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

A simple and reliable methodology to detect egg white in art samples

A protocol for a simple and reliable dot-blot immunoassay was developed and optimized to test work of art samples for the presence of specific proteinaceus material (i.e. ovalbumin-based). The analytical protocol has been extensively set up with respect, among the other, to protein extraction conditions, to densitometric analysis and to the colorimetric reaction conditions. Feasibility evaluation demonstrated that a commercial scanner and a free image analysis software can be used for the data acquisition and elaboration, thus facilitating the application of the proposed protocol to commonly equipped laboratories and to laboratories of museums and conservation centres. The introduction of method of standard additions in the analysis of fresh and artificially aged laboratory-prepared samples, containing egg white and various pigments, allowed us to evaluate the matrix effect and the effect of sample aging and to generate threshold density values useful for the detection of ovalbumin in samples from ancient works of art. The efficacy of the developed dot-blot immunoassay was proved testing microsamples from 13th–16th century mural paintings of Saint Francesco Church in Lodi (Italy). Despite the aging, the altered conditions of conservation, the complex matrix, and the micro-size of samples, the presence of ovalbumin was detected in all those mural painting samples where mass-spectrometry-based proteomic analysis unambiguously detected ovalbumin peptides

HEBE, a novel positive regulator of senescence in Solanum lycopersicum

Leaf senescence and plant aging are traits of great interest for breeders. Senescing cells undergo important physiological and biochemical changes, while cellular structures such as chloroplasts are degraded with dramatic metabolic consequences for the whole plant. The possibility of prolonging the photosynthetic ability of leaves could positively impact the plant\u2019s life span with benefits for biomass production and metabolite accumulation; plants with these characteristics display a stay-green phenotype. A group of plant transcription factors known as NAC play a pivotal role in controlling senescence: here we describe the involvement of the tomato NAC transcription factor Solyc12g036480, which transcript is present in leaves and floral buds. Since its silencing delays leaf senescence and prevents plants from ageing, we renamed Solyc12g0364 H\u1e16B\u112, for the Greek goddess of youth. In this manuscript we describe how HEB downregulation negatively affects the progression of senescence, resulting in changes in transcription of senescence-promoting genes, as well as the activity of enzymes involved in chlorophyll degradation, thereby explaining the stay-green phenotype

The 'Moon Mapping' project to promote cooperation between students of Italy and China

The research project 'Moon Mapping' has been established in 2014 between the Italian and Chinese Governments to promote cooperation and exchange between undergraduate students from both countries. The operational phase of the project started in early 2015, and will end in 2017, for a total length of three years. The main aim is to train new scholars to be able to work on different kinds of remotely-sensed data collected over the Moon surface by the Chinese space missions Chang'E-1/2. The project coordination has been assigned to the Italian Space Agency for the Italian side and to the Center of Space Exploration, China Ministry of Education, for the Chinese side. Several Chinese universities and Italian national research institutes and universities have been officially involved in this project. Six main research topics have been identified: (1) map of the solar wind ion; (2) geomorphological map of the Moon; (3) data preprocessing of Chang'E-1 mission; (4) map of element distribution; (5) establishment of 3D digital visualization system; and (6) compilation and publication of a tutorial on joint lunar mapping

PERFORMANCE EVALUATION OF A STRUCTURE AND MOTION STRATEGY IN ARCHITECTURE AND CULTURAL HERITAGE

A large number of strategies and implementations of Structure from Motion (SFM) have been developed; some are being included in photogrammetric software packages. However, finding specific parameters highlighting the efficiency, the accuracy and the reliability of a SFM strategy it is difficult. While most strategies will almost always succeed in any given task (i.e. the orientation of a certain image sequence), judging how good is actually the result, is somehow still an open issue. In the first part of this paper some alternatives and parameters are presented and their pro and cons discussed, with the objective to find out whether there are alternatives to the well-established method of the check points. In the second part, the results of most of the image sequences made available by the workshop organizers, processed with the SFM approach developed by the authors, are presented; results for the sequences where external information (such as ground control points or a DTM) were available are presented in more detail

Mobilization of sulfane sulfur from cysteine desulfurases to the Azotobacter vinelandii sulfurtransferase RhdA

Mobilization of the l-cysteine sulfur for the persulfuration of the rhodanese of Azotobacter vinelandii, RhdA, can be mediated by the A. vinelandii cysteine desulfurases, IscS and NifS. The amount of cysteine was higher in mutant strains lacking rhdA (MV474) than in wild type. The diazotrophic growth of MV474 was impaired. Taking into account the functional results about rhodanese-like proteins and RhdA itself, it is suggested that RhdA-dependent modulation of l-cysteine levels must deal with a redox-related process

Reinvestigation of the tautomerism of some substituted 2-hydroxypyridines

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Evaluation of perioperative complications using a newly described staging system for placenta accreta spectrum

Introduction: The antenatal diagnosis of placenta accreta spectrum (PAS) is in large part subjective and based on expert interpretation. The aim of this study was to externally evaluate a recently developed staging system based on specific and defined prenatal ultrasound (US) features in a cohort of women at risk of PAS undergoing specialist prenatal US, in particular relating to surgical morbidity at delivery. Materials and methods: Database study of cases with confirmed placenta previa. In all, the placenta was evaluated in a systematic fashion. PAS was subclassified in PAS0-PAS3 according to the loss of clear zone, placental lacunae, bladder wall interruption, uterovesical hypervascularity and increased vascularity in the parametrial region. Results: 43 cases were included, of whom 33 had major placenta previa. 31 cases were categorized as PAS0; 3, 4 and 5 cases as PAS1, PAS2 and PAS3, respectively. All women underwent caesarean section and hysterectomy was required in 10. The comparison of the perinatal outcomes among the PAS categories yielded greater operative time (50 (35–129) minutes for PAS0 vs 70 (48–120) for PAS1 vs 95 (60–150) for PAS2 vs 100 (87–180) for PAS3, p < 0.001) and estimated blood loss (800 (500–2500) mls for PAS0 vs 3500 (800–7500) for PAS1 vs 2850 (500–7500) for PAS2 vs 6000 (2500–11000) for PAS3, p < 0.001) for the highest PAS categories, which were also associated with a higher rate of hysterectomy (p < 0.001), blood transfusion (p = 0.002) and admission to ITU or HDU (p < 0.001) and longer postoperative admission of 3 (1–9) days for PAS0 vs 3 (2–12) for PAS1 vs 4.5 (3–6) for PAS2 vs 5 (3–22) for PAS3, p = 0.02. Conclusion: Perioperative complications are closely associated with PAS stage. This information is useful for counselling women and may be important in allocating staff and infrastructure resources at the time of delivery