Assignment of the bovine p53 gene (TP53) to chromosome 19q15 by fluorescence in situ hybridization.
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Animals↗
Biomedical subjects
M E Jackson
Publications and source records attributed to M E Jackson.
CSF-1 treatment promotes angiogenesis in the metaphysis of osteopetrotic (toothless, tl) rats.
It has recently been shown that following treatment with colony-stimulating factor-1 (CSF-1) the osteopetrotic condition in toothless (tl) rats greatly improves and growth is accelerated. We have examined the effects of such treatment on the microvasculature of the distal femoral chondro-osseous junction, a site where bone growth in length is coordinated with angiogenesis. Vascular casts and ultrastructural analyses of this region showed that, compared to untreated normal rats, untreated mutants showed little bone growth or angiogenesis. When mutants were treated with CSF-1 angiogenesis was markedly accelerated. These data show a remarkable effect of this growth factor on angiogenesis in this osteopetrotic mutation. Whether this effect of CSF-1 on angiogenesis is direct or indirect is not known and indicates that its effects on the normal microvasculature deserve further study.
Animals↗
Osteoblasts from the toothless (osteopetrotic) mutation in the rat are unable to direct bone resorption by normal osteoclasts in response to 1,25-dihydroxyvitamin D.
Osteopetrosis describes a diversified group of metabolic bone disorders characterized by a generalized, skeletal sclerosis resulting from reduced osteoclast-mediated bone resorption. The toothless (tl) osteopetrotic mutation in the rat is characterized by few osteoclasts and the inability to be cured by transplants of hemopoietic stem cells. This implies that the defect(s) responsible for reduced osteoclast activity in tl rats is within the skeletal microenvironment (cells or matrices). Osteoblasts and their products are known to play a role in regulating bone resorption and abnormalities in the osteoblast population in tl rats have been reported. The purpose of this study was to determine whether osteoblasts isolated from tl mutant rats, when cultured with normal osteoclasts, could increase bone resorption (pit formation) in response to stimulation by 1,25 dihydroxyvitamin D (1,25(OH)2D). The addition of 1,25(OH)2D produced a highly significant response in normal osteoblast cocultures but no response in mutant cultures. A dose response study with 1,25(OH)2D (10(-6) to 10(-9)M) revealed that mutant osteoblasts are unable to increase osteoclast activity. These data indicate that the vitamin D receptor-signal transduction pathway in tl rats needs to be examined.
Animals↗
Both viral E2 protein and the cellular factor PEBP2 regulate transcription via E2 consensus sites within the bovine papillomavirus type 4 long control region.
The bovine papillomavirus type 4 (BPV4) long control region (LCR) contains three consensus binding sites, E2(1), E2(2), and E2(3) (ACCN6GGT), for the viral E2 transcription factor and a fourth degenerate site, dE2 (ATCN6GGT), which lies 3 bp upstream of E2(3). The E2(2) site was found to bind the cellular transcription factor PEBP2, and mutations at this site reduced basal promoter activity by as much as 60%, indicating an important role for PEBP2 in LCR function. Mutation of the E2(3) or dE2 site slightly decreased basal promoter activity, but the cellular proteins binding these sites have not yet been characterized. E2 protein was found to have considerable influence upon LCR promoter activity in primary bovine palate keratinocytes. Thus, when high levels of BPV1 E2 were present, almost complete repression of the BPV4 LCR was observed, whereas smaller amounts of BPV1 or BPV4 E2 led to transactivation. Mutational analysis indicated that E2(1) and dE2 mediated transactivation by E2, whereas E2(2) and E2(3) were responsible for repression by E2. In vitro complexes of binding sites E2(1) and E2(2) with E2 protein demonstrated much greater stability than complexes formed by the E2(3) and dE2 sites. These data suggest that the four E2 sites in the BPV4 LCR each perform different functions in the control of transcription and that competition between cellular transcription factors and viral E2 proteins is essential in regulating the level of viral gene expression during papilloma development.
Animals↗
Transcriptionally active nuclei isolated from intact bone reflect modified levels of gene expression in skeletal development and pathology.
Transcriptional regulation of gene expression in vivo in bone, associated with normal development or skeletal disorders, to date, has not been studied. We report the successful isolation of nuclei that are transcriptionally active from normal and osteopetrotic rat bone. Transcription rates of cell growth and bone-related genes (including histone H4, c-fos, c-jun, TGF beta 1, beta 2 macroglobulin, collagen, fibronectin, osteocalcin, osteopontin, and tartrate resistant acid phosphatase) change as a function of calvarial development from birth to 6 weeks and are selectively modified in osteopetrotic animals. Additionally, nuclei isolated from intact bone yield promoter binding factors. Bone nuclei, which transcribe faithfully and contain the normal complement of nuclear protein factors, offer a powerful approach for investigating in vivo gene regulation in skeletal development and pathology.
Animals↗
Aberrant gene expression in cultured mammalian bone cells demonstrates an osteoblast defect in osteopetrosis.
Osteopetrosis is a skeletal condition in which a generalized radioopacity of bone is caused by reduced resorption of bone by osteoclasts. However, it has recently been shown that during skeletal development in several osteopetrotic rat mutations specific aberrations occur in gene expression reflecting the activity of the bone forming cells, osteoblasts, and the development of tissue organization. To evaluate their pathogenetic significance, progressive osteoblast differentiation was studied in vitro. Primary cultures of normal osteoblasts undergo a sequential expression of cell growth and tissue-related genes associated with development of skeletal tissue. We report that osteoblast cultures can be established from one of these mutants, toothless; that these cells in vitro exhibit similar aberrations in gene expression during cell proliferation and extracellular matrix formation and mineralization observed in vivo; and that an accelerated maturation sequence by mutant osteoblasts mimics the characteristic skeletal sclerosis of this disease. These data are the first direct evidence for an intrinsic osteoblast defect in osteopetrosis and establish an in vitro model for the study of heritable skeletal disorders.
Animals↗
Abnormalities of phosphoprotein gene expression in three osteopetrotic rat mutations: elevated mRNA transcripts, protein synthesis, and accumulation in bone of mutant animals.
Osteoclast abnormalities that characterize osteopetrosis, a disorder of bone resorption, may derive from aberrant signals from the osteoblast or the bone matrix. In the present studies, both synthesis and the bone matrix content of the major bone phosphoprotein component, osteopontin, were found to be elevated in three osteopetrotic rat mutations (ia, op, and tl). In whole bone, a twofold increase in the content of the characteristic amino acid O-phosphoserine for osteopontin occurred in op and tl mutant long bone, but a smaller (15%) and more variable increase was observed in ia mutant rat long bone. Extraction of the bone matrix components and partial purification by reverse phase chromatography showed a twofold increase in a phosphoprotein fraction relative to other noncollagenous components. Amino acid analysis and staining characteristics of SDS-PAGE fractionated proteins indicated this to be osteopontin. Organ cultures of calvarial bone from 4 day ia osteopetrotic mutant and normal rats in the presence of 3H-proline showed increased synthesis of this 60 kD protein, which was stimulated by vitamin D. Preparation of total cellular RNA from bone of 2- and 6-week-old mutants and normal rats supported increased synthesis of osteopontin as reflected by hybridization with osteopontin cDNA probe, showing significantly higher levels of mRNA transcripts in ia (3-5 fold), tl (1.4-2 fold), and op (6-25 fold) mutant bone compared to normal littermates. The changes in osteopontin mRNA levels in mutant bone were also examined in relation to other growth and phenotype-expressed genes. The findings of increased accumulation of osteopontin in osteopetrotic bone and increased synthesis by osteoblasts are interesting in light of the previously reported decrease in bone osteocalcin content (Endocrinology, 126:966, 1990), confirmed here by decreased osteocalcin mRNA transcripts. Such aberrations in the composition of skeletal extracellular matrix could be a reflection of or a contributing factor to the osteoclast abnormalities of some of these osteopetrotic disorders.
Animals↗
Measurements of the growth and distribution of mammalian cells in a hollow-fiber bioreactor using nuclear magnetic resonance imaging.
We have used diffusion-weighted 1H NMR micro-imaging and localized spectroscopy techniques to monitor the growth and distribution of mammalian cells in a hollow-fiber bioreactor. Non-invasive NMR measurements of this type should also allow investigation of metabolic heterogeneity and assist in future designs of hollow-fiber systems.
Cell Division↗
An element binding a C/EBP-related transcription factor contributes to negative regulation of the bovine papillomavirus type 4 long control region.
Deletion of the NR2 element of the long control region (LCR) of bovine papillomavirus type 4 (BPV-4) was observed previously to lead to a fivefold increase in enhancer activity of a subfragment of the LCR. Further characterization of this element indicates that mutations in NR2 lead to increased enhancer activity in both mouse CT3 fibroblasts and in a transformed bovine epithelial cell line derived from an alimentary canal papilloma/in situ carcinoma, but not in primary bovine keratinocytes. Since similar oligonucleotide-nuclear factor complexes were obtained in electrophoretic mobility shift assays (EMSA) for all three cell types, the observed difference in negative activity may result from variation in the NR2-binding factor itself between primary and established/transformed cell lines, or from the involvement of other factors that vary between the lines. Characterization of the NR2-binding factor by heat stability and antibody supershifts in EMSA indicate that the factor is related to the CCAAT/enhancer-binding protein (C/EBP) family, and that one component of the complexes may be C/EBP beta.
3T3 Cells↗
Correlates of disruptive behaviors in nursing homes. A reanalysis.
This study reexamines factors associated with the occurrence of disruptive behaviors using a representative sample of nursing home residents in Rhode Island in 1984-1985. Four indicators of disruptive behaviors are examined using multivariate methods: evidence of any disruptive behaviors, abusive behavior, wandering, and noisiness. Results are compared with the 1989 study by Jackson et al., which was based on the same data. Findings indicate that the likelihood of exhibiting disruptive behaviors in nursing homes increases with the severity of cognitive impairment, ADL dysfunction, and incontinence, and decreases with immobility. Immobility not only is negatively associated with wandering behavior, but with other behaviors as well. Women are less likely to be abusive. In contrast to the earlier study, which did not use multivariate methods, age and communication problems are not related to disruptive behaviors.
Activities of Daily Living↗
Evaluating the predictive validity of nursing home pre-admission screens.
This article demonstrates a method for evaluating the predictive validity of nursing home pre-admission screens (PAS) by using measures of predictive validity adapted from the field of epidemiology. Our approach estimates how well as PAS performs in identifying the "who but for" population of the Medicaid home and community-based services waiver programs for the frail elderly. The methodology's usefulness in screen revision is also illustrated.
Activities of Daily Living↗
Experimental studies of osteopetrosis in laboratory animals.
Osteopetrosis is a metabolic bone disease characterized by a systemic increase in skeletal mass. It results from a defect in the production or function of osteoclasts and is inherited in nine genetically distinct osteopetrotic animal mutations and man. Studies of these mutations have revealed that osteopetrosis is a complex, heterogeneous disorder in its expression, etiology, and response to treatment by bone marrow transplantation or by hormone/growth factor therapy. These animal mutations have been valuable tools for probing the pathogenesis and treatment of osteopetrosis, and information obtained from these studies has been used clinically for the treatment of humans with osteopetrosis. In addition, studies of these mutations have contributed significantly to understanding normal bone cell biology, including the origin of the osteoclast and the significance of colony-stimulating factor-1 in osteoclast development. The resistance of some of these mutations to cure by stem cell transplantation and hormone therapy, coupled with similar observations and experiences in the human condition, indicates that these animal mutations will continue to serve important roles in the development of alternative therapies to treat resistant forms of the disease. These studies are bound to improve the understanding of normal bone biology by providing additional insights into the regulation of osteoclasts by osteoblasts and their products or by other elements of the skeletal microenvironment.
Animals↗
Cooperation between papillomavirus and chemical cofactors in oncogenesis.
Papillomaviruses have been identified as causative agents of squamous cell carcinoma in humans and other mammals, and several papillomavirus-encoded proteins have been found to possess transforming activity. However, papillomavirus infection per se appears insufficient to elicit carcinomas in the majority of cases, and other agents have been implicated as cofactors. This review presents epidemiological studies linking environmental carcinogens to papillomavirus-associated carcinoma, and evidence from experimental systems that attempt to identify the targets of carcinogens and viral oncoproteins, and the interactions of both in progression to carcinoma.
Animals↗
Efficacy of nursing home preadmission screening.
This study evaluates the efficacy of four state nursing home preadmission screening instruments. Using data from three community-based services demonstration experiments, the screens were compared on measures of sensitivity, specificity, and proportions of false negatives and false positives. Results indicate that the more restrictive screens tended to have lower sensitivity but higher specificity, and to generate a higher proportion of false negative decisions than the more liberal screens. Overall rates of correct prediction were higher for the more restrictive screens. Results highlight differences in eligibility restrictiveness across states as well as possible tradeoffs of cost containment and access in designing a preadmission screen.
Aged↗
Myoelectric activity and sequencing of selected trunk muscles during isokinetic lifting.
Trained weight lifters lift heavy loads without a concomitant degree of acute low-back injuries. To study the process by which large loads are lifted with minimal injury, integrated electromyographic signals were recorded from four large muscle groups: gluteus maximus, quadriceps, latissimus dorsi, and erector spinae in 4 weight lifters and 11 asymptomatic control subjects. These signals were recorded during a floor-to-knuckle-height isokinetic lift (dead lift) at 30.5 and 45.7 cm/sec. The signals were normalized for the height of the lift and the maximal isokinetic integrated electromyographic activity. The weight lifters achieved maximal force at 50% of maximal lift height, whereas the control subjects achieved it at 67%. Although not statistically significant, the weight lifters used the gluteus maximus more during the early stages of the lift, perhaps contributing to earlier development of force. This process would stabilize the pelvis and permit the erector spinae to extend the trunk more efficiently. The weight lifter then completed the lift with prolonged and increasing activity in the quadriceps. This technique may minimize the required force in the erector spinae and the forces on the low-back structures. Clinical implications include more effective strength training of lifting muscle groups other than spinal extensors and the teaching of lifting strategies employed by weight lifters in low-back rehabilitation and work-hardening programs.
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Eligibility for publicly financed home care.
OBJECTIVES: Proposals for publicly financed home care for the elderly now tend to include cognitive impairment criteria as well as activities of daily living (ADL) criteria. The numbers of elderly deemed eligible for services will depend on the definitions of ADL and cognitive impairment used. METHODS: Data from the 1984 National Long-Term Care Survey were used to generate a series of estimates of the community-dwelling elderly with ADL disabilities and cognitive impairment. RESULTS: When only ADL criteria are used, estimates of disability range from 472,000 to over 3 million (1.6% to 12.5% of the community-dwelling elderly). These estimates increase to approximately 1 million to 4.2 million (3.5% to 14.0% of the community-dwelling elderly) when cognitive impairment criteria are added. CONCLUSIONS: The use of more stringent or more liberal eligibility criteria will have dramatic effects on the number of elders who qualify for services. The nature of the eligibility criteria employed in any expansion of federally financed home care benefits will be a major factor in determining the costs of such a program.
Activities of Daily Living↗
Performance of growing pullets and laying hens fed low-protein, amino acid-supplemented diets.
An experiment was conducted to determine the effect of feeding low-protein, amino acid-supplemented diets during growing and laying periods on performance of a commercial strain of White Leghorn chickens. The birds of the positive control were fed diets in a sequence of 20, 16, and 14% protein during 0 to 6, 6 to 12, and 12 to 18 wk (growing period) and of 18, 16.5 and 15% protein during 18 to 34, 34 to 50, and 50 to 66 wk of age (laying period). The birds of the negative control were fed protein levels of 16, 13.5, and 11.5% in the growing period and 14, 13, and 12% in the laying period. Other groups were fed the negative control series supplemented with methionine, methionine plus lysine, or methionine plus lysine plus other deficient essential amino acids. An additional group was fed the negative control supplemented with methionine plus lysine during the growing period and a protein sequence of 15, 14, and 13% supplemented with methionine and lysine during the laying period. At 18 wk of age, birds fed the negative control supplemented with methionine plus lysine or methionine plus lysine plus other deficient essential amino acids had comparable body weight to those fed the positive control despite significantly lower protein and lysine intake. Overall egg production and egg weight of birds fed the sequence of 14, 13, and 12% protein supplemented with combination of methionine, lysine, and extra levels of tryptophan and isoleucine or of the birds fed the sequence of 15, 14, and 13% protein supplemented with methionine and lysine were not different from those fed the positive control. However, egg mass and body weight were inferior to those of birds fed the positive control.
Amino Acids↗
Gene expression during skeletal development in three osteopetrotic rat mutations. Evidence for osteoblast abnormalities.
Osteopetrosis is a group of metabolic bone diseases characterized by reductions in osteoclast development and/or function. These aspects of osteoclast biology are known to be influenced by osteoblasts and their products. To ascertain whether osteoblast dysfunction contributes to aberrations in the structural and functional properties of osteoclasts in osteopetrosis, we systematically examined gene expression as reflected by mRNA levels for a series of cell growth- and tissue-related genes associated with the osteoblast phenotype during skeletal development in normal and mutant rats of three different osteopetrotic stocks. We show that the methods used permit the reproducible isolation of undegraded total cellular RNA from bone and that mRNA levels can be reliably quantitated in these preparations. Each osteopetrotic mutation exhibits a distinct aberrant pattern of osteoblast gene expression that may be correlated with and explain some abnormalities in extracellular matrix composition, mineralization, osteoclast development, and effects of elevated serum levels of 1 alpha,25-dihydroxyvitamin D3, depending upon the mutation. Normal rats show minor variations in gene expression that reflect the genetic background (stock). This, the first comprehensive molecular analysis of osteoblast gene expression in osteopetrosis, suggests that some osteopetroses, particularly in the toothless rat, are associated with and potentially related to mechanisms associated with aberrations in osteoblast function. More generally, the present studies demonstrate alterations in gene expression as reflected by mRNA levels that are associated with functional properties of the osteoblast, particularly those contributing to the recruitment and/or differentiation of osteoclasts, thereby influencing skeletal modeling.
Animals↗