Carpenter Syndrome: Extended RAB23 Mutation Spectrum and Analysis of Nonsense-mediated mRNA Decay

Carpenter syndrome, a rare autosomal recessive disorder characterized by a combination of craniosynostosis, polysyndactyly, obesity, and other congenital malformations, is caused by mutations in RAB23, encoding a member of the Rab-family of small GTPases. In 15 out of 16 families previously reported, the disease was caused by homozygosity for truncating mutations, and currently only a single missense mutation has been identified in a compound heterozygote. Here, we describe a further 8 independent families comprising 10 affected individuals with Carpenter syndrome, who were positive for mutations in RAB23. We report the first homozygous missense mutation and in-frame deletion, highlighting key residues for RAB23 function, as well as the first splice-site mutation. Multi-suture craniosynostosis and polysyndactyly have been present in all patients described to date, and abnormal external genitalia have been universal in boys. High birth weight was not evident in the current group of patients, but further evidence for laterality defects is reported. No genotype-phenotype correlations are apparent. We provide experimental evidence that transcripts encoding truncating mutations are subject to nonsense-mediated decay, and that this plays an important role in the pathogenesis of many RAB23 mutations. These observations refine the phenotypic spectrum of Carpenter syndrome and offer new insights into molecular pathogenesis. (C) 2011 Wiley-Liss, Inc.

Population genetic analysis of large sequence polymorphisms in Plasmodium falciparum blood-stage antigens

Plasmodium falciparum, the causative agent of human malaria, invades host erythrocytes using several proteins on the surface of the invasive merozoite, which have been proposed as potential vaccine candidates. Members of the multi-gene PfRh family are surface antigens that have been shown to play a central role in directing merozoites to alternative erythrocyte receptors for invasion. Recently, we identified a large structural polymorphism, a 0.58 Kb deletion, in the C-terminal region of the PfRh2b gene, present at a high frequency in parasite populations from Senegal. We hypothesize that this region is a target of humoral immunity. Here, by analyzing 371 P. falciparum isolates we show that this major allele is present at varying frequencies in different populations within Senegal, Africa, and throughout the world. For allelic dimorphisms in the asexual stage antigens, Msp-2 and EBA-175, we find minimal geographic differentiation among parasite populations from Senegal and other African localities, suggesting extensive gene flow among these populations and/or immune-mediated frequency-dependent balancing selection. In contrast, we observe a higher level of inter-population divergence (as measured by F(st)) for the PfRh2b deletion, similar to that observed for SNPs from the sexual stage Pfs45/48 loci, which is postulated to be under directional selection. We confirm that the region containing the PfRh2b polymorphism is a target of humoral immune responses by demonstrating antibody reactivity of endemic sera. Our analysis of inter-population divergence suggests that in contrast to the large allelic dimorphisms in EBA-175 and Msp-2, the presence or absence of the large PfRh2b deletion may not elicit frequency-dependent immune selection, but may be under positive immune selection, having important implications for the development of these proteins as vaccine candidates. (C) 2009 Elsevier B.V. All rights reserved.

Shape classification using complex network and Multi-scale Fractal Dimension

Shape provides one of the most relevant information about an object. This makes shape one of the most important visual attributes used to characterize objects. This paper introduces a novel approach for shape characterization, which combines modeling shape into a complex network and the analysis of its complexity in a dynamic evolution context. Descriptors computed through this approach show to be efficient in shape characterization, incorporating many characteristics, such as scale and rotation invariant. Experiments using two different shape databases (an artificial shapes database and a leaf shape database) are presented in order to evaluate the method. and its results are compared to traditional shape analysis methods found in literature. (C) 2009 Published by Elsevier B.V.

Analysis of the combined effect of lasers of different wavelengths for PDT outcome using 600, 630, and 660 nm

We investigated the effects of photodynamic therapy (PDT) outcome when combining three laser systems that produce light in three different wavelengths (600, 630, and 660 nm). Cooperative as well as independent effects can be observed. We compared the results of the combined wavelengths of light with the effect of single laser for the excitation of the photosensitizer. In the current experiment, the used photosensitizer was Photogem (R) (1.5 mg/kg). Combining two wavelengths for PDT, their cumulative dose and different penetrability may change the overall effect of the fluence of light, which can be effective for increasing the depth of necrosis. This evaluation was performed by comparing the depth and specific aspect of necrosis obtained by using single and dual wavelengths for irradiation of healthy liver of male Wistar rats. We used 15 animals and divided them in five groups of three animals. First, Photogem (R) was administered; follow by measurement of the fluorescence spectrum of the liver before PDT to confirm the level of accumulation of photosensitizer in the tissue. After that, an area of 1 cm(2) of the liver was illuminated using different laser combinations. Qualitative analysis of the necrosis was carried out through histological and morphological study. [GRAPHICS] (a) - microscopic images of rat liver cells, (b) - superficial necrosis caused by PDT using dual-wavelength illumination, (c) - neutrophilic infiltration around the vessel inside the necrosis, and (d) - neutrophilic infiltration around the vessel between necrosis and live tissue (C) 2011 by Astro Ltd. Published exclusively by WILEY-VCH Verlag GmbH & Co. KGaA

Nickel hydroxide electrodes as amperometric detectors for carbohydrates in flow injection analysis and liquid chromatography

The present paper describes the utilization of nickel hydroxide modified electrodes toward the catalytic oxidation of carbohydrates (glucose, fructose, lactose and sucrose) and their utilization as electrochemical sensor. The modified electrodes were employed as a detector in flow injection analysis for individual carbohydrate detection, and to an ionic column chromatography system for multi-analyte samples aiming a prior separation step. Kinetic studies were performed on a rotating disk electrode (RDE) in order to determine both the heterogeneous rate constant and number of electrons transferred for each carbohydrate. Many advantages were found for the proposed system including fast and easy handling of the electrode modification, low cost procedure, a wide range of linearity (0.5-50 ppm), low detection limits (ppb level) and high sensitivities. (C) 2009 Elsevier B.V. All rights reserved.

Elemental analysis of nuts and seeds by axially viewed ICP OES

A method for the multi-elemental determination of metals (Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Sr and Zn), metalloids (B and Si), and non-metals (Cl, P and 5) in the babassu nut and mesocarp, sapucaia nut, coconut pulp, cupuassu pulp and seed, and cashew nut by axially viewed inductively coupled plasma optical emission spectrometry is presented. A diluted oxidant mixture (2 ml HNO(3) + 1 ml H(2)O(2) + 3 ml H(2)O) was used to achieve the complete decomposition of the organic matrix in a closed-vessel microwave oven. The accuracy of the entire proposed method was confirmed by standard reference material analysis (peach leaves-NIST SRM1547). The certified values showed a good agreement at a 95% confidence limit (Student`s t-test). The average RSD for repeatability of calibration solutions measurements were in the range of 1.1-6.7%. Limits of quantification (LOQ = 10 x LOD) were in the level of 0.00072-0.0532 mg/l. The macro and micronutrient ranges in the different nuts and seeds did not exceed the dietary reference intake (DRI), except for Mn in the babassu nut. (C) 2010 Published by Elsevier Ltd.

Automated two-dimensional separation flow system with electrochemical preconcentration, stripping, capillary electrophoresis and contactless conductivity detection for trace metal ion analysis

This paper describes the automation of a fully electrochemical system for preconcentration, cleanup, separation and detection, comprising the hyphenation of a thin layer electrochemical flow cell with CE coupled with contactless conductivity detection (CE-C(4)D). Traces of heavy metal ions were extracted from the pulsed-flowing sample and accumulated on a glassy carbon working electrode by electroreduction for some minutes. Anodic stripping of the accumulated metals was synchronized with hydrodynamic injection into the capillary. The effect of the angle of the slant polished tip of the CE capillary and its orientation against the working electrode in the electrochemical preconcentration (EPC) flow cell and of the accumulation time were studied, aiming at maximum CE-C(4)D signal enhancement. After 6 min of EPC, enhancement factors close to 50 times were obtained for thallium, lead, cadmium and copper ions, and about 16 for zinc ions. Limits of detection below 25 nmol/L were estimated for all target analytes but zinc. A second separation dimension was added to the CE separation capabilities by staircase scanning of the potentiostatic deposition and/or stripping potentials of metal ions, as implemented with the EPC-CE-C(4)D flow system. A matrix exchange between the deposition and stripping steps, highly valuable for sample cleanup, can be straightforwardly programmed with the multi-pumping flow management system. The automated simultaneous determination of the traces of five accumulable heavy metals together with four non-accumulated alkaline and alkaline earth metals in a single run was demonstrated, to highlight the potentiality of the system.

En jämförelse av molnbaserade realtidsanalystjänster : Skillnader i användbarhet mellan multi-tenant och single-tenant arkitekturer

Internet of Things är ett samlingsbegrepp för den utveckling som innebär att olika typer av enheter kan förses med sensorer och datachip som är uppkopplade mot internet. En ökad mängd data innebär en ökad förfrågan på lösningar som kan lagra, spåra, analysera och bearbeta data. Ett sätt att möta denna förfrågan är att använda sig av molnbaserade realtidsanalystjänster. Multi-tenant och single-tenant är två typer av arkitekturer för molnbaserade realtidsanalystjänster som kan användas för att lösa problemen med hanteringen av de ökade datamängderna. Dessa arkitekturer skiljer sig åt när det gäller komplexitet i utvecklingen. I detta arbete representerar Azure Stream Analytics en multi-tenant arkitektur och HDInsight/Storm representerar en single-tenant arkitektur. För att kunna göra en jämförelse av molnbaserade realtidsanalystjänster med olika arkitekturer, har vi valt att använda oss av användbarhetskriterierna: effektivitet, ändamålsenlighet och användarnöjdhet. Vi kom fram till att vi ville ha svar på följande frågor relaterade till ovannämnda tre användbarhetskriterier: • Vilka likheter och skillnader kan vi se i utvecklingstider? • Kan vi identifiera skillnader i funktionalitet? • Hur upplever utvecklare de olika analystjänsterna? Vi har använt en design and creation strategi för att utveckla två Proof of Concept prototyper och samlat in data genom att använda flera datainsamlingsmetoder. Proof of Concept prototyperna inkluderade två artefakter, en för Azure Stream Analytics och en för HDInsight/Storm. Vi utvärderade dessa genom att utföra fem olika scenarier som var för sig hade 2-5 delmål. Vi simulerade strömmande data genom att låta en applikation kontinuerligt slumpa fram data som vi analyserade med hjälp av de två realtidsanalystjänsterna. Vi har använt oss av observationer för att dokumentera hur vi arbetade med utvecklingen av analystjänsterna samt för att mäta utvecklingstider och identifiera skillnader i funktionalitet. Vi har även använt oss av frågeformulär för att ta reda på vad användare tyckte om analystjänsterna. Vi kom fram till att Azure Stream Analytics initialt var mer användbart än HDInsight/Storm men att skillnaderna minskade efter hand. Azure Stream Analytics var lättare att arbeta med vid simplare analyser medan HDInsight/Storm hade ett bredare val av funktionalitet.

Women's acceptability of misoprostol treatment for incomplete abortion by midwives and physicians : secondary outcome analysis from a randomized controlled equivalence trial at district level in Uganda

OBJECTIVE: This study aimed to assess women´s acceptability of diagnosis and treatment of incomplete abortion with misoprostol by midwives, compared with physicians. METHODS: This was an analysis of secondary outcomes from a multi-centre randomized controlled equivalence trial at district level in Uganda. Women with first trimester incomplete abortion were randomly allocated to clinical assessment and treatment with misoprostol by a physician or a midwife. The randomisation (1:1) was done in blocks of 12 and stratified for health care facility. Acceptability was measured in expectations and satisfaction at a follow up visit 14-28 days following treatment. Analysis of women's overall acceptability was done using a generalized linear mixed-effects model with an equivalence range of -4% to 4%. The study was not masked. The trial is registered at ClinicalTrials.org, NCT 01844024. RESULTS: From April 2013 to June 2014, 1108 women were assessed for eligibility of which 1010 were randomized (506 to midwife and 504 to physician). 953 women were successfully followed up and included in the acceptability analysis. 95% (904) of the participants found the treatment satisfactory and overall acceptability was found to be equivalent between the two study groups. Treatment failure, not feeling calm and safe following treatment, experiencing severe abdominal pain or heavy bleeding following treatment, were significantly associated with non-satisfaction. No serious adverse events were recorded. CONCLUSIONS: Treatment of incomplete abortion with misoprostol by midwives and physician was highly, and equally, acceptable to women. TRIAL REGISTRATION: ClinicalTrials.gov NCT01844024.

Purification and analysis of protein synthetic initiation factors from Volvox Carteri

Volvox carteri, a multi-celled green algae, can grow synchronously given a sixteen hour light period followed by an eight hour dark period, a cycle which is repeated for a 48 hour growth cycle total. Near the end of each light period, reproductive cells divide rapidly resulting in the differentiation of ceIls. When the dark period begins, this differentiation stops and the cells remain dormant with little protein synthesis or differentiation occurring. Immediately after the lights come back on, however, the cells again undergo rapid protein synthesis and complete their differentiation. Previous studies have concluded that Volvox carteri discontinue protein synthesis during the dark phase due to regulation at the translational level and not the transcriptional level. Therefore, the inhibition of protein synthesis does not lie in the transfer of the protein coding sequence from DNA to mRNA, but rather in the transfer of this information from the mRNA to the ribosomes. My research examined this translational regulation to determine the factor(s) causing the discontinuation of protein synthesis during the dark phase. Evidence from other research further suggests that the control of translation lies in the initiation step rather than the elongation step. Eukaryotic initiation factors aid in the binding of the ribosomal subunits to the mRNA to initiate protein synthesis. It is known that initiation factors can be modified by phosphorylation, regulating their activity. Therefore, my study focused upon isolating some of these initiation factors in order to determine whether or not such modifications are responsible for the inhibition of dark phase protein synthesis in Volvox carteri.

An Environmental Analysis Corroborating PDA and the Winthrop Example

Libraries seek active ways to innovate amidst macroeconomic shifts, growing online education to help alleviate ever-growing schedule conflicts as students juggle jobs and course schedules, as well as changing business models in publishing and evolving information technologies. Patron-driven acquisition (PDA), also known as demand-driven acquisition (DDA), offers numerous strengths in supporting university curricula in the context of these significant shifts. PDA is a business model centered on short-term loans and subsequent purchases of ebooks resulting directly from patrons' natural use stemming from their discovery of the ebooks in library catalogs where the ebooks' bibliographic records are loaded at regular intervals established between the library and ebook supplier. Winthrop University's PDA plan went live in October 2011, and this article chronicles the philosophical and operational considerations, the in-library collaboration, and technical preparations in concert with the library system vendor and ebook supplier. Short-term loan is invoked after a threshold is crossed, typically number of pages or time spent in the ebook. After a certain number of short-term loans negotiated between the library and ebook supplier, the next short-term loan becomes an automatic purchase after which the library owns the ebook in perpetuity. Purchasing options include single-user and multi-user licenses. Owing to high levels of need in college and university environments, Winthrop chose the multi-user license as the preferred default purchase. Only where multi-user licenses are unavailable does the automatic purchase occur with single-user title licenses. Data on initial use between October 2011 and February 2013 reveal that of all PDA ebooks viewed, only 30% crossed the threshold into short-term loans. Of all triggered short-term loans, Psychology was the highest-using. Of all ebook views too brief to trigger short-term loans, Business was the highest-using area. Although the data are still too young to draw conclusions after only a few months, thought-provoking usage differences between academic disciplines have begun to emerge. These differences should be considered in library plans for the best possible curricular support for each academic program. As higher education struggles with costs and course-delivery methods libraries have an enduring lead role.

Performance analysis and analysis for performance : a study of Villa-Lobos's Bachianas Brasileiras No. 9

This study of Villa-Lobos’s Bachianas Brasileiras No. 9 aims at gaining insight into the decision-making processes of translating a score into a musical performance. Chapter I presents a discussion of selected issues related to interpretative analysis. Chapter II is an overview of the approaches to recording comparison deemed relevant to the present study. Chapter III is a comparative study of the vocal and string versions of Bachianas Brasileiras No. 9, while Chapter IV offers a structural analysis of the work. Chapter V compares four recordings: the composer’s own with the Orchestre National de La Radiodiffusion Française—EMI 7243 5 66964 2 6; Odaline de la Martinez and the BBC Singers, LNT 102; Michael Tilson Thomas and the New World Symphony —RCA 09026-68538-2; and my own CD, Construção, Orquestra de Câmara Theatro São Pedro- Limited Edition (live recording made on December 11, 1995 in Bayreuth, Germany). This comparison utilizes data obtained with the software Tempo. The tabulation of these results is shown in graphs that compare how matters of tempo flexibility affect each performance. This multi-faceted study shows that although painstaking analysis can lead to insightful solutions, the fleeting nature of musical performance requires an open mind and imagination to deal with the often contradictory directives of the score.