Intermittency properties in a temporal lobe epilepsy model.

Neuronal synchronization is important for communication between brain regions and plays a key role in learning. However, changes in connectivity can lead to hyper-synchronized states related to epileptic seizures that occur intermittently with asynchronous states. The activity-regulated cytoskeleton-associated protein (ARC) is related to synaptic alterations which can lead to epilepsy. Induction of status epilepticus in rodent models causes the appearance of intense ARC immunoreactive neurons (IAINs), which present a higher number of connections and conductance intensity than non-IAINs. This alteration might contribute to abnormal epileptic seizure activity. In this work, we investigated how IAINs connectivity influences the firing pattern and synchronization in neural networks. Firstly, we showed the appearance of synchronized burst patterns due to the emergence of IAINs. Second, we described how the increase of IAINs connectivity favors the appearance of intermittent up and down activities associated with synchronous bursts and asynchronous spikes, respectively. Once the intermittent activity was properly characterized, we applied the optogenetics control of the high synchronous activities in the intermittent regime. To do this, we considered that 1% of neurons were transfected and became photosensitive. We observed that optogenetics methods to control synchronized burst patterns are effective when IAINs are chosen as photosensitive, but not effective in non-IAINs. Therefore, our analyses suggest that IAINs play a pivotal role in both the generation and suppression of highly synchronized activities.

Maturação in vitro de oócitos de cutias (Dasyprocta prymnolopha, Wagler 1831) seguida de fertilização in vitro e ativação partenogenética / In vitro maturation of agoutis (Dasyprocta prymnolopha) oocytes followed by in vitro fertilization and parthenogenetic activation

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.(AU)

The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.(AU)

Potencial inseticida de óleos essenciais sobre Tribolium castaneum em milho armazenado / Insecticidal potential of essential oils on Tribolium castaneum in stored maize

RESUMO O presente trabalho teve como objetivo investigar o potencial inseticida de óleos essenciais de Croton heliotropiifolius, Croton pulegiodorus, Myracrodruon urundeuva e Ocimumbasilicum sobre adultos de Tribolium castaneumem milho armazenado. Para cada óleo foram realizados bioensaios de fumigação, repelência e o efeito sobre a taxa instantânea de crescimento (ri), em cinco concentrações (0; 5; 10; 15 e 20 μL). Os bioensaios foram conduzidos sob condições constantes de temperatura (28±2 ºC), umidade relativa (70±5%) e escoto fase de 24 horas. Nos testes de fumigação diferentes concentrações dos óleos foi aplicado em tiras de papel filtro presas na parte inferior da tampa da câmara de fumigação (1,5L), a qual continha 20 gramas de substrato alimentar e 10 insetos adultos de T. castaneum não sexados. A mortalidade dos insetos foi avaliada após 48 horas de exposição. Os testes de repelência foram efetuados em arenas compostas por dois frascos ligados a uma caixa central. Em um frasco foi depositado o substrato alimentar com diferentes concentrações do óleo essencial, e, no outro, foi depositado apenas alimento (testemunha). Dez insetos adultos foram liberados na caixa central, ficando expostos por 5 dias para avaliação da preferência. Nos bioensaios de fumigação observou-se atividade inseticida do óleo essencial de M. urundeuva sobre adultos de T. castaneum. Nos bioensaios de repelência, todos os óleos testados apresentaram efeito repelente. A emergência de T. castaneumreduziu entre 33 e 100% quando foram criados em pó de milho tratado com os óleos essenciais. Os óleos essenciais de C. pulegiodorus e O. basilicum ocasionaram redução do crescimento populacional de T. castaneum em grãos de milho tratados. Os óleos testados demonstraram ser uma alternativa eficiente de controle para o uso nos programas de manejo de T. castaneum em unidades armazenadoras.

ABSTRACT The current work aimed at to investigate the insecticide potential of essential oils of Croton heliotropiifolius, Croton pulegiodorus, Myracrodruon urundeuva and Ocimum basilicum on adults of Tribolium castaneum in stored maize. For each oil, it were performed fumigation tests, repellency and the effect on the instantaneous rate of increase (ri), in five concentrations (0; 5; 10; 15 e 20 μL). Bioassays were carried out under constant temperature conditions (28±2 °C), relative humidity (70±5%) and scot phase of 24 hours. In fumigation different concentrations of test oils were applied on filter paper strips attached on the bottom of the fumigation chamber cover (1.5 L), which contained 20 grams of food substrate and 10 unsexed adults of T. castaneum. The insect mortality was recorded after 48 hours of exposure. The repellency tests were performed in arenas composed of two pots connected to a central box. In a pot it was deposited feed substrate, with concentrations of the essential oil, in the other pot (control) it was deposited only food. Ten adult insects were released in the central box, being exposed for 5 days to evaluate the preference. In the fumigation tests, insecticide activity of the essential oil of M. urundeuva was observed on adults of T. castaneum. In the repellency tests, all the tested oils presented repellent effect. The emergency of T. castaneumreduced between 33% and 100% when they were in the powdered maize treated with the essential oils. Maize grains treated with C. pulegiodorus and O. basilicum essential oils caused a significant decrease in the populations of T. castaneum. The tested oils proved to be an efficient control alternative for the use in managing programs of T. castaneum in storing units.

Encapsulation of trans-dehydrocrotonin in liposomes: an enhancement of the antitumor activity.

The aim of this study was the encapsulation of trans-dehydrocrotonin (t-DCTN) and its inclusion complexes with hydropropyl-beta-cyclodextrin (HP-beta-CD) in liposomes to improve t-DCTN antitumor activity. The in vitro kinetic profiles of t-DCTN-loaded liposomes (LD) and t-DCTN:HP-beta-CD-loaded liposomes (LC) were evaluated using the dialysis technique. The antitumor activity of LD and LC were investigated against Sarcoma 180 in Swiss mice. Histopathological and hematological analyses were carried out. The amounts of t-DCTN and t-DCTN:HP-beta-CD inclusion complex encapsulated in liposomes were equivalent to 1 mg of t-DCTN. The encapsulation efficiencies of LD and LC were 95.0 +/- 3.8% and 91.1 +/- 5.6%, respectively. In relation to kinetics, the drug release profiles of t-DCTN are in substantial agreement with the Fickian model. The treatment of animals with LD and LC produced tumor inhibitions of 79.4 +/- 9.6% and 63.5 +/- 5.5%, respectively. The liposomal encapsulation of t-DCTN by entrapment in the phospholipid bilayer increased at twice the antitumor activity. Moreover, the liposomal formulations reduced the hepatotoxicity effect of the drug and no significant hematological toxicity was observed in the treated animals. However, the counting of platelets was slightly decreased. Thus, the results show that the development of liposomal formulations containing t-DCTN or t-DCTN:HP-beta-CD is an important advance for enabling this drug to be use in therapy.

Pregnancy in Hystricomorpha: gestational age and embryonic-fetal development of agouti (Dasyprocta prymnolopha, Wagler 1831) estimated by ultrasonography.

Thirty-one pregnant agoutis, between Days 9 and 103 of gestation (Day 1 = day of detection of sperm in the vaginal smear), underwent B-mode ultrasonography; gestational sac diameter (GSD), crown-rump length (CRL), embryonic-fetal diameter (EFD), and placenta diameter (PD) were measured. There were positive correlations (P < 0.05) between GSD and CRL (r = 0.98), GSD and PD (r = 0.88), CRL and PD (r = 0.86), days of gestation (DG) and CRL (r = 0.85), and DG and PD (r = 0.73). The gestational sac was first observed on Day 14. The embryo was first seen on Day 18 in 9/31 of pregnant agoutis and on Day 22 in 20/31 of pregnant agoutis. Heartbeats were detected from the Day 25 and placentas were observed in 100% of the animals from Day 25. Early limb bud and ossification of the fetal skull were identified on Days 27 (15/31) and 45 (24/31), respectively. Fetal orientation (head and body) was evident from Day 40, the stomach, liver and lungs were identified on Day 50, the kidneys were reliably seen only on Day 55, and the aorta and vena cava were seen on Day 70. The fetal bowel and the urinary bladder were the last structures to be observed (Day 85). Ultrasonography was effective for early pregnancy diagnosis in agouti and for obtaining information on embryonic and fetal structures that could be used to predict gestational age and birth, thereby contributing to their reproductive management in captivity.

Collection and evaluation of epididymal sperm in captive agoutis (Dasyprocta aguti).

The objective was to establish a protocol for the collection and evaluation of epididymal sperm in agoutis. Eight males (1-2 y old) underwent left orchidectomy and epididymal sperma were collected by retrograde flush. Average values were flush volume 32 µL, pH 6.9, sperm concentration 748 x 10(6) sperm/mL, with motility 86.5% and vigor 4.6. Viable sperm were present in all flush samples; 66% of sperm were alive, and 41.9% of sperm responded positively to the hypoosmotic test (using distilled water). There were 21.1% morphologically abnormal sperm, of which 2.0 and 19.1% were primary and secondary defects, respectively. The acrosome was intact in 99.5% of sperm. The sperm head was 4.89 ± 0.41 µm long and 3.13 ± 0.35 µm wide, with an area of 13.01 ± 2.01 µm(2). Midpieces were 5.33 ± 0.44 µm long and 0.98 ± 0.13 wide, sperm tails were 29.91 ± 2.29 µm, and overall sperm length was 40.12 ± 2.44 µm. In conclusion, epididymal sperm collection from agoutis was satisfactory; the collected sperm has the potential to be stored, facilitating development of other reproductive biotechnologies for this species.