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Lectin histochemical studies were performed on formalin-fixed, frozen, and paraffin-embedded tissue sections from 19 patients with glucosylceramide lipidosis (i.e., Gaucher disease). Eleven different lectins were used to identify the specific carbohydrate residues in the undegraded stored compounds in the cytoplasm of Gaucher cells. In all cases studied, Gaucher cells stained with Concanavalia ensiformis agglutinin, Datura stramonium agglutinin, Lens culinaris, Ricinus communis agglutinin-I, and wheat germ agglutinin. These results demonstrated common carbohydrate residues in the undegraded material stored within Gaucher cells and indicated the presence of fucosylated N-linked complex oligosaccharides, and glycans containing N-acetyllactosamine repeating sequences, as well as nonreducing terminal beta-galactosyl and sialyl residues. In order to confirm these findings using biochemical methods, livers and spleens from Gaucher patients and controls, and from a patient with Niemann-Pick disease type C (included for comparison) were digested with Pronase and the resulting glycopeptides separated by gel filtration into fractions with high and low molecular weight. In the high-molecular-weight fractions from livers of Gaucher patients, the levels of sugars corresponding to N-linked glycans, as measured by gas-liquid chromatography, were elevated over those in controls. In the high-molecular-weight fractions from spleens, the levels of the same sugars were elevated in both Gaucher and Niemann-Pick type C patients. Digestion of the glycopeptides with endo-beta-galactosidase, which specifically cleaves polylactosaminoglycans, showed the presence of material containing N-acetyllactosamine repeating units in Gaucher liver glycopeptide fractions, but not in control and Niemann-Pick type C derived glycopeptide fractions. Our histochemical and biochemical studies demonstrated that in addition to glucosylceramide, affected tissues of patients with Gaucher disease accumulate glycoproteins. This accumulation could not have been predicted on the basis of the primary enzymatic defect.

To classify cases of non-Hodgkin's lymphoma in terms of expected clinical behaviour and survival, kinetic parameters measured by cytophotometry were assessed in 62 patients between 1978 and 1987. The influence of the number of cells with increased DNA content (more than 2N) on survival was evaluated. Analyses were carried out on the small samples obtained by needle aspiration biopsy of lymph nodes before treatment, using microdensitometry and Feulgen staining. Patients whose lymphomas contained less than 6% of cells with increased DNA content had a mean survival of 81.3 months and those whose lymphomas contained 6% or more of such cells had a mean survival of 18.5 months. A significant difference in survival using the same criteria was also noticed for patients with both low grade lymphomas and those with intermediate and high grade lymphomas. It is concluded that cytophotometric analysis of lymph node aspirates is of prognostic value in the initial assessment of non-Hodgkin's lymphoma.

We describe a case of "sinus histiocytosis with massive lymphadenopathy" (SHML) studied by immunohistochemical, cytogenetic and molecular analysis. The immunophenotyping showed that the lymph node histiocytes were strongly positive for the S-100 protein and MoAb LeuM3, OKM5, KP1 and DRC-1; a portion of these cells was also positive for OKT6 and Leu3A, suggesting a possible relationship with the veiled cells, which represent an intermediate step in the pathway from the Langerhans cell to the interdigitating reticulum cell. Cytogenetic analysis showed a normal prevalent clone and a small hypodiploid clone and the molecular study showed no detectable involvement of the c-fms proto-oncogene, which is related to monocyte/macrophages. Unfortunately all these data do not seem sufficient to define the benign or neoplastic nature of the disease. Further investigations, immunophenotypical, cytogenetic and molecular, are needed to elucidate the pathogenesis of the disease, especially for more aggressive cases or for cases with unfavorable evolution.

In the literature two elements, beryllium and zirconium, have been described as inducing identical tissular lesions consisting of granulomas, which some authors have related to an immunological process. Intracellular lesions induced by beryllium sulphate have been studied previously. These lesions were essentially characterized by intranuclear inclusions rich in beryllium. In this work we have studied intracellular concentration sites of zirconium after injection of low doses of zirconium sulphate. Results show that the intracellular concentration sites are very different between both elements, as zirconium is uniquely localized in the lysosomes of the lymph node macrophages where it is associated with phosphorus.

The present study was undertaken to evaluate the nuclear DNA content of esophageal cancer cells consequent on the process of growth and progression of the cancer. In experimental animal studies, 47 esophageal cancers were induced in male Wistar rats by oral administration of N-amyl-N-methylnitrosamine (AMN) and were analysed in the study of ploidy patterns. The study was also carried out to determine the DNA content in the ploidy patterns in man. Primary tumors associated 421 metastatic lymph nodes in the 62 patients with thoracic esophageal cancer were subjected for the study of ploidy patterns. The nuclear DNA content was determined by means of flow cytometry. In the study of the experimentally-induced esophageal cancer in rats, aneuploidy was found in 18% at a depth of submucosa, 30% at proprial muscle, 59% at adventitia, and in 50% at a depth of neighboring structures, respectively. Clinically in man, the incidence of DNA diploidy and aneuploidy in the 62 primary cancers was 56% and 44%, respectively. In the 421 metastatic lymph nodes, diploid was found in 73% and aneuploid in 11%, while the combination of diploid and aneuploid was observed in 16%. Difference in the DNA index (DI) between the primary cancers and metastatic lymph nodes was found in 29 cases (46.8%), and the difference increased with progression of the cancer. Two hundred and ninety seven metastatic lymph nodes of 29 cases were subdivided into 4 groups based on the extent of the cancerous nests, and the DI value was found to be increased in proportion to the extension. With the results, the DI value of esophageal cancer appeared to be changed dependently by the variation of cell populations in the cancer or in the DNA content of the cancer cells.

A combination of beta-galactosidase enzyme and the immunogold/silver staining method was studied for evaluation of double-staining experiments. Applications are shown for immunohistochemical double staining using two monoclonal antibodies and for combined immunohistochemistry and DNA in situ hybridization in one tissue section. The following advantages for the present double-staining method were evaluated: superior sensitivity of the immunogold/silver staining method for at least one epitope, which also allows detection of biotinylated DNA probes. The structure of the indolyl precipitate after revelation of beta-galactosidase activity did not show a concealing effect during a sequential double-staining method, as compared with the visualization of peroxidase with diaminobenzidine. These factors, and the sharply contrasting colored reaction products of beta-galactosidase (blue-green) and the immunogold/silver staining method including silver enhancement (brown-black), allow clear distinction of mixed-stained cell constituents.

A study has been conducted of the cellular DNA contents in the primary and metastatic lesions of 30 cases of lung-metastasized stomach cancers, in which each DNA Index (DI) was calculated and analysed comparatively. As a consequence, the rate of the diploid type in the nodular lung metastases was found to be higher than the rate of the others. Further, the DI's of 22 out of 30 (73%) metastatic lymph nodes corresponded with those of the primary lesions. Four lung metastatic lesions out of five differed from their primary lesions, but all five lesions simulated their metastatic lymph nodes. Thus understanding of the properties of the metastatic lymph nodes, rather than their primary lesions, would seen to be helpful in planning the therapy to combat recurrent cancers.

We present a new method for detection of micrometastases to axillary lymph nodes and estrogen receptor determination. Cellular suspensions from primary infiltrating ductal breast carcinoma or level I axillary lymph nodes of patients who underwent mastectomies were obtained, by loosely grinding fresh tumors or lymph nodes through a grid and then transferring the matrix to a slide using cytocentrifugation. Tumor samples were analyzed for estrogen receptor status using an immunocytochemical kit and compared with the dextran-coated charcoal method. Thirty-eight of 48 correlated (20 were estrogen positive, and 18 were estrogen negative). Seven of 46 were estrogen positive while results from the dextran-coated charcoal method were estrogen negative. One of 46 was estrogen negative, while the results from the dextran-coated charcoal method were estrogen positive. Lymph node slide preparations were stained to detect tumor cells using antikeratin monoclonal antibodies. Three of 8 node-negative patients were found to have micrometastases. Four of 15 node-positive patients had additional nodes with tumor. Our method combines the advantages of serial sectioning and immunohistochemical staining.

Double-labeling immunofluorescence of guinea pig tracheobronchial lymph nodes revealed complete coincidence of SP and CGRP immunoreactivities in perivascular nerves and axons of the medullary lymphatic tissue. Additional dynorphin A or cholecystokinin immunoreactivity was seen only in some of the medullary fibers. Ultrastructurally, all SP-immunoreactive axons were unmyelinated and displayed vesicle-containing varicosities. Retrograde neuronal tracing combined with immunohistochemistry revealed a sensory origin from dorsal root ganglia of SP/CGRP-immunoreactive fibers ramifying within paratracheal lymph nodes, and an additional neuronal population being devoid of SP/CGRP immunoreactivity. The findings provide evidence for several types of sensory nerve fibers innervating lymph nodes.

The immunophenotypic properties of the abnormal cells in routine specimens from 16 cases of Langerhans cell histiocytosis (LCH) were examined. In five cases, cryostat sections were also available. The abnormal cells expressed a similar phenotype and were positive for HLA-DR, S-100 protein, peanut agglutinin (PNA), CD1a, CD4 and several macrophage-associated markers, including CD11c, CDw32 and CD68 (the latter detectable in routine sections with antibody KP1). Staining with CD14, CD35 (C3b receptor), and CD11b (C3bi receptor) was negative with the exception of one of the cases in which a proportion of the cells showed faint positivity with CD11b. Staining for pan-T-cell (CD2, CD3, CD5) and pan-B-cell (CD19, CD22) antigens was negative in all lesions. It is concluded that LCH expresses a characteristic phenotype with some heterogeneity with regard to macrophage markers and that immunohistochemical methods in cryostat sections and routine specimens form a useful supplement to other techniques for the diagnosis of this condition.

This paper reports the nature of abnormally expressed Forssman (F) antigen in the lymph node cells of MRL/MpJ-lpr/lpr, autoimmune mice, and also reports its autoantibody in sera. By acetylation study of the F antigen with [14C]acetic anhydride, we concluded that the F antigen was not a glycolipid but a glycoprotein. Several bands of F-active glycoproteins were identified on a nitrocellulose sheet after purification by an anti-F antibody affinity column. Hemolysis of SRBC by some sera from MRL/MpJ/lpr/lpr was inhibited by purified F glycoprotein and also by F glycolipid. The antibody in the serum, however, seemed to be more specific for F glycoproteins than F glycolipid, but the opposite was the case for rabbit anti-F glycolipid antibody. No significant difference of the SRBC hemolysis levels was observed between the sera from MRL/MpJ-lpr/lpr and its congenic MRL/MpJ-+/+ mice.

The Authors analyse the functional areas (cortical, paracortical, germinal center, medullary, histiocytosis of the sinuses) of regional lymph nodes in women having uterine carcinoma and previously treated with Thymostimuline, and who were operated on respectively 8 and 14 days after inoculation. They also evaluate the degree of significance when compared with a control group treated locally with a physiological solution. The number of lymph nodes examined is at least 5, and the stereological analysis was carried out with the use of a grid: 200 points, actual area 200 cm2, effective area at the level of the preparation 0.32 mm2 (250 X). The Authors examine the results obtained after 8 days' action by Thymostimuline, and, after 14 days, the difference between the 8-day-after and the 14-day-after administration results, and those between the cervical and the endometrial. 4 women with uterine cancer (Stages I and II) and treated with the same doses of physiological saline solution were used as controls.

Using Falck fluorescent technique in combination with microspectrofluorometry the measurement of catecholamine and 5-hydroxytriptamine level was performed in popliteal lymph nodes of normal dogs and 2 weeks after deafferentation. Maximal catecholamine and 5-hydroxytriptamine content was observed in adrenergic nerve fibers, elements of trabecular-elastic complex and monoaminocytes of medullary substance of intact animals. Biogenic amine level decreased deafferented limb lymph nodes. Monoamine imbalance according to divergent type was registered in a colateral limb, i. e. 5-hydroxytriptamine level was increased and catecholamine level was decreased.

Zn, Cu, Cd and Pb contents in liver, spleen and lymph nodes of 30 cows with lymphatic leukemia were determined using polarographic method. In leukemic tissues more or less prominent increase of mentioned metals was observed. Results of the examinations would indicate a partical disappearance of natural antagonism between particular elements in neoplastic tissues. Metals accumulation in leukemic tissue is prabably a secondary phenomenon in relation to virus infection. Alterated by virus physicochemical cell properties promote not only accumulation bat also perversion of heavy metals to tissues.

Histiocytic (HRC) and interdigitating (IDC) reticulum cells were obtained from human lymphoid tissue (lymph nodes, tonsils), suspended in cell culture medium and cultured. The suspended cells, which were identified by morphological and enzyme histochemical criteria, were studied for surface binding properties for Fc-receptors using erythrocytes coated with IgG. HRC show strong surface binding, IDC, somewhat weaker, but definitely positive surface binding. Both types of reticulum cells possess Fc-receptors; on IDC, however, they are fewer in number and less dense. In connection with recently discovered Fc-receptors on Langerhans cells of the epidermis, functional relationships between blood monocytes, Langerhans cells in skin and IDC in lymphoid tissue are discussed.

Effects of pregnancy and lactation on the spleen and the mesenteric, iliac, axillary and popliteal lymph nodes in the mouse were morphologically examined by means of qualitative and quantitative methods. During pregnancy the spleen weight increases to reach a peak at 15 days of pregnancy. After parturition the weight decreases gradually until 20 days postpartum when it has returned to almost normal. Both the red and white pulps increase in volume during pregnancy. After parturition, the volume of the white pulp rapidly returns to normal. The volume of the red pulp remains greater in lactating females than in control females. In non-lactating females the red pulp decreases rapidly in volume after parturition. In the spleen, plasma cells increase in number during pregnancy, and often contain Russell bodies. They are accumulated in large numbers particularly in the marginal zone and periarterial area. After parturition plasma cells decrease rapidly in number. The weight of the mesenteric lymph node undergoes a slight but a significant increase during pregnancy, and decreases in both the lactating and non-lactating groups after parturition. During pregnancy the medulla shows an increase in volume, and the medullary cords are filled with numerous plasma cells. At 10--15 days of pregnancy, plasma cells also occur in groups in the paracortex. They decrease rapidly after parturition. During pregnancy plasma cells are also markedly increased in iliac lymph nodes, but not in axillary and popliteal lymph nodes. The findings were discussed particularly in relation to the immune response of the peripheral lymphatic tissue in pregnancy.

Plasma membranes from tumor cells in enzootic bovine leukosis and from normal bovine lymphoid cells of different sources have been isolated and characterized. The glycoprotein composition of the different membranes has been studied by SDS-PAGE and by analysis of the carbohydrate composition as well as by lectin binding of glycoprotein fractions obtained by phenol treatment of the membranes. No differences in the electrophoretical glycoprotein pattern could be detected comparing (nonmalignant) cells from persistent lymphocytosis and tumour cells, suggesting that malignancy is not associated with the gain or loss of a major glycoprotein. A 151.5 kD glycoprotein, present in membranes from normal lymph node lymphocytes, seems to be absent in the membranes of peripheral blood lymphocytes as well as of tumour cells. The loss of this glycoprotein might thus be associated with a loss of sessility of bovine lymphoid cells. The carbohydrate analysis and lectin binding of extracted glycoproteins revealed a decreased fucosylation accompanied by an increased exposure of galactose residues as well as a loss or a decreased complexity of N-glycosidically bound oligosaccharides in the tumour cell glycoproteins compared with those of normal cells. These findings are discussed with regard to disturbed growth regulation of leukosis tumour cells.

In a series of approximately 80,000 lymph nodes, amyloid deposition was found in 18; 12 of those nodes were selected, on the basis of availability of specimens, for investigation by immunohistochemical typing to identify the protein of origin and by correlation with morphologic criteria and clinical information. Four patterns of amyloid deposition were identified: lymph node vessel involvement, follicular deposition, diffuse deposition, and a combination of follicular and diffuse deposition. All cases were classified immunohistochemically with the amyloid type-specific antisera anti-AA, anti-A lambda, anti-A kappa, anti-ASc1, and anti-AF. Immunoglobulin-derived protein (AL) in lymph nodes was found in every case of isolated amyloidosis, lymphoplasmacytic/lymphoplasmacytoid immunocytoma, plasmacytoma, and idiopathic amyloidosis. Among the cases of AL amyloidosis were nine of A lambda and one of the A kappa type. AA protein was present in two cases of reactive systemic amyloidosis. There was no useful morphologic correlation with the immunohistochemically identified amyloid types.

Thirty-five patients with dermatopathic lymphadenopathy, 12 with (9 patients and 3 follow-up) and 23 without associated mycosis fungoides, and 7 controls with 26 lymph nodes were studied by light microscopic examination and immunostaining for S-100 protein. Fourteen of these cases were examined ultrastructurally. The group of cases studied included lymph nodes obtained at postmortem examination from three patients with mycosis fungoides and evidence of nodal involvement by light microscopic examination. Lymph nodes from patients with dermatopathic lymphadenopathy and paracortical hyperplasia (but no effacement of architecture by recognizable mycosis fungoides cells by light microscopic examination) associated with biopsy-proven mycosis fungoides showed well-defined, diffusely distributed sheets of S-100-positive cells. Cases of dermatopathic lymphadenopathy unassociated with a skin lymphoma (23 cases), with the exception of 3 cases (12.5%), only showed scattered S-100-positive cells, a similar pattern to that noted in normal or reactive lymph nodes from the control cases and reported literature. In the three cases with diffuse sheets without associated mycosis fungoides, the intensity of S-100 staining was the same in germinal centers as in parafollicular areas, while in cases associated with mycosis fungoides, the staining was predominantly in parafollicular zones. Ultrastructural immunolabeling for S-100 protein also revealed different patterns in both subsets of patients. Interestingly, in cases in which the lymph nodes were identified by light microscopic examination to be replaced by mycosis fungoides, the sheets of S-100-positive cells disappeared. The corresponding ultrastructural evaluation showed cellular aggregates with features of T-cells. A combination of S-100 immunocytochemistry and morphologic ultrastructural assessment can be of help in evaluation of lymphadenopathy in patients with mycosis fungoides. Although this study indicates that the finding of sheets of S-100-positive cells in patients with dermatopathic lymphadenopathy is not always associated with mycosis fungoides, the identification of diffuse sheets of S-100-positive cells, especially with parafollicular distribution, in a patient without known T-cell lymphoma should probably be considered an indication to suggest a complete clinical evaluation to rule out the possibility of an undiagnosed mycosis fungoides. The finding of sheets of S-100-positive cells in patients with proven mycosis fungoides may be a morphologic indicator of impending extracutaneous dissemination.

Necropsy materials from 57 cases of generalized amyloidosis in rhesus monkeys were reviewed. Clinically, animals with the disease were characterized by cachexia with muscle wasting, recurrent diarrhea, and arthritis. Gross lesions included hepatomegaly, splenomegaly, chronic/active colitis, fibrous strictures of the cecocolic junction, osteoarthritis, and generalized muscle atrophy. Histologic examination revealed minimal to severe deposits of amyloid in the small intestine (100%), spleen (93%), large intestine (67%), liver (40%), lymph nodes (71%), stomach and/or adrenal gland (32%). More amyloid was deposited in the spleen, liver, and small intestine than in other organs. Shigella sp. were isolated from feces in 23% of the cases and 84% had histologic evidence of colitis. Other findings indicated that 100% of the animals had lung mites, 25% had strictures of the cecocolic junction, and 40% had osteoarthritis. Thirty percent of the cases occurred in animals 10 months to 5 years of age, 10% in ages 6 to 10 years, and 60% in animals greater than 10 years old.

In the course of a chronic experimental study of polyarthritis in rats, we have established a plastic embedding technique which allows the processing not only of soft tissues but also of tissues such as cartilage, intraarticular ligaments, and even undecalcified whole rat joints and provides broad survival of enzymes and antigens. Preservation of morphological details and the precision of antigen localization are superior to those seen in cryostat and paraffin sections. The method seems to have much diagnostic and investigative value, especially in arthritis research.

In vitro culturing of lymph node cells from a human non-Hodgkin lymphoma gave rise to several colonies of eosinophil-like cells. Eosinophil colonies originated from cells that during the first week of culture had a fibroblast appearance and were adherent to plastic. The tissue culture was sacrificed after 14 days. At that time each colony was formed by 20-50 cells with intracytoplasmic peroxidase-positive and eosinophilic granules. Cells comprising the colonies exhibited different degrees of differentiation. Some of the cells (26.6%) were mature eosinophils, the majority (66.8%) resembled eosinophil myelocytes, and some other (4.6%) had a fibroblast appearance. One or two multinucleated giant cells were often present in the center of most of the colonies. These cells contained up to 10 nuclei, which were arranged in a "ring form" or centrally located; giant cells with a single, central, large, multilobed nucleus were also observed. Cells belonging to other myelopoietic lines could not be identified in the tissue culture. Histological examination of the lymph node revealed extensive presence of eosinophils at various degrees of maturation but absence of other myelopoietic lines.

Two Chinese patients with sinus histiocytosis and massive lymphadenopathy are reported. The results of enzyme and immunohistochemical studies are presented.

A receptor for Helix pomatia lectin (HPL) occurs on 71.5% of peripheral blood lymphocytes. With double incubation techniques we could show that the population of HPL-positive lymphocytes is by far (greater than 90%) identical with the population of T lymphocytes, as determined by "classical" markers (E rosetting) and newly developed T cell markers like OKT3 or anti-human Lyt3 monoclonal antibodies. In formaldehyde-fixed lymph node sections we could demonstrate the homing area of HPL-positive lymphocytes, the paracortical regions. Conventional histological counterstaining was readily possible. We have employed HPL-peroxidase for differentiation of lymphocytes in benign and malignant infiltrates in human skin. In reactive cutaneous infiltrates, as well as in cutaneous lymphomas, HPL-peroxidase staining allows a clear differentiation of the involved lymphatic cells, and may serve as a tumor marker in certain T lymphomas, where we could demonstrate the lymphoma cells to carry the HPL-receptor.

Lymph node lesions of tuberculous cattle and swine gave, after disruption, centrifugation through 2.2 M sucrose, and ultrafiltration, a material that brings about, in vitro, modification of the tubercle bacilli or chromogenic mycobacteria into bacterial elements that are not acid fast, rapidly growing on nutritive agar supplemented with glycerol. The phenomenon is similar to that which the authors have previously described, using an inducing agent extracted from cultures of mycobacteria, called "endometallaxic conversion."

With the use of the cytochemical stain for horseradish peroxidase of Graham and Karnovsky (1966. J. Histochem. Cytochem.14:291), conjugates of horseradish peroxidase with ricin, wheat germ agglutinin, and phytohemagglutinin were employed for the morphologic demonstration of d-galactose (ricin), N-acetylglucosamine (wheat germ), and N-acetylgalactosamine (phytohemagglutinin) containing moieties on the surface of unfixed, or paraformaldehyde-fixed rat lymphoid cells. D-Galactose, or d-galactose containing disaccharides inhibited the interaction between ricin peroxidase and lymphoid cell surface; also, N-acetylglucosamine inhibited the wheat germ peroxidase-lymphoid cell interaction, but N-acetylgalactosamine failed to inhibit the reaction between phytohemagglutinin peroxidase and the surface of lymphoid cells.

The development of B16F10 liver metastases as related to its vascular organization was studied by morphological and functional methods, using the DNA-binding fluorochrome, Hoechst 33342. Experimental metastases were produced by intrasplenic injection of B16F10 melanoma cells, and the animals were sacrificed 3-7 days after tumor cell injection. The results show that early metastases are made up of avascular nodules of tumor cells, which subsequently developed lacunae which are not lined by endothelial cells and usually contain blood cells. A more developed metastasis seems to be made up of several nodules with or without lacunae. Between the nodules, vessels connected to blood circulation were usually observed. A functional study with a fluorescent dye showed that early metastases stained negatively, while more developed metastases showed a reticular fluorescent pattern, coincident with the intrametastatic vascular network and displaying a nodular image. In this case, the fluorescence displayed a gradient of intensity decreasing from the vessels towards the lacunae, where the tumor cells showed no fluorescence. In summary, our results suggest that B16 liver metastases have a nodular growth pattern, which is characterized by the formation, during their development, of lacunae not connected to the general circulation and that tumor cells have poor access to small molecules delivered from the blood.

The immunoperoxidase method was used to investigate the presence of intracytoplasmic lysozyme, alpha 1-antichymotrypsin (alpha 1-ACT), alpha 1-antitrypsin (alpha 1-AT), transferrin, and albumin in hyperplastic and inflamed human lymph nodes. Lysozyme was demonstrated in eosinophils, neutrophils, histiocytes, in epithelioid cells, mast cells, and some lining cells of lymph node sinuses. alpha 1-ACT was detectable in many, but not all histiocytes that stained for lysozyme, and in sinus histiocytes, epithelioid cells, and mast cells, but not in neutrophils or eosinophils. alpha 1-AT was demonstrable in mast cells, neutrophils, and some epithelioid cells, but not in histiocytes. Transferrin was found in mast cells, but not in any of the other cell types investigated. Albumin was detectable in a few epithelioid cells and giant cells of the Langhans type. Lysozyme, alpha 1-ACT, alpha 1-AT, transferrin, and albumin were never demonstrable in interdigitating reticulum cells, dendritic reticulum cells, or lymphoid cells.

Changes in the number of cells and the weight of various lymphoid organs of mice, such as the regional lymph node (right inguinal node), spleen, thymus, bone marrow, and peripheral blood, were followed after the subcutaneous injection of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K). For comparison, the changes after injection of various polyclonal lymphocyte activators (PLA) including various preparations of bacterial lipopolysaccharide (LPS) were concurrently studied. The number of cells of all of the lymphoid organs tested and that of nucleated cells in the peripheral blood decreased significantly within a few days after injection of CPS-K, and increased later. Above all, the increase in the number of cells and in the weight of the regional lymph node was most prominent (about 10 times larger than that of the normal control). Such a marked increase in the number of cells of the regional lymph node was not induced by the injection of any preparation of LPS or any other PLA tested. The initial decrease in the number of cells after CPS-K injection was most marked and long lasting in the thymus. Although LPS prepared by Westphal's method from Escherichia coli 055 or Salmonella enteritidis exhibited a stronger decreasing effect on the number of cells of the thymus, the effect of LPS prepared by Westphal's method from E. coli O111 or that by Boivin's method from E. coli O55 was similar to that of CPS-K. It is concluded therefore that CPS-K has the ability to decrease the number of cells of various lymphoid organs, especially that of the thymus, initially after injection, which is a property in common with LPS, and CPS-K has a unique ability to increase markedly the cells of various lymphoid organs, especially those of the regional lymph node, at later stages after injection. Considering that CPS-K exhibits a much stronger adjuvant effect on the antibody response than does LPS or other polyclonal lymphocyte activators, it is suggested that this extraordinarily potent activity of CPS-K in increasing the number of cells of the regional lymph node is closely related to its strong adjuvant action.

A routine measurement with two 200 mm-diameter phoswich detectors, placed one each side of the sternum, yielded an apparent lung content of 17 nCi 241Am for a subject. subject. Inspection of the data revealed that there were many more counts from the left side than from the right and that there was a greater number of Np L X rays observed, relative to the L gamma ray, than would be expected for a uniform lung distribution of 241Am. These observations suggested a shallow localised deposit in the left side of the chest. The subject's working history could not rule out an inhalation exposure, but he was known to have had a wound contaminated with Pu/Am in his left hand 16 yr previously. The wound had contained approx. 0.02-0.03 mu Ci 241Am initially, most of which was excised from the wound site after 50 days. Prompted by this knowledge we used a 50mm-diameter intrinsic Ge detector to examine the wound and relevant lymph node sites. The results demonstrated a localised deposit of 241Am in the axillary lymph nodes with approx. 1 nCi 241Am at the wound site. Subsequent measurements made with the phoswich detectors suggested that there was approx. 7 nCi 241Am in the lymph nodes with approx. 1 nCi 241Am in the liver, thereby accounting for most of the detectable 241Am contamination in the thorax.

The dust content and composition of lesions and hilar lymph nodes from the lungs of British coalworkers have been examined. Samples of macules, fibrotic nodules, and massive fibrosis (both peripheral and central sites) were dissected from 49 lungs. The highest mean dust concentrations (about 20%) were found in nodules and massive fibrosis. Overall there were no significant differences between the selected lesion types and their respective whole lung dust composition, although the central sites of massive fibrosis were found to contain on average a higher proportion of coal and a lower proportion of ash and its measured constituents, quartz and kaolin plus mica, than the edge of the lesion (p less than 0.001 for each component). There were striking differences between recovered lung and lymph node dusts. An examination of 180 specimens showed a mean quartz in lymph node dust of 20.3% compared with 6.1% in lung dust. As expected the proportion of quartz was greater in lymph nodes and lungs from men who had worked "low" rank (high ash) coal. By contrast with the corresponding figures for lung dusts, however, the mean proportion of quartz in nodes did not increase over the pathological range of pneumoconiotic lung disease. On average the proportions of kaolin and mica in lymph nodes reflect those found in lungs. The lymphotrophic nature of quartz was clearly shown although it was not possible to show an association between this clearance pathway and any particular type of lesion.

The concentration of lead in five lung segments and in regional lymph nodes was determined post mortem in ten subjects by atomic absorption technique. In each case there was a considerable variation in the concentration of lead in different lung segments, and, in general, the apical segment in the upper lobe tended to be more contaminated than segments in the lower lobes. With one exception, the lead levels in the regional lymph nodes exceeded those of the lung segments. Individual differences in the distribution of lead in the lungs may be due to differences both in exposure and physiological response. When comparative analyses are made, we suggest that samples be taken from the same and preferably the basal segments of the lung.

A new conjugate for the detection of anti-trinitrophenyl(TNP) antibodies was developed to study the localization pattern of specific antibody containing cells and extracellular antibody in vivo. By means of a bridging molecule, poly-L-lysine, nine TNP groups and six horseradish peroxidase (HRP) groups were joined in one conjugate. Thus a higher specificity (more hapten) was united with a higher staining intensity (more enzyme) in the same conjugate. This conjugate made possible the simultaneous detection of anti-TNP antibody containing cells and establishment of their class (immunoglobulin M (IgM) and IgG). It was also used for the demonstration of anti-TNP antibodies in tissues where a TNP-alkaline phosphate (AP) conjugate could not be used due to high AP (endogenous) background staining. Thus we demonstrated anti-TNP antibody containing cells in gut associated lymphoid tissue and anti-TNP-(TNP-ovalbumin) immune complexes in the glomeruli of the kidney. We suggest that poly-L-lysine is a suitable bridging molecule for the preparation of hapten-HRP conjugates.

The content of hydroxyproline was determined in cancerous and noncancerous tissues obtained from 40 patients with primary breast cancer, and investigated from a clinicopathologic viewpoint. The level of hydroxyproline in cancerous tissue was higher than that in normal mammary gland, but was lower than that of the tissue of mastopathy. In relation to the histologic type, the level was the highest in scirrhous carcinoma, followed by papillotubular and solid tubular carcinoma. With regard to lymph node metastasis, the hydroxyproline level was significantly higher in positive than in negative metastatic cases. The corresponding level tended to be higher in patients at stages II-IV than in those at stage I. There was no obvious relationship to tumor size. However, a significant, negative correlation was found between the hydroxyproline level and patient age. Thus, the content of hydroxyproline was found to be closely related to age and clinicopathologic factors such as the histologic type and degree of lymph node metastasis.

The tissue distribution of fibronectin (FN) was examined using a commercial anti-FN serum, the peroxidase-anti-peroxidase (PaP) technique, and paraffin sections of 22 lymph nodes affected by Hodgkin's disease. Vascular basement membranes and reticulin fibres are selectively stained and their structural changes in this pathological condition become readily visible. In contrast to the normal lymph node and to Hodgkin's disease with lymphocytic predominance, cases of mixed cellularity disease contain individual and focally grouped cells displaying intracytoplasmic FN. In nodular sclerosis these cells with fibroblast morphology are consistently numerous in the marginal zones of the cellular nodes. Strongly reacting mastocytes probably absorbed the applied antiserum non-immunologically. All the other cell types giving rise to the varying appearances of Hodgkin's lesions are consistently negative with respect to intracellular FN, including all forms of Hodgkin cells. We conclude that in Hodgkin's disease the immigration of FN-secreting fibroblasts is an integral part of the early sclerosing reaction, which in itself is a defence/repair mechanism closely related to scar formation.

Mononuclear cells in peripheral blood (PB) and cerebrospinal fluid (CSF) of seven patients and lymph nodes of three patients with clinically definite multiple sclerosis (MS) expressed antigens that reacted with monoclonal antibodies (MoAb) specific for HTLV-I p19 and p24 gag proteins. The labelled cells were visualized with immunoperoxidase staining and indirect immunofluorescence and identified at the ultrastructural level with immunogold technique. The frequency of these cells was low, ranging from 0.1% to less than 0.01% in blood. In CSF it was approximately 10 times lower. Cells reacting with anti-p19 Ab were found in all MS samples, whereas cells reacting with anti-p24 Ab were found in 3 out of 6 blood samples and in 3 out of 7 CSF samples. All lymph nodes (3/3) obtained from MS patients contained cells that reacted with anti-HTLV-I. p19 and p24 Ab. Cells reacting with the same AB were detected in blood of one out of 12 healthy controls. Stained cells were irregular, distinctly larger than lymphocytes, and had abundant cytoplasm, suggesting that they may be monocytes/macrophages. Immunogold particles were located in vacuole-like structures in the cytoplasm. The presence in MS patients of cells that react with HTLV-I Ab indicates that a human retroviral genome is being expressed, and suggests that a virus may be present. Our data support a role for a human retrovirus in multiple sclerosis.

Simultaneous cytophotometric and cytogenetic analyses of spontaneous mesenteric lymphomas from three female mice are reported. No statistically significant deviation from normal lymph node cells could be detected for any of the tumors with respect to nucleic acid content. However, all three tumors demonstrated trisomy as the major cytogenetic anomaly. Chromosome 7 or 8 was found to be the extra chromosome in two of the lymphomas while identification of chromosomes was not possible in the third.

Lymph node and spleen tissues involved in malignant lymphomas were analysed for iron, manganese, copper, zinc and magnesium by atomic absorption spectrophotometry. The levels of iron are found to be significantly lower in the case of Hodgkin's lymphoma compared with non-Hodgkin's lymphoma and normal lymph nodes. However, they are elevated in Hodgkin's lymphoma when compared with the normal value for spleen tissues. Magnesium is significantly higher in lymph nodes of non-Hodgkin's lymphoma compared with Hodgkin's lymphoma and normal values, but is not altered significantly in spleen tissues. The distribution of the other elements examined is not altered significantly in malignant lymphomas. The importance of the in situ levels of these elements to NMR imaging is discussed.

The concentrations of natural alpha-emitting isotopes of thorium (228Th, 230Th and 232Th) have been determined in 22 sets of human tissue samples obtained at autopsy from Grand Junction, CO and in 10 sets from Washington, DC. Tissues included lung, pulmonary lymph nodes, liver, kidney, bone, a few gonads, spleen and thyroid. Personal data on each individual's age, sex, smoking history and occupation were obtained whenever possible. The concentrations of 228Th, 230Th and 232Th were highest in lymph nodes for both populations with 2.6 and 5.1 pCi/kg of 228Th, 4.60 and 11.10 pCi/kg of 230Th, and 2.8 and 7.8 pCi/kg of 232Th in Washington, DC and Grand Junction, CO, respectively. The order of concentrations of all three isotopes in all other organs for both populations was as follows: (formula; see text) The data suggest that the non-mining residents who lived in the vicinity of uranium mine tailings do not have elevated 230Th concentrations in their lungs, when compared to the residents of Washington, DC who are not exposed to such tailings. However, 230Th concentration in bone of Grand Junction subjects was just significantly higher (at p less than 0.1) than that for Washington, DC subjects after suitable age adjustments. The data also suggest that 230Th is more available for accumulation in skeleton than would be supposed from its relative geochemical abundance.