An insight into seed priming response of Crotalaria ochroleuca and Crotalaria spectabilis during storage.

The proper establishment of plants is essential for the efficient use of resources such as water and light. Besides, even after seed storage and sowing the uniform establishment of plants is essential for their success. Crotalaria ochroleuca and Crotalaria spectabilis are important medicinal plants with poor seed germination rate, occasionally. The effects of seed priming in both C. ochroleuca and C. spectabilis were evaluated in seed performance even after seeds storage for up 90-days. Experimental assays were performed in a randomized design with gibberellic acid (GA3, 100 ppm), polyethylene glycol (PEG 6000, -0.2 MPa) and PEG (-0.2 MPa) + GA3 (100 ppm) solutions during seed priming in four replicates. Seeds not submitted to priming treatments constituted control. Seeds physiological performance were evaluated immediately and even after 30, 60 and 90-days seed dry-storage. The data obtained in each experiment were submitted to variance analysis (ANOVA) adopting a confidence level of 95%. The effects of seed priming with PEG and GA3 during seed ageing were significant for germination variables of C. ochroleuca and C. spectabilis. During dry storage, seed viability of both species gradually decreased and the first symptoms were delayed seed germination, especially more evident for C. ochroleuca, even in primed or non-primed seeds. Afterwards, C. ochroleuca seeds previously GA3 primed had higher results of root protrusion (86%), hypocotyls elongation (76%) and complete seedlings (75%) than non-primed seeds (control). These findings shown a good potential of hormopriming to attenuate damage during the seed aging of C. ochroleuca.

Addition of chlorogenic acid and caffeine during the processing of cooled boar semen / Adição de ácido clorogênico e cafeína durante o processamento do sêmen suíno resfriado

A study was conducted to evaluate the effect of chlorogenic acid (ChA) added pre-cooling and its combination with caffeine added during warming on cooled-stored boar semen parameters. Ten ejaculates were diluted in commercial extender with or without 4.5mg/ml ChA and stored at 15°C. After 0, 24 and 72 hours of storage, aliquots of these doses were taken and incubated at 37°C in the presence or absence of 8.0mM caffeine. Semen quality was evaluated after 10 and 120 minutes of incubation. The ChA increased (P <0.01) the sperm motility, viability, acrosomal integrity and the percentage of spermatozoa with high mitochondrial activity (PMHA), however, decreased (P <0.01) the malondialdehyde (MDA) concentration. Caffeine increased (P<0.05) the sperm motility, viability, PMHA and the MDA concentration and reduced (P <0.05) the acrosome integrity. When associated (ChA+caffeine), there was an increase (P <0.05) in sperm motility and viability, PMHA and acrosome integrity. The addition of ChA to the dilution medium improves the quality of the swine inseminating doses. The addition of caffeine during re-warming is only recommended when the semen is stored for prolonged periods (72h), and the inseminating dose should be used immediately after its addition.(AU)

O objetivo deste estudo foi avaliar os efeitos da adição de ácido clorogênico (ChA) antes do resfriamento e sua combinação com cafeína adicionada durante o reaquecimento sobre a qualidade do sêmen suíno resfriado. Dez ejaculados foram diluídos em diluidor comercial com adição ou não de 4,5mg/mL de ChA e armazenados a 15°C. Após zero, 24 e 72 horas de armazenamento, 10mL foram retirados e incubados a 37°C na presença ou ausência de 8,0mM de cafeína. A qualidade seminal foi avaliada após 10 e 120 minutos de incubação. O ChA aumentou (P<0,01) a motilidade, a viabilidade, a integridade acrosomal e a porcentagem de espermatozoides com alta atividade mitocondrial (PMHA), entretanto diminuiu (P<0,01) a concentração de malondialdeído (MDA). A cafeína aumentou (P<0,05) a motilidade, a viabilidade, a PMHA e a concentração de MDA e reduziu a integridade acrossomal. Quando associados (ChA+cafeína), houve aumento (P<0,05) na motilidade, na PMHA, na viabilidade e na integridade acrossomal. Conclui-se que a adição de ChA ao meio de diluição melhora a qualidade das doses inseminantes de suínos. A adição de cafeína durante o reaquecimento só é recomendada ao sêmen adicionado de ChA quando esse for armazenado por períodos prolongados (72h), devendo a dose inseminante ser utilizada imediatamente após sua adição.(AU)

Incidence of the second parity syndrome in sows from a commercial farm / Incidência de síndrome do segundo parto em fêmeas suínas de uma granja comercial

O objetivo do presente estudo foi investigar a ocorrência da síndrome do segundo parto em uma granja comercial de suínos e apresentar alternativas para minimizar esse problema reprodutivo. Os dados foram obtidos de 363 fêmeas de genética comercial (DB-30) de primeiro e segundo partos, entre os anos de 2010 e 2011. Os animais pertenciam a uma granja comercial de ciclo completo com 1200 matrizes, cujos índices zootécnicos não permitiam detectar a presença da síndrome do segundo parto. O período de lactação foi de 24,6±3,3 dias. Foram analisados o número de nascidos totais e nascidos vivos, o peso da leitegada ao nascimento, o número de desmamados e o peso ao desmame do lote e também individualmente de cada marrã ao longo do ano. As médias e o desvio-padrão foram calculados, e os dados obtidos no primeiro e no segundo parto foram comparados pelo teste t pareado a 5%. Não houve diferença (P>0,05) no número de nascidos totais e no número de nascidos vivos entre o primeiro e o segundo parto. No entanto, constatou-se que 54% das fêmeas apresentaram igual ou menor número de nascidos no segundo parto, caracterizando a síndrome do segundo parto na maior parte dos animais. Nesse lote, o número de leitões nascidos a menos em relação ao primeiro ciclo reprodutivo foi de 3,6±2,9. Das 363 matrizes avaliadas, 153 (42%) apresentaram 16 ou mais leitões no primeiro parto. Destas, 92 (60%) tiveram menor número de leitões no segundo parto e 41 (27%) apresentaram maior número de leitões. Também se verificou maior incidência (50% ou mais) da síndrome do segundo parto nos meses de janeiro a março e de outubro a dezembro. Conclui-se que a síndrome do segundo parto é um problema que pode afetar 50% ou mais das matrizes, nem sempre detectada por meio dos índices zootécnicos da granja. Medidas como pesagem dos animais na primeira cobertura e logo após o desmame, além de programas de alimentação com dietas balanceadas, principalmente durante os meses mais quentes do ano, são ferramentas importantes para amenizar esse problema.(AU)

Intracellular calcium movements of boar spermatozoa during 'in vitro' capacitation and subsequent acrosome exocytosis follow a multiple-storage place, extracellular calcium-dependent model.

This work analysed intracellular calcium stores of boar spermatozoa subjected to 'in vitro' capacitation (IVC) and subsequent progesterone-induced acrosome exocytosis (IVAE). Intracellular calcium was analysed through two calcium markers with different physico-chemical properties, Fluo-3 and Rhod-5N. Indicative parameters of IVC and IVAE were also evaluated. Fluo-3 was located at both the midpiece and the whole head. Rhod-5N was present at the sperm head. This distribution did not change in any of the assayed conditions. Induction of IVC was concomitant with an increase in both head and midpiece Ca(2+) signals. Additionally, while IVC induction was concurrent with a significant (p < 0.05) increase in sperm membrane permeability, no significant changes were observed in O2 consumption and ATP levels. Incubation of boar spermatozoa in the absence of calcium showed a loss of both Ca(2+) labellings concomitantly with the sperm's inability to achieve IVC. The absence of extracellular calcium also induced a severe decrease in the percentage of spermatozoa exhibiting high mitochondrial membrane potential (hMMP). The IVAE was accompanied by a fast increase in both Ca(2+) signalling in control spermatozoa. These peaks were either not detected or much lessened in the absence of calcium. Remarkably, Fluo-3 marking at the midpiece increased after progesterone addition to sperm cells incubated in a medium without Ca(2+) . The simultaneous addition of progesterone with the calcium chelant EGTA inhibited IVAE, and this was accompanied by a significant (p < 0.05) decrease in the intensity of progesterone Ca(2+) -induced peak, O2 consumption and ATP levels. Our results suggest that boar spermatozoa present different calcium deposits with a dynamic equilibrium among them and with the extracellular environment. Additionally, the modulation role of the intracellular calcium in spermatozoa function seems to rely on its precise localization in boar spermatozoa.

Cryotolerance of stallion spermatozoa is related to ROS production and mitochondrial membrane potential rather than to the integrity of sperm nucleus.

Although cryopreservation of stallion spermatozoa allows long-term preservation of spermatozoa from particular stallions and facilitates international trade, it is understood to inflict damages on sperm cells that may finally reduce their fertilizing ability. In addition, individual differences are known to exist in the sperm ability to withstand freeze-thawing protocols. To date, these differences have mainly been reported on the basis of sperm motility and membrane integrity. For this reason, the present work sought to determine differences between good (good freezability ejaculates: GFE) and poor (poor freezability ejaculates: PFE) freezability stallion ejaculates in other sperm parameters, including peroxide and superoxide levels, potential of mitochondrial membrane and nuclear integrity. With this purpose, a total of 24 stallion ejaculates were cryopreserved and classified into two groups (GFE vs. PFE), depending on their sperm membrane integrity and motility after freeze-thawing. From the total of 24 ejaculates, 13 were classified as GFE and the other 11 were classified as PFE. Apart from differences in sperm membrane permeability and lipid disorder after freeze-thawing, GFE presented significantly (p < 0.05) higher percentages of viable spermatozoa with high content of peroxides and of superoxides than PFE. In contrast, and despite cryopreservation of stallion spermatozoa increasing DNA fragmentation and disrupting disulphide bonds in sperm head proteins, no significant differences between GFE and PFE were seen. We can thus conclude that good and poor freezability stallion ejaculates differ in their reactive oxygen species levels after cryopreservation, but not in the damage extent on sperm nucleus.

Supplementing cryopreservation media with reduced glutathione increases fertility and prolificacy of sows inseminated with frozen-thawed boar semen.

The main aim of this work was to evaluate how supplementing freezing media with reduced glutathione (GSH) affected the 'in vivo' fertilizing ability of boar semen subjected to cryopreservation procedures. With this purpose, 12 ejaculates coming from 12 boars were cryopreserved in the presence or absence of 2 mm GSH, whereas the same number of extended ejaculates coming from the same boars was used as negative/farm controls. Eight different sperm parameters (levels of free-cysteine residues in sperm nucleoproteins, DNA fragmentation, sperm viability, acrosome-membrane integrity, intracellular peroxide and superoxide levels, and total and progressive sperm motility) were evaluated before freezing and after 30 and 240 min of thawing. In addition, a total of 180 multiparous sows were used in the field fertility trials, the females being randomly divided into three groups and inseminated with extended, frozen-thawed control or frozen-thawed semen supplemented with 2 mm GSH. The presence of GSH in the freezing media significantly (p < 0.05) increased farrowing rates and the number of total born piglets and alive born piglets, and partially counteracted the cryopreservation-induced damages inflicted on frozen-thawed spermatozoa. We can thus conclude that supplementing freezing media with 2 mm GSH greatly improves boar sperm cryopreservation technology, as it significantly improves the fertilizing ability of frozen-thawed spermatozoa.

Effect of the addition of IGF-I and vitamin E to stored boar semen.

The objective of this study was to evaluate the addition of IGF-I to pig insemination doses stored at 15°C, in conjunction with the addition of different amounts of vitamin E (α-tocopherol). Semen samples (n = 12) from four boars were treated by the addition of different concentrations of vitamin E, ranging up to 400 µg/ml. Immediately after processing and after the doses had been stored at 15°C for 24 or 72 h, samples were warmed at 37°C and 30 ng/ml of IGF-I was added. The assessments were made after 10 and 120 min of IGF-I addition. There was a minor effect of the vitamin E added before cooling and IGF-I added after storage on sperm quality. The addition of 400 µg/ml of vitamin E to diluted semen reduced (P < 0.01) the malondialdehyde (MDA) production in boar semen stored at 15°C for 72 h, regardless of the addition of IGF-I as additive during a 120 min incubation period at 37°C. In these conditions, IGF-I also reduced (P < 0.05) the MDA production in semen samples without addition of vitamin E. IGF-I in the presence of vitamin E reduced (P = 0.03) the glucose intake in freshly diluted boar semen samples before cooling. It was concluded that the addition of 400 µg/ml of vitamin E reduces the MDA production in boar semen stored at 15°C for 72 h, regardless of the presence of IGF-I additive. The addition of IGF-I in doses stored for 72 h with vitamin E ensures higher sperm motility after 120 min of incubation at 37°C.

Addition of IGF-I to storage-cooled boar semen and its effect on sperm quality.

OBJECTIVE: To evaluate in vitro IGF-I treatment during warming of storage-cooled boar semen and its effect on seminal quality parameters and metabolism in spermatic cells. DESIGN: Semen samples (n=7) warmed after stored at 15°C for 24 or 72h were divided into four equal parts. Different IGF-I concentrations (0, 50, 100 and 150ng/mL) were added to the semen samples. The samples were incubated at 37°C, and assessments were made after 0 and 120min of incubation. RESULTS: For semen samples that were stored for 24h, the addition of IGF-I had no effect (p>0.05) on the total motility and intensity of movements by spermatic cells, osmotic resistance, live:dead cell ratio or total spermatic abnormalities. However, incubation with 150ng/mL IGF-I did decrease glutathione peroxidase activity (p<0.05) and reduce lipid peroxidation after 120min of incubation. For semen samples stored for 72h and incubated with IGF-I for 120min, there was a linear relationship between the IGF-I concentration and the live:dead ratio (p<0.05). There was a quadratic relationship between the IGF-I concentration and both the osmotic resistance (peak results at IGF-I=62.4ng/mL) and glutathione peroxidase activity (peak results at IGF-I=77.8ng/mL). There was no effect on lipid peroxidation (p>0.05) after 120min of incubation. Addition of IGF-I also decreased fructose utilization by spermatic cells regardless of semen storage time (p<0.05). CONCLUSION: This study suggests that IGF-I may be beneficial to semen stored for longer periods of time. Adding 150ng/mL IGF-I improved the quality of semen stored for 24h, and adding 78ng/mL IGF-I improved the quality of semen stored for 72h.

Decrease in oxidative stress and histological changes induced by physical exercise calibrated in rats with osteoarthritis induced by monosodium iodoacetate.

OBJECTIVE: The purpose of this study was to evaluate the effects of impact exercise on the joint cartilage of rats with osteoarthritis (OA) induced by monosodium iodoacetate (MIA). METHODS: Eighteen male rats were divided into three groups of six animals each: control, OA, and OA plus exercise (OAE). The OAE group trained on a treadmill for 8 weeks. Afterward, the right joints of the animals were washed with saline solution and joint lavage was used for biochemical analyses of myeloperoxidase (MPO) and enzyme superoxide dismutase (SOD) activities and total thiol content. The same limb provided samples of the articular capsule for analyses of MPO activity and total thiol content. The left joint was used for histological analysis. RESULTS: Our results indicate that MPO activity was increased in both OA groups in the lavage as well as the articular capsule, regardless of exercise status. SOD activity was increased in animals with OA, especially in the animals that had run on the treadmill. On the other hand, thiol content in the articular capsule and joint lavage decreased in the OA group, while the OAE group had values similar to those of the control group. The histological data indicate that animals that were submitted to running exercise showed a higher preservation rate of proteoglycan content in the superficial and intermediate areas of the joint cartilage. CONCLUSION: Our results show that physical training contributes to the preservation of joint cartilage in animals with OA and to increase the defense mechanism against oxidative stress.

The effect of the Rochagan on radiolabeling with (99m)Tc.

Radionuclides are used in nuclear medicine by variety of diagnostic procedures. The labeling of red blood cells (RBC) with (99m)Tc is a current method applied in clinical nuclear medicine. Drugs can alter this labeling and modify the disposition of the radiopharmaceuticals. The influence of Rochagan on the labeling of blood constituents with (99m)Tc was reported. Samples of blood were incubated with different concentrations of Rochagan (0%; 6.25%; 12.5%; 25%; 50%; 100%). Stannous chloride and (99m)Tc (3.7MBq/mL) were added. Plasma (P) and (RBC) were isolated and precipitated with thricloroacetic acid 5%. The insoluble (IF) and soluble fractions (SF) were separated. The %ATI in RBC, IF-P and IF-RBC were calculated. The %ATI on RBC decreased significantly (p<0.05) from control to all concentrations of Rochagan, respectively: 90.15 + or - 0.14(control) to 70.80 + or - 4.21; to 64.36 + or - 0.33; to 57.30 + or - 1.56; to 50.28 + or - 2.71; to 42.41 + or - 2.24; on IF-RBC, respectively: 84.70 + or - 0.87(control) to 67.16 + or - 4.38; to 63.63 + or - 2.92; to 59.02 + or - 3.17; to 43.75 + or - 1.00; to 24.15 + or - 0.94 and also on IF-P, respectively: 83.46 + or - 1.09(control) to 50.90 + or - 3.36; to 35.46 + or - 4.13; to 35.78 + or - 2.31; to 28.74 + or - 3.09; to 19.66 + or - 1.34. The analyses were performed by T-Student and Mann Whitney tests, p<0.05. This effect was probably due to products present in Rochagan that may complex with ions or have a direct/indirect effect on intracellular stannous ion concentration.

A review of natural products with antileishmanial activity.

Infections caused by protozoa of the genus Leishmania are a major worldwide health problem, with high endemicity in developing countries. The incidence of the disease has increased since the emergence of AIDS. In the absence of a vaccine, there is an urgent need for effective drugs to replace/supplement those in current use. The plant kingdom is undoubtedly valuable as a source of new medicinal agents. The present work constitutes a review of the literature on plant extracts and chemically defined molecules of natural origin showing antileishmanial activity. The review refers to 101 plants, their families, and geographical distribution, the parts utilized, the type of extract and the organism tested. It also includes 288 compounds isolated from higher plants and microorganisms, classified into appropriate chemical groups. Some aspects of recent antileishmanial-activity-directed research on natural products are discussed.

Propsonina em criancas desnutridas com septicemia. / Propsonine in undernourished children with septicemia

Os autores utilizaram no IPPMG da UFRJ um produto novo, a propsonina, em 32 criancas desnutridas apresentando quadro de septicemia ou infeccao, como coadjuvante de terapeutica antimicrobiana convencional quando esta nao atuou eficazmente. Comparando este grupo com 32 criancas de um grupo-controle, os autores viram que o periodo de internacao foi mais curto e a taxa de mortalidade baixou sensivelmente.Nao houve colateral em um total de duzentas e oitenta e duas doses administradas.Fundamentados neste trabalho inicial os autores, puderam tirar algumas conclusoes interessantes como a importancia do aleitamento materno, o grau de desnutricao e a faixa etaria, contribuindo para maior frequencia de septicemia em criancas. Terminam os autores achando que deveriam aumentar o numero de casos para futura analise estatistica e conlusoes definitivas