The study of spermatogonial stem cells (SSCs) provides a model to better understand adult stem cell biology. Besides the biomedical potential to perform studies of infertility in many species, SSCs hold a promising application at animal transgenesis. Because stem cells are thought to be associated with basement membranes, expression of α‐6 integrin has been investigated as a marker of type A spermatogonial cells, which are considered SSCs because of their undifferentiated status and self‐renewal ability. In this manner, the aim of this study was to isolate type A SSCs from adult bulls by a two‐step enzymatic procedure followed by a discontinuous Percoll density gradient purification and verify the expression of α‐6 integrin by flow cytometry and real‐time RT‐PCR before and after Percoll purification. Spermatogonial cells were successfully obtained using the two‐step enzymatic digestion. An average of 1 × 105 viable cells per gram of testis was isolated. However, the discontinuous Percoll did not purify isolated cells regarding α‐6 integrin expression. Flow cytometry analysis demonstrated no differences in the α‐6 integrin expression between cell samples before and after Percoll purification (p = 0.5636). The same was observed in the real‐time PCR analysis (p > 0.05). In addition to α‐6 integrin, the expression of GFRa‐1 and PGP9.5, known bovine SSCs markers, was detected in all samples studied. Considering that Percoll can reduce cell viability, it is possible to conclude that Percoll density gradient is not suitable to purify bovine SSC, according to α‐6 integrin expression. 相似文献
AIM: To evaluate the conception rate to fixed-time artificial insemination (FTAI) of two oestrus synchronisation programmes in dairy heifers on eight farms over 2 years.METHODS: The study was conducted in 2008 and 2010 on eight farms near Palmerston North, New Zealand. Nulliparous Friesian and Friesian×Jersey heifers (13–15 months of age) were randomly allocated to one of two oestrus synchronisation programmes. Group 1 (GPG+P4; n=330), received gonadotrophin-releasing hormone (GnRH) I/M on Day 0, a progesterone (P4)-releasing intravaginal device from Days 0–7, prostaglandin F2α (PGF) I/M on Day 7 and a second dose of GnRH at the time of FTAI on Day 9. The second group (P4+PGF; n=343) received a P4-releasing intravaginal device from Days 0–7, PGF on Day 6 and FTAI on Day 9. Pregnancy was diagnosed from Days 42–52 by transrectal ultrasonography.RESULTS: The overall conception rate was 52.4% and 54.8% for the GPG+P4 and P4+PGF groups, respectively. The odds of conception for the two treatments were not different (OR=0.90; 95% CI=0.67–1.23), nor was there any difference between groups in different years (p=0.58). Farm affected conception rate (p=0.002), but there was no interaction with treatment (p=0.92) .CONCLUSIONS: This study has shown that an alternative synchronisation programme can produce similar results in terms of conception rate to the GPG+P4 treatment, currently commonly used in heifers. More research is required to establish whether other modifications to the GPG+P4 programme can produce similar results at lower costs, and to identify and quantify farm factors which affect the economic benefit of heifer synchronisation.CLINICAL RELEVANCE: This study indicated that synchronising heifers with P4 and PGF resulted in conception rates equivalent to those resulting from a GPG+P4 treatment, but with reduced drug costs. However, because heifers in the GPG+P4 group received the second GnRH injection at the time of AI, they needed only three yardings as opposed to the four required for the heifers treated with P4 and PGF. Thus, the choice of programme for an individual farm will depend on that farm's circumstances, in particular the cost of yarding the heifers. 相似文献
Aims: To assess the inter-observer agreement for detecting bovine digital dermatitis (BDD) lesions in digital colour photographs of the hind feet of cows, which had been taken while the animals were standing to be milked, between two trained observers.
Methods: Thirty-six photographs were selected from a total of 184 photographs held by the first author (R1), who had classified them as negative (n=11) or positive (n=25) for BDD. They were delivered to a technician (R2) who had previously visually inspected cattle for BDD lesions, and who then recorded the photographs as being either BDD-positive or BDD-negative. The percentage agreement between R1 and R2, and two other inter-observer agreement statistics, Cohen’s κ and Gwet’s first-order chance correction agreement coefficient (AC1), were calculated. The cumulative membership probabilities of Cohen’s κ and Gwet’s AC1 were then calculated for different benchmark ranges of κ.
Results: The percentage agreement between R1 and R2 was 33/36 (92%), Cohen’s κ was 0.80 (95% CI=0.57–1.0) and Gwet’s AC1 was 0.86 (95% CI=0.69–1.0). Based on the cumulative membership probabilities for Gwet’s AC1, there was 75% probability that the two observers had almost perfect agreement (κ≥0.81). For both Cohen’s κ and Gwet’s AC1, there was >95% probability that the two observers had at least substantial agreement (κ≥0.61).
Conclusions: The two trained observers had at least substantial agreement in identifying from a digital photograph as to whether BDD lesions were present or absent. Therefore results from the two could be used interchangeably.
Clinical Relevance: Visual assessment for BDD lesions in the milking parlour can be subjective. However a high agreement between these two trained BDD inspectors means BDD prevalence reported from different regions in New Zealand by these two can be directly compared. 相似文献
Platinum catalysts are reported for the direct, low-temperature, oxidative conversion of methane to a methanol derivative at greater than 70 percent one-pass yield based on methane. The catalysts are platinum complexes derived from the bidiazine ligand family that are stable, active, and selective for the oxidation of a carbon-hydrogen bond of methane to produce methyl esters. Mechanistic studies show that platinum(II) is the most active oxidation state of platinum for reaction with methane, and are consistent with reaction proceeding through carbon-hydrogen bond activation of methane to generate a platinum-methyl intermediate that is oxidized to generate the methyl ester product. 相似文献
The migration and evolution of a deep ocean hydrothermal event plume were tracked with a neutrally buoyant RAFOS float. The float remained entrained in the plume for 60 days, and the plume vorticity was calculated directly from the anticyclonic motion of the float. Concentrations of suspended particles, particulate iron, and dissolved manganese in the plume did not decay significantly during the 60 days, which indicates that event plumes would be easily detectable a year after formation. 相似文献
Are extinctions of marine vertebrates as rare and unlikely as current data indicate? Long-term research surveys on the continental shelf between the Grand Banks of Newfoundland and southern New England reveal that one of the largest skates in the northwest Atlantic, the barndoor skate (Raja laevis), is close to extinction. Forty-five years ago, research surveys on St. Pierre Bank (off southern Newfoundland) recorded barndoor skates in 10% of their tows; in the last 20 years, none has been caught, and this pattern of decline is similar throughout the range of the species. 相似文献