Extraskeletal Osteosarcoma of the Thigh: An Autopsy Case Report

We report a case of extraskeletal osteosarcoma (ESOS) and autopsy findings. A 35-year-old man presented with an ossified tumor in the right thigh and lung metastasis. The lung tumors continued to develop despite multiagent chemotherapy and caused death within 8 months. Autopsy revealed many secondary lesions in the lungs, especially in the left lung. Histopathologically, the primary tumor and one of the secondary tumors showed proliferation of spindle-shaped tumor cells focally forming lace-like osteoid material. Therefore, we made a definite diagnosis of ESOS

Porcine dentin sialoprotein glycosylation and glycosaminoglycan attachments

<p>Abstract</p> <p>Background</p> <p>Dentin sialophosphoprotein (Dspp) is a multidomain, secreted protein that is critical for the formation of tooth dentin. Mutations in <it>DSPP </it>cause inherited dentin defects categorized as dentin dysplasia type II and dentinogenesis imperfecta type II and type III. Dentin sialoprotein (Dsp), the N-terminal domain of dentin sialophosphoprotein (Dspp), is a highly glycosylated proteoglycan, but little is known about the number, character, and attachment sites of its carbohydrate moieties.</p> <p>Results</p> <p>To identify its carbohydrate attachment sites we isolated Dsp from developing porcine molars and digested it with endoproteinase Glu-C or pronase, fractionated the digestion products, identified fractions containing glycosylated peptides using a phenol sulfuric acid assay, and characterized the glycopeptides by N-terminal sequencing, amino acid analyses, or LC/MSMS. To determine the average number of sialic acid attachments per N-glycosylation, we digested Dsp with glycopeptidase A, labeled the released N-glycosylations with 2-aminobenzoic acid, and quantified the moles of released glycosylations by comparison to labeled standards of known concentration. Sialic acid was released by sialidase digestion and quantified by measuring β-NADH reduction of pyruvic acid, which was generated stoichiometrically from sialic acid by aldolase. To determine its forms, sialic acid released by sialidase digestion was labeled with 1,2-diamino-4,5-methyleneoxybenzene (DMB) and compared to a DMB-labeled sialic acid reference panel by RP-HPLC. To determine the composition of Dsp glycosaminoglycan (GAG) attachments, we digested Dsp with chondroitinase ABC and compared the chromotagraphic profiles of the released disaccharides to commercial standards. N-glycosylations were identified at Asn³⁷, Asn⁷⁷, Asn¹³⁶, Asn¹⁵⁵, Asn¹⁶¹, and Asn¹⁷⁶. Dsp averages one sialic acid per N-glycosylation, which is always in the form of N-acetylneuraminic acid. O-glycosylations were tentatively assigned at Thr²⁰⁰, Thr²¹⁶and Thr³¹⁶. Porcine Dsp GAG attachments were found at Ser²³⁸and Ser²⁵⁰and were comprised of chondroitin 6-sulfate and chondroitin 4-sulfate in a ratio of 7 to 3, respectively.</p> <p>Conclusions</p> <p>The distribution of porcine Dsp posttranslational modifications indicate that porcine Dsp has an N-terminal domain with at least six N-glycosylations and a C-terminal domain with two GAG attachments and at least two O-glycosylations.</p

Porcine Enamel Protein Fractions Contain Transforming Growth Factor‐β1

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141064/1/jper1688.pd

Modification of intracellular Ca2+ dynamics by laser inactivation of inositol 1,4,5trisphosphate receptor using membrane-permeant probes. Chem Biol. 2004; 11:1053–1058. [PubMed: 15324806

studying of the role of the protein in these functions. We developed a chromophore-labeled IP 3 analog (carboxymalachite green-aminopropyl-1D-myo-inositol-1,4,5-Introduction trisphosphate, MGIP 3 ) and showed that spatially controlled inactivation of IP 3 R could be achieved within a Specific inactivation of biomolecules has been one of few seconds by MGIP 3 -mediated smCALI [8-10]. The the most widely used approaches to study the physiotime required for smCALI is very much shorter than that logical functions of these molecules, and various methof antibody-based CALI, which usually requires ‫5ف‬ min. ods, including the use of pharmacological antagonists, Furthermore, by focusing a laser beam at the subcellular targeted gene disruption, and antibodies, have been level in a single PC12 cell, precise control of the area used. The activities of most functional biomolecules deof inactivation could be achieved. In the present study, pend upon the site of expression and the nature of the we derivatized MGIP 3 to a membrane-permeant comcellular processes in which they are involved [1-3], so pound for noninvasive delivery into living cells. This a technique for inactivation of target proteins in a spatiomembrane-permeant probe can be easily applied to intemporally well controlled manner should be extremely tact cells or cell populations and smCALI experiments useful. To achieve this purpose, chromophore-assisted can be conducted under physiological conditions. We laser inactivation (CALI) is an excellent method [4, 5], in describe here the successful application of smCALI in which the protein of interest is targeted by an exogeintact cells using the membrane-permeant probe, and nously introduced antibody that has been tagged with we show that smCALI provides a powerful tool to anaa CALI chromophore, usually malachite green, and the lyze complex intracellular Ca 2ϩ dynamics. chromophore-antibody complex is subsequently irradiated with intense localized laser light. The irradiated Results and Discussion chromophore produces radical species that react with the protein to which the antibody is bound, causing MGIP 3 /PM Permeates through the Plasma its inactivation. Because the radical species are highly Membrane and Interacts with IP 3 R In previous studies, we designed and synthesized a chromophore-labeled IP 3 analog (MGIP 3