Investigation of apoptosis in cultured cells infected with equine herpesvirus 1

Many viruses alter different stages of apoptosis of infected cells as a strategy for successful infection. Few studies have addressed mechanisms of equine herpesvirus 1 (EHV-1) strain-induced cell death. We investigated the effect of an abortigenic strain (AR8 strain) on heterologous Madin–Darby bovine kidney cells and homologous equine dermis (ED) cells cell lines. We compared morphologic and biochemical features of early and late apoptosis at different postinfection times. We investigated translocation of phosphatidylserine to the cell surface, nuclear fragmentation and changes in the cytoskeleton using flow cytometry and annexin V/propidium iodide staining, DNA laddering, terminal deoxynucleotidyl transferase UTP nick-end labeling assay and immunofluorescence staining of cytokeratin 18 cleavage. AR8 EVH-1 strain interfered with apoptosis in both cell lines, particularly during the middle stage of the replication cycle; this was more evident in ED cells. Although this antiapoptotic effect has been reported for other alpha herpesviruses, our findings may help elucidate how EHV-1 improves its infectivity during its cycle.     

Insulin-like growth factor (IGF) family genes are aberrantly expressed in bovine conceptuses produced in vitro or by nuclear transfer

Embryos produced through somatic cell nuclear transfer (NT) or in vitro production (IVP) are often associated with increased abortion and abnormalities thought to arise from disruptions in normal gene expression. The insulin-like growth factor (IGF) family has a major influence on embryonic, fetal and placental development; differences in IGF expression in NT- and IVP-derived embryos may account for embryonic losses during placental attachment. In the present study, expression of IGF-I, IGF-II, IGF-I receptor (IGF-IR), and IGF-IIR mRNAs was quantitated in Day 7 and 25 bovine embryos produced in vivo, by NT, IVP, or parthenogenesis, to further understand divergent changes occurring during development. Expression of the IGF-I gene was not detected in Day 7 blastocysts for any treatment. However, there were no differences (P>0.10) among Day 7 treatments in the amounts of IGF-IR, IGF-II, and IGF-IIR mRNA. For Day 25 conceptuses, there was higher expression of IGF-I mRNA for NT and IVP embryonic tissues than for in vivo embryonic tissues (P<0.05). Furthermore, embryonic tissues from NT-derived embryos had higher expression of IGF-II mRNA than IVP embryonic tissues (P<0.05). Placental expression of IGF-IIR mRNA was greater for NT-derived than in vivo-derived embryos (P<0.05). There were no differences in IGF-IR mRNA across all treatments and tissues (P>0.10). In conclusion, these differences in growth factor gene expression during early placental attachment and rapid embryonic growth may directly or indirectly contribute to increased losses and abnormalities in IVP- and NT-derived embryos.     

Stent implantation through a self-expanding stent

Jailing of a side-branch is a known complication of stent implantation, and makes access to the side-branch difficult, especially if the stent is of the self-expanding type. Although plain balloon angioplasty is feasible for the jailed side-branches, the use of newer devices (a stent, Rotablation or atherectomy) has not been described. We describe a novel way of treating a side-branch jailed by a self-expanding stent by using stent implantation through the strut of a self-expanding stent.     

Influence of summer heat stress on pregnancy rates of lactating dairy cattle following embryo transfer or artificial insemination

Lactating Holstein cows were used to determine if pregnancy rate from embryo transfer (n = 113) differed from contemporary control cows (n = 524) that were artificially inseminated (AI). Holstein heifers (n = 55) were superovulated with FSH-P (32 mg total) and inseminated artificially during estrus and subsequently managed under shade structures. On Day 7 post estrus, embryos were recovered, and primarily excellent to good quality embryos (90.3%) were transferred to estrus-synchronized lactating cows. Cows were managed under conditions of exposure to summer heat stress. Pregnancy status was determined by milk progesterone concentrations at Day 21 and palpation per rectum at 45 to 60 d post estrus. Pregnancy rates of cows presented for AI (Day 21, 18.0%; Days 45 to 60, 13.5%) were typical for lactating cows inseminated during periods of summer heat stress in Florida. Pregnancy rates of embryo recipient cows were higher (P<0.001) than those of control cows (Day 21, 47.6%; Days 45 to 60, 29.2%). Summer heat stress had no adverse effect on heifer superovulatory response, but it increased (P<0.05) the incidence of retarded embryos (相似文献   

Effect of timing of prostaglandin PGF 2 alpha injection subsequent to embryo collection on the resumption of normal follicular development following superovulatory treatment in cattle

Nonlactating Holstein and Jersey cows (n = 24) were superovulated and ovarian follicular development was monitored by transrectal ultrasound during the period after embryo recovery. Luteolysis was induced by two injections of prostaglandin F(2)alpha (PGF; 25 mg Lutalyse; 12-h interval) at specific times after superovulatory induced estrus (Treatment 1, Day 9; Treatment 2, Day 12; Treatment 3, Day 17; Treatment 4, Day 25; superovulatory estrus = Day 0 of Cycle 1). Follicular development was monitored during Cycle 1 before and after PGF injection and continued through the ensuing estrous cycle (Cycle 2). Superovulation led to more than one embryo collected in 14 cows (mean = 8.71 embryos: positive superovulatory response [PSR] cows), while 10 cows were not successfully superovulated (mean = 0.1 embryo; negative superovulatory response [NSR] cows). These cows differed in terms of number of unovulated follicles detected at embryo collection (4.21 vs 17.2, PSR vs NSR) and plasma progesterone during the superovulatory estrous cycle (32.3 ng/ml PSR vs 8.6 ng/ml NSR). Follicular development during Cycle 1 started sooner in NSR than in PSR cows (day by class by response P<0.03) and was initiated on Days 11 to 12 in NSR cows and on Days 19 to 20 in PSR cows. Interval to estrus after PGF averaged 6.3 d. Cows having short intervals to estrus had follicles at the time of PGF injection. Treatment influenced the length of Cycle 1, but it did not affect the interval to estrus after PGF, the length of Cycle 2, or follicular development during Cycle 2. The results indicate that 1) the timing of PGF injection after embryo collection does not influence subsequent follicular populations, 2) elongated estrous cycles and intervals to estrus after PGF in superovulated cattle are a function of decreased follicular activity, and 3) the presence of numerous corpora lutea and not the superovulatory treatment, per se, seem to attenuate follicular growth.     

Preparation of teaser bulls and steroid-implanted steers and their effectiveness in detecting estrus

Pen-O-Blocks were easily installed in 15 bulls. Libido appeared to be unaffected. However, with the exception of one bull, infections occurred, or the penis bypassed the block, or the pin broke, and in 14 tests the Pen-O-Blocks were removed within 5 weeks of installation. Recovery from infection following removal of the device was rapid and uneventful.Eight bulls had their penes deviated between 2 weeks and 4 months of age. Four of these also were castrated and implanted with androgens at 12 months of age. Semen quality and libido in the deviated bulls were normal.Bulls and steers were equipped with Chin Ball mating devices. Bulls generally were more effective than steers in detecting estrus. When placed in pens 24 hours per day with 20 heifers each, one bull detected 97% and another bull detected 100% of the potential estruses during a period of two months. An observer watching for .5 hours each morning and each late afternoon detected 74%.While steers were less effective, they were more docile than the bulls. Thus, with selection of active animals and adjustment of hormone therapy, implanted steers may be successful detectors of estrus with minimal danger to the owner. Reproductive development in the androgen-treated steers was delayed but maturation of the penis had occurred in steers slaughtered at 31 months of age. Body weight gains of steers tended to be accelerated during periods when they had implants.     

Early Mobilization after Myocardial Infarction: a Controlled Study

During the years 1968-71 203 patients with proved myocardial infarction were admitted to the trial. Patients were mobilized either on day 10 (102 patients) or on day 20 (100 patients). All patients were kept in hospital for 30 days in order to ensure a detailed comparison of clinical course and laboratory data. In neither group was there a fatal complication and the differences in clinical outcome or laboratory data were statistically not significant. Half of the patients from each group were re-examined after an average of one-and-a-half years, and again no differences were observed. It is concluded that patients with an uncomplicated myocardial infarction may safely be mobilized after 9 days and discharged after three weeks.     

Characteristics of prostaglandin F measurements in the ovarian circulation during the oestrous cycle and early pregnancy in the cow

Holstein or crossbred beef cows were anaesthetized on Days 15 to 17 after oestrus; the ovarian artery (OA), ovarian (utero-ovarian) vein (OV) and a peripheral artery (PA), were catheterized for chronic blood sampling. Beginning on the day after surgery, 6 sequential blood samples were collected every 30-40 min twice daily from 8 cyclic and 6 pregnant cows during Days 16-20: 818 blood samples (including 216 OA and PA concurrent arterial pairs) were collected. Overall least squares means for PGF concentrations (pg/ml) in the OV, OA and PA of cyclic cows were 562, 228 and 106, respectively. A significant (P less than 0.01) OA-PA difference (122 pg/ml) suggests that a local transfer system, between uterine venous effluent and ovarian arterial affluent, is functional in the cow. A transfer efficiency of about 1% was estimated. In cyclic cows differences in OA-PA concentrations of PGF were minimal on Days 16-18 and increased to about 160 pg/ml during luteal regression (Days 19-20). In pregnant cows a biphasic OA-PA pattern of difference in PGF between days was detected, with a peak on Day 18 (136 pg/ml) which was not apparent on Days 19-20. Amplitude of PGF spikes in the OA was significantly higher in cyclic (725 pg/ml) than in pregnant cows (397 pg/ml). We suggest that pregnancy suppresses PGF delivery to the ovarian circulation, resulting in maintenance of the corpus luteum in pregnant cows.