PubMed Health⌕ Search

Biomedical subjects

J Alroy

Publications and source records attributed to J Alroy.

At least 55 records · Page 3Linked to original sources

Eosinophil granulocytic sarcoma in a New Zealand white rabbit.

An 18-month-old New Zealand White rabbit developed a rapidly growing firm perianal mass. Within 2 months, the mass increased in size from 4.0 x 2.0 cm to 7.5 x 7.0 x 3.5 cm. Cut surfaces of the mass were white to yellow with foci of hemorrhage. Histopathologic examination revealed neoplastic granulocytes that invaded the dermis of the overlying skin, subcutaneous tissue, and the regional skeletal muscle. The tumor was comprised of myeloid cells with large eosinophilic cytoplasmic granules that had the histochemical and ultrastructural features of eosinophils. This is the first report of an eosinophil granulocytic sarcoma in lagomorphs.

Animals↗

Electron microscopic examination of skin biopsy as a cost-effective tool in the diagnosis of lysosomal storage diseases.

In this report, we have summarized our 9-year experience of over 100 proven cases of lysosomal storage disease using electron microscopic evaluation of skin biopsies as a screening tool. The skin biopsy was very specific in establishing the diagnosis in only two disorders, namely neuronal ceroid lipofuscinosis and mucolipidosis IV. Although the biopsy was not diagnostic in other categories of storage diseases, it proved to be highly sensitive and provided valuable clues to direct further investigations on the basis of morphologic appearance of stored material and the cell type affected. Only in two cases of biochemically proven lysosomal storage disease was the morphologic diagnosis unable to be confirmed. We have compared the cost of screening for storage disorders using skin biopsy with the cost of performing multiple lysosomal enzyme assays. Our findings indicate that the skin biopsy, although more expensive than a single enzyme assay, provides an efficient, rapid, cost-effective tool to screen for more than 35 lysosomal storage disorders.

Biopsy↗

Cutaneous amyloidosis in a horse with lymphoma.

A horse with malignant lymphoma (histiolymphocytic) and cutaneous amyloidosis is described. The lymphoma involved the dura mater of the spinal cord and some of the peripheral lymph nodes. Multifocal amyloid deposits were present in the skin and subcutis of the ventral abdomen but not within the lymphoma cell infiltrates or in the viscera.

Amyloidosis↗

An in vitro model for abnormal skeletal development in the lysosomal storage diseases.

Lysosomal storage diseases such as GM1-gangliosidosis are associated with skeletal abnormalities. Radiological and histological studies, both in human and corresponding animal models, indicate retarded bone formation. Since cartilage maturation leads to bone formation, we developed an in vitro system to study and compare the biological features of cartilage from dogs affected with GM1-gangliosidosis with age-matched controls. Costochondral chondrocytes were grown in monolayer and in agarose culture. Both affected and control cells dedifferentiated in monolayer; however, in agarose culture they re-expressed the chondrocytic phenotype. Cells from affected dogs were enlarged and contained numerous large vacuoles when compared with control cells. This morphology was similar to that seen in vivo. In addition, the affected cells appeared to have a reduction in mitosis and alcian blue staining proteoglycans. Cultures from affected animals contained fewer cells positive for alkaline phosphatase activity. Both affected and control cells expressed collagen types I and II and were positive for the lectin Ricinus communis agglutinin-I. However, the staining of the control culture for type II collagen was more prominent than in the affected cells. These findings suggest that culture of chondrocytes in agarose may be a useful method for studying the biology of cartilage which leads to skeletal abnormalities in lysosomal storage diseases.

Animals↗

Retarded bone formation in GM1-gangliosidosis: a study of the infantile form and comparison with two canine models.

The development of skeletal lesions in two canine models of GM1-gangliosidosis, English springer spaniels and Portuguese water dogs, has been studied and compared to osseous abnormalities in a child with the infantile form of the disease. In the canine models, skeletal dysplasia was progressive. Lesions were noted at 2 months of age and characterized by retarded endochondral ossification and osteoporosis. Older puppies had focal cartilage necrosis within lumbar vertebral epiphyses. At the cellular level, lesions were characterized by chondrocytic hypertrohy and lysosomal accumulation of storage compounds. Our studies illustrate that the skeletal lesions in both canine models are similar to those in a child with GM1-gangliosidosis. Furthermore, we proposed that the abnormal storage of partially degraded compounds in affected chondrocytes might explain, at least in part, the retarded bone formation noted in patients with GM1-gangliosidosis.

Animals↗

A light microscopical, ultrastructural and immunohistochemical study of spindle-cell adrenocortical tumours of ferrets.

Twelve adrenocortical tumours with a variable spindle-cell component in ferrets (six spayed females, three intact females, two castrated males, and one intact male) were examined by light microscopy. One tumour with a moderate spindle-cell component was examined ultrastructurally, and three tumours were studied immunohistochemically. Light microscopy revealed a spindle-cell component in the tumours that varied from a few such cells occupying the stroma between packets of adrenocortical cells to cells in such large numbers that they formed almost the entire substance of the tumour. By light microscopy these spindle cells resembled smooth muscle cells, and the ultrastructural findings, particularly the presence of thin contractile filaments, suggested that the spindle cells were of smooth muscle origin. Immunohistochemical staining revealed that the spindle cells were negative for cytokeratins and S-100 protein but positive for smooth muscle actin. Desmin was readily demonstrated in two tumours but not in the other examined. Vimentin was variable, generally producing a small to moderate amount of reaction product.

Actins↗

Vitamin D inhibits angiogenesis in transgenic murine retinoblastoma.

PURPOSE: Vitamin D compounds have been shown to inhibit tumor growth in a transgenic retinoblastoma murine model. The mechanism of action has not been defined clearly, although an antiangiogenic action has been proposed. METHODS: Transgenic retinoblastoma mice received high (0.05 microgram) and low (0.025 microgram) doses of vitamin D3 by intraperitoneal injection 5 times per week for 5 weeks. Control animals were injected with mineral oil vehicle alone. At 5 months of age, the animals were killed and eyes were enucleated and processed for light microscopy. Paraffin-embedded sections were stained with an immunoperoxidase stain (GS-1) specific for mammalian vascular endothelium. Sections were graded by a single masked reviewer, and intraobserver reliability was assessed. Mean vessel counts were made for each group. RESULTS: The high-dose group had the lowest mean vessel count (8.5), followed by the low-dose group (10.1). The control group had the highest mean vessel count (14.1). Vitamin D-treated animals (high- and low-dose groups combined) had significantly fewer vessels P = 0.001) than untreated controls. CONCLUSIONS: These results support the hypothesis that inhibition of angiogenesis is a mechanism of action for vitamin D in the transgenic retinoblastoma mouse model.

Animals↗

The effect of lysosomal storage diseases on secretory cells: an ultrastructural study of pancreas as an example.

The lysosome is the disposal and recycling center of eukaryotic cells. The pancreas is an exocrine and endocrine organ. It contains different cell types secreting enzymes, hormones, ions and fluid. The purpose of this study was to demonstrate that the manifestation of the disease in individual cells depends on the expression of the defected gene within the function of the cells. We have thus compared the ultrastructural features of normal pancreas to pancreas affected by eleven different lysosomal storage diseases in human, cats, dogs and mice. The diseases studied include GM1- and GM2- gangliosidosis, galactosialidosis, alpha-mannosidosis, mucopolysaccharidosis-I (MPS-I), mucopolysaccharidosis-VI (MPS-VI), mucopolysaccharidosis-VII (MPS-VII), mucolipidosis-IV (ML-IV), Niemann-Pick C, neuronal ceroid-lipofuscinosis, and an undefined glycolipid storage disease. We demonstrated that morphological changes are seen in some pancreatic cell types but not in others, even though the genetic defect occurs in all cell types. Furthermore, we identified and characterized primary and secondary changes in affected cells. The primary changes include presence of numerous and/or enlarged secondary lysosomes laden with partially degraded substrates. The secondary changes were associated with an abnormal formation and maturation of zymogen or secretory granules. The mechanisms contributing to the secondary changes are thought to be associated with altered synthesis of various constituents of these granules.

Animals↗

Mucolipidosis IV: morphology and histochemistry of an autopsy case.

Mucolipidosis Type IV is a rare, autosomal recessive disorder characterized by corneal opacification, mental retardation, and delayed motor milestones. Whereas lysosomal storage material has been demonstrated in biopsied tissues and leukocytes, the complete autopsy pathology, including neuropathology, is unknown. The metabolic defect remains speculative. We report the general and neuropathologic findings of the only known autopsy. In the central nervous system, neuronal loss in the cerebral cortex, basal ganglia, deep cerebellar nuclei, and brainstem nuclei was marked by astrocytosis; the cytoplasm of residual neurons had brown granules. These granules were positive with periodic acid-Schiff, Concanavalia ensiformis, and Sudan black, but not with Luxol-fast blue. Ultrastructurally, neurons contained lysosomes laden with osmiophilic, amorphous and granular material, and few lamellated membrane structures. Hepatocytes, epithelia, endothelia, chondrocytes, and tissue macrophages also stained positively with Datura stramonium and Ricinus communis-I agglutinins, with renal glomeruli also staining with peanut agglutinin; most non-neural cells contained osmiophilic granules on toluidine blue-stained, plastic embedded sections, corresponding to lamellated membrane structures. These findings complement the previously reported ocular morphology and brain and liver biochemistry performed in the same patient, and suggest that the storage material in neurons differs from that in non-neural cells. Furthermore, the underlying defect is not likely to be a deficiency of a single enzyme (i.e. a lysosomal hydrolase).

Adult↗

A novel in situ model to study Pneumocystis carinii adhesion to lung alveolar epithelial cells.

Pneumocystis carinii, an extracellular parasite thriving in the lungs of immunosuppressed mammals, is a major cause of death in AIDS patients in the USA. As a prelude to growth, the parasite adheres mostly to type I pneumocytes lining the alveolar spaces. The mechanism of adherence remains unknown, largely because of difficulties in isolating type I pneumocytes and maintaining them in vitro. As a first step to understand P. carinii adherence to its natural substrate, we developed an in situ method to directly study parasite binding to lung alveolar cells. We used formaldehyde-fixed paraffin-embedded sections of normal rat lung as substrate for adhesion. As in its binding to the lungs in vivo, P. carinii adhered preferentially to type I pneumocytes. Adherence was saturable, time and dose dependent, and selectively blocked by glycoconjugates, in particular bovine submaxillary mucin, fetuin, and asialofetuin, suggesting that it may be mediated by a lectin type of interaction. Further, IgG of rats with P. carinii pneumonia inhibited adherence, suggesting that it may react with parasite ligands involved in the recognition of type I cell receptors. Our results demonstrate the usefulness of the in situ model for studying the mechanisms of P. carinii adherence to alveolar cells. In addition, this method may be valuable for identifying neutralizing antibodies and drugs potentially useful for controlling the infection in vivo.

Animals↗

Lectin histochemistry of infantile lysosomal storage disease associated with osteopetrosis.

In infantile lysosomal storage disease associated with osteopetrosis the nature of the enzyme deficiency as well as the type of material accumulated are both unknown. We used lectin histochemistry to characterize the storage material of previously reported cases. Using paraffin sections neurons stained positively with Luxol fast blue (LFB), periodic acid-Schiff (PAS), Concanavalia ensiformis agglutinin, Datura stramonium agglutinin, Griffonia simplicifolia-I, Lens culinaris agglutinin, Ricinus communis agglutinin-I, succinylated wheat germ agglutinin and wheat germ agglutinin, indicating an accumulation of fucosylated N-glycosidically linked oligosaccharides containing beta- and alpha-galactosyl residues and compounds containing N-acetyllactosamine. Reticuloendothelial cells in liver and in spleen did not stain with LFB, but did stain with PAS and the above lectins. These results indicate that there is storage of both carbohydrates and lipids in neurons, and stored carbohydrates with similar residues in reticuloendothelial cells in this disease, where the primary defect is still unknown.

Brain↗

Developmentally regulated lectins in Eimeria species and their role in avian coccidiosis.

Avian coccidiosis caused by Eimeria species is characterized by rather specific site infections of the intestine. We used hemagglutination and hemagglutination inhibition assays on various developmental stages of Eimeria tenella and on sporozoites of Eimeria acervulina and Eimeria maxima to assay for parasite lectins. Various monosaccharides, polysaccharides, and glycoproteins were used to demonstrate differences in sugar specificity of the lectins between these species. Surface lectins were found on the primary infective stage, i.e., sporozoites, but not on merozoites or unsporulated oocysts. Also, there were differences in the specificities of the various sugar lectins among the different parasite species. Furthermore, there was a dose-dependent reduction of infection of tissue culture cells by sporozoites of E. tenella that were continuously exposed to fetuin, 1 of the specific inhibitors of the lectin. The results of our study are unique in that in 3 species of avian Eimeria all have a lectin on their sporozoites, but the lectins for each species have different sugar specificities. We hypothesize that these lectins found on the surface of the sporozoites may play a role in determining the site of infection within the intestine of the host.

Animals↗

Adult-onset lysosomal storage disease in a Schipperke dog: clinical, morphological and biochemical studies.

An adult-onset lysosomal storage disorder was diagnosed in a 5-year-old Schipperke dog with progressive cerebellar and central vestibular signs. It was characterized by cerebellar atrophy with extensive loss of Purkinje and granular cells, and hydrocephalus. Enlarged and vacuolated neurons were observed in spinal cord and brain; pancreatic centrolobular and islet cells were also vacuolated. Ultrastructurally, enlarged secondary lysosomes laden with lamellated membrane structures were present in neurons and empty enlarged vacuoles were found in pancreatic centroacinar, ductal, and islet cells. On frozen sections neurons stained with Ricinus communis agglutinin-I and wheat germ agglutinin. On paraffin sections neurons stained with luxol fast blue, periodic acid-Schiff, Concanavalia ensiformis agglutinin, and were autofluorescent. These findings indicate an accumulation of glycolipids containing terminal beta-galactosyl and alpha-sialyl residues, and N-linked oligosaccharides. Tissue activity of lysosomal beta-galactosidase was 50% of normal and the activity of beta-hexosaminidase was elevated. Brain lipid-bound sialic acid was twice normal, with a small increase of GM1-ganglioside, but there was a significant elevation of GM2 (GD2) and GM3 (GD3). In addition, significant elevations of sialylated and non-sialylated oligosaccharides were noted. These clinical, biochemical and pathological findings are similar to those observed in human patients with adult-onset galactosialidosis.

Animals↗

Identification of a developmentally regulated sialidase in Eimeria tenella that is immunologically related to the Trypanosoma cruzi enzyme.

Sporozoites and merozoites of three species of Eimeria, E. tenella, E. maxima, and E. necatrix, that cause diarrhea in chickens worldwide, were examined for their expression of sialidase (SA) activity. The enzyme was found in three species, and the activity of merozoites was 10-20 times higher than that of sporozoites. The enzyme was resistant to degradation by proteases that are normally present in the intestine, a site inhabited by the Eimeria parasites, and it was relatively resistant to heat, with optimum activity being at 40 degrees C, which is within the range of temperature in the chicken intestine (40-43 degrees C). E. tenella SA was immunoprecipitated by monoclonal and polyclonal antibodies raised against the Trypanosoma cruzi SA (TCSA), and enzyme activity was neutralized by these antibodies. E. tenella SA was identified by immunoblots as a doublet of molecular weight 190,000 and 180,000 using, as a probe, anti-TCSA antibodies and antibodies against a synthetic peptide (TR) derived from the long tandem repeat domain of TCSA. Binding of the monoclonal and polyclonal antibodies to E. tenella was completely blocked by TR, but not by an irrelevant peptide (BR). Therefore, E. tenella expresses a developmentally regulated SA that is structurally related to the T. cruzi counterpart. Because of the high SA activity in merozoites, and by analogy with other SA-producing microbes that inhabit mucin-rich epithelia, we suggest that the Eimeria SA plays a role in desialylating intestinal mucins to reduce viscosity of the local environment and thereby facilitate parasite migration. The enzyme could also play a role in host cell-parasite interaction.

Amino Acid Sequence↗

Ultrastructure of human dermal mast cells in 29 different lysosomal storage diseases.

The effect of lysosomal storage diseases on the ultrastructure of human mast cells has not previously been reported. Indeed, there has been little published evidence indicating that mast cells contain typical lysosomes. However, mast cell cytoplasmic granules contain hydrolases similar to those found in lysosomes, but which differ from lysosomal hydrolases in exhibiting optimal activity at higher pH. We therefore examined by transmission electron microscopy the dermal mast cells in 58 biopsies of patients exhibiting 1 of 29 different lysosomal storage diseases. We found mast cells containing abnormal lysosomes in 16 of these disorders. In 6 of these 16 diseases, the mast cells' cytoplasmic granules appeared normal. These observations indicate that human mast cells can contain lysosomes, and provide evidence that the enzymes affected by lysosomal storage diseases are active in mast cells.

Biopsy↗

Purification and immunological characterization of acid beta-galactosidase from dog liver.

1. Dog liver acid beta-galactosidase was isolated in high yield and purified to homogeneity using a series of chromatographies on Con A-Sepharose, decyl-agarose, anion-exchange HPLC and gel-filtration HPLC. 2. Non-denaturing gel filtration by HPLC gave a single homogeneous peak corresponding to molecular mass of 180-190 kDa. During SDS-PAGE analysis, the single peak dissociated into a major band corresponding to molecular mass of 32 kDa with minor bands at 18 and 13 kDa. 3. Polyclonal antibodies raised against the purified enzyme immunoprecipitated beta-galactosidase activity specifically from dog liver extracts and recognized a single 32 kDa band in Western blot analysis of dog tissue homogenates. This antibody did not crossreact with any protein band in tissue homogenates from other species examined except cat. 4. Western blot analysis of tissue extracts from dogs affected with GM1-gangliosidosis showed the presence of a 32 kDa band similar to that of controls.

Animals↗

Myositis due to Pleistophora (Microsporidia) in a patient with AIDS.

Microsporidia are obligate intracellular protozoa that parasitize both vertebrates and invertebrates and are now recognized as important pathogens in individuals infected with human immunodeficiency virus type 1 (HIV-1). We describe the clinical and morphological features of a case of pleistophora (microsporidian) myositis in a patient with AIDS and delineate the stages of the microsporidian life cycle, as visualized by light and electron microscopy. We discuss significant aspects of microsporidian infections in humans and of myopathy attributable to other causes in HIV-1-infected individuals. As far as we know, ours is only the second reported case of microsporidian myositis and the first reported case in a patient with documented HIV-1 infection.

AIDS-Related Opportunistic Infections↗

The role of the eae gene of enterohemorrhagic Escherichia coli in intimate attachment in vitro and in a porcine model.

The eaeA gene of enteropathogenic Escherichia coli (EPEC) is necessary for intimate attachment to epithelial cells in vitro. Enterohemorrhagic E. coli (EHEC) strains also possess an eae gene and are capable of intimate attachment and microvillus effacement in vitro and in animal models. To assess the role of the EHEC eae gene in intimate attachment, we constructed an eae deletion/insertion mutation in wild-type EHEC O157:H7 strain 86-24 by using linear electroporation of a recombinant allele. The mutant obtained was deficient in inducing f-actin accumulation in HEp-2 cells and was incapable of attaching intimately to colonic epithelial cells in a newborn piglet model of infection. Intimate attachment in vivo was restored when the EHEC eae gene or the eaeA gene of EPEC was introduced into the mutant on a plasmid. These results indicate that the eae gene is necessary for intimate attachment of EHEC in vivo. In addition, the complementation achieved by the EPEC locus indicates that the eae gene of EHEC and the eaeA gene of EPEC are functionally homologous.

Adhesins, Bacterial↗