Rhodamine B fluorescence as a stain for amniotic fluid squames in maternal pulmonary embolism and fetal lungs.

Rhodamine B fluorescence is presented as a simple, rapid, highly sensitive, permanent method for the histologic detection of squames in maternal amniotic fluid emboli and fetal lungs in cases of intrauterine asphyxia. The method may be used on alcohol fixed smears or formalin fixed paraffin sections. The application of this procedure allows for identification of sparsely distributed fetal squames which otherwise may be overlooked by less sensitive tinctorial methods which may also be more tedious in technical preparation and screening.

Huge left atrial thrombus and valve degeneration in a patient with a bioprosthetic, porcine, mitral valve.

A 43 year old man with a Hancock porcine bioprosthetic valve in the mitral position developed a huge thrombus filling the entire left atrium and chronic degeneration of the bioprosthetic mitral valve. The effective valve orifice was less than 2 mm. These severe findings leading to the patient's death remained undetected while he was alive. This case illustrates the great need for a serial phonoechocardiographic studies in all patients with prosthetic valves.

Concentric membranous bodies in hepatocytes from a patient with systemic lupus erythematosus.

Concentric membranous bodies (CMB) are described in the cytoplasm of hepatocytes in a liver biopsy from a patient with systemic lupus erythematosus (SLE). Histologically, the biopsy showed binucleated hepatocytes and rosette formation with lymphocytic infiltrates in portal fields. Hepatocyte cytoplasm showed focal fatty metamorphosis, focal staining for RNA with azure B at pH 4.0 (which was perchloric acid labile), diffuse alloxan-Schiff staining for protein, and PAS-positive, diastase-sensitive glycogen. By electron microscopy one to three CMB were present in 10-15% of hepatocytes. CMB appeared as concentric, "fingerprint" or parallel arrays of particle-studded, membranous profiles. Other membranous configurations included mazelike forms. The space of Disse was dilated. Bile ductular changes included basement membrane thickening and redoubling and luminal bulging of ductular epithelium devoid of microvilli. CMB have been rarely reported in human hepatoma and have not heretofore been observed in non-neoplastic human liver. Their appearance in hepatocytes in a patient with SLE may reflect an increase in protein synthesis during regeneration.

Osteogenic sarcoma producing human chorionic gonadotrophin. Case report with immunohistochemical studies.

An osteogenic sarcoma occurring in a 22-year-old woman was found to be associated with production of a large amount of beta human chorionic gonadotrophin (B-hCG). Pregnancy was excluded on the basis of a normal ultrasonogram and a proliferative type endometrium obtained by curettage. A homogenate of the tumor was strongly positive for B-hCG while immunohistochemical staining of the tumor cells was strongly positive for B-hCG and negative for pregnancy associated glycoprotein. These results indicate ectopic production of hCG by the osteosarcoma.

Ultrastructural changes of intrahepatic bile ductules in griseofulvin fed mice.

Intrahepatic bile ductules in male Swiss Webster mice, fed a diet containing 2.5 percent griseofulvin (GF), showed marked hypertrophy and hyperplasia within two weeks. Thereafter, this reaction gradually increased in prominence until nine weeks. Cytoplasmic acid phosphatase and succinic dehydrogenase deposits were more prominent and abundant after 12 to 14 weeks. Ultrastructural changes in ductular epithelial cells included enlargement of nuclei and nucleoli and increased abundance of cytoplasmic rough surfaced endoplasmic reticulum (RER), ribosomes and mitochondria until nine to 12 weeks. At this interval, duct lumina appeared widely dilated with small clumps of electron dense material at the periphery adjoining apical cell membranes with severely flattened or absent microvilli. Electron dense needle-like crystals resembling protoporphyrin were clustered centrally in the lumina. After twelve weeks occasional mitochondria were markedly swollen and contained vacuolated areas. Numerous multivesicular bodies were noted. Mallory bodies were seen in a few duct cells. The spectrum of duct cell changes just noted gives further evidence of extensive liver injury after GF feeding. These changes, in addition to diverse hepatocyte alterations previously reported, constitute morphologic features of GF induced murine porphyria, also common to human porphyria and human alcoholic liver disease.

Focal nodular hyperplasia of the liver: ultrastructural observations.

Ultrastructural studies of the liver from a non-alcoholic 21 year old male with focal nodular hyperplasia (FNH) showed hepatocyte cytoplasmic alterations including swollen and bizarre mitochondria, rough endoplasmic reticulum (RER) in short parallel stacks or whorl-like arrangements, and Mallory bodies (MB). Proliferated bile duct cells contained occasional MB-like inclusions. Small hepatic blood vessels were dilated and some showed interruption and partial loss of their endothelial cell cytoplasm and thickened basement membranes. Liver tissue outside of the lesion appeared unaltered.

Ultrastructure and morphometric analysis of hypertrophic nucleoli in non-Hodgkin's lymphoma and phytohemagglutinin-stimulated lymphocytes.

The ultrastructure of hypertrophic nucleoli in large cells from each of 5 successive cases of non-Hodgkin's lymphoma (regardless of predominant cell type) showed irregularly rounded or ovoid, mostly compact forms with few scattered light spaces. Occasional nucleolonemal forms were noted. Few ring forms or transitional forms between ring and compact or nucleolonemal types were present. Granular components occupied 78.5 +/- 1.0% of the mean total area of 132 lymphoma large cell nucleoli of all configurations studied. Hypertrophic nucleoli in 27 phytohemagglutinin (PHA)-stimulated lymphocytes were predominantly nucleolonemal with 74.2 +/- 10.6% granular composition. The qualitative and quantitative distribution of nucleolar granular moieties did not vary significantly between individual cases of lymphoma nor quantitatively between these and nucleoli in PHA-stimulated lymphocytes. These observations lend further credence to the suggestion of the lymphocyte as a common progenitor cell type for the large B and T cell of malignant lymphomas. Some functional similarity of their nucleoli and those of PHA-stimulated lymphocytes in their participation in ribonucleic acid synthesis is implied. Generally compact nucleoli of lymphomas in contrast to nucleolonemal forms of PHA-stimulated lymphocytes may be pathognomonic for large lymphocytic cells undergoing neoplasia.

Identification of Mallory bodies with rhodamine B fluorescence and other stains for keratin.

Rhodamine B staining in conjunction with fluorescence microscopy is shown to demonstrate Mallory bodies. Mallory body morphology, localization, and distribution in hepatocytes from griseofulvin-fed mice, human hepatoma, and human alcoholics were similar to those observed in the same tissues after conventional staining methods for Mallory bodies. The presence of these inclusions was further confirmed by specific cytochemical localization with indirect immunoperoxidase labeling, horseradish peroxidase labeling, and electron microscopy. Other tinctorial or histochemical procedures previously used for keratin or prekeratin (modified Mallory stain, Kreyberg method, Pauly method for histidine) also stained Mallory bodies for study with white light microscopy but with decreasing sensitivity respectively. Mallory bodies from mouse and human liver both appear to contain a keratin-like moiety. This entity may be simply, rapidly, and permanently stained with rhodamine B, and selectively and reproducibly demonstrated with fluorescence microscopy.

RNA virus associated antigen in human placenta.

Using indirect immunofluorescence microscopy, it was demonstrated that human cytotrophoblast of six to ten weeks gestation contains mouse mammary tumor virus associated antigen. This antigen appeared predominantly in the periphery of the cells.

Hepatocyte mitochondrial alterations in griseofulvin fed mice.

Ultrastructural changes in mitochondria from hepatocytes of male Swiss Webster mice were observed during the course of porphyria induced by ingestion of a powdered diet containing 2.5 percent griseofulvin (GF). A variety of sporadic mitochondrial alterations were noted between three days and 22 weeks. Mitochondria appeared intact in hepatocytes from control mice fed powdered diet alone. The abundance and prominence of altered mitochondria increased after seven weeks and was particularly apparent after 15 weeks, concurrent with pronounced hepatomegaly and Mallory body (MB) formation. One or more changes observed in some mitochondria of several hepatocytes included: swelling, associated myelin bodies, increased matrix density, disoriented cristae, intracristal swelling, bizarre shapes, paracrystalline inclusions and elongated or spherulated giant forms. Many mitochondria showed intimate association with rough surface endoplasmic reticulum (RER) adjacent to smooth surface endoplasmic reticulum (SER). GF-induced mitochondrial defects appear to represent nonspecific morphologic alterations similar to many reported in human hepatocytes in alcoholic liver disease, porphyria and various other toxic and metabolic liver diseases. They may reflect another common site of metabolic insult between two such diverse processes as GF-induced mouse porphyria and human alcoholic liver disease.

Idiopathic intrahepatic cholestasis of pregnancy. Report of an unusual case and review of the recent literature.

An atypical case of idiopathic cholestasis of pregnancy occurring in the 16th week of gestation in a 24-year old Pakistani female is presented. The case is unusual both in the duration of the jaundice as well as in the high levels of bilirubin found in the presence of normal renal function. A review of the recent literature emphasizes that this disorder is not entirely benign for the mother and the fetus. It also discusses some of the alterations in estrogen and lipid metabolism that are seen, as well as the possible mechanisms responsible for the increased incidence of gallstone formation found in these patients.

Light and electron microscopy of hepatocellular changes in griseofulvin fed mice. Particular reference to Mallory bodies.

Male Swiss Webster mice were fed a diet containing 2.5 percent griseofulvin (GF). Serially progressive light and electron microscopic hepatic altertions were present from one day on and grossly visible hepatomegaly from two weeks until termination at 22 weeks. GF induced liver changes included hepatocyte nuclear and cytoplasmic hypertrophy, increased incidence of necrosis and mitosis, Kupffer cell activation, bile duct proliferation and portal fibrosis. Protoporphyrin crystals were present in hepatocyte cytoplasm as early as one day after GF feeding. Hepatocellular hyalin was initially noted at seven weeks. Thereafter, the hyalin increased in prominence and frequency of occurrence. Ultrastructurally, three types of hyalin have been presently demonstrated to correspond to Mallory bodies (MB) reported in human liver disease. Forms intermediate in appearance between various MB types suggested transition from one to another. Areas of organelle free cytosol with abundant, loosely scattered filamentous elements as well as vesicular, smooth surfaced endoplasmic reticulum and ribosomes appeared preceding and closely associated with MB formation. Similarities of hepatocellular MB observed in GF fed mice and reported earlier in human alcoholic liver disease suggest a common pathway in its formation as a response to divergent noxious insults.

Light microscopical examination of glomerular basement membrane in systemic lupus erythematosus.

To obtain as much information as possible from light-microscopy about changes in the glomerular basement-membrane, a thin section microtome attachment is necessary in order to produce 1 to 3 micron sections. The most informative staining method of paraffin embedded tissue is the Muller-Mowry-Masson trichrome (M-M-M) combination stain. The deposits, e.g., in systemic lupus erythematosus (SLE), stain red, the basement membrane light orange and the endothelial cell cytoplasm light pink. With this stain, as with the Muller-Mowry stain, normal glomeruli show an uninterrupted line of blue acid mucopolysaccharides along the basement membrane, where it is joined by foot processes of the epithelial cells. In contrast, in glomeruli from patients with SLE, focal loss of acid mucopolysaccharides is noted.

Percutaneous transhepatic cholangiography and needle biopsy in the differential diagnosis of obstruction of bile flow.

Twenty-four patients with clinical evidence of obstructive jaundice were examined by percutaneous transhepatic cholangiography (PTC) and needle biopsy (NB) of liver. The presence of extrahepatic bile duct obstruction was confirmed by surgery in 21 cases. PTC and the combination of both methods were superior to NB alone in the differential diagnosis between extra- and intrahepatic biliary obstruction. Sampling bias added to the difficulties of NB in distinguishing beween these two types of obstruction. The combination of both precedures proved most useful in three cases with intrahepatic obstruction, in which the patients were spared unnecessary surgery. In five cases the NB provided additional information about the nature of the tumor metastases and gave suport to the clinicians in their therapeutic approach.

Conjunctival biopsy in sarcoidosis: A simple, safe, and specific diagnostic procedure.

A prospective study was undertaken to determine the prevalence and specificity of granulomatous conjunctivitis in sarcoidosis. One hundred conjunctival biopsies were performed on patients with histologically confirmed sarcoidosis (60), pulmonary tuberculosis (25), and several nonspecific inflammatory diseases of the lung and eye (15). A positive biopsy was present in 33% of the patients with sarcoidosis, but in none of the other disease processes. Great care was taken to exclude the possibility of these granulomas being secondary to foreign body reaction or infection with tuberculosis or fungi. No predictive relationship could be established between eye symptoms or slit lamp findings in the conjunctiva and uveal tract, and the finding of positive conjunctival biopsies. Thus, it is suggested that this procedure, due to its ease, safety, and specificity, be the first biopsy performed in patients with clinical or chest x-ray abnormalities suggesting sarcoidosis.

Staining of minerals and solubility of iron in tissues.

Iron deposits in ethionine induced aortic siderosis of rats, in splenic deposits in sickle cell anemia and siderocalcific vessels in cerebral arteriosclerosis were completely removed by exposure to 20 percent hydrochloric acid for 30 min. This contrasted with idiopathic hemochromatosis and idiopathic pulmonary hemosiderosis in which the iron containing organs had to be exposed to 40 percent hydrochloric acid for two hours. The more soluble iron appeared colorless in unstained tissues, purplish blue with hematoxylin and eosin, turquoise blue with Perls' stain, violet blue with gallocyanin and dark-drown with sodium rhodizonate. The less soluble iron was golden yellow in unstained tissues. It appeared golden yellow with hematoxylin and eosin and sodium rhodizonate, but it stained greenish blue with Perls' method and dark brown with gallocyanin. Lead and copper were capable of deposition in some tissues in vitro in the presence of iron and/or calcium but not when these minerals were removed. This phenomenon may be of importance in certain pathological conditions, e.g. hemochromatosis, where on preexisting tissue-iron-complexes there is a secondary deposition of copper.