Alginate microparticles as carriers for the UV filter 2-ethylhexyl 4-methoxycinnamate: Influence on photostability.

OBJECTIVE: The organic ultraviolet UVB filter 2-ethylhexyl 4-methoxycinnamate (EHMC) was encapsulated in microparticles (MPs) of sodium alginate and co-loaded with vitamin E (Vit.E) by an extrusion process using an aerodynamically assisted jetting (AAJ) methodology. The aim was to assess the effect of encapsulation concerning UVB filter release from the MPs and its photochemical stability. METHODS: The EHMC photostability was analysed by exposing the samples (both MPs in aqueous dispersion and incorporated in a cream preparation) during 1 h to simulated solar light. For the MPs (empty-MP, EHMC-MP and EHMC + Vit.E-MP), the morphology and size were characterized; while in the case of the encapsulated samples, the amount of EHMC-loading was determined. The release of EHMC was evaluated by adding EHMC-MP or EHMC + Vit.E-MP to 65% ethanol in water under mechanical stirring at 32°C. RESULTS: All MPs showed a homogeneous size distribution with a median of 90.5 ± 2.5 µm for EHMC-MP and 70.4 ± 1.14 µm for EHMC + Vit.E-MP. The encapsulation efficiency was 92.9% and 99.4% for EHMC-MP and EHMC + Vit.E-MP, respectively. The observed release from the MPs was lower than the dissolution of the pure UV filter. EHMC-MP and EHMC + Vit.E-MP were successfully incorporated into a cream formulation, with no evidence of phase separation or colour modification. Upon simulated light exposure, the photoisomerization/phototransformation of EHMC encapsulated in MPs and Vit.E-MP decreased as compared to free EHMC, both in aqueous dispersion and as a cream. The conformational ratio of the isomers (Z-/E-EHMC) was found to be the lowest in the presence of Vit.E. CONCLUSION: This work demonstrates that use of these alginate microparticulate carriers could enhance the effectiveness of sunscreen preparations containing this UVB filter.

OBJECTIF: Le filtre ultraviolet organique D'UVB 2-éthylhexyl 4-methoxycinnamate (EHMC) a été encapsulé dans des microparticules (MPs) d'alginate de sodium et co-chargé avec la vitamine E (Vit.E) par un processus d'extrusion utilisant une méthode de jet assisté aérodynamique (AAJ). L'objectif était d'évaluer l'effet de l'encapsulation concernant la libération du filtre UVB par les MPs et sa stabilité photochimique. MÉTHODES: La photostabilité EHMC a été analysée en exposant les échantillons (les deux MPs en dispersion aqueuse et incorporés dans une préparation de type crème) pendant 1 h à la lumière solaire simulée. Pour les MPs (MP-vide, EHMC-MP et EHMC - Vit.E-MP), la morphologie et la taille ont été caractérisées; tandis que dans le cas des échantillons encapsulés, la quantité de charge EHMC a été déterminée. Le relargage de l'EHMC a été évalué en ajoutant EHMC-MP ou EHMC - Vit.E-MP à 65% d'éthanol dans l'eau sous agitation mécanique à 32°C. RÉSULTATS: Tous les MP ont montré une distribution homogène de taille avec une médiane de 90,5 +- 2,5 µm pour EHMC-MP et de 70,4 +- 1,14 µm pour l'EHMC et Vit.E-MP. L'efficacité de l'encapsulation était de 92,9 % et 99,4 % pour EHMC-MP et EHMC - Vit.E-MP, respectivement. Le relargage observé des MPs était inférieur à la dissolution du filtre UV pur. EHMC-MP et EHMC - Vit.E-MP ont été incorporés avec succès dans une formulation de crème, sans aucune preuve de séparation de phase ou de modification des couleurs. Après exposition à la lumière simulée, la photoiomérisation/phototransformation de l'EHMC encapsulée dans les MPs et Vit.E-MP a diminué par rapport à l'EHMC libre, à la fois dans la dispersion aqueuse et la crème. Le rapport conformationnel des isomères (Z-/E-EHMC) s'est avéré le plus bas en présence de Vit.E.

Polyhydroxybutyrate and polyhydroxydodecanoate produced by Burkholderia contaminans IPT553.

AIMS: In this paper, we introduce a new Burkholderia contaminans capable of producing a newly characterized polymer. METHODS AND RESULTS: CG-MS and magnetic nuclear resonance 1 H and 13 C were used to determine the constitution of polymers produced in glucose, glucose with casein, sucrose and sucrose with casein. Three pairs of primers were used to find the polyhydroxyalkanoates (PHA) synthase class and sequence. The synthesized polymers were composed by short-chain length PHA (scl-PHA), especially polyhydroxybutyrate (PHB), and medium chain length PHA (mcl-PHA), especially polyhydroxydodecanoate (PHDd), and their concentration, constitution and molecular weight depend on carbon source used. The bacterium showed only class I synthase which could not explain the mcl-PHA production. CONCLUSIONS: Burkholderia contaminans has a class I PHA synthase and produces PHB combined to PHDd when cultivated in sucrose or glucose, and PHDd concentration is affected when casein is used. SIGNIFICANCE AND IMPACT OF THE STUDY: PHA are natural polymers produced by a wide range of bacteria. The presence of PHDd monomers confers to the polymer elastomeric properties. Previously, PHDd was only obtained when bacteria were cultivated in related carbon source. In this work, B. contaminansIPT553 produced PHB with PHDd using simple and low-cost carbon sources that can make possible the cheaper production of a more flexible biopolymer with crystallinity and elasticity different from the more common PHAs.

Relationship Between Pathological Findings and Cholinesterase Activity and Nitric Oxide Levels in Cattle Infected Naturally by Eurytrema coelomaticum.

The aim of this study was to evaluate the role of butyrylcholinesterase (BChE) (in the serum and pancreas), acetylcholinesterase (AChE) (in the whole blood and pancreas) and nitric oxide (NO) (in the serum and pancreas) in cattle infected naturally by Eurytrema coelomaticum. Fifty-one cattle were studied, including 33 infected by E. coelomaticum and 18 uninfected animals. Significantly greater AChE activity was found in the pancreas of infected animals (P <0.01); however, these cattle had lower AChE activity in whole blood. BChE activity was greater in the sera of infected animals (P = 0.05), but was less in pancreatic samples. NO levels were significantly higher in the sera (P <0.05) and pancreas (P <0.001) of infected cattle compared with uninfected animals. A positive correlation was found between AChE activity in the pancreas and parasite load, but there was negative correlation between pancreatic BChE activity and parasitic load. Expression of AChE, BChE and NO is therefore linked to the inflammation caused by E. coelomaticum in cattle.

Facilitation of 5-HT(2A/2C)-mediated neurotransmission in the ventromedial hypothalamic nucleus decreases anxiety in the elevated T-maze.

Previous evidence has shown that facilitation of GABA/benzodiazepine-mediated neurotransmission in the ventromedial hypothalamus (VMH) inhibits both escape and inhibitory avoidance responses generated in the elevated T-maze test of anxiety (ETM). These defensive behaviors have been associated with panic and generalized anxiety, respectively. Aside from GABA/benzodiazepine receptors, the VMH also contains a significant number of serotonin (5-HT) receptors, including 1A, 2A and 2C subtypes. The purpose of the present study was to investigate the effect of the activation of 5-HT(1A) and 5-HT(2A/2C) receptors in the VMH on defensive behavioral responses in rats submitted to the ETM. For that, male Wistar rats were treated intra-VMH with the 5-HT(1A) agonist 8-OH-DPAT, with the 5-HT(2A/2C) agonist DOI, with the 5-HT(2C) selective agonist MK-212, or with the 5-HT(2A/2C) antagonist ketanserin and 10 min after were submitted to the ETM. Results showed that both DOI and MK-212 significantly decreased avoidance measurements, an anxiolytic-like effect, without altering escape. 8-OH-DPAT and ketanserin were without effect, although the last drug attenuated the effects of DOI. None of the drugs altered locomotor activity in an open field. These results suggest that 5-HT(2A/2C) receptors of the VMH are involved in the regulation of inhibitory avoidance and might be of relevance to the physiopathology of generalized anxiety.

Diagnosis of canine leishmaniasis in the endemic area of Belo Horizonte, Minas Gerais, Brazil by parasite, antibody and DNA detection assays.

Canine leishmaniasis caused by Leishmania chagasi (L. infantum) is found throughout the South American continent, including Brazil, and dogs are considered to be the main reservoir host for this parasite. To support the implementation of a diagnostic protocol for surveillance of the disease in the region of Belo Horizonte (Minas Gerais, Brazil) we have compared the sensitivity and specificity of two serological tests, indirect immunofluorescent antibody test (IFAT) and direct agglutination test (DAT), with the combination of direct microscopy-culture-PCR as the gold standard, using samples obtained from 103 dogs in the city of Belo Horizonte, Minas Gerais. The currently used standard serodiagnostic test, IFAT, had a sensitivity of 100% and its specificity was 74% compared to the gold standard of the study. The sensitivity and specificity of the DAT were 100% and 91%, respectively. On the basis of this study it is recommended to change from the IFAT to DAT for the serodiagnosis of canine leishmaniasis because of the superior specificity of the test combined with its user-friendliness.

Development of a fast agglutination screening test (FAST) for the detection of anti-Leishmania antibodies in dogs.

A fast agglutination screening test (FAST) for the detection of anti-Leishmania antibodies in serum samples from dogs with visceral leishmaniosis was developed. The test is based on the direct agglutination test (DAT), but combines a higher parasite concentration with a smaller test volume. In contrast to the DAT, the FAST makes use of only one serum dilution and the results can be read within 3 h as opposed to 18-20 h for the DAT. The FAST was evaluated using serum samples of confirmed cases of the disease and healthy controls collected in the most important endemic regions of canine visceral leishmaniosis, import cases of canine leishmaniosis in a non-endemic country, from non-endemic healthy controls and from dogs with other diseases. The performance of the FAST was compared with standard DAT. In the present study, the FAST had a sensitivity of 93.6% and a specificity of 89.0%. The DAT had a sensitivity of 88.6% and a specificity of 96.7%. Furthermore, using a large panel of serum samples of previously examined DAT positive or negative dogs it was shown that degree of agreement between the two tests was high (95.7%; kappa value = 0.91). The FAST offers the advantages of the DAT based on freeze-dried antigen with respect to stability of the antigen, sensitivity and specificity. Moreover, the FAST allows the rapid screening of a large number of samples, which makes the test very useful for epidemiological screening of large populations of dogs.

Epidemiological studies of an outbreak of cutaneous leishmaniasis in the Rio Jequitinhonha Valley, Minas Gerais, Brazil.

We detected an outbreak of American cutaneous leishmaniasis in the Jequitinhonha River Valley, Minas Gerais, Brazil. Clinical and epidemiological aspects were studied for a period of two years. Data include results of physical examinations, Montenegro skin test and serology. In total 72 of the 299 individuals evaluated presented active lesions. Only one case out of these 72 patients showed the mucosal form of the disease. The precarious sanitary conditions, low educational level and low income found in the population studied demonstrated that, as with the other parasitic diseases, cutaneous leishmaniasis occurs with greater frequency in needy populations. A canine serological survey detected 20.3% (30/148) of dogs reactive to the Leishmania antigen. Lutzomyia intermedia was the predominant phlebotomine species and the majority of the specimens (84.9%) were captured in the peridomicile. Four samples from human and three from canine cases were isolated and characterised by PCR and isoenzymes as being Leishmania (Viannia) braziliensis. The peridomiciliary nature of the disease is discussed.

Primary aortoenteric fistula related to septic aortitis.

CONTEXT: Primary aortoenteric fistulas usually result from erosion of the bowel wall due to an associated abdominal aortic aneurysm. A few patients have been described with other etiologies such as pseudoaneurysm originating from septic aortitis caused by Salmonella. OBJECTIVE: To present a rare clinical case of pseudoaneurysm caused by septic aortitis that evolved into an aortoenteric fistula. CASE REPORT: A 65-year-old woman was admitted with Salmonella bacteremia that evolved to septic aortitis. An aortic pseudoaneurysm secondary to the aortitis had eroded the transition between duodenum and jejunum, and an aortoenteric fistula was formed. In the operating room, the affected aorta and intestinal area were excised and an intestine-to-intestine anastomosis was performed. The aorta was sutured and an axillofemoral bypass was carried out. In the intensive care unit, the patient had a cardiac arrest that evolved to death.

Aortoesophageal fistula caused by aneurysm of the thoracic aorta: successful surgical treatment, case report, and literature review.

Aortoesophageal fistula induced by atherosclerotic thoracic aortic aneurysm is rare, but is usually a fatal disorder, with few survivors reported. We report the case of a 72-year-old man with aortoesophageal fistula successfully treated in a two-stage operation. In the first stage, we performed resection and replacement of the aortic aneurysm with a prosthetic graft in situ, esophagectomy, cervical esophagostomy, and jejunostomy. After the patient recovered well postoperatively, a transmediastinal retrosternal interposition of the stomach was performed, with esophagogastroanastomosis in the cervical area, to re-establish the gastrointestinal tract. We include a discussion of the causes, diagnostic approach, management of the aorta and esophagus, and review of the literature.

Variation of infrarenal aortic diameter: A necropsy study.

PURPOSE: To determine anatomicomorphological changes in the infrarenal portion of the abdominal aorta, we performed 645 dissections of the segment in corpses undergoing necropsy. METHODS: The aortas were removed from the corpses with a surgical technique; by means of a device that we designed, the external diameter of the artery was measured after luminal pressure was reestablished. This way, it was possible to avoid underestimation of the arterial diameter postmortem. The influence of age, sex, body size, arterial hypertension, chronic obstructive pulmonary disease, and coronary disease on the aortic diameter and the influence of different degrees of sclerosis on the infrarenal aorta wall were analyzed. Considering the diameters, aortas were regarded as "normal" when they did not present any ectasia, arteriomegaly, aneurysm, or hypoplasia. RESULTS: The sample involved 645 subjects whose ages ranged from 19 to 97 years (mean age, 55.8 years). Of the 645 subjects, 65.5% (423) were men, 34.5% (222) were women, 81% (523) were white, and 19% (122) were of another race. The diameters of arteries showing no anomalous dilatation (ectasis, arteriomegaly, or aneurysm) varied according to subject age, sex, body length, and the degree of atherosclerosis on the aorta wall (P <.01). Aortic diameters of those subjects with arterial hypertension, coronary disease, and chronic obstructive pulmonary disease were compared with the aortic diameters of control subjects, and significant differences were not shown (P >.05). Twenty-nine aneurysms were found (4.5% prevalence). Four were ruptured aneurysms, and all occurred in aortas with diameters larger than 5.0 cm. CONCLUSION: The infrarenal aortic diameter enlarges with aging, and this enlargement occurs earlier in men than in women. Those subjects who had a longer body length and advanced sclerosis on the aorta wall had larger aortic diameters. There was a high prevalence of infrarenal aneurysms (4.5%), with rupture found solely in aortas with diameters larger than 5.0 cm.

Haematopoietic response and bcl-2 expression in patients with acute myeloid leukaemia.

Bcl-2 expression, the number of apoptotic cells and the growth and differentiation of early bone marrow progenitor cells were studied in patients with confirmed diagnosis of acute myeloid leukaemia (AML). Bone marrow cells from normal individuals were used as controls. We observed an increased percentage of bcl-2-mononuclear bone marrow cells expression in AML patients in relation to controls (p =0.002). Accordingly, the number of apoptotic cells was reduced (p = 0.001) and there was a negative correlation between bcl-2 expression and the number of apoptotic cells (r=-0.664, p<0.001). In addition, bcl-2 expression was significantly increased in the chemotherapy resistant group in relation to the responsive group (p = 0.03). Lower rate of survival was observed in the group of AML patients with autonomous proliferation (p=0.01). These results suggest that a high bcl-2 expression and the presence of autonomous proliferation are related to a poor prognosis in AML.

Ca2+-stimulated mitochondrial reactive oxygen species generation and permeability transition are inhibited by dibucaine or Mg2+.

Mitochondrial swelling and membrane protein thiol oxidation associated with mitochondrial permeability transition induced by Ca2+ and t-butyl hydroperoxide or inorganic phosphate, but not 4, 4'-diisothiocyanatostilbene-2,2'-disulfonic acid or phenylarsine oxide, are inhibited by the local anesthetic dibucaine. Dibucaine promotes an inhibition of the Ca2+-induced increase in mitochondrial H2O2 generation measured by the oxidation of scopoletin in the presence of horseradish peroxidase. This decrease in mitochondrial H2O2 generation may be attributed to the reduction of Ca2+ binding to the membrane induced by dibucaine, as assessed by measuring 45Ca2+ binding to the mitochondrial membrane. Mg2+ also inhibited Ca2+ binding to the mitochondrial membrane, mitochondrial swelling, membrane protein thiol oxidation, and H2O2 generation induced by Ca2+. Together, these results demonstrate that the mechanism by which dibucaine and Mg2+ inhibit mitochondrial permeability transition is related to the decrease in reactive oxygen species generation induced by Ca2+-promoted alterations of inner mitochondrial membrane properties.

A field method for the determination of whole blood cholinesterase.

Blood cholinesterase activity is an efficient indicator of exposure to organophosphate insecticides. Field methods, in spite of lacking sensitivity, are important when practical determinations and immediate results are necessary One of the mostly used field methods to assess blood cholinesterase activity is the Lovibond Cholinesterase Field Kit. This paper proposes to substitute the comparator disk of the Lovibond Field Kit with a set of standard solutions that exhibit similar colours. Dilutions of Bromothymol blue, whole blood and acetic acid in different concentrations were used to construct a set of coloured solutions which correspond to different degrees of ChE inhibition. The comparison of acetylcholinesterase activity measured with the two methods showed good agreement and satisfactory reproducibility of results. The use of a standard colored solution kit seems more suitable and manageable for field studies than the Lovibond comparator disk.