The influence of ovulation number and ovarian dimensions on embryo production in superovulated cattle

Twenty-one cycling Angus heifers and five Holstein cows received a subcutaneous (SC) injection of 50 mg of progesterone (P) in oil for 14 consecutive days. On day 6 of (P) treatment, animals were injected intramuscularly (IM) with 6 mg of estradiol valerate, and on day 13, received an IM injection of 2,000 IU of Pregnant Mare Serum Gonadotropin. Three additional Angus heifers were used as non-hormone treated controls. Seventeen of 21 heifers and 4 of 5 cows (81%) exhibited estrus within 48 to 132 hr following P treatment. Two of the five animals in which estrus was not observed were palpated as pregnant and discarded from the study. Treatment animals showing estrus were randomly assigned either to Group I, animals bred by natural service, or Group II, animals artificially inseminated with two straws of frozen semen at 12-hr intervals for a total of four breedings. Twenty-one animals were slaughtered 2 to 6 days after the onset of estrus, and those animals in which estrus was not detected were slaughtered 10 days after the last P injection. Two of the 24 treated animals had no ovulations. A total of 397 ovulation points ($\text{397}\text{22}$) were counted for a mean ovulation rate of 18 ovulations per animal. One hundred and fifty-six ova were recovered ($\text{156}\text{397}$) for a collection rate of 39%. Group I animals had 44 of 66 (67%) of their ova fertilized while 23 of 71 (32%) of the ova in Group II were fertilized. Nineteen unfertilized eggs were collected from the three animals not observed in estrus. No differences in fertilization rates between the Group I and Group II animals were found. Mean ovarian width, length and weight in the treated animals was measured and found to be 3.5 ± 1.1 cm, 4.8 ± 1.4 cm, and 21.7 ± 21.2 gm, respectively. Ovarian width, length and weight were all positively correlated with the number of ovulations per ovary r=.74, r=.74, and r=.55, respectively. No significant correlation existed between ovarian width (r=.16), lenght (r=.21), or weight (r=.13) when compared to ova recovery rate. This result suggests that ovarian size or weight may not be the limiting factor involved in embryo recovery.     

Superovulation of cattle with equine pituitary extract and porcine ESH

Superovulation has been practiced in cattle for more than 50 years but the results have been highly variable. Scientists at six locations compared a horse pituitary extract (HAP) with a single batch of porcine FSH (pFSH) to determine the efficacy of these hormones to induce superovulation and to test for variability in the superovulatory response. Acetone-dried equine pituitaries were suspended in 40% ethanol containing 6% ammonium acetate, and the supernatant was mixed with 2.5 volumes of cold ethanol. The resulting precipitate was washed with cold ether and dried. Total doses of 18 mg of HAP and 36 mg of pFSH were injected intramuscularly (i.m.) over 4 days, two injections per day, and prostaglandin (PGF(2)alpha; 25 mg, i.m.) was administered on Day 3. Injections were begun on Days 6 to 13 of the estrous cycle. The overall ovulation rates (mean +/- SEM) for HAP and FSH were 8.8 +/- 0.7 and 15.1 +/- 1.0, respectively (n=231; P<0.01). Location interacted (P<0.01) with the type of gonadotropin for the ovulation rate. When expressed as a proportion of the number of corpora lutea, the total number of embryos recovered was greater (P=0.03) for pFSH than for HAP, but there was no difference in the number of Quality 1 and 2 embryos. The results show that HAP can induce a satisfactory superovulatory response, but there was no evidence of reduced variability of response to HAP compared with pFSH.     

Sodium/potassium adenosine triphosphatase alpha-subunit and alpha-subunit mRNA levels in early rabbit embryos

Sodium/potassium adenosine triphosphatase (Na+/K+ ATPase) and Na+/K+ ATPase mRNA content of rabbit embryos during preimplantation development were evaluated. Changes in Na+/K+ ATPase alpha-subunit content were detected with Western blotting using polyclonal antiserum against guinea pig Na+/K+ ATPase. Total RNA samples from rabbit embryos were analyzed by using Northern blots hybridized with random primer-labeled cDNA for Na+/K+ ATPase alpha-subunit from sheep kidney. Northern blots exhibited a single mRNA band (3.65 kilobases) in sheep kidneys and rabbit embryos. Between Day 4 and Day 6 of development, Na+/K+ ATPase alpha-subunit mRNA content increased 35-fold whereas Na+/K+ ATPase alpha-subunit content increased 22-fold. The similar increase in Na+/K+ ATPase alpha-subunit mRNA and alpha-subunit content in rabbit embryos suggests that Na+/K+ ATPase is partly regulated at the mRNA level during blastocyst expansion.     

Exercise training in childhood-onset systemic lupus erythematosus: a controlled randomized trial

Introduction

Exercise training has emerged as a promising therapeutic strategy to counteract physical dysfunction in adult systemic lupus erythematosus. However, no longitudinal studies have evaluated the effects of an exercise training program in childhood-onset systemic lupus erythematosus (C-SLE) patients. The objective was to evaluate the safety and the efficacy of a supervised aerobic training program in improving the cardiorespiratory capacity in C-SLE patients.

Methods

Nineteen physically inactive C-SLE patients were randomly assigned into two groups: trained (TR, n = 10, supervised moderate-intensity aerobic exercise program) and non-trained (NT, n = 9). Gender-, body mass index (BMI)- and age-matched healthy children were recruited as controls (C, n = 10) for baseline (PRE) measurements only. C-SLE patients were assessed at PRE and after 12 weeks of training (POST). Main measurements included exercise tolerance and cardiorespiratory measurements in response to a maximal exercise (that is, peak VO₂, chronotropic reserve (CR), and the heart rate recovery (ΔHRR) (that is, the difference between HR at peak exercise and at both the first (ΔHRR1) and second (ΔHRR2) minutes of recovery after exercise).

Results

The C-SLE NT patients did not present changes in any of the cardiorespiratory parameters at POST (P > 0.05). In contrast, the exercise training program was effective in promoting significant increases in time-to-exhaustion (P = 0.01; ES = 1.07), peak speed (P = 0.01; ES = 1.08), peak VO₂ (P = 0.04; ES = 0.86), CR (P = 0.06; ES = 0.83), and in ΔHRR1 and ΔHRR2 (P = 0.003; ES = 1.29 and P = 0.0008; ES = 1.36, respectively) in the C-SLE TR when compared with the NT group. Moreover, cardiorespiratory parameters were comparable between C-SLE TR patients and C subjects after the exercise training intervention, as evidenced by the ANOVA analysis (P > 0.05, TR vs. C). SLEDAI-2K scores remained stable throughout the study.

Conclusion

A 3-month aerobic exercise training was safe and capable of ameliorating the cardiorespiratory capacity and the autonomic function in C-SLE patients.

Trial registration

NCT01515163.     

Automated Entire Thrombus Density Measurements for Robust and Comprehensive Thrombus Characterization in Patients with Acute Ischemic Stroke

Background and Purpose

In acute ischemic stroke (AIS) management, CT-based thrombus density has been associated with treatment success. However, currently used thrombus measurements are prone to inter-observer variability and oversimplify the heterogeneous thrombus composition. Our aim was first to introduce an automated method to assess the entire thrombus density and then to compare the measured entire thrombus density with respect to current standard manual measurements.

Materials and Method

In 135 AIS patients, the density distribution of the entire thrombus was determined. Density distributions were described using medians, interquartile ranges (IQR), kurtosis, and skewedness. Differences between the median of entire thrombus measurements and commonly applied manual measurements using 3 regions of interest were determined using linear regression.

Results

Density distributions varied considerably with medians ranging from 20.0 to 62.8 HU and IQRs ranging from 9.3 to 55.8 HU. The average median of the thrombus density distributions (43.5 ± 10.2 HU) was lower than the manual assessment (49.6 ± 8.0 HU) (p<0.05). The difference between manual measurements and median density of entire thrombus decreased with increasing density (r = 0.64; p<0.05), revealing relatively higher manual measurements for low density thrombi such that manual density measurement tend overestimates the real thrombus density.

Conclusions

Automatic measurements of the full thrombus expose a wide variety of thrombi density distribution, which is not grasped with currently used manual measurement. Furthermore, discrimination of low and high density thrombi is improved with the automated method.     

Fire ants: What do rural and urban areas show us about occurrence, diversity,and ancestral state reconstruction?

In South America, Solenopsis saevissima and S. invicta are the most common fire ants. Nests are founded in areas under anthropic interference like urban or rural areas, but S. invicta is found preferentially in those with the greatest anthropic interference. However, we do not know the rates at which they exist in anthropized areas next to high density of native vegetation. Areas with 60 to 90% of native Atlantic Forest were selected to verify the occurrence of both species in rural and urban areas. We investigated the molecular diversity and applied the reconstruction of the ancestral state analysis for each species. A total of 186 nests were analyzed and we found that the two species had the same proportion in the urban area. However, S. saevissima had a higher rate of prevalence in the rural area, in addition to having a greater number of haplotypes and ancestry associated with this type of habitat for the region. S. invicta had the same number of haplotypes in both rural and urban regions, and less haplotypic diversity. We conclude that S. saevissima is a species typically associated with rural areas and S. invicta, although present, is not dominant in urban areas. Keywords: Biodiversity, haplotype diversity, DNA barcoding, mtDNA     

Survival of oocytes recovered from vitrified sheep ovarian tissues

The objective of this work was to develop an effective vitrification technique for cryopreserving oocytes in sheep ovarian tissues. Ovaries were surgically recovered from 15 pubertal ewes and the ovarian cortex was cut into sections. Ovarian tissues were placed in equilibration medium consisting of 4% (v/v) ethylene glycol (EG) and 20% (v/v) FBS in TCM-199 on ice for 30 min and transferred to vitrification solution (35% EG, 5% polyvinylpyrrolidone, 0.4M trehalose and 20% FBS in TCM-199) for 5 min. Ovarian tissues were vitrified by dropping the tissue on the surface of a steel cube cooled by liquid nitrogen. Cumulus-enclosed oocyte complexes (COC) were also collected and vitrified following the procedure used for ovarian tissues. After 2-3 weeks of storage in liquid nitrogen, ovarian tissues and COC were thawed at 37 degrees C in 0.3M trehalose and COC in ovarian tissues were mechanically and enzymatically isolated. Vitrified COC and freshly collected COC were washed twice in maturation medium (TCM-199 supplemented with 0.255 mM pyruvate and 10% heat-treated estrus cow serum) and cultured in 50 microl drops of maturation medium under paraffin oil for 23-25h at 39 degrees C in a humidified atmosphere of 5% CO(2) in air. After culture, cumulus cells were removed by hyaluronidase treatment and vortexing and oocytes were fixed and stained. No significant differences were observed between vitrified oocytes, oocytes recovered from vitrified ovarian tissues and non-vitrified control oocytes in the percentage of oocytes with acceptable staining per total number of oocytes fixed or with visible chromatin per total number of oocytes with acceptable staining. However, fewer (P<0.05) oocytes obtained from vitrified ovarian tissues (70%) reached metaphase II compared to vitrified oocytes (88%) and non-vitrified control oocytes (90%). In contrast, when oocytes with at least 3-5 layers of cumulus cells were considered from each of the three groups, no differences (P>0.05) were observed due to treatment in the percentages of oocytes developing to metaphase II. These results demonstrate that sheep oocytes can be successfully cryopreserved by vitrification of ovarian tissues and exhibit in vitro maturation rates similar to that of vitrified and non-vitrified oocytes.