Properties of glucocorticoid receptors in Epstein-Barr virus-transformed lymphocytes from patients with familial cortisol resistance.

In a previous report of two patients with familial glucocorticoid resistance due to reduced numbers of glucocorticoid receptors (GR), we have shown decreased numbers of GR in peripheral mononuclear cells and cultured fibroblasts but normal affinity of GR in both patients. In this study, peripheral lymphocytes from these patients, one patient's son and daughter, and normal subjects were transformed with Epstein-Barr virus. Reduced numbers and normal affinity of GR were found in the Epstein-Barr virus-transformed lymphocytes from both patients while the son and daughter had normal numbers and affinity of GR. The thermal stability of GR and thermal activation of cytosolic receptors in both patients were found to be normal. Although the percentages of nuclear bound GR were similar in both patients and normal controls, the absolute amounts of nuclear bound GR of the patients were about one-half that of normal controls. These abnormal properties of GR (reduced numbers of GR) were preserved in the transformed cells from the patients.

Inhibitory effect of somatomedin C/insulin-like growth factor I on adrenocorticotropin- or forskolin-induced steroidogenesis in isolated rat adrenocortical cells.

The effects of recombinant human somatomedin C/insulin-like growth factor (IGF-I) on the steroidogenic response of isolated rat adrenocortical cells to ACTH, forskolin, and (Bu)2cAMP were examined during short term incubations. The effect of IGF-I on cAMP production by cells stimulated with ACTH or forskolin was also examined. IGF-I inhibited ACTH-, forskolin-, and (Bu)2cAMP-induced corticosterone production in a concentration-dependent manner. IGF-I (30 ng/ml) also significantly inhibited ACTH-induced cAMP production. However, the peptide had no significant effect on forskolin-induced cAMP production. IGF-I suppressed ACTH-induced cAMP production both in the presence and absence of 3-isobutyl-1-methylxanthine, suggesting that IGF-I acts to inhibit the formation of cAMP rather than the stimulation of cAMP degradation. The observation that IGF-I inhibited steroidogenesis induced by (Bu)2cAMP strongly suggests that one site of inhibition is at some step(s) distal to cAMP formation. However, the inhibition of cAMP production after stimulation with ACTH also suggests a plasma membrane site of action for IGF-I in adrenocortical cells.

Augmented induction by dexamethasone of metallothionein IIa messenger ribonucleic acid in fibroblasts from a patient with cortisol hyperreactive syndrome.

Northern blot analysis was used to investigate the effect of dexamethasone (Dex) or zinc on messenger RNA (mRNA) levels of metallothionein-IIa (MT-IIa) in fibroblasts from a patient with cortisol hyperreactive syndrome and from three normal subjects. Dex was seen to increase MT-IIa mRNA levels and brought them to a maximum after 12 h. Zinc also increased the levels of MT-IIa mRNA and brought them to a maximum at 8 h after the addition. Dex as well as zinc caused a dose-related increase in MT-IIa mRNA levels. Dex had a maximal inductive effect on MT-IIa at a concentration of 10(-6) mol/L and zinc at a concentration of 10(-4) mol/L. There was no significant difference in the levels of basal expression of the MT-IIa gene between the patient's and normal fibroblasts. But in three separate experiments induction of MT-IIa gene by Dex obtained for the patient's fibroblasts was almost twice as much as that for normal fibroblasts. On the other hand, there were no significant difference in induction by zinc between the patient's and normal fibroblasts. These data indicated that the patient's cells were hyperreactive to glucocorticoids as seen from the effect of Dex on the MT-IIa mRNA levels.

Quantitative comparison of aromatase induction by dexamethasone in fibroblasts from a patient with familial cortisol resistance and a patient with cortisol hyperreactive syndrome.

The ability of dexamethasone to induce aromatase activity was tested in fibroblasts from a patient with familial cortisol resistance, a patient with cortisol hyperreactive syndrome, and five normal subjects. Dexamethasone increased enzyme activity in all cases in a concentration-dependent manner (over a range of 1-1000 nmol/L). In fibroblasts from a patient with familial cortisol resistance, the response curve of dexamethasone-induced aromatase activity was shifted to the right compared to that of normal cells. However, the maximal induction of the enzyme at 1 mumol/L dexamethasone was unchanged in cortisol-resistant fibroblasts. On the other hand, in fibroblasts from the patient with the cortisol hyperreactive syndrome, the half-maximal effect of dexamethasone was similar to that in normal cells, but maximum induction of aromatase activity was 2 times greater than that in normal cells. The glucocorticoid antagonist RU 486 inhibited dexamethasone-induced aromatase activity in these patients' cells and in normal cells in a concentration-dependent manner, indicating that the altered effects of dexamethasone on aromatase induction observed in each cell type were mediated through glucocorticoid receptors.

A patient with hypocortisolism and Cushing's syndrome-like manifestations: cortisol hyperreactive syndrome.

One patient is reported who has the manifestations of Cushing's syndrome in spite of persistent hypocortisolemia. His serum levels of cortisol and free cortisol were below normal, and 24-h urinary excretion of 17-hydroxycorticosteroids and cortisol were decreased. There was a rapid and substantial increase in serum cortisol in response to synthetic ACTH-(1-24). Plasma levels of ACTH were marginally increased by successive administration of CRH and vasopressin, which were followed by substantial increases in serum cortisol. Glucocorticoid activity of the patient's serum, as measured by a RRA was low. There were no responses of urinary 17-hydroxycorticosteroids after metyrapone treatment. These laboratory examinations ruled out any known clinical conditions resulting in hypocortisolemia. The clinical condition could also be explained by cortisol hyperreactivity of the patient's cells. In vitro hyperreactivity to glucocorticoids was demonstrated in cultured skin fibroblasts whose aromatase activity was increased 1.5- to 1.8-fold above that of normal cells, and [3H]thymidine incorporation was inhibited more effectively by the addition of cortisol or dexamethasone. The mechanism by which the patient is hyperreactive to glucocorticoids remains unexplained.

Analysis of retrograde ejaculation using color Doppler ultrasonography before and after transurethral collagen injection.

Transurethral bladder neck collagen injection therapy was performed in a patient with retrograde ejaculation. The phenomenon of retrograde ejaculation and its correction after the therapy were clearly demonstrated by color Doppler ultrasonography. To our knowledge this is the first report showing successful observation of retrograde ejaculation using color Doppler ultrasonography.

Steroidogenic effect of ent-kaur-16-en-15 beta-ol (kaurenol) on isolated rat adrenal cells.

In an in vitro bioassay system for adrenocorticotropic hormone using isolated rat adrenal cells, kaurenol, a diterpene alcohol, stimulated corticosterone production and augmented the steroidogenic effect of adrenocorticotropin or forskolin, dose-dependently. Kaurenol had no effect on cyclic AMP production by the cells. The diterpene also had no stimulatory effect on the adrenal adenylate cyclase activity in a cell free system. The results suggest that this particular diterpene exerts a steroidogenic effect through a mechanism independent of cyclic AMP generation.

Induction of differentiation of human phaeochromocytoma cells in culture by epidermal growth factor and insulin.

A primary culture of malignant phaeochromocytoma cells obtained from a left adrenal tumor of a 56 year old woman complaining of occasional palpitation and headache was established. The addition of epidermal growth factor (EGF) and insulin to the culture medium induced a profound development of network formation of axon-like processes. At the same time, the secretion of catecholamines from cultured cells was also increased. EGF and insulin were demonstrated to induce the differentiation of malignant phaeochromocytoma cells in primary culture.

Rounding of cultured human carcinoid tumor cells by forskolin.

A primary culture of carcinoid cells obtained from a metastasized brain tumor of a 30 year old man was established. Rounding of carcinoid cells was induced by the addition of 20 microM forskolin or 1 mM dibutyryl cyclic AMP to the culture medium. The cyclic AMP content in carcinoid cells was increased thirtyfold by the addition of 20 microM forskolin. The results provided evidence that cyclic AMP might also be involved in rounding of human malignant carcinoid cells.

1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging ability of sake during storage.

A quantitative method of analyzing the ability of sake to scavenge 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) was established and applied to sake during storage. The DPPH-scavenging ability of sake was found to decrease non-enzymatically dependent on the length and/or temperature of storage, but the decrease gradually leveled off. From the rate constant of the initial decrease in DPPH-scavenging ability, the Arrhenius activation energy was calculated to be 15.8 kcal/mol. The decrease in the DPPH-scavenging ability of sake is considered to be attributable to the generation of free radicals. These findings suggest that DPPH-scavenging ability can be utilized as an index to describe the sensory maturation of sake that will complement other maturation indexes such as 3-deoxyglucosone and Absorbance (420 nm). The decrease in the DPPH-scavenging ability of sake correlated significantly (p < 0.01) with sensory evaluations of the maturation.

Prostatic malacoplakia: a case report with a review of 49 cases of malacoplakia of various sites in Japan.

We reported a 62-year-old man with malacoplakia of the prostate, and reviewed 49 cases of malacoplakia hitherto observed in Japan in which the lesions originated from the urogenital tract, except for one gastric case. E. Coli was emphasized as a possible causative agent for malacoplakia especially in the urogenital tract. The possible histiocytic origin of von Hansemann cells was stressed by demonstrating cytoplasmic processes and desmosomes in our prostatic case. An adjuvant use of cholinergic agents and ascorbic acid with chemotherapeutic agents was recommended for treating malacoplakia.

[Drug sensitivity and expression of beta-lactamase in enterococci isolated at Okayama University Hospital].

The following results were obtained from a study of Enterococci isolated at the Okayama University Hospital from 1990 to 1994. 1) The isolation frequency of Enterococci was about 6% for each year. 2) The percentage of Enterococcus faecalis (E. faecalis) among Enterococci decreased each year, but the percentage of Enterococcus faecium (E. faecium), and Enterococcus avium (E. avium) increased each year. 3) Results of drug sensitivity tests revealed that the incidence of ofloxacin (OFLX) resistant E. faecalis and E. avium increased each year. Also, the resistance of E. faecalis to ampicillin (ABPC), the primary medication choice for treating E. faecalis, increased each year. 4) A few vancomycin (VCM) non-sensitive Enterococci were isolated. These bacteria showed low-sensitivity toward ABPC, imipenem (IPM), gentamicin (GM), and OFLX, and the extent of multiple drug resistance increased each year. 5) One hundred strains of Enterococci were selected to examine the expression of beta-lactamase using the broth method and the iodine-starch method. However, beta-lactamase producing strains were not detected.

[Pharmacokinetics of arbekacin in healthy volunteers and patients with renal insufficiency].

Pharmacokinetics of arbekacin (HBK), a new aminoglycoside, was studied. Serum concentrations and urinary excretion were determined after single intravenous drip infusion of 100 mg HBK for 1 hour to healthy volunteers and patients with renal insufficiency of various kinds. The drug concentration was determined with bioassay, fluorescence polarization immunoassay (FPIA) and high-performance liquid chromatography (HPLC). Pharmacokinetic analysis was made in accordance with the two-compartment open model, and 24-hour endogenous creatinine clearance (Ccr) was used as the renal function index. In all cases peak serum levels were detected 1 hour after administration, and similar values were noted regardless of subjects' proficiencies of renal function. However, the serum clearance during beta-phase tended to be prolonged parallel with the degree of renal insufficiency. The excretion of HBK into urine was prolonged and cumulative recovery tended to be decreased in association with the decreased valued of Ccr.

[Clinical study of ofloxacin in complicated urinary tract infection].

Ofloxacin (OFLX) was administered at a dose of 200 mg, three times daily (600 mg) for 14 days, to 15 patients suffering from complicated urinary tract infection. According to the criteria for clinical evaluation by the UTI committee, excellent and moderate responses were seen in 54% at 5 days evaluation, and in 91% at 14 days evaluation. No serious side effects were recognized and no changes in laboratory examination data were observed. We conclude that OFLX was useful in treatment of urinary tract infection.

[Treatment of advanced bladder cancer with intra-arterial infusion of cisplatinum (CDDP) and aclacinomycin (ACR), combined with angiotensin II].

Ten patients with advanced bladder cancer were treated with intra-arterial infusion therapy. The patients consisted of nine males and one female between 55 and 82 years old (median: 70 years). In all patients, cisplatinum (CDDP) (2 mg/kg), aclacinomycin (ACR) (0.5 mg/kg) and Angiotensin II (25 mg) were infused via the internal iliac artery for a period of about 30 minutes. Seven patients also received X-ray therapy with a linac. The efficacy of this therapy was assessed by computed tomographic scanning, sonography and cystoscopy. As a result of this assessment, 2 patients were rated complete response "(CR)", 6 partial response (PR) (showing 50% or more reduction in the lesion) and 2 no change "(NC)". To compare the efficacy of this therapy for two histopathologically defined groups of patients (patients with grades 2 and 3 cancer), one patient was rated "CR", four "PR" and two "NC" in the grade 3 group (total 7 patients), while one was rated "CR" and two "PR", in the grade 2 group (total 3 patients). In effective cases, pollakiuria and miction pain disappeared shortly following intra-arterial infusion therapy. As for side effects of the therapy, mild nausea or vomiting was observed in all patients, while leukopenia was noted in one patient.

[Prostatic tissue and fluid levels of lomefloxacin (NY-198)].

The concentration of Lomefloxacin (NY-198), a new quinolone, in human prostatic tissue and fluid was examined. Lomefloxacin was concentrated in prostatic gland and the mean ratios of prostatic tissue levels to serum levels (P/S) were 1.39-1.83 during 2 to 6 hrs after administration. Concerning prostatic fluid concentrations, there were no significant differences between healthy volunteers and patients with chronic prostatitis. The mean ratios of prostatic fluid levels to serum levels (Pf/S) were 0.36-0.77 during 1 to 4 hrs after administration. The concentration of Lomefloxacin in the prostatic tissue and fluid is high enough to eradicate the majority of pathogens causing bacterial prostatitis.

[A comparative study of a thick and standard loop in transurethral resection of the prostate].

We compared the safety and efficacy of transurethral resection of the prostate (TURP) with a thick loop and with a standard loop. We compared 36 consecutive men (median age, 70 years) with symptomatic benign prostatic hyperlasia (BPH) treated by TURP with a thick loop to a cohort of 36 men (median age, 72 years) treated by TURP with a standard loop. The safety parameters of evaluation included the operative time, blood loss, chronological changes in serum sodium, and complications. The efficacy parameters of evaluation included International Prostate Symptom Score, quality of life assessment, peak urinary flow rate, and post-void residual urine volume. The operative time (median, 49.5 versus 43.5 minutes), blood loss (median, 179 versus 127 ml), and change in serum sodium (median, -4.0 versus -6.0 mEq/L) were not significantly greater in the thick loop group than in the standard loop group, respectively. There were no major complications in either group. Clinically significant improvement was observed in all efficacy parameters in both groups, with no difference between the two groups. These results suggest that TURP with a thick loop is not necessarily superior to TURP with a standard loop in terms of decreasing the blood loss and decreasing the operative time.

[Immunological and biochemical characterization of urinary carcinoembryonic antigen and its clinical significance].

The immunobiochemical properties of urinary carcinoembryonic antigen (U-CEA) purified from non-tumor-bearing patients with urinary tract infection were investigated. Moreover, immunohistochemical localization of CEA in the normal urothelium was studied and clinical significance of U-CEA was also analyzed statistically. The purified U-CEA showed a molecular weight of approximately 160,000 by the Western blotting and an antigenicity identical to the purified CEA (Calbiochem Corp.). Amino acids analysis of U-CEA revealed nearly the same values as the purified CEA or the theoretical values of CEA, except for several amino acids susceptible to hydrolysis and sugar chains. Therefore, it was assumed that the U-CEA was a new CEA-related antigen, different from nonspecific cross-reacting antigen (NCA) or even CEA itself. Immunohistochemically, CEA was found mainly at the cytoplasmic membrane in normal urothelial cells. The urine samples, collected from patients with various urological disorders, were divided into the inflammation group, showing 5 leukocytes/hpf or more in the urine, and the non-inflammation group, showing less than 5 leukocytes/hpf. The groups were subdivided and U-CEA levels were statistically studied; it was found that the U-CEA levels strongly reflected the presence of inflammation, rather than the presence of urothelial tumors. Consequently, it was considered that CEA was present in the normal urothelium and that destruction and regeneration of these cells due to tumor or inflammation might release CEA, not NCA, into the urine.

[Antimicrobial prophylaxis in transurethral resection of the prostate].

OBJECTIVE: A prospective trial was performed to propose a suitable antimicrobial prophylaxis in patients undergoing transurethral resection of the prostate (TUR-P). SUBJECTS AND METHODS: Patients who underwent TUR-P due to symptomatic prostatic hyperplasia between April 1995 and February 1996 were included. Based on the results of urinalysis obtained within preoperative 3 days, the patients were classified into Group I (less than 5 WBC/hpf and bacterial count of less than 10(4) CFU/ml in urine specimen), and Group II (5 or more WBC/hpf or bacterial count of 10(4) or more CFU/ml in urine specimen). Furthermore, each group was randomly subdivided into Group A and Group B according to the period of antimicrobial administration. As prophylactic antimicrobials, cefazolin (CEZ) was used in Group I and CEZ or cefotiam (CTM) in Group II. The antimicrobial was administered only on the day of operation in Group IA (n = 92), for 3 days in Group IB (n = 96), 2 days in Group IIA (n = 37), and 4 days in Group IIB (n = 30). On the day of operation, the antimicrobial was infused immediately before the operation. The presence or absence of pyuria, bacteriuria and febrile infection, and the period required for normalization of the urinalysis were the major points evaluated here. RESULTS: No significant differences were observed in any parameters with respect to the period of administration of antimicrobial between the groups, but in both Group I and Group II, the incidence of febrile infection was higher in the groups with shorter antimicrobial administration periods. The mean period for normalization of the urinalysis required 68.4, 68.6, 65.2 and 58.2 days in Group IA, Group IB, Group IIA and Group IIB, respectively. CONCLUSION: It is concluded that 3 or 4-day administration of first or second generation parenteral cephems is generally acceptable regimen for antimicrobial prophylaxis in patients undergoing TUR-P.

[Study on the sclerosing agent for the treatment of simple renal cysts].

BACKGROUND: The objective of this study is to evaluate the sclerosing agents for simple renal cysts. METHODS: Thirty-three renal cysts of 30 patients were treated by injection of sclerosing agents with the guidance of ultrasonography. Among them, 22 were treated with ethanol, and 11 with minocycline-hydrochloride (MINO). RESULTS: Clinical efficacy rates and volume reduction rates in each treatment group of ethanol or MINO, were 100% and 90.9%, and 91.2 +/- 13.1% (mean +/- SD) and 83.2 +/- 28.5%, respectively. Although several complications including pain and intoxication were observed in ethanol (47.4%), no complications were observed in MINO. CONCLUSION: The instillation of MINO into renal cysts is an effective, safe and simple treatment. It is thought that it can be the first choice of treatment for simple renal cysts.