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L Carlson

Publications and source records attributed to L Carlson.

At least 55 records · Page 3Linked to original sources

Ongoing diversification of the rearranged immunoglobulin light-chain gene in a bursal lymphoma cell line.

The chicken immunoglobulin light-chain gene (IgL) encodes only a single variable gene segment capable of recombination. To generate an immune repertoire, chickens diversify this unique rearranged VL gene segment during B-cell development in the bursa of Fabricius. Sequence analysis of IgL cDNAs suggests that both gene conversion events derived from VL segment pseudogene templates (psi VL) and non-template-derived single-base-pair substitutions contribute to this diversity. To facilitate the study of postrecombinational mechanisms of immunoglobulin gene diversification, avian B-cell lines were examined for the ability to diversify their rearranged IgL gene during in vitro passage. One line that retains this ability, the avian leukosis virus-induced bursal lymphoma cell line DT40, has been identified. After passage for 1 year in culture, 39 of 51 randomly sequenced rearranged V-J segments from a DT40 population defined novel subclones of the parental tumor. All cloned V-J segments displayed the same V-J joint, confirming that the observed diversity arose after V-J rearrangement. Most sequence variations that we observed (203 of 220 base pairs) appeared to result from psi VL-derived gene conversion events; 16 of the 17 novel single nucleotide substitutions were transitions. Based on these data, it appears that immunoglobulin diversification during in vitro passage of DT40 cells is representative of the diversification that occurs during normal B-cell development in the bursa of Fabricius.

Alleles↗

Copper and very low birthweight babies.

Serum concentrations of copper and ceruloplasmin were measured in 24 very low birthweight babies. They were high in those weighing less than 1000 g, and critically ill or receiving intravenous nutrition, and normal in those with bone disease, neutropenia, or oedema. Care is needed to maintain adequate concentrations without toxicity.

Ceruloplasmin↗

Quantitative inter-strain comparison of the distribution of choline acetyltransferase activity in the rat cochlear nucleus.

The distribution of choline acetyltransferase activity in the cochlear nucleus of Sprague-Dawley albino rats was quantitatively compared to those in two strains of pigmented rats, Long Evans hooded and Brown Norway, using microdissection and radiometric assay techniques. Although activities tended to be, on the whole, higher in the albino rats, the differences were fairly minor. The relative distributions of choline acetyltransferase activity were generally similar among the 3 rat strains, not only among regions, but also within regions. Stain for acetylcholinesterase activity in the cochlear nucleus also had a similar appearance among the 3 rat strains. These chemical results are consistent with previous anatomical and physiological studies suggesting that auditory differences between albino and pigmented animals may not be as great in the cochlear nucleus as in the superior olivary complex.

Animals↗

Quantitative analysis of the T cell response to antigen and planar membranes containing purified Ia molecules.

Planar lipid membranes containing the purified Ia molecule E beta k:E alpha k can present a peptide antigen derived from cytochrome c to the T cell hybridoma 2B4.11. The incorporation of E beta k:E alpha k into planar membranes was linear over a 120-fold range of Ia molecule concentrations, permitting the dependence of the T cell response on the Ia molecule concentration to be examined. As the Ia molecule concentration was increased in the planar membranes, two parameters changed: less antigen was needed to stimulate the T cells, and the plateau response seen at functionally saturating concentrations of antigen increased. The antigen sensitivity was analyzed by plotting the antigen concentration (log2) required to stimulate the release of 10 U of IL 2 from the T cells as a function of the Ia molecule concentration (log2). If the T cell recognized a simple unit of one antigen molecule and one Ia molecule, this plot should have generated a straight line with a slope of -1. Surprisingly, a line with a slope of -2.04 X/divided by 1.12 was observed, suggesting that the T cell might recognize one antigen molecule and two Ia molecules. This complexity, however, resulted from changes in the maximal response achieved at different Ia molecule concentrations. A similar phenomenon was observed when the Ia molecule concentration was decreased in cultures containing splenic antigen-presenting cells (APC) by the addition of an anti-E beta k:E alpha k monoclonal antibody, or the use of [B10.A(4R) X B10.PL]F1APC. The Ia molecule concentration can therefore be limiting for T cell hybridomas in cultures containing normal APC and functionally saturating amounts of antigen. When the planar membrane data were normalized to the maximal response to eliminate the effect of the changing plateau response, the resulting plot generated a line with a slope of -1.17 X/divided by 1.11. These results suggest that the sole stimulatory signal for this T cell hybridoma consisted of a 1:1 ratio of antigen and Ia molecules.

Algorithms↗

Optimization of antigen presentation to T cell hybridomas by purified Ia molecules in planar membranes. Ia molecule polymorphism determines the antigenic fine specificity of the response to cytochrome c peptides.

Ia molecule (Ek,b beta:Ek alpha or Ak beta:Ak alpha)-containing planar membranes were constructed with cholesterol and a 9:1 molar ratio of the phospholipids dipalmitoyl phosphatidylcholine and dilinoleoyl phosphatidylcholine. This lipid composition was found to be optimal for the stimulation of T cell hybridomas of different specificities. Use of this system allowed the detection of weak responses not measurable when other artificial membranes were used. Activation of the cytochrome c, Ek,b beta:Ek alpha-reactive hybridoma 2B4.11 using such membranes resulted in responses comparable to those found using antigen-presenting cells (APC); that is, similar amounts of IL-2 were produced at the same concentrations of antigenic peptides. Presentation of moth and pigeon cytochrome c peptides by Ek beta:Ek alpha- or Eb beta:Ek alpha-reconstituted membranes resulted in 2B4.11 response patterns similar to those previously described using B10.A or B10.A(5R) APC. These data conclusively demonstrate that differential stimulation by moth and pigeon cytochrome c peptides depends solely on structural differences in the E beta:E alpha molecules used for antigen presentation.

1,2-Dipalmitoylphosphatidylcholine↗

Treatment of reflex sympathetic dystrophy of the hand with an active "stress loading" program.

Reflex sympathetic dystrophy (RSD) is a syndrome characterized by pain out of proportion to injury, vasomotor and trophic changes, stiffness, and decreased function. It is important to separate the active disease process of RSD from its resultant state of contracture and fibrosis. Our treatment program is based on active "stress loading," which consists of active traction and compression exercises that provide stressful stimuli to the extremity without joint motion. Fifty-two patients with RSD were treated during a 3-year period. Their results and long-term follow-up on 41 patients are presented. The "stress loading" program has been used consistently during the past 20 years. The advantages of the program are its effectiveness, simplicity, safety, and noninvasiveness.

Adult↗

Confirmation of Legionella pneumophila cultures with a fluorescein-labeled monoclonal antibody.

We compared a fluorescein-labeled monoclonal antibody directed against an outer membrane protein of Legionella pneumophila (Genetic Systems Corp. [GSC], Seattle, Wash.) with a similarly labeled polyclonal reagent (L. pneumophila serogroups 1 to 6, poly; BioDx, Inc., Denville, N.J.) for the confirmation of L. pneumophila isolates grown in culture. Duplicate suspensions of 52 organisms, including 21 L. pneumophila and 8 non-L. pneumophila species of legionella, were placed on individual glass slides, fixed, and stained with both reagents, and the results were compared. Both antisera correctly identified all L. pneumophila serogroups 1 to 6, but only the GSC reagent produced definitive staining of the L. pneumophila isolates of serogroups 7, 8, and 9. Additionally, the GSC reagent produced more uniform staining patterns around the legionella bacilli and displayed little background fluorescence when compared with the BioDx reagent.

Antibodies, Bacterial↗

Instruments for rapid cytophotometric analysis of morphologically identified clinical cell material.

Two instruments for cytophotometric analysis are described. The first is a generally applicable object stage scanning microspectrophotometer for the wavelength region 240-700 nm. The microscopic image is shown on a monitor screen and delineation of the analyzed objects is made with the aid of a light pen. The second instrument is a high speed microscope photometer specially adapted for the measurement of DNA, nuclear protein, and projected nuclear area following combined Feulgen and Naphthol Yellow S staining. A two-dimensional CCD sensor measures the light intensity in the microscopic image with a resolution of 0.3 micron. The objects are selected by the operator pointing the light pen at the image screen. The integrated absorbance at two different wavelengths, together with the projected area, is automatically measured for each object and presented in histogram form. Cells in clusters or sections can be measured under operator control with the help of the light pen. The time required for measuring 100 cells is 5-10 min in an ordinary preparation. The application of these instruments to the grading of cellular atypia and in the prognostic evaluation of malignant tumors is demonstrated on material from developing squamous bronchial carcinoma and in invasive mammary and prostatic adenocarcinoma.

Cells↗

Pigment mutations associated with altered circadian rhythms in mice.

Mammalian albinism is known to alter neural pathways, reduce retinal pigment, and affect diverse behaviors. Mammalian circadian rhythms have been shown to depend on visual pathways, respond to light intensity and regulate many behaviors. Here we show that mice homozygous for the recessive albino or pinkeye-dilute mutations display shorter circadian rhythms than pigmented controls. We conclude that these pigment loci, or closely linked loci, influence the expression of circadian rhythms in mice.

Animals↗