosphorylation of its own TEY (Thr-Glu-Tyr) motif. Eventually, we show that the COOH terminus regulates CDKL5 properties; in particular, it negatively influences the catalytic activity and is required for its proper sub-nuclear localization. We propose a model in which CDKL5 phosphorylation is required for its entrance into the nucleus whereas a portion of the COOH-terminal domain is responsible for a stable residency in this cellular compartment probably through protein-protein interactions

Ig Glycosylation in Ulcerative Colitis: It’s Time for New Biomarkers

Background: Ulcerative colitis (UC) is a chronic relapsing disease, which needs a continue monitoring, especially during biological therapies. An increasing number of patients is treated with anti-Tumor Necrosis factor (TNF) drugs, and current research is focalized to identify biomarkers able to monitor the disease and to predict therapeutic outcome. Methods: We enrolled consecutive UC patients treated with anti-TNF, naïve to biologic drugs. Therapeutic outcome was evaluated after 54 weeks of treatment in terms of clinical remission (Partial Mayo Score -PMS- <2) and mucosal healing (Mayo Endoscopic Score <2). On serum samples collected at baseline and after 54 weeks of treatment, a Lectin-based ELISA assay was performed, and specific glycosylation patterns were evaluated by biotin-labelled lectins. We have also collected 21 healthy controls (NHS) samples, age and sex-matched. Results: Out of 44 UC patients enrolled, 22 achieved clinical remission and mucosal healing after 54 weeks. At baseline, when Protein A was used as coating, UC patients non-responders showed a reduced reactivity to Jacalin (JAC) in comparison with NHS (p = 0.04). After one year of treatment, a decrease in JAC binding was seen only in responders, in comparison with baseline (p = 0.04). When JAC binding was tested selecting IgG by means of Fab anti-IgG Fab, UC patients displayed an increased reactivity after anti-TNF therapy (p < 0,0001 vs controls). At baseline, PMS inversely correlates with JAC binding when Fab anti-IgG Fab was used in solid phase (r2 = 0,2211; p = 0,0033). Patients with higher PMS at baseline (PMS ≥5) presented lower binding capacity for JAC in comparison with NHS and with lower PMS patients (p = 0,0135 and p = 0,0089, respectively). Conclusion: Ig glycosylation was correlated with clinical and endoscopic activity in patients with UC. JAC protein A-selected Ig showed a possible role in predicting therapeutic effectiveness. If these data would be confirmed, Ig glycosylation could be used as biomarker in UC

Oral sucrosomial iron is as effective as intravenous ferric carboxy‐maltose in treating anemia in patients with ulcerative colitis

Anemia is a frequent complication of ulcerative colitis, and is frequently caused by iron deficiency. Oral iron supplementation displays high rates of gastrointestinal adverse effects. However, the formulation of sucrosomial iron (SI) has shown higher tolerability. We performed a prospective study to compare the effectiveness and tolerability of oral SI and intravenous ferric carboxy‐maltose (FCM) in patients with ulcerative colitis in remission and mild‐to‐moderate anemia. Patients were randomized 1:1 to receive 60 mg/day for 8 weeks and then 30 mg/day for 4 weeks of oral SI or intravenous 1000 mg of FCM at baseline. Hemoglobin and serum levels of iron and ferritin were assessed after 4, 8, and 12 weeks from baseline. Hemoglobin and serum iron increased in both groups after 4 weeks of therapy, and remained stable during follow up, without significant treatment or treatment‐by‐time interactions (p = 0.25 and p = 0.46 for hemoglobin, respectively; p = 0.25 and p = 0.26 for iron, respectively). Serum ferritin did not increase over time during SI supplementation, while it increased in patients treated with FCM (treatment effect, p = 0.0004; treatment‐bytime interaction effect, p = 0.0002). Overall, this study showed that SI and FCM displayed similar effectiveness and tolerability for treatment of mild‐to‐moderate anemia in patients with ulcerative colitis under remission

Addition of Thirteen Genes to the Porcine Comparative Gene Map Reveals New Regions of Conserved Synteny

Thirteen genes were mapped to the porcine genome by using either linkage mapping of the PiGMaP families (eight genes) or typing of a porcine somatic cell hybrid panel (12 genes). The genes were chosen from interesting locations in the human genome. The physical gene assignments to pig chromosomes (SSC) with corresponding human chromosome (HSA) locations include the following: FGF7 (HSA15), MADH4 (HSA18), and MC4R (HSA18) to SSC1, RXRB (HSA6), and SSTR1 (HSA14) to SSC7, UCP1 (HSA4) to SSC8, PGR (HSA11) to SSC9, TTN (HSA2) and ANT1 (HSA4) to SSC15, GRIA1 (HSA5) to SSC16, AR (HSA-X), and GRIA3 (HSA-X) to SSC-X. Additionally, CD59 (HSA11) was linkage mapped to SSC2. The majority of the assignments confirm results from bidirectional chromosome painting (4). A rearrangement in gene order was detected within the region of correspondence between SSC1 and HSA15. Two assignments were made that were not expected from the painting results (MC4R and GRIA1) and one assignment of a gene from a region where the painting study was not informative (ANT1)

A low-FODMAP diet for irritable bowel syndrome: Some answers to the doubts from a long-term follow-up

A low-FODMAP (fermentable oligosaccharides, disaccharides, monosaccharides and polyols) diet (LFD) is a possible therapy for irritable bowel syndrome (IBS). This study investigates the short-and long-term efficacy and nutritional adequacy of an LFD and the patients’ long-term acceptability. Patients’ adherence and ability to perceive the “trigger” foods were also evaluated. Seventy-three IBS patients were given an LFD (T0) and after 2 months (T1), 68 started the reintroduction phase. At the end of this period (T2), 59 were advised to go on an Adapted Low-FODMAP Diet (AdLFD) and 41 were evaluated again after a 6–24 month follow-up (T3). At each time, questionnaires and Biolectrical Impedance Vector Analysis (BIVA) were performed. The LFD was effective in controlling digestive symptoms both in the short-and long-term, and in improving quality of life, anxiety and depression, even if some problems regarding acceptability were reported and adherence decreased in the long term. The LFD improved the food-related quality of life without affecting nutritional adequacy. When data collected at T0 were compared with those collected at T2, the perception of trigger foods was quite different. Even if some problems of acceptability and adherence are reported, an LFD is nutritionally adequate and effective in improving IBS symptoms also in the long term

First determination of the one-proton induced Non-Mesonic Weak Decay width of p-shell {\Lambda}-Hypernuclei

Previous studies of proton and neutron spectra from Non-Mesonic Weak Decay of eight Lambda-Hypernuclei (A = 5-16) have been revisited. New values of the ratio of the two-nucleon and the one-proton induced decay widths, Gamma_2N/Gamma_p, are obtained from single proton spectra, Gamma_2N/Gamma_p = 0.50 +/- 0.24, and from neutron and proton coincidence spectra, Gamma_2N/Gamma_p = 0.36 +/- 0.14stat +0.05sys -0.04sys , in full agreement with previously published ones. With these values, a method is developed to extract the one-proton induced decay width in units of the free Lambda decay width, Gamma_p/Gamma_Lambda, without resorting to Intra Nuclear Cascade models but by exploiting only experimental data, under the assumption of a linear dependence on A of the Final State Interaction contribution. This is the first systematic determination ever done and it agrees within the errors with recent theoretical calculations.Comment: 16 pages, 3 figures, 2 table