Luis Biempica

Collagen Types in Normal and Cirrhotic Liver

The purpose of this study was to characterize the collagen in normal liver and in liver cirrhosis of different etiology, and to determine whether the ratios of different collagen types vary with the agent that induced the disease. Collagens were extracted from five normal and eight cirrhotic livers, which were obtained at autopsy. Alcoholic, hemochromatotic, Wilsons disease, and posthepatitic cirrhosis were studied. The yield of extracted collagens was 60–95%. Normal liver contained 5.5 ± 1.6 mg of collagen/ g of wet tissue. In cirrhotic liver, collagen was increased four- to sevenfold. In all livers studied, approximately 2 mg of collagen/g of wet tissue remained insoluble. Each liver contained Type I and Type III collagen, and a group of collagens with amino acid composition similar to that found in basement membranes. Three of these basement membrane collagens, A, B, and E, representing onehalf of the liver basement membrane collagen, were purified and partially characterized. The other half, similar in composition to basement membrane collagen obtained from glomerulus, was heterogeneous and was not characterized. Collagens A and Bare identical to the A and B chains isolated from placenta and skin. Collagen E is similar to the collagen previously isolated from human aorta. In cirrhotic, but not normal liver, an additional collagen, which was identified as a trimer of α1 Type I collagen, was found. Normal livers contained approximately equal amounts of Type I, Type Ill, and basement membrane collagens; The ratio of Type l/Type III collagen was smaller than 1, suggesting a slight predominance of Type III collagen. In cirrhotic livers regardless of etiology, all collagen Types were increased; in livers containing 20 mg of collagen/g of tissue, Type I collagen was the predominant type, and the ratio of Type I/Type III collagen was increased.

ULTRASTRUCTURAL AND CYTOCHEMICAL OBSERVATIONS ON B-16 AND HARDING-PASSEY MOUSE MELANOMAS THE ORIGIN OF PREMELANOSOMES AND COMPOUND MELANOSOMES

The B-16 and Harding-Passey mouse melanomas were studied by light microscopy (tyrosinase, acid phosphatase, aryl sulfatase, thiamine pyrophosphatase and inosine diphosphatase activities) and electron microscopy (morphology and tyrosinase and acid phosphatase activities). Lysosomal enzyme activity is present in individual premelanosomes and melanosomes as well as in compound melanosomes. Acid phosphatase and tyrosinase activities are present in a Golgi-associated system of smooth endoplasmic reticulum (GERL) and small vesicles related to it. The acid phosphatase and tyrosinase activities of premelanosomes, and morphologic appearances, support the hypothesis that the granules arise from GERL. On the basis of the evidence presented, it is suggested that compound melanosomes arise within melanoma cells by autophagy.

Clinical utility of liver biopsy in patients with serum antibodies to the human immunodeficiency virus

PURPOSE Patients with the acquired immunodeficiency syndrome (AIDS) frequently have liver dysfunction, which may be due to a number of causes. Determination of the patients who are likely to benefit from liver biopsy, an invasive procedure, is therefore important. In this study, the results of liver biopsy in patients with AIDS were compared to those in human immunodeficiency virus (HIV)-infected patients without AIDS. PATIENTS AND METHODS Thirty-six consecutive patients with antibodies to HIV present in the serum underwent liver biopsy from 1984 through 1988 at the Bronx Municipal Hospital. Twenty (56%) of the patients had AIDS diagnosed prior to the liver biopsy. Indications for the liver biopsy were unexplained fever in 83%, and abnormal serum levels of biochemical parameters of liver function in 89%. RESULTS Liver biopsy was diagnostic in 18 cases (50%), including findings of hepatic infection by mycobacteria in 15, cytomegalovirus in two, and schistosoma in one; these infections had been previously detected at an extrahepatic site in only two cases. Helpful clinical information in 10 others included findings of granulomas of undetermined etiology in four, cirrhosis in five, and chronic persistent hepatitis in one. Patients with a diagnostic biopsy, as compared to patients with a nondiagnostic biopsy, had a statistically significant increase in the frequency of having AIDS diagnosed before the biopsy, longer duration of AIDS (in patients with AIDS diagnosed before the biopsy), greater number of different prior opportunistic infections, and a more elevated serum alkaline phosphatase level. For example, 70% of patients with AIDS, as compared to 25% of patients with serum antibodies to HIV but without AIDS, had diagnostic liver biopsies. Patients with a diagnostic biopsy also had statistically significantly more frequent pulmonary symptoms, possibly due to more frequent occurrence of Pneumocystis carinii pneumonia. In particular, the 15 patients with hepatic mycobacterial infection, as compared to the other patients, had a statistically significant increase in the frequency of having AIDS diagnosed prior to the biopsy, longer duration of AIDS, more frequent prior opportunistic infections, more severe leukopenia, and a more elevated serum alkaline phosphatase level. Liver biopsy was more sensitive than bone marrow aspiration and biopsy at detecting mycobacterial infection. CONCLUSION Liver biopsy, when indicated, is useful to detect opportunistic infection in HIV-infected patients who are not known to have AIDS.

Carcinoma of the breast with multinucleated reactive stromal giant cells

Two unusual carcinomas of the breast are described, containing nests of infiltrating neoplasm situated within stromal lacunar spaces, and surrounded by numerous benign appearing multinucleated giant cells. Within the stroma, there was extensive hemorrhage, hemosiderin pigment deposition, and large numbers of mononucleated inflammatory cells. The morphology of both tumors resembled the giant cell tumor of bone. Although a similar giant cell reaction has recently been described in association with a uterine leiomyosarcoma, we are aware of only two other examples of this entity in the breast, both reported over 40 years ago in the French literature. This is the first report in which electron microscopy confirmed the benign histiocytic nature of the giant cells. These cells had many of the ultrastructural features of multinucleated giant cells described in tissue culture, skeletal osteoclastomas, and foreign body granulomas. We propose that the giant cells arise from fusion of mononucleated stromal cells, and most likely are reactive histiocytic elements which are in some way related to the tumor cell nests. Further studies of these unusual neoplasms are needed to determine if the giant cell reaction in any way affects the prognosis of the patient.

The Effects of Hepatic Fibrosis on Ito Cell Gene Expression

While Ito cells appear to be a major source of increased matrix synthesis during hepatic fibrogenesis, the cellular changes that occur in these cells during liver fibrosis have not been well delineated. In this study we examined Ito cell gene expression in isolated cells from normal rats, and rats with carbon tetrachloride-induced fibrosis, in order to better define the changes occurring in these cells during this pathologic process. Specifically, we addressed three questions: (1) which matrix genes are over expressed in Ito cells in fibrotic liver; (2) do these cells increase their expression of the fibrogenic cytokine transforming growth factor-beta 1 (TGF-beta 1); and (3) do Ito cells change their phenotype during hepatic fibrogenesis as reflected by alterations in the expression of their intermediate filament genes? Northern hybridization analysis revealed that Ito cells isolated from fibrotic livers had significant increases in mRNA levels of types I, III and IV procollagen compared to normal cells, while no increases were found in hepatocytes, and Kupffer/endothelial cells had only an increase in type I procollagen mRNA. Analysis of other matrix proteins which increase during hepatic fibrogenesis revealed elevations in laminin B and fibronectin mRNA levels only in Ito cells. Increased Ito cell matrix gene expression was also associated with a 4-fold increase in TGF-beta 1 levels in these cells. No increase in TGF-beta 1 mRNA was found in hepatocytes, and less than a 2-fold increase was found in Kupffer/endothelial cells isolated from fibrotic livers.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphological and Biochemical Studies of Benign Recurrent Cholestasis

Summary A 27-year-old Caucasian man had experienced 14 episodes of pruritus, jaundice, bilirubinuria, and weight loss since puberty. Remission occurred spontaneously on each occasion. During a clinical exacerbation, bilirubin, alkaline phosphatase, and serum di- and trihydroxy bile acid concentrations were increased and Tm sulfobromophthalein and hepatic sulfobromophthalein storage were reduced. Examination of a surgical biopsy specimen taken at that time revealed markedly altered bile canaliculi, with distorted and reduced microvilli, almost complete disappearance of nucleoside phosphatase activity, and reduction in the number of acid phosphatase-rich lysosomes. Lipid spheres and glycogen nuclei were abundant and intracanalicular, and intracellular bile pigment was common. During clinical remission, 10 months later, functional, cytochemical, and ultrastructural studies showed a remarkable return to normal patterns. Three exceptions were noted: the presence of small amounts of intracellular and intracanalicular bile pigment, some glycogen-rich nuclei, and the presence of crystalloid nucleoids within the microbodies.

Coronary intimal sclerosis in Morquio's syndrome.

Mitral valve, coronary arteries, cartilage, and liver were studied by light and electron microscopy in a 15 year old boy with Morquios syndrome, a genetic mucopolysaccharidosis, in which a deficiency of lysosomal hexosamine sulfatase is associated with accumulations of keratan sulfate in various organs. Coronary artery intimal sclerosis was a prominent feature of this disorder. Ultrastructural examination revealed numerous intimal smooth muscle cells containing storage vacuoles consistent with lysosomes. This was associated with marked interstitial deposition of collagen, elastin, and basement membrane material. Recent studies of human and experimental atherosclerosis have demonstrated the accumulation of cholesterol within vascular smooth muscle cell lysosomes. Intralysosomal accumulation of substrates other than cholesterol is also associated with vascular intimal sclerosis in genetic lysosomal disorders such as Fabrys disease and Hurlers syndrome. Lysosomal storage of undegraded substrate may be an important pathogenetic mechanism in the development of sclerotic vascular lesions.

Immunohistochemical localization of collagenase in hepatic murine schistosomiasis.

It has been previously demonstrated that collagenase activity and collagen synthesis in hepatic granulomas of mice infected with S. mansoni cercariae are maximal 8 weeks after infection; however, total liver collagen content continues to increase. Now the anatomic relationships among collagenase and collagen, granulomas, and hepatic parenchyma in normal mice and in mice infected with S. mansoni are studied. Trypsin-activated collagenase was purified from the media of cultured granuloma explants and anti-collagenase immunoglobulin G was purified from immunized rabbits. The IgG cross-reacted with liver granulomas and active and inactive forms of collagenase, but did not react by immunodiffusion in agar with other neutral proteases or homogenates of schistosome eggs or normal liver. Cryostat sections of liver from normal and infected mice were studied by indirect immunohistochemical methods using fluorescein, rhodamine, and peroxidase labels. Collagenase localization was restricted to areas of collagen deposits in granulomas and hepatic parenchyma. Ultrastructural studies revealed collagenase on the surface of collagen fibers. Hepatocytes of normal mice showed delicate staining at the sinusoidal surface. At all times, immunoreactive collagenase was intimately associated with its substrate, where it presumably initiated collagen degradation. This localization provides a rationale for possible therapeutic approaches to control fibrogenesis through collagenase induction or activation.

Primary cardiac neurilemoma

A rare case of primary cardiac neurilemoma arising in the right atrium is reported. The patient was a 55‐year‐old woman who was treated for 20 years with drug and radiation therapy for a metastatic ovarian carcinoma. The cardiac tumor was an incidental finding at autopsy. Both light and electron microscopic examination of the tumor confirmed the Schwann cell origin of the neoplasm. Review of the literature reveals only one other unequivocal report of a primary cardiac neurilemoma.

Esophageal perforation at a Barrett's ulcer

An alcoholic man with known reflux esophagitis and Barretts esophagus developed fever, epigastric pain, subcutaneous crepitus, and leukocytosis from an esophageal perforation at a Barretts ulcer. Possible risk factors for perforation in this patient included alcoholism, severe gastroesophageal reflux, corticosteroid therapy, noncompliance with antacid and H2 blocker therapy, and the presence of acid-secreting parietal cells in the Barretts epithelium. Five cases of this complication have previously been reported in a review of the literature, which included 536 cases of Barretts esophagus or esophageal perforation. This entity may present with a clinical triad of a patient (a) in acute distress with fever and epigastric or noncardiac chest pain and without signs of peritonitis, (b) with symptoms of or known gastroesophageal reflux, and (c) with chest examination revealing subcutaneous crepitus, or chest roentgenogram revealing subcutaneous emphysema, pneumomediastinum, or hydropneumothorax.