Urinary UK, t-PA and urinary trypsin inhibitor in health and glomerular diseases

The concentrations of two different plasminogen activators(PAs), urokinase (UK), tissue-type plasminogen activator (t-PA) and urinary trypsin inhibitor (UTI) were determined in the urine and blood from 48 normal subjects and 92 patients with glomerulonephritis using highly sensitive enzyme immunoassay (EIA). The values of UK clearance were approximately 1.5-fold larger than those of creatinine clearance and at least 60.8% of UK was reabsorbed in the renal tubules, which suggest that one of major secretion site of UK is located in the outer region of the glomerular basement membrane (GBM), that is glomerular epithelium. Decreased urinary excretion of UK was observed in the glomerular disease depending on their severity and correlated with the increasing degree of FDP D-dimer excretion. On the other hand, the values of t-PA clearance were quite smaller than those of creatinine clearance, which suggest that urinary t-PA originated from the blood circulation or the inner side of the GBM (possibly glomerular endothelium) and filtrated from the GBM. Like UK, urinary t-PA also decreased in glomerular diseases. UTI which is highly anionic and has a comparable size with albumin was excreted increasingly in glomerulo-nephritis due to loss of the anionic charge barrier of the GBM. No significant correlations were noted between UTI excretion and UK or t-PA excretion.     

ENDOTOXEMIA-INDUCED DIFFUSE MYELITIS AND EXTENSIVE PATCHY NECROSIS OF THE LIVER

An autopsy case with endotoxemia induced diffuse myelitis and extensive, grossly patchy necrosis of the liver occurring in a 70-year-old female was examined histopa-thologically and electron microscopically. Leucopenia with prominent leukemoid reaction (myeloblasts 20%) preceded the terminal fulminant hepatitis by two weeks. Soon after the terminal event, bacteremia and endotoxemia were detected and negativity for HB antigen was proved. Diffuse myelitis was characterized by devastation of hyperplastic bone marrow structure mottled with destructed sinus architecture and scattered exudative necrosis, resulting in the loss of mature granulocytes and erythro-poiesis. Regenerative clusters of myeloblasts and prominent increase of megakaryocytes were observed. Electron microscopically, the bone marrow contained fibrin and platelets within the exudate of the marrow stroma. Extensive, grossly patchy necrosis of the liver microscopically consisted of well demarcated coagulation necrosis of hepatic parenchyma with scattered fibrin thrombi in the sinusoids at the boundary. There were no definite thrombi but occasional fibrin accumulation in the small blood vessels of the liver. Both extensive diffuse myelitis and extensive, patchy necrosis of the liver seemed to be quite rare in incidence. The pathogenesis of these combined lesions was discussed in relation with endotoxemia.     

The effect of bleomycin administered in combination with hyperthermia on a C3H mouse fibrosarcoma

The effect of bleomycin (BLM) on a murine fibrosarcoma, FSa-II, was investigated at elevated temperatures. Animals were C3Hf/Sed mice derived from our defined flora mouse colony. Single-cell suspensions of the tumour were transplanted into the murine foot. Tumours with an average diameter of 4 mm were treated in a water bath where a desired temperature was maintained. Some animal groups received an intraperitoneal injection of glucose 60 min before hyperthermia to reduce tumour tissue pH. Hyperthermia was given for 60 min at 41.5 degrees C, 30 min at 43.5 degrees C, or 60 min at 43.5 degrees C, and tumour growth (TG) time to reach 1000 mm3 was studied. BLM treatment at 41.5 degrees C for 60 min prolonged the TG time compared to BLM treatment at a room temperature. The identical prolongation was observed following BLM treatment given at 43.5 degrees C for 30 min, although the growth prolongation was greater following BLM treatment given at 43.5 degrees C for 60 min than the other two treatments. This greater prolongation is probably attributable to an additive effect; namely hyperthermia of 43.5 degrees C for 60 min induces substantial lethal damage by itself while the other two heat treatments do not induce significant damage. It is interesting to note that this kind of enhancement was observed at a low temperature of 41.5 degrees C. A glucose injection before BLM and hyperthermia treatments further enhanced tumour response. These observations agree with our previous observations following in vitro treatments of the same tumour cells, and indicate that BLM is a good drug to be combined with hyperthermia.     

Effect of thermochemotherapy (combined cyclophosphamide and hyperthermia) given at various temperatures with or without glucose administration on a murine fibrosarcoma

The effect of combined cyclophosphamide (CY) and heat treatments on a murine tumor was studied at various temperatures. FSa-II tumors, the early generation isotransplants of a spontaneous fibrosarcoma in a C3Hf/Sed mouse, were used. A single cell suspension was transplanted into the animal foot. Hyperthermia was given by immersing animal feet into a water bath maintained at a desired temperature +/- 0.1 degrees C. An average diameter of the tumor at the time of treatment was 4 mm. The tumor growth time, the time required for one-half of the treated tumors to reach 1000 mm3, was the end point. Hyperthermia enhanced the effect of CY at test temperatures ranging from 40.5 degrees - 44.5 degrees C. The enhancement was independent of the temperature when CY was administered 30 min before the beginning of hyperthermia. However, the enhancement was most substantial at temperatures of 40.5 degrees -42.5 degrees C when CY was administered immediately before hyperthermia. The most effective timing of the CY administration was immediately before hyperthermia. The glucose administered 60 min before hyperthermia enhanced the effect of combined CY and hyperthermia when CY was given 30 min before heating. This enhancement was lost when CY was given immediately before hyperthermia. The CY dose response curves at elevated temperatures were downward concave, which may indicate the presence of a CY- and heat-resistant cell population in the tumor. Implications of these observations in clinical hyperthermia were discussed.     

Monoamine oxidase and acetylcholinesterase in the neurohypophysis of the hagfish, Eptatretus burgeri

The peripheral region of both dorsal and ventral walls of the neurohypophyseal sac of the hagfish, Eptatretus burgeri, showed strong monoamine oxidase (MAO) activity. Acetylcholinesterase (AChE) activity was detected only in the dorsal wall anterior to the infundibular stalk and in the ventral wall, except for the ependymal cells. AChE was not detected in the dorsal wall posterior to the stalk, where aldehyde-fuchsin-positive material is deposited. From these distribution patterns of MAO and AChE the identity of the structural specializations of the neurohypophysis are discussed as compared with distributions of MAO and AChE in the median eminence and the pars nervosa of higher vertebrates.