Photochemical inactivation of viruses and bacteria in platelet concentrates by use of a novel psoralen and long-wavelength ultraviolet light

BACKGROUND: A photochemical treatment process has been developed for the inactivation of viruses and bacteria in platelet concentrates. This process is based on the photochemical reaction of a novel psoralen, S- 59, with nucleic acids upon illumination with long-wavelength ultraviolet light (UVA, 320–400 nm). STUDY DESIGN AND METHODS: High levels of pathogens were added to single-donor platelet concentrates containing 3 to 5 × 10(11) platelets in 300 mL of 35-percent autologous plasma and 65-percent platelet additive solution. After treatment with S-59 (150 microM) and UVA (0-3 J/cm2), the infectivity of each pathogen was measured with established biologic assays. In vitro platelet function after photochemical treatment was evaluated during 7 days of storage by using a panel of 14 assays. The in vivo recovery and life span of photochemically treated platelets were evaluated after 24 hours of storage in a primate transfusion model. RESULTS: The following levels of pathogen inactivation were achieved:>10(6.7) plaque-forming units (PFU) per mL of cell-free human immunodeficiency virus (HIV),>10(6.6) PFU per mL of cell-associated HIV,>10(6.8) infectious dose (ID50) per mL of duck hepatitis B virus (a model for hepatitis B virus),>10(6.5) PFU per mL of bovine viral diarrhea virus (a model for hepatitis C virus),>10(6.6) colony-forming units of Staphylococcus epidermidis, and>10(5.6) colony-forming units of Klebsiella pneumoniae. Expression of integrated HIV was inhibited by 0.1 microM S- 59 and 1 J per cm2 of UVA. In vitro and in vivo platelet function were adequately maintained after antiviral and antibacterial treatment. CONCLUSION: Photochemical treatment of platelet concentrates offers the potential for reducing transfusion-related viral and bacterial diseases.     

Hypertensive crises in patients with arterial hypertension in ambulatory treatment

This study assessed the sociodemographic characteristics and the characteristics of therapeutic adhesion of 27 bearers of arterial hypertension undergoing ambulatorial treatment who had hypertensive urgencies crises or emergencies in the city of Fortaleza in the period between October of 2002 and May of 2003. The majority were women, between 50 and 60-years old, with little formal education, treatment time shorter than 5 years and time of diagnosis varying from 5 to 10 years. The use of medicine was the treatment that was most mentioned, followed by the reduction of the consumption of salt and attendance to medical appointments. However, attending the appointments and receiving orientation did not seem to change their behavior, since most of the patients that were interviewed practiced no physical exercises and demonstrated little knowledge of the illness, for they attributed the rise of the arterial pressure to emotional factors.     

Mechanisms of increased survival after lipopolysaccharide-induced endotoxic shock in mice consuming olive oil-enriched diet

We examined the impact of dietary fatty acid intake on lipopolysaccharide (LPS)-induced endotoxic shock. C57Bl/6J mice were fed for 6 weeks with a commercial laboratory chow (CC) or with test chows containing 7% (w/w) canola oil (CO), sesame oil (SeO), soybean oil (SO), or virgin olive oil (OO). The increase in body weight and energy consumption were similar for all diets tested. In the sixth week, mice were injected intraperitoneally with 400 microg of bacterial LPS to induce endotoxic shock. LPS induced a massive neutrophil infiltration into the peritoneal cavity and an increase in lipid body (LB) formation in leukocytes recovered from the peritoneal fluid of mice fed with CC, CO, SeO, or SO. In addition, there were increases in prostaglandin E(2) (PGE(2)), leukotriene B4 (LTB(4)), and cytokines IL-6, IL-10, and MCP-1 in peritoneal lavage, as well as in plasma TNF-alpha. In contrast, mice fed with OO exhibited reduced neutrophil accumulation and LB formation, and also had lower levels of PGE(2), LTB(4), MCP-1, and TNF-alpha. All mice fed with CC, CO, SeO, or SO died within 48 to 72 h after LPS injection. Interestingly, mice fed with the OO diet were resistant to endotoxic shock, with 60% survival at 168 h. These data indicate that intake of OO may have a beneficial role, reducing the magnitude of the inflammatory process triggered by endotoxic shock through modulation of LB formation and of the production of inflammatory mediators.     

Fluctuating Asymmetry in Insecticide-Resistant and Insecticide-Susceptible Strains of the Maize Weevil, <Emphasis Type="Italic">Sitophilus zeamais</Emphasis> (Coleoptera: Curculionidae)

Insecticide resistance is an evolutionary response of specific insect populations subjected to exposure and consequent selection by an insecticide. As such, this phenomenon is important as a biomonitoring strategy and also has economical importance in the case of insect-pests by compromising their control. Fluctuating asymmetry (FA) of bilateral traits is a measure of developmental instability also suggested as a monitoring tool for environmental pollution with potential consequences for fitness. Responses to selective agents might have pleiotropic effects influencing development and phenotype, which has yet to be examined for agricultural insecticides and insect-pests. Higher levels of FA are expected in the insecticide-resistant strains, which are usually at a selective disadvantage, relative to susceptible strains, in the absence of the insecticide. Two insecticide-resistant strains and an insecticide-susceptible strain of the maize pest insect Sitophilus zeamais (Coleoptera: Curculionidae) were subjected to FA measurements in 12 traits of wing veins and tibias of 100 individuals of both sexes of each strain. The insecticide-resistant strains showed lower FA than the susceptible strain, in contrast with the initial expectation. An extended period of insecticide selection probably led to the evolution of fitness-modifier genes improving the performance of the resistant genotypes, reducing their FA levels, and leading to their eventual fixation in the population. In addition, one insecticide-resistant strain and the insecticide-susceptible strain showed significant differences in FA between sexes, with more symmetrical males suggesting possible sexual selection by the females. The observed results have potential consequences for insecticide-resistance evolution and dispersal.     

Chemical characterization and in vitro cyto- and genotoxicity of ‘legal high’ products containing Kratom (<Emphasis Type="Italic">Mitragyna speciosa</Emphasis>)

Kratom is a popular ‘legal high’ mainly constituted by alkaloids extracted from the Mitragyna speciosa plant with mitragynine (MG) as the dominant active substance. The increasing use of Kratom for recreational purposes has alerted risk assessment bodies of the lack of information on the real composition and its potential health risks. The present study aimed to determine and compare the MG composition of 13 commercial products of Kratom sold online and in “smartshops”, by gas chromatography–mass spectrometry. For the first time, the cytotoxicity induced by pure MG and Kratom, extracts was evaluated in in vitro models of human intestinal (Caco-2) and neuronal (SH-SY5Y) cells after 6 and 24 h. Genotoxicity was also evaluated in intestinal Caco-2 cells following 24 h of exposure to subtoxic concentrations using the comet assay. The obtained results revealed an inconsistency between the information (‘power’) provided in labels and the MG content. Cytotoxicity tests revealed a concentration-dependent decrease in cell viability in both cellular models, with the SH-SY5Y cells being more sensitive to the Kratom extracts. The resin and the ‘powered extracts’ were the most cytotoxic samples, with IC₅₀ values significantly lower than the leaf extracts and pure MG (P < 0.0001 vs. leaf extracts and MG). In addition, significant DNA damage was observed in Caco-2 cells exposed to these extracts but not to pure MG, which suggests that other substances present in the extracts or interactions involving Kratom components might be responsible for the observed effects.     

Chiral enantioresolution of cathinone derivatives present in “legal highs”, and enantioselectivity evaluation on cytotoxicity of 3,4-methylenedioxypyrovalerone (MDPV)

Recently, great interest has been focused on synthetic cathinones since their consumption has increased exponentially. All synthetic cathinones exist as chiral molecules; the biological and/or toxicological properties of cathinones generally differ according to the enantiomers in human body. In this study, a chiral liquid chromatography method was developed to separate and determine the enantiomeric ratio of synthetic cathinones present in “legal highs” acquired in old smart shops or over the Internet. All the synthetic cathinones were efficiently enantio-separated with α and Rs ranging from 1.24 to 3.62 and from 1.24 to 10.52, respectively, using polysaccharide-based chiral stationary phases. All synthetic cathinones, with the exception of 4-methylethcathinone (4-MEC), were present in the commercialized “legal highs” in an enantiomeric proportion of 50:50. One of the studied chiral compounds was 3,4-methylenedioxypyrovalerone (MDPV), one of the most consumed cathinone derivative worldwide. Our research group has recently reported its hepatotoxicity in the racemic form. Thus, the analytical enantioresolution of the MDPV was scaled up to multi-milligram using a semi-preparative amylose tris-3,5-dimethylphenylcarbamate column (20 cm × 7.0 mm ID, 7 µm particle size). Both enantiomers were isolated with high enantiomeric purity (enantiomeric excess > 99 %). The toxicity of S-(?)-MDPV and R-(+)-MDPV was evaluated, for the first time, using primary cultures of rat hepatocytes. It was also possible to verify that MDPV enantiomers showed hepatotoxicity in a concentration-dependent manner, but displayed no enantioselective toxicity in this cell culture model.     

Zoonotic vaccinia virus infection in Brazil: clinical description and implications for health professionals

Bovine vaccinia virus outbreaks have been occurring in different regions of Brazil. We report here the time course of natural human infection by vaccinia virus and describe important clinical and epidemiological aspects of this zoonotic infection. The diagnosis of vaccinia virus infection was based on clinical, serological, and molecular procedures.