Biomedical subjects

J Stafford

Publications and source records attributed to J Stafford.

41 records · Page 3Linked to original sources

PubMedDate not supplied

Inhibition of macrophage activity by mitogen-induced goldfish leukocyte deactivating factor.

Macrophage activating and deactivating cytokines have been characterized in mammalian systems but little is known about these immunoregulatory molecules in fish. Using gel permeation and chromatofocusing fast performance liquid chromatography (GP-FPLC and C-FPLC) we partially purified a macrophage deactivating factor (MDF) from mitogen-induced goldfish kidney leukocytes. Inhibition of the macrophage-derived nitric oxide (NO) response induced by this MDF was time-, dose- and temperature-dependent. Macrophages pre-treated for 6 or 24 h with MDF before activation with macrophage activating factors (MAF) and/or bacterial lipopolysaccharide (LPS) exhibited a down-regulation in their NO response, while those treated with MDF 24 h after activation with MAF and LPS did not. MDF treatment also impaired the NO response of goldfish macrophages infected with the mammalian protozoan parasite Leishmania major. These results suggest that MDF exhibits its inhibitory effect downstream of the converging intracellular pathways induced by LPS and/or L. major. The novel teleost MDF has an approximate Mr of 15 kD and a pI < 4, and is the first endogenous molecule of teleosts known to down regulate macrophage antimicrobial responses.

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PubMedDate not supplied

Cell-type specific expression of a transfected immunoglobulin gene.

The introduction of cloned genes into eukaryotic cells has become a major technique in the study of gene expression. Many experiments have demonstrated transcription of cloned genes after transfection into heterologous cell systems--cells in which the genes are not normally active. More recently, several investigators have obtained expression of specialized genes after transfection into cells of the corresponding specialized type, notably the beta-globin gene in erythroleukaemic cells and immunoglobulin genes in myeloma cells. These results allow the study of gene expression during development by comparing transcription of a gene transfected into homologous and heterologous cells. We have shown that a rearranged kappa immunoglobulin gene, cloned from a mouse myeloma, is transcribed transiently at a high level when reintroduced into mouse myeloma cells. We show here, in an internally controlled manner, that the same immunoglobulin gene is not detectably transcribed when transfected into mouse 3T3 or L cells.

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PubMedDate not supplied

An immunoglobulin promoter displays cell-type specificity independently of the enhancer.

Recent studies in which cloned immunoglobulin genes were introduced into cultured cells have produced two significant findings. First, the genes are expressed after transfection into lymphoid cells but not non-lymphoid cells. Second, transcription of an immunoglobulin gene requires, in addition to the promoter region, an enhancer element located downstream of the transcription start site. These findings raise the question of whether it is the promoter or the enhancer region that is responsible for the observed cell-type specificity. It has, in fact, been shown that immunoglobulin enhancers function only in lymphoid cells. We show here that the promoter for an immunoglobulin kappa light-chain gene also is strongly specific for lymphoid cells. Our result reemphasizes the importance of promoters relative to enhancers in determining which cells express which genes.

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PubMedDate not supplied

Synergism between immunoglobulin enhancers and promoters.

Enhancers are DNA sequences that stimulate transcription from eukaryotic promoters. This stimulatory effect can be exerted over large distances and from a position either 5' or 3' of a promoter. Enhancers have been found in the genomes of many viruses, and in some cellular genes such as those encoding immunoglobulin heavy chain and kappa light chain. An important feature of both viral and cellular enhancers is the ability of each enhancer to stimulate transcription from many promoters other than the one with which it is found associated. However, the question of whether cellular enhancers stimulate their 'own' promoter more efficiently than other promoters has apparently not been investigated. We show here that the kappa light-chain enhancer stimulates a kappa promoter about 20-fold more than it stimulates either the simian virus 40 (SV40) early promoter or a metallothionein (MT) promoter, two promoters that are very sensitive to other enhancers. Similarly, the heavy-chain enhancer stimulates a heavy-chain promoter much more than it stimulates the SV40 and MT promoters. This synergism between immunoglobulin enhancers and promoters might be due to the action of a protein that binds specifically to each of the regulatory elements.

Animals↗

PubMedDate not supplied

Use of computerized CT analysis to discriminate between Alzheimer patients and normal control subjects.

A newly developed computerized technique was used to analyze the CT scans of 49 patients with dementia of the Alzheimer type and 31 normal control subjects. Nine brain regions distributed across five CT slices were evaluated for each individual. For the purpose of analysis, the patients and controls were divided into an exploratory set and a test set. Several discriminant functions were conducted on the exploratory set and applied to the test set. The combination of variables that focused on regions in the temporal lobe was most accurate in differentiating Alzheimer patients from controls (94%). This degree of accuracy was achieved only when subjects younger than 65 years old were analyzed separately from those 65 years old and older. The newly developed computer software program was able to discriminate between independently selected groups of Alzheimer patients and control subjects. The program was most effective when the analysis emphasized regions in the temporal lobe and when subjects younger than 65 years old were analyzed separately from those 65 years old and older.

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