Ontogeny of B cells expressing IgM in embryonic and larval tissues of the American grass frog, Rana pipiens.

Affinity-purified, fluorochrome-tagged F(ab')(2) antibody fragments specific for heavy (mu) chains of Rana pipiens IgM were prepared from hyperimmune rabbit sera. By using two-color immunofluorescent procedures we observed that (1) the first cells expressing IgM, termed pre-B cells, lack detectable quantities of membrane or surface IgM but contain detectable quantities of cytoplasmic IgM (smu(-)/cmu(+)), (2) sIgM(+) B cells were the second type of IgM containing cell to appear in development, and (3) plasma cells, which contain copious quantities of cIgM, were the final phenotype to appear in the development of B cells expressing IgM. These cells were first observed in the pronephros of the developing urogenital system. Shortly after their appearance in the pronephros, cells in B lineages were observed in the liver. These observations (1) are consistent with recent studies of B lymphopoiesis in the aorta-gonad-mesonephros (AGM) region in endothermic vertebrates, including mice, (2) suggest that there are fundamental ontogenetic and phylogenetic similarities between cells and tissues of developing vertebrate immune systems, and (3) evoke questions concerning the possible function(s) of lymphocytes in developing anurans up to metamorphosis and beyond.

Immunohistochemical evidence that gonads and gonad-associated tissues are sites for enrichment with immunoglobulin-containing cells in adult chickens.

Immunohistochemical staining of tissue sections prepared from gonads and gonad-associated tissues obtained from adult chickens was performed in order to assess the possibility that these tissues may be sites of enrichment with IgM-containing cells in various B lineages. Evidence is presented which suggests that IgM-containing B lineage cells are present in 1) the ovarian stroma and subcapsular areas of the ovary and 2) the interstitium and subcapsular areas of the epididymis of the testes. These represent new sites reported for B lineage cells in adult chickens. Some questions relevant to the physiologic, ontogenetic, and phylogenetic implications of these observations relative to vertebrate hematolymphopoietic processes are included.

In vitro differentiation of B cells and myeloid cells from the early mouse embryo and its extraembryonic yolk sac.

The yolk sac is the first site of hematopoiesis during ontogeny. However, the source of early embryonic hematopoietic stem cells remains unresolved. Early studies have shown that cells obtained from day-8 and -9 extraembryonic yolk sacs can give rise to T cells and myeloid cells, whereas the embryo itself appears to lack such cells. Controversy remains as to whether it is the embryo itself or the extraembryonic yolk sac that contains the initial precursors capable of differentiating into B cells. This study used the approach of enriching hematopoietic stem cells by immunocytoadherence and studying cells isolated from within the embryo itself or from the yolk sac obtained at days 8 and 9 of mouse embryonic development. We report that on day 9, both yolk sac-derived and embryo-derived cells can give rise to B cells and myeloid cells in vitro. On day 8, however, cells isolated from the yolk sac but not from the embryo produce myeloid colonies in vitro; neither source of stem cells generates B cells. Our study suggests that myeloid precursors migrate from yolk sac to embryo earlier than has previously been reported but that the origin for B cell precursors remains to be determined.

Immunoglobulin-containing cells in chick embryo urogenital tissues: a new site for early B lineage cells in endothermic vertebrates.

We have employed histological and immunofluorescent staining procedures in order to characterize the distribution of mu + B lineage cells in tissue sections prepared from developing chicken embryo urogenital tissues (UGTs) between 14 and 21 days of incubation. B lineage cells were observed in tissue sections prepared from developing UGTs, especially the mesonephros and its associated tissue, throughout the sample period. The highest densities of mu + B lineage cells were observed in tissue sections prepared from 18 day embryos. The mu + UGT cells were distributed singly and in clusters in subcapsular regions and within the peritubular interstitium of the mesonephros. These observations (1) are consistent with those which suggest nonbursal site(s) for origin of cells in B lineage, (2) may help account for the varying effects of embryonic caudectomy performed between the second and third days of incubation and surgical bursectomy performed close to hatching, (3) may help provide new insights on the effects of sex hormones on B cell development, and (4) suggest fundamental ontogenetic and phylogenetic similarities between developing vertebrate immune systems.

Liver damage from flucloxacillin, cloxacillin and dicloxacillin.

We describe a case of severe cholestatic liver disease, which persisted for 7 years, and was probably induced by flucloxacillin. We also report a survey of 77 liver reactions which were probably or possibly induced by penicillinase-resistant penicillins and spontaneously reported to the Swedish Adverse Drug Reactions Advisory Committee. The reactions were usually cholestatic with a tendency to a protracted course. There is some evidence for an immunoallergic idiosyncratic reaction. The incidence of reported liver reactions was estimated from sales data to be 1.6-2.9 per million DDD (defined daily doses) or, for flucloxacillin 1:11,000-1:30,000 prescriptions. Female sex, age and high daily doses seemed to be associated with higher risk of liver reactions from flucloxacillin.

Immunochemical and immunohistochemical studies of cadmium associated proteins in Rana tadpoles.

Previous observations suggested that Rana tadpoles treated with aqueous cadmium (Cd) accumulate Cd in their liver and mesonephros. In order to study the response to Cd in these tissues we (a) exposed tadpoles in mid-limb bud stages to sublethal quantities of Cd, (b) isolated Cd-associated protein (CAP) from a liver cytosol fraction, (c) prepared a heterologous rabbit antiserum against glutaraldehyde-treated CAP (G-CAP), (d) used the rabbit anti-G-CAP antiserum in order to assess the tissue distribution of CAP in Cd-treated and untreated tadpoles, and (e) assessed species cross-reactivities of our anti-G-CAP with CAPs and metallothioneins (MTs) isolated from Cd-treated vertebrate liver cytosol fractions. We found that (a) CAP was present in higher quantities in liver cytosol obtained from Cd-treated tadpoles compared to liver cytosol obtained from untreated control tadpoles, (b) indirect immunofluorescent analysis revealed that CAP was localized in liver hepatocytes and kidney tubule epithelial cells in Cd-treated tadpoles, and (c) the anti-G-CAP crossreacted with rodent and fish CAP. These observations suggest that the developing liver and mesonephros are involved in responses to toxic metals and that our anti G-CAP antiserum may be used to gauge exposure to environmental Cd.

Effects of prolonged low-level cadmium exposure on the tadpole immune system.

Uninjected (Group I) and sheep erythrocyte (SRBC)-injected (Group II) Rana tadpoles were exposed to varying sublethal concentrations of cadmium (Cd) for 6 weeks. In order to assess possible effects on the tadpole immune system we determined pre-B, B mu, and plasma cell (PC mu) frequencies in liver and mesonephros of Group I larvae, and hemagglutinating antibody (HA) titers of Group II animals. Group I and Group II control animals were cultivated in water with no added Cd (0 ppm), while treatments were set at 0.1, 0.2, 0.4 and 0.8 ppm Cd. We found that (a) Cd appeared to stimulate a significant increase in the frequency of B mu cells in animals treated at 0.4 and 0.8 ppm, (b) certain treated Group II larvae contained significantly increased amounts of HA in their serum, while their serum protein concentrations were not significantly different, and (c) there was a dose-related increase in tissue Cd levels in treated Group II larvae. Our observations suggest that chronic low-level exposure to Cd may (a) result in a slight increases in the frequency of B mu cells in unimmunized animals, (b) increase immune responsiveness of immunized larvae, and (c) increase liver and mesonephros accumulations of Cd. Preliminary studies indicated that cytosolic Cd is associated with a protein which appears to be similar to mammalian metallothionein.

Liver involvement in Addison's disease.

Three patients admitted because of slightly increased serum aminotransferases were found to have Addison's disease. A review of 16 other patients with Addison's disease who had serum aminotransferase activity [aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT)] determined prior to treatment revealed one more patient with slightly increased aminotransferase activity that could not be explained by known causes of increased serum aminotransferase levels. In all four of these patients, the enzymes normalized within 1 wk of corticosteroid substitution treatment. Liver biopsy in one patient revealed discrete lymphocytic infiltrates in the portal zones. On the basis of these observations, we recommend that the possibility of Addison's disease should be considered in patients with obscure slight hypertransaminasemia.

Elevated aminotransferases and alkaline phosphatases in unstable diabetes mellitus without ketoacidosis or hypoglycemia.

Four patients with unstable diabetes mellitus and pronounced elevations of serum aminotransferases and alkaline phosphatases are reported. Thorough investigations revealed no cause for the abnormalities. The enzyme elevations were associated with hepatomegaly, and in some instances, abdominal pain and leg edema. Liver biopsies in all patients showed only marked accumulation of glycogen in the hepatocytes.

Relationship between gallbladder histopathology and ability to concentrate biliary lipids and bilirubin. A study on gallstone patients with functioning gallbladder.

Biliary lipids and bilirubin were measured in hepatic and gallbladder bile obtained at routine cholecystectomy in 35 gallstone patients. The gallbladders had opacified at cholecystography and the cystic ducts were patent at operation. The histologic changes in the gallbladder wall were evaluated by an independent pathologist. Increasing abnormality of the gallbladder wall was shown to be associated with reduced gallbladder contents/hepatic bile ratio of biliary lipids and of bilirubin. The concentrating function of the human gallbladder thus appears to be impaired in proportion to the severity of histologic lesions in its wall. Taken together with earlier findings in vitro, this relationship suggests impaired absorption of electrolytes and water by the gallbladder mucosa, or diffusion of biliary constituents from the lumen of the inflamed gallbladder.

Anticonvulsant-induced liver damage.

A case of iterated clonazepam-induced liver injury is described. It is suggested that the damage was of the metabolic idiosyncrasy type.

Cellular and serum immune response to dinitrophenol in adult Rana pipiens.

A rabbit anti-Rana anti-DNP antiserum was used in order to estimate changes in frequency of bone marrow (BM) cells containing cytoplasmic anti-DNP antibodies (cId+) for eight weeks following a single injection. We found that (i) cId+ cells increased from 0.2 to 4.3 percent of total mononuclear cells (MNC's) during the first 8 weeks, (ii) BM granulocytes increased in frequency up to week 2, then gradually decreased to week 8, and (iii) serum anti-DNP antibody levels, evaluated by ELISA, increased to week 8. Our observations (i) support those which suggest that Rana BM resembles mammaliam BM as a source of antibody-producing cells, (ii) indicate that specific immune responses in BM may result in a 20-fold increase in the frequency of antigen-related plasma cells among total MNC's, and (iii) suggest that BM B cell clonal expansion, maturation and secretion may result in an increase in specific antibody levels in serum.

Clinicopathologic studies on mesothelioma of the pleura. Differentiated approach to treatment.

A surgical series of 23 patients with pleural mesothelioma is reviewed. Three who had benign localized mesothelioma of fibrous type are alive and well at least 10 years postoperatively. In two others, radically extirpated localized mesothelioma was histologically classified as benign, but later proved to be malignant, causing death from recurrent disease 27 and 79 months postoperatively. Four patients with diffuse malignant mesothelioma underwent pleurectomy or open biopsy and survived for 2-9 months. Radical en-bloc pleuropneumonectomy was performed on 14 patients with diffuse malignant mesothelioma. One patient died postoperatively and the others succumbed to the disease after 3-51 (mean 20) months. The survival time was greater than or equal to 1 year in 62% of the patients and greater than 3 years in 23%. Patient age, histologic tumour type and extent of disease seemed to be important prognostic factors. Despite the generally poor prognosis, the results of radical surgery in this study appear to warrant an aggressive approach to treatment of benign or localized malignant pleural mesothelioma, and possibly also to stage I diffuse malignant mesothelioma of epithelial type.

Malignant mesothelioma in two pairs of siblings: is there a hereditary predisposing factor?

Two pairs of siblings with malignant pleural mesothelioma are reported. One sister and brother experienced slight household asbestos exposure during childhood. Two identical-twin brothers were occupationally exposed to asbestos for only 8 years. The occurrence of this rare neoplasm in 2 pairs of siblings indicates that a hereditary predisposing factor may exist.

Diagnosis and prognosis in malignant pleural mesothelioma: a prospective study.

In a prospective study of 336 consecutive patients with long-term pleural effusions, 32 cases of malignant mesothelioma were found. Microscopic examination of pleural tissue specimens, preferably selected at thoracoscopy, proved superior to pleural fluid analysis as an aid to correct diagnosis. The epithelial proved to be the most common type of malignant mesothelioma. As an example of the mesothelial cells' multipotent ability, malignant cells were seen transforming into fat-like cells with lipid-containing vacuoles. In the fibrous tumour type, cytological examination of pleural fluid revealed only normal cells. Cells with malignant features were seen in fluid samples from epithelial and biphasic tumour types. The malignant cells often formed tubuli-like aggregates which could be mistaken for adenocarcinoma. Hyaluronic acid was more frequently detected in tissue specimens than in the pleural fluid samples. The morphological type and the patient's age had an impact on the survival time, whereas sex and extensive surgical treatment seemed less important.