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PURPOSE/OBJECTIVE(S) Stereotactic body radiotherapy (SBRT) is an emerging non-invasive treatment in the management of cardiac arrhythmias. The purpose of this research is to evaluate the MR imaging in the detection of radioablation lesion at the cavotricuspid isthmus (CTI) in mini pigs. MATERIALS/METHODS Four mini pigs underwent magnetic resonance (MR) evaluation of heart function and invasive electrophysiology assessment using electroanatomical mapping (EAM) before and 3 months after SBRT with single-fraction doses of 25Gy at the CTI. SBRT simulation, planning, and treatments were performed with the use of standard techniques. MR examination, including diffusion-weighted MRI, dynamic Gd-enhanced MRI, MR spectroscopy, and T2-weighted MRI were performed before and 3 months after SBRT, followed by histopathologic examination. One radiation oncologist and one radiologist evaluated the detectability and image quality for the irradiated site. MRI findings of the CTI radioablation lesion was compared to electrophysiology and histopathology findings. RESULTS A total dose of 25 Gy was delivered to the CTI (median volume of 25 cc) in a single procedure according to the radiation therapy oncology group. The mean radiation dose to the heart was 7 Gy. The maximal doses (< 0.05 cc) to the left and right coronary arteries were 14 Gy and 13 Gy, respectively. EAM visualized the irradiated site and confirmed clockwise conduction block across the CTI. Gd-enhanced T1-weighted imaging could only detect the irradiated site. There were no statistical differences in the image quality of the all MR images. CONCLUSION These data demonstrated the feasibility of SBRT for creating conduction block across the CTI in mini pigs. Gd-enhanced T1-weighted imaging was essential in assessing the radioablation lesion.

On the basis of epidemiological data and medical costs for patients with neuroblastoma, we have calculated the cost of mass screening for neuroblastoma with high performance liquid chromatography (HPLC) compared to the cost when it is not performed. If the sensitivity of the mass screening is 80 % and 22 000 infants are screened annually the cost will be 27809000 yen ($191800). If mass screening is not performed, the cost will be 28 446 000 yen ($196 200). The difference in cost (637 000 yen or $4 400) is fairly small. If the sensitivity is 75 % and 16 500 infants are screened, the difference is also small (174000 yen or $1 200). Therefore, mass screening with the HPLC method will not be an undue financial burden. But re-screening at an older age will be done with less financially favorable results, considering that the sensitivity may not be as high as that of the first screening and that mothers are somewhat reluctant about re-screening. The balance of the cost of mass screening by qualitative methods may also be less favorable, since the detection rate is low.

We corrected malpositioned continuous ambulatory peritoneal dialysis catheters in six patients using a new technique named the “push-pull method”. A gastric biopsy forceps was advanced through the catheter to near its tip. After manipulating the tip of the forceps through the abdominal wall, the forceps was opened and pulled out slowly. Repeated insertion and removal of the forceps induced the catheter to return to the pelvic cavity. This push-pull method was successful for Swan neck straight (n=2) and coiled (n=4) catheters in all patients. The time required for the procedure was only 5–10 minutes and there were no complications.

This paper is concerned with cavitation and erosion in submerged water jet. Impulse force generated by cavitation bubbles in the jet is measured by an impulse force gauge. The accumulated count of impulse force per unit time is shown against the impulse force and the standoff distance from the nozzle exit. It is found that there are maxima in impulse force at two different standoff distances of 30 and 80, which are the values normalized by the nozzle diameter (=1. 5 mm), and that the impulse force mainly appears in frequencies of a few Hz. Laser-microscopic observations show that erosion patterns on aluminum surfaces are classified into two ones.

The water-absorption properties of various vehicles were evaluated in rats whose stratum corneum had been stripped off over the application site. Polyethylene glycol (PEG) applied to the site effectively absorbed the exudate, but other applied ointments absorbed little exudate or water. By the PEG ointment, the exudate was absorbed continuously and extensively, and a linear relationship was obtained between the amount of water absorbed and the square root of time. To establish an acceptable alternative in vitro method for the design of a suitable ointment for treating skin ulcers with exudate, in vitro evaluations were performed by three methods: the Franz cell method, the cellulose membrane diffusion method and the spontaneous water uptake method. The membrane diffusion method, and the spontaneous water uptake method appeared to be acceptable alternatives to the in vivo method in evaluating the water absorption of ointments. It is a simple and useful method for preliminary formulation studies of ointments for the treatment of skin ulcers with exudate.

A large number of endocrine-disrupting chemicals(EDCs) show an estrogen-like action on the genital organs. Estrogen causes two kinds of irreversible changes in the reproductive system when given in fetal and early-postnatal period in mammals: change in the hypothalamo-hypophysio-gonadal system and change in the genital organ per se resulting in tumorigenesis. Natural and synthetic estrogens including some EDCs also give rise to permanent changes in various physiologically important systems in rodents exposed to them at developing period. The research trace reaching the finding of EDCs-phenomena indicates a successful approach to the achievement produced by the consistent, unfashionable and fundamental studies carried out for a long period of time.

We corrected malpositioned continuous ambulatory peritoneal dialysis catheters in six patients using a new technique named the "push-pull method". A gastric biopsy forceps was advanced through the catheter to near its tip. After manipulating the tip of the forceps through the abdominal wall, the forceps was opened and pulled out slowly. Repeated insertion and removal of the forceps induced the catheter to return to the pelvic cavity. This push-pull method was successful for Swan neck straight (n=2) and coiled (n=4) catheters in all patients. The time required for the procedure was only 5-10 minutes and there were no complications.

Transient elevation of serum free T4 (gestational transient hyperthyroxinaemia; GTH) occurs occasionally during normal pregnancy, especially in early gestation. However, the frequency of GTH and its clinical features remain unclear to date. The aim of this study was to determine the occurrence rate of GTH and the relation between serum levels of hCG, free T4 (fT4), and TSH in a large number of pregnant women.The four criteria of GTH were as follows: (1) no past history of thyroid disease, (2) negative tests for MCHA and TGHA, (3) no multiple pregnancies or trophoblastic disease and (4) transient hyperthyroxinaemia at less than 16 weeks of gestation. Thyroid function and hCG levels in 23,163 pregnant women were evaluated by mass sreening. If individual fT4 levels were more than the upper limit, blood re-sampling and the clinical and laboratory analysis of thyroid function were performed to exclude women with thyroid disease. The concentrations of hCG, fT4, and TSH in women with GTH and normal pregnant controls (n = 218) were compared. Regression analysis was performed for the comparison between hCG, fT4, and TSH levels in women with GTH.Blood samples were obtained using dried blood spots. Blood levels of fT4 was measured by radioimmunoassay, TSH and hCG were measured by fluoroimmunoassay. Anti-microsome antibody (MCHA) and anti-thyroglobulin antibody (TGHA) were measured by indirect agglutination reaction.GTH was observed in 66 of 23,163 women. The overall occurrence rate of GTH was 0.285%. In 22 of the 66 GTH women, serum TSH was undetectable. Using regression analyses, the concentration of fT4 was correlated with hCG levels in women with GTH (P0.05, r = 0.269), whereas the concentration of TSH was not correlated with hCG or fT4 level. The concentrations (M +/- SD) of fT4, TSH, and hCG in women with GTH were 42.5 +/- 12.3 pmol/l, 0.20 +/- 0.31 mU/l and 190.2 +/- 98.8 x 10(3) IU/l, whereas those of controls were 14.6 +/- 3.8 pmol/l, 1.43 +/- 1.25 mU/l and 60.1 +/- 45.1 x 10(3) IU/l. The concentrations of fT4 and hCG were significantly (P0.0001) higher than those of normal controls, and TSH was significantly (P0.0001) lower than those of normal controls.The occurrence rate of gestational transient hyperthyroxinaemia was 0.285%, and could possibly be attributed to increased levels of circulating hCG. Based on the data obtained from a large number of pregnant women, we propose gestational transient hyperthyroxinaemia as a definite clinical entity.

A 51-year-old woman began haemodialysis for chronic renal failure in February 1981. Symptomatic anaemia required treatment with recombinant human erythropoietin (rHuEPO) in February 1990 (3000 IU, twice weekly, intravenously). She developed influenza-like symptoms and treatment was withdrawn. In June 1994 rHuEPO was resumed at a very low dose of 100 IU subcutaneously three times weekly, and was increased gradually to 500 IU, without inducing any side-effects. At this dose the haematocrit was maintained at 22.0 – 25.0% and the symptoms of anaemia improved. In patients like ours, with influenza-like symptoms caused by rHuEPO therapy, dose escalation starting from an ultra-low dose may be effective in avoiding side-effects.

To clarify the abnormalities of coagulation and fibrinolytic systems on predialysis patients with chronic renal failure, we measured indices of coagulation and fibrinolytic systems in 33 predialysis patients whose creatinine (Cr) levels were over 3.0 mg/dl. We termed twenty-four patients with chronic glomerulonephritis the "CGN group". We also termed nine patients wit diabetes mellitus the "DM group". We measured thrombin.antithrombin III complex (TAT), alpha 2-plasmin inhibitor plasmin complex (PIC), D-dimer, protein C, protein S, thrombomodulin (TM), vitronectin, tissue plasminogen activator.plasminogen activator inhibitor-1 complex (tPAI-C) in theses two groups. Furthermore, we measured the same indices after 6 months in the CGN group. As a result, the plasma levels of both TAT, PIC, TM/Cr ration in the DM group were significantly higher that those in the CGN group, changes in both protein S activities and plasma levels of tPAI-C were reduced significantly after 6 months. In conclusion, the abnormalities of coagulation and fibrinolytic systems in predialysis diabetic patients were stronger than those in predialysis patients with CGN. Furthermore, these abnormalities were worsened after 6 months in predialysis patients with chronic renal failure.

We tried to discriminate between cases of spontaneous regression and non-regression during mass screening for neuroblastoma, taking advantage of differences in respect to the urinary homoranillic acid/vanillyl mandelic acid (HVA/VMA) ratio and the original tumor site among true positive, false negative and natural occurrence cases. After classifying them into a total of six groups depending on the two factors, ratio:1, 1-2 oror = 2 and tumor site: adrenal or extra-adrenal in origin, we calculated the mathematical probability of a given true positive case being one of spontaneous regression. A tumor of extra-adrenal origin was likely to regress spontaneously, especially one with an HVA/VMA ratio1 oror = 2 (82.2-100%). A tumor of adrenal origin with an HVA/VMA ratio1 seemed unlikely to regress spontaneously (0-4%). The present method, employing simple preoperative information, would be useful in future for the selection of true positive cases which should be observed without treatment.

1) By March 1992, 166,959 infants were screened in Sapporo City, and 35 patients were found to have this tumor. Incidence of this tumor in our screening was one among 5,059 babies examined. 2) Eleven infants among the group with negative screening at 6 months developed neuroblastoma later. The patients developed tumors one to several years after mass screening, and 2/3 of these cases showed high urinary HVA compared to VMA. 3) We postulate that some of the adrenal steroids, possibly cortisol, may play a role for the malignant progression of this tumor.

In patients undergoing chronic hemodialysis, marked anemia may cause decreased immunological function. To improve this anemia, we investigated the effect on immunological indices of recombinant human erythropoietin (rHuEPO) administration in 24 hemodialysis patients (13 males and 11 females) with renal anemia complications. Their mean age was 54.9 +/- 14.8 years and the mean duration of dialysis was 100.5 +/- 54.9 months. The subjects were treated with rHuEPO for 12 months, which helped to maintain a hematocrit value elevated by 5% from the baseline. Cell-mediated immunity and humoral immunity were assessed prior to as well as throughout the treatment period. Of the total number of patients studied, the anemia of 16 improved while 8 did not show signs of sufficient improvement. The improved group showed an increase in in vitro lymphocytic response to phytohemagglutinin (PHA), CD4/CD8, CD16 and serum IgM levels, while the CD8 level decreased significantly. Improvement in the general physical condition with rHuEPO treatment appeared to be associated with the changes in immunological indices, but the precise mechanism remains obscure.

Neonatal treatment of female mice with a synthetic estrogen, diethylstilbestrol (DES), results in adenosis, ovary-independent epithelial proliferation and cornification, and downgrowths in the vagina. Protein synthesis was examined in the vagina of 45-day-old, ovariectomized C57BL/Tw mice which had been given 5 daily injections of 2 micrograms DES or the oil vehicle alone from the day of birth, and in those of age-matched, ovariectomized mice given 3 daily injections of 0.1 microgram DES from 42 days of age. [35S]Methionine-labeled proteins of the vagina were analyzed by two-dimensional polyacrylamide gel electrophoresis. Major changes were observed in 11 proteins of the vagina in both neonatally and postnatally DES-exposed groups. Six of the 11 proteins were increased in expression, but one of the remaining proteins was decreased. In the group of neonatally DES-exposed mice alone, expressions of 4 proteins (MW. 128, 90, 46 and 44 kDa) were markedly increased. These results indicate that neonatal exposure of mice to DES induced ovary-independent, persistent alteration in the protein synthesis of the vagina.

Postpubertal injections of a synthetic estrogen, diethylstilbestrol (-DES), and an anti-estrogen, tamoxifen (-Tx), stimulate proliferation of vaginal and uterine epithelial cells of ovariectomized (OVX) adult mice. In vaginae of two groups of OVX mice treated neonatally with DES (DES-) and oil vehicle alone (Oil-), postpubertal injections of DES altered all and 25 out of 37 protein expressions examined, except for keratin polypeptides, respectively. Twenty-one of the DES- and Tx-altered protein expressions showed the same behavior, suggesting that Tx acts as an estrogen agonist on these proteins. Both neonatally Oil- and DES-treated OVX mice given postpubertal injections of DES (Oil-DES and DES-DES) and Tx (Oil-Tx and DES-Tx) showed similar histological changes in the vagina. Epithelial proliferation and superficial cornification were observed in vaginae of five groups of Oil-DES, Oil-Tx, DES-DES, DES-Tx and DES-Oil (given oil postpubertally) OVX mice. In these groups, 13 proteins showed the same behavior, implying that these proteins are related to proliferation and cornification of mouse vagina. The histology of the uterus in DES-Oil OVX mice closely resembled that in Oil-Oil OVX mice; in protein expressions of the uterus, 5 proteins in DES-DES and 7 proteins in DES-Tx OVX mice were different from those in Oil-DES and Oil-Tx OVX mice, respectively. These findings indicate that neonatal DES treatment alters the behavior of the vagina and uterus in response to postpubertally administered DES and Tx.

Postpubertal progesterone injections slightly inhibited the proliferation and cornification of the vaginal epithelium induced by neonatal injections of diethylstilbestrol (DES). In vaginae of neonatally DES-exposed mice (DES mice), 9 protein expressions (PEX) appeared newly; 13 PEX decreased; 5 PEX increased compared to those in the controls. In vaginae of DES mice, PEX were altered by postpubertal injections of progesterone (DES-P); 3 PEX disappeared; 11 PEX were reversed to those in the controls. Progesterone injections impaired the proliferation of the vaginal epithelium, reversing 11 PEX compared to those in DES mice. In uteri of DES-P mice, 3 PEX increased, 2 PEX decreased, 2 PEX appeared and 1 PEX disappeared compared to those in DES mice. In conclusion, it was shown that postpubertal injections of progesterone alter the protein expression in the vagina and uterus of DES mice.

The distribution of the estrogen receptor (ER) was investigated immunohistochemically in the genital tract of 0 (day of birth)--to 50-day-old C57BL/Tw female mice given 5 daily injections of 2 micrograms diethylstilbestrol (DES) from the day of birth using an anti-ER monoclonal antibody. In the oviduct, cervix and vagina of control mice, both epithelial and stromal cell nuclei showed positive ER immunoreaction at 0 day. Nuclei of uterine stromal cells also showed strong ER immunostaining even at 0 day. In contrast, the ER in uterine epithelial cells began to appear by day 5. The number of positive epithelial cells and staining intensity gradually increased until 10 days. In neonatally DES-exposed mice, ER appeared in uterine epithelial cells 24 h after the injection. Polygonal cells appearing in the vaginal epithelium of DES-exposed 3-day-old mice showed weak ER staining. Neonatal DES exposure reduced staining intensity of columnar vaginal epithelial cells and uterine stromal cells at 3 days. Vaginal epithelial cells undergoing ovary-independent stratification and cornification in neonatally DES-exposed mice and normal estrous mice showed ER immunoreaction only in the basal cells.

Neonatal treatment of female mice with diethylstilbestrol (DES) results in genital tract abnormalities including ovary-independent vaginal proliferation and cornification. Protein profiles were examined in vagina and uterus from 45-day-old, ovariectomized C57BL/Tw mice which had been given 5 daily injections of 2 micrograms DES or oil vehicle alone from the day of birth, and in those from 45-day-old, ovariectomized mice given 3 daily injections of 0.1 microgram DES from 42 days of age. Proteins extracted were analyzed by two-dimensional polyacrylamide gel electrophoresis. In the vagina, expression of 37 non-keratin proteins was altered by postpubertal injections of DES as compared with the controls. In the neonatally DES-exposed vagina, expression of 26 of 37 proteins was altered as compared with controls. In the uterus, expression of 22 proteins was altered in the postpubertally DES-exposed group as compared with that in the control; however, the protein expression pattern of the neonatally DES-exposed group closely resembled that of the control except for one protein (no. 23, pI = 6.1, MW = 39.5 kDa) which was specifically increased in the neonatally DES-exposed group. By immunoblot analysis, 6 keratin polypeptides (49.5, 50, 52, 53, 57 and 58 kDa) were identified in vaginae of ovariectomized mice exposed neonatally and postpubertally to DES and of the controls. These results indicate that neonatal DES exposure induces organ specific alterations in the synthesis of proteins in mouse vagina and uterus.

In 35-day-old female ICR/JCL mice given 5 daily injections of 1 microgram diethylstilbestrol (DES) from the day of birth, a significantly higher incidence of polyovular follicles was found in the ovaries than in those of age-matched control mice. Gap junctions of granulosa cells of mature follicles in neonatally DES-exposed mice were larger than those of the controls. When stimulated by gonadotropins (PMSG and hCG) and caged with males, the number of tubal embryos in DES-exposed mice was less, but the rate of fertilization and development was not different compared to the controls. Division of oocytes collected from the ovaries of 40-day-old DES-exposed and control mice after stimulation of gonadotropins was examined 24 to 72 h after in vitro insemination to ascertain whether fertilization had occurred in oocytes from polyovular follicles. Seventy-seven % of oocytes from uniovular follicles of control mice developed up to 8-cell stage embryos following in vitro insemination; 66% of those from similar follicles of DES-exposed mice developed into the same stage. By contrast, only 47% of oocytes from polyovular follicles of DES-exposed mice showed the division up to 8-cell stage 72 h after insemination, indicating a significantly lower fertilization rate compared to the oocytes from uniovular follicles of control and DES-exposed mice. Without insemination, oocytes taken from the same pools in these experiments never divided during the period of manipulation and incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

In 35-day-old C57BL/Tw female mice given daily injections of 1 microgram diethylstilbestrol (DES) for 5 days from the day of birth, a significantly higher incidence of polyovular follicles (PF) were found in the ovaries than in those of age-matched control mice. Ovaries of prepubertal mice treated neonatally with oil or DES (DES mice) showed an enhancement of ovulation and luteinization following a combined treatment with eCG and hCG. Tubal ova in DES mice treated with eCG plus hCG were surrounded by many granulosa cells. Incidence of PF in control mice was not changed by eCG plus hCG treatment. In contrast, PF incidence in DES mice was reduced by prepubertal injections of eCG plus hCG. A high incidence of PF was also found in newborn mouse ovaries transplanted for 30 days into ovariectomized adult hosts given DES injections, but not in ovaries transplanted into intact or ovariectomized DES-untreated hosts. When neonatal ovaries were cultivated in a serum-free medium containing DES for 5 days and then transplanted into ovariectomized hosts, PF were formed in the grafts, but not in DES-unexposed grafts. Oocytes from PF in DES mice were found to have a smaller capacity for fertilization when examined in vitro. The present study also demonstrated that neonatal ovaries exposed to estrogen in vivo or in vitro (which produces PF in prepubertal hosts) are capable of responding to gonadotropins given later, resulting in a reduction of PF incidence, and that exogenous estrogen acts directly on neonatal ovaries to induce PF.

We describe a simple and rapid method for measuring catecholamine metabolites in urine collected on filter paper. By this method, "high-performance" liquid chromatography, urinary vanillylmandelic acid and homovanillic acid can be measured within 15 min after being eluted from filter paper treated with tartrate buffer. One hundred urine samples can be pretreated within 2 h and 100-120 samples analyzed at 18-min intervals overnight in two chromatographic systems. Moreover, urinary creatinine also can be measured rapidly because of the use of microtiter plates. We applied this method in a mass screening program for early detection of neuroblastoma in infants. Between April 1986 and March 1987, two patients with neuroblastoma were detected. In all, neuroblastoma has been detected in 17 of 88,887 infants so examined since April 1981.

Sixty-eight cases of neuroblastoma detected during mass screening being performed by 11 local self-governing bodies of Japan were studied concerning age, clinical stage, and survival rate. Their average ages at the first screening, at the beginning of thorough examination, and at the start of therapy were 215.7, 245.6, and 264.7 days, respectively. This screening, which is aimed at the 6-month-old infants is considered to be acceptable, on the grounds that the majority of the cases (95.6%) were asymptomatic at the time of thorough examination, that for most of the cases (98.5%) the initiation of therapy was at under 1 year of age, and that their 60-month survival rate was 87.5%. However, there seems to be room for discussion of the best age at which subjects should be screened, because the 68 cases included some patients already with advanced stages at the time of thorough examination, and because "false negative cases" were identified besides the 68 positive cases.

We describe a microassay for measuring galactose (Gal) and galactose 1-phosphate (Gal-1-P) in dried blood spots. After a coupled enzyme reaction involving galactose dehydrogenase (GADH, EC 1.1.1.48) and alkaline phosphatase (AP, EC 3.1.3.1) in a microplate well, NADH fluorescence is measured by a highly sensitive fluorometric microplate reader, capable of rapid measurement of fluorescence (2 min per 96 samples). Within- and between-run CVs for measurements of Gal at 90 mg/L with Gal-1-P at 130 mg/L were both less than 5% (n = 8), and analytical recoveries for Gal at 90 mg/L and Gal-1-P at 130 mg/L were 98% and 92%, respectively. Five hundred dried blood-spot samples can be assayed within 2 h, with full calculation of results by an on-line microcomputer. This rapid and reliable assay system is very useful for the routine screening of newborns for galactosemia.

Female C57BL/Tw mice were given 5 daily injection of 20 micrograms testosterone (T), 100 micrograms 4-hydroxy-4-androstene-3, 17-dione (4-HA), 100 micrograms 6-methylene-4-pregnene-3, 20-dione (6-MP), 4-HA + 6-MP, T + 4-HA, T + 6-MP, and T + 4-HA + 6-MP starting on the day of birth. The animals were ovariectomized at 30 days or 90 days of age and were killed at 150 days. The incidence of polyovular follicles (PF) at 30 days of age was significantly increased by neonatal treatment with T. By contrast, the PF incidence was significantly reduced by injections of 4-HA given simultaneously with T. Neonatally T- or T + 6-MP-treated 90-day-old mice had ovaries containing follicles and hypertrophied interstitial cells but no corpora lutea. By contrast, T + 4-HA (64%)- and T + 4-HA + 6-MP (82%)-treated mice had ovaries with corpora lutea. In the T + 4-HA + 6-MP (82%)-treated mice had ovaries with corpora lutea. In the T + 4-HA + 6-MP-treated, 150-day-old, ovariectomized mice, the number of mice showing vaginal epithelial stratification was significantly decreased as compared with T-treated mice. There were no significant differences in the number of layers, thickness, and mitotic rate of vaginal epithelium of T-treated mice compared with mice treated with T + 4-HA, T + 6-MP, or T + 4-HA + 6-MP. The present results indicate that development of PF and persistent anovulation are due to the direct action of estrogen (E) derived from T upon neonatal ovarian follicles and the neonatal hypothalamo-hypophysial system, and that T itself can induce ovary-independent vaginal changes, although 5 alpha-reduced androgen and estrogen derived from T seem to be more effective in this regard.

A mass screening system for the early detection by means of a vanillylmandelic acid test of neuroblastoma in 6-month-old infants in Japan has been developed in eight districts. 16 of the 281 939 infants screened by this test had neuroblastoma, equivalent to a very high incidence of 1 in 17 621. 15 of the 16 children with neuroblastoma are alive; the other child died 1 month after surgery. This mass screening system for neuroblastoma used in infancy can help to improve prognosis in infants with this malignant disorder.

Five to ten daily injections of 5--20 micrograms 17 beta-estradiol, 20--100 micrograms testosterone and its propionate, and 100 micrograms 5 alpha-dihydrotestosterone starting within 24 hours after bith caused estrogen-independent persistent proliferation and cornification of the vaginal epithelium in female mice of the C57BL/Tw, C57BL/Ms, A/Ms, and A/CrgI strains. The permanently changed vaginas frequently resulted in tumorigenesis. In C57BL/Tw mice, some undifferentiated cells, which survive only for a few postnatal days in normal animals, appeared to transform into large cells and formed nodules after neonatal administration of 17 beta-estradiol. The primary normal epithelium was replaced by a sheet of fused nodules that showed persistent proliferation and cornification independent of estrogen. Occurrence of the estrogen-independent vaginal epithelium in C57BL mice receiving neonatal injections of 20 micrograms estradiol for 5 days was prevented by injections of 200 IU VA when given simultaneously with the estradiol. Light cells were found by electron microscopy in the epithelium of estradiol-plus VA-treated mice. How these cells may have prevented the occurrence of the permanently changed vaginal epithelium is discussed.

Echocardiograms of two hundred and ninety normal fetuses at 18-39 weeks of gestation were studied with a real-time linear scanner (3.5 MHz). Aortic valvular diameter, pulmonary valvular diameter, mitral valvular diameter, tricuspid valvular diameter and total cardiac dimension were measured. These parameters increased with fetal growth. The ratios of pulmonary to aortic valvular diameter and of tricuspid to mitral valvular diameter were more than 1, suggesting right ventricular dominance in the fetal hearts. The cardio-thoracic ratio was about 50% in this period. The four-chamber view, left ventricular long-axis view, ventricular short-axis view, short-axis view of the great arteries, the view of the aortic arch and the view of the inferior and superior vena cava were also recorded. These six views could be obtained in 9.8% of all subjects at the same time. Detection rates for these views were the greatest in pregnancy at 22-33 (especially 28-29) weeks of gestation. The ventricular short-axis view and short-axis view of the great arteries were easily detected in the dorsoposterior position, whereas the view of the aortic arch was obtained in the dorsoanterior position. It was concluded that fetal echocardiography needed to be performed at least twice, at 22-23 and 28-29 weeks of gestation, and this would be useful in diagnosing some congenital heart diseases.

Bioavailabilities of three commercially available tablets and two trial tablets of nalidixic acid were determined. The correlation between in vivo bioavailability parameters and in vitro dissolution rates derived by eight methods (oscillating basket I and II, beaker I, II, and III, rotating basket, paddle and rotating flask) is discussed. Nalidixic acid and its metabolite, 7-hydroxynalidixic acid, were determined in serum by HPLC with a strong anion-exchange resin column. All parameters for nalidixic acid bioavailability showed significant differences among the tablets tested. The relationship between Cmax and AUC was not linear but concave. However, Cmax decreased linearly in proportion to the delay of Tmax. The rank order of tablets according to bioavailabilities was just the same as the rank order obtained when comparing dissolution rates determined by any of the above methods, with the exception of one tablet which had a poor disintegrating ability. The parameters for the rate of bioavailability (Cmax and Tmax) showed good linearity with 1/T50 determined by all of the methods. However, AUC showed significant correlation with neither T50 nor 1/T50.

Postcapillary or high-endothelial venules were preferentially distributed at the cortico-medullary junction of the thymus. In the lymph nodes these venules were located in he paracortical area between cortex and medulla. The venules in the thymus were surrounded by the so-called perivascular spaces, while those in the lymph nodes were not clearly surrounded by this space. The walls of the thymus venules were fenestrated. The reticular sheets surrounding the perivascular spaces were also fenestrated. These fenestrations facilitated the transmural passage of thymocytes through the vessels. In the lymph nodes where the venules had few fenestrations, lymphocytes in transmural passages were usually suspended between the high- endothelial cells. The macrophages were preferentially located in reticular meshes of the paracortical areas, and most of these cells were in contact with lymphocytes. This might indicate that the paracortical areas are the main interaction sites of lymph node lymphocytes and macrophages. Observations on the casted samples are also reported.

We describe a simple and rapid method for measuring catecholamine metabolites in urine collected on filter paper. By this method, "high-performance" liquid chromatography, urinary vanillylmandelic acid and homovanillic acid can be measured within 15 min after being eluted from filter paper treated with tartrate buffer. One hundred urine samples can be pretreated within 2 h and 100-120 samples analyzed at 18-min intervals overnight in two chromatographic systems. Moreover, urinary creatinine also can be measured rapidly because of the use of microtiter plates. We applied this method in a mass screening program for early detection of neuroblastoma in infants. Between April 1986 and March 1987, two patients with neuroblastoma were detected. In all, neuroblastoma has been detected in 17 of 88,887 infants so examined since April 1981.

An enzyme-linked immunosorbent assay (ELISA) for Cortisol in dried blood collected on filter paper has been developed. The ELISA method is very simple and rapid and has sensitivity, accuracy and precision. The detection limit of the ELISA is 10 ng/ml blood. The intra- and interassay coefficients of variation are 11.5-13.5% and 15.7-16.7, respectively. Using the ELISA for Cortisol and 17-hydroxyprogesterone (17-OHP) previously reported, dried blood from normal newborns including premature infants (n = 1583) and 21-hydroxylase deficiency (21-OHD) patient (n = 9) were analyzed. 17-OHP, Cortisol and 17-OHP/Cortisol ratio of normal newborns are 12.5 +/- 7.5 ng/ml (1.0-140 ng/ml), 89.0 +/- 64.3 ng/ml (10-1580 ng/ml) and 0.16 +/- 0.08 (0.01-0.86), respectively. In 21-OHD patients, 17-OHP is 109-1361 ng/ml, Cortisol is 35.1-146.7 ng/ml and 17-OHP/cortisol ratio is 1.84-12.2. It made recall rate less to measure Cortisol in addition to 17-OHP and to take 17-OHP/Cortisol ratio. Therefore, additional measurement of Cortisol to the primary 17-OHP screening test is advantaged in differentiating the normal newborns from the 21-OHD patients.

Further study on identification of metabolites of cinepazide, 4-(3,4,5-trimethoxycinamoyl)-1-(1-pyrrolidinyl)carbonylmethylpiperazine, in rats and men was carried out. New metabolites of cinepazide in rat urine were 4-[1-[4-(3,4,5-trimethoxycinnamoyl)-piperazinly]acetamidol-1-butanol (M-II), 4-(3,4,5-trimethoxycinnamoyl)-1-[1-(hydroxypyrrolidinyl)]carbonylmerthylpiperazine (M-IV) and 4-[1-[4-(3,4,5-trimethoxycinnamoyl)piperazinyl]acetamidol]-1-butyric acid (M-VI). Structures of two mono-O-demethylated metabolites of cinepazide predicted in the previous paper were assigned as 3-hydroxy-4,5-dimethoxycinnamoyl and 4-hydroxy-3,5-dimethoxycinnamoyl compounds (M-III and M-V) by the FT-NMR spectrometry measured in a mixture of CDC13-C6D6 (1:1, v/v) after methylation with dimethylsulfare-d6. The corresponding pyrrolidone metabolite of cinepazide was undetectable in rat urine. New metabolites of cinepazide in rat feces were found to be 4-(3,5-dihydroxy-phenethylcarbonyl)-1-(1-pyrrolidinyl)carbonylmethylpiperazine (M-VIII) and 4-(3,5-dihydroxycinnamoyl)-1-(1-pyrrolidinyl)carbonylmethylpiperazine (M-IX). Metabolites of cinepazide in human urine were studied by both methods--high performance liquid chromatography and mass spectrometry with ion cluster technique. Unchanged drug was mainly excreted in the urine, and M-VI was the major urinary metabolite. The existence of M-IV and M-V was suggested by high performance liquid chormatography. The corresponding pyrrolidone was also undetectable in human urine.

The bioavailability of nalidixic acid in beagles was determined using the same tablet formulations previously tested on humans and was compared with bioavailability in humans and with in vitro dissolution rates. The beagle bioavailability test provided lower power in all of the bioavailability parameters than did the human test. Tmax of the tablets did not greatly differ in beagles, although in humans a wide variation of Tmax was seen. A linearity between Cmax and AUC0-infinity was observed in beagles, but Tmax did not show a linearity with Cmax or AUC in dogs, which is quite different from the relations observed in humans. The rank order of tablets according to Cmax was exactly the same between humans and beagles. AUC also showed the same rank order between humans and beagles except for on tablet with a poor disintegrating ability. A significant correlation between human and beagle Cmax values was obtained (r = 0.8952; p less than 0.05), but not between human and beagle AUC and Tmax values.

An enzyme-linked immunosorbent assay (ELISA) for 17 alpha-hydroxyprogesterone (17-OHP) in dried blood collected on filter paper has been developed. The ELISA method is very easy and rapid and has sensitivity, accuracy and precision. The detection limit of the ELISA method is 0.5 ng/ml blood. Intra- and interassay coefficients of variation are 7.0 approximately 8.3% and 6.7 approximately 10.5%, respectively. Correlation between the ELISA method and previously reported enzyme immunoassay is good (r = 0.961). 17-OHP values of neonates with congenital adrenal hyperplasia (CAH) are extremely high compared with normal neonates (11.0 +/- 5.9 ng/ml blood). The present study demonstrates that the ELISA method for 17-OHP is applicable to neonatal screening for CAH.

From the Childhood Cancer Registry in Japan, incidence of neuroblastoma was 1 among 100,000-150,000 children under the age of 15 years, and 25% of them were under 1 year old. But, mass-screening for this tumor in infancy showed incidence of detection of 1:5,000 babies. Even if we presume that all of the tumor cases--if not screened with the risk of developing later--were detected in infancy, this is more than the expected incidence. To explain this, we postulated 4 patterns in the natural growth of this tumor. After the start of mass-screening, the number of the patients with this tumor in much earlier stage and younger age increased and this was the main cause of improvement of the results of treatment.

The growth of transplanted carcinomas originating in a gonadectomized C3H male mouse was much faster in intact male mice than in intact or gonadectomized females. Administration of 17beta-estradiol or testosterone propionate in pellets resulted in a marked acceleration of the carcinoma growth in intact female mice. Progesterone and deoxycorticosterone acetate exerted no influence on the carcinoma growth. Neither gonadotropins (pregnant mare's serum gonadotropin plus human chorionic gonadotropin) nor ACTH accelerated the growth. In contrast, cortisone acetate in pellets markedly reduced the growth rate of carcinoma transplants when administered simultaneously with the transplantation. Cortisone acetate also arrested the growth-accelerating effect of 17beta-estradiol on transplanted carcinomas.

Ovary-indepedent (estrogen-independent) irreversible proliferation and cornification on the vaginal epithelium in ovariectomized mice caused by neonatal injections of 20 microng estradiol-17 beta (E2) was suppressed by injections of 200 IU vitamin A acetate (VA) given simultaneously with E2. More than 2 months after the combined treatment were required for the suppression. A new type of cell of low electron density (Light cells) appeared at 40-120 days of age on the basal lamina of the vaginal epithelium in neonatally E2 plus VA-treated ovariectomized mice only at 50 days. Light cells contained small amounts of organelles. Columnar basal cells resembling those in normal estrous mice were also found in the E2 plus VA-treatedmice at 70-120 days. In one of the 120-day-old, E2 plus VA-treated mice, the epithelium consisted of only 2-3 layers of cells. Neither Light cells nor columnar cells were found in the atrophic epithelium.