Search PubMed⌕ Search

Biomedical subjects

J F Watkins

Publications and source records attributed to J F Watkins.

At least 37 records · Page 2Linked to original sources
PubMed1989

Gender and race differentials in elderly migration.

A number of studies have provided evidence that elderly migration in the United States is strongly selective in terms of the characteristics of the migrants. Such characteristics as age, gender, and race are commonly used in examinations of migrant selectivity, but the differences in spatial behavior of these subpopulations are still poorly understood. This article uses state-to-state migration data, categorized by age, gender, and race, to explore the comparative patterns of origins, destinations, and migration propensities, with a focus on the southeastern states of the country.

Aged↗
PubMed1988

Excision repair of UV damage in human fibroblasts reversibly permeabilized by lysolecithin.

We have examined nucleotide excision repair synthesis in confluent human diploid fibroblasts permeabilized with lysolecithin. Following a UV dose of 12 J/m2, maximal incorporation of [alpha 35S]dNTPs occurred at a lysolecithin concentration (approximately 80 micrograms/ml) where slightly more than 90% of the cells were initially permeable to trypan blue. However, autoradiography of cells, permeabilized at this lysolecithin concentration, demonstrated that only about 20% of the total cell population incorporated significant levels of 35S into DNA. This result presumably reflected the fact that approximately 20% of the total cell population remained permeable for much longer periods of time (up to 2 h) than the remaining cell population (less than 20 min). The incorporation of dNTPs by UV-irradiated, permeabilized cells appeared to be bona fide excision repair synthesis since: (1) Incorporation was completely absent in unirradiated, permeabilized cells and in irradiated, permeabilized repair-deficient cells. (2) Nucleotides incorporated in the presence of BrdUTP were associated with normal density DNA. (3) The apparent Km for all 4 dNTPs was 50-100 nM, in agreement with past reports on human fibroblasts irreversibly permeabilized by cell lysis. (4) DNA associated with the newly incorporated dNTPs underwent ligation and rearrangements in chromatin structure analogous to what is observed in intact human cells. Repair incorporation of dNTPs was rapid and linear during the first 2 h after UV irradiation and permeabilization. After this time, incorporation ceased or continued at a much slower rate. Cell viability experiments and autoradiography demonstrated that the cells permeabilized to [3H]dNTPs were capable of carrying out DNA replication and cell division. Thus, confluent human diploid fibroblasts can be reversibly permeabilized to labeled dNTPs by lysolecithin for the study of excision repair following physiologic doses of UV radiation. However, under these conditions, only a fraction of the cells remain permeable for an extended period of time.

Cell Membrane Permeability↗
PubMed1985

Nucleosome rearrangement in vitro. 1. Two phases of salt-induced nucleosome migration in nuclei.

We have investigated the salt- and temperature-induced rearrangement of nucleosomes in both intact and H1-depleted nuclei from human cells. In agreement with previous reports on the rearrangement of nucleosomes in isolated chromatin or chromatin fragments, we observed a decrease in the average nucleosome repeat length following incubation of nuclei at 37 degrees C in elevated salt concentrations. However, this decrease occurred in two distinct phases. First, incubation of H1-depleted nuclei at 37 degrees C for as little as 10 min in low-salt, isotonic buffer (containing 0.025 M KCl) resulted in a shift in the limiting repeat value from approximately 190 to 168 base pairs (bp). A similar shift was observed for intact nuclei incubated at 37 degrees C for 1 h in buffer containing near-physiological salt concentrations (i.e., 0.175 M KCl). This limiting repeat value was maintained in both intact and H1-depleted nuclei up to a salt concentration of 0.45 M KCl in the incubation buffer. Second, at salt concentrations of 0.625 M KCl, a limiting repeat of approximately 146 bp was obtained, and the nuclei had clearly lysed. During the first shift in repeat length, little additional exchange of nuclear proteins occurred compared to nuclei kept on ice in a low-salt buffer. This was the case even though the conditions used to monitor exchange were optimized by using a high DNA to chromatin ratio. On the other hand, a significant increase in the exchange of nuclear proteins, and formation of nucleosomes on the naked DNA, was observed during the shift in repeat length to 146 bp.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Radioisotopes↗
PubMed1985

Nucleosome rearrangement in vitro. 2. Formation of nucleosomes in newly repaired regions of DNA.

We have reported previously that immediately following nucleotide excision repair in human cells the newly repaired DNA lacks a nucleosome conformation [Smerdon, M. J., & Lieberman, M. W. (1980) Biochemistry 19, 2992-3000]. In this study, we have examined the ability of these nascent DNA regions to acquire a nucleosome structure in vitro by incubating intact or H1-depleted nuclei in buffers containing different salt concentrations (0.025-0.625 M KCl) at 0 or 37 degrees C. Nucleosomes were detected in these regions by an increase in the level of repair-incorporated nucleotides associated with isolated nucleosome core particle DNA. Our results indicate that the nascent DNA is resistant to nucleosome formation during the low-salt transition where the limiting repeat length decreases from approximately 190 to 168 base pairs (bp) [Watkins, J. F., & Smerdon, M. J. (1985) Biochemistry (preceding paper in this issue)]. This result provides further evidence that the nascent DNA is indeed in a nonnucleosomal state. At higher salt concentrations (greater than 0.4 M), where the nucleosome repeat length decreases to a limiting value of approximately 146 bp, there was an increase in nucleosome formation in nascent DNA that correlated with the decrease in limiting repeat length. However, we did not observe a complete randomization of the repair-incorporated nucleotides. Indeed, even at the highest salt concentration used (0.625 M), we never observed more than 50% of the nascent DNA associated with the isolated core particles. This was the case even though a major portion of the nucleosomes had a limiting value repeat length following the high-salt incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Radioisotopes↗
PubMed1982

Studies on a human melanoma x hamster hybrid line selected by metastasis.

A highly metastatic hybrid line of human melanoma and tumorigenic, poorly metastatic hamster cells (BHK-21) was selected by pulmonary metastasis after injection of a fusion mixture into a Syrian hamster. The in vivo growth properties, karyotype, and plasminogen activator production of the line are reported. Monoclonal antibodies against the line indicated that it was expressing at least 8 human antigens although no melanoma-specific antigens were detected.

Animals↗
PubMed1980

Persistent infection of a cell line of mouse origin after cell fusion by u.v.-inactivated Sendai virus.

A cell line derived from Sendai virus-induced fusion of human adenocarcinoma and CBA mouse embryo cells had Sendai virus antigen (detected by immunofluorescence), together with bi-armed marker chromosomes, in 100% of the cells. After repeated passage, antigen-free cells carrying the same marker chromosomes appeared in the culture. Acrylamide gel analysis showed that all the Sendai virus antigens of antigen-positive cells were normal with the exception of the M protein. Antigen-negative cells contained no virus proteins and could be superinfected with wild-type virus, when all virus proteins appeared.

Adenocarcinoma↗
PubMed1978

Tumour-necrosis factor from the rabbit. I. Mode of action, specificity and physicochemical properties.

Sera from rabbits injected with BCG and then with endotoxin contain a factor (tumour-necrosis factor TNF) which, even at high dilutions, is cytotoxic in vitro for mouse L cells and some other cell lines. Using a 51Cr-release assay, cytotoxicity was detected as early as 7-8 h after addition of TNF serum to L cells and cell death was evident microscopically by 24 h. TNF was cytotoxic at 37 degrees C but not at 21 degrees C or 4 degrees C, and acted on both dividing and non-dividing cells. The antimetabolites sodium azide and dinitrophenol partially protected L cells from TNF, suggesting that actively metabolizing cells are the most sensitive. Treatment of L cells with trypsin did not delay cytotoxicity nor was cytotoxicity inhibited in the presence of various saccharide derivatives of cell-surface glycoproteins. Rabbit TNF was remarkably stable with a mol. wt. of 40-50,000. It was eluted with the more acidic serum proteins on ion-exchange chromatography, but precipitated in 50%-saturated ammonium sulphate. Sensitivity to TNF could not be correlated with tumourigenicity of several animal and human lines tested nor with the production of C-type viruses.

Animals↗
PubMed1977

Properties of a cell line from human adenocarcinoma of the rectum.

A new, highly differentiated line of cells derived from adenocarcinoma of the rectum (HT55) is described. This line is noteworthy for the following features: 1. The role played in its development by the use of UV-inactivated Sendai virus to attach tumour cell clumps to plastic bottles. 2. Evidence that it produces RNA-containing material of density 1-5--1-16 g/ml. 3. Induction of bone formation in the stroma when grown in athymic mice. 4. Stimulation of primary CBA mouse embryo fibroblasts to form a transient nodule when mixed with them and injected into adult CBA mice. The karyotype and growth-cycle characteristics of the line are described.

Adenocarcinoma↗