Reproductive aspects of the flyingfish, Hirundichthys affinis from the Northeastern coastal waters of Brazil.

The epipelagic flyingfish, Hirundichthys affinis is a major artisanal fishery resource from the Northeastern coastal waters of Brazil. However, biological information about this species has been poorly documented. This paper presents data on the length-weight relationship, sex ratio, length at first sexual maturity, gonadal development and fecundity of H. affinis sampled from the coastal waters of Rio Grande do Norte, Brazil. The total body length and weight for both sexes ranged from 23.4 to 29.4 cm and from 89 to 188g, respectively. The allometric coefficient of males was 2.208 and that of females was 2.985, indicating negatively allometric growth. The sex ratio was 1M:1.6F thus differing from the expected ratio of 1:1 (χ2 = 18.63). The total length at first sexual maturity was estimated at 27.3 cm for males and 27.1 cm for females. The macroscopic characteristics of the gonads indicated four maturation stages. Histological studies of gonads of H. affinis showed seven phases of oocyte development and four phases of spermatocyte development. The mean absolute fecundity was 9092 vitelogenic oocytes. Spawning occurred during the months of March to July. The microscopic descriptions of the stages of gonad maturation indicate that the study area is an important spawning ground of H. affinis.

Animal model for chronic massive rotator cuff tear: behavioural and histologic analysis.

PURPOSE: Massive rotator cuff tears (MRCT) are usually chronic lesions that present associated degenerative changes of the myotendinous unit that have been implicated in limitations for surgical repair. In order to develop effective therapies, it is important to establish animal models that mimic the hallmarks of the injury itself. Therefore, in the present work, we aimed to (1) optimize a rodent animal model of MRCT that closely reproduces the fatty infiltration of the cuff muscles seen in humans and (2) describe the effects of unilateral or bilateral lesion in terms of histology and behaviour. METHODS: Massive tear was defined as two rotator cuff tendons-supraspinatus and infraspinatus-section. Twenty-one Wistar rats were randomly assigned to four groups: bilateral lesion (five animals), right-sided unilateral lesion (five animals), left-sided unilateral lesion (five animals) and control (six animals). Behaviour was analyzed with open field and staircase test, 16 weeks after lesion. After that, animals were killed, and the supraspinatus and infraspinatus muscles were processed. RESULTS: Histologic analysis revealed adipocytes, fatty infiltration and atrophy in the injured side with a greater consistency of these degenerative changes in the bilateral lesion group. Behaviour analysis revealed a significant functional impairment of the fine motor control of the forepaw analyzed in staircase test where the number of eaten pellets was significantly higher in sham animals (sham = 7 ± 5.0; left unilateral = 2.6 ± 3.0; right unilateral = 0 ± 0; and bilateral = 0 ± 0, p < 0.05). A trend to reach a lower level of steps, in more injured animals, was also observed (sham animals = 3 ± 1.6 > left unilateral = 2 ± 2.1 > right unilateral = 0.8 ± 1.3 > bilateral = 0.8 ± 1.1). CONCLUSIONS: The present study has been able to establish an animal model that disclosed the hallmarks of MRCT. This can now be used as a valuable, cost-effective, pre-clinical instrument to assist in the development of advanced tissue engineered strategies. Moreover, this animal model overcomes some of the limitations of those that have been reported so far and thus represents a more reliable source for the assessment of future therapeutic strategies with potential clinical relevance.

The motivational drive to natural rewards is modulated by prenatal glucocorticoid exposure.

Exposure to elevated levels of glucocorticoids (GCs) during neurodevelopment has been identified as a triggering factor for the development of reward-associated disorders in adulthood. Disturbances in the neural networks responsible for the complex processes that assign value to rewards and associated stimuli are critical for disorders such as depression, obsessive­compulsive disorders, obesity and addiction. Essential in the understanding on how cues influence behavior is the Pavlovian­instrumental transfer (PIT), a phenomenon that refers to the capacity of a Pavlovian stimulus that predicts a reward to elicit instrumental responses for that same reward. Here, we demonstrate that in utero exposure to GCs (iuGC) impairs both general and selective versions of the PIT paradigm, suggestive of deficits in motivational drive. The iuGC animals presented impaired neuronal activation pattern upon PIT performance in cortical and limbic regions, as well as morphometric changes and reduced levels of dopamine in prefrontal and orbitofrontal cortices, key regions involved in the integration of Pavlovian and instrumental stimuli. Normalization of dopamine levels rescued this behavior, a process that relied on D2/D3, but not D1, dopamine receptor activation. In summary, iuGC exposure programs the mesocorticolimbic dopaminergic circuitry, leading to a reduction in the attribution of the incentive salience to cues, in a dopamine-D2/D3-dependent manner. Ultimately, these results are important to understand how GCs bias incentive processes, a fact that is particularly relevant for disorders where differential attribution of incentive salience is critical.

Mechanisms of initiation and reversal of drug-seeking behavior induced by prenatal exposure to glucocorticoids.

Stress and exposure to glucocorticoids (GC) during early life render individuals vulnerable to brain disorders by inducing structural and chemical alterations in specific neural substrates. Here we show that adult rats that had been exposed to in utero GCs (iuGC) display increased preference for opiates and ethanol, and are more responsive to the psychostimulatory actions of morphine. These animals presented prominent changes in the nucleus accumbens (NAcc), a key component of the mesolimbic reward circuitry; specifically, cell numbers and dopamine (DA) levels were significantly reduced, whereas DA receptor 2 (Drd2) mRNA expression levels were markedly upregulated in the NAcc. Interestingly, repeated morphine exposure significantly downregulated Drd2 expression in iuGC-exposed animals, in parallel with increased DNA methylation of the Drd2 gene. Administration of a therapeutic dose of L-dopa reverted the hypodopaminergic state in the NAcc of iuGC animals, normalized Drd2 expression and prevented morphine-induced hypermethylation of the Drd2 promoter. In addition, L-dopa treatment promoted dendritic and synaptic plasticity in the NAcc and, importantly, reversed drug-seeking behavior. These results reveal a new mechanism through which drug-seeking behaviors may emerge and suggest that a brief and simple pharmacological intervention can restrain these behaviors in vulnerable individuals.

Perfil do conhecimento de cuidadores de pacientes pediátricos sobre medicamentos prescritos / Profile of the knowledge of pediatric patient caregivers about prescribed medicines

O raro contato entre os profissionais de saúde e o cuidador do paciente pediátrico, associado ao pouco tempo disponibilizado pelo profissional farmacêutico durante a dispensação, não são suficientes para a geração de informações significativas, podendo acarretar falhas no fluxo de informação e não adesão involuntária do paciente ao tratamento. O objetivo do trabalho foi coletar dados sobre o perfil do conhecimento dos cuidadores de pacientes pediátricos a respeito de medicamentos dispensados na Farmácia Ambulatorial do Hospital Infantil Albert Sabin (HIAS), com enfoque para os medicamentos do Componente Especializado da Assistência Farmacêutica nas especialidades de Neurologia e Pneumologia. Trata-se de um estudo prospectivo. A coleta de dados ocorreu entre novembro de 2007 e maio de 2008, utilizando-se um questionário semiestruturado. De acordo com os dados coletados, 87,7% (n=50) eram do gênero feminino; o grau de escolaridade, predominante, foi o ensino médio completo; a renda média familiar ficou compreendida entre um e dois salários mínimos, equivalente a 63,1% dos casos. As questões sobre informações sobre os medicamentos dispensados configuraram-se dessa forma: 12% desconhecem o nome do medicamento prescrito; 14% desconhecem a indicação do medicamento; 7% desconhecem como administrá-lo; 29,8% desconhecem o tempo de tratamento; 82,5% desconhecem o que é reação adversa a medicamento. Foram consideradas como respostas insatisfatórias aquelas que não foram condizentes com as informações contidas nas bulas e/ou receituários médicos dos medicamentos. A análise dos dados permitiu verificar que há, em relação à falta de informação dos entrevistados, dados significantes que podem comprometer a terapia, prejudicando a saúde do paciente.

The rare contacts between health professionals and adult caregivers of pediatric patients in a hospital outpatient pharmacy, together with the short time available during the dispensing, are insufficient to generate a meaningful flow of information, and can lead to an unintentional lack of patient compliance to the treatment. The objective of the study was to collect data on the profile of knowledge of pediatric patient caregivers, regarding medicines dispensed in the outpatient pharmacy at the Hospital Infantil Albert Sabin (a childrens hospital), focusing on those who obtain medicines from the Specialized Pharmaceutical Care component of the Public Health Service, in the specialties of Neurology and Pulmonology. This prospective study was based on data collected from November 2007 to May 2008, using a semi-structured questionnaire. According to the results, 87.7% of the caregivers (n = 50) were female; they were predominantly educated to high school level; the median family income was between one and two times the ?minimum wage? (inflation-adjusted official subsistence wage), 63.1% of incomes falling within this range(?). Questions of fact about the medication dispensed elicited the following profile of respondents? knowledge: 12% did not know the name of the drug, 14% the reason for taking the drug, 7% how to administer the medicine, 29.8% the treatment time and 82.5% did not know about adverse reactions to the medication. Answers considered unsatisfactory were those that were inconsistent with the information contained in the package inserts and / or medical prescriptions of the drugs. Data analysis showed that there was a significant lack of information among the respondents that could impair the adherence to drug therapy and affect patient health.

Strain-dependent arthritogenic potential of the zoonotic pathogen Corynebacterium ulcerans.

During the last decade the majority of diphtheria cases in Europe had Corynebacterium ulcerans as the etiologic agent with dogs and cats as the reservoir hosts. However, little has been documented about the virulence factors of this zoonotic pathogen. To set up an in vivo experimental C. ulcerans infection model, conventional Swiss Webster mice were intravenously infected with different doses (from 1 × 10(7) to 5 × 10(9) bacteria per mouse) of C. ulcerans strains, namely 809 (from human lower respiratory tract), BR-AD22 (from asymptomatic dog nares) and CDC-KC279. Mortality rates were demonstrated by LD(50) values ranging from 1.9 × 10(8) to 1.3 × 10(9). Viable bacteria were recovered from blood, kidneys, liver, spleen and joints. For CDC-KC279 and 809 strains (2 × 10(8)mL(-1)) approximately 85% and 72% of animals with articular lesions were observed, respectively; BR-AD22-infected mice showed no signs of arthritis. CDC-KC279 and 809 strains exhibited higher arthritogenic potential when compared to the homologous toxigenic (ATCC27012) and non-toxigenic (ATCC27010) strains of Corynebacterium diphtheriae. A high number of affected joints and arthritis index in addition to the histopathological features, including subcutaneous edema, inflammatory infiltrate, damage to bone tissue and synoviocyte hypertrophy, indicated a strain-dependent ability of C. ulcerans strains to cause severe polyarthritis. A correlation between the arthritis index and systemic levels of IL-6 and TNF-α was observed for C. ulcerans strains, with the exception of the non-arthritogenic BR-AD22 strain. In conclusion, C. ulcerans revealed a strain-dependent arthritogenic potential independent of DNAse, PLD and diphtheria toxin production.

Triiodothyronine modulates differential homing of recent thymic emigrants to peripheral lymphoid organs.

The functioning of the immune system partially relies on T-cell exportation from the thymus, the major site of T-cell differentiation. Although the molecular mechanisms governing this process begin to be elucidated, it is not clear if thyroid hormones can alter the homing of recent thymic emigrants (RTE) to peripheral lymphoid organs. Herein, we investigated whether triiodothyronine (T(3)) could influence the homing of thymus-derived T cells. For that we used intrathymic injection of T(3) in combination with fluorescein isothiocyanate (FITC) to trace, 16 h later, FITC(+) cells, termed RTE, in peripheral lymphoid organs. We observed that T(3) stimulated thymocyte export, increasing the frequency of CD4(+) RTE and CD8(+) RTE in the subcutaneous and mesenteric lymph nodes. By contrast, the relative numbers of CD4(+) RTE in the spleen were decreased. T(3) also changed the differential distribution pattern of CD4(+) RTE, and to a lesser extent CD8(+) RTE in the peripheral lymphoid organs. Moreover, the expression of extracellular matrix (ECM) components, such as laminin and fibronectin, which are known to be involved in T-cell migration, increased in the lymph nodes but not in the spleen following intrathymic T(3) treatment. In conclusion, our data correspond to the first demonstration that in vivo treatment with thyroid hormone stimulates thymic T-cell homing and T-cell distribution in peripheral lymphoid organs.

Triiodothyronine modulates thymocyte migration.

Triiodothyronine (T(3)) exerts several effects on thymus physiology. In this sense, T(3) is known to stimulate thymic microenvironmental cells to enhance the production of extracellular matrix (ECM) moieties, which are relevant in thymocyte migration. Here, we further investigated the in vivo influence of T(3) on ECM production, as well as on ECM-related T-cell migration events. For this, BALB/c mice were subjected to two protocols of T(3) treatment: long-term (30 days) i.p. daily T(3) injections or short-term (16 h) after a single T(3) intrathymic injection. These two treatments did promote an enhancement in the expression of fibronectin and laminin, in both cortex and medullary regions of the thymic lobules. As revealed by the long-term treatment, the expression of ECM protein receptors, including VLA-4, VLA-5 and VLA-6, was also increased in thymocyte subsets issued from T(3)-treated mice. We further used thymic nurse cells (TNC) as an in vitro system to study the ECM-related migration of immature thymocytes in the context of thymic epithelial cells. Even a single intrathymic injection of T(3) resulted in an increase in the ex vivo exit of thymocytes from TNC lymphoepithelial complexes. Accordingly, when we evaluated thymocyte migration in transwell chambers pre-coated with ECM proteins, we found an increase in the numbers of migrating cells, when thymocytes were derived from T(3)-treated mice. Overall, our data show that in vivo intrathymic short-term i.p. long-term T(3) treatments are able to modulate thymocyte migration, probably via ECM-mediated interactions.

Long-term culture of multibacillary leprosy macrophages isolated from skin lesions: a new model to study Mycobacterium leprae-human cell interaction.

BACKGROUND: Leprosy is characterized by a disease spectrum having two polar clinical forms dependent on the presence or not of cell-mediated immunity. In the tuberculoid forms, granuloma-activated macrophages kill Mycobacterium leprae in conjunction with a Th1 response while, in multibacillary (MB) lesions, M. leprae nonactivated macrophages infiltrate the nerves and internal organs together with a Th2 response. The functional properties and activation pathways of macrophages isolated from patients with MB leprosy remain only partially understood. OBJECTIVES: To establish an ex vivo methodology capable of evaluating the activation pathways, grade and fate of cultured macrophages isolated from MB lesions. METHODS: Skin biopsies from patients with borderline tuberculoid, bordeline lepromatous and lepromatous leprosy (LL) were characterized by immunohistochemistry and transcriptional analysis. To isolate inflammatory cells, a portion of the samples was submitted to enzymatic digestion. These same cells, maintained in culture for a minimum 7-day period, were characterized morphologically and via flow cytometry at different culture time points. Cytokine [interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-10] mRNA levels were quantified by real-time polymerase chain reaction and protein secretion in the culture supernatants was measured by enzyme-linked immunosorbent assay and the nitric oxide levels by Griess reagent. RESULTS: RNA expression in tuberculoid and MB lesions showed the profile expected of characteristic Th1 and Th2 responses, respectively. The inflammatory cells in all biopsies were successfully isolated. Although the number of cells varied between biopsies, it was highest in LL biopsies. The frequency of isolated CD14+ and CD3+ cells measured by flow cytometry correlated with the percentages of macrophages and lymphocytes in the lesions. Throughout the culture period, CD68+ macrophages showed morphological changes. A progressive increase in cell number and reduction of infected cells were perceptible in the cultures. In contrast to the biopsies, TNF-alpha, IFN-gamma and IL-10 expression in the tuberculoid and MB leprosy cells in 24-h culture and the cytokine levels in the supernatants did not differ significantly. During the culture period, cytokine expression in the MB cells progressively declined, whereas, from days 1 to 7, nitrite levels progressively increased. After day 40, the remaining macrophages were able to ingest fluorescein isothiocyanate-labelled M. leprae. These data need to be confirmed. CONCLUSIONS: This study confirmed the feasibility of obtaining ex vivo macrophages from leprosy lesions and keeping them in long-term culture. This procedure may open new pathways to studying the interaction between M. leprae and human macrophages, which might, in turn, lead to the development of therapeutic tools capable of overcoming the specific anergy found in patients with MB leprosy.

Detection of in vitro interferon-gamma and serum tumour necrosis factor-alpha in multidrug-resistant tuberculosis patients.

Multidrug-resistant tuberculosis (MDR-TB) is known as having a poor prognosis with a weak response to therapy and very high death rates. The aim of this work was to assess the immune response to the RD1-encoded antigen ESAT-6 of Mycobacterium tuberculosis in MDR-TB patients and compare to non-resistant (NR) TB patients and healthy controls (HC). Evaluation of interferon (IFN)-gamma production showed that, although 55% of the MDR patients were responsive to ESAT-6, they produced lower IFN-gamma levels (553 +/- 11 pg/ml) when compared to NR-TB (1179 +/- 163 pg/ml; P < 0.05) but not to controls (412 +/- 65.7 pg/ml). Differences in the response to ESAT-6 and to its overlapping peptides mixture were also significant between MDR versus treated pulmonary NR-TB. Furthermore, a very low rate of response to PPD (23.5%) and to Ag85B (33.3%) was noted in MDR-TB patients as compared to the other groups. To determine the inflammatory response in patients' groups, detection of tumour necrosis factor (TNF)-alpha was assessed in their sera before and during chemotherapy. Mean TNF-alpha levels in MDR-TB (43.8 +/- 9 pg/ml) paralleled those found in treated pulmonary, and it was significantly different (P < 0.05) from the values found in untreated NR and HC. Interestingly, secretion of IFN-gamma and TNF-alpha were predominant in MDR patients who presented with bilateral pulmonary lesions and lung cavitation. The present data indicate that the overall immune response to mycobacterial antigens is decreased in resistant TB and the major role inflammatory cytokines may play in perpetuating pulmonary tissue damage.

Evaluation of efficacy and safety of itraconazole oral solution for the treatment of oropharyngeal candidiasis in aids patients.

This study was a non-comparative multicenter clinical trial to evaluate the efficacy and tolerability of itraconazole oral solution 200 mg/day (100 mg twice a day in the fasting state) for the treatment of oropharyngeal candidiasis in AIDS patients. We included 50 patients who were treated and followed for up to 3 weeks after ending therapy in the analysis. Mycological cures at the end of therapy occurred in 20/50 patients (40%), but colonization by Candida sp. was recorded in 42/50 (84%) by the end of follow-up. A high rate of clinical response was observed in 46/50 (92%), and the response was sustained for up to 21 days after stopping therapy in 24/46 patients (52%). Clinical relapses were documented among 22 patients, but all causative fungal organisms associated with a relapse were susceptible to itraconazole. There were many patients with persistence or recurrence of Candida, but without mucositis. Relapse of Candida mucositis was significantly related to low levels of CD(4) lymphocytes exhibited by symptomatic patients. The drug was well tolerated by all but 1 patient. We conclude that itraconazole oral solution (100 mg bid for 7-14 days) is a well tolerated and effective treatment for suppressing the symptoms of oropharyngeal candidiasis in AIDS patients. Patients with severe immunosuppression may relapse and require frequent cycles of treatment or longterm suppressive therapy.

Diet and medication in the treatment of hyperuricemia in hypertensive patients.

OBJECTIVE: To evaluate the effects of diet and medication, either isolated or associated, on serum levels of uric acid in patients with hyperuricemia. METHODS: We studied patients from the Hypertension Unit of the University of Goias who had hyperuricemia (men > or =8.5 mg/dL and women > or =7.5 mg/dL). We divided the patients into three groups: G1 (low purine diet), G2 (low purine diet + medication), and G3 (medication only). Patients received allopurinol, 150 mg/day titrated up to 300 mg/dL when necessary. Patients were evaluated with regards to their lifestyles (diet, smoking, physical, activity, alcohol consumption), uric acid, blood pressure, use of medication, body mass index, cholesterol, and triglyceride. Follow-up took place in weeks 0 (M1), 6 (M2), 12 (M3) during the intervention and in week 36 (M4) after the study was completed. RESULTS: Fifty-five patients participated in the study, 31 women, mean age 54.4+/-10.6 years, body mass index 28.6+/-3.9 kg/m2. A similar reduction (p<0.001) in uric acid levels occurred in the three intervention groups. In week 36 (M4), after 24 weeks without intervention, a tendency toward elevation of uricemia was noted in G2 and G3, and a continuous drop in uricemia was noted in G1. No significant modifications were observed in the other variables analyzed. CONCLUSION: Considering the cost x benefit relationship, a diet low in purine should be the 1st therapeutic option for controlling hyperuricemia in patients with similar characteristic to the ones presented in this study.

Fatores de crescimento: novas perspectivas para regeneração periodontal / The growth factors: the news perspectivas of the periodontal regeneration

Uma série de terapias já foram tentadas visando a completa regeneração dos tecidos periodontais perdidos durante o processo inflamatório, obtendo diferentes graus de regeneração: enxertos ósseos e de outros materiais; condicionamento ácido da superfície radicular; barreiras e, uma sugestão mais recente, aplicação de fatores de crescimento. Dentre os fatores de crescimento já caracterizados, cinco deles parecem muito promissores no campo da regeneração periodontal: fator de crescimento derivado de plaquetas (PDGF); fator de crescimento semelhante à insulina (IGF); fator de crescimento transformador-b (TGF-b), fator de crescimento dos fibroplastos (FGF) e proteínas ósseas morfogenéticas (BMP), pois são capazes de estimular a proliferação de células com fenótipo osteoblástico ou do ligamento periodontal, promover a formação óssea ou a regeneração periodontal em estudos em animais ou in vitro. Baseados nas evidências da ação que estes fatores de crescimento exercem sobre as células responsáveis pela regeneração tecidual, concluímos que seu uso na terapia periodontal é bastente promissor, necessitando apenas de pesquisas mais extensas, Para garantir a segurança de sua utilização em seres humanos

The complete amino acid sequence of the major Kunitz trypsin inhibitor from the seeds of Prosopsis juliflora.

The major inhibitor of trypsin in seeds of Prosopsis juliflora was purified by precipitation with ammonium sulphate, ion-exchange column chromatography on DEAE- and CM-Sepharose and preparative reverse phase HPLC on a Vydac C-18 column. The protein inhibited trypsin in the stoichiometric ratio of 1:1, but had only weak activity against chymotrypsin and did not inhibit human salivary or porcine pancreatic alpha-amylases. SDS-PAGE indicated that the inhibitor has a Mr of ca 20,000, and IEF-PAGE showed that the pI is 8.8. The complete amino acid sequence was determined by automatic degradation, and by DABITC/PITC microsequence analysis of peptides obtained from enzyme digestions of the reduced and S-carboxymethylated protein with trypsin, chymotrypsin, elastase, the Glu-specific protease from S. aureus and the Lys-specific protease from Lysobacter enzymogenes. The inhibitor consisted of two polypeptide chains, of 137 residues (alpha chain) and 38 residues (beta chain) linked together by a single disulphide bond. The amino acid sequence of the protein exhibited homology with a number of Kunitz proteinase inhibitors from other legume seeds, the bifunctional subtilisin/alpha-amylase inhibitors from cereals and the taste-modifying protein miraculin.