Master\u27s Recital

List of performers and performances

A multicenter experience using adipose-derived mesenchymal stem cell therapy for cats with chronic, non-responsive gingivostomatitis.

BackgroundThe ability of mesenchymal stem cells (MSCs) to modulate immune responses inspired a series of clinical trials addressing oral mucosal inflammation. We previously reported on the safety and efficacy of fresh, allogeneic and autologous, adipose-derived mesenchymal stem cells (ASCs) to treat feline gingivostomatitis (FCGS), an oral mucosal inflammatory disease that shares similarities with human oral lichen planus.MethodsTo meet clinical demand and goals for future commercialization, we determined the feasibility of shipping fresh ASCs to distant clinics and extended our pilot studies to expand safety and efficacy data for shipped and non-shipped ASCs in a cohort of 18 FCGS cats enrolled locally and at a few different locations within the USA.ResultsWe found that ASCs retained their viability, phenotype, and function after shipment. ASCs administered systemically resulted in a 72% positive response rate, identical to that noted in our previous studies. Cats that responded to ASC therapy had a significant decrease in circulating globulin concentration and histological evidence of decreased CD3+ T cells and CD20+ B cells in the oral mucosa. Responder cats also had significantly decreased percentages of CD8lo cells in blood prior to and at 3 months post-ASC therapy. CD8lo cells may serve as a potential "predictor" for response to systemic ASC therapy.ConclusionFresh feline ASCs can be successfully shipped and administered to cats with FCGS. ASCs modulate the immune response and demonstrate efficacy for chronic oral mucosal inflammatory lesions that are characterized by CD8+ T cell inflammation and T cell activation. FCGS is a potentially useful naturally occurring large animal model of human oral inflammatory diseases

Allogeneic Stem Cells Alter Gene Expression and Improve Healing of Distal Limb Wounds in Horses.

Distal extremity wounds are a significant clinical problem in horses and humans and may benefit from mesenchymal stem cell (MSC) therapy. This study evaluated the effects of direct wound treatment with allogeneic stem cells, in terms of gross, histologic, and transcriptional features of healing. Three full-thickness cutaneous wounds were created on each distal forelimb in six healthy horses, for a total of six wounds per horse. Umbilical cord-blood derived equine MSCs were applied to each wound 1 day after wound creation, in one of four forms: (a) normoxic- or (b) hypoxic-preconditioned cells injected into wound margins, or (c) normoxic- or (d) hypoxic-preconditioned cells embedded in an autologous fibrin gel and applied topically to the wound bed. Controls were one blank (saline) injected wound and one blank fibrin gel-treated wound per horse. Data were collected weekly for 6 weeks and included wound surface area, thermography, gene expression, and histologic scoring. Results indicated that MSC treatment by either delivery method was safe and improved histologic outcomes and wound area. Hypoxic-preconditioning did not offer an advantage. MSC treatment by injection resulted in statistically significant increases in transforming growth factor beta and cyclooxygenase-2 expression at week 1. Histologically, significantly more MSC-treated wounds were categorized as pro-healing than pro-inflammatory. Wound area was significantly affected by treatment: MSC-injected wounds were consistently smaller than gel-treated or control wounds. In conclusion, MSC therapy shows promise for distal extremity wounds in horses, particularly when applied by direct injection into the wound margin. Stem Cells Translational Medicine 2018;7:98-108

Use of Pregnancy Associated Glycoproteins to Determine Fetal Age Throughout Gestation and Clearance Rate in Postpartum Beef Cattle

Study Description: Previously identified pregnant females from four different herds and postpartum females from one herd were utilized. Blood samples were collected (n = 1,753; study 1) between d 28 and 285 of gestation and (heifers n = 418 and cows n = 657; study 2) once a week for up to 12 weeks after calving. Serum was tested in duplicate using a commercially available blood pregnancy test, IDEXX Alertys Pregnancy Test. In study 1, procedures were modified to allow PAG concentrations to fall within the detectible range of the assay. Data were analyzed using the MIXED procedure of SAS with cow age and gestational age (also divided into four gestational age groups: 1) \u3c 30 d; 2) 30-90 d; 3) 91-180 d; 4) \u3e 180 d) in the model and then analyzed further using the REG procedure in SAS within gestational age group. In study 2, data were analyzed as repeated measure using the MIXED procedure of SAS with cow age, days postpartum (dpp), and cow age by dpp in the model, then data were analyzed further using the REG procedure in SAS. In study 1, there was a significant effect of gestational age and cow age by gestational age interaction (P \u3c 0.01) as well as a tendency of cow age (P = 0.08). Among heifers and cows, serum PAG concentrations did not differ between gestational age groups 1 and 2 (P \u3e 0.84), however, PAG concentrations differed between groups 2 and 3 (P \u3c 0.0001) and 3 and 4 (P \u3c 0.0001). There was a positive correlation between gestational age and PAG concentrations (P \u3c 0.01; r2 = 0.2604). In study 2, there was a significant effect of days postpartum (dpp; P \u3c 0.01) on PAG concentrations; however, PAG concentrations were not influenced by cow age (P = 0.73) or cow age by dpp (P = 0.55). Concentrations of PAGs rapidly decreased from d 0 to 50 postpartum and then continued to gradually decrease (P \u3c 0.01; r2 = 0.8083). Prior to 42 dpp, PAG concentrations were sufficiently elevated which resulted in false positive readings

Relationship of DAG1 and SERPINA5 Sperm Proteins With Bull Fertility

Study Description: Semen from 22 dairy bulls was used to evaluate the presence, localization, and quantification of DAG1 and SERPINA5 on sperm. Sperm motility parameters and viability was also evaluated for semen from each bull. Semen from 19 out of the 22 dairy bulls was used for in vitro embryo production (two Low-SCR and one High-SCR were not available for in vitro embryo production). Bulls were classified based on their sire conception rates (SCR) values as High-SCR (SCR \u3e 1.0) or Low-SCR fertility (SCR \u3c -4.0). Low fertility bulls were subdivided based on their blastocyst rate (BL) as High-BL (Low-SCR/High-BL BL ≥ 31%) or Low-BL (Low-SCR/Low-BL BL ≤ 26%), and High-SCR bulls were not subdivided. The GLM procedure in SAS was used with bull as a fixed effect to determine if variance was greater between bulls compared to within bull. Correlations were determined among DAG1 and SERPINA5 concentrations, percentage of tail labeled for SERPINA5, SCR, sperm total motility, progressive motility, and viability, and in vitro embryo produced cleavage rate (CL) and BL. The GLIMMIX procedure of SAS was used to evaluate the relationship of bull field fertility (High- and Low-SCR), and field and in vitro fertility (High-SCR, Low-SCR/High-BL, Low-SCR/Low-BL) classifications with sperm total (TMOT) and progressive (PROG) motility, viability, CL, BL, DAG1 and SERPINA5 relative concentration, and proportion of sperm tail labeled for SERPINA5. Both SERPINA5 and DAG1 were localized on the sperm head; however, SERPINA5 was also localized on the sperm tail. There was greater variance in concentration among bulls compared to within bull for both DAG1 (P \u3c 0.01; 69.4 vs 49.1, respectively) and SERPINA5 (P \u3c 0.01; 325.8 vs 285.4, respectively). There was a positive correlation between concentration of DAG1 and SERPINA5 (P = 0.01; r = 0.54). Concentrations of SERPINA5 were also correlated with CL (P = 0.04; r = 0.48), and percentage of sperm tail labeled for SERPINA5 was correlated with viability (P = 0.05; r = 0.44) and tended to be correlated with CL (P = 0.10; r = 0.39). There was no relationship between SCR or BL rate classifications and DAG1 (P ≥ 0.66), SERPINA5 (P ≥ 0.54), or percentage of sperm tail labeled for SERPINA5 (P ≥ 0.48)

Lipid II overproduction allows direct assay of transpeptidase inhibition by β-lactams

Peptidoglycan is an essential crosslinked polymer that surrounds bacteria and protects them from osmotic lysis. Beta-lactam antibiotics target the final stages of peptidoglycan biosynthesis by inhibiting the transpeptidases that crosslink glycan strands to complete cell wall assembly. Characterization of transpeptidases and their inhibition by beta-lactams has been hampered by lack of access to substrate. We describe a general approach to accumulate Lipid II in bacteria and to obtain large quantities of this cell wall precursor. We demonstrate utility by isolating Staphylococcus aureus Lipid II and reconstituting the synthesis of crosslinked peptidoglycan by the essential penicillin-binding protein 2, PBP2, which catalyzes both glycan polymerization and transpeptidation. We also show that we can compare the potencies of different beta-lactams by directly monitoring transpeptidase inhibition. The methods reported here will enable a better understanding of cell wall biosynthesis and facilitate studies of next-generation transpeptidase inhibitors

Influence of Bovine Viral Diarrhea Virus Infection on Artificial Insemination Conception and Breeding Season Pregnancy Success in Vaccinated Beef Herds

Bovine Viral Diarrhea Virus (BVDV) causes reproductive economic losses in cattle. The objective of this study was to evaluate the influence of BVDV infection on reproductive success. Vaccinated cows (n = 370) and heifers (n = 528) from nine different herds were synchronized using the 7-day CO-Synch + CIDR protocol and were bred using fixed-time artificial insemination (FTAI). On d 28 following insemination, blood samples were collected and pregnancy status was determined. Non-pregnant animals were resynchronized and FTAI occurred a second time. In six herds, bulls were comingled with females beginning 10-15 d after the second AI. Final pregnancy status was determined 33-80 d following the first pregnancy diagnosis. Blood samples were tested for the presence of BVDV antigen using the IDEXX BVDV PI X2 Kit. Animals that tested positive were considered infected with BVDV at the time of blood collection. Herds were determined to be BVDV infected by the presence of at least one animal having a positive test for antigen (n = 4 infected herds, n = 5 non-infected herds). Statistical analyses were performed using the GLIMMIX procedure of SAS with herd as a random variable. Herds that had evidence of BVDV infection at d 28 following insemination had significantly decreased (P \u3c 0.01) first service AI conception rates compared to herds that had no evidence of infection (34 ± 2.3% vs. 54 ± 2.3%, respectively). Additionally, breeding season pregnancy rates were decreased (P \u3c 0.01) in BVDV infected herds compared to non-infected herds (69 ± 3.4% vs. 80 ± 3.6%, respectively). There was no significant effect of BVDV infection status on embryonic loss (P = 0.42) or percentage of animals which lost a pregnancy and rebred by the end of the breeding season (P = 0.63). In conclusion, BVDV infection in well vaccinated herds had a significant negative impact on both first service AI conception rate and overall breeding season pregnancy success

The Interactions of Change in Nutrition on Uterine Environment and Cholesterol Concentrations in Beef Cattle

The objective of this study was to evaluate the impact of nutritional changes prior to and after artificial insemination (AI) on uterine environment and plasma cholesterol concentrations

Comparison of Lateral Flow to Other Pregnancy Determination Methods in Order to Determine Accuracy of Pregnancy Status in Beef Cattle Pre and Postpartum

Study Description: Blood samples were collected from six different Bos taurus herds between day 27 and 285 of gestation (heifers n = 1,205 and cows n = 1,539). Blood samples to determine PAG clearance interval were collected weekly postpartum for up to 12 weeks (heifers n = 418 and cows n = 657). Serum was tested using the lateral flow test and were read by two technicians who were blind to pregnancy status. Level of agreement between the tests were determined by Pearson’s correlation coefficients and Kappa scores. The MIXED procedure of SAS was used to evaluate the effect of dpp and age (heifers or cows) on postpartum test results. There was a positive correlation between transrectal ultrasonography and the lateral flow test (r2 = 0.71; P \u3c 0.01), and agreement between the two tests was very good (Kappa = 0.84). Of the animals that were diagnosed nonpregnant by transrectal ultrasonography, 5.61% were called pregnant by the lateral flow test. Of the animals diagnosed pregnant by transrectal ultrasonography, 2.00% were called not pregnant by the lateral flow test. Thus, a 92.38% agreement occurred between the two methods. For postpartum samples, there was no effect (P = 0.21) of age, but there was an effect of dpp (P \u3c 0.01) and a tendency for a dpp by age interaction (P = 0.06). All animals were still considered pregnant from the previous pregnancy through 35 dpp (100 ± 2.58%). The percentage of females receiving a false positive test result further decreased with time postpartum, by 77 dpp there were 13.72 ± 3.16% of the females positive for pregnancy and at 84 dpp there were 4.11 ± 4.39% positive for pregnancy detection