Increased clopidogrel response is associated with ABCC3 expression: A pilot study

Background: The aim of this study was investigate the relationship between ABCB1 and ABCC3 gene expressions in peripheral blood cells (PBC) and the response to clopidogrel in patients with coronary arterial disease (CAD). Methods: Twenty-six male CAD patients (50-70 years) under treatment with clopidogrel (75 mg/day) for at least 5 days were selected. Blood samples were obtained to evaluate platelet reactivity and ABCB1 and ABCC3 mRNA expression. Platelet reactivity was measured in P2Y12 Reaction Units (PRU) using VerifyNow. RNA was extracted from PBC and mRNA levels were measured by qPCR, using GAPD as a reference gene. Results: Platelet response to clopidogrel was categorized in to PRU quartiles. Individuals with PRU values within the first quartile (Q1, <151 units) were considered good responders, while those who had PRU within the fourth quartile (Q4. PRU>260) were considered non-responders. ABCC3 was 1.7 times more expressed in Q4 than in Q1 PRU group (p=0.048). Moreover, CAD patients with low ABCC3 expression (Qe1, <2.5x10(-3)) had higher probability to have a good response to clopidogrel (OR: 18.00, 95%CI: 1.90-169.99, p=0.001). Univariate linear regression analysis demonstrated that low ABCC3 mRNA expression contributed with a reduction of 73 PRU in relation to the patients with expression value higher than 2.5x10(-3) (p=0.027). Neither ABCB1 mRNA levels nor clinical variables studied influenced PRU values. Conclusions: Low ABCC3 mRNA expression in peripheral blood cells is associated with increased clopidogrel response, but further studies are needed to describe the functional relationship of clopidogrel with the ABCC3. Crown Copyright (C) 2011 Published by Elsevier B.V. All rights reserved.

Lipid transfers to HDL are predictors of precocious clinical coronary heart disease

Background: High-density-lipoprotein (HDL) has several antiatherogenic properties and, although the concentration of HDL-cholesterol negatively correlates with incidence of coronary artery disease (CAD), this is not sufficient to evaluate the overall HDL protective role. The aim was to investigate whether precocious CAD patients show abnormalities in lipid transfers to HDL, a fundamental step in HDL metabolism and function. Methods: Thirty normocholesterolemic CAD patients aged <50 y and 30 controls paired for sex, age and B.M.I. were studied. Fasting blood samples were collected for the in vitro lipid transfer assay and plasma lipid determination. A donor nanoemulsion labeled with radioactive free-cholesterol. cholesteryl esters, phospholipids and triglycerides was incubated with whole plasma and after chemical precipitation of non-HDL fractions, supernatant was counted for radioactivity in HDL. Results: LDL and HDL-cholesterol and triglycerides were equal in both groups. Transfers of free-cholesterol (3.8 +/- 1.2%vs 7.0 +/- 33%,p<0.0001) and triglycerides (3.7 +/- 1.7%vs 4.9 +/- 1.9%, p = 0.0125) were diminished in CAD patients whereas cholesteryl ester transfer increased (6.5 +/- 1.9%vs 4.8 +/- 1.8%, p = 0.0008); phospholipid transfer was equal (17.8 +/- 3.5% vs19.5 +/- 3.9%). Conclusion: Alterations in the transfer of lipids to HDL may constitute a new marker for precocious CAD and relation of this metabolic alteration with HDL antiatherogenic function should be investigated in future studies. (C) 2011 Published by Elsevier B.V.

The importance of landscape structure for seed dispersal in rain forest fragments

Questions What are the main features of the seed rain in a fragmented Atlantic forest landscape? Can seed rain species attributes (life form, dispersal mode, successional status) relate to the spatial arrangement (size and number of fragments, edge density and presence of corridor) of forest fragments in the landscape? How does the rain forest landscape structure affect the seed rain? Location Atlantic rainforest, Sao Paulo State, Southeastern Brazil. Methods Seed rain samples were collected monthly throughout 1yr, counted, identified and classified according to species dispersal mode, successional status and life form. Seed rain composition was compared with woody species near the seed traps. Relationships between seed rain composition and landscape spatial arrangement (fragment area, presence of corridor, number of fragments in the surroundings, proximity of fragments, and edge density) were tested using canonical correspondence analysis (CCA). Results We collected 20142 seeds belonging to 115 taxa, most of them early successional and anemochorous trees. In general, the seed rain had a species composition distinct from that of the nearby forest tree community. Small isolated fragments contained more seeds, mainly of anemochorous, epiphytic and early-successional species; large fragments showed higher association with zoochorous and late-successional species compared to small fragments. The CCA significantly distinguished the species dispersal mode according to fragment size and isolation, anemochorous species being associated to small and isolated fragments, and zoochorous species to larger areas and fragment aggregation. Nevertheless, a gradient driven by proximity (PROX) and edge density (ED) segregated lianas (in the positive extremity), early successional and epiphyte species (in the negative end); large fragments were positively associated to PROX and ED. Conclusions The results highlight the importance of the size and spatial arrangement of forest patches to promote habitat connectivity and improve the flux of animal-dispersed seeds. Landscape structure controls seed fluxes and affects plant dispersal capacity, potentially influencing the composition and structure of forest fragments. The seed rain composition may be used to assess the effects of landscape spatial structure on plant assemblages, and provide relevant information for biodiversity conservation.

Aplicação do método de ensaio das frequências naturais de vibração para obtenção do módulo de elasticidade de peças estruturais de madeira

Existem diversas técnicas para caracterização do módulo de elasticidade de madeiras e, dentre as atualmente empregadas, destacam-se aquelas que utilizam as frequências naturais de vibração, por serem técnicas não destrutivas e, portanto, apresentarem resultados que podem ser repetidos e comparados ao longo do tempo. Este trabalho teve como objetivo avaliar a eficácia, dos métodos de ensaios baseados nas frequências naturais de vibração comparando-os aos resultados obtidos na flexão estática na obtenção das propriedades elásticas em peças estruturais de madeira de reflorestamento que são usualmente empregadas na construção civil. Foram avaliadas 24 vigas de Eucalyptus sp. com dimensões nominais (40 x 60 x 2.000 mm) e 14 vigas de Pinus oocarpa com dimensões nominais (45 x 90 x 2.300 mm), ambas sem tratamento; 30 pranchas com dimensões nominais (40 x 240 x 2.010 mm) e 30 pranchas com dimensões nominais (40 x 240 x 3.050 mm), ambas de Pinnus oocarpa e com tratamento preservativo à base de Arseniato de Cobre Cromatado - CCA. Os resultados obtidos apresentaram boa correlação quando comparados aos resultados obtidos pelo método mecânico de flexão estática, especialmente quando empregada a frequência natural de vibração longitudinal. O emprego da frequência longitudinal mostrou-se confiável e prático, portanto recomendada para a determinação do módulo de elasticidade de peças estruturais de madeira. Verificou-se ainda que, empregando a frequência longitudinal, não há necessidade de um suporte específico para os corpos de prova ou calibrações prévias, reduzindo assim o tempo de execução e favorecendo o ensaio de grande quantidade de amostras.

Os sentidos da silicose atribuídos por trabalhadores de pedreiras adoecidos

Este estudo teve o objetivo de identificar os sentidos atribuídos à silicose entre trabalhadores de pedreiras adoecidos. Foi baseado na antropologia médica e na história de vida focal. Os dados foram coletados por entrevistas e observações diretas realizadas no domicílio de oito portadores de silicose, cadastrados no Serviço de Vigilância Epidemiológica de Guapé - Minas Gerais. A técnica de análise temática indutiva foi usada para sintetizar os núcleos temáticos: a explicação para a doença, a sua gravidade e consequências. Segundo os trabalhadores, a doença é permeada pelo sofrimento e alteração da identidade social, contextualizada no trabalho de homens jovens com manuseio da sílica. Esses sentidos fornecem uma base com a qual o enfermeiro do trabalho pode prestar um cuidado diferenciado, desenvolver ações de prevenção aos riscos à saúde, de adequação do ambiente de trabalho, além do aperfeiçoamento das estratégias de notificação dos agravos e de adesão ao tratamento.

Pozolanicidade dos resíduos industriais, lã de vidro e lã cerâmica

As fibras cerâmicas se caracterizam por ser um material leve, com alto grau de pureza, baixo armazenamento de calor, baixa condutividade térmica, resistência a choque térmico e alta resistência à corrosão em altas temperaturas. Essas características levam a uma grande procura das indústrias mínero-metalúrgicas e de outros setores para revestimentos de distribuidores, muflas, fornos de aquecimentos, entre outros. Após utilização no processo, por perderem sua capacidade de isolamento, os resíduos gerados precisam de destinação. Esse trabalho enfoca, especificamente, resíduos de lã cerâmica e lã de vidro. Pelo fato de a composição das fibras cerâmicas ser rica em sílica e alumina, efetuou-se uma investigação acerca da atividade pozolânica das mesmas com a cal e o cimento, especificamente CPV ARI, CPII E32 e CPIII 32RS, para avaliação da perspectiva de reciclagem em possível incorporação no concreto.

Relationship between space distribution of the benthic macroinvertebrates community and trophic state in a Neotropical reservoir (Itupararanga, Brazil)

The purpose of this work was to verify the benthic macroinvertebrates community responses through environmental factors along a headwater tropical reservoir. Samplings were taken with a Van-Veen grab along the reservoir in littoral and profundal regions and in the headwater, next to the dam and the middle of the reservoir. Samples were taken during both wet and dry seasons. Dissolved oxygen concentrations, electric conductivity, temperature and pH near the sediment have been performed in situ, at every sampling station by using a multiprobe and Secchi disc. Total water phosphorus and chlorophyll a concentrations were analyzed to determine the trophic state index. Sediment's organic matter, total phosphorus, nitrogen concentrations and granulometric composition were measured. In order to verify which environmental variables would have more influence over the benthic macroinvertebrates community, a canonical correspondence analysis (CCA) was performed. The total number of recorded taxa was 28. Among them, the family Chironomidae (Diptera) was the richest group (19 taxa). It can be proposed that the benthic macroinvertebrates community may be influenced by environmental conditions such as nutrient and organic matter availability, as well as dissolved oxygen concentration. Macroinvertebrates are adequate bioindicators of water quality due to their sensibility to environmental changes mentioned before. Chironomus sp, Limnodrilus hoffmeisteri and Branchiura sowerbyi comprises a group that can be considered bio-indicators of eutrophic conditions. A second group can be considered as indicator of mesotrophic conditions. The presence of two or more members from that group which comprises Tanytarsini spp, Fissimentum sp, Pelomus sp and Goeldichironomus sp, like predominant taxa, may indicates mesotrophic conditions.

Estudio hidrogeológico de los barrancos de Guiniguada y Tenoya (Gran Canaria)

[ES] El sector Noreste de la Isla de Gran Canaria muestra un sistema de flujo del agua subterránea de cumbre a costa, con desviaciones hacia el fondo del profundo barranco Guiniguada, donde posiblemente estaban los antiguos nacientes, hoy captados por pozos. Existen en la zona 234 captaciones, fundamentalmente pozos de gran diámetro, de los que 140 están en uso, explotándose la Fm. Sálica a cotas bajas y los materiales del Grupo Roque Nublo a cotas elevadas. La salinidad crece de cumbre a costa por mayor efecto del aerosol marino y menor recarga. La recarga media puede variar desde menos de 20 mm/a en zonas bajas hasta más de 500 mm/a en cumbres, lo que puede suponer de 30 a 50 hm3/año que fluyen por la base de las formaciones volcánicas recientes y el techo de las mas antiguas. Los elevados contenidos en nitrato son atribuibles a la gran superficie cultivada. Cabe que la contaminación continúe creciendo pues parte del NO3 puede estar aún avanzado verticalmente por la gruesa zona no saturada.

Inibizione selettiva del gene MYCN mediante PNA (acidi peptido nucleici) anti-gene nel rabdomiosarcoma umano

MYCN oncogene amplification/expression is a feature of many childhood tumors, and some adult tumors, and it is associated with poor prognosis. While MYC expression is ubiquitary, MYCN has a restricted expression after birth and it is an ideal target for an effective therapy. PNAs belong to the latest class of nucleic acid-based therapeutics, and they can bind chromosomal DNA and block gene transcription (anti-gene activity). We have developed an anti-gene PNA that targets specifically the MYCN gene to block its transcription. We report for the first time MYCN targeted inhibition in Rhabdomyosarcoma (RMS) by the anti-MYCN-PNA in RMS cell lines (four ARMS and four ERMS) and in a xenograft RMS mouse model. Rhabdomyosarcoma is the most common pediatric soft-tissue sarcoma, comprising two main subgroups [Alveolar (ARMS) and Embryonal (ERMS)]. ARMS is associated with a poorer prognosis. MYCN amplification is a feature of both the ERMS and ARMS, but the MYCN amplification and expression levels shows a significant correlation and are greater in ARMS, in which they are associated with adverse outcome. We found that MYCN mRNA and protein levels were higher in the four ARMS (RH30, RH4, RH28 and RMZ-RC2) than in the four ERMS (RH36, SMS-CTR, CCA and RD) cell lines. The potent inhibition of MYCN transcription was highly specific, it did not affect the MYC expression, it was followed by cell-growth inhibition in the RMS cell lines which correlated with the MYCN expression rate, and it led to complete cell-growth inhibition in ARMS cells. We used a mutated- PNA as control. MYCN silencing induced apoptosis. Global gene expression analysis (Affymetrix microarrays) in ARMS cells treated with the anti-MYCN-PNA revealed genes specifically induced or repressed, with both genes previously described as targets of N-myc or Myc, and new genes undescribed as targets of N-myc or Myc (mainly involved in cell cycle, apoptosis, cell motility, metastasis, angiogenesis and muscle development). The changes in the expression of the most relevant genes were confirmed by Real-Time PCR and western blot, and their expression after the MYCN silencing was evaluated in the other RMS cell lines. The in vivo study, using an ARMS xenograft murine model evaluated by micro-PET, showed a complete elimination of the metabolic tumor signal in most of the cases (70%) after anti-MYCN-PNA treatment (without toxicity), whereas treatment with the mutated-PNA had no effect. Our results strongly support the development of MYCN anti-gene therapy for the treatment of RMS, particularly for poor prognosis ARMS, and of other MYCN-expressing tumors.

Intra-arterial injection of neural stem cells using a microneedle technique does not cause microembolic strokes

Intra-arterial (IA) injection represents an experimental avenue for minimally invasive delivery of stem cells to the injured brain. It has however been reported that IA injection of stem cells carries the risk of reduction in cerebral blood flow (CBF) and microstrokes. Here we evaluate the safety of IA neural progenitor cell (NPC) delivery to the brain. Cerebral blood flow of rats was monitored during IA injection of single cell suspensions of NPCs after stroke. Animals received 1 × 10(6) NPCs either injected via a microneedle (microneedle group) into the patent common carotid artery (CCA) or via a catheter into the proximally ligated CCA (catheter group). Controls included saline-only injections and cell injections into non-stroked sham animals. Cerebral blood flow in the microneedle group remained at baseline, whereas in the catheter group a persistent (15 minutes) decrease to 78% of baseline occurred (P<0.001). In non-stroked controls, NPCs injected via the catheter method resulted in higher levels of Iba-1-positive inflammatory cells (P=0.003), higher numbers of degenerating neurons as seen in Fluoro-Jade C staining (P<0.0001) and ischemic changes on diffusion weighted imaging. With an appropriate technique, reduction in CBF and microstrokes do not occur with IA transplantation of NPCs.

Percutaneous and Open Retrograde Endovascular Stenting of Symptomatic High-Grade Innominate Artery Stenosis: Technique and Follow-Up

Angioplasty and stenting of the IA have been reported with high technical and clinical success rates, low complication rates and good mid-term patency rates. Different antegrade or retrograde endovascular catheter-based approaches and combinations with surgical exposure of the CCA are used. The purpose of this study was to determine safety, efficacy and mid-term clinical and radiological outcome of the stent-assisted treatment of atherosclerotic stenotic disease of the IA with special focus on the different technical approaches.

Circulation dynamics and its influence on European and Mediterranean January-April climate over the past half millennium: results and insights from instrumental data, documentary evidence and coupled climate models

We use long instrumental temperature series together with available field reconstructions of sea-level pressure (SLP) and three-dimensional climate model simulations to analyze relations between temperature anomalies and atmospheric circulation patterns over much of Europe and the Mediterranean for the late winter/early spring (January–April, JFMA) season. A Canonical Correlation Analysis (CCA) investigates interannual to interdecadal covariability between a new gridded SLP field reconstruction and seven long instrumental temperature series covering the past 250 years. We then present and discuss prominent atmospheric circulation patterns related to anomalous warm and cold JFMA conditions within different European areas spanning the period 1760–2007. Next, using a data assimilation technique, we link gridded SLP data with a climate model (EC-Bilt-Clio) for a better dynamical understanding of the relationship between large scale circulation and European climate. We thus present an alternative approach to reconstruct climate for the pre-instrumental period based on the assimilated model simulations. Furthermore, we present an independent method to extend the dynamic circulation analysis for anomalously cold European JFMA conditions back to the sixteenth century. To this end, we use documentary records that are spatially representative for the long instrumental records and derive, through modern analogs, large-scale SLP, surface temperature and precipitation fields. The skill of the analog method is tested in the virtual world of two three-dimensional climate simulations (ECHO-G and HadCM3). This endeavor offers new possibilities to both constrain climate model into a reconstruction mode (through the assimilation approach) and to better asses documentary data in a quantitative way.

A new DPYD genotyping assay for improving the safety of 5-fluorouracil therapy

Chemotherapeutic use of 5-fluorouracil (5FU) is compromised by 10-20% of patients developing severe toxicity. Recently described genetic variation in dihydropyrimidine dehydrogenase (DPYD) has been shown to be a major predictor of 5FU toxicity. Here, we describe a new genotyping assay for routine clinical use that covers all the major DPYD risk variants.

Relationship between glomerular filtration rate and the adipokines adiponectin, resistin and leptin in coronary patients with predominantly normal or mildly impaired renal function

BACKGROUND: Due to their molecular weight, it is possible that the adipokines adiponectin, resistin and leptin accumulate when glomerular filtration rate (GFR) is decreased. In reduced renal clearance, altered serum concentrations of these proteins might affect cardiovascular risk. The objective of the study was to investigate the relationship between adipokine concentrations and GFR. METHODS: The association between GFR, as determined by the abbreviated MDRD equation, and the concentrations of the adipokines adiponectin, resistin and leptin was assessed in a cohort of coronary patients (n=538; 363 male, 165 female). After calculation of correlations between GFR and adipokine concentrations, the association was further assessed by analysis of covariance following adjustment for age, gender, BMI, presence of type 2 diabetes, presence of hypertension, history of smoking as well as for serum lipid concentrations. RESULTS: Mean GFR in our study population was 68.74+/-15.27 ml/min/1.73 m(2). 74.3% of the patients had a GFR >60 ml/min/1.73 m(2), 24% of the patients had a GFR between 30 and 60 ml/min/1.73 m(2), and 1.7% of the patients had a GFR <30 ml/min/1.73 m(2). There were significant inverse correlations between adiponectin (r=-0.372; p<0.001), resistin (r=-0.227; p<0.001) and leptin (r=-0.151; p=0.009) concentrations and GFR. After multivariate adjustment, the associations remained significant for adiponectin and resistin. Subgroup analysis in patients with GFR >60 ml/min/1.73 m(2) showed a significant correlation between GFR and adiponectin as well as leptin concentrations. However, after adjustment, these associations no longer were significant. CONCLUSIONS: There is an independent association between GFR and the serum concentrations of adiponectin and resistin. However, this association is not present at GFR >60 ml/min/1.73 m(2). This finding suggests that adipokine concentrations in mildly impaired and normal renal function are influenced by factors other than GFR.

Evaluation of two fully automated novel enzyme-linked immunosorbent assays for the determination of human adiponectin in serum

BACKGROUND: In contrast to RIA, recently available ELISAs provide the potential for fully automated analysis of adiponectin. To date, studies reporting on the diagnostic characteristics of ELISAs and investigating on the relationship between ELISA- and RIA-based methods are rare. METHODS: Thus, we established and evaluated a fully automated platform (BEP 2000; Dade-Behring, Switzerland) for determination of adiponectin levels in serum by two different ELISA methods (competitive human adiponectin ELISA; high sensitivity human adiponectin sandwich ELISA; both Biovendor, Czech Republic). Further, as a reference method, we also employed a human adiponectin RIA (Linco Research, USA). Samples from 150 patients routinely presenting to our cardiology unit were tested. RESULTS: ELISA measurements could be accomplished in less than 3 h, measurement of RIA had a duration of 24 h. The ELISAs were evaluated for precision, analytical sensitivity and specificity, linearity on dilution and spiking recovery. In the investigated patients, type 2 diabetes, higher age and male gender were significantly associated with lower serum adiponectin concentrations. Correlations between the ELISA methods and the RIA were strong (competitive ELISA, r=0.82; sandwich ELISA, r=0.92; both p<0.001). However, Deming regression and Bland-Altman analysis indicated lack of agreement of the 3 methods preventing direct comparison of results. The equations of the regression lines are: Competitive ELISA=1.48 x RIA-0.88; High sensitivity sandwich ELISA=0.77 x RIA+1.01. CONCLUSIONS: Fully automated measurement of adiponectin by ELISA is feasible and substantially more rapid than RIA. The investigated ELISA test systems seem to exhibit analytical characteristics allowing for clinical application. In addition, there is a strong correlation between the ELISA methods and RIA. These findings might promote a more widespread use of adiponectin measurements in clinical research.