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.     

Field Techniques in Zoo and Wildlife Conservation Work

Wildlife conservation fieldwork and field-like situations in zoo settings can be challenging to veterinarians and include restraining, sedating/anesthetizing, examining, and sampling wild animals. The lack of amenities and utilities such as running water or electricity, climatic conditions, biting insects, and even the general public can all be a hindrance when working with animals in this environment. Zoonotic diseases and local conflicts, as well as the work itself, can all be hazards. On the basis of our combined experience, with field examples set in Scotland, working practices and suggestions for fieldwork equipment and the processing of samples are outlined. Equipment, references for obtaining additional information, and considerations for capture and anesthesia in the field are also covered. The importance of good communications, within the animal care team, with stakeholders, and with the general public, is emphasized. Potential health and environmental hazards are discussed, with recommendations on protective clothing and resources to identify these hazards.     

Fish mortality due to acute ammonia exposure

Abstract. Fish mortality in the River Umia, near Villagarcia de Arosa, Pontevedra, Spain, which occurred after a discharge from a food sewage works, is studied. Physico-chemical parameters of water samples from sewage and from the River Umia were studied and concentrations of ammonia sufficiently high to cause toxic effects in fish (702 mg/1 and 302 mg/1 as total ammonia) were found. The toxicity of two concentrations of ammonia was investigated on goldfish, Carassius auratus (L.), using water quality values the same as found in the River Umia, to obtain the same concentrations of un-ionized ammonia (2·13 mg/1 and 0·91 mg/1). Fish exposed to high concentrations died within 24 h and three of six, which were exposed to the lowest concentration died within 96h. All fish at first showed symptoms of hyper excitability and hyperventilation, and at times of rest showed a decrease in respiration. Post-mortem findings generally included gill congestion and haemorrhage.     

Q fever in pregnant goats: humoral and cellular immune responses

Q fever is a zoonosis caused by the intracellular bacterium Coxiella burnetii. Both humoral and cellular immunity are important in the host defence against intracellular bacteria. Little is known about the immune response to C. burnetii infections in domestic ruminants even though these species are the major source of Q fever in humans. To investigate the goat’s immune response we inoculated groups of pregnant goats via inhalation with a Dutch outbreak isolate of C. burnetii. All animals were successfully infected. Phase 1 and Phase 2 IgM- and IgG-specific antibodies were measured. Cellular immune responses were investigated by interferon-gamma, enzyme-linked immunosorbent spot test (IFN-γ Elispot), lymphocyte proliferation test (LPT) and systemic cytokines. After two weeks post inoculation (wpi), a strong anti-C. burnetii Phase 2 IgM and IgG antibody response was observed while the increase in IgM anti-Phase 1 antibodies was less pronounced. IgG anti-Phase 1 antibodies started to rise at 6 wpi. Cellular immune responses were observed after parturition. Our results demonstrated humoral and cellular immune responses to C. burnetii infection in pregnant goats. Cell-mediated immune responses did not differ enough to distinguish between Coxiella-infected and non-infected pregnant animals, whereas a strong-phase specific antibody response is detected after 2 wpi. This humoral immune response may be useful in the early detection of C. burnetii-infected pregnant goats.     

Influence of the FecXR Allele in Heterozygous Ewes on Follicular Population and Outcomes of IVP and ET using LOPU‐Derived Oocytes

Ewes heterozygous for the FecX^R allele (R+) in the bone morphogenetic protein 15 (BMP15) gene display increased ovulation rate and prolificacy. Besides this phenotypic advantage, the influence of the FecX^R allele on follicle number and size, oocyte competence and in vitro production (IVP) remains undefined. With these aims, 8 R+ and 8 wild‐type (++) ewes were subjected to 2 laparoscopic ovum pick‐up (LOPU) trials (four sessions per trial; two with and two without FSH) and subsequent IVP and fresh embryo transfer. All follicles >3 mm were punctured (n = 1673). Genotype did not significantly affect the number of punctured follicles per ewe and session (10.4 and 10.2 in R+ and ++ untreated ewes, 17.4 and 14.3 in R+ and ++ FSH‐treated ewes, respectively), but follicular diameter of R+ ewes was significantly reduced compared with ++ ewes (?0.2 mm in untreated and ?0.8 mm in FSH‐treated ewes; p < 0.01). R+ ewes showed higher recovery rate and increased numbers of total and suitable cumulus–oocyte complexes for in vitro maturation (IVM). Similar rates of day 8 blastocysts were observed in R+ (36.1%, 147/407) and ++ (32.6%, 100/307) ewes, but the final output of day 8 blastocysts per ewe and session was higher in R+ ewes (+0.75; p < 0.005), without differences in survival rate at birth of the transferred embryos (40.4%, 21/52 vs 36.4%, 16/44, respectively). In conclusion, a higher number of oocytes proven to be competent for in vitro development and embryo survival after transfer are recovered from R+ ewes, despite the lower mean size of their follicles at puncture.