314 RELATIONSHIP BETWEEN SERUM ANTI-MULLERIAN HORMONE (AMH), OVARIAN RESERVE, AND EMBRYO PRODUCTION IN SUPEROVULATED HOLSTEIN COWS

2013 ◽  
Vol 25 (1) ◽  
pp. 304 ◽  
Author(s):  
A. Rozner ◽  
J. Verstegen
Keyword(s):  
Embryo Production ◽  
Blood Samples ◽  
Positive Correlation ◽  
Embryo Recovery ◽  
Multiple Ovulation ◽  
Oocyte Collection ◽  
Significant Difference ◽  
Collection Data ◽  
Over Time

The relations between serum anti-Mullerian hormone (AMH), oocyte numbers, and in vivo embryo production in Holstein heifers were evaluated. The AMH levels of 15 unstimulated cows were followed at weekly intervals during their oestrous cycles and monthly for 4 months. Forty-one superovulated heifers were evaluated at ovum pick-up (OPU) performed 20 h after cystorelin administration, and 125 others were evaluated at embryo recovery. Animals were followed over 3 consecutive cycles induced using a modified Ovsynch protocol with 4 days of FSH (Pluset H, Minitube of America, Verona, WI, USA). Blood samples were collected in serum tubes and spun within 2 h. The samples were stored at –20°C until evaluation using the Minitube of America AMH-bovine specific immunoassay (AMH Fertility Assay™). The statistical analyses (ANOVA and data correlation) were performed using Statview 5 with P < 0.05. Serum AMH ranged from 43 to 960 pg mL–1. The average AMH level of all cows was stable during the oestrous cycle and for each of the 4 monthly measurements. There was a high correlation between all values per animal (r2 = 0.9077; P < 0.01), suggesting that AMH levels are consistent during the cycle and for at least 4 consecutive months. Animals that were repeatedly stimulated showed decreasing AMH levels (509 ± 295, 299 ± 210, 211 ± 119) and a decrease in recovered embryos (7.4 ± 4, 5.6 ± 3.8, 4.2 ± 3.2; P = 0.02). The number of oocytes was not altered by multiple stimulations (10.4 ± 9.8, 11.3 ± 6.2, 8.5 ± 7.6; P = 0.75). As AMH and embryo numbers decreased after multiple stimulations, only the first AMH value and results of the first OPU or flush were used to establish following correlation. Serum AMH showed a positive correlation to the number of oocytes (r2 = 0.245) and embryos collected (r2 = 0.27).When separated into AMH categories, low (<100), normal (100–400), and high (>400 pg mL–1), high-AMH OPU animals yielded significantly higher numbers of oocytes than the animals in the normal or low AMH groups (13.8 ± 9.2 v. 9.2 ± 5.2 and 5.6 ± 3.9; P = 0.001). Flushed animals with high AMH levels had significantly higher numbers of embryos than those with low AMH (10.9 ± 7.9 v. 5.7 ± 5; P = 0.002). Comparison of the first AMH value to the average number of oocytes or embryos collected over the course of 3 collections/animal showed a positive correlation to the average number of oocytes/collection from individual OPU donors (r2 = 0.436) and a positive correlation to the average number of embryos/collection from individual donors (r2 = 0.176). When separated into AMH groups, high-AMH flushed animals had significantly higher numbers of embryos than the normal- or low-AMH animals (9.3 ± 3.1 v. 5.7 ± 3.4 and 4.5 ± 2; P = 0.0001). As OPU animals with low AMH were used only once, average oocyte/collection data was not available for this category. A significant difference was observed between the high- and normal-AMH categories (12 ± 3.6 v. 7 ± 2; P = 0.0001). Circulating AMH is stable over time in unstimulated animals but decreases in repetitively stimulated animals. Anti-Mullerian hormone is highly associated with superovulation response and oocyte and embryo production, and its use should improve animal selection to achieve improve efficiency of multiple-ovulation embryo transfer.

scholarly journals Predicting chromosome damage in astronauts participating in international space station missions

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alan Feiveson ◽  
Kerry George ◽  
Mark Shavers ◽  
Maria Moreno-Villanueva ◽  
Ye Zhang ◽  
...  
Keyword(s):  
International Space Station ◽  
Dose Response ◽  
Ex Vivo ◽  
Space Station ◽  
Space Radiation ◽  
Blood Samples ◽  
International Space ◽  
Significant Difference ◽  
Subject Specific

AbstractSpace radiation consists of energetic protons and other heavier ions. During the International Space Station program, chromosome aberrations in lymphocytes of astronauts have been analyzed to estimate received biological doses of space radiation. More specifically, pre-flight blood samples were exposed ex vivo to varying doses of gamma rays, while post-flight blood samples were collected shortly and several months after landing. Here, in a study of 43 crew-missions, we investigated whether individual radiosensitivity, as determined by the ex vivo dose–response of the pre-flight chromosome aberration rate (CAR), contributes to the prediction of the post-flight CAR incurred from the radiation exposure during missions. Random-effects Poisson regression was used to estimate subject-specific radiosensitivities from the preflight dose–response data, which were in turn used to predict post-flight CAR and subject-specific relative biological effectiveness (RBEs) between space radiation and gamma radiation. Covariates age, gender were also considered. Results indicate that there is predictive value in background CAR as well as radiosensitivity determined preflight for explaining individual differences in post-flight CAR over and above that which could be explained by BFO dose alone. The in vivo RBE for space radiation was estimated to be approximately 3 relative to the ex vivo dose response to gamma irradiation. In addition, pre-flight radiosensitivity tended to be higher for individuals having a higher background CAR, suggesting that individuals with greater radiosensitivity can be more sensitive to other environmental stressors encountered in daily life. We also noted that both background CAR and radiosensitivity tend to increase with age, although both are highly variable. Finally, we observed no significant difference between the observed CAR shortly after mission and at > 6 months post-mission.

scholarly journals In vivo ultrasound guided transvaginal oocyte collection in the cow

2018 ◽  
Vol 49 (3) ◽  
pp. 195
Author(s):  
G. S. AMIRIDIS (Γ.Σ. ΑΜΟΙΡΙΔΗΣ) ◽  
M. SALAHEDDINE ◽  
I. A. JEFFCOATE ◽  
E. VAINAS (Ε. ΒΑΪΝΑΣ) ◽  
L. ROBERTSON
Keyword(s):  
Recovery Rate ◽  
Transvaginal Ultrasound ◽  
Oestrous Cycle ◽  
Ultrasound Guided ◽  
Follicular Aspiration ◽  
Oocyte Recovery ◽  
Oocyte Collection ◽  
Significant Difference

This paper describes the results of the in vivo ultrasound guided follicular aspiration for ovum pick υρ (OPU) in the cow. Twelve non pregnant dry cows aged 4-6 years were used in this experiment. Eight cows underwent OPU during three successive oestrous cycles and another four cows were used as controls having only transvaginal ultrasound scanning of their ovaries. Oocyte collection took place three times during the luteal phase of each natural oestrous cycle (days 3-4,9-11 and 14-17). The content of 326 follicles with a diameter of 4-15mm was aspirated and 104 oocytes were collected (recovery rate 31.9% or 1.55 oocytes per cow and session). The oocyte recovery rate increased after the first three sessions (from 13.04% to 35.0%) and reached levels of υρ to 52.6%. More follicles were aspirated on days 9-11 (133 follicles 40.8%) compared to 111 (34%) follicles on days 14-17 and 82 (25%) on days 3-4) (P<0.05). The evaluation of the collected oocytes revealed that 60 oocytes (57.7%) were suitable for further in vitro manipulation. Neither the origin of the oocyte (left or right ovary) nor the stage of the oestrous cycle affected the recovery rate or the quality of the collected oocytes. There was no significant difference either in the length of the oestrous cycle between the experimental animals and the controls (21.6± 1.4 vs. 22.37±1.0 respectively), or in plasma progesterone concentration in daily collected blood samples from the animals of the two groups. The results of this study are compared to those from the international literature and to the results from endoscopical methods for oocyte recovery. The feasibility of application of this technique to projects designed to improve the genetic merit of cows is discussed.

Advances in production of embryos in vitro from juvenile and prepubertal oocytes from the calf and lamb

1997 ◽  
Vol 9 (3) ◽  
pp. 333 ◽  
Author(s):  
D. T. Armstrong ◽  
P. J. Kotaras ◽  
C. R. Earl
Keyword(s):  
Embryo Production ◽  
Vitro Fertilization ◽  
Oocyte Collection ◽  
Domestic Livestock ◽  
Normal Offspring ◽  
Calf Oocytes ◽  
Collection Methods ◽  
Fresh Transfer

The use of juvenile donors in embryo-transfer (ET) programmes offers considerable potential for accelerated genetic gain in domestic livestock through reduced generation interval. The present paper reviews recent research aimed at optimizing embryo production from oocytes collected from young calves and lambs using in vitro methods of embryo production. Emphasis is placed on criteria for donor selection, oocyte-collection methods, and hormone-stimulation methods designed to produce maximum yields of viable oocytes. In vitro fertilization (IVF) rates of calf and lamb oocytes did not differ significantly whether matured in vivo or in vitro, and rates of development to blastocyst stages in culture were similar to those observed for embryos derived from adult donors. Blastocysts produced by IVF of lamb and calf oocytes established ET pregnancies at rates of 30–45%. Pregnant recipients have reached full term and delivered normal offspring at rates similar to those expected following ET of embryos produced in vivo from superovulated donors. On the basis of current follicle-stimulation protocols, on rates of blastocyst production in vitro under optimal conditions, and on observed pregnancy rates from fresh transfer of IVF embryos, 8–10 pregnancies may be expected per oocyte collection from 10–12-week-old calves and from 6–8-week-old lambs.

240 EFFECTS OF REPEATED SUPEROVULATION ON EMBRYO YIELDS IN SOME TURKISH NATIVE GOAT BREEDS

2013 ◽  
Vol 25 (1) ◽  
pp. 268
Author(s):  
M. Kaymaz ◽  
A. R. Agaoglu ◽  
K. Karakas ◽  
I. Pir Yagci ◽  
O. Korkmaz Agaoglu ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Repeated Administration ◽  
Corpora Lutea ◽  
Embryo Production ◽  
Recovery Rates ◽  
Embryo Recovery ◽  
The Third ◽  
Friedman's Test ◽  
Negative Effect

The Angora, Kilis, Honamli, Hair, and Norduz are native goat breeds in Turkey and are currently in danger of extinction. This study aimed to assess the efficacy of the repeated administration of a superovulatory (SOV) protocol for in vivo embryo production in these breeds. A total of 14 Angora, 15 Kilis, 10 Honamli, 10 Hair, and 9 Norduz goats were used in this work. The synchronization procedure was started on Day 5 after visible oestrus by using controlled internal drug release dispensers (CIDR®) for 11 days. Administration of FSH (Folltropin®) began on Day 9 (twice daily) and continued for 3 days (total dose: 200 mg; 50 mg × 2.30 mg × 2.20 mg × 2). A dose of prostaglandin F2α (1.6 mg; Dalmazin®) was injected together with first FSH injection. Gonadotropin-releasing hormone (Receptal®: 100 µg) was injected 6 h before mating. All goats in oestrus were naturally mated twice a day for 3 days. Ovarian examination (number of corpora lutea) and embryo recovery were performed by laparotomy on Day 6 after CIDR® withdrawal. Each uterine horn was flushed, and the embryos were recovered and counted. To avoid intra-abdominal adhesions, a 2.5% heparin solution was used during flushing. The SOV procedure was repeated once per year during the breeding season (2009 to 2011). Fertilization and recovery rates were calculated. Differences in the SOV response and embryo yields were evaluated by Friedman’s test. In Hair goats, the number of corpora lutea decreased significantly (P < 0.05) during the third SOV cycle (12.7 ± 6.2, 14.0 ± 9.1, and 6.8 ± 5.6, respectively, for cycles 1, 2, and 3), whereas no effect of the cycle was observed in the remainder of breeds. The number of expanded blastocysts increased considerably during the third cycle in Angora (0, 0.2 ± 0.8, and 1.4 ± 2.9, respectively, for cycles 1, 2, and 3), Kilis (0.2 ± 0.4, 0.3 ± 1.3, and 4.2 ± 5.0), and Honamli (0, 1.3 ± 1.7, and 3.6 ± 4.5) goats, whereas a significant decrease was observed in Norduz goats (2.4 ± 5.0, 1.8 ± 2.0, and 0.1 ± 0.3; P < 0.05). The mean numbers of unfertilized oocytes were found to be significantly increased in Angora (0.4 ± 1.6, 0, and 2.1 ± 4.1, respectively, for cycles 1, 2, and 3), Kilis (0, 1.3 ± 3.9, and 3.1 ± 5.2), and Honamli (0, 4.9 ± 5.2, and 4.5 ± 7.8) goats (P < 0.05). As a result, fertilization rates (%) showed a decrease in Angora (50, 100, 24.5, respectively, for cycles 1, 2, and 3) and Honamli (100, 42.5, and 56.3) goats (P < 0.05), whereas recovery rates showed no difference among the different breeds. The methodology presented in this study was found to be an efficient technique for superovulation of the Angora, Kilis, and Honamli goats. Because Hair and Norduz are relatively small breeds, the dosage of FSH might have had a negative effect on the superovulation and embryo yield. Additionally, the use of intra-abdominal washing solutions for preventing adhesions as observed in previous works (data not shown) is believed to have a positive effect on achieving high levels of efficiency in in vivo embryo production.

scholarly journals Force decay evaluation of latex and non-latex orthodontic intraoral elastics: in vivo study

2018 ◽  
Vol 23 (6) ◽  
pp. 42-47 ◽  
Author(s):  
Daniela Ferreira de Carvalho Notaroberto ◽  
Mariana Martins e Martins ◽  
Maria Teresa de Andrade Goldner ◽  
Alvaro de Moraes Mendes ◽  
Cátia Cardoso Abdo Quintão
Keyword(s):  
Clinical Study ◽  
Testing Machine ◽  
Material Degradation ◽  
Force Decay ◽  
Significant Difference ◽  
Post Hoc ◽  
Zero Time ◽  
Paired T Test ◽  
Over Time

ABSTRACT Objective: This clinical study was conducted in order to evaluate force decay over time of latex and non-latex orthodontic intraoral elastics. Methods: Patients (n = 15) were evaluated using latex and non-latex elastics in the periods of : 0, 1, 3, 12 and 24 hours. The rubber bands were transferred to the testing machine (EMIC DL-500 MF), and force values were recorded after stretching the elastic to a length of 25mm. Paired t test was applied and analysis of variance (ANOVA) was used to evaluate the variation of force generated. LSD (Fisher’s least significant difference) post-hoc test was thus employed. Results: As regards the initial forces (zero time), the values of force for non-latex elastic were slightly higher than for the latex elastic. In the subsequent times, the forces generated by the latex elastic showed higher values. Regarding the material degradation, at the end of 24 hours the highest percentage was observed for non-latex elastic. Conclusions: The latex elastics had a more stable behavior during the studied period, compared with non-latex.

In vivo embryo recovery vs. in vivo oocyte recovery and in vitro embryo production: Embryo yields and pregnancy rates in cattle

1997 ◽  
Vol 47 (1) ◽  
pp. 364 ◽  
Author(s):  
K.L. Goodhand ◽  
R.G. Watt ◽  
M.E. Staines ◽  
L.C. Higgins ◽  
D.F. Dolman ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Embryo Production ◽  
Embryo Recovery ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

scholarly journals Effects of alfaprostol and luprostiol on the embryo production within MOET technique in Black Bengal goats

1970 ◽  
Vol 2 (2) ◽  
pp. 147-150 ◽  
Author(s):  
MI Faruk ◽  
BZ Fatema ◽  
FY Bari ◽  
MGS Alam
Keyword(s):  
Treatment Group ◽  
Embryo Transfer ◽  
Treated Group ◽  
Embryo Production ◽  
Recovery Method ◽  
Treatment Groups ◽  
Multiple Ovulation ◽  
Significant Difference ◽  
Surgical Recovery ◽  
The Mean

The effects of Alfaprostol and Luprostiol on embryo production within multiple ovulation and embryo transfer (MOET) technique were studied on 16 Black Bengal goats during the period from January 2002 to June 2003. These 16 goats were randomly divided into two equal groups (A & B), each consisting of 8 goats. Each of the 16 goats was flushed in different times within MOET technique to determine the effects of alfaprostol and luprostiol on embryo production. Each group consisting of 8 donors was synchronized with alfaprostol (Gabbrostim®, VETEM, Italy) or luprostiol (Prosolvin®, Intervet International, Netherlands) @ 2 mg and 7.5 mg, equivalent to 1 ml / donor respectively. The donor goats were hand mated following the onset of oestrus, 1-2 times at 6 h interval depending on the duration of oestrus. The embryos were collected at Day 7 of mating using surgical recovery method. The mean number of ovulation in alfaprostol and luprostiol group was 8.50 ± 0.90 and 8.1 ± 0.76, respectively, where in both cases 900 iu PMSG was used for induction. There was no significant (p > 0.05) difference between this two groups on superovulation rate. The mean numbers of recovered, fertilized and transferable embryos were 5.4 ± 0.80 and 5.1 ± 0.61; 3.9 ± 0.52 and 2.6 ± 0.37 and 3.6 ± 1.6 and 2.4 ± 1.0, respectively, in alfaprostol and luprostiol treatment group. Like superovulation, there was no difference between the two treatment groups on recovered, fertilized and transferable embryos. The percentage of recovered, fertilized and transferable embryos were 63 ± 7.7 and 63 ± 3.74, 72 ± 4.55 and 51 ± 7.0 and 93.6 ± 1.6 and 90.48 ± 1.0 in alfaprostol and luprostiol treated groups, respectively. The significant difference was only existed in the percentages of fertilized embryos between the two treatment groups, where alfaprostol treated group had the significantly (p < 0.01) higher percentage of fertilized embryos.Key words: Alfaprostol; luprostiol; effect; embryo transfer; ovulation; Black Bengal goatsdoi: 10.3329/bjvm.v2i2.2558Bangl. J. Vet. Med. (2004). 2 (2): 147-150

87 OVARIAN RESERVE, EMBRYO PRODUCTION, AND THEIR CORRELATION WITH ANTI-MÜLLERIAN HORMONE (AMH) IN HOLSTEIN COWS

2015 ◽  
Vol 27 (1) ◽  
pp. 136
Author(s):  
J. Verstegen ◽  
A. Rozner
Keyword(s):  
Peptide Hormone ◽  
Holstein Cows ◽  
Embryo Production ◽  
Multiple Ovulation ◽  
Males And Females ◽  
Follicular Reserve ◽  
Student’S T ◽  
The Relationship ◽  
Student’S T Test

Anti-Müllerian hormone (AMH) is a small peptide hormone that has been associated with ovarian follicular reserve in humans and in some animal species including bovine. Profiles of AMH, as well as the relationship between serum AMH to oocyte number and in vivo embryo production, were evaluated in Holstein cows. AMH levels were determined in 15 unstimulated cows at monthly intervals for 4 months and in 394 male and 399 female developing Holstein animals from birth to adulthood. Also, AMH was measured in 41 heifers at the time of ovum pick-up (OPU) and 125 heifers at the time of embryo flushing. Superovulation was induced before OPU or embryo flushing using a modified Ovsynch protocol with 4 days of decreasing FSH (Pluset H®, MOFA Global, Verona, WI, USA). Blood samples were collected using serum tubes and spun within 2 h. The samples were stored at –20°C until evaluated for AMH using the AMH-Bovine specific immunoassay® (MOFA Global). AMH levels in males and females peaked at 2 months of age and then decrease as they reached adulthood. The average AMH level of adult cows was stable for each of the 4 monthly measurements, with a high correlation between all values per animal (r2 = 0.9077; P < 0.01), suggesting that AMH levels are consistent for at least 4 consecutive months. However, AMH levels were lowest during the summer months, suggesting a seasonal change in AMH secretion. Animals repeatedly ovarian stimulated showed decreasing AMH levels (509 ± 295, 299 ± 210, 211 ± 119) with subsequent stimulations. There was also a significant decrease in the number of embryos recovered (5.7 ± 4, 2.2 ± 1.9; P = 0.02); however, the number of oocytes was not altered by multiple stimulations (9.9 ± 9.8, 8.1 ± 6.2; P = 0.57). Because AMH and embryo numbers decreased after multiple stimulations, the first AMH value and results of the first OPU or embryo flush were used for the correlation of AMH to the number of oocytes or embryos. Animals were separated into 3 AMH categories: low (<100), normal (100–400), and high (>400 pg mL–1). High AMH OPU animals had significantly higher numbers of oocytes than the normal or low AMH groups (13.8 ± 9.2, 9.2 ± 5.3, 5.6 ± 3.9; P = 0.001). High AMH flushed animals had significantly higher numbers of embryos than animals with low AMH (10.9 ± 8.0, 5.7 ± 5; P = 0.002). Statistical analyses were performed using Statview 5. Differences were determined using Student's t-test; P < 0.05 was considered significant. In conclusion, AMH serum concentrations are consistent over multiple months; however, blood should not be taken for animal selection by AMH after ovarian stimulations have begun and should be interpreted with caution during the summer months. AMH is highly associated with superovulation response and oocyte and embryo production and should improve efficiency of multiple-ovulation embryo transfer.

141 SUPERIORITY OF FEMALE EMBRYO PRODUCTION SYSTEM BY IN VIVO-MATURED OOCYTE AND X-SORTED SPERM IN BROWN SWISS COWS

2014 ◽  
Vol 26 (1) ◽  
pp. 184
Author(s):  
T. Yamanouchi ◽  
H. Matsuda ◽  
M. Ohtake ◽  
K. Masaki ◽  
E. Horiguchi ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Female Embryo ◽  
Brown Swiss ◽  
Embryo Recovery ◽  
Matured Oocyte ◽  
Matured Groups

Embryo transfer using a female embryo is an effective tool for offspring production on dairy industry; however, embryo production by embryo recovery (ER) using X-sorted semen is not sufficient because non-fertilized oocytes are recovered frequently. In Holstein cows, we developed a system for high blastocyst production that was performed by IVF using X-sorted sperm and in vivo-matured oocytes obtained by ovum pickup (OPU) after superstimulation. The purpose of this study was to adjust this system to Brown Swiss cows, comparing between ER and embryo production from oocytes derived from OPU with or without superstimulation. In the ER group, cows (n = 10) received a CIDR (Day 0) and 2 mg of oestradiol-benzoate on Day 1. A total of 30 Armour Units of FSH were injected into cows twice a day, with decreasing doses from the evening of Day 5 to the morning of Day 9. On the evening of Day 7 or 8, 0.75 mg of prostaglandin F2α (cloprostenol) was injected. The CIDR was removed on Day 8 or 9 and 0.2 mg of gonadotropin-releasing hormone (GnRH; fertirelin acetate) were injected on Day 9 or 10. At oestrus, AI was carried out twice, 12 h apart, with a total of 4 straws of X-sorted semen per cow. In the OPU group, cows (n = 7) were subjected to OPU without any pretreatment, collected immature oocytes were in vivo matured for 20 to 22 h, followed by IVF using X-sorted sperm for 6 h; then, presumptive zygotes were in vitro cultured (IVC) for 9 days. In the in vivo-matured oocyte group (matured group), a CIDR was inserted (Day 0) in cows (n = 4), all follicles larger than 8 mm were removed on Day 5. Administration of FSH, prostaglandin F2α, and GnRH, as well as withdrawal of CIDR, were performed as in the ER group. In vivo-matured oocytes were collected from follicles larger than 5 mm by OPU at 25 to 26 h following GnRH injection; collected oocytes with expanded cumulus cells were fertilized with X-sorted sperm 30 h after GnRH. After 6 h of IVF, presumptive zygotes were transferred to in vitro culture, as in the OPU group. Data were compared among 3 groups; the ER group was analysed for number of CL, collected embryos, and normal embryos, against the number of aspirated follicles, collected oocytes used for IVF, and formed blastocysts in the OPU and matured groups, respectively, by Tukey-kramer test after ANOVA. There were no differences between the number of CL in the ER group and the number of follicles in the OPU and matured groups (16.4 ± 5.3 v. 31.6 ± 22.7 v. 18.5 ± 4.7, mean ± s.d., respectively). Also the number of collected embryos in the ER group and number of oocytes for IVF in the OPU and matured groups (12.8 ± 7.6 v. 14.9 ± 11.8 v. 17.8 ± 7.7, respectively) was similar. However, the number of blastocysts in the matured group (13.0 ± 5.9; P < 0.01) was higher than that in the OPU group (3.0 ± 2.2) and in the ER group (2.8 ± 3.7). For female embryo production in Brown Swiss cows using X-sorted sperm, the system of IVF with in vivo matured oocytes obtained by OPU is more effective than ER or OPU without pretreatment.

ADAMTS13 Autoantibodies, Antigen and Proteolytic Activity in Patients with Thrombotic Thrombocytopenic Purpura.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2642-2642
Author(s):  
Suresh G. Shelat ◽  
Paula Smith ◽  
Jihui Ai ◽  
X. Long Zheng
Keyword(s):  
Thrombotic Thrombocytopenic Purpura ◽  
Proteolytic Activity ◽  
Inhibitory Activity ◽  
Adamts13 Activity ◽  
Thrombocytopenic Purpura ◽  
Positive Correlation ◽  
Significant Difference ◽  
Inhibitory Antibodies

Abstract Many patients with thrombotic thrombocytopenic purpura (TTP) harbor autoantibodies against the ADAMTS13 protease, which can block proteolytic activity or accelerate clearance of this protease. The autoantibodies may be categorized as ‘inhibitory’ antibodies that bind and inhibit ADAMTS13 activity in vitro and ‘non-inhibitory’ antibodies that may bind, but do not affect the ADAMTS13 activity. The pathophysiologic role of two types of anti-ADAMTS13 autoantibodies on the level of ADAMTS13 antigen and activity remains unclear. To address this question, the relationship between the autoantibodies, and the levels of antigen and proteolytic activity in 40 patients with TTP (26 idiopathic and 14 non-idiopathic) was determined. The diagnosis of TTP was based on the presence of thrombocytopenia and microangiopathic hemolytic anemia with or without renal dysfunction and neurological symptoms. The ADAMTS13 antigen was determined by ELISA (American Diagnostica, Stamford CT). The ADAMTS13 autoantibodies were determined by Technozyme-ELISA (Technoclone GmbH, Vienna, Austria) and Immunoprecipitation and Western blotting. The ADAMTS13 activity was determined by Fluorescent resonance energy transfer (FRETS)-VWF73. The results showed that: 1) autoantibodies were present in 24 of 40 (60%) of all TTP patients, 19 of 26 (73%) in idiopathic TTP and 5 of 14 (36%) in non-idiopathic TTP. The prevalence of non-inhibitory autoantibodies was approximately 31% (8/26) in patients with idiopathic TTP and 36% (5/14) in patients with non-idiopathic TTP; 2) there was a positive correlation between the ADAMTS13 antigen levels and enzymatic activity in 36 samples tested (r=0.665, P&lt;0.0001). This positive correlation was not altered even after removing from the analysis samples demonstrating inhibitors and low activity; 3) with respect to antigen levels, there was no significant difference between idiopathic vs. non-idiopathic TTP patients, whereas ADAMTS13 activity are significantly lower in the former (Table 1); 4) the inhibitory autoantibodies significantly reduce ADAMTS13 activity and may or may not decrease ADAMTS13 antigen levels. However, non-inhibitory autoantibodies neither decrease ADAMTS13 levels nor proteolytic activity (Table 2). We conclude that overall, in TTP patient samples, ADAMTS13 antigen levels correlate with activity. Subgroup analysis indicates that idiopathic and non-idiopathic TTP patients have similar ADAMTS13 antigen levels, whereas the former demonstrates much less proteolytic activity. TTP patients with inhibitory autoantibodies may have a more profound effect on ADAMTS13 activity (and perhaps also on clearance) than those with non-inhibitory autoantibodies. The non-inhibitory antibodies do not appear to accelerate ADAMTS13 protease clearance in vivo. Table 1. ADAMTS13 Antigen and Activity in Idiopathic and Non-Idiopathic TTP Patients Patients Antigen (ng/ml) Activity (%) * p &lt; 0.05 Idiopathic 276 ± 46 (n=19) 16.9 ± 6 (n=21)* Non-idiopathic 328 ± 43 (n=17) 45.1 ± 5.7 (n=19) Table 2. Effect of Autoantibodies on ADAMTS13 Antigen and Activity Antibodies Definition Antigen (ng/ml) Activity (%) *p=0.085; **p &lt; 0.05 vs "non-inhibitory" and &lt;0.05 vs. "None" group Inhibitory Activity &lt;10%, Inhibitor (1:1), positive, TecZym&gt;15 U/ml 183 ± 55* 4.1 ± 1.2 (n=9)** Non-inhibitory Activity&gt;10%, Inhibitor (1:1), negative, TecZym &gt;15 U/ml 368 ± 52 37.6 ± 8 (n=17) None Activity &gt;10%, Inhibitor (1:1), negative, TecZym &lt;12 U/ml 294 ± 51 40.2 ± 7 (n=13)

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