Kinome profiling of Arabidopsis using arrays of kinase consensus substrates

Background  

Kinome profiling aims at the parallel analysis of kinase activities in a cell. Novel developed arrays containing consensus substrates for kinases are used to assess those kinase activities. The arrays described in this paper were already used to determine kinase activities in mammalian systems, but since substrates from many organisms are present we decided to test these arrays for the determination of kinase activities in the model plant species Arabidopsis thaliana.     

Some blood parameters of the Chinese grasscarp Ctenopharyngodon idella (Valenciennes)

The activity cycle of a captive group of Dendrohyrax arboreus comprising an adult male, an adult female and two juveniles showed them to be more active during the daylight hours than nocturnally. Activities of the siblings were closely synchronized, but showed no close relationship to the mother's cycle. The animals spent most of their time on the upper level of their enclosure. The data support the view that D. arboreus is arboreal and solitary in its natural habitat. The nocturnal habits of some wild D. arboreus populations may represent a shift in activity pattern in response to hunting pressure by man.     

Relative accuracy of the identification of ovarian structures in the cow by ultrasonography and palpation per rectum

Manual palpation or ultrasonographic examination of the cow's genital tract are currently used by veterinarians involved in reproductive management, but knowledge of the potential and the limitations of both methods is important to obtain an optimal accuracy in the diagnosis of physiological and pathological ovarian structures. This review presents the main features of manual and ultrasonographic characteristics of follicles, corpora lutea (with or without a cavity), follicular and lutenized cysts and the reliability of the two methods is compared. Manual diagnosis of follicles <10 mm is rather inaccurate, but ultrasound offers the possibility to diagnose follicles <5 mm and to measure their inner diameter. The predictive values of the presence or absence of a corpus luteum as determined by palpation are similar (78 vs. 75%). Manual or ultrasonographic diagnosis of the growing or regressing corpus luteum is rather difficult. The positive predictive value of a mature corpus luteum diagnosed by ultrasonography is lower (87%) than the negative predictive value (92%). Compared to manual palpation, ultrasonography permits a better estimation of the number and to determine more precisely the size of the mature corpus luteum. The positive values for follicular cysts diagnosed by palpation or by ultrasonography are 66 and 74% respectively, and for luteal cysts, the values are 66 and 85%, respectively.Manual palpation or ultrasonography are useful tools to diagnose ovarian structures in the cow. The accuracy of such methods can be enhanced by securing information on the reproductive history of the animal, palpation of uterine horns, vaginal examination or progesterone determinations.     

13,14‐Dihydro‐15‐Keto Prostaglandin F2α Release in Response to Oxytocin Challenge Early Post‐Partum in Anoestrous Nelore Cows Submitted to Temporary Calf Removal and Progesterone Priming

The study evaluated, in early post‐partum anoestrous Nelore cows, if the increase in plasma oestradiol (E2) concentrations in the pre‐ovulatory period and/or progesterone priming (P4 priming) preceding ovulation, induced by hormonal treatment, reduces the endogenous release of prostaglandin PGF₂αand prevents premature lysis of the corpus luteum (CL). Nelore cows were subjected to temporary calf removal for 48 h and divided into two groups: GPE/eCG group (n = 10) and GPG/eCG group (n = 10). Animals of the GPE/eCG group were treated with a GnRH agonist. Seven days later, they received 400 IU of eCG, immediately after PGF₂α treatment, and on day 0, 1.0 mg of oestradiol benzoate (EB). Cows of the GPG/eCG group were similarly treated as those of the GPE/eCG group, except that EB was replaced with a second dose of GnRH. All animals were challenged with oxytocin (OT) 9, 12, 15 and 18 days after EB or GnRH administration and blood samples were collected before and 30 min after OT. Irrespective of the treatments, a decline in P4 concentration on day 18 was observed for cows without P4 priming. However, animals exposed to P4 priming, treated with EB maintained high P4 concentrations (8.8 ± 1.2 ng/ml), whereas there was a decline in P4 on day 18 (2.1 ± 1.0 ng/ml) for cows that received GnRH to induce ovulation (p < 0.01). Production of 13,14‐dihydro‐15‐keto prostaglandin F₂α (PGFM) in response to OT increased between days 9 and 18 (p < 0.01), and this increase tended to be more evident in animals not exposed to P4 priming (p < 0.06). In conclusion, the increase in E2 during the pre‐ovulatory period was not effective in inhibiting PGFM release, which was lower in P4‐primed than in non‐primed animals. Treatment with EB promoted the maintenance of elevated P4 concentrations 18 days after ovulation in P4‐primed animals, indicating a possible beneficial effect of hormone protocols containing EB in animals with P4 priming.     

Development and spatial distribution of the free-living stages of Teladorsagia circumcincta and Trichostrongylus colubriformis on pasture: A pilot study

Abstract

AIMS: To measure the development of Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis eggs to third-stage infective larvae (L3) at different times of the year. Also, to measure the spatial distribution of L3 across herbage, soil and faeces, in order to assess whether spatial issues could be important in larval dynamics on pasture.

METHODS: Field plots were contaminated with sheep faeces containing approximately 20,000 eggs of each of T. circumcincta and T. colubriformis on five separate occasions, viz 01 December 1996 (summer), 18 March 1998 (autumn), 17 June 1998 (winter), 15 October 1998 (spring), and 23 July 1999 (winter). Replicate plots (n=10) were harvested at intervals for up to 12 months after deposition of faeces, and the number and distribution of L3 were measured. Larvae were sampled from faeces (where these remained), herbage, and three soil zones to a depth of 145 mm.

RESULTS: There were large differences between contamination dates in the percentage of eggs that developed to L3. For both species the highest percentage development was for eggs deposited in December (7.8% and 25.9% for T. circumcincta and T. colubriformis, respectively) and the lowest for June (0.4% and 0.03% T. circumcincta and T. colubriformis, respectively). Development in winter was often delayed, and this was always associated with a low yield of larvae, probably due to compounding mortalities associated with long periods of exposure to low temperatures.

The relative distribution of L3 present on herbage, in faeces or in the soil varied between sampling times. However, overall the most L3 were recovered from soil (74% and 66% for T.circumcincta and T. colubriformis, respectively, averaged over all samples), and the lowest recoveries were from the herbage.

CONCLUSIONS: Although the data are limited, the results indicated that the highest percentage of eggs developed to infective larvae in summer and only minimal development occurred in winter. The data do not support the view that substantial contamination of pastures with sheep parasites occurs over winter. Large numbers of larvae were recovered from soil, which indicates that, assuming they can subsequently migrate onto herbage, soil is a potentially important reservoir ofinfective larvae in New Zealand. Therefore, the spatial distribution of L3 on pasture may affect both the dynamics and transmission of parasite populations. Further work on both these issues is warranted.     

Detection of corpora lutea and follicles in cows: a comparison of transvaginal ultrasonography and rectal palpation

The ovaries of 59 pluriparous cows of unknown reproductive history were palpated, scanned and dissected on the day of slaughter to compare the accuracy of rectal palpation and transvaginal ultrasonography with a 5 MHz linear array for the detection of corpora lutea and follicles. The rectal palpation was first carried out to judge the presence of follicles of more than 5 mm diameter, and corpora lutea which were classified as young (days 1 to 4), mid-cycle (days 5 to 16) or old (days 17 to 21) according to morphological criteria. The cows were then examined for follicles and corpora lutea by ultrasonography and the corpora lutea were again classified directly as young, mid-cycle or old according to their appearance. The cows were then slaughtered, their ovaries dissected, and the follicles over 5 mm in diameter were counted and the corpora lutea were classified in the above mentioned age categories. For the detection of a mid-cycle corpus luteum the sensitivity and predictive value of rectal palpation were, respectively, 83.3 per cent and 73.2 per cent and for ultrasonography the sensitivity and predictive value were 80.6 per cent and 85.3 per cent, respectively. However, both techniques were inaccurate for the detection of young and old corpora lutea. For detecting follicles ultrasonography was a significantly better method than rectal palpation. Ultrasonography detected 95 per cent of follicles larger than 10 mm whereas rectal palpation detected only 71 per cent of these follicles. Both techniques failed with follicles 5 to 10 mm in diameter; only 21.5 per cent were detected by rectal palpation and 34.3 per cent by ultrasonography.