Effects of synchronization of donor cell cycle on embryonic development and DNA synthesis in porcine nuclear transfer embryos

The relationship between donor cell cycle and the developmental ability of somatic cell nuclear transfer (SCNT) embryos has not fully been elucidated. Donor cells that are usually prepared by serum starvation or confluent-cell culture for SCNT represent a heterogeneous population that includes mainly G0 phase cells, other cells in different phases of the cell cycle and apoptotic cells. In this study, we compared the developmental ability of porcine SCNT embryos reconstructed from G0 phase cells (G0-SCNT embryos) and strictly synchronized-G1 phase cells (G1-SCNT embryos), and examined the developmental rates and timing of first DNA synthesis. The G0 phase cells were synchronized by confluent culture, and the G1 phase cells were prepared from actively dividing M phase cells. The G1-SCNT embryos showed a significantly higher (P<0.05) developmental rate to the blastocyst stage per cleaved embryo (59%) than the G0-SCNT embryos (43%). Moreover, initiation of first DNA synthesis and cleavage occurred significantly earlier in the G1-SCNT embryos than in the G0-SCNT embryos. Delay of initiation of first DNA synthesis in the SCNT embryos by aphidicolin resulted in decreased developmental rates to the blastocyst stage without any effect on cleavage rates. Our data demonstrates that synchronized-G1 phase cells can be used as donor cells for SCNT embryos and that earlier initiation of first DNA synthesis may be important for subsequent development of SCNT embryos. The SCNT system using G1-synchronized cells, in terms of their highly uniform and viable cell states, can be useful for studying the reprogramming processes and embryonic development of SCNT embryos.     

A new method for counting of quail leukocytes by flow cytometry

An automatic counting method was developed for fish blood cells using a fluorescent dye, 3, 3-dihexyloxacarbocyanine (DiOC (6)(3)), that selectively stain lipid bilayers in living cells. In the present study, the DiOC(6)(3) method was applied to quail (Cotumix cotumix japonica) blood cells. After quail blood cells were stained with DiOC(6)(3), absolute counts and relative proportions of erythrocytes, granulocytes, monocytes, and lymphocytes plus thrombocytes in whole blood were obtained by means of flow cytometry (FC). The number of each cell types by the FC was in good agreement with those counted microscopically. This method will offer new possibilities for routine blood cell counting for avian medicine.     

Effect of seasonal conditions on oestrus occurrence and postpartum period in Anatolian water buffaloes

Oestrus and calving records of Anatolian buffaloes were analyzed to investigate influence of climatic conditions on oestrus occurrence and postpartum period. Oestrus records showed a seasonal pattern concentrated between July and September which is the warmest period of the year. Likewise, more than half (52.7%) of the total calvings occurred between May and August. Month of calving significantly influenced the length of calving to first oestrus interval and open period (P < 0.01). The average length of open period was significantly shorter in autumn and summer calvers compared to those calved in spring and winter (P < 0.05) during long days. The humidity rate was negatively correlated with both calving to first oestrus interval and open period (P < 0.05). In conclusion, exposure to high environmental temperatures around 20 degrees C exerted no suppressive influence on ovarian activity in Anatolian buffaloes. Although humidity rate (P < 0.05) and ambient temperature (P < 0.01) was found to be correlated with the duration of postpartum period, day length might be the main factor regulating reproductive biorhythm and postpartum events in Anatolian buffalo cows.     

Diazepam potentiates the positive inotropic effects of histamine and forskolin in guinea-pig papillary muscles

There have been diverse reports on the effects of diazepam on cardiac contractility. The purpose of this study was to examine whether diazepam modifies the inotropic response elicited by histamine on an isolated guinea-pig papillary muscle. The responses of electrically driven papillary muscle to histamine and cyclic AMP-related inotropic agents were recorded in the absence and in the presence of diazepam. Histamine and forskolin, which directly stimulate adenylate cyclase, significantly increased the contractile force in the papillary muscle in a concentration-dependent manner. A histaminergic H₂-receptor antagonist, cimetidine, but not a H₁-receptor antagonist, diphenhydramine, at 10 μ M produced a rightward shift in the concentration-response curve for histamine. Diazepam (10 μ M ) shifted the concentration-response curve for histamine and forskolin to the left by 1.8 and 1.6 times, respectively. Neither a central type (fulmazenil) nor a peripheral type (PK11195) of benzodiazepine receptor antagonist modified the effect of diazepam on the histaminergic-evoked contraction. Phosphodiesterase blockade by 3-isobutyl-1-methylxanthine shifted the concentration-dependent curve for histamine to the left. A combination of 3-isobutyl-1-methylxanthine also produced a leftward shift of the curve. However, there was no significant difference between the 3-isobutyl-1-methylxanthine only group and the combination group. These results indicate that diazepam potentiates the positive inotropic effect produced by histamine, probably mediated via an increase in cyclic AMP levels induced by histamine.     

Effects of varying nitrogen doses on yield,yield components and artemisinin content of Artemisia annua L.

Artemisia annua L. is an aromatic-antibacterial herb that destroys malarial parasites, lowers fevers and checks bleeding, and of which the secondary compound of interest is artemisinin. The objective of the present study was to determine yield, yield components and artemisinin content of A. annua L. grown under four nitrogen applications (0, 40, 80 and 120 kg ha⁻¹) in the Çukurova region of Turkey in 2004 and 2005. Field trials were conducted at Çukurova University, Agricultural Faculty Field Crops Department. In the study, plant height, number of branches, fresh herbage yield, dry herbage yield, fresh leaf yield, dry leaf yield, essential oil content and artemisinin content (by high performance liquid chromatography, HPLC) were examined. By analysis of variance, nitrogen doses had no any statistical effect on the traits investigated except for artemisinin content. Artemisinin content of the dried leaves were significantly affected by nitrogen applications, which varied from 6.32 to 27.50 mg 100 g⁻¹. Contents were from 120 and 80 kg ha⁻¹ nitrogen for the years of 2004 and 2005, respectively.     

Phytochemical and antioxidant properties of anthocyanin-rich Morus nigra and Morus rubra fruits

In this study, phytochemical and antioxidant properties of anthocyanin-rich mulberry species of Morus nigra L. (black mulberry) and Morus rubra L. (red mulberry) fruits harvested from across Turkey were investigated. Fruit color, total phenolics (TP), total monomeric anthocyanin (TMA), titratable acidity (TA), and individual sugar and organic acid compositions were determined. Total antioxidant capacity (TAC) of fruits was assessed by both the trolox-equivalent antioxidant capacity (TEAC) and the ferric reducing antioxidant power (FRAP) assays. Black mulberry exhibited higher TP, TMA, TAC and TA when compared to red mulberry. The average TP contents of M. nigra and M. rubra were 2737 and 1603 μg gallic acid equivalent in g fresh weight basis (GAE/g fw), respectively. M. nigra had the richest amount of anthocyanin with an average of 571 μg cy-3-glu/g fw. Overall, TAC averaged 10.5 and 12.0 mmol TE/L by the TEAC and FRAP methods, respectively. We found that FRAP, TEAC, TP and TMA were significantly correlated (r = 0.64–0.99) with each other. Fructose (5.27 g/100 mL) and glucose (5.81 g/100 mL) were determined to be the major sugars in both mulberries. M. nigra displayed a higher TA (2.05 g/100 mL) than M. rubra (0.78 g/100 mL), with citric acid as the major acid.