Primary ovary choriocarcinoma: individual DNA polymorphic analysis as a strategy to confirm diagnosis and treatment.

Primary choriocarcinoma of the ovary is rare. Furthermore, this tumor can arise from gestational tissue or pure germ cells of the ovary, with the latter resulting in non-gestational choriocarcinoma. While the clinical characteristics and histology of both tumor types are identical, differentiation of these tumors is necessary for effective treatment. One strategy for the differentiation of these tumors types is to assay for the presence of paternal DNA. Accordingly, in the present case, a patient with primary choriocarcinoma of the ovary with a non-gestational origin was confirmed by DNA analysis. The patient subsequently exhibited an excellent response to chemotherapy, and following surgery, achieved complete remission. A pathological analysis of surgical specimens further confirmed the absence of tumor.

Urinary biomarkers in charcoal workers exposed to wood smoke in Bahia State, Brazil.

Charcoal is an important source of energy for domestic and industrial use in many countries. Brazil is the largest producer of charcoal in the world, with approximately 350,000 workers linked to the production and transportation of charcoal. To evaluate the occupational exposure to wood smoke and potential genotoxic effects on workers in charcoal production, we studied urinary mutagenicity in Salmonella YG1041 +S9 and urinary levels of 2-naphthol and 1-pyrenol in 154 workers of northeastern Bahia. Workers were classified into three categories according to their working location, and information about socio-demographic data, diet, alcohol consumption, and smoking was obtained using a standard questionnaire. Spot urine samples were collected to evaluate urinary mutagenicity and urinary metabolites. Urinary mutagenicity increased significantly with exposure to wood smoke and was modified by smoking. The prevalence odds ratio was 5.31, and the 95% confidence interval was 1.85; 15.27 for urinary mutagenicity in the highly exposed group relative to the nonexposed group. The levels of urinary metabolites increased monotonically with wood smoke exposure and were associated with the GSTM1 null genotype, which was determined previously. The prevalence odds ratio (95% confidence interval) for higher levels of 2-naphtol among the highly exposed was 17.13 (6.91; 42.44) and for 1-hydroxyprene 11.55 (5.32; 25.08) when compared with nonexposed workers. Urinary 2-naphthol was the most sensitive indicator of wood smoke exposure. This is the first reported measurement of internal exposure to wood smoke among charcoal workers, and the results showed that these workers receive a systemic exposure to genotoxic compounds.

Extravascular haemolytic anaemia in chicks infected with highly pathogenic Marek's disease viruses.

Chicks were infected at 1 day of age with highly pathogenic AC-1 or RB-1B isolates of Marek's disease virus and 14 to 36 days post-exposure they were bled for haematocrits and euthanized. At necropsy, specimens were collected for light and electron microscopy. Both viruses caused significant reductions in packed cell volume (P<0.01). Hyperplasia of reticuloendothelial (RE) cells in the liver was a common finding in chicks that were ill from infection with AC-1 or RB-1B viruses. Erythrophagocytosis by sinusoidal macrophages was common and some of these cells contained an overload of haemosiderin. RE cells in the spleen were also active in destruction of erythrocytes. In chicks that were severely affected by AC-1 virus(s) there were often groups of RE cells in liver that had undergone lytic necrosis. Some of these foci appeared to have been replaced by proliferative lesions that ranged from being lymphogranulomatous to neoplastic. Tumour development was more apparent with RB-1B than with AC-1 and the multiple tiny lymphomas observed in RB-1B infected livers appeared to have developed from sinusoidal mononuclear cells. Ultrastructurally, Kupffer cells often contained stacks of electron light cisternal tubules that were densely packed with parallel fibrils. There were also groups of short electron opaque cisternal tubules that sometimes contained ghosts of fibrils packed in parallel. These organelles gave positive reactions for catalase and peroxidase and might have been peroxisomes that developed from the cisternal system due to the overload with iron. It was concluded that extravascular haemolytic anaemia resulted from erythrophagocytosis by hyperplastic RE cells.

Cytopathology caused by the AC-1 isolate of Marek's disease virus in the feather follicle epidermis.

The AC-1 strain of Marek's disease herpesvirus (MDV), recently isolated from an outbreak of Marek's disease in vaccinated chickens, was used for intraperitoneal inoculation of one-day-old Single Comb White Leghorns. Chicks were necropsied 12 to 32 days post inoculation and skin was collected for electron microscopic studies. This paper focuses on early stages of MDHV development in feather follicle epithelial cells and on cytoplasmic changes that occur in formation and release of virus particles. In some nuclei we observed helical threads of DNA which were partially encircled by capsid membranes. This was interpreted to be an early stage of nucleocapsid development. In the infected epidermal cells, the smooth endoplasmic reticulum vesiculated and appeared to be hyperplastic. Immature virus particles released into cytoplasm acquired their outer envelope while budding into cytoplasmic vesicles. Vesicles containing mature viruses were referred to as cytoplasmic vesicular inclusions. It appeared that the degenerative process in cells facilitated the release of virions from the feather follicle epithelium.

Response of meat-type chickens to infection with RAV-1 avian leukosis virus.

Meat-type and White Leghorn chickens were inoculated with the RAV-1 strain of avian leukosis virus at 1 day of age and the severity of infection was assessed by clinical illness, haematology and post-mortem findings. The following were examined from selected birds: histological section for chronic mononuclear myocarditis, immunohistochemically-stained sections of myocardium, spleen, bursa of Fabricius and kidney for group-specific viral antigen, and ultrathin sections of these tissues for virus particles by electron microscopy. The experiment was terminated at 115-122 days. Approximately one-third of the 52 inoculated White Leghorns appeared anaemic at 3-4 weeks of age whereas there was no evidence of anaemia in 177 inoculated meat-type birds or in uninoculated birds of either type. Haemograms confirmed these observations. The first tumour found was a myelocytoma and it was in a meat-type bird at 65 days. Of the 151 meat-type birds of the inoculated group alive at 65 days, 8.5% developed nephroblastomas, but there were no cases of lymphoid leukosis. Myocarditis and virus replication in myocardium were usually more extensive in chickens that developed nephroblastomas than in those without such lesions. In 17 uninoculated control chickens examined between 26 and 122 days of age there were no virus particles or lesions in myocardium.

Chronic myocarditis and circulatory syndrome in a White Leghorn strain induced by an avian leukosis virus: light and electron microscopic study.

Specific-pathogen-free white leghorn chickens were inoculated at 1 day of age with avian leukosis virus (ALV, RAV-1). All chickens in Expt. 1, killed 33 or 64 days postinoculation, had focal chronic lymphocytic or lymphoplasmacytic myocarditis. Among those held beyond 33 days, eight of 22 developed lesions in the myocardium that resulted in a chronic circulatory syndrome (CCS) typical of right-sided heart failure. Chickens in Expt. 2 were held for 210 days, and 21% of 125 developed CCS. In Expt. 2, ALV particles were found by electron microscopy in myocardium of 100%, 72%, and 89% of inoculated chickens that developed CCS, lymphoid leukosis, or that had no gross lesions, respectively. These findings were in accord with the immunoperoxidase staining of tissue sections for group-specific antigen of ALV. In areas of extensive virus replication, there were often abnormal virus particles and also round bodies, which may have been remnants of host-cell membranes formed in the budding process. In contrast to findings in hearts, the spleens were usually negative for virus and viral antigen.

Importance of the medullary macrophage in the replication of lymphoid leukosis virus in the bursa of Fabricius of chickens.

Electron microscopy and immunocytochemistry were used to study the development of lymphoid leukosis virus infection in the bursa of Fabricius of experimentally infected chicken embryos and chickens. In embryos infected at 7 days of incubation and killed 10 days later, virus particles and group-specific viral antigen were confined mainly to the connective tissue of the lamina propria of the bursal mucosal folds; a few developing follicles had discrete virions and group-specific antigen between cells. In chickens infected at 1 day of age, infection (as determined by use of electron microscopy and immunocytochemistry) was maximal in 1- to 4-month-old birds, and the greatest concentration of virus and group-specific viral antigen was in the medulla of the follicles. Although lymphoid leukosis virus was released from lymphocytes, epithelial cells, and macrophages, virus replication in the medullary macrophages was more active than that in the other cells. Normal medullary macrophages had cell membrane vesicles (50 to 80 nm in diameter) that covered part of all of the cell membrane surface. In infected chickens, virus particles frequently developed within these vesicles. Comparable vesicles were not found on cortical macrophages. Results of the present study indicated that the medullary macrophage was the principal host cell for replication of lymphoid leukosis virus in the bursa of Fabricius of the chicken.

Feather pulp organ cultures for assessing host resistance to infection with avian leukosis-sarcoma viruses.

The purpose of this study was to improve in vitro procedures for detecting cellular resistance to the avian leukosis-sarcoma group of viruses. Four feather pulp organ cultures (FPOC) were prepared from each chicken by placing pulp squeezed from feathers in wells of microtitre plates that contained culture medium. Two of the four FPOC were inoculated with Rous sarcoma virus (RSV) of subgroup A and 5 to 6 days later the fluids from all four cultures were assayed for virus by inoculating chicken embryo fibroblasts (CEF) and examining for development of foci of transformed cells. Prior to the second assay of culture fluids, quail cells transformed by envelope-defective RSV [R(-)Q cells] were added to some RSV-inoculated and uninoculated FPOC. The R(-)Q cells produce infectious RSV when infected with avian leukosis virus (ALV), and hence made it possible to detect ALV in FPOC. Status of host infection was also assessed by tests for virus neutralising antibody and the enzyme-linked immunosorbent assay for group specific viral antigen. In one experiment FPOC from chickens not exposed to ALV were susceptible to RSV throughout the 140-day test period. In contrast, FPOC from ALV-inoculated chickens were usually infected with ALV and were resistant to RSV. FPOC from chickens reared in contact with the inoculated group for 121 days were free of ALV and were unexpectedly resistant to RSV. Two other experiments supported the observation that genetically susceptible chickens acquire cellular resistance to RSV as a result of persistent or transient ALV-infection. In Cornell K strain chickens there was close agreement between cellular susceptibility based on tests on FPOC prepared prior to inoculation of chickens with ALV and for antibody following inoculation with ALV. A New Hampshire strain showed a high degree of genetic cellular resistance by these test procedures.

Viral matrix inclusion bodies in myocardium of lymphoid leukosis virus-infected chickens.

Chicken embryos and healthy adult chickens naturally infected with lymphoid leukosis virus were used to investigate viral inclusion bodies in myocardial cells by light and electron microscopies and by immunocytochemical technique. Intracytoplasmic viral matrix inclusion bodies frequently appeared in the myocardium of adult chickens, but not in that of embryos. In light microscopic preparations, inclusions were irregularly distributed, were basophilic, and contained ribonucleic acid. Ultrastructurally, inclusions in myocardial cells were in areas containing numerous interstitial C-type particles. Early inclusions were composed of clusters of ribosomes associated with sarcoplasmic tubules; spherical bodies developed among these ribosomes. Mature inclusions were composed of numerous spherical bodies (50 to 75 nm) with interspersed ribosomes and of ribosomes clustered at the periphery. Inclusions were not membrane-enclosed. Occasionally, spherical bodies were in paracrystalline arrays. Multiple budding occurred on cell membranes adjacent to matrix inclusions. The viral group-specific protein, p27, was demonstrated by the peroxidase-antiperoxidase method and by the protein A-gold method in the spherical bodies, in nucleoids of mature virus particles, and among ribosomes of inclusions. The results indicate that the matrix inclusions were the result of lymphoid leukosis virus infection and were the product of viral protein synthesis on ribosomes.

Immunohistochemical identification of group specific antigen in avian leukosis virus infected chickens.

An indirect immunoperoxidase method was employed in the detection of the group specific antigen of avian leukosis virus in the oviduct, spleen, myocardium and bursae of Fabricius of chickens. In the magnum of the oviduct the group specific antigen was detected at the base and in the lumina of glands. In the spleen the group specific antigen was found in and around the arterioles, sheathed capillaries, and in the capsular tissue. In the myocardium the group specific antigen was present in the intercellular spaces and also in some myocardial cells. The bursa of Fabricius had concentrations of the group specific antigen in the medullary parts of follicles over the surface of large lymphoid cells. Absolute ethyl alcohol with acetone and modified Bouin's fluid were suitable as fixatives for preserving the group specific antigen in specimens embedded into low melting point paraffin.

Distribution of lymphoid leukosis virus and p27 group-specific antigen in tissues from laying hens.

In order to gain insight into transmission and pathogenesis of infection, specimens from laying hens that had been naturally exposed to lymphoid leukosis virus (LLV) were tested for group-specific antigen (gsa) of the virus by immunofluorescence (IF), complement fixation (CF), and the enzyme-linked immunosorbant assay (ELISA). Electron microscopic examinations determined the distribution of C-type virus particles in tissues, and the phenotypic-mixing test served as a biological assay for exogenous LLV. The IF gsa was found in all organs tested, and fluorescence was usually found where virus particles were concentrated. In the oviduct and intestine, IF gsa was frequently at the border of the lumina and in the connective tissue associated with basal membranes of glands. In skin, the antigen was detected in smooth muscle, in feather pulp, and in basal epidermal cells of developing feathers. Results of various tests on Ottawa strains of chickens were usually in agreement. For example, among hens that shed gsa into egg albumen, only the viremic hens were consistently positive for IF gsa in both spleens and oviducts. Geometric mean CF titers of antigen were respectively five- and 23-fold higher in spleens and oviducts from viremic hens than in those from nonviremic hens. These findings suggest that the gsa was associated with exogenous virus infection. In Cornell S strain hens that had not been exposed to LLV, gsa was detected in splenic tissue by CF and ELISA but not by the IF test. This gsa was presumed to be of endogenous origin.

Ectopic mineralization and nutritional hyperparathyroidism in boars.

The influence of a high phosphorus (1.5%) and high calcium (2.2%) diet on ectopic mineralization in boars was examined over a four month period. The high phosphorus diet caused metastatic mineralization in the left atrial endocardium in 84% of animals and in the pulmonary and diaphragmatic pleura in 21 and 58% of animals, respectively. Mineralization, that apparently commenced as deposits of extraneous calcium and progressed by metastatic deposition, occurred also in the lamina propria mucosae of the respiratory airways and in the lamina muscularis mucosae of fundic stomach. Hyperplasia of osteoclasts and microfractures in costochondral junction, morphological features of activated parathyroid cells and a significant drop in serum phosphorus during the fourth month in boars fed high phosphorus, suggest that nutritional hyperparathyroidism was experimentally induced. No systemic bone disease developed. Feeding a high calcium diet resulted in 20% incidence of discrete lesions in the left atrial endocardium and no pleural involvement. Also, lesions in respiratory airways and fundic stomach were mild. The fact that high phosphorus but not high calcium increased the incidence and extent of ectopic mineralization suggests that hyperparathyroidism under normocalcemic conditions might be involved in the pathogenesis of these lesions.

Electron microscopy of radiating clubs in tonsillar crypts of swine.

Tonsils of 25 adult swine with inflamed crypts were screened for the presence of radiating clubs, and five were selected for electron microscopic study of pathogenesis of clubs. Radiating clubs in crypts were surrounded by an exudate which principally contained neutrophils. Clubs were present on the outer surface of bacterial microcolonies; they also were present on the outer surface of plant particles wedged in crypts or on the cell walls of these particles near bacterial microcolonies. The study demonstrated development of clubs from an amorphous material derived from bacterial and leukocytic cells and also from precipitated proteins. The amorphous material was shaped into clubs by a protracted phagocytic activity of numerous neutrophilic leukocytes. The lesions with radiating clubs within crypts were not surrounded by granulomatous inflammation due to the intact epithelial lining of crypts, and therefore they differ from the granulomas having clubs around microbial colonies. They could, however, serve as models of pathogenesis of radiating clubs in the purulent core of specific granulomas with radiating clubs.

Detection of lymphoid leukosis virus infected chickens by testing for group-specific antigen or virus in feather pulp.

Exogenous lymphoid leukosis virus (LLV) and group-specific antigen (gsa) were detected in feather pulp and other specimens from naturally or experimentally infected chickens by phenotypic mixing or complement fixation tests, respectively. Electron microscopic studies on the calamus of plucked feathers revealed numerous C-type virus particles in intercellular spaces of the epidermis and pulp. LLV and gsa were detected in feather pulp from approximately 90% of 47 newly-hatched chicks that were shown to be congenitally infected. Chicks that were inoculated at 1-day-old or reared as contact controls were positive for gsa in feather pulp at 44 days, whereas at 111 days only the inoculated chicks were positive. Isolated controls were consistently negative. Titres of gsa /= 1 : 64. It was concluded that gsa could be detected in the feather pulp of most chickens actively infected with LLV and that titres would generally be higher in exogenous than in endogenous infections.

Lymphoid leukosis: detection of group specific viral antigen in chicken spleens by immunofluorescence and complement fixation.

Monospecific antiserum obtained from rabbits hyperimmunized against homogeneous p27 group specific protein purified from avian myeloblastosis virus was commercially procured and was then conjugated with fluorescein isothiocyanate. The conjugate was applied to spleens from naturally or experimentally infected chickens that had no evidence of lymphoid tumors. Fluorescence was usually localized in connective tissue of sheathed capillaries giving it a ring-like appearance. Sites of fluorescence corresponded to sites of greatest virus concentration as detected by electron microscopy, indicating that in such cases the group specific antigen may have been associated with virus particles. The group specific antigen could also be detected in the spleen by complement fixation and results of this test usually agreed with the immunofluorescent test and with the phenotypic mixing test which detects exogenous lymphoid leukosis virus.

A mouse model for estimation of Pasteurella haemolytica deposition in calf lungs following aerosol exposure.

A method used to calculate the number of Pasteurella haemolytica reaching the lungs of calves during an aerosol exposure is described. This method is based on a linear relationship of bacterial deposition in lungs of mice and calves when exposed to the same bacterial aerosol.

Focal mineralization and nonspecific granulomatous inflammation of respiratory mucous membranes in pigs.

Discrete mineralized foci and granulomatous inflammation occurred in the lamina propria mucosae of respiratory mucous membranes of adult pigs. Lesions were present in clinically healthy pigs of both sexes, including castrated males, fed various pelleted or non-pelleted diets. They were mainly in longitudinally corrugated mucosae of the dorsal wall of the trachea. Identical mineralization and inflammation occurred in the nasal cavity and, in decreasing frequency and intensity, in the thoracic trachea and bronchi. The lesions in respiratory mucous membranes occurred in pigs with and without mineralization in other organs. The distribution of lesions in the respiratory tract, and the higher frequency in pigs fed non-pelleted dusty feeds, suggest that focal mineralization was caused by inhaled particles of calcium salts.

Experimental ectopic mineralization in young boars I. The role of supplemental vitamin D inthe etiology.

Young boars supplemented orally with 800, 500,300 and 100 IU of vitamin D2/kg of feed and control boars not supplemented with vitamin D2 for four months developed mineralization in the left atrial endocardium, lamina muscularis mucosae of the fundic stomach and other sites. Since low levels of supplementation with vitamin D2 did not eliminate the lesions, the levels of vitamin D2 added appeared not to be involved in the pathogenesis. All boars had mild hypercalcemia throughout the experiment, and phosphorus levels in sera were lower in all animals receiving than in those not receiving calciferol. Mild morphological lesions of rickets developed in several boars not receiving supplemental vitamin D2 and in boars supplemented with 100 an 300 IU/kg of feed.

Heterotopic calcification in swine.

Pigs on nutritional studies developed heterotopic calcifications in the left atrial endocardium, submucosa of the gastric fundus and to a lesser extent in other sites. Light and electron microscopy of mineralized tissue showed early edema and connective tissue change, deposition of calcium, granulomatous reaction and repair. There was no alkaline phosphatase activity at the sites of calcification and the distribution of this enzyme activity was normal in other tissues. Increased acid phosphatase activity was demonstrated only in the lysosomes of macrophages in areas of granulomatous inflammation. Calcium and magnesium were within normal range in serum and femoral bone. The character and distribution of the lesions suggested the possibility of protracted low grade hypervitaminosis D3.

Pathology in rabbits treated with leukocyte-degraded meningococci in combination with meningococcal endotoxin.

The effects of a preparative dose of the leukocyte egesta containing degraded meningococci and a provocative dose of the meningococcal lipopolysaccharide on development of pathological lesions associated with disseminated intravascular coagulation were studied in tissues of 32 rabbits. These effects were compared with effects of a single dose of meningococcal lipopolysaccharide as well as leukocyte egesta containing degraded Staphylococcus epidermidis. Rabbits injected subcutaneously with egesta containing degraded meningococci followed after 12 h with meningococcal endotoxin (intravenously) exhibited heterophilic leukocytosis and disseminated intravascular coagulation mainly in the pulmonary capillaries and venules; focal necroses occurred in myocardium, lungs, and liver, whereas, cortical renal necrosis developed in lethal cases. Similar lesions, however, but less severe and with less frequency, developed even after a single dose of meningococcal endotoxin or after endotoxin that followed a dose of supernatant fluid from normal leukocytes. Our findings suggest that meningococcal material from polymorphonuclear degradation plays a role in the pathology characteristic of meningococcal septicemia.