Case report. Cryptococcal cellulitis showing necrotizing vasculitis.

A 65-year-old woman with refractory anaemia who had been treated with systemic corticosteroids for several months developed cryptococcal cellulitis of the right cubital fossa. She was treated empirically for a presumed bacterial cellulitis with little response. Histological examination of debrided tissue revealed Cryptococcus as the causative organism. The tissue reaction involved suppurative inflammation with abscess formation in the dermis and subcutaneous adipose tissue. Necrotizing vasculitis, which has rarely been described in cryptococcosis, was seen in this case. Although the cellulitis was cured by local treatment in this patient, most previous reports recommended systemic antifungal therapy to treat cryptococcal cellulitis.

Toxicity of cisplatin to the central nervous system of male rabbits.

The cytotoxic effects of platinum (Pt) were studied by intraparenchymal injection of 1 mg of cisplatin (CDDP) in male rabbits. Time-serial plasma Pt levels were used as CDDP clearance indices in brain and kidney tissues. The tissue samples were also examined histologically. Changes in the blood-brain barrier (BBB) were evaluated by horseradish peroxidase (HRP) extravasation. In the brain infusion group, Pt was detected in the plasma 30 min after the start of infusion. In the kidney, Pt was detected after 10 min of CDDP injection. The maximum plasma concentration of Pt in the brain group showed diffuse edema, neuronal necrosis, karyolysis, and HRP extravasation around the injection site. In contrast, the histological damage to kidneys was minimal. The results presented here show that direct infusion of CDDP caused the most extensive cytotoxicity in the brain. The low clearance rate of CDDP from the brain and BBB disruption may explain this behavior.

Biological functions of extravasated serum IgG in rat brain.

During blood-brain barrier opening serum IgG could be extravasated. The function of intraparenchymal IgG, however, is unknown. Its biological effects in the acute phase were currently investigated. From rat autoserum IgG was purified and injected into the cortex. Similarly, IgG-Fab fragment was prepared and administered likewise. As for the control group, only vehicle was injected. Animals were sacrificed on days 1, 2 and 4 after the infusion and were histologically evaluated. On days 1 and 2, the infusion of IgG caused significant intraparenchymal infiltration of neutrophils which expressed LFA-1-alpha. It also induced CR3 up-regulation in microglia and endothelial ICAM-1 expression. On day 4, these findings had disappeared. HE stained brain sections and the TUNEL method did not reveal significant nerve cell death in IgG injected animals during the experiment as compared to the controls. IgG-Fab did not cause significant changes either. Extravasated IgG has been viewed to have biochemical functions. Its Fc fragment seemed to cause microglial and endothelial activation, followed by leukocytic infiltration. This sequence itself was not neurotoxic. Therefore, it is suggested that extravasated IgG is one of the inducers that modulate cellular responses in the acute phase of brain damage.

[Effects of allopurinol for oxidative injury of cisplatin-induced nephrotoxicity in mice].

The effects of allopurinol (Allop) on the lipid peroxidation in the nephrotoxicity of an antitumor drug, cisplatin (CDDP) were studied in mice. CDDP was administered intraperitoneally to two groups (CDDP + Allop group and CDDP + CMC-Na group) at single doses of 10 mg/kg, and mice were sacrificed 3 days after CDDP administration. The body weights of the CDDP-administered group gradually decreased to approximately 78% of the values of the control group (saline + Allop group and saline + CMC-Na group) within 3 days. Plasma urea nitrogen and creatinine, especially in the CDDP + Allop group, increased after 3 days. Lipid peroxides in the blood and kidney were monitored by measuring the production of malondialdehyde (MDA), which increased in the CDDP + CMC-Na group. On the other hand, MDA levels in the CDDP + Allop group increased in the kidney but remained unchanged in the blood. Changes were observed in tissue glutathione (reduced form, GSH; oxidized form, GSSG) levels in the CDDP + Allop group but not in the CDDP + CMC-Na group. Histomorphological examination demonstrated the degeneration of the proximal tubuli in the CDDP-administered groups. Especially in the CDDP + Allop group, the increase of mesangium cells in the glomeruli was observed. From these results, it was suggested that Allop was not able to inhibit CDDP-induced lipid peroxidation in the kidney, and the kidney function became more severely impaired by the administration of Allop.

Comparison of commercial isotonic fluids intravenously administered to rehydrate fasted bullocks.

Holstein bullocks were used in this study to compare the effectiveness of five commercial parenteral fluids (saline IS, Hartmann's IH, 5%-glucose 5G, Ringers IR, and 1/2 Ringer's and 2.5% glucose combination solutions RG) in correcting the disturbances associated with dehydration induced by fasting for 48 hr. These five commercial fluids (30 ml/kg) were given to bullocks with dehydration induced by fasting for 48 hr. Arterial and venous blood samples were taken before fasting, and at 0, 15, 30, 45, 60, 90, 120, 240, 360 min, and 24 hr after initiation of fluid administration. Fasting for 48 hr induced significant reductions in body weight and relative plasma volume (rPV), of approximately 7.72 and 21.93%, respectively. During the administration period, rPV showed a progressive increase from approximately 88.1% after fasting to 113.0% with no significantly differences between groups. A rapid decrease of rPV when fluid administration has been finished was observed in the 5G and RG groups. The results of the fluid administration trial showed that the 1/2 Ringer's and 2.5% glucose combination solution inhibited the acidification of the blood, produced no change in the electrolyte balance of serum, and induced a proper reabsorption rate of glucose in the renals, and was therefore considered the best choice for the rehydration of adult cattle which have had no appetite for over 2 days.

Tissue platinum after clinical treatment with cisplatin or carboplatin in tumor-bearing patients.

Tissue platinum (Pt) levels were measured in tumor-bearing patients treated with either cisplatin or carboplatin. Cisplatin was given by intra-arterial, intraperitoneal, and intravenous (iv) administrations. After death, vertebrae and intervertebral disks were removed from eight human subjects, and livers and kidneys were removed from the half of them. When cisplatin was administered intraperitoneally, Pt of the liver was higher than that of the kidney, and a high content of Pt was detected in the vertebra by comparing with the other administration methods. At the intra-arterial administration of cisplatin, Pt was mainly accumulated in the kidney. At the iv administration of cisplatin, a high level of Pt was found in the vertebra and intervertebral disk, especially at the highest value at 10.31 micrograms/g in the intervertebral disk of one case, whereas a low level of Pt was detected in the liver. On the contrary, it was found that the iv administration of carboplatin did not result in high accumulations of Pt in the liver, kidney, intervertebral disk, and vertebra. Therefore, Pt is accumulated in different organs, depending on the way cisplatin is administered, but Pt is accumulated least in them by the administration of carboplatin.

Novel animal model of lymph node metastasis by intrauterine inoculation of the actively metastatic subline PL3 separated from rat Walker 256 tumor cells.

To investigate the cellular mechanism of lymph node metastasis by tumor cells through the lymphatic vessels in the uterine corpus, we selected an active metastatic subline (PL3) from rat Walker 256 tumor cells and used it to develop a novel experimental model of lymph node metastasis induced by intrauterine inoculation of the tumor cells. Light- and electron-microscopic examinations revealed that the inoculated PL3 cells could actively infiltrate the endometrium from the uterine cavity and form a primary lesion in the uterine corpus. A few PL3 cells in the myometrium were found in the lumen of the peripheral lymphatic vessels on day 7 after inoculation. The regional lymph nodes around the uterus were then invaded by the migrated PL3 cells, and finally (after 3 weeks), most of the parenchyma of the nodes was replaced by metastasized tumor cells. By flow-cytometric analysis, the metastatic PL3 cells expressed CD44, like Walker 256 cells, but lacked integrin alphaL- and alpha4-chains. However, expression of ICAM-1 was considerably down-regulated in the PL3 cells compared to the parent cells. More aggressive invasion was shown by the PL3 cells compared to the parent cells in the in vitro invasion assay. These findings suggest that this experimental model and the separated PL3 cells are suitable for thorough investigations of the unidentified metastatic process and the related cellular behavior involved in the onset of lymphatic invasion by the primary lesion. Furthermore, our model more closely reproduces the clinical conditions related to lymph node metastasis of malignant carcinomas through the lymphatic vessels than does any previously reported animal model.

Neurotoxicity of serum components, comparison between CA1 and striatum.

In vasogenic brain edema, the neurotoxicity of extravasated serum components may contribute to neuronal damage. In the hippocampal CA1 sector and striatum, the neurotoxicity of serum was investigated. Rat serum was prepared as follows: Serum-1: whole serum, Serum-2: ultrafiltrated through a membrane with cut-off at molecular weight (MW) 100,000. Serum-3: through a membrane with cut-off at MW 20,000, and Serum-4: through a membrane with cut-off at MW 5,000. The infusion edema model was utilized for infusion of autologous serum into the brain. The brain tissue was histologically evaluated. The level of glutamate, total protein, and albumin was also measured in the sera used for infusion. The following results were obtained: 1) CA1 neurons were more vulnerable to all infused sera than striatal neurons, 2) there was a strong cellular response in the striatal site of infusion, but only a minimal in the CA1-sector, 3) the severity of damage of CA1 correlated with the glutamate concentrations of the infused sera, 4) further, there was a relationship between the degree of striatal neuronal loss and the amount of protein and albumin present in the infusate. It is, therefore, concluded that the neurotoxic properties of vasogenic edema fluid are also affected by specific features of the brain region of its extravasation. In addition, the pathological mechanisms associated with irreversible damage of neurons might be different in the CA1-sector and the striatum.

[Highly effective preoperative intraarterial infusion chemotherapy with CDDP for progressive uterine corpus cancer with a Sister Mary Joseph's nodule].

A 57-year-old female patient complained of atypical genital bleeding and a noxious emanation from her navel. A histological examination of the uterine body and the navel area confirmed a diagnosis of adenocarcinoma. We diagnosed it as IVb stage of uterine corpus cancer with a Sister Mary Joseph's nodule. We selectively administered intraarterial injection chemotherapy (Cisplatin 120 mg, Pirarubicin 40 mg) in the uterus and navel area (three times, once every three weeks) prior to surgery. The isolated uterus showed that the cancerous tissue had been eradicated, and we judged the cancer to be grade 3 following histopathological effective grading standards. The metastasis exhibited extreme shrinkage, but affirmed changes in the tumor quality. Currently, the patient is receiving maintenance therapy of 600 mg of Hysron H, and 600 mg of UFT. There are no indications of recurrence, and the patient is progressing well.

[A case of sacral neuroblastoma in an adult successfully treated with combination chemotherapy].

A 35-year-old woman was admitted with complaints of severe posterior femoral pain and was diagnosed as having sacral neuroblastoma by tumor open biopsy. After admission, combination chemotherapy consisting of CDDP, etoposide, CPA, and THP was started intra-arterially and intravenously. After 2 courses of chemotherapy, her symptoms markedly improved and the tumor size was reduced. Now, after completion of 16 courses of chemotherapy, she is in a state of partial remission. Hereafter, we intend to reconsider the treatment strategy including surgical therapy.

Relationship between age and nephrotoxicity following single low-dose cisplatin (CDDP) injection in rats.

We studied nephrotoxicity following a single injection of cisplatin (CDDP) at low dose (1 mg/kg) in two groups of rats aged 52 weeks (adult, A group) and 9 weeks (young, Y group). Renal platinum (Pt) was detectable in both groups 3 h after the CDDP injection, and, from 6 h to 3 d after injection, its level in the A group was higher than that in the Y group. Compared with the levels in age-matched normal rats (non-treated rats examined at time zero), the plasma urea nitrogen and creatinine levels in the A group were significantly increased, beginning 3 d after CDDP injection, while those in the Y group showed little change for 10 d after injection. Beginning 3 d after CDDP injection, the level of renal metallothionein in the Y group increased, while that in the A group decreased remarkably. The renal tissue levels of the heavy metals Zn, Cu, Mn showed similar patterns. There were no significant changes in the renal lipid peroxide (LPO) level in either the A and Y group at any time measured after CDDP injection compared with the value in the respective age-matched untreated group. Morphological evaluation demonstrated degeneration of the proximal tubules in the A group 3 d after CDDP injection. These results suggested that the renal disorders observed following CDDP injection in the A group were caused by mechanisms other than LPO such as decreased tissue metabolic function associated with aging.

Renal changes of streptozotocin-induced diabetic rats fed a low-zinc diet.

The changes in kidneys of streptozotocin (STZ)-induced diabetic rats fed a low-zinc (LZ) diet were observed. Calcium deposits were detected in the LZ-diabetic groups from the 2nd to the 8th week. The deposits were mainly detected in the corticomedullary junction, and found in the tubular lumina and epithelial cytoplasm and interstitium. Tubular morphological changes, including luminary distension, epithelial flattening, and paleness of cytoplasm and nuclei, were observed near the calcium deposits in the LZ-diabetic group over the 2nd week. Moreover, at the 8th week, wedge-shaped vasogenic lesions were found on the surface of the renal cortex in the LZ-diabetic group. No changes were detected in the control for the LZ or in the diabetic group fed a standard (SC) diet. When STZ was administered, plasma glucose level in groups fed LZ or SC diet increased in the 1st week, and over the 2nd week, glucose level was maintained at more than 400 mg/dL. Glucose level of the LZ-diabetic group did not differ from that of the SC-diabetic group. However, urinary N-acetyl-beta-D-glucosaminidase activity of the LZ-diabetic group at the 8th week was significantly higher than that of the SC-diabetic group. These findings suggested that low-zinc diet hastens renal damages in diabetic rats.

Accumulation of platinum in the intervertebral discs and vertebrae of ovarian tumor-bearing patients treated with cisplatin.

Platinum was determined by the inductively coupled plasma mass spectrometry (ICP-MS) in the intervertebral discs and vertebrae of ovarian tumor bearing patients treated with cis-diamminedichloro-platinum (II) (cisplatin). Platinum was 0.05 ng/mL at the absolute detection limit, and platinum was undetectable in the intervertebral discs and vertebrae of human specimens without cisplatin treatments. On the other hand, platinum was detected in the intervertebral discs and vertebrae of patients administered cisplatin, and platinum concentration was at levels of 1.06-10.31 micrograms/g dry tissue in the intervertebral discs and 0.60-1.28 micrograms/g dry tissue in the vertebrae, respectively. The platinum level of intervertebral discs was 4.3-fold higher than that of the vertebrae. Thus, platinum accumulates greatly in the intervertebral discs and somewhat in the vertebrae after administering cisplatin to patients for therapy.

Nephrotoxicity induced by adenine and its analogs: relationship between structure and renal injury.

Twenty-four adenine analogs were administered to mice and the relationship between the structure of analogs and the occurrence of renal injury was examined. Plasma urea nitrogen (UN) and creatinine levels were measured 24 h after oral administration of analogs. Both levels increased in the adenine-, 8-azaadenine-, isoguanine-, or 6-dimethyl aminopurine (6-DMAP)-administered group, but did not increase in the other analog groups. From light microscopy, the damages of tubuli, mainly of proximal tubuli, were observed in the kidneys of these four groups. The common property of these compounds is the strong basicity of nitrogen which binds the 6-position of the purine ring. Furthermore, UN and creatinine increased time-dependently with intravenous administration of isoguanine. When adenine was intravenously administered, UN slightly increased at 1 h, but creatinine was unchanged. No changes were observed in the 6-DMAP- or 8-azaadenine-administered group. The basicity of nitrogen which binds to the 6-position of the purine ring is thus considered to be related to the occurrence of renal injury with oral administration, and isoguanine has high affinity with the kidney.

Nephrotoxicity induced by a single dose of adenine: effects of 4-aminopyrazolo[3,4-d]pyrimidine and allopurinol.

The effects of allopurinol and 4-aminopyrazolo[3,4-d]pyrimidine (4APP) on adenine-induced renal injury in mice were examined. Plasma urea nitrogen (UN) and creatinine levels increased after the oral administration of adenine to mice. However, plasma UN and creatinine levels decreased inversely with the dose of 4APP when a different dosage of 4APP was administered together with adenine. Yet 4APP did not have any effect on the UN or creatinine levels when 4APP was administered after adenine administration. Plasma UN and creatinine levels increased in the allopurinol-administered group as in the adenine-administered group. Moreover, from light microscopic observation by hematoxylin-eosin staining, microvacuolic changes in the proximal tubuli were detected in the mouse kidney in the adenine-administered group, and epithelial cell loss, degeneration and microvacuolic changes in the proximal tubuli were observed in the mouse kidney in the adenine-and-allopurinol-administered group. However, there were no changes in the proximal tubuli in the mouse kidney in the adenine-and-4APP-administered group. These findings suggested that 4APP inhibits the action of adenine in the mouse kidney.

Experimental quantitative evaluation of transvascular removal of unnecessary substances in brain edema fluid.

We developed a model by which the transvascular removal of unnecessary substances in brain edema fluid could be measured quantitatively and chronologically. Brain stab wounds were produced in Wistar rats by insertion of paired microdialysis probes in the unilateral caudatoputamen. Homovanillic acid (HVA) was administered by microdialysis from one probe, and the HVA clearance was measured by HPLC analysis of perfusate from the other probe. Using this model, we evaluated the site of removal and whether the removal processes were affected by anesthesia or an elevated plasma concentration of the substance. As a result, 1) Probenecid did not change HVA clearance although this inhibits HVA removal via subarachnoid vessels. Therefore, HVA removal in this model was considered mainly due to intraparenchymal transvascular efflux. 2) There was no alteration in HVA removal induced by anesthesia or intravenous HVA injection. Consequently, this efflux mechanism seems to be a rather stable protective process, and seems to play a considerable role in brain microenvironmental homeostasis.

[Studies on early stage changes of peroxide lipid in isoproterenol-induced myocardial injury].

The amount of peroxide lipid in vivo in the early stage of the experimental model of myocardial infarction in a rat induced by the administration of isoproterenol (Isp) was measured as the value of malonic dialdehyde (MDA). The model of myocardial infarction was made by giving 75 mg/kg of Isp to the rat weighing 270 +/- 10 g. After the administration of Isp, the amounts of lipid in the serum and in the myocardial tissue were measured, and a blood chemistry test (glutamic oxaloacetic dehydrogenase, glutamic pyruvic transaminase, lactate dehydrogenase, free fatty acid, creatine kinase) was simultaneously carried out on the serum. The value of the amount of peroxide lipid in the serum began to elevate 3 h after the administration of Isp and reached a maximum value at 6 h. The value of the amount of peroxide lipid in the tissue began to elevate 30 min after the administration and reached a maximum at 3 h. Each blood chemistry disclosed the elevation 30 min after the administration. As mentioned above, the production of peroxide lipid in vivo on the myocardial disorder in the early stage after the administration of Isp and the biochemical changes showed a significant correlation. From these results it is suggested that the myocardial disorder induced by the administration of Isp has already developed at 30 min after the administration.

Effects of osmolality on water, electrolytes and VFAs absorption from the isolated ruminoreticulum in the cow.

Effect of the osmolality on the absorption of water, electrolyte and VFAs from the isolated ruminoreticulum under normal feeding condition were investigated in a series of the study to evaluate the rumen as a potential site of absorption in oral fluid therapy of adult cattle. Thirty of 40 l of the test solutions with varying osmotic pressure (100, 200, 300 and 500 mosmol/L, pH 6.8) were prepared using different concentrations of electrolytes and VFAs. These were infused into the isolated and emptied ruminoreticulum, and the absorption rate of water and each components were studied for 3 hrs. Marked absorption of water was observed with solutions more hypotonic than rumen fluid, the extent of which was more extensive with less osmotic pressure; the absorption rate as high as 47.6% was obtained with a solution 100 mosmol in osmotic pressure. When hypertonic solution (500 mosmol/L) was infused, however, water was transported on the contrary from the blood to the rumen. Absorption rates of electrolytes such as Na, K and Cl were increased according to the elevation of osmolalities and their concentrations in the test solutions. VFAs were also absorbed in large quantities (23.9-74.5%) in any test solutions, though the absorption rates were significantly decreased with the elevation of osmolalities. These results may indicate that the ruminal wall has a high absorptive function for water, electrolytes and VFAs when the osmolalities and the concentration of solutes in the ruminal fluid are maintained within a certain range. Furthermore, it is thought that they may work as a rational support for a possible oral fluid therapy even in adult cows.

Dynamics of ischaemic brain oedema fluid: 'dense tracer distribution around the focus', evaluated by peroxidase injection in the rat cortex.

Experimental cerebral ischaemia was produced by microembolization in rats. HRP (horseradish peroxidase) was then injected into the cerebral cortex and the extracellular fluid dynamics were observed. In the control group, there was a basic pattern in the migration of the tracer. In the ischaemic group, a 'dense tracer distribution around the focus' was found in addition to the basic pattern. This phenomenon could be observed from the second day after the ischaemic insults. The perivascular DAB reaction was marked, particularly around the focus. There was a vesicular uptake of the tracer in the vascular endothelium. This phenomenon appeared almost simultaneously with the active resolution of the oedema fluid. In addition, frequent mitotic figures were seen around the focus. It was thus possible to hypothesize that the 'dense tracer distribution around the focus' appeared at the site of active tissue reaction and had a close relationship with oedema resolution.