A model for ZnII-containing-beta-lactamase: synthesis, X-ray crystal structure of a zinc(II) complex bearing thiol group and hydrolysis of phosphate diester.

A novel N3S1 typed tripodal ligand bearing an SH group, N-(mercaptoethyl)-di(2-pyridylmethyl)amine, DPASH, was prepared and its zinc(II) complex, [ZnII(DPAS)Cl], was structurally characterized by X-ray crystallography. [ZnII(DPAS)Cl] promoted hydrolysis of sodium bis(p-nitrophenyl)hydrogenphosphate), BNP-.

Distribution of osteopontin and calprotectin as matrix protein in calcium-containing stone.

We recently reported that osteopontin (OPN) and calprotectin (CPT) are present in the matrix of urinary calcium stones, and that OPN mRNA is expressed in the renal distal tubular cells. In the present study, we examined the immunohistochemical distributions of OPN and CPT in urinary stones. The stones used in this study were passed spontaneously from the upper urinary tract. One half of each of the stones was analyzed with an infrared spectrophotometer, and were shown to be comprised of calcium oxalate, calcium phosphate, uric acid and cystine. The other half of each stone was immersed in tetrasodium ethylenediamine-tetraacetate (EDTA) solution. The half-stones were embedded in paraffin and cut into 5-microm sections. The avidin-biotin-peroxidase complex technique was employed. A monoclonal antibody to human milk-derived OPN and a monoclonal antibody to human granulocyte-derived CPT were used as primary antibodies. The immunochemical study using the OPN and CPT antibodies showed positive staining of the matrix of the urinary calcium stones. The stones showed staining in two distinct zones: a core area was stained with randomly aggregated OPN and CPT, and peripheral layers were stained in concentric circles. On the basis of our observations, it is reasonable to presume that OPN and CPT play roles as the matrix in the structure of urinary calcium stones.

[Random biopsy and recurrence risk in patients with bladder cancer].

BACKGROUND: We studied the relationship between the recurrence rate of primary superficial bladder cancer and random biopsy. METHODS: We performed transurethral resection of primary superficial bladder cancer in 144 patients, and in 55 of those cases a random biopsy was also performed. The non-recurrence rate was examined in many recurrence facters. RESULTS: The non-recurrence rate in the group with random biopsy was not significantly different from that observed in the group (100 patients) without random biopsy (chi 2 test, Generalized Wilcoxon test and multivariate analysis). Because the intravesical instillation therapy is an important inhibition factor in the recurrence rate of the primary superficial bladder cancer, we further analysed the recurrence rate in two non treated groups with or without random biopsy (13 and 35 patients respectively). No significant difference in the rate of recurrence between the two groups was observed. However, we noted recurrence of bladder cancer at biopsy positions in 8/13 patients. CONCLUSION: It appears that random biopsy may have an effect on the recurrence or the implantation of tumor cells. Further studies are needed to clarify whether random biopsy is a risk factor in the recurrence rate of primary superficial bladder cancer.

[A case of uterine prolapse with hydroureter].

We report a case of complete uterine prolapse with bilateral hydroureter. The patient was a 81-year-old woman with complete uterine prolapse suffering from dysuria and urinary incontinence. Bilateral moderate hydroureter happened to be revealed by drip infusion pyelography (DIP) but blood creatinine was normal. Hydroureter was ameliorated by vaginal hysterectomy. A brief discussion and review of the literature are given.

[Clinical evaluation of pressure-flow test to male patients with micturition disturbance].

We examined pressure-flow test results in 28 male patients with micturition disturbance in whom it is difficult to determine whether bladder outlet obstruction or impaired detrusor contractility is the cause. One reason that some patients do not improve after prostatectomy is that detrusor contractility was not estimated before preoperatively. The degree of infravesical obstruction and of detrusor function were assessed by the method of Griffiths' diagram (maximum flow rate versus corresponding detrusor pressure). Three patients showed infravesical obstruction, 11 equivocal obstruction, and 14 no obstruction. No patients showed strong detrusor function, 14 normal function, and 14 weak function. Eleven patients underwent TU-procedure. Two patients with normal detrusor function and infravesical obstruction showed good postoperative improvement of uroflowmetry results, but 4 patients with weak detrusor function and no infravesical obstruction showed no improvement. Preoperative assessment of the degree of infravesical obstruction and of detrusor function by pressure-flow testing is therefore considered useful. We emphasize that preoperative evaluation of detrusor function is an important aspect of treatment of male micturition disturbance.

[Transitional cell carcinoma of the upper urinary tract following total cystectomy for bladder cancer].

From 1975 to 1990, we treated 118 patients with urinary epithelial cancer, including 100 with primary bladder cancer, 13 with primary upper urinary tract cancer, and 5 with both diseases. Thirty-five patients with primary bladder cancer underwent total cystectomy. Upper urinary tract urothelial cancer developed in 4 patients (4.0%) and was detected only after cystectomy. Three patients had multiple bladder tumors before cystectomy and recurrent tumors under long-term bladder-preserving treatment. The other patient had had cystectomy for the primary bladder lesion. Our present policy is to perform urinary cytology once a month and intravenous urography once a year in patients with bladder cancer for early detection of secondary upper urinary tract cancer.

[A clinico-pathological study of 69 cases after total cystectomy].

A clinico-pathological study was conducted on 69 patients with bladder cancer who underwent total cystectomy. The one, three and five-year actuarial survival rates for the 69 patients were 73.3%, 48.6% and 44.1%, respectively. Survival rate was not significantly associated with sex, the number of tumors or the size of tumors. The survival rate in those aged 70 years or more was slightly worse than in those who were much younger. Patients with papillary tumors had a more favorable survival rate than those with non-papillary tumors but we could find no significant difference between those with pedunculated tumors and those with sessile tumors. The actuarial 5-year survival rates by grade were 71.9% in G1 + G2, 22.6% in G3 and 32% in non-transitional cell carcinoma; the rates by stage were 86.5% in pTa + pT1, 85.7% in pT2, 20.8% in pT3a, 18.2% in pT3b and 0% in pT4. When the stage reached pT3a, the survival rate fell remarkably. The rate of INF alpha (93.8%) was significantly better than that of INF beta (28.1%) and INF gamma (15.2%). The rate of ly0 (76.2%) was also significantly better than that of ly1 (25.5%) and ly2 (18.8%). There was no significant difference in survival between v(-) (50.7%) and v(+) (25.9%). We could find no significant difference between patients who underwent pelvic lymph nodes dissection and those who did not.(ABSTRACT TRUNCATED AT 250 WORDS)

[Studies on metoclopramide (MCP) loading test during pregnancy and puerperium for the evaluation of lactating capacity].

To evaluate the correlation between prolactin (PRL) secreting capacity and puerperal lactation, 66 women who had delivered normally without any complications during pregnancy were studied. They were divided into two groups depending on the lactating state a month after the delivery: The breast feeding group (group A, 40 cases), and the supplementary feeding group (group B, 26 cases), respectively. 10 mg of metoclopramide (MCP) loading was carried out in the 36th week of gestation and in the third day of the puerperium, and serum levels of PRL (at 0, 30, 60 min. after the MCP loading) were measured by RIA. Serum levels of insulin, glucose, triglyceride, free fatty acid, cholesterol and lipoproteins were also examined. There was no significant change in the serum levels of the various, substances during pregnancy and puerperium. Also, no significant change could be seen in the MCP loading test in the 36th week of gestation. However, on the third day of puerperium, serum PRL after 30 min. of the MCP loading showed a significant increase in group B compared with group A (587.1 +/- 202.9 vs. 431.2 +/- 196.6 ng/ml, p less than 0.02, mean +/- standard deviation). These results suggested that the MCP loading test during the early stage of puerperium might be an important index to use in determining whether the mother was ready to breast feed.

[Comprehensive approach to clinical background and effect of bromocriptine administration in patients with endocrinological polycystic ovarian disease].

Present study was performed to elucidate the clinical features and the treatment of so-called endocrinological polycystic ovarian disease (PCO). 36 cases out of 189 infertile patients who had various ovulatory disturbances were subjected during past four years. They were selected by the definitions as follows; 1) serum LH levels greater than or equal to 30 mIU/ml and serum FSH levels less than or equal to 15 mIU/ml, 2) hyper-response of LH secretion by LH-RH (100 micrograms) loading test; maximum values greater than or equal to 250 mIU/ml. Serum androstenedione (ASD), dehydroepiandrosterone-sulfate (DHA-S), estrone (E1), estradiol (E2), progesterone (P, in the mid-luteal phase) and testosterone (T) levels were examined by RIA method for the purpose of the evaluation of the endocrinological background of the PCO in Japanese women. The same examinations were also done in 8 volunteer women who had normal ovulatory menstrual cycles for the control study. In 6 cases of the PCO patients who showed biphasic BBT charts, the endometrial biopsy was done in the mid-luteal phase at the same time of the blood sampling. And the correlation between various serum hormone levels and the endometrial morphology in the PCO patients was also discussed. Then bromocriptine (5 mg/day) was administered in these patients for more than 30 days and the effect of the treatment was investigated. Serum ASD levels in the PCO patients were significantly higher than those in the control (2.52 +/- 1.30 vs. 1.43 +/- 1.21 ng/ml, M +/- S.D., p less than 0.05), while serum E2 and P levels in the patients were significantly lower than those in the control (E2: 118.6 +/- 39.5 vs. 192.5 +/- 53.9 pg/ml, p less than 0.005. P: 7.26 +/- 5.08 vs. 124.4 +/- 4.6 ng/ml, p less than 0.005, respectively). There were no significant differences in serum levels of the other hormones. By the administration of bromocriptine, serum ASD levels decreased (1.62 +/- 1.34 ng/ml, p less than 0.05), and E2 (177.9 +/- 48.6 pg/ml, p less than 0.025) and P (11.8 +/- 4.3 ng/ml, p less than 0.005) levels increased significantly into the levels of control. Serum LH levels of the patients were also suppressed by the treatment of bromocriptine (42.5 +/- 13.7 vs. 27.4 +/- 12.0 mIU/ml, p less than 0.005), however there was no change in serum FSH levels.(ABSTRACT TRUNCATED AT 400 WORDS)

[Intended super-ovulation with HMG in the treatment of so-called functional sterility].

To investigate a new treatment for so-called functional sterility, 17 out of 456 infertile patients were treated in the past three years (age: 32.7 +/- 3.6 years old, infertile period: 5.9 +/- 1.4 years, M +/- S.D.). These patients were treated with HMG-HCG. The injection of HMG (75-450iu) was started on the 5th day of the menstrual cycle, and the follicular development was observed by ultrasonography. Then, HCG (10,000-20,000iu) was injected when 3 or 4 matured follicles (with a diameter greater than 20mm) were observed. In some cases, the endometrial biopsy was done in the mid-luteal phase and the effect of the treatment was examined morphologically. Pregnancies were observed in 6 cases (35.3%) and all of them were singletons. One of them spontaneously aborted at 9 weeks of gestation, but remaining five cases delivered at full term without any complications. The total dose of HMG was 2,116.8 +/- 843.3iu, and OHSS was observed in 5 cycles out of 31 cycles (16.1%). There was no other severe complication during the treatment. The growth of the endometrium thickened in the treated cases, but no time lag in endometrial dating could be found in the morphological study. These results indicated that the intended superovulation method for the treatment of so-called functional sterility was effective through the mechanism of the improvement of the implantation ratio.

[Effects of obesity and weight gain during pregnancy on obstetrical factors].

The relationship between the body weight or weight gain during pregnancy and various obstetrical factors was investigated in 731 patients who delivered in San-ikukai Hospital for in the year 1986. The patients were classified into three groups according to their body weight in non-pregnant states: slender (n = 214), ordinary (n = 379) and obese (n = 138), according to the standard for "The decision diagram for the estimation of obesity and emaciation in Japanese" published in 1986 by the Ministry of Health and Welfare, Japan. Each of these groups was further divided into two groups according to the degree of weight gain during pregnancy (more or less than 15kg). Then, the duration of labor, the blood loss during delivery, the birth weight, the placental weight and the obstetrical abnormalities (prolonged labor, arrested labor, blood loss of more than 500 ml, fetal distress and toxemia of pregnancy) were investigated in these groups. The blood loss, the neonatal birth weight and the placental weight in the obese groups were much greater than those in the ordinary or the slender group (p less than 0.005). However, there was no significant difference in the duration of the labor among these groups. The incidence of obstetrical abnormalities in the obese group was significantly higher than in the ordinary or the slender group (chi 2 = 4.37, p less than 0.05, chi 2 = 5.27, p less than 0.025).(ABSTRACT TRUNCATED AT 250 WORDS)

[Role of TRH or metoclopramide loading test in the diagnosis of so-called occult or latent hyperprolactinemia].

Present study was performed to investigate whether TRH or metoclopramide (MCP) loading test was useful for the diagnosis of so-called occulted or latent hyperprolactinemia (transient increase of serum prolactin levels more than 30 ng/ml during night; OHP). The circadian profiles of serum prolactin levels were examined in 31 women (age: 23-32 years old) whose BBT charts showed biphasic patterns. Blood samplings had been done every two hours through an intravenous indwelling catheter without any disturbances. And seven cases of the OHP were selected. Five cases of the control were also selected at random. Then, LH-RH (100 micrograms) and TRH (500 micrograms) loading test and LH-RH and MCP (10 mg) loading test were performed to these cases in the mid-luteal phase of the same menstrual cycle at interval of two or three days, and serum FSH, LH and prolactin levels (at 0, 30, 60, 90, 120 min. after the loading test) were determined by radioimmunoassay. Serum prolactin levels in the OHP group showed significant higher levels than those of the control from 22 to 6 o'clock (p less than 0.05-0.005). By the administration of 500 micrograms of TRH, serum prolactin levels of the OHP group increased significantly compared to those of the control at all sampling points (p less than 0.05-0.005), and also by the administration of 10 mg of MCP, the same result was obtained (p less than 0.05-0.02). The maximum peak of serum prolactin levels appeared at 30 min. after TRH or MCP loading.(ABSTRACT TRUNCATED AT 250 WORDS)

The role of transferrin and ferritin in the fetal-maternal-placental unit.

The rapidly growing human fetus requires a large supply of iron, which is obtained from the iron stores of the mother. Iron is transported from mother to fetus against a concentration gradient. The placenta plays a key role in regulating the supply of iron in the fetus. In both anemic and nonanemic patients serum ferritin levels decreased and total iron-binding capacity increased as gestation progressed. The total iron-binding capacity is higher in maternal than in umbilical cord blood at delivery; this suggests that little or no ferritin or transferrin is transferred from mother to fetus. Mother and fetus appear to have independent systems controlling iron metabolism. Transferrin was localized on the site facing the intervillous space, on the surface of the microvilli of the syncytiotrophoblasts. The removal of transferrin from the surface of the trophoblast by thiocyanate and its rebinding were demonstrated. Ferritin was shown to be present in all layers of the trophoblast and especially in the syncytiotrophoblast.

Studies on the placental transport mechanism for iron.

Transferrin (Tf) and ferritin (Fr), the proteins which participate in iron transport, were examined to determine their fluctuation during pregnancy and their localization in human placental tissue, and the iron transport mechanism between mother and fetus was discussed. The main results are as follows: Maternal serum ferritin (SFr) decreased remarkably and the maternal total iron binding capacity (TIBC) increased gradually as pregnancy progressed. Maternal serum iron (SI), SFr and TIBC at delivery were 59.8 +/- 26.2 micrograms/dl, 9.6 +/- 7.2 ng/ml and 495.2 +/- 100.3 micrograms/dl, and cord blood SI, SFr and TIBC were 161.5 +/- 42.1 micrograms/dl, 160.5 +/- 67.2 ng/ml and 177.7 +/- 33.9 micrograms/dl, respectively. Peroxidase conjugating antibody method revealed the localization of Tf on the microvillous surface of syncytiotrophoblasts, and the localization of Fr in all layers of trophoblasts, especially in the neighborhood of the surface. Ferritin content of the placenta in the 1st, 2nd and 3rd trimesters was 0.07 +/- 0.03, 0.18 +/- 0.03 and 0.25 +/- 0.09 (micrograms/mg protein), respectively. These results indicate that iron is transferred from the mother to the fetus by the placental active function, and Tf receptor and Fr on trophoblasts participate in the adequate placental iron transport.