A major locus influencing plasma high-density lipoprotein cholesterol levels in the San Antonio Family Heart Study. Segregation and linkage analyses.

To detect and measure the effects of a single locus on quantitative variation in plasma concentrations of HDL cholesterol (HDL-C), we conducted statistical genetic analyses on data from 526 Mexican American individuals in 25 randomly ascertained pedigrees. By using maximum-likelihood complex segregation analysis, we found evidence for a major locus with a codominant mixture model that included the phenotypic means, standard deviations, relative frequency of a low HDL-C allele, and heritability for plasma HDL-C levels, plus the effects of sex (genotype specific), age-by-sex, age2-by-sex, plasma concentrations of apolipoprotein (apo)AI and triglycerides (genotype specific), exogenous sex hormone use, and menopausal status under an unrestricted general model. Inclusion of the four covariates (in addition to the sex and age-by-sex effects) accounted for nearly 79% of the variance in total plasma HDL-C levels.

Humoral immune response in a marsupial Monodelphis domestica: anti-isotypic and anti-idiotypic responses detected by species-specific monoclonal anti-immunoglobulin reagents.

We examined the humoral immune response of the laboratory opossum (Monodelphis domestica) as a model marsupial species. To evaluate antibody responses, IgM and IgG preparations were purified from the sera of naïve Monodelphis. These two immunoglobulin (Ig) preparations were used to generate specific murine monoclonal antibodies for use in ELISA-based serology.

Lipid and lipoprotein factors associated with variation in Lp(a) density.

To determine the facets of lipoprotein metabolism associated with variation in lipoprotein(a) [Lp(a)] density, we examined by density gradient ultracentrifugation 246 plasma samples exhibiting single apo(a) isoform band phenotypes. Estimated molecular weights of apo(a) isoforms ranged from 488 to 874 kD, and they accounted for approximately 80% of variation in Lp(a) density. After adjustment for variation in the protein composition, we found in univariate analyses that variation in residual Lp(a) density was associated with 10 different measures of lipoprotein size and concentration.

Developmental progression of Gpd expression from the inactive X chromosome of the Virginia opossum.

Metatherian (marsupial) mammals possess a non-random form of X-chromosome inactivation in which the paternally-derived X is always the one inactivated. To examine the progression of X-linked gene expression during metatherian development, we compared relative levels of the maternally and paternally encoded Gpd gene products in heterozygous female Virginia opossums (Didelphis virginiana) across a major portion of the developmental period. Panels of tissues obtained from fetuses, newborns, and pouch young were examined via polyacrylamide gel electrophoresis of the G6PD protein.

Ultraviolet radiation-induced skin lesions in laboratory opossums (Monodelphis domestica) exposed from the weanling stage.

After repeated exposure to ultraviolet radiation, adult laboratory opossums (Monodelphis domestica) may develop hyperplasia and neoplasia of the shaved skin. We exposed opossums from the weanling stage (8-10 weeks after birth) and compared the incidence of lesions at designated time points with those observed following introduction of adults (around 5 months old) to the same irradiation protocol (125 J/m2 of UVB three times each week for up to 40 weeks). The overall incidence of hyperplasia and neoplasia among opossums exposed from the weanling stage was markedly lower than among animals first exposed as adults, and hyperkeratosis and sarcoma were not observed.

Timetable of in vivo embryonic development in the grey short-tailed opossum (Monodelphis domestica).

The timing of development was examined in 496 embryos from female Monodelphis domestica collected at known time intervals after video recorded mating. Ovulation occurred approximately 20 hr (day 1) after mating, and fertilization was observed by 24 hr. Transport through the oviducts was rapid, and pronuclear stage embryos were recovered from the uterus as early as 24 hr after mating.

Malignant melanoma in ultraviolet irradiated laboratory opossums: initiation in suckling young, metastasis in adults, and xenograft behavior in nude mice.

Litters of suckling young of the laboratory opossum (Monodelphis domestica) were irradiated with UV light from sunlamps with a spectral emission peak at 302 nm (UVB) to induce melanocytic nevi. Total doses of 0.87-5.

[Prospects for obtaining a mapping panel for somatic cell marsupial-rodent hybrids for the short-tailed opossum Monodelphis domestica].

A possibility of obtaining a panel of marsupial-rodent somatic cell hybrid clones has been explored, with a view to mapping the genome of the opossum (Monodelphis domestica). Fusion of opossum cells (splenocytes, bone marrow cells, and fibroblasts) with fibroblasts of Chinese hamster or vole (HGPRT- and TK- mutants, respectively) produced 146 hybrid clones. The majority of marsupial-mammalian somatic cell hybrids were characterized by pronounced fragmentation and segregation of marsupial chromosomes.

Safety and effect of topical recombinant human interleukin-1beta in the management of pressure sores.

Interleukin-1beta is produced by numerous cell types including monocytes and fibroblasts. It has been shown to stimulate multiple cell types including fibroblasts, keratinocytes, endothelial cells, neutrophils, macrophages, and lymphocytes. Previously, interleukin-1beta was shown to accelerate healing in partial-thickness and full-thickness wounds in animals and was also shown to be safe when applied topically in Phase I human trials.

Microtubule configurations in oocytes, zygotes, and early embryos of a marsupial, Monodelphis domestica.

The marsupials represent a separate evolutionary lineage from eutherians from which they diverged over 100 million years ago. In order to explore the origin and mode of centrosome inheritance amongst this group of mammals, this study investigates the microtubule organization during fertilization, parthenogenesis, and polyspermy in the didelphid, Monodelphis domestica. Microtubules and DNA were visualized in maturing ovarian oocytes, parthenogenetically activated oocytes, monospermic and polyspermic zygotes, and early embryos.

Effects of NIDDM on lipoprotein(a) concentration and apolipoprotein(a) size.

We investigated the effects of non-insulin-dependent diabetes mellitus (NIDDM) on lipoprotein(a) (Lp[a]) and apolipoprotein(a) (apo[a]) in a population of Mexican-Americans. In plasma samples from 536 subjects, we measured Lp(a) concentrations, and we estimated apo(a) isoform sizes following immunostaining of plasma proteins resolved using sodium dodecyl sulfate electrophoresis. We identified 81 diabetic subjects who had 108 distinct apo(a) isoform bands.

Blood collection and surgical procedures for the laboratory opossum (Monodelphis domestica).

Procedures have been developed for collecting blood samples from the laboratory opossum (Monodelphis domestica) and for conducting survival surgery on adults. Blood samples up to 1 ml were collected routinely by cardiac puncture of animals anesthetized by inhalation of methoxyflurane or halothane with minimal risk; however, halothane required more careful monitoring. Surgical procedures such as partial hepatectomy unilateral hysterectomy, and tail wound repair were safely accomplished on animals anesthetized with methoxyflurane.

Susceptibility to ultraviolet-induced corneal sarcomas is highly heritable in a laboratory opossum model.

The laboratory opossum, Monodelphis domestica, develops hyperplasia and neoplasia of the corneal stroma after repeated exposure to low doses of ultraviolet radiation (UVR) (non-erythemic in skin). We exposed adult animals from genetically diverse families within our colony to determine whether there are any heritable components to the risk of this form of eye cancer. From about 5 months of age, animals were exposed 3 times a week to a dose of about 125 J/m2 of UVB (spectral peak = 302 nm).

Genetic analysis of ultraviolet radiation-induced skin hyperplasia and neoplasia in a laboratory marsupial model (Monodelphis domestica).

Monodelphis domestica, the laboratory opossum, develops hyperplasia and neoplasia of shaved skin after repeated exposure to ultraviolet radiation (UVR). We exposed Monodelphis from genetically diverse families within our colony to determine whether there are any heritable components to the risk of two distinct skin lesion phenotypes-melanocytic nevus (MN) and advanced hyperkeratosis (HK). From about 5 months of age, animals were shaved and exposed three times a week to a dose of about 125 J/m2 of UVR (spectral peak, 302 nm; range, 280-400 nm).

X-chromosome replication patterns in adult, newborn and prenatal opossums.

Somatic cells from the opossums Monodelphis domestica and Didelphis virginiana were labelled with 5-bromodeoxyuridine (BrdU), treated with colchicine, stained with acridine orange and examined using fluorescence microscopy. BrdU-incorporated metaphase spreads from females of M. domestica at developmental stages from late bilaminar blastocysts to adults showed replication asynchrony of the two (acrocentric) X chromosomes.

HPRT activity in embryos of a South American opossum Monodelphis domestica.

Marsupial females show preferential paternal X-inactivation. However, the time at which X-inactivation occurs in early development has not yet been determined. A double microassay which measures the activities of X-linked hypoxanthine phosphoribosyl transferase (HPRT) and the autosomally-coded adenine phosphoribosyl transferase (APRT) from the same sample was performed on a collection of embryos from a South American opossum Monodelphis domestica.

Cell lines derived from ultraviolet radiation-induced benign melanocytic nevi in Monodelphis domestica exhibit cytogenetic aneuploidy.

The gray short-tailed opossum, Monodelphis domestica, develops dermal melanocytic nevi (MN) after long-term chronic exposure to UVB (midwavelength ultraviolet radiation) alone. We developed cell lines from six UVB-induced dermal benign melanocytic lesion biopsies. One of the MN was determined histologically to be a benign melanoma (BM), whereas the remainder were benign melanocytic hyperplasias (MH).

A primate model of monotypism in atherosclerotic lesions.

We have characterized the expression of allelic variants of X-linked glucose 6-phosphate dehydrogenase (G6PD) in aorta from homozygous, hemizygous, and heterozygous baboons (Papio hamadryas). Fibrous plaques from heterozygous baboons fed a high cholesterol, saturated fat diet contained distributions of G6PD allelic variants that differed from those of normal arterial wall and fatty streaks. The skewed allelic expression patterns in fibrous plaques of heterozygotes reflect decreased cellular heterogeneity in advanced vascular lesions.

Baboon apolipoprotein A-IV. Identification of Lys76-->Glu that distinguishes two common isoforms and detection of length polymorphisms at the carboxyl terminus.

Various protein isoforms have been identified for human apolipoprotein A-IV (apoA-IV). However, investigations of their physiological effects have been limited because of low frequencies for many of the apoA-IV variants. Recent discovery of extensive variation in baboon apoA-IV using isoelectric focusing (IEF) makes this primate species an excellent model for genetic studies of apoA-IV.

Linkage between complement components 6 and 7 and glutamic pyruvate transaminase in the marsupial Monodelphis domestica.

The sixth and seventh components of complement were found to be polymorphic and tightly linked in the laboratory opossum (Monodelphis domestica), as they are in eutherian mammals. In addition, strong evidence for linkage of the C6-C7 haplotype to the gene for glutamic pyruvate transaminase (GPT) was obtained for females but not for males. This result, combined with previous observations, established as a generality that recombination is severely reduced in females of this species by comparison with males.

Blood groups in a marsupial (Monodelphis domestica).

We produced alloimmune sera against erythrocytes of the marsupial, Monodelphis domestica. By absorption fractionation, six operationally monospecific blood typing reagents (hemolysins) were prepared. Using segregation analyses, we found that each reagent detects a blood group antigen controlled by an autosomal dominant gene.

Structure and evolution of opossum, guinea pig, and porcupine cytochrome b genes.

We have sequenced the mitochondrial cytochrome b gene from the guinea pig, the African porcupine, and a South American opossum. A phylogenetic analysis, which includes 22 eutherian and four other vertebrate cytochrome b sequences, indicates that the guinea pig and the porcupine constitute a natural clade (Hystricomorpha) that is not a sister group to the clade of mice and rats (Myomorpha). Therefore, the hypothesis that the Rodentia is paraphyletic receives additional support.

Correlated expression of atrial myosin heavy chain and regulatory light chain isoforms with pressure overload hypertrophy in the non-human primate.

Objective: The aim was to determine the extent to which myosin heavy chain and light chain isoform transitions in atrial myocardium are coordinately regulated under pathological conditions in tissue from normal baboons, hypertensive baboons with myocardial hypertrophy, and baboons in which hypertrophy had regressed.

Methods: Quantitative distributions of myosin heavy chain (MHC) and regulatory myosin light chain (MLC2) isoforms in atrial myocardium from 35 adult baboons were determined by electrophoresis under denaturing conditions and laser densitometry.

Results: A significant association was observed between the ratios of MHC and MLC2 isoforms in atrial myocardium (r = 0.