Stable thrombus formation on irradiated microvascular endothelial cells under pulsatile flow: Pre-testing annexin V-thrombin conjugate for treatment of brain arteriovenous malformations.

BACKGROUND: Our goal is to develop a vascular targeting treatment for brain arteriovenous malformations (AVMs). Externalized phosphatidylserine has been established as a potential biomarker on the endothelium of irradiated AVM blood vessels. We hypothesize that phosphatidylserine could be selectively targeted after AVM radiosurgery with a ligand-directed vascular targeting agent to achieve localized thrombosis and rapid occlusion of pathological AVM vessels. OBJECTIVE: The study aim was to establish an in vitro parallel-plate flow chamber to test the efficacy of a pro-thrombotic conjugate targeting phosphatidylserine. METHODS: Conjugate was prepared by Lys-Lys cross-linking of thrombin with the phosphatidylserine-targeting ligand, annexin V. Cerebral microvascular endothelial cells were irradiated (5, 15, and 25 Gy) and after 1 or 3 days assembled in a parallel-plate flow chamber containing whole human blood and conjugate (1.25 or 2.5 µg/mL). Confocal microscopy was used to assess thrombus formation after flow via binding and aggregation of fluorescently-labelled platelets and fibrinogen. RESULTS AND CONCLUSIONS: The annexin V-thrombin conjugate induced rapid thrombosis (fibrin deposition) on irradiated endothelial cells under shear stress in the parallel-plate flow device. Unconjugated, non-targeting thrombin did not induce fibrin deposition. A synergistic interaction between radiation and conjugate dose was observed. Thrombosis was greatest at the highest combined doses of radiation (25 Gy) and conjugate (2.5 µg/mL). The parallel-plate flow system provides a rapid method to pre-test pro-thrombotic vascular targeting agents. These findings validate the translation of the annexin V-thrombin conjugate to pre-clinical studies.

Increased immunoglobulin production in silver catfish (Rhamdia quelen) exposed to agrichemicals.

Fish vaccination has been increasingly exploited as a tool to control pathogen infection. The production of immunoglobulin following vaccination might be affected by several factors such as management procedures, water temperature, and the presence of xenobiotics. In the present study, we aimed to investigate the kinetics of immunoglobulin production in silver catfish (Rhamdia quelen) inoculated with inactivated Aeromonas hydrophila and kept at two different water temperatures (17.4±0.46 or 21.3±0.36C). The effect of a second antigen inoculation and exposure of fish to sublethal concentrations of the herbicides atrazine and glyphosate at 10% of the lethal concentration (LC50-96h) on specific serum antibodies were also investigated. Antibodies to A. hydrophila were detected as early as 7 days post-inoculation and increased steadily up to 35 days. The kinetics of antibody production were similar in fish kept at 17.4±0.46 and 21.3±0.36C, and reinoculation of antigen at 21 days after priming failed to increase specific antibody levels. Intriguingly, we found that, in fish exposed to atrazine and glyphosate, the secretion of specific antibodies was higher than in non-exposed inoculated fish. These findings are important for the design of vaccines and vaccination strategies in Neotropical fish species. However, because atrazine and glyphosate are widespread contaminants of soil and water, their immune-stimulating effect could be harmful, in that fish living in herbicide-contaminated water might have increased concentrations of nonspecific antibodies that could mediate tissue injury.

Hepatitis C: a critical analysis of therapeutic response predictors.

Treatment of chronic hepatitis C is still unspecific. However, there is great expectancy concerning the new pegylated interferons. As there has been much controversy about the best parameters to determine whether treatment is effective, we analyzed several criteria currently used for evaluation, including serum alanine aminotransferase (ALT) normalization, viral load reduction and improvement of hepatic histology.

Hepatitis C: a critical analysis of therapeutic response predictors

Treatment of chronic hepatitis C is still unspecific. However, there is great expectancy concerning the new pegylated interferons. As there has been much controversy about the best parameters to determine whether treatment is effective, we analyzed several criteria currently used for evaluation, including serum alanine aminotransferase (ALT) normalization, viral load reduction and improvement of hepatic histology.

[Mercury levels in fish consumed by the Sai Cinza indigenous community, Munduruku Reservation, Jacareacanga County, State of Pará, Brazil]. / Níveis de mercúrio em peixes consumidos pela comunidade indígena de Sai Cinza na Reserva Munduruku, Município de Jacareacanga, Estado do Pará, Brasil.

This study evaluated fish consumption and mercury levels in fish consumed by an indigenous community in the State of Pará. Eighty fish samples were collected (barbado, surubim, traíra, tucunaré, piranha, aruanã, caratinga, aracu, mandiá, jandiá, and pacu). Mercury analysis was performed using a Mercury Analyzer HG-3500. Average mercury concentration in carnivorous species was 0.293 (g/g (SD=0.104), while in non-carnivorous species it was 0.112 (g/g (SD=0. 036). Brazilian legislation establishes a maximum permissible limit of 0.5 (g/g for fish consumption. No significant correlation was found between fish length or weight and mercury concentration. Types of fish most frequently consumed by the community were tucunaré, pacu, jaraqui, traíra, aracu, matrinchã, and caratinga. Carnivorous species, especially tucunaré and traíra, amongst the most frequently eaten, had higher mercury levels than non-carnivorous species. Frequency of consumption is crucial to assess the risk of mercury contamination in communities who lack alternative food sources.