Koh Nakazawa

Hydronephrosis associated with retroperitoneal fibrosis and sclerosing pancreatitis

Sclerosing pancreatitis is associated with raised concentrations of IgG4. We treated 22 patients with sclerosing pancreatitis, and identified and followed-up three with concomitant hydronephrosis caused by ureteral mass, later diagnosed as retroperitoneal fibrosis. We histologically examined the ureteral and pancreatic lesions of these patients and noted abundant infiltration of IgG4-bearing plasma cells in both tissues. Treatment with corticosteroids lowered serum concentrations of IgG4. IgG4 might also have a pathological role in a systemic fibrosing process that includes pancreatic and retroperitoneal lesions.

An Autopsy Case of Crow-Fukase (POEMS) Syndrome with a High Level of IL-6 in the Ascites Special Reference to Glomerular Lesions

A 58‐year‐old man developed polyneuropathy, or‐ganomegaly, gynecomastia, skin pigmentation, and multiple myeloma (IgG‐Λ type). Although transient clinical improvement was obtained with prednisolone and cyclophosphamide, his general condition deteriorated progressively, and he died 3 years after onset. Lymph nodes showed angiofollicular hyperplasia, and the sural nerve revealed segmental demyellnation. At autopsy, the glomeruli were enlarged and showed mild mesangial cell proliferation with segmental mesangiolysis and proliferation of endothellal cells forming abnormal vessels. The massive volume of ascites contained a high level of inter‐leukin‐6 (IL‐6). IL‐6, a multifunctlonal cytokine, may be the pathogenic factor which induces the various clinical symptoms and pathological features of Crow‐Fukase (POEMS) syndrome. Acta Pathol Jpn 42: 651–656,1992.

Ultrastructural localization of calmodulin in gerbil cochlea by immunogold electron microscopy.

Localization of calmodulin, a calcium binding protein, was identified in adult gerbil cochleas using paraffin section immunohistochemistry and immunogold electron microscopy with monoclonal antibody against bovine calmodulin. Immunoreactive calmodulin was abundant in inner hair cells (IHCs), outer hair cells (OHCs) and Boettcher cells of the cochleas. Other cell types containing calmodulin were marginal cells and basal cells of the stria vascularis, fibrocytes in the spiral ligament, spiral ganglion neurons and vascular smooth muscle cells. Immunogold labeling for calmodulin was observed in cuticular plate, stereocilia, and within cytoplasm of IHCs and OHCs. In OHCs the labeling was mostly observed in the region underlying lateral wall corresponding to subsurface cisterna. In IHCs the staining was diffuse in the cytoplasm and denser than that in OHCs. Boettcher cells showed dense staining along the microvillous projections facing to the intercellular spaces between Boettcher cells and Claudius cells and between the neighboring Boettcher cells. These distributions of calmodulin in the hair cells consist with the assumption that IHCs act as a true neurotransducer and OHCs as an active bi-directional mechanotransducer. The rich presence of calmodulin in Boettcher cells suggests that the cells may involve in mediating Ca(2+) regulation and play a distinctive active role in ion transport.

Three-dimensional ultrastructure of glomerular injury in serum sickness nephritis using the quick-freezing and deep-etching method

The three-dimensional ultrastructure of the glomerulus in serum sickness nephritis has been investigated by the quick-freezing and deep-etching method. Compact granular immune deposits were localized in filamentous networks in the lamina densa and mesangial matrices. These constitutional fibrils with diameters of 8–15 nm, were directly attached to the immune deposits. The filamentous networks became markedly loosened around the deposits. In podocytes, reticular microfilaments with positive decoration by myosin subfragment 1 (S1) were increased in flattened foot processes and directly attached to the cell membranes. Fine filaments with diameters of 4–7 nm were undecorated by S1 and connected with actin filaments as cross-bridges. Intermediate filaments were also increased in the cell bodies and primary processes of podocytes. Connecting fibrils in lamina rara externa were partially disrupted. The immune deposits were primarily detected in the networks of lamina densa and actually destroyed the size barrier composed of filamentous networks. Moreover, the mesangial deposits also disorganized mesangial networks to probably alter mesangial flow through the matrices. Increased actin filaments in foot processes seemingly reinforced the cell membranes and the connecting fibrils in lamina rara externa, which prevented the initial detachment of podocytes from the basement membrane.

Glomerular expression of α-smooth muscle actin reflects disease activity of IgA nephropathy

To elucidate the relationship between histological disease states and clinicopathological features in immunoglobulin A nephropathy (IgAN), 90 needle‐biopsy specimens diagnosed as IgAN were analyzed. The specimens were divided into four groups according to histological grade and stage index. Immunohistochemical features of α‐smooth muscle actin (α‐SMA), macrophages positive for myeloid/histiocyte antigen (MAC387), and expression of type I, III and IV collagens were all examined. Glomerular expression scores of α‐SMA and the degree of intraglomerular macrophage infiltration were highest in the active and non‐sclerotic groups. Type I and IV collagens were significantly more abundant in the sclerotic groups than in the active groups. Type III collagen was strongly expressed in both the active and sclerotic groups. Double immunolabeling of α‐SMA and intercellular adhesion molecule (ICAM)‐1 revealed that ICAM‐1 was expressed around the α‐SMA‐positive mesangial area. In multivariate analysis, the glomerular expression score of α‐SMA was mostly correlated with histological grading in the 10 clinicopathological parameters. Type IV collagen score was mostly correlated with histological staging. These results suggest that glomerular α‐SMA expression reflects the histological activity of IgAN. Immunohistological staining of α‐SMA is valuable to estimate the degree of disease activity in IgAN.

Loss of glomerular anionic sites and the development of albuminuria in rats with streptozotocin-induced diabetes.

Examination was made of changes in the anionic sites of the glomerular basement membrane (GBM) in rats with streptozotocin (STZ)-induced diabetes by the immersion method of polyethyleneimine (PEI). PEI particles in GBM of diabetic rats significantly decreased from the 1st through the 8th week. Urinary albumin excretion in diabetic rats significantly increased at the 2nd but not earlier week. Insulin treatment effectively prevented decrease in PEI particles in STZ-injected rats. In rats with STZ-induced diabetes, initial renal alteration was disturbance of the charge barrier, followed by the development of albuminuria. Continued deterioration of anionic sites and possibly additional disturbance of size barrier were considered responsible for the development of albuminuria. Insulin treatment appears to prevent the loss of anionic sites of GBM.

Preexisting Membranous Nephropathy in Allograft Kidney

A case of membranous nephropathy, preexisting in a donor kidney, will be reported. A 41-year-old man underwent a cadaver renal transplantation. An allograft biopsy specimen obtained during the operation showed spike formation on periodic acid-silver methenamine staining and deposition of IgG along the glomerular capillary loop on immunoperoxidase staining. Immunofluorescence staining for IgG remained in the specimens obtained on day 11 and after 4 weeks, but markedly decreased in the specimen obtained 7 weeks after transplantation. Electron-dense deposits also decreased in amount, but irregular thickening of the glomerular basement membrane with spikes, electron-lucent washout lesions, and small amounts of electron-dense deposits remained 20 months after the transplantation. These findings suggest that membranous nephropathy, as well as IgA nephritis and diabetic nephropathy, resolve after renal transplantation and that deposition of IgG markedly decreases within a few months after transplantation, but that complete histological restoration of the basement membrane needs at least a few years.

Immune deposits in the glomerular extracellular matrix detected by the quick-freezing and deep-etching method

Chronic serum sickness nephritis was induced in rats by sensitization with egg albumin. The glomeruli were then examined by the quick-freezing and deep-etching method with immunohistochemical identification of immune complex deposits on replica membranes. In control rats, the glomerular basement membrane showed a three-layered structure. The middle layer was composed of a compact meshwork of fine fibrils that was connected to adjacent endothelial and epithelial cells by perpendicular fibrils in the inner and outer layers. The mesangial matrix contained a similar but looser meshwork structure. In the nephritic rats, small granular deposits were observed within the meshwork, distorting its fine structure. Larger nodular aggregates extended continuously from the middle layer to the epithelial side. Disruption of the connecting fibrils overlying these larger aggregates was noted. The deposits were stained by antiegg albumin or antirat IgG antibodies. Gold particles immunostaining for the sensitizing antigen were also localized within the deposits. These findings suggest that the deposition of immune complexes occurs in the fibrillary meshworks of the mesangium and lamina densa in this experimental model, with the resultant distortion of their meshwork structures and the formation of nodular aggregates.

A Case of Granulocyte-Colony Stimulating Factor-Producing Hepatocellular Carcinoma Confirmed by Immunohistochemistry

Granulocyte-colony stimulating factor (G-CSF) is a naturally occurring glycoprotein that stimulates the proliferation and maturation of precursor cells in the bone marrow into fully differentiated neutrophils. Several reports of G-CSF-producing malignant tumors have been published, but scarcely any in the hepatobiliary system, such as in hepatocellular carcinoma (HCC). Here, we encountered a 69-yr-old man with a hepatic tumor who had received right hepatic resection. He showed leukocytosis of 25,450/µL along with elevated serum G-CSF. Histological examination of surgical samples demonstrated immunohistochemical staining for G-CSF, but not for G-CSF receptor. The patient survived without recurrence for four years, but ultimately passed away with multiple bone metastases. In light of the above, clinicians may consider G-CSF-producing HCC when encountering patients with leukocytosis and a hepatic tumor. More cases are needed to clarify the clinical picture of G-CSF-producing HCC.

Participation of endothelial cells and transformed mesangial cells in remodeling of glomerular capillary loops in Thy-1 nephritis

The relationship between mesangial cells (MC) and endothelial cells (EC) in the remodeling of glomerular capillary loops was investigated in a rat model of anti‐Thy‐1 antibody (Ab)‐induced glomerulonephritis. Immunohistochemical analysis showed that cells positive for α‐smooth muscle actin (α‐SMA) appeared in the mesangial stalks at day three, and had increased in number at day seven, after injection of Thy‐1 Ab. Double staining for α‐SMA and proliferating cell nuclear antigen (PCNA) showed that some MC expressing PCNA were negative for α‐SMA at day three, but by day seven almost all PCNA‐positive MC expressed α‐SMA. Western blotting for α‐SMA from isolated glomeruli was negative at day one after injection of Thy‐1 Ab, but positive at day seven. Type III collagen appeared at day seven, followed by an increase of EC in the capillary loops, as determined by double immunofluorescent staining for rat endothelial cell antigen‐1 (RECA‐1) and type III collagen. RECA‐1‐positive cells increased rapidly in number after day seven and eventually showed the same distribution pattern as that in control rats. Both type I and type III collagens were expressed in the mesangial and the ballooning area of the glomerulus at day seven. Electron microscopy revealed that immature MC and EC forming small capillary lumina appeared in the enlarged mesangial area at day seven. In accordance with the increase of capillaries and the enlargement of the lumina, the number of MC and the amount of mesangial matrix decreased gradually, and most of the glomeruli returned to a normal structure by week 4. These data show that type I and type III collagen produced by transformed MC may be of benefit to proliferation of EC and remodeling of the capillary in Thy‐1‐induced nephritis.