Showing papers by "Stephen J. O'Brien published in 1991"

Reconstruction of the chronically insufficient anterior cruciate ligament with the central third of the patellar ligament.

Abstract: The results of reconstruction of the anterior cruciate ligament with the central third of the patellar ligament as a free, autogenous, non-vascularized graft were retrospectively reviewed at our institution. Eighty reconstructions in seventy-nine patients were evaluated after a minimum of two years. In forty-eight (60 per cent) of the knees, the reconstruction was augmented with an extra-articular lateral sling of iliotibial band. The patients were evaluated with a physical examination, a KT-1000 arthrometer, radiographs, a subjective questionnaire, and a revision of the scale of The Hospital for Special Surgery for rating ligaments. Postoperatively, seventy-six (95 per cent) of the eighty knees no longer gave way, and the pivot-shift test was negative in sixty-seven (84 per cent) of the knees. The average score on the ligament-rating scale was 93 points. All of the patients who had clinical instability at the time of the most recent follow-up had associated ligamentous instability that had not been appreciated or addressed at the time of reconstruction. Arthrometric evaluation revealed that the laxity differed by three millimeters or less from that of the untreated knee in sixty (76 per cent) of the treated knees. In the patient who had bilateral reconstruction, the laxity was the same in both knees. Seventeen patients, who had more than three millimeters of translation, also had additional related ligamentous instability, most commonly posterolateral instability and insufficiency of the medial collateral ligament. We think that major associated ligamentous instability predisposes the reconstruction to failure and should be corrected in conjunction with the reconstruction.(ABSTRACT TRUNCATED AT 250 WORDS)

Bureaucratic Mischief: Recognizing Endangered Species and Subspecies

Abstract: of the Act by well-intentioned government officials. The listing of certain species as endangered has encouraged an increase in investigation of these taxa, notably in molecular genetics and field ecology (1). In some cases the molecular genetic results contradicted previous ideas about species integrity or taxonomic distinctions that were based on phenotypic (morphological) descriptions. Unfortunately these traditional taxonomic designations have been and continue to be the bases for management and eligibility for protection. This is a significant problem because the Endangered Species Act not only protects listed taxa from hunting, habitat exploitation, and other perils associated with human coexistence, but also provides significant financial resources for the effort to protect these species and to stabilize their populations. To illustrate the problem we summarize the interpretive difficulties posed by molecular results for four endangered groups. The Florida panther. This is a small population of mountain lion (also called cougar or puma) that descended from the Felis concolor coryi subspecies that ranged throughout the southern United States in the 19th century (2). The few remaining panthers (c50) living in southern Florida show significant physiological and reproductive impairments that are likely the consequence of inbreeding depression. A recent allozyme and mitochondrial DNA (mtDNA) analysis of the population revealed that two very distinct genetic stocks were living in Florida (2), one that resembled other North American pumas and another that was more closely related to a puma subspecies that had evolved in South America. Apparently seven animals from a captive stock (that later turned out to be a mixture of authentic F. concolor coryi and South American founders) were released into the Everglades between 1957 and 1967 and promptly forgotten. Today the founder ecosystem contains a mixture of two subspecies. The genetic advantages of introducing some additional genetic material into a population suffering from inbreeding would have been comforting except for one detail. Three opinions from the Solicitor's Office of the Department of the Interior (which is the counsel of the U.S. Fish and Wildlife Service) have ruled with the force of precedent that hybrids between endangered species, subspecies, or populations cannot be protected. Their opinions, referred to here as the Hybrid Policy, concluded that protection of hybrids would not serve to recover listed species and would likely jeopardize S.J. O'Brien is chief of the Laboratory of Viral Carcinogenesis, National Cancer

Case Study of a Population Bottleneck: Lions of the Ngorongoro Crater

Abstract: Lions in the Ngorongoro Crater, Tanzania, form a small and naturally isolated population. In 1962, the Crater lions suffered an epizootic that reduced the population to nine females and one male. An additional seven males apparently immigrated into the Crater in 1964–1965, but there has been no further immigration into the Crater in the past 25 years. By 1975, the population had recovered to its current level of 75-125 animals. All members of the current Crater population are descended from only 15 founders, and over the years there has been considerable variance in the reproductive success of both sexes. The Crater was probably colonized by lions from the nearby Serengeti ecosystem and the contemporary Crater lion population shows a significant lack of genetic diversity compared to the much larger Serengeti population. The detailed reproductive history of the Crater population was incorporated into a series of stochastic computer simulations that generated distributions of expected allele frequencies under different sets of initial conditions. The simulations suggest that the Crater population may have passed through previous bottlenecks before 1962 but that the level of heterozygosity in the breeding population has been declining since the mid-1970s, regardless of the population's genetic composition in the 1960s. High levels of inbreeding are correlated with increased levels of sperm abnormality in lions and there is evidence that the reproductive performance of the Crater lions has decreased as a result of decreasing heterozygosity.

Analytical DNA Fingerprinting in Lions: Parentage, Genetic Diversity, and Kinship

Abstract: The application of hypervariable minisatellite genomic families to the reconstruction of population genetic structure holds great promise in describing the demographic history and future prospects of free-ranging populations. This potential has not yet been realized due to unforeseen empirical constraints associated with the use of heterologous species probes, to theoretical limitations on the power of the procedure to track genic heterozygosity and kinship, and to the absence of extensive field studies to test genetic predictions. We combine here the technical development of feline-specific VNTR (variable number tandem repeat) families of genetic loci with the long-term demographic and behavioral observations of lion populations of the Serengeti ecosystem in East Africa. Minisatellite variation was used to quantify the extent of genetic variation in several populations that differed in their natural history and levels of inbreeding. Definitive parentage, both maternal and paternal, was assessed for 78 cubs born in 11 lion prides, permitting the assessment of precise genealogical relationships among some 200 lions. The extent of DNA restriction fragment sharing between lions was empirically calibrated with the coefficient of relatedness, r, in two different populations that had distinct demographic histories. The results suggest that reliable estimates of relative genetic diversity, of parentage, and of individual relatedness can be achieved in free-ranging populations, provided the minisatellite family is calibrated in established pedigrees for the species.

Molecular distance and divergence time in carnivores and primates.

Abstract: Numerous studies have used indices of genetic distance between species to reconstruct evolutionary relationships and to estimate divergence time. However, the empirical relationship between molecular-based indices of genetic divergence and divergence time based on the fossil record is poorly known. To date, the results of empirical studies conflict and are difficult to compare because they differ widely in their choice of taxa, genetic techniques, or methods for calibrating rates of molecular evolution. We use a single methodology to analyze the relationship of molecular distance and divergence time in 86 taxa (72 carnivores and 14 primates). These taxa have divergence times of 0.0 l-55 Myr and provide a graded series of phylogenetic divergences such that the shape of the curve relating genetic distance and divergence time is often well defined. The techniques used to obtain genetic distance estimates include one- and two-dimensional protein electrophoresis, DNA hybridization, and microcomplement fixation. Our results suggest that estimates of molecular distance and divergence time are highly correlated. However, rates of molecular evolution are not constant; rather, in general they decline with increasing divergence time in a linear fashion. The rate of decline may differ according to technique and taxa. Moreover, in some cases the variability in evolutionary rates changes with increasing divergence time such that the accuracy of nodes in a phylogenetic tree varies predictably with time.

The iliotibial band lateral sling procedure and its effect on the results of anterior cruciate ligament reconstruction

Abstract: The effect of iliotibial band "lateral sling" augmentation on long-term outcome in ACL reconstruction using free autogenous central-third patellar tendon was evaluated retrospectively. Eighty reconstructions were reviewed; the minimum followup was 2 years and the average was 4 years. Sixty percent of the procedures involved supplementation with a lateral sling. By both clinical evaluation and KT-1000 measurement, there were no differences in the results of patients with lateral sling augmentation and those without it. In addition, 40% of patients had chronic pain and/or swelling related to the lateral sling. We do not currently recommend the routine use of a lateral sling augmentation for ACL reconstruction using central-third patellar tendon.

Rescue and maturation in vitro of follicular oocytes collected from nondomestic felid species.

Abstract: The potential for rescuing immature oocytes from the ovaries of females of rare felid species which die or undergo medical ovariohysterectomy was evaluated. Ovaries were recovered from 13 species representing 35 individuals in good-to-poor health. Although the majority of females were 10 yr of age or older and in fair-to-poor health, a total of 846 oocytes were recovered of which 608 (71.9%) were classified as fair-to-excellent quality. One hundred of these oocytes were used for initial maturation classification and as parthogenetic controls. Overall, of the 508 fair-to-excellent quality oocytes placed in culture, 164 (32.3%) matured to metaphase II in vitro. For species in which 3 or more individuals yielded oocytes, mean oocyte maturation rates were as follows: 36.2%, tiger; 27.9% leopard; and 8.3%, cheetah. In vitro insemination of oocytes resulted in fertilization (2 polar bodies, 2 pronuclei, or cleavage) rates of 9.1% to 28.6% (leopard) using homologous fresh spermatozoa and 4.0% (lion) to 40.0% (puma) using homologous frozen-thawed spermatozoa. Inseminations using heterologous (domestic cat) spermatozoa also resulted in fertilized oocytes in the tiger, leopard, snow leopard, puma, serval, and Geoffroy's cat (range in fertilization rate, 5.0% for leopard to 46.2% for puma). Cleaved embryos resulted from the insemination of leopard oocytes with homologous sperm (n = 1 embryo) and puma oocytes with domestic cat sperm (n = 3 embryos). These results demonstrate that immature ovarian oocytes from rare felid species can be stimulated to mature in vitro despite an excision-to-culture interval as long as 36 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of Temperature and Gas Atmosphere on In-Vitro Fertilization and Embryo Development in Domestic Cats

Abstract: The influence of culture temperature and gas atmosphere on in-vitro fertilization and embryo development was examined in the domestic cat. In Exp. 1, eggs were fertilized and cultured in 5% CO2 in air at 37, 38 or 39 degrees C. Experiment 2 evaluated the effects of 5% CO2 in air; 5% CO2, 5% O2 and 90% N2; and 10% CO2 in air. Fertilization (cleavage) and development to the morula/blastocyst stage were not influenced (P greater than 0.05) by variations in temperature and gas composition. Despite changing these culture conditions, egg cleavage averaged approximately 75% and greater than 80% of the 2-cell embryos proceeded to morulae in vitro. However, the partial in-vitro morula-to-blastocyst developmental block normally observed in this species was not removed.

Culture medium and protein supplementation influence in vitro fertilization and embryo development in the domestic cat.

Abstract: The influence of culture medium and protein supplements on in vitro fertilization (IVF) and morula-to-blastocyst development in culture was examined in the domestic cat. In Study I, follicular oocytes were fertilized and cultured in 1) modified Krebs Ringer bicarbonate (mKRB); 2) modified Tyrode's solution (TALP) without phosphate or glucose; or 3) Ham's F10. All media contained bovine serum albumin (BSA). Fertilization rates were similar (P greater than .05) among mKRB (75.0%), TALP (70.6%), and Ham's F10 (80.0%) treatments. Compared to TALP (77.8%), more (P less than .05) embryos in Ham's F10 (95.0%) developed to the morula stage; development of mKRB embryos (88.9%) was intermediate and not different (P greater than .05). Study II evaluated the effects of protein availability and type on IVF and embryo growth. Ham's F10 was supplemented with polyvinylalcohol (PVA, 2 mg/ml), BSA (4 mg/ml), fetal calf serum (FCS, 5%), or estrous cat serum (ECS, 5%) and used to fertilize and culture embryos. Fertilization was enhanced (P less than .05) using either FCS (84.0%) or ECS (85.2%) compared to PVA (67.3%). Oocytes exposed to BSA fertilized at a rate (76.1%) similar (P greater than .05) to other treatments. Embryos exposed to all four treatment groups (PVA, 82.8%; BSA, 82.8%; FCS, 92.9%; or ECS, 97.8%) were equally capable (P greater than .05) of becoming morulae. However, more FCS- or ECS-supplemented morulae continued to the early blastocyst stage (30.8%, 22.2%, respectively, P less than .05) than PVA- (10.3%) or BSA- (13.8%) exposed morulae. In the domestic cat IVF system, the type of medium and protein supplement used appears to have a greater impact on embryo development in vitro than on fertilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Detecting high-resolution polymorphisms in human coding loci by combining PCR and single-strand conformation polymorphism (SSCP) analysis.

Abstract: A strategy is described that allows the development of polymorphic genetic markers to be characterized in individual genes. Segments of the 3' untranslated regions are amplified, and polymorphisms are detected by digestion with frequently cutting enzymes and with the detection of single-stranded conformation polymorphisms. This allows these genes, or DNA segments, to be placed on the linkage maps of human chromosomes. Polymorphisms in two genes have been identified using this approach. A HaeIII polymorphism was detected in the KIT proto-oncogene, physically assigned to chromosome 4q11-12. This polymorphism is linked to other chromosome 4p markers and is in linkage disequilibrium with a HindIII polymorphism previously described at this locus. We have also identified in the insulin-like growth factor1 receptor gene (IGF1R) a 2-bp deletion that is present at a frequency of .25 in the Caucasian population. Pedigree analysis with this insertion/deletion polymorphism placed the IGF1R gene at the end of the current linkage map of chromosome 15q, consistent with the physical assignment of 15q2526. Thus, polymorphisms in specific genes can be used to related the physical, genetic, and comparative maps of mammalian genomes and to simplify the testing of candidate genes for human diseases.

Genetic mapping of the beta 1 GABA receptor gene to human chromosome 4, using a tetranucleotide repeat polymorphism.

Abstract: As more coding loci for functional human genes are described, there is a growing need to identify DNA polymorphisms in specific genes. By examining DNA sequences within the introns of the beta 1 subunit of the gamma-aminobutyric acid receptor gene, GABARB1, we found a tetranucleotide repeat sequence (GATA). Amplification of this region by using PCR revealed seven alleles and a high degree of polymorphism (PIC = .75) in human populations. DNAs from the CEPH families were typed for the GABARB1 intron polymorphism and were analyzed with respect to 20 linked markers on chromosome 4. The results permit placement of GABARB1 on the linkage map of chromosome 4, between D4S104 and ALB. These results affirm that sequence analysis of noncoding segments included within or adjacent to functional genes has value as a strategy to detect highly informative polymorphisms.

Does tourniquet use during anterior cruciate ligament surgery interfere with postsurgical recovery of function? A review of the literature

Abstract: This article explores the issue of whether use of a tourniquet during arthroscopically assisted repair of the anterior cruciate ligament (ACL) slows the postoperative recovery of function. A tourniquet is customarily used to provide a bloodless field, thus enabling the surgeon to visualize the joint clearly. However, there is increasing evidence that tourniquets cause muscle and nerve damage that can have long-term consequences for the recovery of function following surgery. The two randomized trials that investigated tourniquet use during meniscectomy reached contradictory conclusions about the effects of the tourniquet. There have been no randomized trials of tourniquet use during the longer and more complex ACL surgery. This article reviews the pertinent literature and suggests some clinical implications of the available information.

Developmental changes in pituitary–gonadal function in free-ranging lions (Panthera leo leo) of the Serengeti Plains and Ngorongoro Crater

Abstract: activity. In contrast, there were no age-related differences in ejaculate characteristics of Ngorongoro Crater lions. Seminal quality in the Crater population was poor in adult and young adult animals and was unrelated to alterations in pituitary or testicular function. In summary, only seminal quality in adult male lions was affected by location, whereas age significantly affected both basal and GnRH-stimulated testosterone secretion and seminal quality (Serengeti Plains only) in sexually mature males. The striking seminal/ endocrine differences among pride (breeding) males of different ages raises questions about the impact of age on individual reproductive performance in this species.

Semen characteristics in free-living koalas (Phascolarctos cinereus)

Abstract: Spermic electroejaculates (range in motile sperm/ejaculate, 0.50-122.9 x 10(6); mean +/- s.e.m., 38.6 +/- 4.9) were recovered from 47 of 48 adult koalas captured from 3 wild populations in Australia. Semen was characterized by (i) a high density of globular bodies, which prevented the estimation of sperm motility without dilution; (ii) a brownish colour; and (iii) an acidic pH. Spermatozoa were categorized on the basis of 10 head forms, most cells being a curved or hooked shape. The koala populations differed in sperm concentration and motility ratings, but not in testes size, testosterone production or proportions of spermatozoa with various head shapes. These data confirm that free-living koalas normally produce spermatozoa with a high incidence of structural heterogeneity almost solely confined to the head region; and demonstrate the utility and safety of conventional gamete and endocrine studies, approaches which will be useful for determining the impact of genetic isolation and venereal disease on species fertility.

Genetic variation in Sri Lankan leopards

Abstract: Electrophoretic variation of 50 gene-enzyme systems was typed in a population of 33 captive leopards (Panthera pardus) from the island of Sri Lanka. The captive leopard population was composed of several lineages: (1) wild-caught leopards of the island subspecies (P. p. kotiya), (2) captive-born animals of the same subspecies, (3) a melanistic lineage whose founders were obtained from Malaysia (P. p. delacouri), and (4) leopards of known mixed lineage and unknown status. Two loci, APRT and PGD, were polymorphic in all samples, whereas 48 loci were invariant. Percent polymorphism (P) and percent average heterozygosity (H) were calculated as 4% and 1.4%, respectively, for the wild-caught leopards; 4% and 1.2% for the captive-born kotiya leopards; and 4% and 2.0% for the melanistic lineage. The overall results revealed a detectable decrease in genetic variability compared with a previous study of captive leopards from mainland origins. The mainland leopards had three additional polymorphic loci, ADA, ESI, and HBB. Reexamination of the TF locus using a revised protocol resolved a new allele in the sample of mainland leopards but not in the Sri Lankan sample. With this new polymorphism, recalculated P and H values for the mainland sample are 10% and 3.1%, respectively. No significant differences in polymorphic loci were observed between the leopard subspecies examined.

Linkage mapping of the human CSF2 and IL3 genes

Abstract: Interleukin 3 (encoded by the IL3 gene) and granulocyte-macrophage colony-stimulating factor (encoded by the CSF2 gene) are small secreted polypeptides that bind to specific cell surface receptors and regulate the growth, gene expression, and differentiation of many of the hematopoietic cell lineages, particularly nonlymphoid cells. The IL3 and CSF2 genes have been cloned and mapped to human chromosome bands 5q23-31. Only 10 kilobases of DNA separates the two genes, suggesting that they have a common origin and/or regulation. We have cloned 70 kilobases of genomic DNA that includes the IL3 and CSF2 genes, as well as flanking sequences, and report a physical map of this region. Several unique-sequence DNA segments have been identified in this region, and one of these fragments detects two restriction fragment length polymorphisms in DNA from unrelated Caucasians. Segregation of these DNA polymorphisms was followed in the Centre Etude du Polymorphisme Humaine (CEPH) panel of 40 large three-generation pedigrees, and linkage was detected with 17 genetic markers previously typed in these families. Multipoint linkage analysis permits the placement of the region containing the IL3 and CSF2 structural genes on the recombination-genetic linkage map of chromosome 5q and thereby allows the role of these genes in leukemogenesis to be more critically examined.