Development of a Simple Method for Labeling and Identification of Protein Binders in Art

Easel paintings are assets with an important historic and cultural value. They usually possess a multi-tiered structure, composed of di erent layers some of which may present protein binders, making it important to identify these materials for restoration and conservation purposes. We propose the identification of di erent protein binders by a new fluorescent labeling method employing a coumarin based chromophore, C392STP (sodium(E/Z)-4-(4-(2-(6,7-dimethoxycoumarin-3-yl)vinyl)benzoyl)-2,3,5,6-tetrafluorobenzenesulfo-nate). The method was optimized using commercial proteins and was further tested on proteins extracted from hen’s egg yolk, white bovine milk, and rabbit skin glue. To model more realistic conditions, paint models of easel paintings were prepared. The paint models were made with hen’s egg yolk, white bovine milk, and rabbit skin glue, mixed with di erent pigments and submitted to artificial aging. Then the extracted proteins from the paint models were labeled with C392 which allowed a sensitive and selective identification by polyacrylamide gel electrophoresis (PAGE) of the di erent protein binders used. As a final test, three 19th century easel paintings, from the Italian painter Giorgio Marini, were analyzed. The results show the potential of the proposed method for the identification of protein binders present in easel paintings

Tympanoplasty:Restrospecüve review of result

O objetivo deste trabalho é determinar os resultados de timpanoplastias realizadas a doentes com otite media crónica. O estudo baseia-se na análise retrospetiva de resultados de 201 timpanoplasüas realizadas em doentes com diagnóstico de oüte media crónica, operados no serviço de Otorrinolarìngologia do Hospital Prof. Doutor Fernando Fonseca EPE, nos anos 2005-2010. Os doentes foram avaliados segundo o sexo, idade, antecedentes pessoais otorrinolaringológicos, tipo de perfuração, técnica cirúrgica, enxerto utilizado, audiograma pre e pós-operatório e follow- up pós-operatório. Verifi cou-se melhoria estrutural e funcional com encerramento de 80% das perfurações e diminuição do gap aero-ósseo (AO) medio de 4 frequências (0.5, 1,,2,4kH2\ de 22dB no pré-operatório para 13dB no pós-operatório, com 80% dos doentes a apresentarem melhoria funcional. Estes resultados confirmam a eficácia da técnica cirúrgica

Seasonality and Prevalence of Leishmania major Infection in Phlebotomus duboscqi Neveu-Lemaire from Two Neighboring Villages in Central Mali

Phlebotomus duboscqi is the principle vector of Leishmania major, the causative agent of cutaneous leishmaniasis (CL), in West Africa and is the suspected vector in Mali. Although found throughout the country the seasonality and infection prevalence of P. duboscqi has not been established in Mali. We conducted a three year study in two neighboring villages, Kemena and Sougoula, in Central Mali, an area with a leishmanin skin test positivity of up to 45%. During the first year, we evaluated the overall diversity of sand flies. Of 18,595 flies collected, 12,952 (69%) belonged to 12 species of Sergentomyia and 5,643 (31%) to two species of the genus Phlebotomus, P. duboscqi and P. rodhaini. Of those, P. duboscqi was the most abundant, representing 99% of the collected Phlebotomus species. P. duboscqi was the primary sand fly collected inside dwellings, mostly by resting site collection. The seasonality and infection prevalence of P. duboscqi was monitored over two consecutive years. P. dubsocqi were collected throughout the year. Using a quasi-Poisson model we observed a significant annual (year 1 to year 2), seasonal (monthly) and village effect (Kemena versus Sougoula) on the number of collected P. duboscqi. The significant seasonal effect of the quasi-Poisson model reflects two seasonal collection peaks in May-July and October-November. The infection status of pooled P. duboscqi females was determined by PCR. The infection prevalence of pooled females, estimated using the maximum likelihood estimate of prevalence, was 2.7% in Kemena and Sougoula. Based on the PCR product size, L. major was identified as the only species found in flies from the two villages. This was confirmed by sequence alignment of a subset of PCR products from infected flies to known Leishmania species, incriminating P. duboscqi as the vector of CL in Mali

Coarctation of the aorta and mild to moderate developmental delay in a child with a de novo deletion of chromosome 15(q21.1q22.2)

BACKGROUND: Deletion of 15q21q22 is a rare chromosomal anomaly. To date, there have been nine reports describing ten individuals with different segmental losses involving 15q21 and 15q22. Many of these individuals have common features of growth retardation, hypotonia and moderate to severe mental retardation. Congenital heart disease has been described in three individuals with interstitial deletion involving this region of chromosome 15. CASE PRESENTATION: We report a child with coarctation of the aorta, partial agenesis of corpus callosum and mild to moderate developmental delay, with a de novo deletion of 15q21.1q22.2, detected by the array Comparative Genomic Hybridization (CGH). We utilized chromosome 15-specific microarray-based CGH to define the chromosomal breakpoints in this patient. CONCLUSION: This is the first description of mapping of an interstitial deletion involving the chromosome 15q21q22 segment using the chromosome 15-specific array-CGH. The report also expands the spectrum of clinical phenotype associated with 15q21q22 deletion

Cancer Survivor Study (CASUS) on colorectal patients: longitudinal study on physical activity, fitness, nutrition, and its influences on quality of life, disease recurrence, and survival. Rationale and design

Purpose: Evidence suggests that being physically active in combination with a healthy diet contributes to diminish colorectal cancer risk. However, if this is true for colorectal cancer primary prevention, the same is not clear for its recurrence after colorectal cancer treatments. Data on cancer survival are scarce, and there is a need for greater attention on these survivors’ lifestyle behavior. This manuscript describes rationale and design of the Cancer Survival Study (CASUS) on colorectal patients, a longitudinal observational study with the aim of investigating how physical activity, physical fitness, and dietary intake are related with their quality of life, disease recurrence, and survival. Methods: The CASUS on colorectal patients is a longitudinal cohort study on colorectal survivors, aged 18 years or older, recruited 6, 12, and 24 months after surgery. Upon recruitment, patients fill in a battery of questionnaires about physical activity, dietary intake, and quality of life, donate blood samples, do physical fitness tests, and use an accelerometer during 7 days. Repeated analyses will be performed to assess changes over time in physical activity, physical fitness, dietary intake, and other factors in relation to recurrence and survival. Conclusions: Results will contribute to highlight the role of physical activity, physical fitness, and nutrition in the quality of life of colorectal cancer survivors, recurrence, and survival. This study will provide important information for policymakers on the potential benefits of future physical activity and nutritional interventions, which are inexpensive, as a way to improve general health of colorectal cancer survivors.Luisa Soares-Miranda is supported by the grant SFRH/BPD/76947/2011 funded by FCT (QREN-POPH-Type 4.1-Advanced Training, subsidized by the European Social Fund and national funds of MEC), PTDC/DES/099018/2008-FCT/FCOMP-01-0124-FEDER-009573, and the Research Centre in Physical Activity Health and Leisure is supported by FCT: UID/DTP/00617/2013. Carla Costa is supported by the grant SFRH/BPD/96196/2013, funded by FCT (QREN-POPH-Type 4.1-Advanced Training, subsidized by the European Social Fund and national funds of MEC)

Long-term TNT and DNT contamination: 1-D modeling of natural attenuation in the vadose zone: case study, Portugal

The vadose zone of a trinitrotoluene (TNT) and dinitrotoluene (DNT) contaminated site was investigated to assess the mobility of those explosives under natural conditions. Located in the left margin of the River Tejo Basin, Portugal, the site is located on unconsolidated sediments. Wastewaters associated with the 50-year explosives production were disposed in excavated ponds, from where water would infiltrate and pollute the unsaturated and saturated parts of the local aquifers. Two boreholes were drilled to 9 m depth in such a former waste pond to investigate the contaminant's fate in the vadose zone. Sediment samples were taken every 1-2 m for analysis of the polynitroaromatics (p-NACs) and organic volatile compounds, pH, organic carbon content, cation exchange capacity and grain size analysis. The main contaminant was TNT representing >70 % of the total p-NACs concentration that peaked approximately 7 mg/kg in one borehole, even if the median in both boreholes was of similar to 1 mg/kg. DNT was 4-30 % of the total p-NACs and nitrotoluene (NT), up to 5 %. No other (volatile) organic compound was detected. The predominance of TNT as the main contaminant implies that any natural mass reduction has been inefficient to clean the site. Several 1-D model simulations of p-NACs cleaning of the vadose zone under natural conditions indicated that the most probable scenario of combined advection and partitioning will only remove TNT after 10's of years, whereas DNT and NT will hardly be removed. Such low concentrations and long times for the p-NACs removal, suggest that by now those compounds have been washed-out to a level below standard limits

Promoter methylation and large intragenic rearrangements of DPYD are not implicated in severe toxicity to 5-fluorouracil-based chemotherapy in gastrointestinal cancer patients

<p>Abstract</p> <p>Background</p> <p>Severe toxicity to 5-fluorouracil (5-FU) based chemotherapy in gastrointestinal cancer has been associated with constitutional genetic alterations of the dihydropyrimidine dehydrogenase gene (<it>DPYD</it>).</p> <p>Methods</p> <p>In this study, we evaluated <it>DPYD </it>promoter methylation through quantitative methylation-specific PCR and screened <it>DPYD </it>for large intragenic rearrangements in peripheral blood from 45 patients with gastrointestinal cancers who developed severe 5-FU toxicity. <it>DPYD </it>promoter methylation was also assessed in tumor tissue from 29 patients</p> <p>Results</p> <p>Two cases with the IVS14+1G > A exon 14 skipping mutation (c.1905+1G > A), and one case carrying the 1845 G > T missense mutation (c.1845G > T) in the DPYD gene were identified. However, <it>DPYD </it>promoter methylation and large <it>DPYD </it>intragenic rearrangements were absent in all cases analyzed.</p> <p>Conclusions</p> <p>Our results indicate that <it>DPYD </it>promoter methylation and large intragenic rearrangements do not contribute significantly to the development of 5-FU severe toxicity in gastrointestinal cancer patients, supporting the need for additional studies on the mechanisms underlying genetic susceptibility to severe 5-FU toxicity.</p

Antioxidant intake among Brazilian adults - The Brazilian Osteoporosis Study (BRAZOS): a cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Antioxidant nutrient intake and the lesser formation of free radicals seem to contribute to chronic diseases. The aim of the present study was to evaluate the intake profile of the main dietary antioxidants in a representative sample of the adult Brazilian population and discuss the main consequences of a low intake of these micronutrients on overall health.</p> <p>Methods</p> <p>The sample comprised 2344 individuals aged 40 years or older from 150 cities and was based on a probabilistic sample from official data. The research was conducted through in-home interviews administered by a team trained for this purpose. Dietary intake information was obtained through 24-h recall. The Nutrition Data System for Research software program was used to analyze data on the intake of vitamins A, C and E, selenium and zinc, which was compared to Dietary Reference Intakes (DRIs). Differences in intake according to sex, anthropometrics, socioeconomic status and region were also evaluated. The SPSS statistical package (version 13) was used for the statistical analysis. P-values < 0.05 were considered significant.</p> <p>Results</p> <p>Higher proportions of low intake in relation to recommended values were found for vitamin E (99.7%), vitamin A (92.4%) and vitamin C (85.1%) in both genders. Intake variations were found between different regions, which may reflect cultural habits.</p> <p>Conclusion</p> <p>These results should lead to the development of public health policies that encourage educational strategies for improving the intake of micronutrients, which are essential to overall health and prevention of non-communicable diseases.</p

The proteome of neural stem cells from adult rat hippocampus

BACKGROUND: Hippocampal neural stem cells (HNSC) play an important role in cerebral plasticity in the adult brain and may contribute to tissue repair in neurological disease. To describe their biological potential with regard to plasticity, proliferation, or differentiation, it is important to know the cellular composition of their proteins, subsumed by the term proteome. RESULTS: Here, we present for the first time a proteomic database for HNSC isolated from the brains of adult rats and cultured for 10 weeks. Cytosolic proteins were extracted and subjected to two-dimensional gel electrophoresis followed by protein identification through mass spectrometry, database search, and gel matching. We could map about 1141 ± 209 (N = 5) protein spots for each gel, of which 266 could be identified. We could group the identified proteins into several functional categories including metabolism, protein folding, energy metabolism and cellular respiration, as well as cytoskeleton, Ca(2+ )signaling pathways, cell cycle regulation, proteasome and protein degradation. We also found proteins belonging to detoxification, neurotransmitter metabolism, intracellular signaling pathways, and regulation of DNA transcription and RNA processing. CONCLUSIONS: The HNSC proteome database is a useful inventory which will allow to specify changes in the cellular protein expression pattern due to specific activated or suppressed pathways during differentiation or proliferation of neural stem cells. Several proteins could be identified in the HNSC proteome which are related to differentiation and plasticity, indicating activated functional pathways. Moreover, we found a protein for which no expression has been described in brain cells before