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Biomedical subjects

S L Perkins

Publications and source records attributed to S L Perkins.

At least 73 records · Page 4Linked to original sources

T-cell lymphocytosis associated with invasive thymomas.

Peripheral T-cell lymphocytosis and bone marrow infiltration is a rarely associated feature of invasive thymomas. Hypotheses for the origin of the circulating lymphocytes include a spillover of tumor lymphocytes into the circulation, a neoplastic lymphoid proliferation, and a reactive proliferation of peripheral lymphocytes resulting from a thymoma-induced immunoregulatory disorder. The authors describe two cases of peripheral T-cell lymphocytosis associated with invasive thymomas with a predominantly lymphocyte-rich, cortical type histologic appearance. In both cases T cells constituted as much as 99% of the circulating lymphoid cell population, according to flow-cytometric analysis before and after treatment, and expressed a mature phenotype consistent with medullary thymic or peripheral T cells (CD2+, CD3+, CD4+ or CD8+, CD5+, CD7+) with predominant expression of the alpha/beta T-cell receptor heterodimer. Bone marrow biopsy specimens showed nodular and interstitial infiltrates of small T cells. No monoclonal rearrangement of the T-cell receptor beta gene was observed. Both cases had a normal female karyotype. Our findings confirm those of previous studies supporting a reactive origin for the peripheral lymphocytosis. Although an immunoregulatory disorder is probably involved in lymphocyte proliferation in situ, we postulate that peripheral lymphocytosis results from augmented release of the more mature thymoma-associated lymphocytes into the circulation through tumor invasiveness. A thymic origin is supported by the presence of a subset of peripheral T cells with low-intensity CD5 expression.

Aged↗

Immunohistochemical staining for DNA topoisomerase II in frozen and formalin-fixed paraffin-embedded human tissues.

DNA topoisomerase II is the molecular target of several clinically useful chemotherapeutic drugs. The sensitivity of cells to drugs that target topoisomerase II is dependent on the cellular content of this enzyme. Drug-sensitive cells have elevated amounts of type II topoisomerase. To determine relative amounts of enzyme in malignant neoplasms, we developed an in situ immunohistochemical stain for topoisomerase II. The stain uses either polyclonal or monoclonal antibodies produced against the alpha isoform of the enzyme. Staining can be done on both frozen and formalin-fixed, paraffin-embedded tissues. By using this immunostain, we found marked differences in enzyme content in several human malignancies.

Animals↗

Anaplastic large-cell lymphoma presenting primarily in bone in a patient with AIDS.

Non-Hodgkin's lymphomas have been recognized as an important and frequently fatal part of the spectrum of diseases associated with HIV infection. These are most often high-grade B-cell lymphomas usually of immunoblastic and small cell non-cleaved subtypes. Sporadic reports of T-cell lymphomas associated with HIV infection are found in the literature. Two have been reported to be CD30 positive presenting with lymph node and skin involvement. We report a case of an AIDS patient with a T-cell anaplastic large-cell lymphoma that was CD30-positive and presented in bone. This is most probably a sporadic event rather than another part of the AIDS-associated spectrum of disease.

Bone Neoplasms↗

Interleukin-4 enhances murine bone marrow macrophage M-CSF receptor expression by a posttranscriptional mechanism.

Activated T-lymphocytes secrete interleukin-4 (IL-4), which has been shown to modulate a variety of monocyte activities requiring monocyte/macrophage colony-stimulating factor (M-CSF). To account for this interaction, we postulated that IL-4 acts on target cells by altering the expression of the M-CSF receptor (M-CSFr). To test this hypothesis, murine bone marrow macrophages were cultured under conditions that down-regulate M-CSFr and the effect of IL-4 on the reexpression of the receptor measured by binding of 125I-labeled M-CSF to the cells. The data show that incubation with IL-4 results in a dose-dependent, 2-3 x increase in M-CSFr with no change in binding affinity and a maximal effect on binding at about 12 h. This increase in M-CSFr is dependent upon new, specific protein synthesis as shown by the inhibitory action of cycloheximide, and gel analysis of radiolabeled, specific protein, immunoprecipitated with anti-M-CSFr antibody. Treatment with IL-4 does not stimulate M-CSFr mRNA expression but, consistent with enhanced receptor levels, does result in a heightened proliferative response to M-CSF. Thus, IL-4 affects M-CSF treated monocytic cells, at least in part, by altering the expression of M-CSFr.

Animals↗

Regulation of mouse bone marrow macrophage mannose receptor expression and activation by prostaglandin E and IFN-gamma.

The macrophage mannose receptor mediates the clearance of microorganisms and glycoproteins containing terminal mannose oligosaccharides. Cell surface expression of this receptor progresses with macrophage differentiation, and thus may be critical to the scavenger function of tissue and circulating macrophages. Bone marrow macrophages, which were used in this study, differentiate in culture and express functional mannose receptors. The cytokine IFN-gamma triggered activation of these macrophages and down-regulated cell surface expression of the mannose receptor after 48 h. Macrophage activation, as assessed by the generation of superoxide radicals, was inversely correlated with mannose receptor expression. The number of surface receptors was diminished by exposure to IFN-gamma, whereas the binding affinity of the mannose receptor remained unchanged. Treatment with IFN-gamma reduced receptor biosynthesis yet did not alter receptor degradation. Mannose receptor biosynthesis is up-regulated by PG of the E series, and these anti-inflammatory agents reversed the effects of IFN-gamma on receptor expression. Down-regulation of the mannose receptor by IFN-gamma was fully reversible by PGE, indicating that receptor levels are dependent on the functional state of the cell rather than being linked to terminal cell differentiation. The regulation of the receptor by cytokines and anti-inflammatory reagents suggests that the mannose receptor plays a critical role in macrophage scavenger functions and potentially in modulating inflammatory reactions.

Animals↗

A rapid high-performance thin-layer chromatographic urine screen for laxative abuse.

Laxative abuse, if unrecognized, can lead to unnecessary and costly investigations. The stimulants and cathartics are the most commonly abused laxatives and have the potential for causing the most long-term damage. We have optimized a solid-phase extraction coupled with high-performance thin-layer chromatography (HPTLC) to provide rapid, simple, and sensitive detection of phenolphthalein, danthron, rhein, aloin, and bisacodyl and its metabolites in urine. Positive screens have revealed laxative abuse in 3 of 42 samples from patients presenting with diarrhea of unknown origin.

Anthraquinones↗

Immune complex glomerulonephritis with unusual microfibrillar deposits associated with primary bone marrow lymphoma.

Glomerular microfibrillary deposits are characteristic of several diseases of the kidney. In a number of glomerulopathies, the nature of these microfibrillary deposits is critical in classifying the renal lesion and in suggesting the possibility of an associated systemic process. However, it is likely that as efforts are made to classify glomerulopathies with microfibrillary deposits, certain cases will defy categorization. We describe one such case in which a patient presented with rapidly progressive glomerulonephritis associated with large subepithelial, parallel-arrayed microfibrillar deposits associated with a primary bone marrow B-cell lymphoma. While IgG, C3, and lambda and kappa light chains were deposited in the glomerulus, serum and urine protein electrophoresis were normal. Treatment with Cytoxan and prednisone caused simultaneous remission of the lymphoma and the glomerulonephritis. Relapse of the lymphoma was associated with rapid deterioration of renal function. This case may represent a newly described variant of immune complex-mediated glomerulonephritis associated with microfibrillary deposits. The possibility is raised that the glomerular lesion is due to atypical immunoglobulins synthesized by a bone marrow lymphoma.

Biopsy↗

The failure of newborn mice infected with Escherichia coli to accelerate neutrophil production correlates with their failure to increase transcripts for granulocyte colony-stimulating factor and interleukin-6.

We quantified circulating and storage neutrophils, their precursors and progenitors, and mRNA for some of the cytokines involved in granulocytopoiesis, in newborn and adult mice following intrapulmonary inoculation of Escherichia coli. Four hours following inoculation of adult and newborn mice with a quantity of organisms 2 logs below the LD100, all animals were neutropenic. After 24 h, adults had recovered from the neutropenia but neonates had not (p < 0.001). Accelerated neutrophil production was evident in the infected adults, and correlated with the appearance of granulocyte colony-stimulating factor (G-CSF) transcripts in the liver, spleen, and lung, and interleukin-6 (IL-6) transcripts in the spleen and lung. An increase in neutrophil production was not observed in the neonates, and none of their organs tested had transcripts for either G-CSF or IL-6, but they did have transcripts for cytokines not involved in granulocytopoiesis; macrophage colony-stimulating factor and its receptor (c-fms). We speculate that the failure to increase neutrophil production in infected neonatal mice is the result of failure to increase production of relevant cytokines.

Animals↗

Reference intervals for 21 clinical chemistry analytes in arterial and venous umbilical cord blood.

Reference intervals were determined for 21 clinical chemistry analytes in umbilical cord arterial and venous blood from healthy term infants. Nonparametric analysis (rank number) was used to determine the central 95% reference interval. No significant differences were observed between male and female infants. Reference intervals for glucose, urea, creatinine, urate, phosphate, calcium, albumin, total protein, cholesterol, triglycerides, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatine kinase, lactate dehydrogenase, gamma-glutamyltransferase, and magnesium all were significantly different from adult values.

Blood Chemical Analysis↗

Characterization of endothelin receptors in the inner medullary collecting duct of the rat.

Endothelins may be important regulators of renal inner medullary collecting duct (IMCD) function. These peptides are secreted in large amounts by IMCD cells and can, in turn, potently inhibit sodium and water transport systems in the IMCD. This study characterized endothelin (ET) receptors in the IMCD in order to gain insight into this unique renal autocrine system. Radioligand binding studies with 125I-ET-1 yielded a B(max) of 205.7 fmol/mg and a KD of 218 pM for ET-1. Similar studies with 125I-ET-3 yielded two populations of receptors for ET-3, one with a KD of 50 pM and one with a KD of 920 pM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of IMCD cells covalently labeling with 125I-ET-1 yielded two bands, one at 97 kDa with affinities of ET-1 greater than ET-2 much greater than ET-3 and one at 47 kDa with affinities ET-1 greater than or equal to ET-2 = ET-3. Reverse transcription and polymerase chain reaction revealed the presence of both endothelin receptor types A and B. These data indicate that IMCD cells have high affinity, high density receptors for endothelin and express both known types of endothelin receptor.

Animals↗

Identification of the gene for the developmentally expressed 70 kDa heat-shock protein (P70) of mouse spermatogenic cells.

Mouse spermatogenic cells synthesize a 70-kDa protein (P70) closely related to the major heat-shock protein (hsp70) of mammalian cells (R. L. Allen, D. A. O'Brien, and E. M. Eddy, Mol. Cell. Biol. 8, 828-832, 1988). Expression of P70 is developmentally regulated while hsp70 is induced in response to stress, suggesting that P70 is the product of a unique member of the Hsp70 multigene family transcribed in spermatogenic cells. A strong candidate for this gene was the Hsp70.2 gene (Z. F. Zakeri, D. J. Wolgemuth, and C. R. Hunt, Mol. Cell. Biol. 8, 2925-2932, 1988). A DNA segment from the 5' region of Hsp70.2 hybridized to a 2.7-kb transcript with a temporal pattern of expression in mouse spermatogenic cells similar to the P70 protein. We used a polyclonal antiserum generated against a synthetic peptide predicted from the Hsp70.2 sequence to characterize its protein product and to isolate cDNA clones from a pachytene spermatocyte expression library. The antiserum reacted specifically with meiotic and postmeiotic spermatogenic cells on sections of mouse testis. It recognized the P70 protein on Western blots of two-dimensional gels and did not bind to other heat-shock proteins of spermatogenic or somatic cells. The cDNAs hybridized to a 2.7-kb mRNA that was abundant in unstressed pachytene spermatocytes and round spermatids but was not detected in other cell types. Two cDNAs were sequenced and found to be 99% homologous to the 3' end of the Hsp70.2 gene. These data strongly supported the hypothesis that P70 is the expressed product of the Hsp70.2 gene in mouse spermatogenic cells.

Amino Acid Sequence↗

Effect of smoking on fibronectin production by human amnion and placenta.

Fibronectin production from amnion and placental tissues was evaluated in pregnant woman smokers and nonsmokers in order to examine if there were alterations of fibronectin metabolism in intrauterine tissues. In both amnion and placental tissues, cycloheximide inhibited the fibronectin output indicating that it was being synthesized. Mean fibronectin output by amnion in pregnant woman smokers was significantly lower than that in pregnant woman nonsmokers. In contrast, in the placenta from pregnant woman smokers, the output was significantly higher than that in pregnant woman nonsmokers. The present observations indicate that smoking alters an important biochemical constituent in amnion and placenta, possible leading to some complications.

Amnion↗

Elimination of fibrinogen interference in a dye-binding method for iron.

Serum iron concentrations greater than 90 mumol/L were measured in samples from hemodialysis patients by a Ferrozine dye-binding method. Reanalysis by coulometry showed these results to be spuriously high. Turbidity resulting from precipitation of fibrinogen was identified as the source of the interference. High concentrations of heparin enhanced the effect. We conclude that the persistence of fibrinogen in serum samples from patients treated with anticoagulants is a potential interference in Ferrozine dye-binding methods performed without prior deproteinization. Adding guanidine.HCl to the acid buffer reagent is a simple way to eliminate this interference.

Coloring Agents↗

Aggregated bovine IgG inhibits mannose receptor expression of murine bone marrow-derived macrophages via activation.

We previously described the presence of an inhibitory protein contained in the 20 to 40% (NH4)2SO4 precipitable fraction of FCS that down-regulates expression of mannose receptors on bone marrow-derived macrophages. We now identify aggregated bovine IgG as the main inhibitory component. Heat-aggregated bovine IgG was capable of down-regulating expression of the macrophage mannose receptor in a dose-dependent manner without inducing changes in ligand affinity whereas neither F(ab')2 fragments nor nonaggregated IgG displayed any inhibitory effect. Depleting of IgG from heat inactivated FCS by protein G affinity chromatography completely removes the inhibitory activity. Moreover, readdition of the Ig eluate from the protein G chromatography column restored inhibition in a dose-dependent manner. Macrophages were able to clear exogenously added aggregated bovine IgG, thus leading to loss of inhibitory activity in macrophage-conditioned media as compared to sham-conditioned media containing aggregated IgG. These results indicate that aggregated IgG down-regulates mannose receptor expression by macrophage activation via interaction with Fc-gamma R.

Animals↗

Monomeric IgG2a promotes maturation of bone-marrow macrophages and expression of the mannose receptor.

The macrophage mannose receptor, a 172-kDa lineage-specific glycoprotein, partakes in nonopsonin-mediated phagocytosis by recognition of terminal mannose residues on targeted particles. Because appearance of the receptor progresses with monocyte/macrophage differentiation, its expression is indicative of the maturational state of the cell. Monomeric IgG2a and IgG2b up-regulate mannose-receptor surface expression and biosynthesis by murine bone-marrow macrophage precursors as much as 7- to 12-fold in a dose-dependent manner. IgG2a accelerates macrophage mannose-receptor expression by several days during in vitro bone-marrow differentiation; however, treated and control cells ultimately express equivalent levels of receptor. Moreover, the effect is independent of cell cycle or ambient levels of colony-stimulating factor 1. The coinduction of another maturation-dependent lineage-specific antigen, F4/80, and the fact that macrophage precursors respond to IgG2a only within the first day of culture, indicate that the targeted cell is an early myelomonocytic precursor, responsive only during a short, early developmental window. The effect is specific for immunoglobulin molecules of the IgG2a and IgG2b subclasses and probably involves an Fc gamma-receptor signal-transduction pathway but not macrophage priming or activation. Most importantly, a paracrine mechanism of immunoglobulin-mediated bone-marrow macrophage differentiation is suggested by experiments in which basal levels of mannose-receptor expression are reduced by continual removal of B-cell-generated IgG from marrow cultures. Thus, IgG2a and IgG2b prompt mannose-receptor synthesis and bone-marrow macrophage differentiation and may, therefore, play a role in the regulation of macrophage differentiation in host defense.

Animals↗

1,25-Dihydroxyvitamin D3 induces monocytic differentiation of the PLB-985 leukemic line and promotes c-fgr mRNA expression.

1,25-dihydroxyvitamin D3 [1,25(OH)2D] is known to stimulate maturation of the human promyelocytic line HL-60 and murine bone marrow precursor cells along a monocyte/macrophage pathway. In this study, the steroid's effects on the PLB-985 leukemic line were examined. We found that 1,25(OH)2D induces monocytic differentiation of PLB-985 as manifested by morphological appearance, histochemical staining, and changes in cell surface antigen expression. Additionally, there was acquisition of functional monocyte characteristics including phagocytic activity and superoxide anion production via the respiratory burst pathway. Steady state levels of mRNA derived from the leukocyte-specific proto-oncogene c-fgr were also increased by the steroid. Thus, 1,25(OH2)D effectively differentiates PLB-985 cells along a monocytic pathway, providing a useful model of macrophage differentiation.

Calcitriol↗

1,25-Dihydroxyvitamin D3 modulates colony-stimulating factor-1 receptor binding by murine bone marrow macrophage precursors.

1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] has been shown to be a potent agent for monocyte/macrophage differentiation in leukemic cell lines. This study examines the differentiational effects of 1,25-(OH)2D3 in authentic murine bone marrow immature uncommitted precursor cells collected by pretreatment of murine bone marrow with 5-fluorouracil (5-FU). Early precursor cells, collected 1 day after 5-FU treatment, are characterized by low expression of the colony-stimulating factor-1. (CSF-1) receptor and dependence on both CSF-1 and interleukin-1 for growth. Intermediate precursors were collected 5 days after 5-FU treatment and required either CSF-1 or interleukin-3 for growth. Intermediate precursors expressed relatively higher levels of the CSF-1 receptor. Addition of 1,25-(OH)2D3 to these populations inhibited proliferation, as measured by [3H]thymidine incorporation and soft agar colony assay. Furthermore, 1,25-(OH)2D3 caused a more rapid appearance of the CSF-1 receptor in early precursor cells in a dose-dependent metabolite-specific manner. Conversely, CSF-1 receptor expression was decreased in intermediate precursors treated with the steroid. This decrease in receptor expression was also dose dependent and metabolite specific. These findings demonstrate that 1,25-(OH)2D3 1) targets to authentic bone marrow macrophages at various stages of differentiation and 2) modulates expression of the CSF-1 receptor, a protein which, in turn, regulates monocytic maturation.

Animals↗