The outcome of patients in traumatic cardiac arrest presenting to deployed military medical treatment facilities: data from the UK Joint Theatre Trauma Registry.

BACKGROUND: The UK military was continuously engaged in armed conflict in Iraq and Afghanistan between 2003 and 2014, resulting in 629 UK fatalities. Traumatic cardiac arrest (TCA) is a precursor to traumatic death, but data on military outcomes are limited. In order to better inform military treatment protocols, the aim of this study was to define the epidemiology of TCA in the military population with a particular focus on survival rates and injury patterns. METHODS: A retrospective database analysis of the UK Joint Theatre Trauma Registry was undertaken. Patients who were transported to a UK deployed hospital between 2003 and 2014 and suffered TCA were included. Those patients injured by asphyxiation, electrocution, burns without other significant trauma and drowning were excluded. Data included mechanism of injury, Injury Severity Score (ISS), Abbreviated Injury Scale (AIS) for each body region and survival to deployed (Role 3) field hospital discharge. RESULTS: 424 TCA patients were identified during the study period; median age was 23 years, with a median ISS of 45. The most common mechanism of injury was explosive (55.7%), followed by gunshot wound (38.9%), road traffic collision (3.5%), crush (1.7%) and fall (0.2%). 45 patients (10.6% (95% CI 8.0% to 13.9%)) survived to deployed (Role 3) hospital discharge. The most prevalent body region with a severe to maximum AIS injury was the head, followed by the lower limbs, thorax and abdomen. Haemorrhage secondary to abdominal and lower limb injury was associated with survival; traumatic brain injury was associated with death. CONCLUSIONS: This study has shown that short-term survival from TCA in a military population is 10.6%. With appropriate and aggressive early management, although unlikely, survival is still potentially possible in military patients who suffer traumatic cardiac arrest.

Reproductive performance of ewes after 5-day treatment with intravaginal inserts containing progesterone in combination with injection of prostaglandin f2alpha.

Three experiments were conducted with a total of 1579 ewes to examine reproductive performance in response to synchronization of oestrus during the breeding season, using controlled internal drug releasing (CIDR-G) inserts in regimens designed to provide high concentrations of circulating progesterone. In experiment 1, treatment with two CIDR-G inserts for 12 days produced conception rate (79%) and prolificacy (1.9) to first service equivalent to breeding at natural oestrus (56% and 2.0, respectively). Pregnancy rates to two service periods were 90 and 79%, respectively. In experiments 2 and 3, progesterone was delivered by a single CIDR-G insert for 5 days in combination with prostaglandin F2alpha (PGF2alpha; 5 mg i.m., twice, 3 h apart) the day before (experiment 2), or at insert removal (experiment 3). The combined treatments improved rates of synchronization of oestrus (p<0.01) by 23 and 20% points, respectively, and pregnancy rates to the first service period by 19 (p<0.05) and 13 (p<0.01) percentage points, respectively, compared to treatment with PGF2alpha alone. It is concluded that the combination of treatment for 5 days with a CIDR-G insert and two injections of 5 mg PGF2alpha, the day before, or the day of insert removal, were effective treatments to obtain high fertility at synchronized oestrus in ewes during the breeding season.

Fertility response of yearling beef heifers after prebreeding energy manipulation, estrous synchronization and timed artificial insemination.

The effect of dietary energy and weight class on the fertility of yearling beef heifers (Angus, Hereford, and Angus x Hereford) was investigated over 2 years. In year 1, heifers (n=58) were classed as heavy (HW; > or =340 kg) or light weight (LW; <340kg) and then assigned to receive either a low (LE; 0.23 kg/day) or high energy (HE; 0.68 kg/day) diet. In year 2, heifers (n=60) were also classified as heavy (> or =335kg) or light weight (<335 kg), but the energy content of the diet was raised so that heifers on the LE and HE were targeted to gain 0.46 and 0.79 kg/day, respectively. Heifers in the four groups, LELW (n=14 and 12), LEHW (n=16 and 17), HELW (n=13 and 15), and HEHW (n=15 and 16) received restricted amounts of concentrate (HE > LE) and free choice hay over 47 or 42 days (year 1 and year 2, respectively). To synchronize estrus, heifers were fed capsules containing MGA (0.5 mg/animal each day) beginning 11 days before the end of the feeding trial (day 0), PGF(2alpha) (25mg i.m.) and estradiol benzoate (Ebeta; 400 microg i.m.) was given on days 8 and 10, respectively. Estrous behavior was observed (days 10 and 11) and all heifers were inseminated on day 11. Following AI, heifers were re-grouped and a bull was introduced (days 27-39) for the second service in both years. Pregnancy diagnosis for the first (days 41-42) and second services (days 69-97) was performed by transrectal ultrasonography. Transrectal ultrasonic observations of ovarian follicle number and size were completed for a subset of heifers (n=5-8) from each experimental group at the end of the feeding trial. The effect of year was not significant for any of the reproductive performance variables measured. The mean ADG was (0.72 +/- 0.04 kg/day) and was greater in LW than HW heifers and in heifers in the HE than LE treatment groups (P <0.05). In heifers receiving the LE diet, ADG was lower in HW than LW heifers (weight x diet; P=0.02; 0.54 +/- 0.04 and 0.62 +/- 0.03 kg/day for HW and LW heifers, respectively). The diameter of the largest follicle was greater in heifers receiving the HE diet (P < 0.05; 11.3 +/- 0.4 mm) than those on the LE diet (10.3 +/- 0.3), and in LW (P <0.05) compared to HW heifers. The HE diet increased the size of the largest follicle in LW but not HW heifers (diet x weight, P <0.05). The percentage of pubertal heifers at the end of the feeding period (59.3%), estrous response (56.4%), conception rate (47.7%), ovulation rate (88.9%), and first service pregnancy rate (36.2%) were not significantly affected by initial weight or diet. There was a tendency for first service pregnancy rates to be greater in LW than HW heifers consuming the LE diet (diet x weight, P <0.1; 54.2 +/- 15 and 30.3 +/- 10% for LELW and LEHW heifers, respectively). Pregnancy rate after two services was greater (P=0.01) in LW (82 +/- 10%) than in HW (64.5 +/- 10%). The LE diet achieved moderate rates of gain and allowed high level of reproductive performance in LW but not HW heifers.

Ability of ram introduction to induce LH secretion, estrus and ovulation in fall-born ewe lambs during anestrus.

The ability of ram introduction (RI) and progesterone pre-treatment to induce increases in LH secretion and ovulation, and the ability of progesterone pre-treatment with or without estrogen to induce estrus and ovulation in fall-born ewe lambs during seasonal anestrus was investigated. In early July, lambs of mixed breeds (41.8+/-0.6 kg and 250.7+/-1.3 days of age) were assigned to receive no treatment (C, n=7), to be introduced to rams (7:1 ewe:ram ratio; R, n=7), to be treated with progesterone (a used CIDR device) for 5 days (P, n=5), to be treated with progesterone and introduced to rams at CIDR removal (PR, n=11), or to receive the latter treatment plus an injection of estradiol benzoate (25 microg, E2beta i.m.) 24 h after CIDR withdrawal/RI (PER, n=11). Blood samples were collected from all lambs every 4h for 60 h beginning at RI/CIDR withdrawal (0 h), to characterize the LH surge profile and in groups R and C every 15 min for 8 h between 12 and 20 h for determination of LH pulse frequencies. Ultrasonographic examinations of the ovaries were conducted at 0, 36 and 60 h. In ram-exposed groups lambs were also observed for raddle marks every 4h from 0 to 60 h. The LH pulse frequency (pulses/8 h) was higher in group R (P<0.01; 7.7+/- 0.5) than group C lambs (2.7+/- 0.8). More lambs in groups exposed to rams than in the C or P groups showed an LH surge (P<0.05; 0, 100, 0, 72.7 and 100%, for C, R, P, PR and PER groups, respectively). Time from RI/CIDR removal to initiation of the LH surge was greater in lambs in the PR (43.5+/- 3.8h) than in the R (32.6+/- 4.6h; P=0.08) or PER (33+/- 1.2h; P<0.01). Diameter of the largest follicle at 0 h (3.2+/- 0.2mm) was not different among groups. Growth rate of the largest follicle between 0 and 36 h was greater (P<0.05) in RI than in C or P groups. Diameter of the largest follicle at 36 h was larger (P<0.05) in lambs in R (5.6+/- 0.2mm) and PR (5.1+/- 0.5mm) than C (4.0+/- 0.6mm) or P (3.8+/- 0.4mm) groups, and in R than PER (4.3+/- 0.4mm) treatment groups. Only lambs in the RI groups ovulated. Among RI groups the percentage of lambs ovulating was greater in the R (P<0.05; 85.7%) than PR (33.3%) groups with an intermediate response observed in lambs in treatment group PER (71.4%). The estrous response in progesterone pre-treated groups was greater (P<0.05) in lambs also treated with estrogen (PER; 81.8%), than in lambs introduced to rams alone (PR; 45.5%). In conclusion, ram introduction by itself, but not progesterone treatment alone, induces increases in LH pulse frequency, follicular development, and ovulation in fall-born ewe lambs during seasonal anestrus, further, P4 pre-treatment and RI when combined with E2 results in a high estrous response.

Effectiveness of intravaginal progesterone inserts and FSH for inducing synchronized estrus and increasing lambing rate in anestrous ewes.

The objectives of this study were to determine whether a new progesterone (P4)-releasing intravaginal insert would induce fertile estrus and whether FSH combined with the insert would increase prolificacy in anestrous ewes introduced to rams. Ewes of mixed breeding on six farms were assigned to four randomized treatments: control (C), n = 73; 12 d P4 (polycapralactone [PCL] insert with 0.82 g P4), (P12), n = 73; 12 d P4 plus i.m. FSH (Folltropin, 55 mg NIH-FSH-P1 equivalent) in propylene glycol, 24 h before insert removal, (P12F), n = 71; and 5 d P4 plus FSH (P5F), n = 77. Growth and ovulation of follicles were observed ultrasonographically in 20 ewes at four farms (five/treatment) at insert removal and 36, 48, 72, and 96 h later. Intact rams (1:15 ewes in multiple-sire groups) were joined at insert removal, and raddle marks were observed every 12 h for 5 d. On d 26 to 30, rams were removed; ewes were examined for pregnancy then and 20 d later. Percentage of ewes marked by rams was greater in P4-treated (66 to 79%) than in C (12%; P < 0.01) ewes and in P5F (79%) than in P12F (66%; P < 0.05). Diameters of largest follicles at insert removal were greater (P < 0.05) in P4-treated (5.5 +/- 0.2) than in C ewes (4.8 +/- 0.2). Progesterone increased numbers of follicles > 3 mm (P < 0.01) or ovulated (P < 0.05; 2.6 +/- 0.6 vs 1.3 +/- 0.6 in C ewes) and FSH increased number of follicles > 3 mm (P < 0.05). In FSH-treated ewes, ovulation rate tended to be greater after treatment with P4 for 5 than for 12 d (P = 0.09, 3.3 +/- 0.6 and 2.2 +/- 0.4, respectively). More P4-treated than C ewes lambed (P < 0.01) to the first (38 to 45 vs 0%) or both (63 to 66 vs 41%) service periods. Prolificacy (first service) did not differ between FSH-treated ewes (P12F + P5F; 1.8 +/- 0.1) and ewes treated with P4 only (P12; 1.6 +/- 0.1). However, FSH increased prolificacy to first service (1.8 +/- 0.1) over prolificacy to second service (C ewes 1.5 +/- 0.1; P < 0.05, and all ewes 1.4 +/- 0.1; P < 0.01). Pregnancy retention did not differ among treatments but was greater (P < 0.01) in ewes that conceived at the first (90.9 +/- 3.7) than at the second (72.5 +/- 3.3) service period. In conclusion, a PCL insert in combination with ram introduction at insert removal was more effective than ram introduction alone to induce synchronized estrus and ovulation and to yield pregnancy after one or two service periods. Treatment with P4 for 5 d was as effective as for 12 d to induce fertile estrus in FSH-treated anestrous ewes.

Short-term treatment with a controlled internal drug releasing (CIDR) device and FSH to induce fertile estrus and increase prolificacy in anestrous ewes.

The objectives were to evaluate, in anestrous ewes, the effectiveness of a CIDR-G device (0.3 g progesterone) administered for 5 d to induce estrus; and FSH (Folltropin; 55 mg NIH-FSH-P1 equivalent) in saline:propylene glycol (1:4) 24 h before insert removal (Day 0), to increase ovulation rate and prolificacy. Ewes of mixed breeding were assigned at random to 3 treatments: control (C; n = 125), 5 d progesterone (P5; n = 257) and 5 d progesterone plus FSH (P5F; n = 271). Intact rams were joined at insert removal and ewes were observed every 24 h for 3 d. On Day 14, the ovulation rates of all ewes detected in estrus in the treated groups were determined using transrectal ultrasonography. Rams were removed on Day 26 to 31. Ewes were examined for pregnancy then, and again 20 to 25 d later to detect ewes that conceived to the second service period. Percentage of ewes marked by rams was higher in progesterone-treated (77%) than in C (20%; P < 0.01), but did not differ between P5 and P5F. The ovulation rate (1.95+/-0.04) did not differ due to FSH. Conception (68%) and pregnancy (52%) rates were higher in progesterone-treated (P < 0.01) than in C (0%) ewes. Estrous response varied quadratically with time after ram introduction, and the conception rate varied quadratically with the time of observation of onset of estrus. Over two service periods more progesterone-treated than C ewes lambed (65 vs 45%; P < 0.01). Lambs born per ewe exposed (0.7+/-0.1, 1.0+/-0.1, and 1.1+/-0.1 for C, P5 and P5F, respectively) was increased by progesterone (P < 0.05). Litter size to the first service period (1.59+/-0.04) and overall (1.54+/-0.03) did not differ among treatment groups. FSH-treated ewes tended to have more lambs (1.67+/-0.1) than did ewes receiving progesterone alone (1.5+/-0.1; P = 0.06) and than did ewes lambing to the second service period (1.5+/-0.1; P = 0.06). In summary, a 5-d progesterone pre-treatment of anestrous ewes induced estrous cycles and increased the pregnancy rates. A single injection of FSH only tended to increase litter size.

Ability of induced corpora lutea to maintain pregnancy in beef cows.

Experiments were conducted in beef cows without a primary CL, in which pregnancy had been maintained with exogenous progestogen. In preliminary trials, replacement CL induced ipsilateral to the embryo and after, rather than before, d 36 of pregnancy, maintained more pregnancies after withdrawal of exogenous progestogen (13/13 vs 2/6; P < 0.05). In Exp. 1, in cows with replacement CL induced by treatment with hCG on d 28 of pregnancy, treatment with flunixin meglumine on d 31 through 37 did not increase maintenance of pregnancy. Experiment 2 was conducted to evaluate directly the effects of concentrations of PGF2alpha and estradiol-17beta during d 31 through 35 of pregnancy on maintenance of pregnancy by replacement CL induced between d 28 and 31. In cows that maintained pregnancy while progestogen was provided, maintenance of pregnancy after withdrawal of exogenous progestogen tended to be greater with high (5/5) than with low (2/6; P < 0.10) concentrations of PGF2alpha and greater with low (6/7) than with high (2/6; P = 0.10) concentrations of estradiol-17beta. Secretion of progesterone by replacement CL was greater (P < 0.05) in cows with high than in those with low concentrations of PGF2, during d 31 through 35. Prostaglandin F2alpha may facilitate attachment of the bovine embryo (d 30 to 40) in a manner similar to that reported for implantation in other species. Cows that did not form CL in response to hCG on d 28 to 31 responded well when retreated after d 36. Again, maintenance of pregnancy was greater when replacement CL were induced after (9/9) rather than before d 36 (8/16; P < 0.05).

Effect of the previously gravid uterine horn and postpartum interval on follicular diameter and conception rate in beef cows treated with estradiol benzoate and progesterone.

An experiment was performed to evaluate the effect of the side of ovulation with respect to the previously gravid uterine horn on fertility of cows inseminated at one of two periods postpartum. All cows were treated with an intravaginal progesterone insert for 7 d and received estradiol benzoate (2 mg, i.m.) at the time of device insertion, prostaglandin F2alpha (25 mg, i.m.) at the time of device removal, and estradiol benzoate (1 mg, i.m.) 30 h after device removal. All cows were inseminated 28 to 30 h after the second treatment with estradiol benzoate, regardless of observed estrus. Cows treated in Period 1 received inserts at 16 to 20 d postpartum and were inseminated at 25 to 29 d postpartum. Cows treated in Period 2 received inserts at 26 to 30 d postpartum and were inseminated at 35 to 39 d postpartum. Diameter of the largest follicle at insert removal was greater in cows treated in Period 2 (10.1 +/- 0.3; mm +/- SEM) than in cows treated in Period 1 (9.1 +/- 0.3; P < .05). Diameter did not differ with the side of ovulation in respect to the previously gravid uterine horn. Diameter was greater in cows 5 to 9 (10.3 +/- 0.3) than in cows 3 to 4 (9.0 +/- 0.3) or 10 to 13 (9.4 +/- 0.6) yr of age (P < .01). The proportion of cows that ovulated from the ovary contralateral to the previously gravid uterine horn was greater (P < .05) than of those that ovulated from the ipsilateral ovary, and the incidence of ovulation was reduced in cows 3 to 4 yr of age (P < .01). Conception rate tended to be greater for ovulation from the ipsilateral compared with the contralateral ovary, relative to the previously gravid uterine horn (P < .10) and for ovulation from the right than the left ovary (P < .06). Conception rate was less if cows ovulated a follicle that was < 9 mm than a follicle > or = 9 mm in diameter at insert removal (P < .01) and was greater in cows inseminated in June than in April or May (P < .05). In conclusion, in cows in which estrus was synchronized at 25 to 39 d postpartum, ovulation from either the ovary ipsilateral to the previously gravid uterine horn, or the right ovary, tended to increase fertility.

Effects of long-term estrogen implants in beef heifers.

Crossbred beef heifers (n = 78) were assigned randomly to one of three treatments. Heifers received either no implant, one estradiol-releasing implant (Compudose), or two estradiol-releasing implants. Heifers were implanted at birth and then reimplanted every 150 d. Calves were maintained with the cows until weaning at approximately 200 d of age. Heifers were placed in the feedlot as one group and fed a growing diet for 56 d. Following the growing phase, heifers were segregated into their respective treatment groups and fed until selected by industry buyers for harvest. Beginning at 1 yr of age and continuing every 14 d until puberty or harvest, heifers were palpated per rectum, and blood samples were collected for determination of ovarian activity and attainment of puberty. Serum progesterone of > or =1 ng/mL and(or) palpation of a detectable corpus luteum were criteria of puberty. At weaning and again at harvest, an x-ray was taken of the left front leg of six heifers selected randomly from each group. Metacarpal bones III and IV from the same animals were collected at harvest and transected for determination of epiphyseal plate closure. The x-ray scores and the actual measurements had a correlation of .94. Epiphyseal plate closure occurred in a dose-related manner, with heifers on the higher dose of estrogen having earlier plate closure than heifers on the lower dose. At harvest, reproductive tissues and carcass data were collected for all heifers. Eighteen of 25 untreated control heifers (P < .05), 6 of 26 heifers treated with one implant, and 2 of 26 heifers treated with two implants attained puberty by the end of the experiment. No differences (P > .10) were detected among treatment groups for carcass traits. These data suggest that early and continuous treatment of heifers with estradiol implants can retard reproductive function.

Chromosomal analysis of renal angiomyolipoma by comparative genomic hybridization: evidence for clonal origin.

Angiomyolipoma has long been considered a hamartomatous polyclonal proliferation. However, recent molecular analyses have indicated that these tumors may be clonal neoplasms rather than polyclonal proliferations. We investigated chromosomal imbalances in angiomyolipoma by comparative genomic hybridization. DNA was extracted from archival paraffin-embedded and frozen tissues of 12 angiomyolipomas (10 usual variant, two epithelioid variant). The 10 angiomyolipomas of the usual variant included bilateral tumors from one tuberous sclerosis patient. Fluorescence ratio distributions from tumor hybridizations were compared with those from control hybridizations to detect changes in DNA copy number with high sensitivity and specificity. We identified 20 chromosomal imbalances in seven sporadic angiomyolipomas, including both tumors of the epithelioid variant. The remaining five tumors, including the two angiomyolipomas from a tuberous sclerosis patient, were devoid of chromosomal imbalances. Seventy-five percent of the imbalances were partial or whole chromosomal deletions involving disparate genomic regions, some of which have previously been associated with tumors of adipose tissue and smooth muscle tumors. Four angiomyolipomas of the usual variant showed 5q deletions with a common region of deletion spanning 5q33 to q34. In two tumors, deletion on 5q was the sole abnormality. One epithelioid angiomyolipoma showed 5q gain encompassing the same region in addition to other alterations. We concluded that (1) Chromosomal imbalances are common in renal angiomyolipomas; (2) Presence of clonal genomic alterations lends further support to the neoplastic pathogenesis of these tumors; (3) The 5q33-q34 region may contain a tumor suppressor gene significant in the histogenesis of some renal angiomyolipomas.

Follicular growth, estrus and pregnancy after fixed-time insemination in beef cows treated with intravaginal progesterone inserts and estradiol benzoate.

An experiment was performed to compare the effects of 3 short-term treatments with progesterone and estradiol benzoate (EB) on follicular growth, synchrony of estrus and pregnancy rate after fixed-time insemination in lactating postpartum beef cows. In Treatment 1 (n = 46), each cow received a progesterone-containing intravaginal insert for 7 d with injection of EB (2 mg, i.m.) at the time of device insertion. In Treatment 2 (n = 46), the insert was used for only 5 d with injection of EB (2 mg, i.m.) at the time of insertion. Cows in Treatment 3 (n = 47) received an insert for 5 d with no EB at the time of insertion. Each cow in the 3 groups received PGF2 alpha (25 mg, i.m.) at the time of insert removal, followed by EB (1 mg, i.m.) 30 h later. The cows were then inseminated 28 to 30 h after treatment with EB (58 to 60 h after insert removal). Treatment with 2 mg EB terminated the growth of the largest ovarian follicle (> 5 mm in diameter) at device insertion in 16/16 and 14/15 cows in Treatments 1 and 2, respectively. Estrus was detected within an 8-h target period (48 to 56 h after insert removal) in 93, 87 and 81% of cows in Treatments 1, 2 and 3, respectively (P > 0.05). Pregnancy rates at 39 d post insemination were 60, 50 and 51% for Treatments 1, 2 and 3, respectively (P > 0.05). The pregnancy rates did not differ between cows that were anovulatory or those that had ovulated before the initiation of treatments (54%), or among cows that were 28 to 40, 41 to 60 or > 60 days post partum at insemination (43, 59 and 54%, respectively). Treatment with progesterone inserts for 5 or 7 d, PGF2 alpha at the time of insert removal and 1 mg EB 30 h later induced the high degree of synchrony of estrus and ovulation necessary for fixed-time insemination.

A similar pattern of chromosomal alterations in prostate cancers from African-Americans and Caucasian Americans.

A combination of genetic and epigenetic factors may explain the disproportionate incidence and mortality of prostate cancer among African-American males (AAMs) as compared with Caucasian American males (CAMs). We wished to determine whether primary prostate cancers from AAMs and CAMs harbor different patterns or frequencies of chromosomal alterations. Comparative genomic hybridization (CGH) was performed on clinically localized, untreated primary prostate cancers from 16 AAMs and 16 CAMs. Detailed statistical analysis was used to delineate gains and deletions with high sensitivity and specificity and to compare the frequency and pattern of alterations between the two groups of tumors. The two groups of patients had indistinguishable preoperative serum prostate-specific antigen levels, and the two groups of tumors had similar pathological stages and grades. Chromosomal gains and deletions occurred in regions known to be frequently altered in prostate cancer. Specifically, the most frequent alterations were deletions of regions on chromosomes 13q, 5q, 16q, and 8p and gains of regions on 8q and 5q. When tumors from AAMs and CAMs were compared, the frequencies of alteration (deletion, gain, or no alteration) were similar across 98.9% of the length of the genome. The patterns of alterations of the most frequently altered chromosomes were also similar between tumors from AAMs and CAMs. We concluded that primary prostate cancers from AAMs and CAMs harbor a similar pattern and frequency of chromosomal alterations. These data support the notion that sporadic prostate cancers from AAMs and CAMs develop by similar chromosomal mechanisms. Biological differences, if present, do not occur on the chromosomal level.

Estrus and luteal function in suckled beef cows that were anestrous when treated with an intravaginal device containing progesterone with or without a subsequent injection of estradiol benzoate.

The objectives in this study were to determine whether treatment with progesterone (P4) via an intravaginal device would induce estrus and development of corpora lutea (CL) with typical life spans and whether treatment with estradiol benzoate (EB) following device removal would enhance the responses. At treatment initiation (d 0), suckled beef cows (n = 362) that were anestrous received one of the following: 1) an intravaginal device containing P4 for 7 d plus an injection of 1 mg of EB 24 to 30 h after device removal (P4+EB), 2) an intravaginal device containing P4 for 7 d (P4), 3) a sham device for 7 d plus an injection of 1 mg of EB 24 to 30 h after device removal (EB), or 4) a sham device for 7 d (control). Progesterone treatment increased the proportion of cows that formed CL with a typical lifespan (P < .001) and exhibited behavioral estrus (P < .05). Treatment with EB in combination with P4 increased (P < .001) the proportion of cows that exhibited estrus. Treatment with P4 alone had no effect, but the combination of P4 and EB increased (P < .01) the proportion of cows that formed short-lived or typical lifespan CL by the end of the experiment compared with untreated cows. In summary, treatment with P4 increased the incidence of estrus and the proportion of cows that formed CL with a typical lifespan. Treatment with P4 resulted in resumption of luteal function in suckled beef cows that were anestrous, with most cows developing CL with a typical lifespan, whereas EB enhanced the expression of estrus.

Novel and complex chromosomal arrangement of Rhizobium loti nodulation genes.

A mutational and structural analysis of Rhizobium loti nodulation genes in strains NZP2037 and NZP2213 was carried out. Unlike the case with other Rhizobium strains examined to date, nodB was found on an operon separate from nodACIJ. Sequence analysis of the nodACIJ and nodB operon regions confirm that R. loti common nod genes have a gene organization different from that of other Rhizobium spp. At least 4 copies of nodD-like sequences were identified in R. loti. The complete nucleotide sequence of one of these, nodD3, was determined. A new host-specific nod gene, nolL, was identified adjacent to nodD3. NolL shares homology with NodX and other O-acetyl transferases. Mutational analysis of the nod regions of strains NZP2037 and NZP2213 showed that nodD3, nodI, nodJ, and nolL were all essential for R. loti strains to effectively nodulate the extended host Lotus pedunculatus, but were not necessary for effective nodulation of the less restrictive host, Lotus corniculatus. Both nodD3 and nolL were essential for R. loti strains to nodulate Leucaena leucocephala.

Embryotoxicity of a regressing corpus luteum in beef cows supplemented with progestogen.

Postpartum cows with short-lived corpora lutea produce embryos that arrive at the uterus, but pregnancy rates are low even with exogenous progestogen. Four experiments were conducted to determine whether prostaglandin (PG) F2 alpha, known to cause early luteolysis, could have a direct effect on embryonic loss. Exogenous progestogen was injected s.c. twice daily in each experiment, starting 3 or 4 days after estrus (day of estrus = Day 0). Nonlactating, cycling beef cows were mated and injected i.m. every 8 h on Days 4 through 7 (experiment 1) or 5 through 8 (experiment 3) with either 15 mg PGF2 alpha or 3 ml saline. In experiment 1, cows in a third group received 1 g flunixin meglumine i.m. every 8 h. Ten of 18 PGF2 alpha-treated cows in experiment 3 were luteectomized on Day 5. Pregnancy rates were higher (p < 0.05) in cows given saline or flunixin meglumine (5 of 7) than in cows given PGF2 alpha (1 of 5) in experiment 1, and in cows given saline (6 of 9) or given PGF2 alpha, and luteectomized (8 of 10) than in cows given PGF2 alpha (2 of 8) in experiment 3. Postpartum beef cows, mated at weaning-induced first estrus, received i.m. injections every 8 h on Days 4 through 9 of 3 ml saline or 1 g flunixin meglumine (experiment 2); 14 flunixin meglumine-treated cows were luteectomized on Day 7. Pregnancy rates were higher in cows given flunixin meglumine and luteectomized (7 of 14) than in cows given saline (4 of 15) or flunixin meglumine alone (3 of 15; p < 0.05). In experiment 4, postpartum cows were luteectomized or sham-operated on Day 5. Pregnancy rates (2 of 13 and 2 of 14, respectively) did not differ. Thus, both reduction of endogenous PGF2 alpha and luteectomy were required for embryo survival in postpartum cows with short-lived corpora lutea, whereas luteectomy alone prevented effects of exogenous PGF2 alpha in cycling cows.

Effect of peripheral concentrations of progesterone on follicular growth and fertility in ewes.

The effects of progesterone (P4) on follicular growth and fertility in ewes were examined. In Experiment 1, 22 ewes received either one or three packets of P4 (5 g/packet) or an empty packet subcutaneously (sc) from Days 5 to 15 of the estrous cycle (estrus = Day 0). On Day 6, P4-treated ewes received 12.5 mg of prostaglandin F2 alpha. Follicles > or = 3 mm in diameter were observed via transrectal ultrasonography daily from Day 4 through estrus, corpora lutea (CL) were observed 5 to 7 d after estrus. Ewes with low (LOW; < or = 1 ng/ml; n = 5), intermediate (MED; >1 and <2 ng/ml; n = 10), or normal (NOR; > or =2 ng/ml; n = 7) P4 in jugular plasma on Days 7 through 15 differed in follicular development. The largest follicle at estrus was larger in ewes with LOW vs. MED and NOR P4 (7.8 +/- 0.3 vs. 6.9 +/- 0.2 mm; P < 0.05). Treatments differed in proportions of multiple-ovulating ewes, in which the oldest ovulatory follicle was first observed before Day 10 (LOW: 3 of 3, MED: 6 of 10, NOR: 0 of 5, respectively; P < 0.05). Estradiol was higher early in the treatment period in LOW ewes than in MED and NOR ewes (day x treatment; P < 0.05). In Experiment 2, ewes received 5 mg of P4 in corn oil (low progesterone [LP]; n = 51) or 2 ml of corn oil (CON; n = 49) sc every 12 hr on Days 6 through 14 of the estrous cycle before mating. LP ewes received 15 mg of prostaglandin F2 alpha on Day 6. Mean serum P4 on Days 7 through 15 was 0.6 +/- 0.1 ng/ml in LP and 1.9 +/- 0.1 ng/ml in CON ewes. Eleven LP and 12 CON ewes were scanned daily from Day 4 through mating, and in all ewes (n = 93), CL were counted 10 d after mating and embryos were counted at 25, 40, and 60 d of gestation. In multiple-ovulating ewes, day of cycle of appearance was earlier for the oldest (Day 6.1 +/- 0.8 vs. 10.4 +/- 0.8) but not second oldest (Day 11.7 +/- 1.0 vs. 12.2 +/- 0.9) ovulatory follicles in LP compared with CON ewes. The conception rate was lower in LP (72%) than in CON ewes (98%; P < 0.01). However, numbers of CL 10 d after mating, and in pregnant ewes, numbers of embryos 25 d after mating and lambs born, did not differ with treatment. In summary, low P4 increased the size of the largest follicles and the age of the oldest ovulatory follicles. Embryos resulting from the ovulation of older and younger follicles in the same ewe did not differ in their ability to survive.

Cloning of a DNA repeat element from horse: DNA sequence and chromosomal localization.

A DNA repeat element, revealed initially by digestion of horse DNA with TaqI, was cloned and characterized by Southern and in situ hybridization studies and nucleotide sequencing. The clone, e4/1, consisted of 32 tandem reiteration of a unit repeat of 21-22 bp, and produced multilocus DNA fingerprinting profiles that were useful for parentage analysis in horses. The tandem repeat element was shown by in situ hybridization to be localized in the centromeres of the acrocentric but not metacentric classes of horse chromosomes.

Families of tandemly repeated DNA elements from horse: cloning, nucleotide sequence, and organization.

DNA repeats, revealed initially by digestion of horse DNA with restriction enzymes, were cloned and characterized by cross-hybridization studies and nucleotide sequencing. The Sau-like family of tandem repeats contained two classes of repetitive elements with unit repeats of about 80 bp that shared no sequence similarity. Both unit repeats were present, frequently in tandem, in cloned segments of horse DNA of less than 600 bp. Evidence is presented, based on their ladderlike patterns of hybridization to horse DNA and their high level of similarity to published sequences of satellites from equine DNA, suggesting that they are centromerically located in the horse genome. The Sau-like tandem repeat families were specific to Equidae. Another class of repeats, GATA-like elements, which did not appear to have a centromeric distribution, was also cloned and sequenced.

Thirteen loci physically assigned to sheep chromosome 2 by cell hybrid analysis and in situ hybridization.

Sheep x hamster cell hybrids containing sheep metacentric Chromosome (Chr) 2 were produced by fusing blood leukocytes from normal sheep with hamster auxotrophic Ade F-minus mutants. Cell clones that were isocitrate dehydrogenase 1 (IDH1) positive were cytogenetically characterized, confirming that they contained sheep Chr 2. The following loci were newly assigned by Southern hybridization to sheep Chr 2: lipoprotein lipase (LPL), glycoprotein-4-beta galactosyltransferase 2 (GGTB2), neurofilament light polypeptide (68 kDa; NEFL), surfactant-associated protein 2 (SFTP2), lymphocyte-specific protein tyrosine kinase (LCK), and nebulin (NEB). These new assignments and the in situ localization of gelsolin (GSN) to sheep Chr 2pter-p24 are consistent with the predicted homology of cattle Chr 8 (U18) with sheep Chr 2p, and of cattle Chr 2 (U17) with sheep 2q. In addition, the assignment by cell hybrid analysis of loci previously mapped to Chr 2 in sheep, viz., cholinergic receptor, nicotinic, delta polypeptide (CHRND), collagen type III alpha 1 (COL3A1), fibronectin 1 (FN1), isocitrate dehydrogenase (IDH1), and villin 1 (VIL1), confirmed the localization of sheep syntenic group U11 to this chromosome. By nutritional selection and complementation of the hamster auxotrophic Ade F mutation, the multifunctional enzyme locus phosphoribosylaminoimidazolecarboxamide formyltransferase (AICAR transformylase)/IMP cyclohydrolase (inosinicase) (provisionally given the symbol PRACFT) has also been newly assigned to sheep Chr 2. This report significantly extends the number of loci physically mapped to sheep Chr 2 and confirms its close homology with cattle Chrs 2 and 8.