Visceral adipose tissue measured by computed tomography and high-grade prostate cancer after radical prostatectomy.

We assessed the association between obesity measurements including visceral adipose tissue (VAT), measured by computed tomography, and the risk of high-grade prostate cancer after radical prostatectomy. We investigated 296 patients who were diagnosed with prostate cancer and underwent radical prostatectomy. Data extracted from medical records included age, body mass index (BMI), VAT, pretreatment prostate-specific antigen (PSA) levels and Gleason score (GS). We performed logistic regression to examine the association between indicators of obesity and a higher GS (⩾4+3). Among the 296 patients, 107 (36%) had a higher GS. After controlling for age and PSA, BMI was not associated with GS (odds ratio, OR=1.039, 95% confidence interval, CI=0.943-1.145; P=0.437). BMI had different effects on GS depending on VAT. When the data were stratified by the median VAT value, a higher BMI was significantly associated with a higher GS in patients with VAT⩾130.5 cm2 (OR=1.218, 95% CI=1.028-1.443; P=0.022), but not in those with VAT<130.5 cm2 (OR=0.912, 95% CI=0.783-1.062; P=0.236). A higher BMI was associated with an increased risk of high-grade cancer only in patients with more VAT.

Relationship between drug lag and factors associated with clinical trials in Japan.

WHAT IS KNOWN AND OBJECTIVE: Drug lag is a major public concern in Japan. During the development of new drugs, some factors related to clinical trials in the marketing application package, such as trial design and the number of trials, can affect drug approval. The aim of this study was to determine whether those clinical trial factors were associated with drug lag in Japan. METHODS: We investigated new drug applications for new molecular entities that were approved in Japan between April 2009 and March 2012. We collected information on clinical trials in the marketing application package from review reports. RESULTS AND DISCUSSION: We constructed a multiple regression model to predict drug lag using the review period, use of foreign clinical trial data, the number of confirmatory trials, the design of the pivotal trial, failures of confirmatory trials and the death rate (n = 59). No use of foreign trial data was significantly associated with a longer drug lag (84% increase; 95% confidence interval [CI], 1·03-3·29). Compared to the open-label, one-armed design, drugs that underwent pivotal trials of placebo-controlled superiority, active-controlled superiority and active-controlled non-inferiority designs had a significantly shorter drug lag (74% decrease, 95% CI: 0·08-0·83; 74% decrease, 95% CI: 0·07-0·99; and 85% decrease, 95% CI: 0·04-0·58, respectively). WHAT IS NEW AND CONCLUSION: Our findings suggest that new drug application packages that do not use data from foreign clinical trials and that involve pivotal trials of open-label, one-armed design contribute to drug lag in Japan. To reduce this lag, improved strategies for the development of new drugs should be identified.

Relationship between body mass index and infertility in healthy male Japanese workers: a pilot study.

Our aim was to evaluate the relationship between men's body mass index (BMI) and infertility and to examine the effects of factors related to metabolic syndrome such as hypertension, dyslipidaemia, and impaired glucose tolerance. Our sample comprised 74 healthy Japanese workers at a company who were married between 2003 and 2005. The outcome variable was whether a baby was born during the study period (median follow-up period, 20 months; range, 8-42 months). Data for BMI and other factors were obtained from the results of an annual health checkup in the year of each employee's marriage. Forty-seven men (64%) did not father a baby. Having a baby was significantly associated with a low BMI (21.4 versus 23.2 kg m(-2); P = 0.006). A Cox proportional hazard regression model was performed to assess the association of BMI with fathering a baby. Adjusting for age, systolic and diastolic blood pressure, low density lipoprotein cholesterol, triglycerides, and haemoglobin A(1C), higher BMI was significantly associated with not fathering a baby (hazard ratio, 0.80; 95% confidence interval, 0.67-0.95; P = 0012). High BMI in men was independently associated with an increased risk of not siring a child.

Blood lead and erythrocyte protoporphyrin levels in association with smoking and personal hygienic behaviour among lead exposed workers.

AIMS: To investigate the effects of smoking and personal hygienic behaviour on blood lead (BPb) and free erythrocyte protoporphyrin levels (FEP) in lead exposed workers. METHODS: Subjects were 105 lead exposed male workers in a battery recycling plant during the years 2000-03. BPb and FEP were measured as part of the ongoing occupational surveillance. Each worker completed a questionnaire for assessment of smoking and four measures of personal hygienic behaviour (glove and mask use, hand and face washing before meals during working hours). RESULTS: Statistically significant decreases in mean BPb and FEP occurred during the three years. The proportion of BPb reduction in the non-smoking workers was significantly higher (mean 24.3%) than in the smoking workers (15.3%). When the workers were classified into three groups (excellent, good, and poor) based on the four personal hygienic behavioural indicators, the greatest decreases of BPb and FEP were observed in the non-smoking workers of the excellent group. CONCLUSIONS: The consistent use of protection devices and cleanliness at work appeared to contribute to the lowering of BPb and FEP. Cessation of smoking in the workplace was also of importance.

Relationship between shunt-dependent hydrocephalus after subarachnoid haemorrhage and duration of cerebrospinal fluid drainage.

Subarachnoid haemorrhage (SAH) patients in Fisher group 3 have a high risk of vasospasm and chronic hydrocephalus. We have provided cisternal irrigation combined with a head-shaking method for preventing vasospasm in SAH patients. We investigated 76 SAH patients in Fisher group 3 who received cisternal irrigation with head-shaking to evaluate the relationship between the occurrence of hydrocephalus and various clinical factors, including duration of cerebrospinal fluid (CSF) drainage. Chronic hydrocephalus occurred in 25 patients (33%). The occurrence of hydrocephalus was associated with longer duration of CSF drainage (median, 13 days versus 9 days). By logistic regression analysis using significant factors, including age, preoperative neurological grade and Glasgow Outcome Scale, only the duration of drainage was independently associated with the occurrence of hydrocephalus (Odds ratio = 1.18 per day; 95% confidence interval, 1.02- 1.36). These results indicate that long duration of CSF drainage for preventing vasospasm may increase the occurrence of hydrocephalus.

Discovery of a novel CCR3 selective antagonist.

In searching for a novel CCR3 receptor antagonist, we designed a library that included a variety of carboxamide derivatives based on the structure of our potent antagonists for human CCR1 and CCR3 receptors, and screened the new compounds for inhibitory activity against 125I-Eotaxin binding to human CCR3 receptors expressed in CHO cells. Among them, two 2-(benzothiazolethio)acetamide derivatives (1a and 2a) showed binding affinities with IC50 values of 750 and 1000 nM, respectively, for human CCR3 receptors. These compounds (1a and 2a) also possessed weak binding affinities for human CCR1 receptors. We selected la as a lead compound for derivatization to improve in vitro potency and selectivity for CCR3 over CCRI receptors. Derivatization of la by incorporating substituents into each benzene ring of the benzothiazole and piperidine side chain resulted in the discovery of a compound (1b) exhibiting 820-fold selectivity for CCR3 receptors (IC50 = 2.3 nM) over CCR1 receptors (IC50 = 1900 nM). This compound (1b) also showed potent functional antagonist activity for inhibiting Eotaxin (IC50 = 27 nM)- or RANTES (IC50 = 13 nM)-induced Ca2+ increases in eosinophils.

Design, synthesis, and discovery of a novel CCR1 antagonist.

The CC chemokines may play an important role in the pathogenesis of chronic inflammatory diseases including rheumatoid arthritis, and their effects are thought to be mediated through CCR1 receptors. Several nonpeptide CCR1 receptor antagonists that showed high affinity for human CCR1 receptors have been identified; however, their effectiveness in animal models of inflammatory diseases has been scarcely demonstrated, probably due to species selectivity of the antagonists. To elucidate the pathophysiological role of CCR1 receptors in murine models of disease, we looked for a potent antagonist for both murine and human CCR1 receptors. Screening of our chemical collection for inhibition of (125)I-MIP-1alpha binding to human CCR1 receptors transfected in CHO cells led to the identification of xanthene-9-carboxamide 1a as the lead compound. Derivatization of 1a by quaternarizing the piperidine nitrogen with various alkyl groups and by installing substituents into the xanthene moiety dramatically improved the inhibitory activity against both human and murine CCR1 receptors. As a result, 2q-1 showing IC(50) values of 0.9 and 5.8 nM for human and murine CCR1 receptors, respectively, was discovered. This compound is the first murine CCR1 receptor antagonist and may be a useful tool for clarifying the role of CCR1 receptors in murine models of disease.

Identification of a potent and nonpeptidyl ccr3 antagonist.

CCR3 is expressed in a variety of leukocyte subsets, especially eosinophils, and may be involved in allergic disorders such as atopic asthma. To clarify the pathophysiological roles of CCR3 in allergic disorders, we developed a nonpeptidyl CCR3 antagonist. This antagonist, which is referred to as "Compound X," that inhibited the binding of [(125)I]Eotaxin to CHO cells transfected with human CCR3 with an IC(50) value of 2.3 nM. In human eosinophils, Compound X also inhibited Eotaxin-induced increases in intracellular Ca(2+) concentrations and chemotaxis. Thus, Compound X appears to be a highly potent CCR3 antagonist. These findings suggest that Compound X may be a useful tool for elucidating the pathophysiological roles of CCR3 in a variety of allergic disorders.

[Rapidly enlarging supratentorial ependymoma in a child presenting initially with a small calcified lesion: case report].

The authors report an unusual case of a 11-year-old boy whose supratentorial ependymoma showed rapid growth. He had had generalized convulsive seizures when he was 9 years old. On an initial CT scan a small calcified lesion was identified adjacent to the right sensorimotor cortex. Repeated CT scans showed no interval change in the size of the tumor for 16 months. Then, he suffered an acute onset of left hemiparesis. The neuroimaging studies demonstrated a huge tumor with a large cyst in the right parietal region which had not been observed on CT scan 7 months before. Total removal of the tumor was performed and the histopathological diagnosis was ependymoma with no evidence of malignancy. However, MIB-1 staining of the specimen revealed a high index of 1 proliferative potential up to 25% in some area. The high score of MIB-1 staining correlated well with the rapid clinical course of this histologically benign ependymoma. The small calcified lesion demonstrated on the initial CT scan in this case is considered to have been a low grade ependymoma and to have abruptly transformed into a higher grade, one resulting in rapid enlargement. The authors stress that small intracranial calcified lesions should be carefully followed up by repeated neuroimaging studies at short intervals.

Coexpression studies with endothelin receptor subtypes indicate the existence of intracellular cross-talk between ET(A) and ET(B) receptors.

Human Girardi heart cells expressing endothelin ET(B) receptors (GH(B) cells) were transfected with human ET(A) cDNA, and coexpression of ET(A) and ET(B) in the ratio of 4:6 was demonstrated by Scatchard analysis. [125I]Endothelin (ET)-1 binding to ET(A)-transfected GH cells (GH(AB) cells) was displaced by an ET(A) antagonist, BQ-123, in a biphasic manner. An ET(B) agonist, BQ-3020, and an ET(B) antagonist, BQ-788, inhibited [125I]ET-1 binding to GH(AB) cells in a monophasic manner with low affinities (IC50 = 2,800 and 890 nM, respectively); IC50 values for ET(B) receptors seemed to be as weak as those for ET(A) receptors. However, BQ-3020 and BQ-788 had a high affinity for ET(B) receptors in a binding experiment using [125I]ET-1 in the presence of 1 microM BQ-123, where ET(A) receptors are masked (IC50 = 0.49 and 0.89 nM, respectively). The ET(B)-mediated increase in intracellular calcium concentrations in GH(AB) cells was not affected by 0.1 microM BQ-788 alone but was inhibited significantly by the same concentration of BQ-788 in combination with 10 microM BQ-123. ET-1 suppressed forskolin-stimulated accumulation of cAMP through the activation of ET(A) and ET(B) in GH(AB) cells; 1 microM BQ-123 or BQ-788 inhibited the suppression by only 20%, whereas a mixture of BQ-123 and BQ-788 (1 microM each) completely inhibited the cAMP decrease. These findings suggest that the stimulation of ET(A) receptors with ET-1 results in a lowering of the affinity of BQ-3020 and BQ-788 for ET(B) receptors in GH(AB) cells. We conclude that there is intracellular cross-talk between ET(A) and ET(B) receptors in GH(AB) cells.

ETB-mediated regulation of extracellular levels of endothelin-1 in cultured human endothelial cells.

Our previous report suggests the important role that ETB receptors play in the clearance of circulating endothelin (ET)-1. The present study confirmed this finding by measuring the extracellular levels of immunoreactive (ir) ET-1 and intracellular contents of prepro (pp) ET-1 mRNA in the presence and absence of ETA- and ETB-selective antagonists (i.e., BQ-123 and BQ-788, respectively) in cultured human umbilical vein endothelial cells (HUVECs). ET-1 was secreted into the culture medium of HUVECs in a time-dependent manner with a plateau reached after incubation for more than 36 hr. In the presence of 10 microM BQ-788, the irET-1 level was enhanced significantly (i.e., up to 180% of control at 48 hr) whereas BQ-123 had no such effect. Specific binding of [125I]ET-1 to HUVECs was inhibited strongly by BQ-788 (IC50 = 2.4 nM) but very weakly by BQ-123 (IC50 = 1.4 microM), indicating that BQ-788 has a potent affinity for ETB receptors in HUVECs. The expression of ppET-1 mRNA was not changed by BQ-788. Extracellular levels of ET-1 decreased gradually after cellular treatment with cycloheximide. This decrease was significantly inhibited by BQ-788 but not by BQ-123 and was non-existent when the cells were incubated at 4 degrees C (where internalization of the receptor protein is not likely). In conclusion, HUVECs secrete ET-1 which, in turn, is internalized after binding to ETB receptors on HUVECs.

[3H]BQ-123, a highly specific and reversible radioligand for the endothelin ETA receptor subtype.

The mode of binding of [3H]BQ-123 (cyclo(-D-Trp-D-Asp-[prolyl-3,4 (n)-[3H]]Pro-D-Val-Leu)), an endothelin receptor antagonist radioligand, was evaluated in the human neuroblastoma cell line SK-N-MC at 37 degrees C. Scatchard analysis indicated the presence of a single class of [3H]BQ-123 binding sites with a high affinity of 3.2 nM. [3H]BQ-123 binding achieved steady state within 7 min and dissociated with a half-time of 1.4 min, while [125I] endothelin-1 binding barely reached a steady state even after 6 h and showed little dissociation. [3H]BQ-123 binding was sensitive to endothelin-1 and endothelin-2 (Ki values = 0.058 and 0.10 nM, respectively) and the endothelin ETA receptor-selective antagonist BQ-123 (Ki = 3.3 nM), while showing low affinity for endothelin-3 (Ki = 50 nM), the endothelin ETB receptor-selective agonist BQ-3020 (Ki = 970 nM) and other bioactive peptides. Thus, [3H]BQ-123 is a specific and reversible radioligand for endothelin ETA receptors. The rapid reversibility of [3H]BQ-123 binding should provide a tool for estimating the equilibrium inhibition constants (Ki values) of various compounds for endothelin ETA receptors.

Collagenase activity and tissue inhibitor of metalloproteinases-1 (TIMP-1) content in human whole saliva from clinically healthy and periodontally diseased subjects.

Total TIMP-1 concentration in whole saliva of periodontally diseased subjects, 137 +/- 67 ng/ml (mean +/- SD), was clearly lower (p < 0.001) than that of clinically healthy subjects, 273 +/- 145, and that of edentulous subjects, 332 +/- 121. On the contrary, both active [1.58 +/- 0.35 units/ml (mean +/- SD)] and total (2.08 +/- 0.25) collagenase activities in TIMP-1-free whole saliva of diseased subjects were significantly higher than the activities (0.14 +/- 0.14 and 0.50 +/- 0.27, respectively) in TIMP-1-free whole saliva of healthy subjects. Most of the total collagenase in whole saliva of healthy subjects consisted of procollagenase, while mainly active collagenase was present in whole saliva from patients with periodontal diseases. Significant reciprocal changes of TIMP-1 and collagenase levels, that is, increase in TIMP-1 concentration and decrease in collagenase activity, were observed after the initial therapy of periodontitis patients.

Antibody against a peptide corresponding to the extracellular domain of the luteinizing hormone/chorionic gonadotropin receptor stimulates testosterone production in rat Leydig cells.

In order to study the function of LH/CG receptor, we prepared four synthetic peptides (RP1, 2, 3, and 4 for residues 242-255, 49-66, 493-504, and 573-584) corresponding to extracellular domains of rat ovary LH/CG receptor and raised antibodies against RP1 and RP2 (anti-receptor peptide IgGs). These peptides and IgGs were assayed for inhibition of 125I-labeled hCG binding to Leydig cell membrane receptors. However, neither the peptides nor the IgGs inhibited hCG binding to the receptor at doses up to 10(-3) and 10(-5) M, respectively. On the other hand, although anti-receptor peptide IgGs did not show inhibition of hCG binding to the receptor, it was found that one of the IgGs (anti-RP1IgG) was bound to the cell membrane and stimulated testosterone production in rat Leydig cells. F(ab')2 fragment lacking the N-linked sugar chain in the IgG molecule, whose sugar chains are similar to those of hCG, was prepared from anti-RP1IgG. Anti-RP1F(ab')2 was still bound to the cell membrane but no longer stimulated testosterone production. These results suggested that when LH/CG receptor binds to a glycoprotein, even if it is different from the native ligand hCG, it may interact with sugar chains of the glycoprotein to cause signal transduction. Thus, a lectin-like domain in or near the receptor may play a key role in the receptor function.

[Dual nuclides SPECT for ventilation and perfusion study].

Dual nuclides SPECT using 81mKr and 99mTc-MAA for ventilation and perfusion study was performed in 24 subjects. Crosstalk of 81mKr to 99mTc-energy window was about 7.5% when ventilation and perfusion study were performed by 370 MBq of 81mKr gas and 185 MBq of 99mTc-MAA. Areas of low V/Q was significantly larger in SPECT study than in planar study, in 11 cases with various pulmonary diseases. High V/Q mismatches were also more clearly delineated in SPECT than in planar study. Dual nuclides SPECT study has advantages of obtaining V/Q distribution without movement artifacts and of simultaneous acquisition of ventilation and perfusion image. Area of high V/Q became larger in SPECT with crosstalk than in SPECT without crosstalk, but in the low V/Q area no significant difference was noted between SPECT with crosstalk and without crosstalk.

Steroidogenic activity of synthetic hybrid molecules composed of human chorionic gonadotropin and either the A or B chain of lectin ricin or horseradish peroxidase.

Human chorionic gonadotropin (hCG) is a glycoprotein consisting of noncovalently bound alpha- and beta-subunits which shows hormonal activity (stimulatory effect on testosterone production) toward rat Leydig cells. To modify the hormonal activity of hCG, hybrid molecules composed of hCG and other glycoproteins, either the A or B chain of lectin ricin (hCG-A and hCG-B) through disulfide bridges and horseradish peroxidase (hCG-HRP) through Schiff's base, were synthesized. Hormonal activity and the effect of these hybrids on [125I]hCG binding to rat Leydig cells were compared to those of native hCG. Modification of hCG resulted in a significant decrease in hormonal activity for hCG-HRP but not for hCG-A to approximately 1/100 and 1/10 that of native hCG, respectively. On the other hand, hCG-B unexpectedly showed hormonal activity similar to that of native hCG. These hybrids inhibited the binding of 125I-labeled hCG to rat Leydig cells with potencies of 1/10, 1/100 and 1/500 that of hCG for hCG-B, hCG-A and hCG-HRP, respectively. These results indicate that the B chain of ricin, the active component hCG-B, participated in stimulating testosterone production according to its own nature. Data which indicate that hybrids consisting of hCG subunits and the B chain of ricin (alpha-B and beta-B) stimulated testosterone significantly more than hybrids consisting of hCG subunits and A chain (alpha-A and beta-A) support the above finding.(ABSTRACT TRUNCATED AT 250 WORDS)

[High carboxylic acid level in the gingival crevicular fluid (GCF) of the patients with advanced periodontal disease].

The carboxylic acids are known to be produced by the bacteria in the dental plaque. The purpose of the present study is to investigate the concentrations of carboxylic acids in the whole saliva and the gingival crevicular fluid (GCF) of periodontal patients. The effects of these acids on human gingival fibroblasts were also examined. Nine males and 9 females patients were divided into two groups (Group A and Group B), according to the differences in probing depth (PD) and alveolar bone loss. Patients of Group A had gingivitis or slight adult periodontitis (PD less than or equal to 4 mm, alveolar bone loss less than or equal to 33%), whereas patients of Group B had moderate to advanced adult periodontitis (PD greater than 4 mm, alveolar bone loss greater than 33%). Carboxylic acids in the whole saliva and the GCF which were collected from the patients of Group A and Group B were examined using high performance liquid chromatography. The effects of two carboxylic acid salts (butyrate and formate) on the proliferation of human gingival fibroblasts from normal human gingiva were also examined by cell culture. Results obtained were as follows. 1. The concentrations of four carboxylic acids (acetic, propionic, succinic and butyric acids) in the GCF collected from patients of Group B were significantly higher than that of Group A. On the contrary, all carboxylic acid contents in the whole saliva were not significantly different between the patients of Group A and Group B. 2. Formate (1 or 3 mM) showed only a small degree of inhibition against the proliferation of human gingival fibroblasts, but butyrate (3 mM) almost completely (95.2%) inhibited the proliferation of human gingival fibroblasts. 3. The results of the present study may indicate that the carboxylic acids are involved in the initiation and development of the periodontal disease and that they can be one of the indicators in the diagnosis of the periodontal disease.

[A new treatment of malignant brain tumor -1: local injection of bleomycin (author's transl)].

This is a paper of new trial of treatment of malignant brain tumor by local injection of bleomycin (BLM). The new method of intraneoplastic BLM injection is as follows: in the cases ended up with subtotal or partial removal of the tumor, Ommaya's device was detained in the tumor bed, and its reservoir was fixed subcutaneous on the skull. 0.1 mg/kg to 0.2 mg/kg of BLM was injected by subcutaneous puncture into the reservoir every other day. Usual total dose of BLM was 30-80 mg. The patients were usually treated with 60Co-irradiation and immunotherapy after local injection of BLM. Twelve patients with malignant brain tumor were treated by the above mentioned new method and a follow-up study was done. One-year survival rate was 50% and three-year survival rate was 25%. Each case of primary sarcoma, medulloblastoma and malignant oligodendroglioma survived for a very long time. However, most of the patients especially those with glioblastoma died of recurrence in about one year or so inspite of temporary improvement of their clinical symptoms and clinical findings. In the autopsy cases of malignant gliomas, similar pathological findings were obtained around the tumor bed. In macroscopical view, the extensive necrotic foci and small haemorrhages were observed around the tumor bed not deeper than 2 to 3 cm from the surface of the cavity, and in the deeper part the tumor tissues were actively proliferating. Microscopically there was a severe coagulation necrosis of tumor cells with haemorrhage, collagenous tissue proliferation, fibrin deposit, increasing capillary vessels and infiltrating lymphocytes and granulocytes. Consequently, the scintillation scanning of BLM labelled 57Co was utilized to make clearance curves of the drugs in the patients with malignant brain tumor. The results were as follows: 57Co-BLM activity in the tumor tissue decreased about 70% 2-4 days after local injection of the drugs, whereas, it decreased about 70% 2-4 hours after intravenous injection of the drugs. From these results it could be presumed that locally injected BLM remained in the tumor tissues for a longer time and killed malignant tumor cells completely. However, an unfavorable fact was that locally injected BLM was retained only within the tumor tissue less than 2-3 cm apart from the cavity, showing no efficacy enough to prevent tumor proliferation in more remote area. In conclusion, the local injection of BLM seems to be very effective for the treatment of malignant brain tumors if it is used together with intravenous or intraarterial injection of other chemotherapeutic drugs.