Inclusion complex and nanoclusters of cyclodextrin to increase the solubility and efficacy of albendazole.

Albendazole (ABZ), a benzimidazole widely used to control gastrointestinal parasites, is poorly soluble in water, resulting in variable and incomplete bioavailability. This has favored the appearance ABZ-resistant nematodes and, consequently, an increase in its clinical ineffectiveness. Among the pharmaceutical techniques developed to increase drug efficacy, cyclodextrins (CDs) and other polymers have been extensively used with water-insoluble pharmaceutical drugs to increase their solubility and availability. Our objective was to prepare ABZ formulations, including ß-cyclodextrin (ßCD) or hydroxypropyl-ß-cyclodextrin (HPßCD), associated or not to the water-soluble polymer polyvinylpyrrolidone (PVP). These formulations had their solubility and anthelmintic effect both evaluated in vitro. Also, their anthelmintic efficacy was evaluated in lambs naturally infected with gastrointestinal nematodes (GIN) through the fecal egg count (FEC) reduction test. In vitro, the complex ABZ/HPßCD had higher solubility than ABZ/ßCD. The addition of PVP to the complexes increased solubility and dissolution rates more effectively for ABZ/HPßCD than for ABZ/ßCD. In vivo, 48 lambs naturally infected with GIN were divided into six experimental groups: control, ABZ, ABZ/ßCD, ABZ/ßCD-PVP, ABZ/HPßCD, and ABZ/HPßCD-PVP. Each treated animal received 10 mg/kg of body weight (based on the ABZ dose) for three consecutive days. After 10 days of the last administered dose, treatment efficacy was calculated. The efficacy values were as follows: ABZ (70.33%), ABZ/ßCD (85.33%), ABZ/ßCD-PVP (82.86%), ABZ/HPßCD (78.37%), and ABZ/HPßCD-PVP (43.79%). In vitro, ABZ/HPßCD and ABZ/HPßCD-PVP had high solubility and dissolution rates. In vivo, although the efficacies of ABZ/ßCD, ABZ/ßCD-PVP, and ABZ/HPßCD increased slightly when compared to pure ABZ, this increase was not significant (P > 0.05).

Desempenho do polvo Octopus vulgaris alimentado com monodieta de mexilhão (Perna perna) / Performance of the Octopus vulgaris octopus fed on mussel (Perna perna) as monodiet

The octopus Octopus cf. vulgaris is a potential species to diversify aquaculture. Due to absence of balanced commercial diet, growth of the O. cf. vulgaris is based on natural diet with local and low-cost inputs. In Brazil, studies on experimental octopus ongrowing are recent and there is little available data. We evaluated the performance, survival and food consumption of O. vulgaris fed on mussel Perna perna for 20 days. Six octopuses with initial weight of 415±12.73g (mean±standard deviation) were divided into two groups (n=3 octopuses/group) according to the diet: MC Group (frozen mussels) and MV Group (live mussels). The Weight Gain of octopuses was 273.33±94.52g and 340.00±26.46g; the Absolute Growth Rate was 13.67±4.73 and 17.00±1.32g.dia-1 and the Specific Growth Rate of 2.95±0.58 and 2.64±0.37%.day-1 to MC and MV groups, respectively. There was no significant difference in performance between groups and the survival rate was 100%. Octopuses well accepted both diets and despite the amount of frozen mussels (129±31) was higher than in live mussels (100±19), there was no significant difference regarding the consumption between groups. Our results demonstrate that the mussel Perna perna can be used frozen or live as monodiet in O. cf. vulgaris ongrowing.(AU)

Doctors' awareness concerning primary immunodeficiencies in Brazil.

BACKGROUND: PIDs are a heterogeneous group of genetic illnesses, and delay in their diagnosis is thought to be caused by a lack of awareness among physicians concerning PIDs. The latter is what we aimed to evaluate in Brazil. METHODS: Physicians working at general hospitals all over the country were asked to complete a 14-item questionnaire. One of the questions described 25 clinical situations that could be associated with PIDs and a score was created based on percentages of appropriate answers. RESULTS: A total of 4026 physicians participated in the study: 1628 paediatricians (40.4%), 1436 clinicians (35.7%), and 962 surgeons (23.9%). About 67% of the physicians had learned about PIDs in medical school or residency training, 84.6% evaluated patients who frequently took antibiotics, but only 40.3% of them participated in the immunological evaluation of these patients. Seventy-seven percent of the participating physicians were not familiar with the warning signs for PIDs. The mean score of correct answers for the 25 clinical situations was 48.08% (±16.06). Only 18.3% of the paediatricians, 7.4% of the clinicians, and 5.8% of the surgeons answered at least 2/3 of these situations appropriately. CONCLUSIONS: There is a lack of medical awareness concerning PIDs, even among paediatricians, who have been targeted with PID educational programmes in recent years in Brazil. An increase in awareness with regard to these disorders within the medical community is an important step towards improving recognition and treatment of PIDs.

Evaluation of Cymbopogon schoenanthus essential oil in lambs experimentally infected with Haemonchus contortus.

Hematophagous gastrointestinal parasites cause significant economic losses in small ruminant grazing systems. The growing reports of multi-drug resistant parasites call for intensive research on alternative treatments for anthelmintics to help small ruminants cope with these parasites. Two-month-old lambs with mean body weight (BW) of 22.5 kg were experimentally infected with a multidrug-resistant Haemonchus contortus strain. Infected animals were dosed orally with Cymbopogon schoenanthus essential oil to evaluate its anthelmintic potential. Eighteen animals were allocated into three groups of six animals, and each received one of the following treatments: Group 1 - control (10 mL of water), Group 2 - C. schoenanthus essential oil (180 mg/kg BW); and Group 3 - C. schoenanthus essential oil (360 mg/kg BW). Animals received the oil once a day for 3 consecutive days. Lambs were evaluated clinically for blood biochemistry before, at 1, 5, 10, 15 and 20 days after treatment, and then were euthanized to assess the total worm burden. No statistically significant reduction in fecal egg count, packed cell volume or total worm count was observed after treatments. Also, no statistical difference among group means for blood levels of urea, creatinine, albumin, alkaline phosphatase, aspartate aminotransferase and gamma glutamyl transferase was found. Larval development assay (LDA) and egg hatch assay (EHA) were performed from feces of treated animals at 1, 5, 10 and 15 days after essential oil administration. An inhibition in LDA was observed 1 day after the 3-day treatment in larvae from feces of animals treated with 360 mg/kg essential oil. In conclusion, the essential oil at the doses of 180 mg/kg and 360 mg/kg was safe to sheep, but failed as an anthelmintic treatment when applied to young sheep artificially infected with a multidrug-resistant H. contortus strain.

Larval settlement and spat recovery rates of the oyster Crassostrea brasiliana (Lamarck, 1819) using different systems to induce metamorphosis.

This study aimed at the assessment, in the laboratory, of the larval settlement and spat recovery rates of oysters of the species Crassostrea brasiliana using plastic collectors, epinephrine (C9H13NO3 C4H6O6) and shell powder in settlement tanks. Polypropylene was used attached to bamboo frames. The material was chosen due to its pliability--that favours the spat detachment. Two experiments were carried out; the first between February and April 2008, and the second between November and December 2008 at the Marine Mussel Laboratory of Santa Catarina Federal University (Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina). In the first experiment, the scratched plastic collectors were tested consorting them with shell powder; on the second, the plastic collectors were tested consorted with shell powder, only shell powder and epinephrine as the metamorphosis stimulator. The quantification was carried out of the larvae settled in the plastic collectors, and of the recovery and integrity of the spats after their detachment. The first experiment has shown a recovery rate of 48.83% of the spats in comparison with the D larvae used. From this percentage, 4.9% settled in the plastic collectors and 43.93% in shell powder. The second experiment revealed 55.78% regarding the settled spats in comparison with the total of larvae used (using epinephrine), 78.62% in the treatment with the collector plus shell powder and 58.33% in the treatment only with shell powder. Thus, the use of the collector plus shell powder resulted in a greater spat recovery when compared to the other treatments.

Sexual stages of the female portion in the scallop Nodipecten nodosus (Linné, 1758) and astaxanthin quantity in each stage / Estágios sexuais da porção feminina da gônada da vieira Nodipecten nodosus (Linné, 1758) e a quantidade de astaxantina em cada estágio

This work describes the gametogenic cycle of the scallop Nodipecten nodosus kept in a culture system. To this end, during one year, samples were taken from the broodstocks every 30 days to be submitted to macroscopic and microscopic analyses and to measure the amount of astaxanthin. To perform the microscopic evaluation, 5 μ slices from the median portion of the female part of the gonad were submitted to the pattern methodology for histological analyses with paraffin and HE coloration. The remaining portion of the female gonad was lyophilised to extract and quantify the levels of astaxanthin using HPLC. The microscopic analyses revealed four well defined stages for the reproductive cycle. Analyses of data taken throughout the year indicated preferential spawning periods from December to January and from July to September. The astaxanthin analyses showed higher amounts of this carotenoid during the advanced pre-spawning and the initial spawning periods than during gametogenesis, initial pre-spawning, advanced spawning, and the spent stages. According to these results, it was possible to establish a descriptive table of the sexual stages of the female portion of the gonad and the amount of astaxanthin in the sexual stage of the scallop Nodipecten nodosus.

Este trabalho descreve o ciclo gametogênico da vieira Nodipecten nodosus mantida em ambiente de cultivo. Para isto, durante um ano, amostras de indivíduos reprodutores foram coletadas a cada 30 dias e submetidas à avaliação macroscópica e microscópica e à quantificação de astaxantina. Para a avaliação microscópica, secções de 5 μ da porção mediana feminina da gônada foram submetidas à metodologia de análise histológica padrão em parafina e coloração HE. O restante da porção feminina da gônada foi liofilizado para extração e quantificação de astaxantina em HPLC. A avaliação microscópica permitiu a descrição de quatro estágios bem definidos para o ciclo reprodutivo. Na análise ao longo do ano, foram observados períodos preferenciais de desova em dezembro e janeiro e de julho a setembro. A análise da quantidade de astaxantina, mostrou, nos estádios de pré-desova avançada e de desova inicial, uma maior quantidade desse carotenoide em comparação aos estádios de gametogênese, pré-desova inicial, desova avançada e repouso. Em função desses resultados, foi possível estabelecer um quadro descritivo dos estágios sexuais da porção feminina da gônada e quantidade de astaxantina em cada estágio sexual da vieira Nodipecten nodosus.

Carotenoid extraction from the gonad of the scallop Nodipecten nodosus (Linnaeus, 1758) (Bivalvia: Pectinidae) / Extração de carotenóides da gônada da vieira Nodipecten nodosus (Bivalvia: Pectinidae)

In marine bivalve mollusks, unsaturated molecules called carotenoids are present in the natural diet and play an important role in different biological process, especially in reproduction. In order to gain more insights into these compounds in Nodipecten nodosus it was necessary to develop a suitable protocol for extraction of carotenoids from the gonads. Female gonads of cultured scallops (75 mm length) were lyophilized and macerated in liquid N2. To verify the effect of composition in organosolvents on the extracting solutions, two organic solvents were tested: acetone and hexane (Ac = O:Hex) at four ratios, 1:1, 1:3, 1:5, and 2:3, in four static extraction times: 0, 5, 10, and 15 minutes. Total carotenoids and astaxanthin contents were determined in the crude extracts by UV-visible spectrophotometry and high performance liquid chromatography (HPLC), respectively. Triplicate aliquots of 50 mg were used for each treatment. The results indicated that the best single extraction (0.312 ± 0.016 µg carotenoids/mg) was attained with Ac = O: Hex 1:3, for 15 minutes. Through exhaustive extraction methodology (10x), a superior yield (0.41 ± 0.001 µg carotenoids/mg) was obtained from a gonad sample in comparison to the highest value found for a single extraction. Astaxanthin content was reduced by 8.6 percent in carotenoid extract preservation assay, i.e., -18 °C, 26 days incubation, under N2 atmosphere.

Em moluscos bivalves marinhos, carotenóides insaturados estão presentes na dieta natural, com um importante papel em diversos processos biológicos, em especial na reprodução. A elucidação dos efeitos destes compostos em Nodipecten nodosus requer o desenvolvimento de um protocolo adequado para a extração de carotenóides das gônadas desses animais. Para isso, gônadas de vieiras cultivadas (75 mm de comprimento) foram liofilizadas e maceradas em N2 líquido. Amostras em triplicata com 50 mg foram coletadas para a utilização em cada tratamento. Os conteúdos de carotenóides totais e astaxantina foram determinados via espectrofotometria de luz UV-visível e cromatografia líquida de alta eficiência (CLAE), respectivamente. O efeito da composição em organosolventes das soluções de extração foi testado utilizando-se acetona (Ac = O) e hexano (Hex) em quatro proporções (Ac = O:Hex): 1:1, 1:3, 1:5, e 2:3; em quatro tempos de extração: 0, 5, 10, e 15 minutos. Os resultados mostraram que o melhor rendimento de extração (0,312 ± 0,016 µg carotenóides/mg) foi obtido com Ac = O:Hex, 1:3, por 15 minutos. Com a utilização de protocolo de extração exaustiva (10x), uma quantidade superior (0,41 ± 0,001 µg de carotenóides/mg) foi obtida de amostras de gônada, comparativamente aos valores obtidos em extrações únicas. O conteúdo de astaxantina foi reduzido em 8,6 por cento em testes de preservação deste metabólito em extratos crus (-18 °C, 26 dias de incubação em atmosfera de N2).

Micronucleus induction in mussels exposed to okadaic acid.

Some toxins present in the marine environment are capable of inducing mutagenicity and/or carcinogenicity. Among these toxins, okadaic acid (OA) is gaining considerable interest since it induces DNA based modifications at low concentrations and accumulates in filter-feeding marine animals, including those used for human consumption. This study aims to evaluate the genotoxicity of OA in the haemocytes of the mussel Perna perna, using the micronucleus assay. Fifty-four mussels were separated into three groups of 18 animals. One group received 0.3 microg of OA diluted in 10 microl of ethanol and ultrapure water while the other groups were considered as controls and were exposed to a solvent plus seawater mixture. A significantly higher frequency of micronuclei was observed in haemocytes from the OA-exposed group. There were no statistical differences between the two control groups.

[Comparative study between verapamil and adenosine triphosphate in the treatment of paroxysmal supraventricular tachycardia]. / Estudo comparativo entre verapamil e adenosina-trifosfato no tratamento das taquicardias paroxísticas supraventriculares.

PURPOSE: To evaluate the efficacy and safety of intravenous (IV) adenosine-triphosphate (ATP) and verapamil to convert acute episodes of paroxysmal supraventricular tachycardia (PSVT) to sinus rhythm. METHODS: Fifty patients with PSVT were randomized in two groups: A) 25 treated with IV bolus of ATP (10 or 20mg), and V) 25 treated with IV verapamil, up to 15mg, during 3min. We evaluated the time delay necessary to convert the arrhytmia, doses, and side-effects. Patients with acute ischemic syndromes (< 3 weeks), severe congestive heart failure, and treatment with dipyridamole or methylxanthine were excluded. RESULTS: There were no differences between the two groups regarding to age, sex, and success rate. The average time till reversal were respectively, 30s and 248s for ATP and verapamil. Ventricular ectopy and general discomfort were observed in 33% of patients receiving ATP, whereas no side-effects occurred in group V. CONCLUSION: ATP is a good option to convert rapidly PSVT to sinus rhythm and, probably, could be the first choice to treat PSVT patients with ventricular dysfunction.

Estudo comparativo entre Verapamil e Adenosina-Trifosfato no tratamento das taquicardias paroxísticas supraventriculares / Adenosin-three phosphate compared with Verapamil to treat paroxysmal supraventricular tachycardia

Objetivo - comparar a eficácia e segurança da adenosina-trifosfato (ATP) e verapamil endovenosos na reversäo das taquicardias paroxísticas supraventriculares (TPSV). Métodos - foram analisados 50 pacientes com diagnóstico de TPSV pelo eletrocardiograma convencional. Excluíram-se aqueles com insuficiência cardíaca, quadro isquêmico agudo, uso de dipiridamol e metilxantinas. Os pacientes foram distribuídos por sorteio em dois grupos: A) ATP em bolo de 10 ou 20 mg; V) verapamil em infusäo rápida de at15 mg. Avaliaram-se sucesso, tempo de reversäo, dose empregada e efeitos colaterais. Resultados - Ocorreu sucesso na reversão em 96% do grupo A e em 92 % do grupo V (p= 1,0). O tempo médio de reversão foi de 30s no grupo A e 248s no V. Efeitos colaterais foram encontrados em 33% no grupo A, e não ocorreram no grupo V Conclusão - O ATP é uma opção atraente para a reversão das TPSV, apresentando efeitos colaterais fugazes, podendo ser droga de escolha no tratamento desses episódios na presença de disfunção ventricul

Purpose - To evaluate the efficacy and safety of intravenous (IV) adenosine-triphosphate (ATP) and verapamil to convert acute episodes of paroxysmal supraventricular tachycardia (PSVT) to sinus rhythm. Methods - Fifty patients with PSVT were randomized in two groups: A) 25 treated with IV bolus of ATP (10 or 20mg), and V) 25 treated with IV verapamil, up to 15mg, during 3min. We evaluated the time delay necessary to convert the arrhytmia, doses, and side-effects. Patients with acute ischemic syndromes (<3 weeks), severe congestive heart failure, and treatment with dipyridamole or methylxanthine were excluded. Results - There were no differences between the two groups regarding to age, sex, and success rate. The average time till reversal were respectively, 30s and 248s for ATP and verapamil. Ventricular ectopy and general disconfort were observed in 33% of patients receiving ATP, where as no side-effects occurred in group V. Conclusion - ATP is a good option to convert rapidly PSVT to sinus rhythm and, probably, could be the first choice to treat PSVT patients with ventricular dysfunction

Iron from complex salts and its bioavailability to rats.

Complex iron compounds are being used as iron suppliers. Bioavailability of their iron was tested through several parameters. A biological iron prophylactic study in rats was simultaneously carried out with three of these iron complexes products: ferric orthophosphate, iron sodium EDTA and iron glycine chelate. Their iron biodisponibility was compared to that of ferrous sulfate. Five low iron diets were prepared according to AOAC method. Four of them were enriched to a level of 20 mg Fe/kg, with ferrous sulfate and the 3 iron complexes salts under study. These diets were offered to 5 groups of 6 weanling rats each in an iron prophylactic test. Food intake was measured during 5 weeks, weight checked weekly, blood and liver collected for analyses. Weight gain, hemoglobin, hematocrit, transferrin saturation, iron hemoglobin, biodisponibility and relative iron biological values were calculated. The prophylactic iron rat assay proved to be a feasible and practical model to test and compare iron salts biodisponibility. NaFeEDTA and iron aminochelate produced similar results as ferrous sulfate and their iron has a high biological value for the rats. Orthophosphate iron had lower biological value when compared to the reference standard ferrous sulfate and the other complex products studied.

Simultaneous visualization of neuropeptide and acetylcholinesterase nerve subpopulations in the perivascular plexus.

A simple method combining indirect immunofluorescence and histochemical techniques was developed in order to demonstrate the presence of both neuropeptide immunoreactivity and acetylcholinesterase activity in the same whole-mount preparation. It was found that the two methods can be combined without interfering with one another and may be viewed and photographed simultaneously. The guinea pig basilar artery was chosen as a model tissue. While vasoactive intestinal polypeptide immunoreactivity and acetylcholinesterase activity were found to occur in the same perivascular nerve fibres, tyrosine hydroxylase, neuropeptide tyrosine and calcitonin gene-related peptide immunoreactivity were present in distinct nerve subpopulations. It is possible using this double staining procedure, to analyse the interrelationship of putative cholinergic nerves with other components of the autonomic and sensory nervous system.

[The use of drinking water as a vehicle of nutrients: experimental studies with iron]. / Utilização da água potável como veiculo de nutrientes: estudos experimentais com ferro.

This investigation is part of a large project, the objective of which is to study the possibility of utilizing drinking water as a carrier of nutrients, such as iron. Various iron salts and in different concentrations, were added to water and tested for their effect on color and turbidity. The bioavailability of various forms of iron salts added to drinking water was tested using the rat bioassay technique. The results of this study indicated that ferric ammonium citrate changed very little the color and turbidity of the water solutions. It was considered the best suitable form of iron to be added to drinking chloride water. Ferric ammonium citrate, ferrous sulfate and ferrous gluconate were all found effective in preventing rat anemia when added to their drinking water.

Immunocytochemical study of tissue distribution and hormonal control of chondroitin-, dermatan- and keratan sulfates from rodent uterus.

The distribution patterns of rat and mouse uterine glycosaminoglycans (GAGs), as well as their modulation by estradiol (E2) and/or progesterone (P), were investigated using monoclonal antibodies (MABs) directed against chondroitin- (CS)/dermatan sulfates (DS), keratan sulfate (KS) and a trophoblast GAG. The localization of GAGs in relation to collagens (I, IV and VI) and fibronectin was also analyzed. We found that uterine GAGs are differentially distributed in the endometrium and myometrium, in a pattern that is species-related. CS-containing proteoglycans (PGs) occur between collagen bundles and fibroblasts, at the periphery of the latter, and in basement membrane zones (BMZs), in a pattern resembling that of collagen VI. BMZs contain preferentially CS-PGs bearing 4-sulfated disaccharides adjacent to the core protein. DS-PGs are mostly associated with collagen bundles. E2 and/or P elicit distinct modifications on the above described pattern, which are also species-related. The simultaneous administration of E2 and P changes the prevalent sulfation of the disaccharides adjacent to the core protein of stromal CS-PGs. In the mouse, an unsulfated intracellular epitope appears following E2 (or E2P) administration, mostly in epithelial cells. In the rat, KS and the trophoblast GAG are E2-dependent and down-regulated by P. The functional significance of the hormone-induced GAG changes, namely the possible role of the E2-dependent KS in implantation, are discussed.

Cytokeratin filaments are present in golden hamster oocytes and early embryos.

Light and electron microscope methods were used to study cytokeratin expression in the recently ovulated oocytes, fertilized eggs and early embryos from the golden hamster. Two cytokeratin polypeptides (Mr 51,000 and 58,000) were detected in oocyte lysates by immunoblotting using a polyclonal antiserum to prekeratin. In the oocyte, cytokeratin occurred as patchy aggregates consisting of short anastomosing 10-nm filaments that formed tight meshworks distributed throughout the cytoplasm. After fertilization the aggregates appeared to merge, becoming larger and concentrated at the cortical region. Prominent immunofluorescent fibrils were seen interconnecting the aggregates. In the 2-, 4- and 8-cell embryos, networks of cytokeratin filaments extended throughout the cortical and perinuclear regions, while aggregates progressively disappeared.

Filamentous cross-bridges link intermediate filaments to the nuclear pore complexes.

Intermediate filament-nuclear matrix interactions were studied in cultured rat ventral prostate cells and isolated rat uterine epithelial cells. Cytokeratin filaments were identified by immunoelectron microscopy. In addition to conventional thin section of Triton X-100 treated cells, subcellular residues composed of intermediate filaments and nuclear matrix were critical-point dried and platinum-carbon replicated. The results demonstrate the presence of a previously unrecognized type of filamentous cross-bridges that link intermediate filaments to the nuclear pore complexes.