Comparison of three methods including temperature, H2O2/ascorbic acid/sonication, and nitrous acid treatments for overcoming the inhibitory effect of heparin on DNA amplification in realtime-PCR.

Heparin molecules have an inhibitory effect on DNA amplification by binding to the majority of DNA-interacting proteins. Different physical, chemical, and enzymatic methods have been used to degrade and depolymerize heparins in biomedical investigations. In this study, we aimed to evaluate some heparin degradation methods to eliminate the inhibitory effect of heparin on DNA amplification. Here, we report highly efficient, simple, and convenient methods to eliminate the heparin inhibitory effect on DNA amplification by treatments including temperature, nitrous acid, and H2O2/ascorbic acid/sonication. Further, treatment conditions including temperature degree and duration of treatments, the concentration of ascorbic acid, and intensity of sonication were reviewed. Target DNAs were extracted using the phenol-chloroform method. DNA concentrations and purity were analyzed before and after each treatment by Nanodrop spectrophotometry. DNA amplifications were attempted using a commercially available realtime-PCR mastermix. We found that the inhibitory behavior of heparin was well eliminated after the 85 °C/2 h, 65 °C/2 h, nitrous acid (pH = 3), and H2O2/ascorbic acid/sonication treatments, respectively. The further analyses indicated that the application of nitrous acid in pH = 1.5 and H2O2/ascorbic acid/sonication in higher ascorbic acid concentrations and sonication intensities lead to failure in DNA amplification due to the degradation of target sequences. From our experience, simple heat treatments or at the next level using nitrous acid and H2O2/ascorbic acid/sonication have enabled the detection and quantification of virus infection in heparin blood samples. These approaches may enable researchers to utilize blood taken in heparin tubes for genome amplification and diagnostic purposes.

Data for zonisamide effect on length and weight of 14 day-old rat pups and abortion rate and day in refractory epileptic pregnant rats.

Length and weight of 14 day-old rat pups and also abortion rate and day on refractory epileptic pregnant rats after treatment with zonisamide (ZNS) are presented. Lamotrigine-resistant chemical kindling procedure was used for inducing of refractory epilepsy. For further interpretation follow the research article: Effect of zonisamide on refractory epilepsy during pregnancy in lamotrigine resistant kindled rats (Sani et al., 2017) [1].

Effects of different regimens of PGF2α treatment during postpartum on reproductive performance in dairy cows / Efeito de diferentes regimes de tratamento pós-parto com PGF2α sobre o desempenho reprodutivo de vacas leiteiras

The aim of this study was to evaluate the effects of early postpartum PGF2α treatment on reproductive performance in dairy cows synchronized with targeted breeding and natural mating after voluntary waiting period. In this experiment ,120 cows were assigned to three groups irrespective of presence or absence of luteal tissue. Cows in PG- 14 group were treated with PGF2α from day 14 postpartum, cows in PG-28 group were treated with PGF2α from day 28 postpartum and cows in PG-42 group were not treated with PGF2α until the end of voluntary waiting period (d 42). After day 42 postpartum, cows in three groups were treated with PGF2α within 14-day intervals until natural mating after voluntary waiting period. Recorded reproductive parameters included days to first heat, days to first mating, days open, service per conception, conception rate, percentage of repeat breeder animals and pregnancy loss. Early PGF2α treatment from day 14 postpartum significantly decreased days to first estrus (34.9 ± 0.74, P < 0.003), days to first mating (62.35 ± 1.53, P < 0.04), days open (117.23 ± 3.1, P < 0.001) and service per conception (1.9 ± 0.09, P < 0.02); and PG-14 group presented increased conception rate (52.5%, P < 0.05). The proportion of repeat breeder syndrome tended to be affected by treatment with PGF2α from day 14 postpartum. In conclusion, treatment of cows with PGF2α from day 14 postpartum improved reproductive performance.(AU)

O objetivo do presente trabalho foi avaliar os efeitos do tratamento pós-parto precoce com PGF2α sobre o desempenho reprodutivo de vacas leiteiras sincronizadas para reprodução controlada por monta natural após o período de espera voluntário. Neste experimento, 120 vacas foram distribuídas em três grupos independentes da presença ou ausência de corpo lúteo. Vacas no grupo PG-14 foram tratadas com PGF2α a partir do 14o dia pósparto, vacas do grupo PG-28 foram tratadas com PGF2α a partir do 28o dia pós-parto e as vacas do grupo PG-42 não foram tratadas com PGF2α até o final do período de espera voluntário (d42). Após o 42o dia pós-parto as vacas dos três grupos foram tratadas com PGF2α com intervalos de 14 dias até a monta natural após o período de espera voluntário. Os registros dos parâmetros reprodutivos incluíram: dias para o primeiro estro, dias para a primeira cobertura, dias em aberto, serviços por concepção, taxa de concepção, percentagem de animais repetidores de cios e as perdas de gestações. O tratamento precoce com PGF2α, a partir do 14o dia pós-parto reduziu significativamente os dias para o primeiro estro (34,9 ± 0,74, P < 0,003), dias para a primeira cobertura (62,35 ± 1,53, P < 0,04), dias em aberto (117,23 ± 3,1, P< 0,02); e o grupo PG-14 apresentou um acréscimo na taxa de concepção (52,5%, P < 0,05). A proporção da síndrome de vacas repetidoras de cios tendeu a ser afetada pelo tratamento com PGF2α a partir do 14o dia pós-parto. A conclusão obtida foi que o tratamento das vacas com PGF2α a partir do 14o dia pós-parto melhorou o desempenho reprodutivo dos animais.(AU)