Contribuições a elucidação da estrutura, origem e evolução de neo-sistemas cromossômicos de determinação sexual em gafanhotos utilizando como modelo espécies dos gêneros Chlorus, Dichromatos e Eurotettix (Melanoplinae; Acrididae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reproductive efficiency of adult and prepubertal goats subjected to repeated follicular aspiration

The objective of this study was to assess the reproductive response of adult and prepubertal goats subjected to repeated laparoscopic ovum pick-up (LOPU). The study animals were divided into two groups, specifically, adult nanny goats (GA, n=10) and prepubertal nanny goats (GP, n=10), which were subjected to estrous synchronization and ovarian stimulation for LOPU. Both groups underwent six LOPU procedures at seven-day intervals and were subsequently subjected to controlled mating and pregnancy diagnosis to evaluate their future fertility. The study showed a reduction in the number of follicles visualized and in the amount and quality of the oocytes that were recovered and exposed to in vitro maturation. As indicated by the fertility test, however, no complications were found during the laparoscopic procedures that would impair the reproductive future of the animals. Therefore, a viable number of oocytes were obtained even with the decreased reproductive efficiency, proving that repeated LOPUs do not interfere with the reproductive of adult and prepubertal nanny goats. These results indicate a positive aspect of this procedure, allowing for increasing reproductive performance of this kind, when used for the production in vitro.

Prevention of repeated episodes of type 2 reaction of leprosy with the use of thalidomide 100 mg/day

BACKGROUND: Leprosy can have its course interrupted by type 1 and 2 reactional episodes, the last named of erythema nodosum leprosum (ENL). Thalidomide has been the medication of choice for the control of ENL episodes since 1965. OBJECTIVES: These episodes can repeat and cause damages to the patient. In order to prevent these episodes, an extra dose of 100 mg/day thalidomide was used during six months, followed by a follow-up period of six more months after thalidomide discontinuation. METHODS: We included 42 patients with multibacillary (MB) leprosy who had episodes of ENL. They were male and female patients aged between 18 and 84 years. RESULTS: Of the 42 patients, 39 (92.85%) had the lepromatous form and three (7.15%) had the borderline form. We found that 100% of patients had no reactional episode during the use of the drug. During the follow-up period after thalidomide discontinuation, 33 (78.57%) patients had no reactional episode and nine (21.43%), all of them with the lepromatous form, had mild episodes, which were controlled using non-steroidal anti-inflammatory. There were no thalidomide-related side effects. CONCLUSION: A maintenance dose of 100 mg/day of thalidomide showed to be effective to prevent repeated type 2 reactional episodes of ENL.

Influence of Implant Drill Materials on Wear, Deformation, and Roughness After Repeated Drilling and Sterilization

Purpose:The aim of this study was to evaluate deformation, roughness, and mass loss of stainless steel, diamond-like carbon (DLC)-coated and zirconia drills after multiple osteotomies with sterilization procedures.Materials and Methods:Drilling procedures were performed using stainless steel (G1), DLC-coated (G2), and zirconia (G3) drills. All groups were divided in subgroups 1, 2, 3, 4, and 5, corresponded to drills used 0, 10, 20, 30, and 40 times, respectively.Results:No significant differences in mass and roughness were detected among all groups and subgroups. In SEM images, all groups revealed signs of wear while coating delamination was detected in G2. Drills from G1 displayed more irregular surface, whereas cutting edges were more regular in G3.Conclusion:Zirconia drills presented more regular surfaces whereas stainless steel drills revealed more severe signs of wear. Further studies must be performed to evaluate the putative influence of these findings in heat generation.

Organização de DNAs microssatélites nos cromossomos do gafanhoto Abracris flavolineata, com ênfase em cromossomos supranumerários

Com o propósito de se obter um conhecimento aprofundado sobre DNAs repetitivos e sua organização nos cromossomos de gafanhotos, nós utilizamos a técnica citogenética de Hibridização in situ fluorescente (FISH) para o mapeamento da distribuição de dezesseis sequências de microssatélites, incluindo mono-, di-, tri- e tetra-nucleotídeos, nos cromossomos da espécie Abracris flavolineata (Acrididae), que comporta um cromossomo B (supranumerário). A FISH (Hibridização in situ fluorescente) revelou pelo menos dois padrões distintos: (i) sinais exclusivamente espalhados, e (ii) sinais espalhados e específicos, formando blocos evidentes. O enriquecimento foi observado em ambas áreas de eucromatina e heterocromatina e também apenas o motivo (C)30 apresentou ausência na heterocromatina. Os cromossomos A e B se apresentaram enriquecidos com todos os elementos mapeados, sendo observado para o cromossomo a presença de blocos mais distintivos para (GA)15 e (GAG)10. Para o complemento A blocos distintos foram observados para (A)30, (CA)15, (CG)15, (GA)15, (CAC)10, (CAA)10, (CGG)10, (GAA)10, (GAC)10 e (GATA)8. Estes resultados revelaram um intenso espalhamento dos microssatélites no genoma de Abracris flavolineata independentemente de enriquecimento de A+T ou G+C de cada uma das sequências. Os dados indicam que os microssatélites compõem o cromossomo B e que poderiam estar envolvidos na evolução deste elemento na espécie em estudo, embora nenhuma relação específica com qualquer cromossomo A tenha sido observado para discutir sobre sua origem. A análise sistemática apresentada neste trabalho contribui para o conhecimento sobre DNAs repetitivos e sua organização nos cromossomos dos gafanhotos, incluindo os cromossomos B

Repeated sprint ability tests and intensity-time curvature constant to predict short-distance running performances

Anaerobic efforts are commonly required through repeated sprint during efforts in many sports, making the anaerobic pathway a target of training. Nevertheless, to identify improvements on such energetic way it is necessary to assess anaerobic capacity or power, which is usually complex. For this purpose, authors have postulated the use of short running performances to anaerobic ability assessment. Thus, the aim of this study was to find a relationship between running performances on anaerobic power, anaerobic capacity or repeated sprint ability. Methods Thirteen military performed maximal running of 50 (P50), 100 (P100) and 300 (P300) m on track, beyond of running-based anaerobic sprint test (RAST; RSA and anaerobic power test), maximal anaerobic running test (MART; RSA and anaerobic capacity test) and the W′ from critical power model (anaerobic capacity test). Results By RAST variables, peak and average power (absolute and relative) and maximum velocity were significantly correlated with P50 (r = −0.68, p = 0.03 and −0.76, p = 0.01; −0.83, p < 0.01 and −0.83, p < 0.01; and −0.78, p < 0.01), respectively. The maximum intensity of MART was negatively and significantly correlated with P100 (r = −0.59) and W′ was not statistically correlated with any of the performances. Conclusion MART and W′ were not correlated with short running performances, having a weak performance predicting probably due to its longer duration in relation to assessed performances. Observing RAST outcomes, we postulated that such a protocol can be used during daily training as short running performance predictor.

Isolamento de sequências repetitivas do genoma de espécies do gênero Ancistrus (Siluriformes: Loricariidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Duration of the Effects of Repeated Widespread Badger Culling on Cattle Tuberculosis Following the Cessation of Culling

Background: In the British Isles, control of cattle tuberculosis (TB) is hindered by persistent infection of wild badger (Meles meles) populations. A large-scale field trial—the Randomised Badger Culling Trial (RBCT)—previously showed that widespread badger culling produced modest reductions in cattle TB incidence during culling, which were offset by elevated TB risks for cattle on adjoining lands. Once culling was halted, beneficial effects inside culling areas increased, while detrimental effects on adjoining lands disappeared. However, a full assessment of the utility of badger culling requires information on the duration of culling effects. Methodology/Principal Findings: We monitored cattle TB incidence in and around RBCT areas after culling ended. We found that benefits inside culled areas declined over time, and were no longer detectable by three years post-culling. On adjoining lands, a trend suggesting beneficial effects immediately after the end of culling was insignificant, and disappeared after 18 months post-culling. From completion of the first cull to the loss of detectable effects (an average five-year culling period plus 2.5 years post-culling), cattle TB incidence was 28.7% lower (95% confidence interval [CI] 20.7 to 35.8% lower) inside ten 100 km2 culled areas than inside ten matched no-culling areas, and comparable (11.7% higher, 95% CI: 13.0% lower to 43.4% higher, p = 0.39) on lands #2 km outside culled and no-culling areas. The financial costs of culling an idealized 150 km2 area would exceed the savings achieved through reduced cattle TB, by factors of 2 to 3.5. Conclusions/Significance: Our findings show that the reductions in cattle TB incidence achieved by repeated badger culling were not sustained in the long term after culling ended and did not offset the financial costs of culling. These results, combined with evaluation of alternative culling methods, suggest that badger culling is unlikely to contribute effectively to the control of cattle TB in Britain.

Repeated inhalation of crack-cocaine affects spermatogenesis in young and adult mice

To investigate the effects of repeated crack-cocaine inhalation on spermatogenesis of pubertal and mature Balb/c mice, ten young (Y-ex) and ten adult (A(ex)) Balb/c mice were exposed to the smoke from 5 g of crack with 57.7% of pure cocaine in an inhalation chamber, 5 days/week for 2 months. The young (Y-c) and adult (A(c)) control animals (n = 10) were kept in a specially built and controlled animal house facility. The morphologic analysis of both testes of all animals included the analysis of quantitative and qualitative histologic parameters to assess the effect of crack-cocaine on spermatogenesis and Leydig cells. Apoptosis was determined by immunolabeling with caspase-3 antibodies. Compared to the Y-c animals, Y-ex animals showed a significant reduction in the number of stage VII tubules per testis (p = 0.02), Sertoli cells (p < 0.001) and elongated spermatids (p = 0.001). Comparisons between the Y-ex and A(ex) groups identified a significant reduction in the number of Sertoli cells (p < 0.001) and round spermatids (p < 0.001) in the Y-ex group and a significant increase in apoptotic Leydig cells (p = 0.04) in the A(ex) group. The experimental results indicate that crack-cocaine smoke inhalation induced spermatogenesis disruption in chronically exposed mice, particularly in pubertal mice.

Effects of Repeated Stress on Distal Airway Inflammation, Remodeling and Mechanics in an Animal Model of Chronic Airway Inflammation

Background/Aims: Epidemiological studies suggest that stress has an impact on asthmatic exacerbations. We evaluated if repeated stress, induced by forced swimming, modulates lung mechanics, distal airway inflammation and extracellular matrix remodeling in guinea pigs with chronic allergic inflammation. Methods: Guinea pigs were submitted to 7 ovalbumin or saline aerosols (1-5 mg/ml during 4 weeks; OVA and SAL groups). Twenty-four hours after the 4th inhalation, guinea pigs were submitted to the stress protocol 5 times a week during 2 weeks (SAL-S and OVA-S groups). Seventy-two hours after the 7th inhalation, guinea pigs were anesthetized and mechanically ventilated. Resistance and elastance of the respiratory system were obtained at baseline and after ovalbumin challenge. Lungs were removed, and inflammatory and extracellular matrix remodeling of distal airways was assessed by morphometry. Adrenals were removed and weighed. Results: The relative adrenal weight was greater in stressed guinea pigs compared to non-stressed animals (p < 0.001). Repeated stress increased the percent elastance of the respiratory system after antigen challenge and eosinophils and lymphocytes in the OVA-S compared to the OVA group (p < 0.001, p = 0.003 and p < 0.001). Neither collagen nor elastic fiber contents were modified by stress in sensitized animals. Conclusions: In this animal model, repeated stress amplified bronchoconstriction and inflammatory response in distal airways without interfering with extracellular matrix remodeling. Copyright (C) 2011 S. Karger AG, Basel

Mechanical Performance of Cylindrical and Dual-Core Pedicle Screws After Repeated Insertion

Study Design. Ex vivo study of the mechanical performance of cylindrical and dual-core pedicle screws after insertion, removal, and reinsertion in the same hole. Objective. To evaluate the effect of repeated use of same screw hole on the insertion torque and the retentive strength of the cylindrical and dual-core screws. Summary of Background Data. Insertion and removal of pedicle screws is sometimes necessary during surgical procedure to assess the integrity of the pilot-hole wall. However, this maneuver may compromise the implant-holding capacity. Methods. Sixty thoracolombar vertebrae (T13-L5), harvested from 10 healthy calves, were used to insert 2 different designs of pedicle screws: cylindrical (5.0-mm outer diameter) and dual-core screws (5.2-mm outer diameter). Three experimental groups were created on the basis of the number of insertions of the screws and 2 subgroups were established according to the core pedicle screw design (dual-core and cylindrical). The insertion torque was measured during initial insertion, second insertion, and third insertion. Pullout screw tests were performed using a universal testing machine to evaluate the pullout strength after initial insertion, second insertion, and third insertion. Results. Significant reductions of 38% in mean insertion torque and 30% in mean pullout strength of dual-core screw were observed between the initial insertion and the third insertion. The cylindrical screw observed significant reductions of 52.5% in mean insertion torque and 42.3% in mean pullout strength between the initial insertion and the third insertion. A reduction of mean insertion torque and pullout strength between the first insertion and the second insertion but without significance was also observed for both types of screws. Conclusion. Insertions and reinsertion of either cylindrical or dual-core pedicle screws have compromised insertion torque and pullout strength of the implants as measured by mechanical tests.

Citrus aurantium L. essential oil exhibits anxiolytic-like activity mediated by 5-HT1A-receptors and reduces cholesterol after repeated oral treatment

Abstract Background The current treatments for anxiety disorders and depression have multiple adverse effects in addition to a delayed onset of action, which has prompted efforts to find new substances with potential activity in these disorders. Citrus aurantium was chosen based on ethnopharmacological data because traditional medicine refers to the Citrus genus as useful in diminishing the symptoms of anxiety or insomnia, and C. aurantium has more recently been proposed as an adjuvant for antidepressants. In the present work, we investigated the biological activity underlying the anxiolytic and antidepressant effects of C. aurantium essential oil (EO), the putative mechanism of the anxiolytic-like effect, and the neurochemical changes in specific brain structures of mice after acute treatment. We also monitored the mice for possible signs of toxicity after a 14-day treatment. Methods The anxiolytic-like activity of the EO was investigated in a light/dark box, and the antidepressant activity was investigated in a forced swim test. Flumazenil, a competitive antagonist of benzodiazepine binding, and the selective 5-HT1A receptor antagonist WAY100635 were used in the experimental procedures to determine the mechanism of action of the EO. To exclude false positive results due to motor impairment, the mice were submitted to the rotarod test. Results The data suggest that the anxiolytic-like activity observed in the light/dark box procedure after acute (5 mg/kg) or 14-day repeated (1 mg/kg/day) dosing was mediated by the serotonergic system (5-HT1A receptors). Acute treatment with the EO showed no activity in the forced swim test, which is sensitive to antidepressants. A neurochemical evaluation showed no alterations in neurotransmitter levels in the cortex, the striatum, the pons, and the hypothalamus. Furthermore, no locomotor impairment or signs of toxicity or biochemical changes, except a reduction in cholesterol levels, were observed after treatment with the EO. Conclusion This work contributes to a better understanding of the biological activity of C. aurantium EO by characterizing the mechanism of action underlying its anxiolytic-like activity.

Storage of bovine reproductive tissues and RNA extracts on ice for 24h or repeated freeze–thaw cycles do not affect RNA integrity

The aims of this study were to test (i) the effect of time of tissue and RNA extracts storage on ice and (ii) the effect of repeated freeze–thaw cycles on RNA integrity and gene expression of bovine reproductive tissues. Fragments of endometrium (ENDO), corpus luteum (CL) and ampulla (AMP) were subdivided and incubated for 0, 1, 3, 6, 12 or 24 h on ice. RNA extracts were incubated on ice for 0, 3, 12 or 24 h, or exposed to 1, 2, 4 or 6 freeze–thaw cycles. RNA integrity number (RIN) was estimated. Expression of progesterone receptor (PGR) and cyclophilin genes from RNA extracts stored on ice for 0 or 24 h, and 1 or 6 freeze–thaw cycles was measured by qPCR. Tissue and RNA extract incubation on ice, and repeated freeze–thaw cycles did not affect RIN values of RNA from ENDO, CL or AMP. Storage on ice or exposure to freeze–thaw cycles did not affect Cq values for PGR or cyclophilin genes. In conclusion, neither generalized RNA degradation nor specific RNA degradation was affected by storage of tissue or RNA extracts on ice for up to 24 h, or by up to 6 freeze–thaw cycles of RNA extracts obtained from bovine ENDO, CL and AMP.

Test-retest reliability of the nociceptive withdrawal reflex and electrical pain thresholds after single and repeated stimulation in patients with chronic low back pain

Recent studies have shown that the nociceptive withdrawal reflex threshold (NWR-T) and the electrical pain threshold (EP-T) are reliable measures in pain-free populations. However, it is necessary to investigate the reliability of these measures in patients with chronic pain in order to translate these techniques from laboratory to clinic. The aims of this study were to determine the test-retest reliability of the NWR-T and EP-T after single and repeated (temporal summation) electrical stimulation in a group of patients with chronic low back pain, and to investigate the association between the NWR-T and the EP-T. To this end, 25 patients with chronic pain participated in three identical sessions, separated by 1 week in average, in which the NWR-T and the EP-T to single and repeated stimulation were measured. Test-retest reliability was assessed using intra-class correlation coefficient (ICC), coefficient of variation (CV), and Bland-Altman analysis. The association between the thresholds was assessed using the coefficient of determination (r (2)). The results showed good-to-excellent reliability for both NWR-T and EP-T in all cases, with average ICC values ranging 0.76-0.90 and average CV values ranging 12.0-17.7%. The association between thresholds was better after repeated stimulation than after single stimulation, with average r (2) values of 0.83 and 0.56, respectively. In conclusion, the NWR-T and the EP-T are reliable assessment tools for assessing the sensitivity of spinal nociceptive pathways in patients with chronic pain.