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PubMed1991

Ubiquitin and heat shock protein expression in amyotrophic lateral sclerosis.

The expression of two heat shock proteins, HSP72 and p57, in addition to ubiquitin, has been studied immunocytochemically in nine amyotrophic lateral sclerosis (ALS) cases and 10 age-matched controls. HSP72 and p57 antibodies did not identify the characteristic ubiquitin-immunoreactive inclusions present in anterior horn cells in ALS spinal cord. Antibodies to HSP72, but not to p57 or ubiquitin, strongly labelled structures corresponding to polyglucosan bodies in spinal grey matter. Such immunoreactive profiles were more abundant in ALS cases, although they were also present in control material. They were sometimes identified by haematoxylin and eosin and periodic acid Schiff reaction, but were not labeled by phosphotungstic acid haematoxylin or by antibodies to glial fibrillary acidic protein. Although ubiquitin, HSP72 and p57 are stress-induced proteins, they are expressed differently and might therefore have different significance in neuronal degeneration.

Aged↗

PubMed1996

Oncocytic hepatocellular carcinoma with numerous globular hyaline bodies.

Two well circumscribed tumors, oncocytic and non-oncocytic, were removed from the non-cirrhotic liver of a 67 year old male. The large oncocytic tumor (OCT), occupying the entire left lobe, was multilobulated with focal coagulation necrosis and areas of hemorrhage. Light microscopy revealed that it consisted of exclusively large, granular oxyphilic cells with moderate nuclear atypia and occasional mitotic figures, which were trabecular and/or pseudoglandular in structure, but no lamellar fibrosis was seen. Characteristically, the OCT cells included numerous globular hyaline bodies (GHB) of various sizes which were stained red with acid fuchsin and deep blue or magenta with phosphotungstic acid hematoxylin (PTAH), but negative for periodic acid Schiff (PAS), orcein, rhodamine and Grimelius methods. Immunohistochemically, alpha-fetoprotein (AFP), alpha-1-antitrypsin, alpha-1-antichymotrypsin, fibrinogen and ferritin were all negative. On ultrastructural examination, tumor cells were mitochondria-rich, including electron dense, ovoid or polyhedral inclusions, with the delineated membrane identical with that of the GHB. In contrast, the small tumor in the right lobe (Segment 7) was a solid adenoma with no oncocytic transition. Based on these findings, it was postulated that OCT consists of heterogenous proliferation of mitochondria-rich hepatocytes which tend to induce lysosomal GHB closely associated with mitochondrial abnormalities.

Adenocarcinoma↗

PubMed1972

Purification and characterization of epizootic diarrhea of infant mice virus.

Epizootic diarrhea of infant mice virus has been purified from the intestines of infected mice using enzymatic digestion, precipitation with polyethylene glycol, extraction with Genesolv-D, and sucrose step-gradient centrifugation. The purified virus was found to be stable at -70 C or when treated with ether, chloroform, or sodium deoxycholate. Biochemical analysis of purified virions has suggested that the nucleic acid type of epizootic diarrhea of infant mice virus was of the ribonucleic acid type. Preparations of purified virions stained with phosphotungstic acid showed a particle with icosahedral symmetry, 54 +/- 2 nm in diameter and probably composed of 32 hollow capsomeres.

Animals↗

PubMed1997

Pathology of idiopathic megarectum and megacolon.

BACKGROUND: The aetiology and pathology of both idiopathic megarectum and idiopathic megacolon are unknown. In particular, it is unknown whether there are abnormalities involving enteric nerves or smooth muscle. METHODS: Resected tissue was examined from 24 patients who underwent surgery for idiopathic megarectum, from six patients who had tissue resected for idiopathic megacolon, and 17 control patients who had surgery for non-obstructing large bowel cancer. Qualitative and quantitative histological examination was performed after staining with haematoxylin and eosin, periodic acid Schiff (PAS), Martius scarlet blue (MSB), and phosphotungstic acid haematoxylin (PTAH). Neural and glial tissue were examined after immuno-staining with S100 and PGP9.5. RESULTS: Compared with controls, patients with idiopathic megarectum had significant thickening of their muscularis mucosae (median 78 v 33 microns, p < 0.005), circular muscle (1000 v 633 microns, p < 0.005), and longitudinal muscle (1083 v 303 microns, p < 0.005), despite rectal dilatation. This thickening was relatively greater in the longitudinal than in the circular muscle. Fibrosis of the longitudinal muscle was seen, using MSB staining, in 58%, of circular muscle in 38%, and of muscularis mucosae in 29% of patients. The relation between muscle thickening and fibrosis was variable. The density of neural tissue in the longitudinal muscle seemed to be reduced in patients with idiopathic megarectum. There was no thickening of enteric muscle or alteration in the density of innervation in patients with idiopathic megacolon. CONCLUSION: There is notable thickening of the enteric smooth muscle in patients with idiopathic megarectum, but the architecture of the enteric innervation seems to be intact. Functional abnormalities of the latter remain a possible cause of the smooth muscle hypertrophy.

Adolescent↗

PubMed1990

Neoplastic Paneth cells in the experimental murine carcinoma of the small intestine.

The purpose of this study is to elucidate the participation of Paneth cells in experimentally induced adenocarcinoma of the intestine. The rats were fed with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) dissolved in drinking water ad libitum at a concentration of 100 micrograms/ml for 28 weeks. They were sacrificed 12 weeks after the last MNNG administration. A number of tumor cells containing large eosinophilic granules in their supranuclear cytoplasm (Paneth cells) were observed in about 20% of the experimentally induced adenocarcinoma of the small intestine. The granules were stained positively with Lendrum, periodic acid-Schiff, Masson's trichrome, and Mallory's phosphotungstic acid hematoxylin. Ultrastructurally, the granules were round, osmiophilic, and relatively even in size. We compared the morphologic features of the Paneth cell-containing small intestinal adenocarcinomas (Group I) with those without Paneth cells (Group II). Group I was distinguished from Group II by its better differentiation, larger tumor size and lower incidence of calcification. Although Paneth cells are extremely rare in human gastrointestinal carcinomas, twenty percent of MNNG-induced intestinal carcinomas harbor Paneth cells. The neoplastic Paneth cells in experimental carcinomas may differentiate from uncommitted cells in the deeper portion of the crypt.

Adenocarcinoma↗

PubMed1978

Thin layer chromatographic method for the detection of uric acid: collaborative study.

A collaborative study has been completed on an improved method for the detection and confirmation of uric acid from bird and insect excreta. The proposed method involves the lithium carbonate solubilization of the suspect excreta material, followed by butanol-methanol-water-acetic acid thin layer chromatography, and trisodium phosphate-phosphotungstic acid color development. The collaborative tests resulted in 100% detection of uric acid standard at the 50 ng level and 75% detection at the 20-25 ng level. No false positives were reported during tests of compounds similar to uric acid. The proposed method has been adopted official first action; the present official final action method, 44.161, will be retained for screening purposes.

Animals↗

PubMed2006

Effect and mechanism of total saponin of Dioscorea on animal experimental hyperuricemia.

In this study, we investigated the effects and mechanisms of Total Saponin of Dioscorea (TSD) on animal experimental hyperuricemia. Mouse and rat hyperuricemic models were made by orally administering yeast extract paste once a day (30 and 20 g/kg, respectively), for 7 days. Yeast would disturb normal purine metabolism by increasing xanthine oxidase (XOD) activity and generating large quantities of uric acid. This model is similar to human hyperuricemia, which is induced by high-protein diets, due to a purine and nucleic acid metabolic disturbance. Another mouse hyperuricemia model was generated by intraperitoneal injection once with uric acid 250 mg/kg or potassium oxonate 300 mg/kg. Potassium oxonate, a urate oxidase inhibitor, can raise the serum uric acid level by inhibiting the decomposition of uric acid. Likewise, injecting uric acid can also increase serum uric acid concentration. The concentration of uric acid in serum or urine was detected by the phosphotungstic acid method, and the activity of XOD was assayed by a test kit. The results showed that TSD (240, 120 and 60 mg/kg, ig) could significantly lower the level of serum uric acid in hyperuricemic mice. TSD (120 and 60 mg/kg, ig) could also lower the level of serum uric acid in hyperuricemic rats, reduce the activity of XOD in the serum and liver of hyperuricemic rats, and increase the level of urine uric acid concentration as well as 24-hour total uric acid excretion. In conclusion, TSD possesses a potent anti-hyperuricemic effect on hyperuricemic animals, and the mechanism may be relevant in accelerating the excretion and decreasing the production of uric acid.

Animals↗

PubMed1991

A cytochemical and immunocytochemical study of DNA distribution in spermatid nuclei of mouse, rabbit, and bull.

DNA distribution in mouse, rabbit and bull spermatids was analyzed by electron microscopy, after using a Feulgen-like HCl-osmium ammine procedure, and after immunocytochemistry with anti-DNA antibodies. In addition, nucleic acids were visualized with the intercalating dye ethidium bromide and phosphotungstic acid. The parts of DNA displaying a beta helix configuration (possibly A-T rich parts) were identified by epifluorescence microscopy after staining with Hoechst 33258. In all 3 species, young spermatid nuclei were seen to have large areas poor in DNA, as well as DNA-rich areas, which were mostly concentrated into a peripheral layer close to the acrosome and into one or several masses, displaying species-specific locations. These DNA-rich areas were stained with Hoechst 33258. Elongating spermatid nucleic contained homogeneously distributed DNA, and this was evident following both immunocytochemistry and nucleic acid histochemistry in all 3 species. However, the distribution appeared more heterogeneous after the Feulgen-like procedure, and was accompanied by a disappearance of Hoechst-fluorescence. In fully elongated spermatids, all nuclear areas stained with Hoechst 33258, while the 3 other techniques labeled either all or species-specific parts of the condensed chromatin. The reasons for these variable reactions are discussed in terms of technique specificities, DNA configuration and nucleoprotein moiety replacements.

Animals↗

PubMed1991

Predominant Paneth cell differentiation in an intestinal type gastric cancer.

A distinct morphological variant of a diffuse type adenocarcinoma of the stomach with Paneth cell differentiation is reported. The tumor was a Borrmann's Type III carcinoma measuring 6.0 x 5.5 cm at the body along the greater curvature. It was composed of Paneth cell- and endocrine cell differentiated cancer cells in addition to tubular and poorly differentiated adenocarcinoma cells. The Paneth cell differentiation was characterized histologically by cytoplasmic distinct coarse eosinophilic granules stained red with periodic acid-Schiff and Masson trichrome reagents and reddish brown with phosphotungstic acid hematoxylin, and electron microscopically by lysozyme in cytoplasmic electron dense granules. In addition, electron microscopy revealed acid mucin globules and various intermediate forms between Paneth granules and the mucin globules which might be regarded as abortive forms of Paneth granules presumably resulting from defective incorporation of lysozyme-positive mucosubstances into acid mucin. Endocrine differentiated cancer cells consisted of serotonin-, peptide YY-, and glucagon/glicentin-positive cells. The various cell phenotypes found in the present tumor could be explained on the basis of intestinal differentiation of gastric cancer.

Adenocarcinoma↗

PubMed1984

Ultrastructural characterization of normal and damaged membranes of Mycobacterium leprae and of cultivable mycobacteria.

Microdensitometry showed that the membrane profiles of normal cultivable mycobacteria were very asymmetric (outer layer denser and thicker than the inner layer), while the profiles of normal-looking M. leprae in lepromatous patients, in experimentally infected armadillos and in nude mice were approximately symmetric; moreover, the membrane of M. leprae was thicker than that of cultivable species. Using two cytochemical methods for the ultrastructural detection of periodic acid-Schiff (PAS)-positive molecules (the Thiéry procedure, and staining with phosphotungstic acid at low pH) we found that the membrane of cultivable mycobacteria, growing in vitro or in vivo, had PAS-positive components exclusively in the outer layer, while the normal-looking M. leprae in patients and in armadillos had membranes with PAS-positive components in both layers. The membranes of damaged cultivable mycobacteria, in vivo or in vitro, and of damaged M. leprae, in patients or armadillos, were PAS-negative.

Animals↗

PubMed2005

Primary extraneural myxopapillary ependymoma of the broad ligament.

Primary extraneural ependymomas are rare tumors that arise in ectopic sites, including pulmonary, sacrococcygeal region, ovarian, and paraovarian tissues. Four such ependymomas reported in the literature involve the paraovarian tissues, including 2 broad ligament ependymomas. Here we describe a myxopapillary ependymoma of the broad ligament in a 22-year-old woman, which may be the first tumor of this type to be reported in this location. Cytology, histology, cytochemistry, immunohistochemistry, and flow cytometry ploidy analysis are studied and described. Identification of perivascular ependymal rosettes, ependymal canals, vimentin and glial fibrillary acidic protein immunoreactivity, cytochemical staining of blepharoplasts or terminal bars by phosphotungstic acid hematoxylin, and presence of multiple foci of myxoid degeneration among the ependymal rosettes characterized a myxopapillary ependymoma.

Adult↗

PubMed1990

Determination of uric acid in canine serum and urine by high performance liquid chromatography.

A reproducible high performance liquid chromatography (HPLC) method was developed for analysis of uric acid in canine serum and urine. The method consists of precipitating serum proteins with phosphotungstic acid prior to HPLC analysis. Urine is analyzed after dilution with buffer. Chromatography is performed on a reversed-phase C-18 column with UV detection at 292 nm. Sensitivity of the method will allow reproducible measurement of uric acid at concentrations of 0.05 mg/dl in serum and 0.1 mg/dl in urine. The HPLC method has been used to quantify hundreds of canine serum and urine samples. The method is superior to UV absorption or colorimetric methods because its lower limit of detection allows measurement of uric acid at concentrations found in canine serum and urine.

Journal Article↗

PubMed1991

[Sialyltransferase activity in the spermatic membrane and its relation to human fertility and sterility].

Sialyltransferase activity was determined on normospermic men sperm cells (greater than 80 x 10(6) sperm/ml and 75% motility) and oligospermic infertile sperm cells (less than 20 x 10(6) sperm/ml and less than 20% motility) and asthenospermic (greater than 40 x 10(6) sperm/ml and less than 10% motility). Sialytransferase activity is quantified by means of the transference of radioactivity of CMP-3H-sialic acid toward the exogenous acceptor (asialofetuin). The enzyme substrate complexes formed in presence of phosphotungstic acid result precipitated insoluble, which was retained on glass fiber filter. The sialyltransferase activity decrease in oligospermic sperm cells 62 +/- 3% and in the asthenospermic decreased 57 +/- 4% with respect normospermic sperm cells. The decrement on sialyltransferase activity in the infertile sperm, permits to assume that this enzyme probably participates as a direct cause of its pathology with detrimental structural and functional integrity of the plasma membrane.

Cell Membrane↗

PubMed1966

Structure of Bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid.

Anderson, D. L. (University of Minnesota, Minneapolis), D. D. Hickman, and B. E. Reilly. Structure of Bacillus subtilis bacteriophage phi29 and the length of phi29 deoxyribonucleic acid. J. Bacteriol. 91:2081-2089. 1966-Bacillus subtilis bacteriophage phi29 were negatively stained with phosphotungstic acid. The head of phi29 has a hexagonal outline with a flattened base, and is about 315 A wide and 415 A in length. The virus has an intricate tail about 325 A in length. Twelve spindle-shaped appendages are attached to the lower of two collars which comprise the proximal portion of the tail. The distal 130 A of the tail axis has a diameter of about 60 A and is larger in diameter than the axis of the upper portion of the tail. Comparison of electron microscopic counts of phi29 with plaque-forming units indicated that about 50% of the microscopic entities were infective. Phenol-extracted phi29 deoxyribonucleic acid (DNA) molecules were prepared for electron microscopy by the cytochrome c film technique of Kleinschmidt et al. Measurement of contour lengths of DNA molecules from three preparations gave skewed distributions of lengths with observed modal class values ranging from 5.7 to 5.9 mu. Assuming that phi29 DNA is a double helix in the B form, the corresponding molecular weights would be 10.9 x 10(6) to 11.3 x 10(6) daltons. The largest DNA molecules would have a volume of 1.9 x 10(7) A(3) which is about 25% greater than the estimated 1.4 x 10(7) A(3) internal volume of the phage head.

Bacillus subtilis↗

PubMed1976

[Modification of the Romeis method for cartilage staining].

When staining of the cartilage by the method of Romeis one can increase of the electivity using more acidic staining solution for a shorter period. For fixation instead of solution of ammoniummolibdate, solution of phosphotungstic acid is recommended.

Animals↗

PubMed1990

Primary glomerular fibrosis: a new nephropathy caused by diffuse intra-glomerular increase in atypical type III collagen fibers.

A 38-year-old Japanese man suffering from nephrotic syndrome and slowly progressive impairment of renal function underwent renal biopsy. Light microscopy revealed diffuse lobular accentuation of glomerular tufts, occupied by homogeneous material which was eosinophilic, lightly stained with periodic acid silver methenamine and deeply with aniline blue. Further examinations using an electron microscope after treatment with phosphotungstic acid revealed that this material exhibited a diffuse increase in fibers having a periodicity of 65 +/- 4 nm (mean +/- SD) like type III collagen fibers and was reactive to the anti-type III collagen monoclonal antibody. In addition, the serum level of procollagen III peptides was markedly elevated to 221 ng/dl (normal range 2-13 ng/dl). However, the fibers were peculiarly curved and frayed as reported in the nail-patella syndrome, but were not observed in the lamina densa of the glomerular basement membrane (GBM) as seen in this syndrome. No family history or physical findings of bony and nail dystrophy, no laboratory data or pathological findings compatible with already described renal diseases including mesangiocapillary glomerulonephritis, light chain disease, immunotactoid nephropathy, amyloidosis, and non-amyloidotic fibrillary glomerulonephritis were observed. These data strongly suggest that the nephropathy is caused by a primary increase in atypical type III collagen fibers in glomeruli (primary glomerular fibrosis), which to our knowledge has not yet been described in the English literature.

Adult↗

PubMed1994

Localization of basic and acidic argyrophilic proteins in nuclear structures of elongating mouse spermatids.

Different cytochemical techniques at the electron microscopic level were used to investigate the presence of basic and acidic proteins during spermiogenesis in mice. Basic lysine rich proteins were demonstrated with an ethanolic solution of phosphotungstic acid, acidic agryrophilic proteins using two silver staining procedures applied en bloc and on the ultrathin lowicryl sections. Both types of proteins had very similar localization in the nucleoplasm and identical distribution within the postacrosomal dense lamina on the surface of late spermatid heads. The specificity of reactions is briefly discussed.

Acids↗

PubMed1993

Correlation between amino acid composition and ultrastructural features of type I and type II native collagen fibrils.

The D-band patterns of native collagen fibrils negatively stained with phosphotungstic acid (PTA) of type II (bovine cartilaginous nasal septum) and type I (bovine reticular dermis) were compared in order to find differences and correlate these with the different amino acid composition of these two collagen types. By analyzing the averaged microdensitograms of the two types of D-banding, differences in type II fibrils were observed: a) the overlap zone was more electron-transparent on average; b) there was a different hierarchy of positive peaks (intraperiod dark bands) b1, b2 and c1; c) a more marked negative peak (intraperiod light band X3) corresponded to the C-terminal extrahelical region; d) there was a different D-location of one negative peak (intraperiod light band Y3). At the same time, different values were calculated for type I and type II tropocollagen molecule total amino acid "bulkiness" (molecular volume/length ratio) and "hydrophobicity", on the basis of literature-reported data. Consistency was found between these values and the differences of the two types of D-banding. The assumption of a correlation between the above two amino acid parameters and negative staining band patterns was supported.

Amino Acids↗