Yoshio Mii

Vascularized fibular graft for reconstruction of the wrist after excision of giant cell tumor.

&NA; Seven patients with a giant‐cell tumor involving the distal end of the radius were treated with en bloc resection and reconstruction with a free vascularized fibular graft. Two patients with stage 2 disease of Ennekings surgical staging and grade 2 of Campanaccis radiographic grading system were reconstructed with an articular fibular head graft. Five patients with stage 3 and grade 3 disease underwent wrist arthrodesis using fibular shaft transfer. There was radiographic evidence of bone union at the host‐graft junctions in all cases. No local recurrence was seen in any of the patients at the most recent follow‐up examinations. There were six good and one excellent functional results. Wrist arthroplasty using a vascularized fibula head graft is the best procedure for a stage 2 or grade 2 giant‐cell tumor of the distal end of the radius. In cases of stage 3 or grade 3 disease, wrist arthrodesis using a vascularized fibular shaft graft is indicated.

Electron microscopic evidence of a viral nature for osteoclast inclusions in Paget's disease of bone

Circumstantial evidence from electron microscopic and immunological studies support the view that Pagets disease of bone represents a slow virus infection. However, there is only limited information available regarding its electron microscopic, enzyme and immunocytochemical characteristics. Two cases were studied using electron microscopy with particular emphasis on the inclusions in osteoclasts. Detailed ultrastructural and cytochemical studies including immuno-electron microscopy were performed. Some osteoclasts demonstrated specific virus-like structures composed of aggregations of microtubules in the nucleus and cytoplasm. The structures were easily digested by trypsin or protease, and were sensitive to RNase, which provided substantial evidence of a proteinaceous nature and inclusion of ribonucleic acid. Immunocytochemical examination identified binding of anti-respiratory syncytial virus and anti-measles virus antibodies in the tissue obtained from one of the two cases examined. The presence of viral antigens in structures in the cytoplasm of Pagetic osteoclasts supports the theory of paramyxovirus involvement in this disease.

The pedicled venous flap. An experimental study

Musculocutaneous pedicled venous flaps on the dorsum of rats survived at a statistically significantly higher rate than musculocutaneous composite grafts (p < 0.01). With a Silastic sheet (Dow Corning) beneath, both composite grafts and pedicled venous flaps necrosed. When a Silastic sheet with holes in it to allow some revascularisation from the bed was placed beneath, the survival rate was significantly better than with a complete Silastic sheet (p < 0.01). These results demonstrate that pedicled venous flap survival depends both on the draining vein, and revascularisation from the underlying bed.

Expression of the p16INK4a gene and methylation pattern of CpG sites in the promoter region in rat tumor cell lines

Loss of p16INK4a protein expression has frequently been related to DNA methylation in association with gene silencing. Although the methylation status of exon1α for p16INK4a involvement in various cancers has been extensively analyzed, it has been pointed out that some inconsistencies existed in its relationship to gene silencing of p16INK4a. In this study, we focused on the expression and methylation status in the regions of nt −478 to −201, containing a putative TATA box (nt −401 to −396), and nt −233 to 26, both in a recently cloned 5′ upstream region of rat p16INK4a. We showed that rat lung adenocarcinoma RLCNR did not express the p16INK4a gene, whereas rat osteosarcoma COS1NR and malignant fibrous histiocytoma MFH1NR both expressed it at levels similar to normal fibroblasts, even though the region of nt −233 to 26 was hypermethylated in COS1NR rather than RLCNR. In contrast, the CpG islands near the putative TATA box region were consistently methylated in RLCNR, but not in COS1NR and MFH1NR, as well as in normal fibroblasts. Treatment with 5‐aza 2′‐deoxycytidine induced expression of p16INK4a gene in RLCNR after 48 h, but no changes were observed in COS1NR and MFH1NR. The results indicated that methylation of CpG islands near a TATA box region played a critical role for gene silencing of the rat p16INK4a gene, rather than that of other regions.

Heterogeneous pattern of gene expression in cloned cell lines established from a rat transplantable osteosarcoma lung metastatic nodule

We have established three cloned cell lines (COS1NR, COS2NR and COS4NR) from the lung metastatic nodule of a highly metastatic variant of rat transplantable osteosarcoma, C-SLM. All three clones shared the same morphological characteristics and tumorigenicity, but their growth rates in vitro and metastatic ability in vivo differed from each other. Single-strand conformation polymorphism (SSCP) analysis revealed all three clones to have the same p53 gene mutation and parent C-SLM tumor. On the other hand, Northern blot analysis showed a different pattern of expression for the genes, c-fos, c-jun, c-Ha-ras, transin (rat stromelysin), bone Gla protein (osteocalsin) and nm23/NDP kinase. These results indicate the presence of a heterogeneous cell population in terms of the different pattern of gene expression in a lung metastatic nodule of rat osteosarcoma and the present newly established cell lines will be useful for further investigation of the biological behavior of osteosarcomas.

Ultrastructural lipid and glycoconjugate cytochemistry of membranous lipodystrophy (Nasu-Hakola disease).

In order to assess the lipid and glycoconjugate characteristics of membranous lipodystrophy, a 29-year-old male with this disease was studied using an ultrastructural cytochemical approach. The specific membranocystic lesions of the disease are composed of cystic spaces and the lining membranes. The membranes were observed to have a two-layered character: microtubular structures in the layer adjacent to the spaces and a central amorphous zone. Lipid staining and the lipase digestion test revealed triglycerides localized not only in the cystic spaces but also in the microtubular structures. Lectin histochemical examintion of carbohydrate components demonstrated thatMaclura pomifera agglutinin bound strongly to the membranes, whileGriffonia simplicifolia I,G. simplicifolia II,Concanavalia ensiformis andTriticum vulgaris agglutinin reacted weakly. Our results indicate the presence of triglycerides and carbohydrates with mainlyα-D-galactose residues in the distinctive membranocystic lesions, in particular in the microtubular structures.

Osteoclast Origin of Giant Cells in Giant Cell Tumors of Bone: Ultrastructural and Cytochemical Study of Six Cases

To clarify the histogenesis of giant cells appearing in giant cell tumors of bone (GCTB), an ultrastructural and cytochemical study of six cases was performed with both acid phosphatase (ACPase) and tartrate-resistant acid phosphatase (TRACPase) as marker enzymes. TRACPase is considered a specific marker for osteoclasts. ACPase was demonstrated in the macrophagelike stromal cells, the multinucleated giant cells, and the infiltrating macrophages. The enzyme reaction was localized in lysosomal dense bodies and Golgi areas. Intense TRACPase activity was demonstrated in the multinucleated giant cells, whereas a weak reaction was found in the macrophagelike stromal cells. The multinucleated giant cells and macrophagelike stromal cells resembled osteoclasts with regard to the subcellular localization of TRACPase. The present results suggest that the giant cells in GCTB are indeed derived from osteoclasts.

Telomerase activity correlates with growth of transplantable osteosarcomas in rats treated with cis-diammine dichloroplatinum or the angiogenesis inhibitor AGM-1470

To determine the role of telomerase activity in the growth of tumors in rats undergoing chemotherapy, a comparison of the volumes of telomerase‐positive transplantable osteosarcomas was made in rats treated with the antineoplastic agent cis‐diammine dichloroplatinum (CDDP) or the angiogenesis inhibitor O‐(chloroacetylcarbamoyl)fumagillol (AGM‐1470). Male F344 rats, 8 weeks old, received transplants of macroscopic lung metastatic nodules into the subcutaneous back space and treatment was started on day 14 thereafter. CDDP was injected i.v. at doses of 0, 0.625, 1.25 and 2.5 mg/kg body weight (b.w.) and AGM‐1470 was administered at total doses of 0, 2.5, 5 and 10 mg/kg b.w. over 2 weeks by osmotic pumps, also implanted into the subcutaneous back space, but remote from the transplanted tumors. On day 28, all animals were killed for measurement of transplanted tumor size and determination of telomerase activities by telomeric repeat amplification protocol (TRAP) assay. The results showed telomerase activity to be highly correlated with the treated/non‐treated (T/C) tumor size ratio (r=0.96, P<0.0001). In a second experiment, CDDP at 2.5 mg/kg b.w. and AGM‐1470 at 10 mg/kg b.w., these being the most effective doses, were given as in the first experiment, and animals were serially killed on days 14, 21, 28, 35 and 42. Tumors in rats treated with CDDP and AGM‐1470 showed 18.2% and 20.5% of the control telomerase activity on days 35 and 21, respectively, when tumor growth was inhibited. However, on day 42, the activities increased to 46.5% and 92.5%, this correlating with re‐growth (r=0.73, P<0.0001). These results suggest that decline of telomerase activity may be involved in tumor growth retardation induced by chemotherapeutic agents. This possibility clearly warrants further mechanistic studies.

Little finger replantation in a 20-month-old child: A case report

Abstract A case report of successful replantation of a completely amputated little finger in a 20-month-old child is presented.

Neural crest origin of clear cell sarcoma of tendons and aponeuroses: ultrastructural and enzyme cytochemical study of human and nude mouse-transplanted tumours

In order to clarify the histogenesis of clear cell sarcoma of tendons and aponeuroses (CCS), two cases of human and one nude mouse-transplanted CCS line were studied using an ultrastructural and enzyme cytochemical approach. Most of the tumour cells obtained from the primary and transplanted CCS demonstrated melanosomes in various stages of development within the cytoplasm, whereas no melanosomes could be identified in the metastatic CCS. However, cholinesterase and tyrosinase activities could be demonstrated not only in the melanotic primary and transplanted CCS but also in the amelanotic metastatic CCS. The results therefore support the hypothesis that CCS is a soft tissue tumour derived from the neural crest.