K. Iwasaki

LEU-7 IMMUNOREACTIVITY IN HUMAN AND RAT EMBRYONIC HEARTS, WITH SPECIAL REFERENCE TO THE DEVELOPMENT OF THE CONDUCTION TISSUE

SummaryThe distribution pattern of Leu-7 (HNK-1) in developing human embryonic hearts and rat hearts was studied by immunohistochemistry. Human and rat embryos at Streeters stages XIII ∼ XX and fetus stage I were used. Leu-7, which is absent in the newborn rat heart, is expressed transiently in the embryo and fetus I stages. The earliest embryonic heart shows two incomplete circular structures with immunoreactivity in the myocardium along the primitive atrioventricular cushion and bulboventricular canal. These two structures become localized topographically in the definitive atrioventricular node and atrioventricular bundle after rearrangement and partial disappearance during embryonic development. At Streeters stages XVIII ∼ XX, Leu-7 immunoreactivity appears to localize topographically in almost all the pathways of the conduction system, although some discontinuities are observed in the atrioventricular junction and atrial internodal tracts. Thereafter, immunoreactivity decreases gradually and differentially by site and stage. The precise nature of Leu-7 immunoreactive cells, that is, whether or not they are neurogenic or myogenic, is not revealed by this study. The present observations are discussed in connection with the hypothesis that specialized ring tissue is the primordium of the conduction system.

Femoral head necrosis and osteopenia in stroke-prone spontaneously hypertensive rats (SHRSPs)

Necrosis of the femoral head and osteopenia were examined histopathologically in stroke-prone spontaneously hypertensive rats (SHRSPs) aged 6 to 36 weeks and compared with that of spontaneously hypertensive rats (SHRs) and Wistar-Kyoto rats (WKYs). Avascular necrosis of the femoral head was frequently observed, mainly in the young SHRSPs and SHRs (about 8 to 15 weeks of age). SHRSPs had the highest incidence of femoral head necrosis among the three strains. This necrotic change in the femoral head was considered to be secondary ischemia induced by angiospasm or arteriosclerosis, similar to the disorders observed in the brain, kidney, and heart in SHRSPs. However, the complication occurred in spite of treatment with antihypertensive agents (ACE inhibitor: enalapril, spirapril) even though other ischemic disorders such as brain hemorrhage and renal infarction were prevented, indicating that the femoral head necrosis in SHRSPs was not due to hypertensive complications induced by angiospasm or arteriosclerosis. Bone mineral density (BMD) of the femoral bone was significantly lower in SHRSPs, and the femoral heads in this strain were the most easily deformed by loads applied during compression tests. Histopathologically, the infarctions were encountered on the lateral side of the epiphysis, but no thrombi were observed. The lateral side of the epiphysis is the anatomic site where the weight load is greatest and the site where the nutritive artery enters. Our results strongly suggest that the coexistence of vulnerable bone matrix and physical weight load to the nutritive artery plays a crucial role in the occurrence of femoral head necrosis in SHRSPs, whether based on generalized or localized osteopenia.

HNK-1 expression pattern in normal and bis-diamine induced malformed developing rat heart: three dimensional reconstruction analysis using computer graphics

SummaryThe spatiotemporal distribution of the immunoreactivity of monoclonal antibody HNK-1 was investigated immunohistochemically in normal and bis-diamine-induced malformed rat embryonic hearts using three-dimensional reconstruction with computer graphics. First recognized in the primitive heart 11.5 days after conception, HNK-1 immunoreactivity was distributed in the atrio-ventricular and bulbo-ventricular junctional areas with incomplete ring-like appearance in the early embryonic stages. In the late embryonic stages the immunoreactive sites were rearranged and localized in the sites topographically corresponding to almost the entire pathway of the conduction system, including the three major internodal tracts connecting the right sinoatrial node and atrioventricular node. Immunoreactivity gradually decreased after the completion of the conduction system, and only a faint reactivity in the atrio-ventricular node region remained in the new-born heart. These results indicate that HNK-1 is expressed temporarily in the pathways corresponding to the conduction system during the development of the heart. In bis-(dichloro-acethyl)-octamethylen-diamine (bis-diamine)-induced malformed hearts, localization of HNK-1 immunoreactivity was not remarkably altered in the early embryonic heart. In the late embryo, immunoreactive sites in the sino-atrial node region and atrio-ventricular node region deviated dorsocaudally with the poorly developed internodal tracts, and abnormal distribution was observed in the bilateral atria. We consider that these abnormalities may occur in conjunction with abnormal morphological development such as insufficient absorption of the sinus venosus.

Distribution of acetylcholinesterase activity in the rat embryonic heart with reference to HNK-1 immunoreactivity in the conduction tissue

Acetylcholinesterase (AChE) activity was topographically investigated in the presumptive cardiac conduction tissue regions visualized by HNK-1 immunoreactivity in rat embryos, and AChE-positive cells were examined with the electron microscope. On embryonic day (ED) 14.5, when HNK-1 was most intensely visualized, AChE activity could not be detected enzyme-histochemically in the conduction tissue regions, except in the ventricular trabeculae and part of the AV node. On ED 16.5, however, the AChE activity was clearly demonstrated in some parts of the developing conduction tissue. One exception was the AV node region, where an AChE-positive area was in close proximity to an area showing HNK-1 immunoreactivity but did not overlap. Furthermore, AChE activity was demonstrated predominantly in the ventricular trabeculae, including cardiac myocytes, but was rather weak in the atrium. With the electron microscope, AChE reaction products were observed predominantly intracellulary in both developing conduction tissue cells and developing ordinary myocytes, and no reactivity was found in neuronal components. From ED 18.5 until birth, both AChE activity and HNK-1 immunoreactivity faded away in the conduction tissue. Thus, transient AChE activity in the embryonic heart seems to be different from the developing adult form and may be related to a morphogenetic function in embryonic tissues, as proposed by other authors.

Vascular occlusion and cartilage disorders in osteonecrosis of the femoral head in rats

Summary. The aim of our study was to clarify the role of lateral epiphyseal vessels in the development of femoral head osteonecrosis in rats. We did this by performing histological and immunological studies on rats at the age that they are most prone to spontaneous osteonecrosis. We observed that obstruction of the lateral epiphyseal vessels occurred at the site of cartilage penetration. This obstruction preceded the development of osteonecrosis in adjacent areas of the femoral head. We propose that a similar mechanism may be responsible, in part, for the development of Perthes’ disease in man.

Effects of radiation on chondrocytes in culture

Using an in vitro model, we studied the direct effects of radiation on both DNA and glycosaminoglycan (GAG) synthesis of chondrocytes. Chondrocytes from articular cartilage of 21-day-old rabbits were cultured in monolayer or in pellets with matrix as a three-dimensional tube culture. These cells were exposed to a single dose of X-rays at either 2 or 10 Gy. Following irradiation at 2 Gy, DNA synthesis in the chondrocytes was temporarily suppressed but rapidly returned to the control level, while at 10 Gy, DNA synthesis was markedly suppressed and there was no increase in cell number. On the other hand, GAG synthesis was not affected by a single dose of X-rays at either 2 or 10 Gy, nor did GAG content decrease in the three-dimensional tube cultures. These results show that radiation affects the proliferation and differentiation of chondrocytes differently, and that the synthesis of the components of cartilaginous matrix such as GAG is relatively radioresistant in contrast to DNA synthesis.

An electron microscopic examination of age-related changes in the rat liver. The influence of diet.

The influence of age and diet on the ultrastructure of hepatocytes is reported. The following dietary manipulations were investigated: Group 1, fed ad libitum a diet containing 21% protein; Group 2, fed a similar diet but restricted to 60% of the intake of Group 1 from 6 weeks of age onwards; Group 3, restricted from 6 weeks to 6 months of age and thereafter fed ad libitum; Group 4, restriction started at 6 months of age; Group 5, fed ad libitum a diet containing 12.6% protein. In all groups the size of hepatocytes was found not to increase during adult life. The size of hepatocytes in Groups 2 and 4 was the same as or larger than that of the other groups; thus food restriction resulted in a decreased number of hepatocytes. Changes in the structure of some organelles and the accumulation of lipofuscin granules occurred with advancing age and the extent of these age‐related changes was less in Groups 2 and 4 than in the other groups. These morphologic findings in conjunction with our previously reported metabolic findings provide a new view of the action of food restriction on the aging process. ACTA PATHOL JPN 38: 1119∼1130, 1988.

ENDOSCOPIC SUBMUCOSAL DISSECTION OF A MINUTE INTRAMUCOSAL ADENOCARCINOMA IN BARRETT'S ESOPHAGUS

A 73‐year‐old man with short segmental Barretts esophagus underwent esophagoscopy, and a slightly depressed, discolored lesion was found on the anterior wall of the lower esophagus. Under a provisional diagnosis of differentiated adenocarcinoma without local lymph node metastasis, endoscopic submucosal dissection (ESD) was carried out. En bloc resection with tumor‐free lateral/basal margins was accomplished without complication. The resected area was 12 × 15 mm in size, whereas the neoplastic lesion was 4 × 4 mm. Histopathological examination confirmed intramucosal well‐differentiated tubular adenocarcinoma without angiolymphatic invasion adjacent to the muscularis mucosae. Repeated esophagoscopy 6 months after ESD showed neither locally recurrent nor metachronous lesions. Considering that Barretts esophagus is a precancerous condition, one may recommend eradication of both the neoplastic and non‐neoplastic lesion with using ESD.

A Sequential Ultrastructural and Histoautoradiographic Study of Early Neoplastic Lesions in Ethylnitrosouyea‐induced Rat Glioma

A sequential study of the early stage of development of ethylnitrosourea (ENU) induced glioma in the rat was performed by electron microscopy and [3H]thymidine histoautoradiography. Hyperplasia, the earliest neoplastic change that was detectable morphologically, consisted of a few or several immature oligodendroglia like cells which were connected with one another or with preexisting neural tissue by junctional apparatus, and showed no reactive changes in the astrocytes or microglia. The labelling index of hyperplastic cells was 2.6%. Foci of early neoplastic proliferation (ENP) showed mild destructive changes in the neighboring neural tissue, and their major constituent cells had characteristics of immature oligodendroglias. The labelling index of cells showing ENP was 3.3%. The intercellular spaces exhibited slight enlargement with accumulation of extracellular matrix and a decrease in the number of junctional apparatus on the neoplastic cells. Microtumors showed apparent destruction of the preexisting neural tissue to form a tumor mass with an increase in the extracellular matrix. Constituent cells of the microtumors were similar to those of the ENP, although reactive astrocytes and microglias occurred more frequently. The labelling index was 9.6% in the central area of microtumors and 5.3% in the peripheral area. These findings suggest that in the initial or very early stages of glial cell neoplastic proliferation, it is necessary for the neoplastic cells to maintain contact with the neurons for metabolic purposes, and that after losing contact, these cells can proliferate autonomously with the accumulation of extracellular matrix.

A case of angiocentric immunoproliferative lesions (angiocentric lymphoma) associated with human T-cell lymphotropic virus type 1

We report a case of angiocentric immunoproliferative lesions (AILs; angiocentric lymphoma) in the subcutaneous tissue associated with human T-cell lymphotropic virus type 1 (HTLV-1). The patient showed an indolent cutaneous manifestation for 9 years until death. A skin biopsy revealed an angiocentric infiltration of atypical T lymphocytes and a marked coagulative necrosis, which are histologic hallmarks of AILs. Human T-cell lymphotropic virus type 1 antibody was detected by the indirect immunofluorescence test. The proviral integration of HTLV-1 also was confirmed in a DNA extract from paraffin sections using the polymerase chain reaction. The Epstein-Barr virus genome, which has been reported to be associated with AILs, could not be detected by polymerase chain reaction. The present case indicated that HTLV-1 also should be considered one of the pathogenetic factors of AILs.