Retinoic acid-induced monocytic differentiation of HL60/MRI, a cell line derived from a transplantable HL60 tumor.

The human promyelocytic leukemia cell line HL60 differentiates to either granulocytes or monocytes/macrophages when induced with various chemicals and lymphokines. Retinoic acid (RA) induces HL60 to differentiate to granulocyte-like cells. However, HL60/MRI cells, derived from a transplantable HL60 tumor established in athymic nude mice, differentiate to monocytoid cells when cultured with RA in vitro. HL60/MRI induced with RA are monocytes based on morphology and the expression of markers and functions specific for monocytes such as: the OKM5 monocyte-specific antigen, nonspecific esterase activity, and adhesiveness. HL60/MRI is much more sensitive to RA than is HL60. Thus, the RA concentrations that induce 50% differentiation are 0.41 nM for HL60/MRI and 37 nM for HL60, and maximum differentiation occurs at 2 days for HL60/MRI and at 4 days for HL60. While RA induces HL60/MRI to monocytoid cells, other inducers of granulocytic differentiation of HL60, such as dimethyl sulfoxide and hexamethylene bisacetamide, induce HL60/MRI to granulocytes. Furthermore, 12-O-tetradecanoylphorbol-13-acetate induces both HL60 and HL60/MRI to macrophage-like cells. The isozyme phenotypes of HL60/MRI and HL60 are identical. Cytogenetic analysis of HL60/MRI indicates that many of its normal chromosomes are triploid and that it has five abnormal chromosome markers, M1-M5, three of which, M1-M3, are seen also in HL60. This unique cell line, HL60/MRI, may be useful for studying the event(s) triggering differentiation of myelomonocytic cells and the mechanism of action of RA.

The effect of sodium thiosulfate on subcellular localization of platinum in rat kidney after treatment with cisplatin.

Female Sprague-Dawley rats were given 10 mg/kg of i.v. cis-diammine-dichloroplatinum(II) (cisplatin) simultaneously with i.v. sodium thiosulfate (STS) at a 200-fold molar excess to cisplatin, then subcellular (nuclei, mitochondria, microsomes, cytosol) distribution of platinum within rat kidney cells was determined 15 min, 1 h, 8 h and 24 h after cisplatin injection. Blood urea nitrogen levels were measured in rats treated in the same manner described above. STS was found to block cisplatin-induced nephrotoxicity. However, differences in platinum concentrations in total homogenate or each subcellular fraction between STS-treated rats and controls were not significant enough to fully account for the drastic protective effect of STS against cisplatin nephrotoxicity.

The effect of cisplatin on renal ATPase activity in vivo and in vitro.

The effect of cisplatin on ATPase activity was determined in vitro and in vivo to investigate the correlation between nephrotoxicity and the inhibition of ATPase activity by cisplatin. Purified Na,K-ATPase was preincubated for 0-240 min with cisplatin at concentrations of 50-800 microM in vitro before the determination of enzyme activity. Although ATPase activity was reduced by cisplatin, either a high concentration of cisplatin (280 microM) or a long period of preincubation (160 min) with cisplatin was required to obtain 50% inhibition of ATPase activity. Similar in vitro experiments using kidney homogenate from female Sprague-Dawley rats instead of purified Na,K-ATPase were performed. Activity of Na,K-ATPase in rat kidney homogenate was inhibited by 50% after 110 min preincubation with 800 microM cisplatin or 160 min preincubation with 400 microM cisplatin. Female Sprague-Dawley rats were given 5, 7 or 10 mg/kg of cisplatin IV and BUN level, ATPase activity and Pt concentration in kidney homogenate were evaluated 1 h, 6 h, 1 day, 3 days, and 5 days after cisplatin injection. In rats given 10 mg/kg cisplatin a significant increase of BUN was observed on days 1, 3, and 5. In rats treated with 5 or 7 mg/kg of cisplatin BUN was increased on days 3 and 5. Normal ATPase activity, however, was preserved until day 3 at all doses. The highest concentration of Pt observed in kidney tissue was 19.3 micrograms/g tissue. This value was insufficient to inhibit ATPase activity significantly in vitro. Thus, it seems unlikely that the inhibition of ATPase activity is the cause of nephrotoxicity, although cisplatin can affect ATPase activity.

Sodium thiosulfate inhibits cis-diamminedichloroplatinum (II) activity.

To elucidate the mechanisms of inactivation of cis-diamminedichloroplatinum(II) (DDP) toxicity by its antidote sodium thiosulfate (STS), we studied the effects of STS on plasma concentrations of platinum (Pt) in vivo, binding of Pt to serum protein in vitro, and uptake of Pt by bacterial cells (E. coli, WP 2 uvrA strain) or cultured mouse tumor cells (FM3A) in vitro. STS did not significantly affect either plasma levels of total Pt or non-protein-bound Pt in vivo, but did inhibit binding of Pt to serum protein and cellular uptake of Pt in vitro. These results suggest that when DDP is given in combination with STS in vivo, the binding to macromolecules and entry of DDP into the cells are prevented due to formation of the Pt-thiosulfate complex in the extracellular fluid.

Prophylactic effect of methylprednisolone against cisplatin-induced nephrotoxicity in rats.

Male Sprague-Dawley rats were intravenously injected with 6.5 mg/kg of cisplatin combined with a subcutaneous methylprednisolone (MP) injection to study the prophylactic effects of steroid hormone on cisplatin-induced nephrotoxicity. In Expt. 1, the rats were injected twice with either 10, 20 or 50 mg/kg of MP4 and 0 h prior to the cisplatin injection and in Expt. 2, they were given a single injection of 100 mg/kg of MP either 24, 4, 2 or 0 h prior to the cisplatin injection. In Expt. 1, elevations of the BUN and serum creatinine levels caused by the cisplatin were dose-dependently inhibited by the administration of MP. In the second experiment, BUN and serum creatinine levels in rats which were pretreated with MP 4 or 2 h prior to the cisplatin injection were significantly lower than those in rats which received cisplatin alone. Our preliminary results suggest that pretreatment with MP is an effective means of protection against cisplatin-induced nephrotoxicity.

Laparoscopic adrenalectomy: review of 14 cases and comparison with open adrenalectomy.

We reviewed 14 cases of laparoscopic adrenalectomy and compared the results with those of a recent series of 15 consecutive patients undergoing a traditional open adrenalectomy for a benign tumor. The laparoscopic adrenalectomy group included nine patients with primary aldosteronism, three with Cushing's syndrome, one with pheochromocytoma, and one with a nonfunctioning incidentaloma. In the patient with pheochromocytoma, a good operative field was safely obtained by a combination of pneumoperitoneum at less than 6 mm Hg insufflation pressure and the abdominal wall-lift method. In both groups, the tumors were removed successfully in all cases. Laparoscopic adrenalectomy, which required neither a large skin and muscle incision nor any resection of the ribs, offered a lower morbidity and earlier recovery in spite of the longer operation time. The most important complication observed in laparoscopic adrenalectomy was that of pneumothorax secondary to an injury of the diaphragm and pleura during the dissection of the left adrenal gland using electrocautery. However, the injury wound was small, and the pneumothorax was resolved by suturing the tear under laparoscopy. These results suggest that laparoscopic adrenalectomy is a minimally invasive alternative to traditional open adrenalectomy and thus has the potential soon to become a standard procedure for the treatment of benign adrenal tumors.

Ultrastructural alterations and DNA synthesis of renal cell nuclei following cisplatin or carboplatin injection in rats.

To clarify the difference in nephrotoxicity between cisplatin and carboplatin, ultrastructural alterations and DNA synthesis of renal cell nuclei were studied in Sprague-Dawley rats which had received intravenously either cisplatin or carboplatin at an equitoxic dose. Twelve hours after cisplatin injection, nucleolar segregation accompanied by aggregated nuclear heterochromatin was observed in the third segment of the proximal tubules. Seventy-two hours after cisplatin injection, nuclear damage was more widespread while regenerative cells were also observed. Nuclear damage was not observed in the carboplatin-treated rats. Nuclear DNA synthesis of renal cells was suppressed at 8, 12 and 24 h and was accelerated at 72 h after cisplatin injection. Carboplatin did not suppress nuclear DNA synthesis at any time. The results indicate that cisplatin, but not carboplatin, can affect the renal cell nuclei. Cisplatin-induced nephrotoxicity is related to its effects on renal cell nuclei.

Experimental studies on the pharmacokinetics and nephrotoxicity of carboplatin (cis-diammine-1, 1-cyclobutane dicarboxylate platinum II) in rats.

To study the nephrotoxicity of carboplatin (cis-diammine-1, 1-cyclobutane dicarboxylate platinum II, CBDCA), an analogue of cisplatin, we examined its pharmacokinetics and functional and histopathological changes of the kidney in rats that received i.v. injection of carboplatin. Platinum concentrations in the whole plasma rapidly decreased during the first 2 hours and was undetectable at 72 hours following the carboplatin administration. Approximately 90% of the platinum in the whole plasma was ultrafilterable during the first 30 minutes. The renal tissue concentrations of platinum rapidly decayed during the first 4 hours and then slowly declined up to 120 hours following the carboplatin administration. Platinum concentrations in the renal cortex showed higher levels than those in the renal medulla throughout the experimental period. BUN levels were within the normal range except on day 7. Serum creatinine levels remained stable and normal during the 7 days. Histopathological alterations of the renal tubules were not observed during the experimental period. These results suggest that carboplatin has less nephrotoxicity than cisplatin, because of its rapid excretion through glomerulus and less accumulation in the tubular cells.

Calcium blockers enhance cisplatin-induced nephrotoxicity in rats.

To examine the effects of calcium blockers on nephrotoxicity caused by cisplatin, the renal function and renal accumulation of Pt in Sprague-Dawley rats given 6.5 mg/kg i.v. cisplatin simultaneously with several doses of verapamil or nicardipine were evaluated. BUN, serum creatinine and kidney Pt concentrations in rats given more than 5.0 mg/kg of verapamil were significantly higher than those of the control animals injected with 6.5 mg/kg i.v. cisplatin alone, and the increase of each value was dependent upon the dose of verapamil. BUN, creatinine and kidney Pt in rats injected with more than 0.5 mg/kg i.p. nicardipine were also significantly higher than those of the controls. Calcium blockers enhanced the renal accumulation of Pt and the nephrotoxicity of cisplatin.

Clinical study of asymptomatic microscopic haematuria.

A clinical statistical survey was performed on 355 patients with asymptomatic microscopic haematuria. Urologic lesions were detected in 19.4% of the patients. Urologic lesions requiring surgical treatment were found in only two patients with bladder carcinoma and with renal calculus. With the exception of glomerulonephritis, the proportion of those over 40 years who had urologic lesions was higher. It is suggested that an initial evaluation based on excretory urography, cystoscopy and ultrasonography is more important for patients over 40 years.

Platinum accumulation in the kidney and liver following chemotherapy with cisplatin in humans.

Platinum (Pt) concentrations in the kidney and liver were determined in 12 autopsied cases following cisplatin treatment. The Pt level in the liver was generally higher than that in the kidney. Pt concentration in the liver showed a significant correlation with the total dose of cisplatin. However, no dose-dependent Pt accumulation was observed in the kidney. Kidney Pt levels were correlated with the doses of cisplatin injected within a 6-month period prior to the patient's death. It is suggested that the dose of cisplatin injected during a relatively short period before the patient's death affected the Pt concentration in the kidney.

Clinical significance of urinary enzymes and beta 2-microglobulin following ESWL.

To examine the renal damage caused by shock waves, urinary excretion of enzymes and beta 2-microglobulin were determined before and after ESWL. Urine samples were obtained from 35 patients with renal stone and 26 patients with ureteric stone treated with ESWL. Urinary lactate dehydrogenase (LDH) levels significantly increased on day 0, just after ESWL, in both groups. In the ureteric stone group the kidneys received less shock waves than in the renal stone group. Increased urinary lactate dehydrogenase was considered to have derived from erythrocytes in urine. Elevated urinary N-acetyl-beta-D-glucosaminidase (NAG) levels were also observed on day 0 after ESWL in both groups, due to unknown reasons. Indirect effect of ESWL through the sympathetic nervous system or humoral factors may contribute to increases in the urinary excretion of NAG. No significant increase was found in urinary gamma-glutamyl-transpeptidase (GGTP) levels for 5 days after ESWL. Urinary beta 2-microglobulin (BMG) levels increased on day 0 in the renal stone group alone. In our present study, the clinical significance of urinary enzymes and BMG was not well evaluated, because urinary excretion of these indicators following ESWL were transient and mild.

[Urinary N-acetyl-beta-D-glucosaminidase activity in acute pyelonephritis patients with spinal cord injuries].

The urinary N-acetyl-beta-D-glucosaminidase (NAG) activities were determined in acute pyelonephritis patients with spinal cord injuries. The urinary NAG activity was significantly elevated in 23 of 31 cases (74%) compared with normal controls. Out of 7 acute pyelonephritis patients without spinal cord injuries, 4 patients (57%) showed significantly elevated urinary NAG activities. The urinary NAG activities were within normal range in 20 patients with acute simple cystitis and 11 patients with chronic complicated cystitis. Out of 6 patients with urethritis, only one case (17%) showed a significantly higher level of urinary NAG activity. Significantly higher levels in urinary NAG activities were observed in 6 of 9 patients (67%) with acute prostatitis and 5 of 9 patients (56%) with acute epididymitis. In patients with spinal cord injuries, having frequent urinary tract infections and complicated pathophysiological conditions, urinary NAG is one of the helpful laboratory findings for the diagnosis of acute pyelonephritis.

[Bone mineral density (BMD) of the calcaneus bone in 72 dialysis patients measured by ultrasonic bone absorptiometry].

BACKGROUND: To determine what factors contribute to and change BMD in dialysis patients. METHODS: Bone parameters, namely, osteosono-assessment index (calculated by speed of sound and transmission index) were measured at calcaneus bone by ultrasonic bone absorptiometry with an Aloka Acoustic Osteoscreener (Model AOS-100). Seventy two patients were 62. 8 +/- 10.5 years of age (mean +/- SD) (range 36-81) and 70.8% were males; the patients had received dialytic therapy for 52.7 +/- 41.7 months (range 2-205). The effects of sex, age, height, weight, postmenoposal years, dialysis duration, various blood parameters, oral phosphorus binding agent (CaCO3 dosage) and oral vitamin D3 treatment (D3 dosage) on BMD were assessed statistically. RESULT: BMD in male patients were significantly higher than those in female patients (Mann-Whitney test, p < 0.01). BMD showed significant correlations with age (Spearman's correlation coefficient r = -0.384, p < 0.01), height (r = 0.479, p < 0.00001), postmenoposal years (r = -0.692, p < 0.05), dialysis duration (r = 0.250, p < 0.01), blood Ca level (r = 0.292, p < 0.05) and CaCO3 dosage, postmenoposal years and dialysis duration were the variables that significantly correlated with the BMD (multiple correlation coefficient = 0.646).

Early and quantitative detection of cisplatin-induced nephrotoxicity by measurement of plasma PSP (phenolsulfonphthalein) half-life in rats.

Renal function following cisplatin injection in rats was assessed by measuring the half-life of plasma PSP. Changes in BUN, creatinine, plasma PSP half-life and urinary N-acetyl-beta-D-glucosaminidase (NAG) levels were determined in rats treated with 6.5 mg/kg i.v. of cisplatin. The highest values of BUN and creatinine were both observed on day 5. Half-life of plasma PSP was significantly prolonged on days 2 through 5 and reached a peak value on day 3. A significant increase in urinary NAG levels appeared on day 2 and the peak value was demonstrated on day 3. BUN, creatinine and plasma PSP half-life were also determined on days 3 and 5 following several doses of i.v. cisplatin. A dose-related increase in plasma PSP half-life was observed on day 3, while dose-related increases in BUN and creatinine levels were found on day 5. The evaluation of cisplatin-induced nephrotoxicity by the plasma PSP test was clearly superior for early detection compared with conventional renal function tests using BUN and creatinine.

[Prophylactic effect of methylprednisolone on cisplatin-induced nephrotoxicity in rats].

Cis-diamminedichloroplatinum (II) (cisplatin) is a very effective antitumor drug, but It has an adverse effect of nephrotoxicity. There are many reports concerning protective agents against cisplatin nephrotoxicity, but clinically useful modalities are very few. In the present study, prophylactic effects of methylprednisolone (MP) on cisplatin-induced nephrotoxicity were examined. Male Sprague-Dawley rats were injected iv with 6.5 mg/kg of cisplatin combined with a subcutaneous MP given sc in various doses at various timing. Rats were killed 5 days after cisplatin injection to determine BUN and serum creatinine levels. BUN and serum creatinine levels in rats pretreated with MP 4 or 2 hr before cisplatin injection were significantly lower than those in rats received cisplatin alone. Our preliminary results suggest that pretreatment with MP is an effective means of protection against cisplatin-induced nephrotoxicity.

Genetic and epigenetic alterations on the short arm of chromosome 11 are involved in a majority of sporadic Wilms' tumours.

Wilms' tumour is one of the most common solid tumours of childhood. 11p13 (WT1 locus) and 11p15.5 (WT2 locus) are known to have genetic or epigenetic aberrations in these tumours. In Wilms' tumours, mutation of the Wilms tumour 1 (WT1) gene at the WT1 locus has been reported, and the WT2 locus, comprising the two independent imprinted domains IGF2/H19 and KIP2/LIT1, can undergo maternal deletion or alterations associated with imprinting. Although these alterations have been identified in many studies, it is still not clear how frequently combined genetic and epigenetic alterations of these loci are involved in Wilms' tumours or how these alterations occur. To answer both questions, we performed genetic and epigenetic analyses of these loci, together with an additional gene, CTNNB1, in 35 sporadic Wilms' tumours. Loss of heterozygosity of 11p15.5 and loss of imprinting of IGF2 were the most frequent genetic (29%) and epigenetic (40%) alterations in Wilms' tumours, respectively. In total, 83% of the tumours had at least one alteration at 11p15.5 and/or 11p13. One-third of the tumours had alterations at multiple loci. Our results suggest that chromosome 11p is not only genetically but also epigenetically critical for the majority of Wilms' tumours.

Reproduction of sound from old disks by the laser diffraction method.

A laser diffracton method was developed as a noncontacting nondestructive means of reproducing the sound from old disk-type records. In the method, the sound signals encoded in the sound grooves are converted to the direction of the linear structure in the diffraction patterns which are produced by the reflection of a laser beam from the sound grooves. The tracking is performed using a variation of the intensity distribution of the diffraction pattern when the sound grooves deviate from the illuminating spot. The properties of the reproduced sound are discussed in comparison with those reproduced by the laser-beam reflection method [Appl. Opt. 25, 597 (1986)] developed for wax phonograph cylinders and by traditional phonographs.

First-order intensity and phase statistics of Gaussian speckle produced in the diffraction region.

General expressions for first-order statistics of Gaussian speckle produced in the diffraction region are derived for an arbitrary profile of the illuminating beam with a plane wave front. The statistical properties of the complex amplitude, the intensity, and the phase of speckles are studied under illumination of a Gaussian beam. The joint probability density function of the speckle field characterized by an equiprobability density ellipse is investigated in some detail with an intimate relation to the correlation coefficient between the real and imaginary parts of the complex speckle amplitude. This correlation coefficient is found to affect appreciably the statistics of speckles produced especially in the near-field diffraction region when a standard deviation of the random phase variation produced by a diffuse object under illumination is relatively small and the number of independent scatterers contributing to the formation of speckles is small.