A PTHrP Gradient Drives Mandibular Condylar Chondrogenesis via Runx2.

The mandibular condylar cartilage (MCC) is an essential component of the temporomandibular joint, which orchestrates the vertical growth of the mandibular ramus through endochondral ossification with distinctive modes of cell differentiation. Parathyroid hormone-related protein (PTHrP) is a master regulator of chondrogenesis; in the long bone epiphyseal growth plate, PTHrP expressed by resting zone chondrocytes promotes chondrocyte proliferation in the adjacent layer. However, how PTHrP regulates chondrogenesis in the MCC remains largely unclear. In this study, we used a Pthrp-mCherry knock-in reporter strain to map the localization of PTHrP+ cells in the MCC and define the function of PTHrP in the growing mandibular condyle. In the postnatal MCC of PthrpmCherry/+ mice, PTHrP-mCherry was specifically expressed by cells in the superficial layer immediately adjacent to RUNX2-expressing cells in the polymorphic layer. PTHrP ligands diffused across the polymorphic and chondrocyte layers where its cognate receptor PTH1R was abundantly expressed. We further analyzed the mandibular condyle of PthrpmCherry/mCherry mice lacking functional PTHrP protein (PTHrP-KO). At embryonic day (E) 18.5, the condylar process and MCC were significantly truncated in the PTHrP-KO mandible, which was associated with a significant reduction in cell proliferation across the polymorphic layer and a loss of SOX9+ cells in the chondrocyte layers. The PTHrP-KO MCC showed a transient increase in the number of Col10a1+ hypertrophic chondrocytes at E15.5, followed by a significant loss of these cells at E18.5, indicating that superficial layer-derived PTHrP prevents premature chondrocyte exhaustion in the MCC. The expression of Runx2, but not Sp7, was significantly reduced in the polymorphic layer of the PTHrP-KO MCC. Therefore, PTHrP released from cells in the superficial layer directly acts on cells in the polymorphic layer to promote proliferation of chondrocyte precursor cells and prevent their premature differentiation by maintaining Runx2 expression, revealing a unique PTHrP gradient-directed mechanism that regulates MCC chondrogenesis.

Expression of lacto series type 2 antigens in human renal cell carcinoma and its clinical significance.

We performed immunohistochemical examination of serial sections of human fetal and adult renal tissue as well as human renal carcinoma tissue, using monoclonal antibodies T5A7, 1B2, FH2, FH4, and FH6. These monoclonal antibodies were directed to lacto series type 2 antigens with sugar-chain structures: lactosylceramide, lactoneotetraosylceramide (paragloboside), Lex (a chemically well-defined fucosyl carbohydrate antigen), difucosyl Lex, and sialosyl-difucosyl Lex, respectively. The staining pattern in fetal renal tissue changed significantly according to the stage of organogenesis. In addition, expression of the antigens, especially paragloboside and sialosyl-difucosyl Lex, was closely related to the prognosis of the patient. These results suggest that the expression of a series of oncofetal antigens in development or differentiation of organs is reflected in the reversion to an immature pattern of antigenic expression in tumor tissue. The pattern of antigen expression in renal tumors offers a good criterion for ascertaining the degree of tumor differentiation and malignancy and is valuable for determining prognosis.

Disialosyl galactosylgloboside as an adhesion molecule expressed on renal cell carcinoma and its relationship to metastatic potential.

Aberrant glycosylation expressed in specific types of human cancer may define stage, direction, and fate of tumor progression. Well-studied examples are expression of sialosyl-Lewis(x) or sialosyl-Lewis(a) in colorectal carcinoma and histo-blood group A and H/Le(y) in lung cancer. In renal cell carcinoma (RCC), expression of sialosyl-Lewis(x) has no correlation with metastatic potential. Clinicopathological studies have revealed that the degree of expression of disialosyl galactosylgloboside (DSGG) and monosialosyl galactosylgloboside is correlated with metastatic potential (to lung and lymph nodes) of RCC and inversely correlated with patient survival. In the present study, we compared the adhesion of RCC lines to sections of various tissues measured by Stamper-Woodruff assay and other similar assays under dynamic flow conditions. Of the eight RCC lines tested, only TOS-1 (which expresses DSGG) bound strongly to lung tissue sections. TOS-1 did not bind to sections of liver, kidney, or lymph nodes. In the same eight RCC lines, we also compared expression of DSGG and monosialosyl galactosylgloboside (reflected by reactivity with RM1 and RM2), overall ganglioside patterns, and correlation with lung tissue-binding ability. Under both static and dynamic flow conditions, the binding of TOS-1 cells to lung alveolar tissue was correlated with their DSGG expression, i.e., the binding was inhibited by RM2 but not by RM1. This binding was also inhibited by sialidase but not by EDTA (i.e., it was CA 2+ independent). The other seven cell lines (TOS-2, TOS-M, SMKT-R1, -R2, -R3, and -R4, and ACHN), which do not express DSGG, showed much weaker adhesion to lung tissue. None of the eight cell lines showed E- or P-selectin-dependent adhesion. These results suggest the existence of a yet-uncharacterized sialoadhesive receptor++ that specifically recognizes DSGG. This receptor could be the binding target in RCC metastasis to lung.

Proliferative activity of intratumoral CD8(+) T-lymphocytes as a prognostic factor in human renal cell carcinoma: clinicopathologic demonstration of antitumor immunity.

Tumor-infiltrating lymphocytes, particularly CD8(+) T cells, could be a manifestation of antitumor immunity. We clinicopathologically analyzed the biological significance of tumor-infiltrating lymphocytes in 221 patients with renal cell carcinoma without preoperative treatments. More abundant infiltration of tumor tissue not only by CD8(+) but also CD4(+) T cells was associated with shorter survival of the patients, because of the positive correlation between the number of lymphocytes and representative tumor grade factors. This suggests that immune cell reactions are more pronounced as the tumor grade/biological malignancy progresses, probably because of increased antigenicity of tumor cells. We next analyzed the proliferative activity of CD8(+) T cells that infiltrated in tumor cell nests, which could also reflect antitumor immunity. Higher labeling index of Ki-67, a proliferation-associated antigen, among CD8(+) T cells in contact to tumor cells was associated with a longer survival by both uni- and multivariate analyses. Our data in human renal cell carcinoma suggest that infiltration of tumor tissue by T cells itself does not denote the efficacy of antitumor immunity because of its dependence on the biological malignancy of tumor cells, but infiltration of tumor tissue by CD8(+) T cells bearing more pronounced proliferative activity could reflect effective antitumor immunity. This concept would be important for future immunotherapy of human cancer.

The PEBP2beta/CBF beta-SMMHC chimeric protein is localized both in the cell membrane and nuclear subfractions of leukemic cells carrying chromosomal inversion 16.

The chromosomal inversion (16)(p13q22), which is associated with the M4-eosinophilia subtype of human acute myeloid leukemia, causes the fusion of two distinct genes. The polypeptide encoded by the chimeric gene, PEBP2p/CBFp-SMMHC, retains the ability to interact with, and dominantly interfere with the function of proteins possessing the Runt homology domain. The Runt protein homologs constitute the DNA binding subunit of the PEBP2/CBF transcription factor. We examined the subcellular localization of PEBP2beta/CBFbeta-SMMHC, as well as that of Runt protein homologs in leukemic cells carrying inversion 16 by immunoblot analysis. A significant amount of the PEBPbeta/CBFbeta-SMMHC protein was recovered from the nuclear fraction along with the Runt protein homologs. Furthermore, some of both polypeptides was retained in the DNA pellet that represents the material remaining after extraction of nuclear fraction with high salt. These observations suggest that the so-called dominant interfering effect of PEBPbeta/CBFbeta-SMMHC on PEBP2/CBF occurs inside the nucleus. In addition, we could detect PEBP2beta/CBFbeta-SMMHC in the cytoplasmic membrane fraction as well. The function of this membrane-located PEBP2beta/CBFbeta-SMMHC, if any, appears to be unrelated to that of Runt protein homologs.

Estrogen metabolism and impaired spermatogenesis in germ cell tumors of the testis.

We evaluated spermatogenesis in 36 patients with germ cell tumors [11 with seminoma (S) and 25 with nonseminoma (NS)] in terms of sperm concentration and histological score (Johnsen's mean score) of spermatogenesis in the ipsilateral and contralateral testes. We also measured the steroid concentration in the spermatic vein of the tumor-bearing side and performed biochemical and immunohistochemical studies of aromatase activity of the tumor to investigate the mechanism of exocrine and endocrine testicular dysfunction, with particular emphasis on the role of estrogen metabolism. The sperm concentration was significantly lower in patients with S (42.9 +/- 40.7 x 10(8)/mL) and NS (17.6 +/- 20.8 x 10(6)/mL) compared to normal adult men (114.4 +/- 41.2 x 10(6) mL; P < 0.01). The histological score was lower in patients with NS than in patients with S. The histological score was highest in the contralateral testis, followed by the ipsilateral testis far from the tumor and the ipsilateral testis near the tumor in both the S and NS groups. Serum levels of estradiol and hCG were significantly elevated in both the systemic and spermatic veins of patients with NS compared to normal values, but they were within normal limits in patients with S. The histological score count in the contralateral testis was significantly and inversely correlated with the tumor weight and serum levels of hCG and estradiol. Aromatase activity examined in 9 tumors (5 S and 4 NS) and 6 ipsilateral nonneoplastic testis (3 S and 3 NS) was significantly higher in both neoplastic and nonneoplastic testes in NS patients (tumor, 5.343 +/- 4.027; nontumor, 14.647 +/- 7.688 pmol/h.pg protein) compared to S patients (tumor, 0.622 +/- 0.408; nontumor, 1.979 +/- 1.164 pmol/h.pg protein). Aromatase immunoreactivity was observed in Leydig cells of the nonneoplastic testis in both S and NS patients and in interstitial or stromal cells in 16 of 25 of NS patients and none of S patients. Our results suggest that impaired spermatogenesis in patients with testicular germ cell tumor is caused by increased tumor size in both NS and S patients and/or by increased aromatization and in situ estrogen production in Leydig cells of the nonneoplastic testis and in interstitial or stromal cells of the tumor in patients with NS.

Qualitative difference of subcellular localization of tumor-associated carbohydrate (Le(x)) antigens in renal cell carcinoma and normal kidney.

Renal cell carcinomas are immunohistochemically positive for oligosaccharides with the Le(x) determinant (Gal beta 1----4[Fuc alpha 1----3]GlcNAc) and its derivatives, as oncofetal antigens, and their expression is closely related to a better prognosis of the patients. This study was designed to clarify the difference in antigen localization at the ultrastructural level between renal cell carcinoma and normal tissues. In normal kidneys, Le(x) detected by monoclonal antibody (MAb) FH 2 and sialylated extended Le(x) (sialyl Le(x)-i) by MAb FH 6 were identified along the plasma membrane of microvilli of proximal tubule epithelial cells, with occasional immunoreactivity along the basolateral plasma membranes. Intracellular localization was very sparse. Renal cell carcinoma showed localization of Le(x) and sialyl Le(x)-i antigens along the cell membrane and in the cytosol as aggregates or filaments. Immunoreactive materials were also observed in the lumen formed among carcinoma cells. The cytosolic immunoreactivity, not observed in the normal kidney, was regarded as "abnormal cytosolic accumulation" of the antigens. This pattern was more pronounced in clear-cell carcinoma. Pretreatment of specimens with chloroform-methanol, which extracts glycolipids, decreased immunoreactivity in carcinoma tissues, particularly that in the cytosol. The extracts contained substances immunoreactive for MAb FH6. Our study has demonstrated that (a) remarkable changes occur in the ultrastructural localization patterns of sialyl Le(x)-i and Le(x) in renal cell carcinoma and (b) considerable amounts of glycolipids are contained in the substances with sialyl Le(x)-i deposited in the cytosol of clear-cell carcinoma.

Association of renal cell carcinoma antigen, disialylgalactosylgloboside, with c-Src and Rho A in clustered domains at the surface membrane.

Disialylgalactosylgloboside (DSGG), defined by monoclonal antibody RM2, is a renal cell carcinoma (RCC)-associated antigen which mediates adhesion of RCC TOS-1 cells to certain lung tissue target cells. This adhesion process may initiate preferential lung metastasis of RCC. Ganglioside GM3 is a B16 melanoma-associated antigen which similarly adheres to target cells and promotes consequent metastasis. In view of the close association of GM3-enriched microdomain with transducer molecules c-Src, Rho A, and FAK in B16 cells, we investigated the organizational status of DSGG in RCC cell line TOS-1, with the following results: i) DSGG, but not monosialylgalactosylgloboside, showed extensive clustering at the TOS-1 cell surface; ii) a low-density membrane fraction isolated from TOS-1 cells contained >95% of cellular DSGG, although protein content in this fraction was <1% of total cellular protein; iii) this fraction contained c-Src, Rho A, and FAK, but not H-Ras; iv) c-Src and Rho A were co-immunoprecipitated with DSGG through anti-DSGG mAb RM2 (IgM) affixed to a column. These observations indicate that DSGG is clustered in RCC, as typified by TOS-1 cells, to form a microdomain in which it is closely associated with c-Src, Rho A, and FAK, and may constitute a functional unit as has been observed for GM3 with transducer molecules in B16 cells. The functional organization of such units may be essential in determining malignant properties of RCC cells.

G(M3) inhibits murine MET-2 tumor invasion and growth.

G(M3) has some important roles in cell-to-cell interaction and has proved to have an optimal concentration for fibronectin mediated cell adhesion. G(M3) content in murine bladder tumor (MBT-2) assessed by thin-layer chromatography was similar to human invasive bladder tumor. From glycolipid composition also, MBT-2 is considered as an appropriate model for human invasive bladder tumor. Anti-tumor effect of locally administered G(M3) On MBT-2 tumor was investigated. MBT-2 cells were injected subcutaneously into the right hind limb of CH3/HeSlc female mice on day 1. Tumor bearing mice were randomly placed on day 8 into G(M3) treatment, G(D3) treament, sialic acid treament and control groups. G(M3) was administered between tumor and fascia at 10 mu g in 0.1 ml, 1 mu g in 0.1 ml from day 8 to day 20 every other day, 7 times in total. Control group was given 0.1 mi of saline. G(D3) group was given 12.5 mu g of G(D3), and sialic acid group 2.5 mu g of sialic acid. The relative growth rates of control group, G(M3) 1 mu g group, G(M3) 10 mu g group on day 22 were 139+/-74, 56+/-39, 22+/-14, respectively, and statistically significant among these three groups (Mann-Whitney's U test p<0.01). There were no significant difference between control and G(D3) or sialic acid group. All of the 15 control mice had muscle invasion, however, of the 19 G(M3) 10 mu g administered mice, only 4 had muscle invasion. The incidence of muscle invasion between these 2 groups was statistically significant in chi(2) test (p<0.001). Locally administered G(M3) inhibited both invasion and growth of MBT-2 tumor. This mechanism could be explained by an important role of G(M3) in cell adhesion mediated by integrin and fibronectin interaction. These results may be applied to antiadhesion therapy of human invasive bladder tumor.

Effects of 5,7-dihydroxytryptamine and 6-hydroxydopamine on head-twitch response induced by serotonin, p-chloroamphetamine, and tryptamine in mice.

Head-twitch response (HTR) in mice was induced by intracerebroventricular injection of tryptamine (TRA) as well as serotonin (5-HT) and p-chloroamphetamine (PCA). Pretreatment with 5,7-dihydroxytryptamine enhanced both the 5-HT-induced and the TRA-induced HTR. The PCA-induced HTR, however, was attenuated by the drug. On the other hand, pretreatment with 6-hydroxydopamine did not alter the 5-HT response but enhanced both the PCA- and the TRA-induced response. These results suggest that 5-HT may directly stimulate the post-synaptic receptors, while the PCA response may be based on the release of endogenous 5-HT. The presynaptic component of the central serotonergic system does not appear to be involved in the TRA response. Both PCA and TRA may affect catecholaminergic systems which can suppress the response.

Head-twitch response induced by tyramine.

The intracerebroventricular (IC) administration of tyramine (TyA) induced a characteristic head-twitch response in mice pretreated with safrazine, a monoamine oxidase inhibitor. Safrazine-pretreated mice exhibited similar head-twitches following the IC treatment of serotonin (5-HT). The maximum dose of 5-HT which did not elicit head-twitches significantly potentiated TyA-induced head-twitches. Antiserotonergic drugs such as morphine and dimethothiazine antagonized TyA-induced head-twitches. A serotonergic denervator, 5,6-dihydroxytryptamine, potentiated head-twitch induced by TyA or 5-HT. Both TyA-induced and 5-HT-induced head-twitches were inhibited by dopamine and noradrenaline, while catecholaminergic denervators such as reserpine and 6-hydroxydopamine, and diethyldithiocarbamic acid, a dopamine-beta-hydroxylase inhibitor, increased the TyA response. These results indicate that head-twitches induced by TyA may be mediated via the serotonergic system and may inhibit the catecholaminergic system.

Identification of a 700-kb region of common allelic loss in chromosome bands 3p14.3-p21.1 in human renal cell carcinoma.

The short arm of chromosome 3 is considered to harbor one or more of the tumor suppressor genes taking part in the genesis of renal cell carcinoma (RCC). To define the localization of such putative tumor suppressor gene(s), we studied specific allelic loss on chromosome 3p by using 84 samples of RCC with nine microsatellite markers. We defined two commonly deleted regions in 3p14.3-p21.2: (1) region A, a 2-cM region between D3S1313 and D3S1592, and (2) region B, a 2-cM region between D3S1581 and D3S1289. The most frequent loss of heterozygosity was observed at D3S1067 (33 of 59, 55.9%), which is within region A. We further focused on region A and constructed a yeast artificial chromosome (YAC) contig and found that one YAC clone, which was 700-kb in size, harbored the entire region A. Using cosmid clones isolated from this contig, we also performed fluorescence in situ hybridization analysis and found that two of the tumors were homozygously deleted in this region. Our results strongly suggest the existence of a tumor suppressor gene in this region.

Congenital urethral and vesical diverticula allied to blind-ending ureters.

Blind-ending ureter is a well-recognized but rarely addressed abnormality that results in the urethral or bladder diverticulum. A girl and a boy are described with urethral diverticulum and bladder diverticulum, respectively, which are believed to be allied to blind-ending ureters.

Enzyme-linked immunosorbent assay detection of prostate-specific antigen messenger ribonucleic acid in prostate cancer.

OBJECTIVES: To develop a rapid, sensitive, reverse transcriptase-polymerase chain reaction (RT-PCR) prostate-specific antigen (PSA) messenger ribonucleic acid (mRNA) detection method by applying colorimetric enzyme-linked immunosorbent assay (ELISA). METHODS: Total RNA was extracted from 16 urogenital cancer cells (including PSA-producing LNCaP cells) from pelvic and inguinal lymph node aspiration biopsy samples from patients with prostate, bladder, and penile cancer, as well as from blood samples of 500 patients with urogenital cancer. We used rTth polymerase for RT and PCR. The RNA target was amplified by RT-PCR with dinitrophenyl-labeled primer. The PCR product was denatured and hybridized on a PSA-specific probe-coated microwell plate. RESULTS: In 1 6 cancer cell lines, only LNCaP cells expressed especially high PSA mRNA values, with an optical density (OD) greater than 3. In other cell lines, two testicular cells had relatively high ODs, 1.909 and 0.987, respectively. A high PSA mRNA value was obtained by fine needle aspiration from pelvic lymph node specimens of cytologically positive lymph nodes from patients with prostate cancer but not from patients with cytologically proved bladder or penile cancer. Sensitivity and specificity of fine needle aspiration samples were 70% and 100%, respectively. Blood tests obtained from patients with prostate cancer demonstrated high PSA mRNA values. CONCLUSIONS: The PSA mRNA RT-PCR ELISA method provides a sensitive photometric enzyme immunoassay for the detection of PSA mRNA, using nonradioactive techniques.

A possible mechanism of the tyramine-induced head-twitch response.

The effects of monoaminergic drugs on the head-twitch response (HTR) induced by intracerebroventricular (i.c.v.) injection of tyramine (TyA) in mice pretreated with safrazine, a monoamine oxidase inhibitor, were compared with effects on the response induced by the i.c.v. injection of serotonin (5-HT) in safrazine-pretreated mice. The HTR induced by both TyA and 5-HT were suppressed by i.p. injection of p-chlorophenylalanine. Chlorimipramine enhanced the 5-HT response but not the TyA response. Dimethothiazine, a serotonergic blocker, reduced both the TyA and 5-HT responses. The i.c.v. injection of p-chlorophenylalanine methylester resulted in a reduction of the TyA response but not of the 5-HT response. The i.c.v. injection of 5,6-dihydroxytryptamine (5,6 DHT) 1 day before the test suppressed the TyA response but enhanced the 5-HT response. The i.c.v. injection of noradrenaline reduced both the TyA and 5-HT responses. The i.c.v. injection of 6-hydroxydopamine, and i.p. injection of alpha-methyl-p-tyrosine and tolazoline accelerated the TyA response. These results suggest that the TyA response may be based on the release of endogenous 5-HT and can be suppressed by the catecholaminergic system.

Detection of prostate specific antigen messenger RNA from pelvic lymph node in prostate cancer.

We have adapted a sensitive method to detect pelvic lymph node metastasis using reverse transcriptase-polymerase chain reaction (RT-PCR) with primers specific for prostate-specific antigen (PSA) gene. Fine needle aspiration biopsy (FNAB) of pelvic lymph nodes was performed in 25 patients with prostate cancer. Each aspirated sample (0.05-0.1 mL) was divided into two parts: one for RNA extraction and RT-PCR to detect the fragment of PSA mRNA, and the other to smear on a slide glass for conventional cytology. RT-PCR detection of PSA gene was positive in 12 cases of FNAB samples, which included not only all 6 cytologically positive and 2 cytologically class III cases but also 4 of 17 cytologically negative cases. RT-PCR of FNAB samples of all 20 cases of bladder cancer were negative for the detection of PSA gene. RT-PCR for detection of PSA gene in FNAB samples may become a new diagnostic technique for detection of early lymph node metastasis in prostate cancer and an additional tool for cytologic diagnosis of prostate cancer.

Efficacy of an echo contrast agent, SH/TA-508, in color doppler sonography of mass lesions in urology.

We performed a clinical phase III study with a galactosebased ecoo contrast agent, SH/TA-508, to evaluate its efficacy, safety, and usefulness for mass lesions in urology. SH/TA-508 was prepared as a suspension containing stabilized micro-air bubbles by adding water for injection just before use. SH/TA-508 was administered into the antecubital vein at an initial dose of 300 mg/ml × 5 ml followed by higher doses of 400 mg/ml × 4 ml, 300 mg/ ml × 10 ml or 400 mg/ml × 8 ml when a sufficient effect was not obtained. Efficacy was evaluated by color Doppler signal enhancement, the duration of blood flow signal enhancement, and improvement of diagnostic capacity. Fifty-nine patients with mass lesions in the kidney, prostate, testis, adrenal gland, and bladder were enrolled in the study. Up to the third dose the cumulative efficacy rates (≥2+) of color Doppler signal enhancement and duration of blood flow signal enhancement were 92% and 87%, respectively. Consequently, diagnostic capacity in 76% of the patients was remarkably improved. A light transient angialgia occurred in one patient but no other clinically significant changes were observed. It was confirmed that SH/TA-508 is a safe echo contrast agent that offers satisfactory color Doppler signal enhancement in the urologic organs mentioned above.

Telomerase activity Simplification of assay and detection in bladder tumor and urinary exfoliated cells.

Detection of telomerase activity can differentiate malignant from benign cells. However, the original telomeric repeat amplification protocol (TRAP) methods had a number of limitations including a radioisotope labeling [α(32)P] dCTP [α(32)P] dGTP system. We developed digoxigenin labeled CX primer to detect telomerase activity without using radioisotope and attempted to detect telomerase activity of bladder tumor and exfoliated cells in bladder cancer patients. Telomerase activity was detected in 5 (71%) of 7 patients diagnosed with grade 1, 31 (97%) of 32 grade 2, and 11 (100%) of 11 grade 3 bladder tumors. In urinary exfoliated cells, 32 (82%) of 39 grades 1 or 2 bladder tumors were positive for telomerase activity but 20 (51%) of 39 were positive for urinary cytology (P < 0.01). Ten (91%) of 11 of grade 3 tumors were positive for telomerase activity and 11 (100%) of 11 were positive urinary cytology. Three of 100 noncancerous patients were positive for telomerase activity. Sensitivity, specificity, and positive predictive value of telomerase activity assay in urinary exfoliated cells were 84%, 97%, and 93%, respectively. Telomerase activity may be a useful diagnostic marker to detect the existence of immortal cancer cells in the urine.

Complications of laparoscopic and retroperitoneoscopic adrenalectomies in 370 cases in Japan: a multi-institutional study.

A total of 370 laparoscopic adrenalectomies, including 311 transperitoneal (TP) and 59 retroperitoneal (RP) approaches, were performed in nine urologic centers, where the laparoscopic adrenalectomy was first begun independently in Japan, and their affiliated hospitals between January 1992 and September 1996. The clinical diagnoses of those 370 adrenal diseases were primary aldosteronism in 155 patients, Cushing's syndrome in 61. preclinical Cushing's syndrome in 21. pheochromocytoma in 16, nonfunctioning adenoma in 87, complicated cyst in ten, myelolipoma in nine, adrenal cancer in four and other diagnoses in eight (table 1). There was no mortality in this series. Intraoperative complication rate was 33/370 (9%) in total: 26/311(8%) in the TP procedures and 7/59 (12%) in the RP procedures (table 11). Postoperative complication rate was 24/370 (6%) in total: 22/311 (7%) in the TP procedures and 2/59 (3%) in the RP ones (table 111). Conversion rates to open surgery in total, in the TP and in the RP procedures were 13/370 (3.5%), 10/311 (3.2%) and 3/59 (5.1 %). respectively (table IV). Although the RP procedure has a lower morbidity rate compared to the TP procedure, more skill is required to overcome the drawback of the narrow working space and fewer anatomical landmarks.

Effect of scopolamine on the electrical resistance of the paw pads of mice.

We examined the electrical resistance of the paw pads of mice under the same conditions as used previously in studies of the passive avoidance response. Administration of scopolamine (0.05-1 mg/kg, SC) 10 or 30 min prior to placement of animals in an experimental box resulted in a profound increase in electrical resistance. In contrast, subcutaneous injection of butylscopolamine (1-20 mg/kg), diazepam (1 or 2 mg/kg), or pentobarbital (10 or 20 mg/kg) did not substantially alter subject resistance. Scopolamine may act on the CNS to induce increased paw skin resistance.