Expressão da aldeído desidrogenase (ALDH), STRO-1 e CD44 em células-tronco da polpa de dentes decíduos e permanentes / Expression of aldehyde dehydrogenase (ALDH), STRO-1 and CD44 in pulp stem cells from deciduous and permanent teeth

O objetivo deste estudo foi avaliar a expressão da enzima aldeído desidrogenase (ALDH), assim como dos marcadores de membrana celular STRO-1 e CD44 em células-tronco da polpa de dentes decíduos (SHEDs), permanentes (DPSCs) e fibroblastos da polpa (HDPFs), através da citometria de fluxo e do western blot. Para tanto, as células foram cultivadas em meio DMEM/ HEPES, suplementado com soro fetal bovino a 10%, 100U/mL de penicilina, 100µg/mL de estreptomicina e armazenadas em estufa a 37ºC e 5% de CO2. Para a avaliação dos resultados da citometria de fluxo foram utilizados o teste não paramétrico de Kruskal-Wallis (p ≤ 0,05) e o teste de comparação múltipla de Dunn. As três linhagens de células, quando caracterizadas pelo western blot, expressaram fortemente a ALDH, assim como o STRO-1, ao passo que apresentaram uma fraca expressão para o CD44, não apresentando diferença na intensidade das bandas entre as mesmas. Já na análise por citometria de fluxo, todas as células apresentaram valores percentuais altos para o CD44, sem diferença estatística. Para o marcador STRO-1, os valores foram relativamente baixos para os três tipos celulares, havendo diferença apenas entre SHEDs e HDPFs. Os valores percentuais para a atividade da enzima ALDH foram igualmente baixos, havendo diferença estatisticamente significante entre DPSCs e HDPFs. Os resultados deste estudo sugerem que SHEDs, DPSCs e fibroblastos podem compartilhar algumas características, como a expressão de determinados marcadores genéticos. Da mesma forma, indicam que o uso da ALDH pode ser explorado em pesquisas futuras, para caracterização e seleção das MSCs.

The aim of this study was to evaluate the aldehyde dehydrogenase (ALDH) activity, as well as the expression of STRO-1 and CD44 in pulp stem cells from deciduous (SHEDs) and permanent (DPSCs) teeth, as well in pulp fibroblasts, by flow cytometry and western blot. For this purpose, cells were cultured in DMEM/ HEPES, supplemented with fetal bovine serum at 10%, 100U/ml of penicillin, 100µg/mL streptomycin and stored at 37°C and 5% CO2. To verify differences in the flow cytometry analysis between the three cell types, the nonparametric Kruskal-Wallis test was used (p  0,05), along with the Dunn multiple comparison test. In the western blot test, all the cell lineages strongly expressed ALDH, as well as STRO-1, while they presented a weak expression for CD44. There were no differences in the band intensity between the three cell types for the proteins tested. In the flow cytometry analysis, all cells showed high percentages of CD44, with no statistical difference. For the STRO-1 marker, the values were relatively low for all the three cell types, and differences were observed just between SHEDs and HDPFs. The values observed for the ALDH activity were also lower, with statistically significant difference between DPSCs and HDPFs. The results of this study suggest that SHEDs, DPSCs and pulp fibroblasts may share some important characteristics, such as the expression of certain genetic markers. Likewise, indicate that the use of ALDH activity can be exploited in future research for identification and characterization of MSCs.

Características do sêmen de carneiros Dorper, Santa Inês e sem padrão racial definido, pré e pós-congelação, nos períodos chuvoso e seco / Characteristics of the semen of Dorper, Santa Ines and undefined breed sheep, pre-and post-freezing, in the rainy and dry period

Avaliaram-se as características espermáticas de carneiros Dorper, Santa Inês e sem padrão racial definido, nos períodos chuvoso e seco. Após ser colhido por vagina artificial, o sêmen foi avaliado quanto ao volume, ao turbilhonamento, à motilidade, ao vigor, à morfologia e à concentração, congelado e armazenado em botijão criogênico. Depois de descongelado, foram avaliadas a cinemática espermática, a integridade da membrana plasmática, a integridade do acrossoma e a atividade mitocondrial. Vigor espermático, motilidade total, motilidade progressiva, velocidade em linha reta e defeitos maiores não diferiram entre os períodos chuvoso e seco, porém volume, turbilhonamento, linearidade, retilinearidade e frequência de batimentos de cauda foram mais baixos (P<0,05) no período seco; já concentração espermática e defeitos totais apresentaram valores mais baixos no período chuvoso. Valores de integridade do acrossoma e da membrana plasmática, bem como o potencial de membrana mitocondrial, foram mais baixos (P<0,05) no período seco. Conclui-se que os períodos chuvoso e seco influenciam na qualidade espermática de ovinos criados na região Meio-Norte do Brasil e que esses animais têm uma qualidade espermática superior no período chuvoso, quando, portanto, deve ocorrer a criopreservação. Também se observou que, em relação à qualidade espermática, o melhor grupo de carneiros foi o Santa Inês...

The sperm characteristics of Dorper, Santa Ines and undefined breed ram in the rainy and dry seasons was assessed. After collection with an artificial vagina, the semen was evaluated for volume, turbulence, motility, viability, morphology and concentration, frozen and stored in a cryogenic cylinder. After thawing kinematic sperm, plasma membrane integrity, acrosome integrity and mitochondrial activity were evaluated. Sperm vigor, total motility, progressive motility, straight line speed and larger line defects did not differ between the rainy and dry seasons, however, volume, turbulence, linearity, straightness and frequency of tail beats were lower (P<0.05 ) in the dry period, and sperm concentration and total defects showed lower values during the rainy season. Values for acrosome integrity and plasma membrane and mitochondrial membrane potential were lower (P<0.05) in the dry season. It is concluded that the wet and dry period influence sperm quality in ram raised in the Mid - North region of Brazil, with a higher sperm quality in the rainy season, thus suggesting cryopreservation for that period. Also, the better ram group regarding sperm quality, was the Santa Ines...

Qualidade do corpo lúteo e do embrião de ovelhas Santa Inês superovuladas com FSH associado à rbST / Quality of corpus luteum and embryo of Santa Inês ewes superovulated with FSH associated with rbST

Avaliou-se o efeito da somatotropina bovina recombinante (rbST) sobre a qualidade do corpo lúteo e do embrião e sobre a concentração de progesterona (P4), em ovelhas da raça Santa Inês doadoras de embriões. Foram utilizadas 13 doadoras, distribuídas aleatoriamente em dois grupos, G1 (n=7) e G2 (n=6). Em ambos os grupos, as doadoras foram sincronizadas com implante vaginal por 14 dias, receberam 100µg de PGF2α (Ciosin®) no sétimo dia e foram superovuladas, a partir do 12º dia, com 250UI de FSH (Folltopin-V®) em seis doses decrescentes. No dia da retirada do implante, as doadoras do G2 receberam 125mg de rbST (Boostin®) e foram cobertas por macho Santa Inês a cada 12 horas até o final do estro. No sexto dia após a primeira cobrição, foram avaliadas, por laparoscopia, a resposta superovulatória e a quantidade e a qualidade dos corpos lúteos. Após as avaliações, as colheitas embrionárias foram realizadas por laparotomia. A administração da rbST no protocolo de superovulação promoveu aumento significativo (P<0,05) no número de corpos lúteos do tipo I (9,00 x 5,28) e na qualidade embrionária (4,33 x 2,00).

We evaluate the effect of recombinant bovine somatotropin (rbST) on the quality of the corpus luteum and embryo and on the concentration of progesterone (P4) in donors embryo sheep Santa Ines. 13 donors were used, randomly assigned to two groups, G1 (n=7) and G2 (n=6). In both groups, the donors were synchronized with vaginal implant for 14 days and received 100µg of PGF2α (Ciosin®) on the seventh day and were superovulated, from the 12th day, with 250IU of FSH (Folltopin-V®) in six decreasing doses. On the day of implant removal the donor G2 received 125mg of rbST (Boostin®), and were covered by Santa Inês male every 12 hours until the end of estrus. In the sixth day after the first mating were evaluated by laparoscopy, superovulatory response and the amount and quality of the corpus luteum. After the evaluations, the embryonic harvests were performed by laparotomy. The administration of rbST in superovulation protocol promoted a significant increase (P <0.05) in the number of corpus luteum of type I(9.00 x 5.28) and embryo quality (4.33 x 2.00).

Recombinant vaccines and the development of new vaccine strategies

Vaccines were initially developed on an empirical basis, relying mostly on attenuation or inactivation of pathogens. Advances in immunology, molecular biology, biochemistry, genomics, and proteomics have added new perspectives to the vaccinology field. The use of recombinant proteins allows the targeting of immune responses focused against few protective antigens. There are a variety of expression systems with different advantages, allowing the production of large quantities of proteins depending on the required characteristics. Live recombinant bacteria or viral vectors effectively stimulate the immune system as in natural infections and have intrinsic adjuvant properties. DNA vaccines, which consist of non-replicating plasmids, can induce strong long-term cellular immune responses. Prime-boost strategies combine different antigen delivery systems to broaden the immune response. In general, all of these strategies have shown advantages and disadvantages, and their use will depend on the knowledge of the mechanisms of infection of the target pathogen and of the immune response required for protection. In this review, we discuss some of the major breakthroughs that have been achieved using recombinant vaccine technologies, as well as new approaches and strategies for vaccine development, including potential shortcomings and risks.

Psychopharmacotherapy of panic disorder: 8-week randomized trial with clonazepam and paroxetine

The objective of the present randomized, open-label, naturalistic 8-week study was to compare the efficacy and safety of treatment with clonazepam (N = 63) and paroxetine (N = 57) in patients with panic disorder with or without agoraphobia. Efficacy assessment included number of panic attacks and clinician ratings of the global severity of panic disorders with the clinical global impression (CGI) improvement (CGI-I) and CGI severity (CGI-S) scales. Most patients were females (69.8 and 68.4 percent in the clonazepam and paroxetine groups, respectively) and age (mean ± SD) was 35.9 ± 9.6 years for the clonazepam group and 33.7 ± 8.8 years for the paroxetine group. Treatment with clonazepam versus paroxetine resulted in fewer weekly panic attacks at week 4 (0.1 vs 0.5, respectively; P < 0.01), and greater clinical improvements at week 8 (CGI-I: 1.6 vs 2.9; P = 0.04). Anxiety severity was significantly reduced with clonazepam versus paroxetine at weeks 1 and 2, with no difference in panic disorder severity. Patients treated with clonazepam had fewer adverse events than patients treated with paroxetine (73 vs 95 percent; P = 0.001). The most common adverse events were drowsiness/fatigue (57 percent), memory/concentration difficulties (24 percent), and sexual dysfunction (11 percent) in the clonazepam group and drowsiness/fatigue (81 percent), sexual dysfunction (70 percent), and nausea/vomiting (61 percent) in the paroxetine group. This naturalistic study confirms the efficacy and tolerability of clonazepam and paroxetine in the acute treatment of patients with panic disorder.

NMDA receptor blockade alters the intracellular distribution of neuronal nitric oxide synthase in the superficial layers of the rat superior colliculus

Nitric oxide (NO) is a molecular messenger involved in several events of synaptic plasticity in the central nervous system. Ca2+ influx through the N-methyl-D-aspartate receptor (NMDAR) triggers the synthesis of NO by activating the enzyme neuronal nitric oxide synthase (nNOS) in postsynaptic densities. Therefore, NMDAR and nNOS are part of the intricate scenario of postsynaptic densities. In the present study, we hypothesized that the intracellular distribution of nNOS in the neurons of superior colliculus (SC) superficial layers is an NMDAR activity-dependent process. We used osmotic minipumps to promote chronic blockade of the receptors with the pharmacological agent MK-801 in the SC of 7 adult rats. The effective blockade of NMDAR was assessed by changes in the protein level of the immediate early gene NGFI-A, which is a well-known NMDAR activity-dependent expressing transcription factor. Upon chronic infusion of MK-801, a decrease of 47 percent in the number of cells expressing NGFI-A was observed in the SC of treated animals. Additionally, the filled dendritic extent by the histochemical product of nicotinamide adenine di-nucleotide phosphate diaphorase was reduced by 45 percent when compared to the contralateral SC of the same animals and by 64 percent when compared to the SC of control animals. We conclude that the proper intracellular localization of nNOS in the retinorecipient layers of SC depends on NMDAR activation. These results are consistent with the view that the participation of NO in the physiological and plastic events of the central nervous system might be closely related to an NMDAR activity-dependent function.

Colony membership is reflected by variations in cuticular hydrocarbon profile in a Neotropical paper wasp, Polistes satan (Hymenoptera, Vespidae)

Nestmate recognition is one the most important features in social insect colonies. Although epicuticular lipids or cuticular hydrocarbons have both structural and defensive functions in insects, they also seem to be involved in several aspects of communication in wasps, bees and ants. We analyzed and described for the first time the cuticular hydrocarbons of a Neotropical paper wasp, Polistes satan, and found that variation in hydrocarbon profile was sufficiently strong to discriminate individuals according to their colony membership. Therefore, it seems that small differences in the proportion of these compounds can be detected and used as a chemical-based cue by nestmates to detect invaders and avoid usurpation.

Inheritance of parthenocarpy in summer squash (Cucurbita pepo L)

The inheritance of the tendency to set parthenocarpic fruit in the summer squash (Cucurbita pepo L.) line Whitaker was studied. Two parental lines, Whitaker (parthenocarpic) and Caserta (non-parthenocarpic), and the F1 and F2 generations and backcrosses to both parents were tested. The parthenocarpic tendency of individual plants was scored on a scale from 1 (non-parthenocarpic fruit) to 5 (parthenocarpic fruit). The Whitaker line produced parthenocarpic fruit and had a mean score of 4.2, whereas Caserta did not set parthenocarpic fruit and had a score of 1.55. The heritability estimates indicated that genetic gains from selection were feasible. The additive-dominant model showed a good fit, with epistasis being negligible or nonexistent. The hypothesis of monogenic inheritance with incomplete dominance was not rejected within the degree of dominance range from 0.2 to 0.5. These results indicate that parthenocarpy is controlled by a single locus, with incomplete dominance in the direction of parthenocarpic expression

Clinical features of panic patients sensitive to hyperventilation or breath-holding methods for inducing panic attacks

Our aim was to compare the clinical features of panic disorder (PD) patients sensitive to hyperventilation or breath-holding methods of inducing panic attacks. Eighty-five PD patients were submitted to both a hyperventilation challenge test and a breath-holding test. They were asked to hyperventilate (30 breaths/min) for 4 min and a week later to hold their breath for as long as possible, four times with a 2-min interval. Anxiety scales were applied before and after the tests. We selected the patients who responded with a panic attack to just one of the tests, i.e., those who had a panic attack after hyperventilating (HPA, N = 24, 16 females, 8 males, mean age ± SD = 38.5 ± 12.7 years) and those who had a panic attack after breath holding (BHPA, N = 20, 11 females, 9 males, mean age ± SD = 42.1 ± 10.6 years). Both groups had similar (chi² = 1.28, d.f. = 1, P = 0.672) respiratory symptoms (fear of dying, chest/pain disconfort, shortness of breath, paresthesias, and feelings of choking) during a panic attack. The criteria of Briggs et al. [British Journal of Psychiatry, 1993; 163: 201-209] for respiratory PD subtype were fulfilled by 18 (75.0 percent) HPA patients and by 14 (70.0 percent) BHPA patients. The HPA group had a later onset of the disease compared to BHPA patients (37.9 ± 11.0 vs 21.3 ± 12.9 years old, Mann-Whitney, P < 0.001), and had a higher family prevalence of PD (70.8 vs 25.0 percent, chi² = 19.65, d.f. = 1, P = 0.041). Our data suggest that these two groups - HPA and BHPA patients - may be specific subtypes of PD.

Smoking and psychiatric disorders: a comorbidity survey

Epidemiological and clinical studies have shown a positive correlation between smoking and psychiatric disorders. To investigate the prevalence of cigarette smoking, 277 psychiatric outpatients with anxiety or depressive disorders (DSM-IV) answered a self-evaluation questionnaire about smoking behavior and were compared with a group of 68 control subjects. The diagnoses (N = 262) were: 30.2 percent (N = 79) major depressive disorder, 23.3 percent (N = 61) panic disorder, 15.6 percent (N = 41) social anxiety disorder, 7.3 percent (N = 19) other anxiety disorders, and 23.7 percent (N = 62) comorbidity disorders. Among them, 26.3 percent (N = 69) were smokers, 23.7 percent (N = 62) were former smokers and 50.0 percent (N = 131) were nonsmokers. The prevalence of nicotine dependence among the smokers was 59.0 percent (DSM-IV). The frequency of cigarette smoking did not show any significant difference among the five classes of diagnosis. The social anxiety disorder patients were the heaviest smokers (75.0 percent), with more unsuccessful attempts to stop smoking (89.0 percent). The frequency of former smokers was significantly higher among older subjects and nonsmokers were significantly younger (chi² = 9.13, d.f. = 2, P = 0.01). Our data present some clinical implications suggesting that in our psychiatric outpatient sample with anxiety disorder, major depression and comorbidity (anxiety disorder and major depression), the frequency of cigarette smoking did not differ from the frequency found in the control group or in general population studies. Some specific features of our population (outpatients, anxiety and depressive disorders) might be responsible for these results

Respiratory panic disorder subtype and sensitivity to the carbon dioxide challenge test

The aim of the present study was to verify the sensitivity to the carbon dioxide (CO2) challenge test of panic disorder (PD) patients with respiratory and nonrespiratory subtypes of the disorder. Our hypothesis is that the respiratory subtype is more sensitive to 35 percent CO2. Twenty-seven PD subjects with or without agoraphobia were classified into respiratory and nonrespiratory subtypes on the basis of the presence of respiratory symptoms during their panic attacks. The tests were carried out in a double-blind manner using two mixtures: 1) 35 percent CO2 and 65 percent O2, and 2) 100 percent atmospheric compressed air, 20 min apart. The tests were repeated after 2 weeks during which the participants in the study did not receive any psychotropic drugs. At least 15 of 16 (93.7 percent) respiratory PD subtype patients and 5 of 11 (43.4 percent) nonrespiratory PD patients had a panic attack during one of two CO2 challenges (P = 0.009, Fisher exact test). Respiratory PD subtype patients were more sensitive to the CO2 challenge test. There was agreement between the severity of PD measured by the Clinical Global Impression (CGI) Scale and the subtype of PD. Higher CGI scores in the respiratory PD subtype could reflect a greater sensitivity to the CO2 challenge due to a greater severity of PD. Carbon dioxide challenges in PD may define PD subtypes and their underlying mechanisms

Hyperventilation in panic disorder patients and healthy first-degree relatives

Our aim was to observe the induction of panic attacks by a hyperventilation challenge test in panic disorder patients (DSM-IV) and their healthy first-degree relatives. We randomly selected 25 panic disorder patients, 31 healthy first-degree relatives of probands with panic disorder and 26 normal volunteers with no family history of panic disorder. All patients had no psychotropic drugs for at least one week. They were induced to hyperventilate (30 breaths/min) for 4 min and anxiety scales were applied before and after the test. A total of 44.0 percent (N = 11) panic disorder patients, 16.1 percent (N = 5) of first-degree relatives and 11.5 percent (N = 3) of control subjects had a panic attack after hyperventilating (chi2 = 8.93, d.f. = 2, P = 0.011). In this challenge test the panic disorder patients were more sensitive to hyperventilation than first-degree relatives and normal volunteers. Although the hyperventilation test has a low sensitivity, our data suggest that there is no association between a family history of panic disorder and hyperreactivity to an acute hyperventilation challenge test. Perhaps cognitive variables should be considered to play a specific role in this association since symptoms of a panic attack and acute hyperventilation overlap

Análise da monitorizaçäo ambulatorial de pressäo arterial em hipertensos leves / Analysis of ambulatory blood pressure monitoring in mild hypertensive patients

Os autores analisam a curva pressórica obtida pela Monitorizaçäo Ambulatorial da Pressäo Arterial (M. A. P. A) em 15 pacientes hipertensos primários brancos, forma leve, sem comprometimento de orgäos alvos, e observam que os valores tencionais säo inferiores àqueles observados pela aferiçäo casual. Sugerem que a variabilidade e a carga pressórica possam ser fatores preditivos e prognósticos da doença hipertensiva e mostram a importância da ascensäo matinal, demonstrando que o horário de pico é entr6 e 7 horas, e demonstram que o incremento do duplo produto observado neste período é preponderamente devido ao aumento da frequência cardíaca do que ao aumento da pressäo arterial.

Miopatia miotubular: estudo clinico, etrophysiological and histological study so. / Myotubular myopathy: clinical,elecof a case

Sao apresentados os resultados dos exames clinicos, eletrofisiologicos e do estudo do ponto motor, da imunofluorescencia, da histoquimica e da ultramicroscopia da biopsia muscular de um caso de miopatia miotubular. Sao discutidos estes resultados em relacao aos achados de 56 casos desta molestia consignados na literatura ate 1978, sendo cada enfase a etiopatogenia