Tissue and cellular distribution of alpha-L-iduronidase in the pig.

We investigated the alpha-L-iduronidase activity of various pig tissues. Furthermore, we examined the tissues using antibody, enzyme immunoassay (EIA), and immunohistochemical methods. The amounts of enzyme measured by the EIA method in the various tissues were proportional to their enzyme activities and also to their immunohistochemical characteristics. The tissues could thus be classified into three groups: a high enzyme activity group composed of the liver, kidney, and spleen; a moderate activity group comprising the lung, lymph nodes, stomach, ileum, colon, and pancreas; and a low activity group consisting of the heart, diaphragm, iliopsoas muscle, cerebrum, cerebellum, and skin. The molecular weight of the enzyme in each tissue did not reveal any heterogeneity, having two components of 70 KD and 62 KD by Western blot analysis. Immunohistochemically, alpha-L-iduronidase was strongly detected in the lysosomal membranes of cells of the mononuclear phagocyte system, epithelial cells of the proximal tubules in the kidney, and some blastic cells, whereas hepatocytes revealed weak positive reactions. The tissue and cellular distribution of the enzyme appeared to have a close relation to tissues that manifest or are affected by alpha-L-iduronidase deficiency.

A new devised skirted elephant trunk technique.

A simple and effective new elephant trunk technique was devised and applied to two patients with a successful result. In advance before the operation, an arch graft with a skirted elephant trunk was made. This was done by inserting a smaller, 22 mm diameter sized graft into the arch graft at the distal end and suturing it so as to leave a skirt extending over the smaller graft. This configuration facilitates the distal anastomosis and effectively shortens anastomotic time.

A ruptured syphilitic descending thoracic aortic aneurysm. The characteristic findings on computed tomography for the etiological diagnosis of aneurysm.

We report the case of a 72-year-old man with a ruptured syphilitic descending thoracic aneurysm who underwent an emergency operation and successful graft replacement. Preoperative physical examination showed a pulsative mass on the left back. Preoperative computed tomography showed bone destruction in the TH6 to TH10 thoracic vertebrae and ribs and penetration (or rupture) of the aneurysm into the subcutaneous tissue. During the period of preoperative evaluations, free wall rupture of the aneurysm occurred and emergency operation for graft replacement was performed. The microscopical examination of the aneurysmal wall revealed the syphilitic changes. In literature, the vertebral destruction by atherosclerotic aneurysm is usually located at the TH12 to L3 of vertebral bodies. From the findings of this patient and a study of existing literature, we concluded that the finding of vertebral bone beyond TH12 to L3 region on CT examination of the aneurysm could be a etiological characteristic finding for syphilitic aortic aneurysm.

[Thoracoscopic surgery for diagnosis of interstitial pneumonia special reference to medicoeconomical effect].

We compared thoracoscopic surgery (TS) and open thoracotomy for the diagnosis of interstitial pneumonia. Intraoperative blood loss and duration of postoperative chest drainage were significantly less with TS than with thoracotomy. The length of postoperative hospital stay and social insurance costs with TS was significantly less than with thoracotomy. These results show that TS for the diagnosis of interstitial pneumonia is superior to open thoracotomy in terms of surgical stress and cost.

[Glycogenosis type II; acid alpha-glucosidase deficiency].

Knowledge of the general enzymology of lysosomal acid alpha-glucosidase (acid maltase) including that of the intracellular processing mechanism and of the cDNA sequence has advanced recently. Genetic heterogeneity of the mRNA of acid alpha-glucosidase deficient patients has also been reported. Referring to these recent advances, and based on our previously reported results, the pathologic aspects of the disease are reviewed. Heterogeneity of the enzyme molecules themselves and their characteristics in various tissues are also demonstrated.

Purification, characterization and subcellular localization of pig liver alpha-L-iduronidase.

alpha-L-Iduronidase was purified about 100,000-fold from pig liver by employing column chromatography on cellulose phosphate (P11), concanavalin A-Sepharose 4B, heparin-Sepharose 4B, Toyopearl HW-55, Sephadex G-100 and chelating Sepharose 6B charged with cupric ions. The molecular mass of the purified enzyme was estimated to be 70 kDa by Sephadex G-100 column chromatography. The purified enzyme gave a single band on disc polyacrylamide gel electrophoresis without using sodium dodecyl sulfate. However, two separate components of 70 kDa and 62 kDa appeared when it was analyzed by SDS/polyacrylamide gel electrophoresis. These 70-kDa and 62-kDa components were confirmed as alpha-L-iduronidase immunochemically. The isoelectric points of these enzymes were both 9.1 as measured by isoelectric focusing in a polyacrylamide gel containing ampholine and sucrose. The optimal pH and Km values were 3.0-3.5 and 65 microM 4-methylumbelliferyl-alpha-L-iduronide, respectively. The purified enzyme was stable in the pH range 3.5-6.0 under conditions with or without 0.5 M NaCl. However, in the presence of 0.5 M NaCl, it was unstable at pH 3.0. Moreover, it was conversely stabilized at pH 7.0 in the presence of 0.5 M NaCl. Immunohistochemically, the enzyme was found in the Kupffer cells and was abundant on their lysosomal membranes. In liver cells, however, the immunohistochemical reaction was weak.

Media conditioned by smooth muscle cells cultured in a variety of hypoxic environments stimulates in vitro angiogenesis. A relationship to transforming growth factor-beta 1.

Conditioned media (CM) harvested from bovine smooth muscle cells (SMCs) of aortic media cultured under hypoxic conditions remarkably enhanced angiogenesis in vitro, that is, the tube formation of bovine capillary endothelial cells (BCEs) cultured on type I collagen gels. The extent of in vitro angiogenesis was assessed by the total length of tube structures formed by BCEs per area measured quantitatively with an image analyzer. The tube formation in CM obtained from the cultivation of SMCs at 1% O2 for 24 hours was enhanced by about 1.5 times and 3.4 times as compared with those at 5% O2 and 20% O2, respectively. This tube-forming activity was abrogated by the pretreatment of CM with anti-transforming growth factor (TGF)-beta 1 IgG, but not by anti-basic fibroblast growth factor IgG. The SMC-CM obtained from hypoxic cultivation (1% O2 for 24 hours) inhibited [3H] thymidine incorporation by BCEs, SMCs, and fibroblasts more than about 20% of control. Anti-TGF-beta 1 IgG thus significantly reduced the inhibitory effect of hypoxic SMC-CM on DNA synthesis of these cells. These results suggest that SMCs in a hypoxic state release active in vitro angiogenic factors into CM, and active TGF-beta 1 is closely related to the in vitro angiogenic enhancement of media conditioned by SMCs cultured in a hypoxic state.

Lectin reactivity of alpha-fetoprotein in a case of renal cell carcinoma.

The increased serum level of alpha-fetoprotein (AFP) in a case of renal cell carcinoma, a rare condition of AFP production by mesoderm-derived cells, was evaluated for its lectin reactivity by affinity electrophoresis, followed by the antibody-affinity transfer to nitrocellulose membranes for visualization of separated AFP bands. The AFP of this case was characterized by relative increases of concanavalin A-nonreactive AFP-C1 (60.4%), erythroagglutinating phytohemagglutinin-reactive AFP-P4 (37.8%) and AFP-P5 (46.3%) and Allomyrina dichotoma lectin-nonreactive AFP-A1s (66.7%), and by the total absence of lentil lectin-reactive components, AFP-L2 and AFP-L3. Thus, the lectin-reactive pattern of AFP markedly deviated not only from that of cord serum, but also from those of other malignancies and of fetal kidney cells in culture.

Acute ascending necrotizing myelitis in Okinawa caused by herpes simplex virus type 2.

A case of rapidly progressing ascending myelitis was necropsied. Necrosis was present throughout the whole length of the spinal cord and involved both the grey and white matter randomly. The perivascular lymphocytic infiltration in the spinal cord in the present case was more pronounced than that in the previously reported two cases of necrotizing myelopathy associated with malignancy. Using immunoperoxidase staining the presence of herpes simplex virus type 2 (HSV 2) antigen was demonstrated. Electron microscopic examinations revealed large numbers of HSV particles in the spinal cord. HSV 2 may be a common aetiological agent of necrotizing myelopathy and myelitis in Okinawa, an HSV 2 endemic area. In the present case, the necrosis was mainly found in the spinal cord but was also observed, to a very limited extent, in the brain.

Two cases of necrotizing myelopathy associated with malignancy caused by herpes simplex virus type 2.

Two cases of necrotizing myelopathy were autopsied; one was complicated with lung carcinoma and the other with chronic type adult T cell leukemia (ATL leukemia). To our knowledge, they were the first cases of their type in Japan. In both cases, necrosis of the spinal cord was observed in the gray and white matter along most of its extent. Marked changes were found in the lumbar segment. The patients were not treated with intravenous cancer chemotherapy or irradiation. Immunohistochemical and electron microscopic examination revealed an extremely strong infection of herpes simplex virus (HSV) type 2. However, HSV type 1 and cytomegalovirus antigens were not detected.