Molecular basis of non-syndromic hypospadias: systematic mutation screening and genome-wide copy-number analysis of 62 patients.

STUDY QUESTION: What percentage of cases with non-syndromic hypospadias can be ascribed to mutations in known causative/candidate/susceptibility genes or submicroscopic copy-number variations (CNVs) in the genome? SUMMARY ANSWER: Monogenic and digenic mutations in known causative genes and cryptic CNVs account for >10% of cases with non-syndromic hypospadias. While known susceptibility polymorphisms appear to play a minor role in the development of this condition, further studies are required to validate this observation. WHAT IS KNOWN ALREADY: Fifteen causative, three candidate, and 14 susceptible genes, and a few submicroscopic CNVs have been implicated in non-syndromic hypospadias. STUDY DESIGN, SIZE, DURATION: Systematic mutation screening and genome-wide copy-number analysis of 62 patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study group consisted of 57 Japanese and five Vietnamese patients with non-syndromic hypospadias. Systematic mutation screening was performed for 25 known causative/candidate/susceptibility genes using a next-generation sequencer. Functional consequences of nucleotide alterations were assessed by in silico assays. The frequencies of polymorphisms in the patient group were compared with those in the male general population. CNVs were analyzed by array-based comparative genomic hybridization and characterized by fluorescence in situ hybridization. MAIN RESULTS AND THE ROLE OF CHANCE: Seven of 62 patients with anterior or posterior hypospadias carried putative pathogenic mutations, such as hemizygous mutations in AR, a heterozygous mutation in BNC2, and homozygous mutations in SRD5A2 and HSD3B2. Two of the seven patients had mutations in multiple genes. We did not find any rare polymorphisms that were abundant specifically in the patient group. One patient carried mosaic dicentric Y chromosome. LIMITATIONS, REASONS FOR CAUTION: The patient group consisted solely of Japanese and Vietnamese individuals and clinical and hormonal information of the patients remained rather fragmentary. In addition, mutation analysis focused on protein-altering substitutions. WIDER IMPLICATIONS OF THE FINDINGS: Our data provide evidence that pathogenic mutations can underlie both mild and severe hypospadias and that HSD3B2 mutations cause non-syndromic hypospadias as a sole clinical manifestation. Most importantly, this is the first report documenting possible oligogenicity of non-syndromic hypospadias. STUDY FUNDING/COMPETING INTERESTS: This study was funded by the Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology; by the Grant-in-Aid from the Japan Society for the Promotion of Science; by the Grants from the Ministry of Health, Labour and Welfare, from the National Center for Child Health and Development and from the Takeda Foundation. The authors have no competing interests to disclose. TRIAL REGISTRATION NUMBER: Not applicable.

Lower urinary tract function in spinal cord-injured rats: midthoracic contusion versus transection.

OBJECTIVES: To compare changes in lower urinary tract (LUT) function with modifications in pathways that regulate LUT function using two different animal models (incomplete and complete) of spinal cord injury (SCI). METHODS: Female Sprague-Dawley rats were used. SCI was made at Th8/9 by a contusion injury (contusion, n=9) or a complete transection (transection, n=9). Unoperated rats were used as normal controls (normal, n=6). LUT function was evaluated by micturition behavior in metabolic cages for 24 h and cystometry in awake animals. Immunocytochemical staining at the L6 spinal cord, spinal areas associated with LUT, was performed to identify descending modulatory fibers and dorsal root afferents that project to the L6 spinal cord. RESULTS: Volume/micturition in metabolic cages gradually increased in both contusion and transection groups compared with normals, and operated groups did not differ from each other. Urodynamic parameters from cystometry were significantly different in contusion and transection groups compared with normals, but again there was no significant difference between contusion and transection groups. Immunocytochemical analyses at the L6 spinal cord showed no serotonergic or noradrenergic fibers in transection group, but some descending fibers remained in contusion group, indicating sparing. Small dorsal root afferents were denser in both contusion and transection groups than in normals, indicating sprouting. CONCLUSIONS: Although differences were not found in LUT function in operated animals, supraspinal and dorsal root projections to the L6 spinal cord responded differently to contusion and transection. This suggests that the benefits of pharmacologic treatments may be different in two lesion models.

Preoperative renal scar as a risk factor of postoperative metabolic acidosis following ileocystoplasty in patients with neurogenic bladder.

OBJECTIVES: We investigated relation of preoperative renal scar to incidence of postoperative metabolic acidosis following ileocystoplasty in patients with neurogenic bladder. PATIENTS: Thirty patients with neurogenic bladder, who underwent ileocystoplasty, were enrolled in the present study. Median age at ileocystoplasty was 13.9 years and median follow-up period after ileocystoplasty was 8.2 years. Metabolic acidosis was defined based on the outlined criteria: base excess (BE) is less than 0 mmol l(-1). Preoperative examination revealed that no apparent renal insufficiency was identified in blood analysis, although preoperative (99m)Tc-DMSA scintigraphy indicated abnormalities such as renal scar in 14 patients (47%). Incidence of postoperative metabolic acidosis was compared between patients with and without preoperative renal scar, which may reflect some extent of renal tubular damage. RESULTS: Postoperative metabolic acidosis was identified in 13 patients (43%). Incidence of postoperative metabolic acidosis was significantly higher in patients with renal scar (11/14, 79%) compared with patients without renal scar (2/16, 13%; P<0.01). Particularly, all eight patients who had bilateral renal scars showed metabolic acidosis postoperatively. Compared with patients without preoperative renal scar, pH (P<0.05) and BE (P<0.01) were significantly lower postoperatively in patients with preoperative renal scar. However, there was no significant difference in PCO2. Hyperchloremia was observed in each patient with or without preoperative renal scar. CONCLUSION: Incidence of postoperative metabolic acidosis was significantly implicated in preoperative renal scar. If renal abnormalities are preoperatively identified in imaging tests, we need to care patients carefully regarding metabolic acidosis and subsequent comorbidities following ileocystoplasty.

Lysyl oxidase secreted by tumour endothelial cells promotes angiogenesis and metastasis.

BACKGROUND: Molecules that are highly expressed in tumour endothelial cells (TECs) may be candidates for specifically targeting TECs. Using DNA microarray analysis, we found that the lysyl oxidase (LOX) gene was upregulated in TECs compared with its expression in normal endothelial cells (NECs). LOX is an enzyme that enhances invasion and metastasis of tumour cells. However, there are no reports on the function of LOX in isolated TECs. METHODS: TECs and NECs were isolated to investigate LOX function in TECs. LOX inhibition of in vivo tumour growth was also assessed using ß-aminopropionitrile (BAPN). RESULTS: LOX expression was higher in TECs than in NECs. LOX knockdown inhibited cell migration and tube formation by TECs, which was associated with decreased phosphorylation of focal adhesion kinase (Tyr 397). Immunostaining showed high LOX expression in human tumour vessels in vivo. Tumour angiogenesis and micrometastasis were inhibited by BAPN in an in vivo tumour model. CONCLUSION: LOX may be a TEC marker and a possible therapeutic target for novel antiangiogenic therapy.

The incidence and mechanism of sunitinib-induced thyroid atrophy in patients with metastatic renal cell carcinoma.

BACKGROUND: To elucidate the incidence and mechanisms of sunitinib-induced thyroid atrophy, we investigated serial volumetric and functional changes, and evaluated histological changes of the thyroid gland in metastatic renal cell carcinoma patients who received sunitinib. METHODS: Thyroid volume (by computed tomography volumetry) and thyroid function were measured at baseline, during the treatment, and at post-treatment periods. Histological evaluation of the thyroid gland was performed in four autopsied patients. RESULTS: The median reduction rate in thyroid volume at last evaluation during sunitinib treatment was 30% in all 17 patients. The incidence of hypothyroidism during sunitinib treatment was significantly higher in the high reduction rate group (n=8; more than 50% reduction in volume) than in the low reduction rate group (n=9; less than 50% reduction in volume). Half of the patients in the high reduction rate group exhibited a transient thyroid-stimulating hormone suppression, suggesting thyrotoxicosis during sunitinib treatment. Histological evaluation demonstrated atrophy of thyroid follicles and degeneration of follicular epithelial cells without critical diminution of vascular volume in the thyroid gland. CONCLUSION: Thyroid atrophy is frequently observed following sunitinib treatment and may be brought about by sunitinib-induced thyrotoxicosis or the direct effects of sunitinib that lead to degeneration of thyroid follicular cells.

Prognostic significance of CD45RO+ memory T cells in renal cell carcinoma.

BACKGROUND: Memory T cells are well known to have a critical role for host defense in humans. However, their role in actual human cancer remains largely unknown. In this study, we tried to reveal the clinical importance of tumour-infiltrating CD45RO+ memory T cells in renal cell carcinoma (RCC). METHODS: We analysed 105 patients with RCC, who received radical or partial nephrectomy. Those were 65 in TNM stage I, 7 in stage II, 15 in stage III, and 18 in stage IV, respectively. CD45RO expression was evaluated by immunohistochemistry. CD4 and CD8 expressions were also systematically assessed in the same manner. RESULTS: Patients with higher TNM stage or high nuclear grade were found to have higher densities of CD45RO. Furthermore, CD45RO status was positively correlated with preoperative C-reactive protein level. In prognostic analysis, CD45RO+lo patients had a significantly better prognosis than CD45RO+hi patients. There was also a significant difference between CD4+lo and CD4+hi groups, whereas no significant difference was observed in CD8 T-cell status. Finally, multivariate analysis revealed that CD45RO+ status was the independent prognostic factor for patient overall survival. CONCLUSION: CD45RO+ memory T-cell status has a significant independent prognostic value, indicating that the adaptive immune response is functionally critical in human RCC.

Purified eicosapentaenoic acid induces prolonged survival of cardiac allografts and generates regulatory T cells.

Fish oil, which is rich in eicosapentaenoic acid (EPA), has been found to have immunomodulatory effects. We examined whether administration of purified EPA affected survival of fully mismatched murine cardiac allografts. Hearts from C57BL/10 (H-2(b)) mice were transplanted into CBA (H-2(k)) recipients treated with one intraperitoneal dose of purified EPA the day of transplantation. Untreated CBA recipients and recipients given 0.1 g/kg of EPA rejected C57BL/10 hearts (median survival time [MST], 8 and 13 days, respectively). With a 1.0 g/kg dose of EPA, graft survival was markedly prolonged (MST >100 days). To determine whether regulatory cells were generated, naïve mice (secondary recipients) underwent adoptive transfer of splenocytes from EPA-treated primary recipients and cardiac allograft transplantation. Adoptive transfer of whole, CD4(+) and CD4(+)CD25(+) splenocytes from EPA-treated recipients induced indefinite survival in secondary recipients. Flow cytometry showed that the CD4(+)CD25(+) cells were Foxp3(+). In reverse transcriptase-polymerase chain reaction (RT-PCR) studies, the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) mRNA was upregulated by EPA treatment. A PPARgamma antagonist abrogated the prolongation of graft survival induced by EPA treatment (MST, 13 days). Thus, in our model, purified EPA induced prolonged survival of fully mismatched cardiac allografts and generated regulatory T cells dependent on PPARgamma activation.

Low-density lipoprotein receptor activity in the guinea pig adrenal cortex. II. Zonal response to aminoglutethimide and 17 alpha-ethinylestradiol.

Low-density lipoprotein (LDL) receptor activity and the concentration of cholesterol were measured in the outer (glomerulosa/fasciculata) and inner (reticularis) zones of the adrenal cortex of the guinea pig to examine the relation between cholesterol content and LDL receptor activity. While the concentration of cholesterol was 2-3-times higher in the outer cortical zone, the maximum high-affinity binding capacity for LDL was essentially the same for the two zones, or slightly higher for the inner zone. Adrenocorticotrophic hormone (ACTH) caused a significant increase in LDL receptor activity only in the outer zone, but led to a reduction in the cholesterol content in both adrenocortical zones. The treatment of animals with 17 alpha-ethinyl-estradiol also resulted in a reduction of cholesterol in both adrenocortical zones, but an increase in LDL receptor number only in the outer zone. The latter effect was partially reversed by the administration of dexamethasone. Aminoglutethimide, which was used in a dose that did not block steroidogenesis but did block the hydrolysis of cholesteryl esters in response to ACTH, did not prevent the ACTH-induced increase in LDL receptor number in the outer zone. Thus, the number of LDL receptors was increased in the zona fasciculata by ACTH in the absence of a reduction in cellular cholesterol content, while the number of LDL receptors in the zona reticularis was not increased by ACTH even in the face of a reduction in cellular cholesterol. Exclusive of the experiments employing aminoglutethimide, when the cellular cholesterol content was plotted against LDL binding activity, an excellent inverse correlation was revealed for the zona fasciculata, but essentially no correlation was noted for the zona reticularis. It is concluded that the outer and inner cortical zones of the guinea pig adrenal are quite distinct in the nature of their LDL receptor activity and regulation: the LDL receptor of the outer zone appears to function in a way similar to what has been reported for the whole adrenal cortex of other species in that receptor number correlates with tissue cholesterol content and is primarily regulated by ACTH; the LDL receptor number of the inner zone, however, does not correlate with tissue cholesterol content and is apparently not regulated by ACTH.

Up-regulation of elafin/SKALP gene expression in psoriatic epidermis.

The expression of mRNA for elafin/SKALP, an inhibitor of leukocyte elastase and proteinase 3, in human normal and psoriatic epidermis was examined by in situ hybridization. In normal epidermis, elafin/SKALP mRNA was detected in the granular layer, but not in the spinous or basal layers. In fully developed psoriatic lesions, elafin/SKALP mRNA was found in the suprabasal layers of the ridges, and in the upper two thirds of the stratum malpighii at the elongated rete ridges. Intense staining was noted near the subcorneal microabscess in psoriasis vulgaris and under the subcorneal pustule in localized pustular psoriasis. In the marginal psoriatic epidermis, elafin/SKALP mRNA was expressed from the middle or upper spinous layer to the subcorneal layer, and the cells expressing elafin/SKALP mRNA increased especially under the parakeratotic corneal layer intermingled with pyknotic nuclei of neutrophils. These findings suggest that the induction of elafin/SKALP gene expression is related closely to the infiltration of neutrophils into the epidermis in psoriasis and plays an important role in protecting the skin components against the tissue damage caused by the infiltrated leukocytes.

Low density lipoprotein receptor activity in the guinea pig adrenal cortex. I. Zonal characterization and response to adrenocorticotropin.

The binding of [125I]iodo-low density lipoprotein ([125I]iodo-LDL) to isolated membranes was determined for the outer (glomerulosa/fasciculata) and inner (reticularis) zones of the guinea pig adrenal cortex. Binding of [125I]iodo-LDL to membranes from both zones was found to be highly specific and dependent on the presence of divalent cations, such as calcium. Saturation analysis revealed that the maximum high affinity binding capacity was similar for both zones under control conditions (approximately 400 ng/mg protein). When ACTH was administered, however, the maximum high affinity binding capacity increased in the outer zone by at least 3-fold, while there was no significant change for the inner zone. It was also noted that dexamethasone administration decreased [125I]iodo-LDL high affinity binding activity to less than half the control level in the outer zone without altering the binding activity in the inner zone. The cholesterol content in the outer zone was 2-3 times greater than that in the inner zone. When ACTH was administered, the concentration of cholesterol decreased significantly in both zones. Thus, ACTH appeared to influence cholesterol metabolism in the outer and inner adrenocortical zones, but was able to modulate LDL receptor activity only in the outer zone.

Postnatal development of [D-Ala2]deltorphin-I-like immunoreactive structures in the rat brain.

[D-Ala2]deltorphin-I, a highly selective ligand for delta opioid receptors, is a heptapeptide originally purified from frog skin. Previous immunohistochemical studies indicate that [D-Ala2]deltorphin-I-like molecule(s) may be present in adult rat brain, including specific neuronal cells and fibers partially overlapping with the mesocortical and nigrostriatal dopaminergic systems. Here, we examined the developmental aspect of such immunoreactive brain structures in early postnatal rats. In newborn to 21-day-old rats, positive staining in the brain occurred mainly in subpopulations of neurons and occasionally in tanycytes. On postnatal day 0, neuronal cell bodies containing [D-Ala2]deltorphin-I-like immunoreactivity were found in various brain regions, including the olfactory tubercle, ventral pallidum, hippocampus, ventral tegmental area, pars compacta of the substantia nigra, supramammillary nucleus, and dorsal raphe nucleus. Immunoreactive nerve fibers were observed in the main and accessory olfactory bulbs, olfactory tubercle, prelimbic area, anterior cingulate cortex, neostriatum, accumbens, lateral septal nucleus, lateral habenular nucleus, and superior colliculus. As pups grew, positive staining of cell bodies decreased gradually in both density and intensity, and those in the olfactory tubercle and ventral pallidum were no longer visible on postnatal day 14. On postnatal day 21, positive cells were found only in the ventral midbrain, including the pars compacta of the substantia nigra, ventral tegmental area, A8 region, and supramammillary nucleus. Positive fibers also decreased in density with age except in the accessory olfactory bulb, olfactory tubercle, prelimbic area, and anterior cingulate cortex.

Diminished penile expression of vascular endothelial growth factor and its receptors at the insulin-resistant stage of a type II diabetic rat model: a possible cause for erectile dysfunction in diabetes.

Erectile dysfunction (ED) is commonly experienced in men with diabetes mellitus. Vascular endothelial growth factor (VEGF) has been extensively documented for its pathogenic significance in different complications of diabetes. We hypothesized that expressions of VEGF, its receptors and its signaling pathway Akt may be drastically altered in diabetic penile tIssues and their alterations may modulate penile expression of the molecules that are believed to play a role in diabetic ED. Otsuka Long-Evans Fatty (OLETF) rats, a type II (non-insulin-dependent) diabetes mellitus, were used at the insulin-resistant stage of type II diabetes (20 weeks of age). We determined protein and mRNA expressions of VEGF, its receptors, Akt, nitric oxide synthase isoforms, and apoptosis-related molecules in the penis using immunohistochemistry, Western blotting, in situ hybridization, and real-time quantitative PCR analyses. The penile sections were also submitted to the Tdt-mediated dUTP nick end labeling assay for apoptosis. OLETF rats showed marked reductions in penile expression of VEGF, its two receptors and Akt. In OLETF rat penises, endothelial and neuronal nitric oxide synthase isoforms were expressed less abundantly. Furthermore, while anti-apoptotic markers, Bcl-2 and phosphorylated Bad, were down-regulated, pro-apoptotic markers, active caspase-3 and Bax, were up-regulated, resulting in the appearance of apoptotic cells in the penile tIssues of OLETF rats. The VEGF signaling system would work less well in diabetic penile tIssues as a result of the reduced expression, leading to diminished endothelial production of nitric oxide and apoptosis-related erectile tIssue damage. We propose that the abnormalities of the VEGF signaling system in the penis may play a role in the pathophysiology of diabetic ED.

Quantification of renal function with a count-based gamma camera method using technetium-99m-MAG3 in children.

UNLABELLED: To evaluate renal function quantitatively without blood sampling in children, renal uptake by gamma camera renography using 99mTc-MAG3 was compared with plasma clearance by a single blood sample method as the reference. METHODS: Twenty children (15 boys, 5 girls; aged 2-14 yr) with nephrourological diseases were examined prospectively in this study. The patient received an intravenous administration of 5 MBq/kg 99mTc-MAG3 which was prepared using a commercially available kit. Gamma camera renography was performed and the renal uptake per injected dose (%RU) of the 1-min period of postinjection was calculated from a background-corrected renogram curve by computer. The plasma clearance (Clmag) of 99mTc-MAG3 was calculated by the single blood sample method at 35 min postinjection. RESULTS: The %RU of the 1-min period in the 1-3 min postinjection correlated well with Clmag. The best correlation was observed 1-2 min postinjection. The regression equation between total %RU (X) and Clmag (Y) (ml/min/1.73 m2) was Y = -98.509 + 20.373X (r = 0.910, s = 84.19) with standardization by BSA. The best fit regression equation between individual %RU (X) and Clmag (Y) (ml/min/1.73m2) was Y = -43.799 + 19.917X (p = 0.932, s = 43.27). CONCLUSION: The renal uptake method based on separate counts by gamma camera renography using 99mTc-MAG3 does not require a blood sample for quantification of renal function and may be potentially more practical in children.

Distinctive glycolipid patterns in Wilms' tumor and renal cell carcinoma.

Glycolipid patterns were analysed chromatographically in Wilms' tumor and renal cell carcinoma tissues and compared with those of uninvolved tissue. Ganglioside GM3 was found to be increased in both cancer tissues, whereas sulfatides accumulated only in renal cell carcinoma, as reported earlier. Neolactotetraosylceramide was detected in both cancer tissues, but not in the uninvolved kidney tissues. In four cases of Wilms' tumors, only a low level of sulfotransferase towards galactosylceramide was found in one case, while no activity was detected in the three other cases. Present results show that the increased sulfatide(s) in the renal cell carcinoma and the deficiency of the sulfatides in Wilms' tumors appear to be biochemical characteristics of histologically different carcinomas.

Prophylactic chemotherapy with anthracyclines (adriamycin, epirubicin, and pirarubicin) for primary superficial bladder cancer. The Hokkaido University Bladder Cancer Collaborative Group.

A multicentric randomized trial was conducted to evaluate the efficacy of intravesical chemoprophylaxis for primary superficial bladder cancer. The 299 eligible patients with primary superficial bladder cancer were randomized into four groups (A, B, C, and D) after pathological confirmation. Intravesical instillation of drugs, which were dissolved in 20 ml physiological saline (PS; group A, 20 mg Adriamycin; group B, 20 mg epirubicin; group C, 20 mg pirarubicin; group D (control), PS alone], was performed once a week for 2 weeks after trasurethral resection and then once every 2 weeks for 14 weeks, once monthly for 8 months, and once every 3 months for 1 year. No significant difference in the patients' characteristics was found among the four groups. The follow-up period ranged from 3 to 31 months (mean, 14 months). The nonrecurrence rates were estimated by the method of Kaplan and Meier. The relative effects of five variables (the tumor status, size, grade, and stage and the treatment) on the efficacy of the chemoprophylaxis regimens were evaluated using a multiple regression model. Although the nonrecurrence rates determined for groups A and B were significantly higher than that found for group D (P < 0.05), no significant difference in the nonrecurrence rate was detected among groups A, B, and C. The multiple regression model indicated that the most important factors in preventing tumor recurrence at 12 or 24 months were the intravesical instillation of an anthracycline and the tumor status (solitary). These results demonstrate that intravesical instillation of the tested anthracyclines is effective for at least 2 years as prophylactic chemotherapy for primary superficial bladder cancer.

Normal perfusion pressure breakthrough syndrome in giant arteriovenous malformations.

The treatment of large, high-flow cerebral arteriovenous malformations is one of the most difficult operations which neurosurgeons encounter because of the complex surgery and the post-operative effects on the brain. We have evaluated 10 patients with large, high-flow AVMs who underwent surgical resection. Patients were investigated with contrast-enhanced computed tomography and magnetic resonance imaging, 1231-IMP single photon emission computed tomography (SPECT) studies of cerebral flow and cerebral vasodilatory function, intraoperative Laser Doppler flowmetry (4 or 10 patients), and conventional angiography. The volume of the arteriovenous malformation nidi ranged from 32.8 to 210.5 cc. SPECT imaging performed on the first post-operative day showed marked hyperperfusion in the brain tissue surrounding the resected nidus, and these regions were normal on images on the 7th post-operative day. Laser Doppler flowmetry showed sudden, and marked increase in CBF immediately following placement of temporary clips on the main feeding artery. Angiograms done on 7-14 days following surgery showed a stagnating artery, fragile vessel, and a prolonged circulation time. Our results indicate that pre- and post-operative SPECT study, especially a dynamic SPECT study done on the first post-operative day, was the most useful examination for ascertaining the post-operative normal perfusion pressure breakthrough.

Surgical treatment of internal carotid siphon aneurysms.

Surgical treatment of internal carotid artery aneurysms around the carotid siphon is discussed. The surgical approach to the aneurysms in this region, is as follows: 1. A fronto-temporal approach with the patient in a 45 degrees semi-sitting position to decrease venous pressure. 2. A Dolenc approach cutting a part of the dura mater of the superior orbital fissure to facilitate removal of the anterior clinoid process and unroofing of the optic canal. 3. Opening the medial triangle followed by transection of the optic canal dural sheath. Carotid siphon aneurysms can be divided into three groups anatomically; aneurysms of the ophthalmic segment (C2), those of the clinoid segment (C3), and those of the horizontal segment (C4). We present 29 cases of aneurysms arising from the C2 or C2/3 segment, 14 cases arising from the C3 or C3/4 segment, and 11 cases arising from the C4 segment. Anatomic localization of the aneurysms was established preoperatively by angiography and three-dimensional CT imaging. Small aneurysms of the ophthalmic segment projecting infero-medially can be clipped using a contralateral approach via the prechiasmatic root. Aneurysms of the ophthalmic segment projecting superiorly can be clipped following resection of the anterior clinoid process. The clinoid process should be resected intradurally with direct visualization of the aneurysms. Straight side-angled clips are suitable for these aneurysms. Carotid cave aneurysms, which include aneurysms of the ophthalmic segment oriented infero-medially and of the clinoid segment projecting postero-medially, can be clipped using curved fenestrated clips via Dolenc's extradural approach. For accurate clipping, opening of the medial triangle and full mobilization of the IC at the clinoid segment and optic nerve by unroofing the optic canal are required. Aneurysms of the horizontal portion are clipped after full exposure of the artery in the cavernous sinus only when the aneurysms are large and symptomatic. We used the fronto-temporal and Dolenc approaches and applied fenestrated clips to aneurysms oriented or postero-medially and straight or oblique clips to aneurysms projecting antero-laterally. Out of 40 aneurysms which underwent surgical clipping, 37 resulted in good post-operative recovery. There were three deaths secondary to complications of vasospasm and three cases with post-operative visual loss. The classification of these aneurysms and the surgical techniques we employed are discussed in detail.

Establishment of monoclonal antibody to human androgen receptor and its clinical application for prostatic cancers.

Hybrid cell lines were prepared by the fusion of BALB/c myeloma NS-1 cells with the lymphocytes of BALB/c mice that were immunized with partially purified androgen receptor (AR) from human prostates. One of the clones, 5F4, was chosen for detailed specificity analysis. The avidin-biotin-peroxidase complex (ABC) procedure was used for immunohistochemical staining of the 5F4 monoclonal antibody. In human benign prostatic hypertrophy (BPH) tissues, cytoplasm and nuclei were stained. Of 16 prostatic cancer tissues, 2 were composed of AR-positive cells exclusively, 7 were composed of AR-negative cells, and 7 contained both AR-positive cells and AR-negative cells (mixed). Of nine cases that were AR-positive or mixed, seven cases responded to the hormone therapy, and two were not determined for responsiveness because the patients died early of other diseases. Of seven AR-negative cases, all but one inestimable case had no response to the hormone therapy. Immunohistochemical analysis of AR by using the monoclonal antibody 5F4, was a useful tool for determining androgen dependency of prostatic cancers.

Comparative study of renal scintigraphy with 99Tcm-mercaptoacetyltriglycine and 123I-orthoiodohippurate.

This study was undertaken to compare image quality, renal handling and plasma clearance of 99Tcm-mercaptoacetyltriglycine (99Tcm-MAG3) with those of 123I-orthoiodohippurate (123I-OIH). 99Tcm-MAG3 was prepared in-house using an instant kit vial and 123I-OIH was commercially available in a form already labelled. Both studies were undertaken in 20 patients with nephrourological disorders after intravenous (i.v.) injection of 300 MBq 99Tcm-MAG3 and 34 MBq 125I-OIH within the time interval of 3 to 4 days. 99Tcm-MAG3 was much superior to 123I-OIH in visualization of renal perfusion, renal parenchyma and urinary tract. The time parameters of the renogram, Tmax and T1/2 were significantly prolonged in 99Tcm-MAG3 compared to those for 123I-OIH (P < 0.05). Plasma clearance of 99Tcm-MAG3 was also slower than that of 123I-OIH but correlated well with that of 123I-OIH (r = 0.958, P < 0.01). The % renal uptake of 99Tcm-MAG3 per injected dose, which was calculated by the gamma camera method, also correlated well with effective renal plasma flow (ERPF) calculated by the single blood sampling method (r = 0.940, P < 0.01). The renal handling and plasma clearance of 99Tcm-MAG3 are not completely identical with those of 123I-OIH. However, 99Tcm-MAG3 has the clinical advantages of superior image quality, the feasibility of gamma camera measurement of quantitative renal uptake, and ready availability for routine clinical use.