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Biomedical subjects

A Hsu

Publications and source records attributed to A Hsu.

At least 91 records · Page 5Linked to original sources

HMB-45 staining of dysplastic nevi. Support for a spectrum of progression toward melanoma.

Dysplastic nevi are melanocytic tumors that occupy intermediate positions in the spectrum of melanocytic proliferations. Although they are invariably cured if completely excised, their biologic potential if left untreated is unknown. We examined a series of such lesions with HMB-45, a melanocyte-specific antibody, in order to explore protein expression within these borderline lesions. HMB-45 has previously been shown to label intraepidermal melanocytes within melanomas and within all nevi. Intradermal melanoma cells also label with HMB-45, but dermal nevus cells within common melanocytic nevi do not normally stain. In contrast, we found mild to moderate staining of nevus cells within the papillary dermis of dysplastic nevi and within residual nevus cells adjacent to malignant melanomas. In the same lesions, we demonstrated strong staining of intraepidermal melanocytes. Thus, dermal nevus cells within dysplastic nevi and within residual nevus cells adjacent to malignant melanomas are expressing low-level amounts of a protein expressed by melanoma cells, but not by dermal nevus cells within wholly benign melanocytic tumors. This lends support to the concept of these lesions as precursor lesions with undetermined biologic potential.

Antibodies, Neoplasm↗

Changes seen in lymph nodes draining the sites of large joint prostheses.

We report the histological findings seen in the lymph nodes draining the sites of large joint prostheses. Two patients underwent multiple prosthetic joint replacements. In one patient, the regional lymph nodes were enlarged during the revision of a total hip prosthesis, and a representative lymph node was resected. The other patient had undergone a pelvic lymph node dissection as part of a staging procedure for prostatic carcinoma. By light microscopy, the lymph nodes from both patients showed markedly dilated nodal sinuses filled with macrophages containing abundant eosinophilic, PAS-positive, granular material. Polarization microscopy revealed needle-like particles within the cytoplasm of the macrophages. We believe that the histological appearance of the lymph nodes represents a florid foreign body reaction to fragments of polyester or polyethylene derived from the articulating surfaces of the joint prostheses and transported to the regional lymph nodes via the lymphatic circulation. Sinus histiocytosis seen in the lymph nodes draining the sites of joint prostheses may resemble, and must be distinguished from, other conditions invoking a sinus pattern of lymphadenopathy, as well from benign and malignant diseases that involve the lymph nodes in a pattern mimicking sinus histiocytosis.

Adult↗

HMB-45 recognizes stimulated melanocytes.

A monoclonal antibody (HMB-45) was previously reported to bind to melanoma cells, and the junctional component of nevus cells, but not to normal adult melanocytes. We have tested HMB-45 binding in several conditions under which melanocyte stimulation might be expected in adults, i.e. 3 simple lentigines, 2 solar lentigines, 7 recent surgical scars (from re-excision of non-melanocytic tumors), 2 surgical scars from re-excisions of melanomas (after complete primary excisions), 9 hemangiomas from non-sun-exposed skin, 1 basal cell carcinoma, 1 acute ecchymosis, 1 keloid, and 1 dermatofibroma. Positive controls included 6 malignant melanomas and 1 fetal skin sample. Melanocytes were strongly positively stained overlying hemangiomas, within or near recent surgical scars of melanocytic and non-melanocytic tumor re-excisions, near basal cell carcinoma, and in fetal skin. Melanocytes either were not stained or were stained only focally for trace amounts in the normal skin near the new margins of the wide re-excision specimens for melanoma, i.e., at a distance from the scar, in the simple lentigines and in the fibrotic lesions. Thus, HMB-45 is staining an antigen which appears in adult melanocytes during stimulation and in fetal skin, as well as in melanomas. This stimulation is associated with conditions that would have increased vascularity, suggesting a melanocyte response to a plasma factor, or other endothelial cell derived factor. HMB-45 would not be a useful marker for residual melanoma cells in melanoma re-excision specimens.

Antibodies, Monoclonal↗

1,25-Dihydroxyvitamin D3 and rat vascular smooth muscle cell growth.

Recent studies from several laboratories have shown perturbations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] metabolism in hypertension. While these perturbations may exert their effect on blood pressure via their actions on calcium metabolism, it is possible that this vitamin D metabolite may have direct effects on vascular smooth muscle cell (VSMC) physiology. To examine this, we studied the effect of 1,25(OH)2D3 on VSMC growth and found that this substance suppressed VSMC [3H]thymidine uptake; furthermore, this vitamin D metabolite also suppressed the stimulatory effect of epidermal growth factor (EGF) on VSMC proliferation. The concomitant presence of this substance appeared to be required for its action on VSMC growth since cells pretreated with the vitamin D metabolite for up to 72 hours and then washed of the substance grew normally and responded to EGF. Studies were also done to determine if 1,25(OH)2D3 had any effect on the function of EGF receptors on VSMC. Experiments using Iodine-125-labeled EGF showed no differences in the binding of this ligand to VSMC, either untreated or treated with 1,25(OH)2D3, which indicates the effect of the vitamin D metabolite on VSMC growth (when exposed to EGF) was not mediated by an alteration of EGF receptor function. The results of these studies have implications for the pathogenesis of vascular diseases such as hypertension and atherosclerosis.

Animals↗

Penicillamine dermatopathy with lymphangiectases. A clinical, immunohistologic, and ultrastructural study.

The term penicillamine dermatopathy refers to the characteristic hemorrhagic skin lesions found in persons receiving long-term penicillamine therapy for either Wilson's disease or cystinuria. These lesions are thought to develop as a result of faulty collagen and elastin synthesis. We describe a patient with Wilson's disease who developed extensive penicillamine dermatopathy. In addition, histologic, immunochemical, and ultrastructural studies revealed multiple lymphangiectases with blood vessel to lymphatic anastomosis within these lesions, a finding not previously reported. The possible relationship to defective collagen and elastin formation are considered.

Adult↗

The effect of transforming growth factor-beta on mouse mesangial cell proliferation.

The factors which regulate mesangial cell growth are of importance in determining the cellular lesions of nephritis. In this communication, we report that transforming growth factor-beta (TGF-beta) exerts a suppressive effect on mesangial cell 3H-thymidine uptake. Furthermore, TGF-beta antagonizes the mitogenic effect of epidermal growth factor (EGF) on DNA synthesis. The concomitant presence of TGF-beta in the cell cultures is not required for its effect since cells pretreated with the substance and rinsed of it showed suppressed 3H-thymidine uptake and did not respond to EGF. The effect of TGF-beta can also be demonstrated on cells committed to proliferate by the prior addition of EGF. The results of the study have relevance to the mechanisms underlying the histologic lesions of immune-mediated nephritis.

Animals↗

Beta-endorphin amplifies the effect of interleukin-1 on mouse mesangial cell proliferation.

We report experiments that show that beta-endorphin, a neuropeptide, enhances the effect of interleukin-1 on mouse mesangial cell proliferation. Met-enkephalin, which comprises the first five amino acids of the beta-endorphin molecule, can also be shown to possess similar biologic activity. This action of beta-endorphin and met-enkephalin is mediated through naloxone-insensitive receptors on the mesangial cell, because naloxone treatment of the kidney cells does not abrogate this activity. The studies delineate a novel mechanism by which a neuropeptide may influence the development of immune-mediated cellular pathology.

Animals↗

Induction of mouse monocyte-macrophage replication by a mesangial cell-derived factor.

Conditioned media of cultured mouse mesangial cells (possessing microfilaments) were shown to contain a factor that stimulated splenic monocytes-macrophages and blood monocytes to replicate. Replicated cells were shown to express MAC-1 antigen as demonstrated by immunofluorescence with anti-MAC 1 and to possess Fc receptors as evidenced by their capacity to ingest sensitized erythrocytes. Preliminary characterization revealed the following characteristics: by Amicon ultrafiltration, fractions greater than 100,000 daltons were shown to have biologic activity; chromatofocusing of these active fractions revealed a peak of activity associated with fractions having pH 4; heating to 100 degrees C for 10 minutes abolished almost all activity, whereas trypsin treatment was without effect. The observations suggest a mechanism by which mesangial cells may modulate the proliferation of monocytes-macrophages that infiltrate the glomerulus in glomerulonephritis.

Animals↗

Isolation and characterization of Z-DNA binding proteins from wheat germ.

The preparation of a heterogeneous non-histone protein extract from wheat germ utilizing Br-poly(dG-dC).poly(dG-dC) (Z-DNA) affinity chromatography is described. The binding characteristics of antibodies against Z-DNA are used as a model system to define important criteria that the DNA binding behavior of a Z-DNA binding protein should display. We show that the wheat germ extract contains DNA binding proteins specific for left-handed Z-DNA by these criteria. The affinity of the proteins measured by competition experiments was approximately 10(5) greater for Br-poly(dG-dC).poly(dG-dC) (Z-DNA) than for poly(dG-dC).poly(dG-dC) (B-DNA). The affinity of the proteins for plasmid DNA increases with increasing negative superhelicity which is known to stabilize Z-DNA. The proteins are shown to compete with Z-DNA antibodies for binding to supercoiled plasmids. Finally, the affinity for two plasmids at a given superhelical density is greater for the plasmid containing an insert known to form Z-DNA than for a plasmid without the insert. The proteins exhibit a 2-3-fold greater affinity for stretches of (dC-dA)n.(dT-dG)n over stretches of (dG-dC)n.(dG-dC)n when both sequences are induced to form Z-DNA by supercoiling.

Animals↗

Evidence for an endothelial cell-derived factor which stimulates the growth of human peripheral blood mononuclear cells.

Evidence is provided which demonstrates that conditioned media of cultured endothelial cells derived from human umbilical veins contained a factor which stimulated peripheral blood mononuclear cell [3H]thymidine uptake. A dose-dependent response in peripheral blood mononuclear cell [3H]thymidine uptake was obtained when cells were incubated with increasing concentrations of supernatant of endothelial cell cultures. Studies on temporal kinetics demonstrated that stimulatory activity was evident when mononuclear cells had been incubated with endothelial cell supernatant for 120 hr or more. Preliminary characterization showed the growth immunoregulatory factor to have a molecular weight greater than 100,000 Da.

Cell Division↗

Evidence for a mouse mesangial cell-derived factor that stimulates lymphocyte proliferation.

The functions of the glomerular mesangium are served by at least two populations of cells--a cell bearing microfilaments that regulates blood flow, and a phagocytic cell bearing Ia determinants and Fc receptors. We provide evidence that mouse mesangial cells (bearing microfilaments) produce a factor(s) that stimulates spleen cell proliferation. The factor(s) appears to act via monocytes/macrophages, since its stimulatory activity is abrogated by prior depletion of the responding mononuclear cell population of monocytes/macrophages. Confirmation of its action on macrophages was documented by experiments that showed that medium from macrophages incubated with mesangial cell supernatant contained greater amounts of a factor that stimulated [3H]thymidine uptake by macrophage-depleted spleen cell populations. By the cothymocyte proliferation assay, it could be shown that mesangial cell supernatant induced splenic macrophage production of interleukin-1-like activity. Preliminary characterization reveals the factor to have a molecular weight greater than 100,000. Thus, a novel function is delineated for this mesangial cell type that appears capable of modulating the local immune response by providing an amplification signal.

Animals↗

Modulation of mouse mesangial cell proliferation by macrophage products.

Mesangial hypercellularity is usually found in many models of nephritis characterized by monocyte/macrophage infiltration of the glomerulus. In order to examine the mechanism mediating these events, an in vitro model was used to study the effects of macrophage products on mouse mesangial cells, cultured under conditions which would render them relatively quiescent. Under these conditions, macrophage supernatants stimulated the proliferation of the mesangial cells. The stimulatory effect could be shown to be due in part to enhancement of endogenous mesangial cell PGE production. This was demonstrated by experiments which showed that macrophage supernatants stimulated mesangial cell PGE production, that the stimulatory effect of macrophage products was abrogated by pretreatment of mesangial cells with indomethacin, and finally that exogenous PGE2 stimulated mesangial cell proliferation.

Animals↗

Mechanisms of suppression of mouse mesangial cell proliferation by macrophage supernatants.

The monocyte/macrophage has been identified as an effector cell infiltrating the glomerulus in human and experimental nephritis. To clarify the role of the macrophage in this context, an in vitro system was developed in which mouse mesangial cell cultures were maintained. Macrophage supernatants were obtained from peritoneal macrophages harvested from either resident or endotoxin-stimulated C57BL/6J male mice cultured for 24 hr. Incubation of mesangial cell cultures with macrophage supernatants resulted in depression of mesangial cell metabolism as indicated by incorporation of (3H)-thymidine; the effect was more marked when supernatants of endotoxin-treated mice were used. The molecular mechanisms by which suppression was obtained was clarified by experiments fractionating macrophage supernatants by G-100 column chromatography. By this means, two fractions were obtained with different molecular and physiologic properties. One fraction, a molecular size of 14,600 to 29,000 daltons, was shown to mediate the suppressive effect by stimulating endogenous mesangial cell PGE synthesis; additionally, a novel molecular species was identified, which was biologically active at higher concentrations of macrophage supernatants, had a larger molecular size (29,000 to 68,000 daltons), exerted its suppressive effect by an independent mechanism, and accounted for the inability of indomethacin pretreatment of mesangial cells to abrogate completely the suppressive effect of macrophage supernatant at higher concentrations.

Animals↗

Human mononuclear cell modulation of endothelial cell proliferation.

Endothelial cell proliferation is a histologic characteristic of several forms of nephritis characterized by infiltration of the glomerulus with mononuclear cells. To investigate the mechanism mediating this event, human endothelial cells isolated from umbilical veins and cultured in vitro were incubated with supernatants of cultured human mononuclear cells. Supernatants from mononuclear cells exerted a dose-dependent stimulatory effect on endothelial cell proliferation. The stimulatory effect of supernatant was almost entirely removed by prior depletion of mononuclear cells of monocytes by adherence, suggesting that a monocyte product was responsible for the activity. To investigate the nature of the ligand responsible, partially purified human interleukin I added to endothelial cell cultures was found to stimulate cellular proliferation.

Cell Adhesion↗

Interaction of alpha-dansylated peptide inhibitors with porcine pepsin: detection of complex formation by fluorescence energy transfer and chromatography and evidence for a two-step binding scheme.

Peptide inhibitors, specifically labeled at the alpha-amino terminus by dansylation, have been prepared by utilizing solid-phase peptide synthesis. Changes in fluorescence have been observed upon mixing these peptides with porcine pepsin that can be attributed to the formation of at least two complexes. Energy transfer between tryptophan residues of the protein and the dansyl group of the inhibitors has been detected by the unique excitation spectra generated. The kinetics of formation of the second complex can be correlated with inhibition of the catalytic activity of pepsin. Evidence for complex formation has also been obtained from gel filtration experiments using the fluorescent peptides.

Animals↗

Diminished synthesis of immunoglobulin by peripheral lymphocytes of patients with idiopathic membranous glomerulonephropathy.

Some studies of animal models of serum-sickness nephritis have shown that the lesions of membranous nephropathy develop in animals exhibiting a poor antibody response to the administered antigen (if given in constant amounts). It is postulated that patients with idiopathic membranous nephropathy may share a similar characteristic, namely, a diminished capacity to produce sufficient amounts of antibody. To test this hypothesis, we examined the ability of lymphocytes isolated from 11 patients with this disorder to produce immunoglobulin (Ig)G and IgM on stimulation with a polyclonal B-cell activator, pokeweed mitogen. The peripheral blood lymphocytes (2 x 10(6) cells) from 24 normal individuals had geometric mean production rates of 1,779 ng for IgG, and 2,940 ng for IgM after 7 d of culture in the presence of pokeweed mitogen. By contrast, under identical conditions, lymphocytes from the 11 patients with membranous nephropathy produced significantly lower quantities of both immunoglobulins, with geometric mean concentrations of 511 ng for IgG and 439 ng for IgM. When lymphocytes from patients with membranous nephropathy were co-cultured with normal lymphocytes, the production of immunoglobulin by normal lymphocytes was depressed by 22-82%, suggesting that a population of suppressor cells was responsible for this disturbance in B-cell function. By co-culturing normal lymphocytes with patient lymphocytes depleted of either T cells or monocytes, the suppressor cell was identified as a monocyte.

Adolescent↗

Comparison of chorismate mutase isozyme patterns in selected plants.

A wide variety of plants have been assayed to determine if they contain three isozymes of chorismate mutase (EC 5.4.99.5) as does alfalfa (Medicago sativa L.) or two isozymes, as does mung bean (Phaseolus aureus). The isozymes were separated by disc electrophoresis. All anthophyta with the exception of some closely related Leguminosae contained three isozymes of chorismate mutase. The one coniferophyta (a pine), and pterophyta (a fern) and one microphyllophyta (a Selaginella) assayed contained two isozymes of chorismate mutase. All plants assayed contained measurable chorismate mutase levels and at least two isozymes of chorismate mutase.

Journal Article↗