Cellular and extracellular vesicular origins of miRNAs within the bovine ovarian follicle

The ovarian follicle components must provide an ideal environment to ensure the success of reproductive processes, and communication between follicular cells is crucial to support proper oocyte growth. Recently, it has been demonstrated that the presence of extracellular vesicles (EVs) carrying microRNAs (miRNAs) in follicular fluid represents an important autocrine and paracrine communication mechanism inside the ovarian follicle. In this study, we tested the hypothesis that the miRNA content of EVs isolated from ovarian follicular (granulosa and cumulus–oocyte complexes) cell‐conditioned culture media is dependent upon cell type. We initially screened bovine granulosa cells (GCs) and cumulus–oocyte complexes (COCs), as well as their derived EVs for 348 miRNAs using real‐time PCR, and detected 326 miRNAs in GCs and COCs cells and 62 miRNAs in GCs and COCs EVs. A bioinformatics analysis of the identified cell‐specific and differentially expressed miRNAs predicted that they likely modulate important cellular processes, including signalling pathways such as the PI3K‐Akt, MAPK and Wnt pathways. By investigating the origins of miRNAs within the follicular fluid, the results of this study provide novel insights into follicular miRNA content and intercellular communication that may be of invaluable use in the context of reproductive technologies, diagnostic of ovarian‐related diseases and/or the identification of biomarkers for oocyte and embryo quality.     

Effect of hCG on Early Luteal Serum Progesterone Concentrations in PG600‐Treated Gilts

Gilt oestrus and ovulation responses to injection of a combination of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) (PG600) can be unpredictable, possibly reflecting inadequate circulating LH activity. The objective of this study was to determine the effect of PG600 followed by supplemental hCG on gilt ovarian responses. In experiment 1, 212 Hypor gilts (160 day of age) housed on two farms in Spain received intramuscular (i.m.) injections of PG600 (n = 47), or PG600 with an additional 200 IU hCG injected either concurrently (hCG‐0; n = 39), or at 24 h (hCG‐24; n = 41) or 48 h (hCG‐48; n = 45) after PG600. A further 40 gilts served as non‐injected controls. Ovulation responses were determined on the basis of initial blood progesterone concentrations being <1 ng/ml and achieving >5 ng / ml 10 d after the PG600 injection. The incidence of ovulating gilts having progesterone concentrations >30 ng/ml were recorded. During the study period, 10% of control gilts ovulated whereas 85–100% of hormone‐treated gilts ovulated. There were no significant differences among hormone groups for proportions of gilts ovulating. The proportions of gilts having circulating progesterone concentrations >30 ng/ml were increased (p ≤ 0.02) in all hCG treated groups compared with the PG600 group. In experiment 2, a total of 76 Hypor gilts at either 150 or 200 days of age were injected with PG600 (n = 18), 400 IU eCG followed by 200 IU hCG 24 h later (n = 20), PG600 followed by 100 IU hCG 24 h later (n = 17), or 400 IU eCG followed by 300 IU hCG 24 h later (n = 21). Blood samples were obtained 10 days later for progesterone assay. There were no effects of treatment or age on incidence of ovulation, but fewer 150‐day‐old gilts treated with PG600 or 400 IU eCG followed by 200 IU hCG had progesterone concentrations >30 ng / ml. We conclude that hCG treatment subsequent to PG600 treatment will generate a higher circulating progesterone concentration, although the effect is not evident in older, presumably peripubertal, gilts. The mechanism involved and implications for fertility remain to be determined.     

Penny ice cap cores, baffin island, canada, and the wisconsinan foxe dome connection: two states of hudson bay ice cover

Ice cores from Penny Ice Cap, Baffin Island, Canada, provide continuous Holocene records of oxygen isotopic composition (delta18O, proxy for temperature) and atmospheric impurities. A time scale was established with the use of altered seasonal variations, some volcanic horizons, and the age for the end of the Wisconsin ice age determined from the GRIP and GISP2 ice cores. There is pre-Holocene ice near the bed. The change in delta18O since the last glacial maximum (LGM) is at least 12.5 per mil, compared with an expected value of 7 per mil, suggesting that LGM ice originated at the much higher elevations of the then existing Foxe Dome and Foxe Ridge of the Laurentide Ice Sheet. The LGM delta18O values suggest thick ice frozen to the bed of Hudson Bay.     

The structure of the upper atmosphere of mars: In situ accelerometer measurements from mars global surveyor

The Mars Global Surveyor (MGS) z-axis accelerometer has obtained over 200 vertical structures of thermospheric density, temperature, and pressure, ranging from 110 to 170 kilometers, compared to only three previous such vertical structures. In November 1997, a regional dust storm in the Southern Hemisphere triggered an unexpectedly large thermospheric response at mid-northern latitudes, increasing the altitude of thermospheric pressure surfaces there by as much as 8 kilometers and indicating a strong global thermospheric response to a regional dust storm. Throughout the MGS mission, thermospheric density bulges have been detected on opposite sides of the planet near 90 degreesE and 90 degreesW, in the vicinity of maximum terrain heights. This wave 2 pattern may be caused by topographically-forced planetary waves propagating up from the lower atmosphere.     

Ingestion of artificial diets with different textures as determined by the inert marker ytterbium oxide during culture of early‐stage phyllosoma of the spiny lobster,Jasus edwardsii

Instar 1 and 2 Jasus edwardsii (Decapoda; Palinuridae) phyllosoma larvae were fed an artificial diet formulation (based on fish and mussel) consisting of three different gelatin combinations (firm, 4% inclusion; medium, 2% inclusion; and soft, no gelatin). The diet contained ytterbium oxide (Yb₂O₃; 1% inclusion), an inert heavy metal marker, which was used to confirm diet consumption in instar 1 and 2 phyllosoma. Over a 6‐h feeding period, instar 1 phyllosoma showed a preference for the soft and medium diets as opposed to the firm diet, but this effect was not significant with instar 2. The artificial diets remained palatable over the duration of the experiments, suggesting leaching did not influence diet attractiveness. It is suggested that diets comprising softer food items such as microencapsulated pellets or gelatinous formulated diets and pastes are most suitable for early‐instar phyllosoma. Furthermore, it is also suggested that Yb₂O₃ is a potentially useful marker for ongoing quantitative work on diet utilization in lobster larvae.     

The Analgesic Effects of Administering Fentanyl or Medetomidine in the Lumbosacral Epidural Space of Cats

The analgesic effects of fentanyl (4 μg/kg) and medetomidine (10 μg/kg) in 1 mL saline injected epidurally were measured in 15 cats. The response to an electrical cutaneous stimulus from a constant current generator was used as the index of analgesia. The stimulus was applied to a forelimb before epidural injection, and at 15, 30, 60, 90, 120, 180, 240, and 300 minutes post-injection (PI). The hindlimb was tested 5 minutes later. One mL saline only was used to control for volume of injection and saline. Medetomidine significantly increased the pain threshold for the hindlimb at 20 to 245 minutes PI compared with the preinjection level. Fentanyl significantly increased the pain threshold at 20 minutes PI only compared with preinjection levels. Medetomidine significantly increased the pain threshold of the forelimb at 15 to 120 minutes PI compared with the preinjection levels. Fentanyl did not significantly increase the pain threshold of the forelimb. Administration of medetomidine produced emesis in 12 of 15 cats in an average of 6.4 minutes PI (range, 3 to 11 minutes) and mild sedation in all cats. Injection of fentanyl produced no visible side effects in any of the cats.     

Comparison of tiletamine-zolazepam-ketamine and tiletamine-zolazepam-ketamine-xylazine anaesthesia in sheep

Summary The anaesthetic effects of intravenous tiletamine-zolazepam 6.6 mg/kg-ketamine 6.6 mg/kg (TK) and tiletamine-zolazepam 6.6 mg/kg-ketamine 6.6 mg/kg-xylazine 0.11 mg/kg (TKX) were evaluated in six wethers. Heart rate, respiration rate, arterial blood pressure, and the electrocardiogram were monitored during anaesthesia. Analgesia was tested by electrical stimulation in the left flank. Atropine (0.03 mg/kg) was given intramuscularly before induction, but after recording of baseline heart rate and respiratory rate. The duration of analgesia was 28.7 ± 6.9 min with TK and 82.8 ± 26.6 min with TKX. Heart rate increased significantly within 5 min after TK or TKX administration. Respiratory rate remained unchanged after TK administration, but increased significantly from 5 to 45 min after TKX administration. Arterial blood pressure decreased significantly at 15 min with TK and 30 min with TKX. Sheep remained recumbent for 201 min with TK and 166 min with TKX. All recovered uneventfully. We conclude that either TK or TKX may be used for anaesthetising sheep.     

Snake envenomation in cats and its detection by rapid immunoassay

Objective To determine the usefulness of a snake venom detection kit (SVDK) in the management of envenomed cats.
Design A clinical study.  

Animals

Twenty-two cats were investigated.
Procedure Cats injected subcutaneously with approximately 0.25 or 1.0 lethal dose (LD) of tiger snake venom or 1 or 4 LD of brown snake venom were observed for clinical symptoms of envenomation at intervals over the ensuring 24 to 48 hours(h). Blood and urine samples were taken at regular intervals and assayed in a quantitative laboratory assay for snake venoms. Selected samples were assayed in parallel in a rapid, semi-quantitative SVDK.
Results The studies showed that it was important to estimate the elapsed time from envenomation to presentation. If this time was less than 8 h, blood was the most appropriate sample and a negative result should exclude serious envenomation. If the elapsed time exceeded 8 h, it was essential that urine be sampled. Venom levels in urine were high at 8 h and approached the level of test sensitivity over 24 to 48 h; however by this time clinical signs were obvious in endangered cats.  

Conclusions

Careful use of the SVDK is a valuable aid in the management of a potentially envenomed cat.