Yukimichi Kawada

Chemopreventive effects of diosmin and hesperidin on N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine‐induced urinary‐bladder carcinogenesis in male ICR mice

The chemopreventive effects of 2 flavonoids (diosmin and hesperidin) on N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (OH‐BBN)‐induced urinary‐bladder carcinogenesis were examined in male ICR mice. Animals were divided into 11 groups, and groups 1 to 7 were given OH‐BBN (500 ppm) in the drinking water for 6 weeks. Groups 2 to 4 were fed diets containing the test compounds (group 2, 1000 ppm diosmin; group 3, 1000 ppm hesperidin; group 4,900 ppm diosmin + 100 ppm hesperidin) for 8 weeks during the initiation phase, while groups 5 to 7 were fed these diets, respectively, for 24 weeks during the post‐initiation phase. Groups 8 to 11 were controls, given only the test compounds or untreated basal diets throughout the experiment (weeks 1 to 32). The incidence of bladder lesions and cell‐proliferation activity estimated by enumeration of silver‐stained nucleolar‐organizer‐region‐associated proteins (AgNORs) and by the 5‐bromodeoxyuridine (BUdR)‐labeling index was compared among the groups. Feeding of the test compounds, singly or in combination, during both phases caused a significant reduction in the frequency of bladder carcinoma and pre‐neoplasia. Dietary administration of these compounds significantly decreased the AgNOR count and the BUdR‐labeling index of various bladder lesions. These findings suggest that the flavonoids diosmin and hesperidin, individually and in combination, are effective in inhibiting chemical carcinogenesis of the bladder, and that such inhibition might be partly related to suppression of cell proliferation. Int. J. Cancer 73:719–724, 1997.

Clinical Evaluation of Serum Prostate‐Specific Antigen‐Alpha1 ‐ Antichymotrypsin Complex Values in Diagnosis of Prostate Cancer: A Cooperative Study

Background We studied the clinical significance of serum prostate‐specific antigen bound to α1‐antichymotrypsin (PSA‐ACT) values determined with a newly developed enzyme immunoassay.

Nucleolar organizer regions in rat urinary bladder tumors induced by N-butyl-N-(4-hydroxybutyl)nitrosamine

SummaryThe number of nucleolar organizer regions (NORs) stained by the one-step silver colloid method was measured in preneoplastic and neoplastic bladder lesions induced by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in rats. Male ACI/N rats, 6 weeks of age, were given 0.05% BBN in drinking water for 5, 8, 12, 18 and 30 weeks to induce preneoplastic and neoplastic transitional cell lesions. The mean numbers of silverstained NORs (AgNORs) in such lesions were as follows: untreated transitional epithelium (n = 6), 1.26 ±0.09; transitional cell epithelium outside focal lesions (n= 10), 1.75 ±0.10; simple hyperplasia (n= 10), 2.01 ±0.15; papillary or nodular (PN) hyperplasia (n= 10), 2.15±0.19; transitional cell papilloma (n= 5), 2.37 ±0.12; transitional cell carcinoma (n= 5), 3.52 ±0.23. Thus, the mean number of AgNORs showed a step-wise increase from untreated and treated, histologically normal transitional epithelium through simple hyperplasia and PN hyperplasia to transitional cell papilloma and carcinoma. These results suggest that the mean number of AgNORs may reflect the proliferative nature of bladder lesions induced by BBN, as reported in preneoplastic and neoplastic lesions in other organs. PN hyperplasias were classified into two types based upon the mean number of AgNORs, indicating that they include reversible and irreversible changes in contrast with simple hyperplasia which is reversible change.

Studies on the Phagocytic function of Urinary Leukocytes

Urine specimens from patients with urinary tract infection (UTI) were examined to determine the rate of phagocytosis and viability of urinary leukocytes. The phagocytic function of urinary leukocytes was also studied in vitro. The mean rate of viable urinary leukocytes was 83 per cent and the phagocytic potency was confirmed by light and electron microscopic studies. In 99 per cent of 113 patients with UTI, urinary leukocytes were shown to have phagocytized bacteria. The rate of phagocytosis in chronic UTI was higher than that in acute UTI. Urinary osmotic pressure and the positive or negative of antibody coated bacteria were supposed to be factors influencing phagocytic potency of urinary leukocytes.

Clear Cell Melanoma of the Renal Pelvis Presenting as a Primary Tumor

TO date only 3 cases of malignant melanoma presenting as a renal pelvic mass have been reported in adulthood.1.2 To our knowledge we report the first primary malignant melanoma composed of clear cells in the renal pelvis of a child. CASE REPORT A 3-year-old girl was found to have microscopic hematuria during a routine medical examination. Excretory urography revealed a filling defect in the left renal pelvis. Abdominal sonography showed a 2 cm. isoechoic tumor with normal renal parenchyma within the lesion. Abdominal computerized tomography confirmed the lesion in the renal pelvis and demonstrated no evidence of visceral metastasis (fig. 1). Left radical nephrectomy was performed for the presumed diagnosis of Wilms tumor. Histological evaluation of the surgical specimen revealed a solid tumor composed of clear cells with focal pigmentation in the renal pelvis. The transitional cell epithelium was intact (fig. 2). Fontana-Masson staining was positive for melanin and tumor cells were immunoreactive for HMB-45. The patient had no history of malignant melanoma and postoperative clinical and radiological evaluation failed to reveal any other focus of melanoma. Microscopic hematuria resolved postoperatively. The patient remained asymptomatic and without evidence of residual disease 13 months after surgery. DISCUSSION

Significance of AgNOR counts for distinguishing carcinoma from adenoma and hyperplasia in parathyroid gland

Nucleolar organizer region proteins, which can be stained and visualized by an argyrophil technique (AgNORs), are markers of cell activities, such as DNA transcription and proliferation, and they are useful for differential diagnosis between benign and malignant tumors. We counted both AgNOR numbers in 25 parathyroid lesions (three carcinomas, 11 adenomas, 10 hyperplasias, and one hyperplasia with carcinoma) to determine if the AgNOR number could be useful as a diagnostic aid in parathyroid neoplasms and hyperplasias, because it is often difficult to histopathologically distinguish among these lesions. The AgNOR numbers were significantly higher in carcinomas (3.18 +/- 0.05) than in adenomas (1.67 +/- 0.30, P < .001) or hyperplasias (1.85 +/- 0.16, P < .001), but there was no significant difference between adenomas and hyperplasias. These results suggest that AgNORs may be useful as an adjunct to discriminating carcinomas from adenomas or hyperplasias in the parathyroid gland.

IN VITRO SELECTION OF FLUOROQUINOLONE-RESISTANT NEISSERIA GONORRHOEAE HARBORING ALTERATIONS IN DNA GYRASE AND TOPOISOMERASE IV

PURPOSEnWe attempted to select increasingly fluoroquinolone-resistant strains of Neisseria gonorrhoeae in vitro and to assess whether selected mutants harbored alterations in the GyrA subunit of DNA gyrase and the ParC subunit of DNA topoisomerase IV, which were analogous to those in fluoroquinolone-resistant clinical isolates.nnnMATERIALS AND METHODSnA fluoroquinolone-susceptible strain was exposed to norfloxacin in vitro. Selected mutants were sequentially exposed to norfloxacin, and this procedure was repeated. For 11 mutants, minimum inhibitory concentrations (MICs) of antimicrobial agents were determined, and mutations in the region corresponding to the quinolone resistance-determining region (QRDR) of the Escherichia coli gyrA gene and the analogous region of the parC gene were analyzed.nnnRESULTSnMutants obtained in one step exhibited significantly increased MICs of norfloxacin, ofloxacin and ciprofloxacin and had a single amino acid change in GyrA. Two-step mutants exhibited significantly higher norfloxacin MICs. Three of four two-step selected strains had single amino acid changes in both GyrA and ParC. Three-step mutants exhibited further increases in fluoroquinolone MICs and were assigned to the ciprofloxacin-resistant category. Two had a double amino acid change in GyrA, and one had a double GyrA change and a single amino acid change in ParC.nnnCONCLUSIONnWe selected fluoroquinolone-resistant strains that carried GyrA and ParC alterations analogous to those in clinical isolates. The serial accumulation of changes in the QRDR of GyrA and the analogous region of ParC was associated with a stepwise increase in fluoroquinolone resistance, although the development of additional alterations in other regions of GyrA and ParC or other mechanisms of fluoroquinolone resistance also might contribute to the enhancement in fluoroquinolone resistance. The clinical emergence of fluoroquinolone-resistant strains may be due to in-vivo stepwise selection of strains with genetic alterations in GyrA and ParC, as observed here in the in-vitro selection of fluoroquinolone-resistant mutants.

HLA-A and B antigens in patients with Peyronie disease

Using microdroplet lymphocyte cytotoxicity test, as a method of HLA typing, 9 patients with Peyronie disease were studied from a viewpoint of relationship between this disease and HLA-A and B antigens. The frequencies of each antigen in the patients were compared with those of healthy controls. No significant difference of distribution in HLA antigens was observed between the patients and the control group.

Comparison of in vitro antimicrobial activity of AM-1155 with those of tosufloxacin and fleroxacin against clinical isolates of Neisseria gonorrhoeae harboring quinolone resistance alterations in GyrA and ParC.

The in vitro antimicrobial activities of AM-1155, a new fluoroquinolone, tosufloxacin and fleroxacin were tested against 55 clinical isolates of Neisseria gonorrhoeae using the agar dilution method. In our previous study, all the strains had been examined for mutations in the region corresponding to the quinolone-resistance determining region of the Escherichia coli gyrA gene and the analogous region of the parC gene, and tested for susceptibility to ciprofloxacin. In this study, the 55 isolates of N. gonorrhoeae were assigned to one of three categories based on the presence or absence of alterations in GyrA and ParC. In each category, the antimicrobial activity of AM-1155 against the isolates was compared with those of tosufloxacin and fleroxacin. The MICs of AM-1155 for 11 highly fluoroquinolone-resistant isolates with alterations in both GyrA and ParC ranged from 0.06 to 1.0 microgram/ml. The MICs inhibiting 50% (MIC50) and 90% (MIC90) of these isolates were 0.125 and 1.0 microgram/ml, respectively. The MICs of AM-1155 for 20 moderately fluoroquinolone-resistant isolates with alterations only in GyrA ranged from 0.03 to 0.25 microgram/ml (MIC50, 0.06 microgram/ml; MIC90m, 0.125 microgram/ml). The MICs of AM-1155 for 24 of the quinolone-susceptible isolates without alterations in either GyrA or ParC ranged from 0.004 to 0.03 microgram/ml (MIC50, 0.008 microgram/ml. MIC90, 0.015 microgram/ml). There were significant differences between the MIC distribution of AM-1155 and each corresponding MIC distribution of tosufloxacin and fleroxacin in these three categories to which the 55 isolates were assigned (p < 0.05). Based on the MIC90S of the tested fluoroquinolones, AM-1155 was two- and eightfold more active against the highly fluoroquinolone-resistant isolates than tosufloxacin and fleroxacin, respectively. Against the moderately fluoroquinolone-resistant isolates, AM-1155 was four- and sixteenfold more active than tosufloxacin and fleroxacin, respectively. Against the quinolone-susceptible strains, AM-1155 was also two- to fourfold more active than the other fluoroquinolones. Overall, AM-1155 exhibited more potent in vitro activity against both quinolone-resistant and quinolone-susceptible isolates of N. gonorrhoeae than tosufloxacin and fleroxacin. In ciprofloxacin treatment failures of gonorrhea at single doses of 500 mg. MICs for the causative organisms have ranged from 1.0 to 16.0 micrograms/ml. The MICs of AM-1155 for the isolates harboring quinolone resistance-associated genetic alterations, including strains exhibiting ciprofloxacin MICs of 2.0 and 8.0 micrograms/ml, still ranged from 0.03 to 1.0 microgram/mL A single-dose study in humans has demonstrated higher peak serum concentrations and longer half-lives of AM-1155, resulting in the AUC0-00 values of AM-1155, which are threefold greater than those of ciprofloxacin at the single doses of 400 and 600 mg. Because of its potent in vitro antimicrobial activity and advantageous pharmacokinetic behavior, AM-1155 may be a clinically useful agent for treating gonorrhea including that caused by quinolone-resistant strains.

Fluoroquinolone Associated Bladder Stone

Fluoroquinolones have a broad spectrum of antimicrobial activity and they have been shown to be highly effective in curing urinary tract infection. Tosufloxacin is a fluoroquinolone developed in Japan. We report on a patient in whom tosufloxacin was associated with the formation of bladder stones. CASE REPORT