Simultaneous determination of purine and pyrimidine metabolites in HPRT-deficient cell lines.

Genetic mutations in the purine salvage enzyme, hypoxanthine-guanine phosphoribosyltransferase (HPRT), are known to cause Lesch-Nyhan syndrome and Kelley-Seegmiller syndrome. In patients, purine metabolism is different from that of normal persons. We have previously developed a method for simultaneously determining the concentration of purine and pyrimidine nucleosides and nucleotides. This system was applied to determine the concentrations of nucleosides and nucleotides in HPRT-deficient cell lines. The amount of inosine 5'-monophosphate (IMP) was different in Lesch-Nyhan syndrome, Kelley-Seegmiller syndrome, and control cell lines. The difference in the amount of IMP confirmed the mutation of the enzyme.

In-Vitro Simulation of NBCA Embolization for Arteriovenous Malformation.

SUMMARY: Embolization using n-butyl-cyanoacrylate (NBCA) for arteriovenous malformation (AVM) is now a daily practice over the world, but there exists no objective data that can be a basis for discussion or decision-making on the best concentration and injection rate of NBCA mixture. The purpose of this study was to obtain objective data on control and behavior of NBCA mixture with an in vitro simulation system of NBCA embolization for AVM. A nidus model made of a one-ml syringe filled with small beads was connected to a pulsatile flow circuit. A microcatheter was introduced just before the nidus model. Endoluminal pressures proximal and distal to the nidus and flow volume through the nidus were measured. Digital subtraction angiography (DSA) was performed to calculate transit time of the contrast medium (CM) through the nidus. NBCA was injected at various rates with an autoinjector and transit time of NBCA through the nidus was calculated. 27 trials were completed. Transit time of CM through the nidus model is well correlated to flow volume per unit of time through the nidus model. Shorter the transit time, larger was the flow volume per unit of time. The correlation was statistically significant (P < .0001). Though statistical significance was not attained, transit time of NBCA mixture at 50% concentration had a tendency to be correlated to flow volume per unit of time through the nidus, and slower injection of the NBCA mixture led to slower filling of the nidus model. Though this simulation system was artificial and the results should be interpreted carefully, it was shown with this system that transit time of CM through the nidus could be a good index for flow volume per unit of time through the nidus. Also suggested was a possibility to utilize this in vitro system for research and training on NBCA embolization of AVM.

Solitary schwannoma of the colon: report of two cases.

Some patients with gastrointestinal schwannoma (GIS) have been previously reported in the literature. However, GIS of the colon is quite rare. In addition, it is sometimes difficult to differentiate neurogenic tumors from other soft tissue tumors. We herein describe two cases of schwannoma of the colon, while also reviewing the relevant Japanese literature. The first case, a 73-year-old woman underwent a sigmoidectomy with lymph node dissection following the diagnosis of submucosal tumor. In the second case, a submucosal tumor was located in the cecum of a 44-year-old man. An endoscopic tumor resection was performed in the second case. The resected tumors measured 3.6 and 1.0 cm in maximal diameter, respectively. Microscopically, the tumors consisted predominantly of spindle-shaped cells that proliferated in an interlaced fashion. Mitosis was rarely seen in these tumors. Immunohistochemically, the tumor cells were strongly positive for S-100 protein, weakly positive for glial fibrillary acidic protein, and negative for CD34, alpha-smooth-muscle actin, and cytokeratin (CAM 5.2) in both cases. The tumors in the two cases were both diagnosed to be benign schwannoma of the colon. In general, schwannoma of the gastrointestinal tract is considered to be benign and should therefore be distinguished from other spindle-cell tumors or malignancies. Once diagnosed as schwannoma, extensive surgery should be avoided. Actually, such patients tend to show a good postoperative course with no evidence of recurrence.

Hepatoid adenocarcinoma of the stomach.

Although gastric cancer occurs frequently in Japan, few cases of hepatoid adenocarcinoma, a cancer with an extremely poor prognosis, have been reported. Here, we describe a 67-year-old Japanese man referred to our hospital with suspected gastric cancer. Gastrointestinal fiberscopy revealed an elevated lesion with a central depression on the lesser curvature, extending from the antrum to the body of the stomach. On the preoperative examinations, abdominal computed tomography scan, magnetic resonance imaging, and abdominal ultrasonography revealed multiple metastases to the liver and no cirrhotic change. The serum level of alpha-fetoprotein (AFP) was markedly elevated (10,084 ng/ml). After a diagnosis of AFP-producing gastric cancer with multiple liver metastases was made, total gastrectomy, without liver resection, was performed. Microscopically, the tumor showed two main histological features. The main part of the tumor resembled moderately differentiated hepatocellular carcinoma, and the rest showed fetal-type adenocarcinoma. Some parts of the hepatoma-like lesion showed periodic acid-Schiff (PAS)-positive granules. Furthermore, the tumor showed diffuse immunohistochemical positivity for AFP, alpha-1 antitrypsin, and alpha-1 antichymotrypsin. According to these histopathological findings, the tumor was diagnosed as hepatoid adenocarcinoma of the stomach. Although anastomotic leakage occurred postoperatively and the liver metastases have increased in size, the patient remains alive 11 months after the operation. Because of the poor prognosis for this histological type of tumor, accurate diagnosis of hepatoid adenocarcinoma is important, and long-term follow-up is required. We describe this rare case of hepatoid adenocarcinoma of the stomach, and review the literature concerning the clinicopathological aspects.

Dynamic gadolinium-enhanced MR imaging of pituitary adenomas: usefulness of sequential sagittal and coronal plane images.

Dynamic magnetic resonance (MR) imaging for pituitary adenomas is usually performed in a coronal direction; however, small lesions between slices, or lesions located at the anterior or posterior aspect of the pituitary gland might be overlooked on MR images in only the coronal direction. The purpose of our study was to evaluate whether consecutive dynamic MR images in the coronal and sagittal planes improve detection of pituitary adenomas. Eighteen patients with pituitary microadenomas and nine with healthy pituitary glands were included in this study. MR images were performed with 1.5 T superconductive units and commercially-available head coils. After a 5 ml gadolinium contrast injection, eight serial dynamic sagittal images were obtained. Within 3 or 6 min, this was followed by a 10-15 ml gadolinium injection and acquisition of eight serial dynamic coronal images. Dynamic MR images and conventional noncontrast- and contrast-enhanced sagittal and coronal T1-weighted images were evaluated independently in a blind fashion by two neuroradiologists regarding the depiction of pituitary microadenomas. The sensitivities of dynamic enhanced MR imaging in the detection of microadenomas were 61.1% in sagittal direction, 72.2% in coronal direction respectively, and were superior to those of conventional noncontrast- and contrast-enhanced T1-weighted imaging (22.2-50%). The sensitivity of a combination of sagittal and coronal dynamic enhanced MR imaging for the detection of microadenomas was 88.9% and was superior to those of conventional noncontrast- and contrast-enhanced T1-weighted imaging combining sagittal and coronal directions (61.1%, 61.1%) (P<0.05, P<0.05, respectively). The specificity and accuracy of dynamic enhanced MR imaging with combination of sagittal and coronal images was 88.9% respectively. Dynamic gadolinium-enhanced MR imaging, especially using both sagittal and coronal planes, was concluded to be useful for the detection of pituitary microadenomas.

Alternatively spliced MDM2 transcripts in human breast cancer in relation to tumor necrosis and lymph node involvement.

Several short forms of alternatively spliced Murine double minute 2 (MDM2) transcripts have recently been shown to correlate with high-grade malignancy in a number of human tumors. We examined the frequency of splice variants and their correlation with clinicopathological features in 60 cases of human breast cancer. Seven short forms coexpressed with wild-type mRNA were detected by nested RT-PCR. Sequencing of all the MDM2 variants demonstrated mRNA splicing which disrupted not only the conserved p53-binding domain but also, further towards the carboxy-terminus, the conserved nuclear localization sequence and/or the acidic and zinc finger domains. There was no significant correlation between the coexpression of splice variants and tumor size, histologic type or hormone (estrogen and progesterone) receptor status. However, cases with spliced MDM2 transcripts tended to be of a more aggressive type with axillary lymph node involvement and extensive necrosis in the tumors. Although the functional significance of MDM2 variants remains obscure, we anticipate that these variants will be confirmed as a novel prognostic marker in human breast cancer.

Amputation neuroma mimics common hepatic duct carcinoma.

Most amputation neuromas of the biliary tract occur in the cystic duct stump after cholecystectomy and are asymptomatic. However, when they arise in the main hepatic duct and are associated with obstructive jaundice, it is difficult to distinguish them from carcinoma. We describe a case in which preoperative differential diagnosis was difficult. A 60-year-old man was admitted to the Institute of Clinical Medicine, University of Tsukuba, with a chief complaint of jaundice. Cholangiography showed an irregularly elevated nodular lesion on the lateral wall of the common hepatic duct and multiple floating stones in the choledochus. Ultrasonography and computed tomography revealed one-sided regional thickening of the common hepatic duct associated with dilatation of the intrahepatic and extrahepatic bile ducts. Carbohydrate antigen 19-9 level was markedly elevated to 11,200 IU/mL in the bile juice, but was only 38 IU/mL in the serum, below the limit of abnormality. Cholangioscopy showed papillary tumor with coarse granular surface mimicking papillary carcinoma, but biopsy revealed no malignancy. The patient underwent hepaticocholedochus resection. Although the macroscopic finding from the surgical specimens was papillary carcinoma of the common hepatic duct penetrating to the hepatoduodenal ligament, histopathological examination revealed an amputation neuroma consisting of hypertrophic nerve tissues and giant cells containing foreign bodies, probably as a consequence of a previous cholecystectomy. The postoperative course was uneventful and the patient has been living well for the 5 years since the resection.

Assessment of hypermethylated DNA in two promoter regions of the estrogen receptor alpha gene in human endometrial diseases.

OBJECTIVE: DNA methylation in the promoter regions of many genes is associated with the regulation of gene expression. We examined the frequency of DNA hypermethylation at two nucleotide positions, the proximal promoter region (PPR) in exon 1 and the distal promoter region (DPR) in exon 1', of the estrogen receptor alpha (ERalpha) gene in 111 cases of various human endometrial diseases. METHODS: The degree of hypermethylation of PPR and DPR was examined by semi-quantitative competitive polymerase chain reaction assay using restriction enzymes (HpaII, NotI, and SacII). RESULTS: Endometrial tissues in the proliferative phase obtained from patients with leiomyomas and/or adenomyosis and no significant hormonal abnormalities did not show hypermethylation at the HpaII cleavage position of DPR. In 6 of 16 (37.5%) cases of simple endometrial hyperplasia, the PPR was hypermethylated, whereas in cases of atypical endometrial hyperplasia and endometrioid adenocarcinoma the frequencies were extremely low. Hypermethylation of these promoter regions did not correlate with lack of ERalpha protein in the 46 cases that were analyzed by enzyme immunoassay. CONCLUSIONS: ERalpha gene transcription from the distal promoter, rather than from the proximal promoter, is predominant in the proliferative phase of the normal menstrual cycle, because the PPR, but not the DPR, is frequently hypermethylated under those conditions. Conversely, ERalpha gene transcription from the proximal promoter is predominant in precancerous lesions such as atypical hyperplasia and endometrial cancer. However, hypermethylation at the promoter sites we examined was not related to the loss of ERalpha protein in endometrial disease.

Comparison of macroscopic appearance and estrogen receptor-alpha regulators after gene alteration in human endometrial cancer.

The existence of two types of endometrial cancer (hyperplasia-associated type [type I] and atrophy-associated type [type II]) is well established. To test if different molecular genetic pathways are involved in the pathogenesis of type I and II disease, we examined pathologic features and the genetic alterations of K-Ras, MDM2 and p53 (which are considered to be involved in regulation of the estrogen receptor-alpha) in human endometrial tissue samples using several modified PCR methods. We found a significant difference in histologic grade (P < 0.001), degree of invasion (P < 0.001), stage grouping (P < 0.001) and estrogen receptor status (P < 0.01) between type I and II cases. There was a tendency for cases with K-Ras point mutations to be of type I, and for cases with the p53 point mutation to be of type II; however, we found that these mutations or alternative splicing of MDM2 was rarely involved and there was no significant difference in frequency of these alterations between types I and II. There must therefore be another structural or functional difference of Ras, MDM2 or p53 between type I and type II cancer. These unknown factors may be responsible for the difference between estrogen-dependent and estrogen-independent growth in human endometrial cancer.

Overexpression of mitogen-activated protein kinase superfamily proteins unrelated to Ras and AF-1 of estrogen receptor alpha mutation in advanced stage human breast cancer.

Transactivation of the activation function-1 (AF-1) region of the estrogen receptor alpha (ER-alpha) gene is regulated by pathway "cross-talk" from Ras mitogen-activated protein kinase (MAPK). An analysis of this system is important for solving the problem of resistance to anti-estrogen agents used in the treatment of human breast cancer. We investigated the ER-alpha and Ras gene mutations and the MAPK-related protein status in 103 cases of breast carcinoma. None of the cases showed mutations in the AF-1 region of the ER-alpha gene. Despite the extremely low frequency of K- and H-Ras mutations in codon 12 (2/103 and 0/103), Ras p21 overexpression was identified in 29.1% (30/103), suggesting that the Ras activation in almost all cases we studied was not caused by point mutations but by enhanced expression. Our immunohistochemical analysis showed that the cases with overexpression of Ras and MAPK proteins (Ras p21, ERK-1, JNK-1, and p38) had a progressive tendency towards invasive growth, advanced-stage cancer, and decreased levels of ER-alpha protein. These results suggest that enhanced MAPK activity could be one of the characteristics of advanced breast cancer and that it could be involved in the transformation into estrogen-independent growth.

In vitro regioselective stability of beta-1-O- and 2-O-acyl glucuronides of naproxen and their covalent binding to human serum albumin.

beta-1-O- (NAG) and 2-O-glucuronides (2-isomer) of (S)-naproxen (NA) were prepared to determine which positional isomer(s) of the acyl glucuronide of NA is responsible for forming covalent adducts with human serum albumin (HSA). Their comparative stability and covalent binding adduct formation with HSA were investigated at pH 7.4 and at 37 degreesC. NA and its acyl glucuronides were simultaneously determined by HPLC. Three positional isomers were formed successively after incubation of NAG in the buffer only. However, when NAG was incubated with HSA (30 mg/mL), isomers other than the 2-isomer were formed in little or negligible quantities. In HSA solution, NAG (kd = 2.08 +/- 0.08 h-1) was four times less stable than 2-isomer (kd = 0.51 +/- 0.02 h-1). NAG was degraded by hydrolysis (khyd = 1.01 +/- 0.10 h-1) and isomerization (kiso = 1.07 +/- 0.07 h-1) to the same extent; however, hydrolysis was predominant for the 2-isomer (kd = 0.51 +/- 0.02 h-1). The incubation of both NAG and 2-isomer with HSA led to the formation of a covalent adduct; however, the adduct formation from the 2-isomer proceeded more slowly than that from NAG. The present results suggest that the covalent binding of NA to HSA via its acyl glucuronides proceeds through both transacylation (direct nucleophilic displacement) and glycation mechanisms; NAG rapidly forms an adduct that may be unstable, and the protein adduct from the 2-O-acyl glucuronide is as important for the covalent binding as those from the 1-O-acyl glucuronides.

MR angiography of cerebral vasospasm in preeclampsia.

Cerebral vasospasm was demonstrated with MR angiography in a patient with preeclampsia. MR angiography 5 days after the onset of symptoms clearly demonstrated diffuse intracranial vasospasm. The follow-up study confirmed the resolution of vasospasm successfully.

Triple primary brain tumors of different histological types: case report.

We report a case of triple primary intracranial tumors of different histologic types: meningioma, astrocytoma, and pituitary adenoma. Von Recklinghausen's disease or other etiologic factors supposedly associated with multiple primary brain tumors were not recognized in this 64-year-old female patient. Triple primary brain tumors with no association of von Recklinghausen's disease are rare--there is only one reported case with histologic studies.

Congenital subclavian steal syndrome with anomaly of the aortic arch.

A case of congenital subclavian steal syndrome associated with an anomalous right-sided aorta is reported. A 41-year-old man complaining of vertigo and a loss of consciousness was admitted. Physical examination revealed a blood pressure differential between the arms that was 20 mm Hg less in the left. Aortography showed a right aortic arch from which arose the right subclavian and both common carotid arteries. The left subclavian artery did not opacify. Right vertebral angiography showed retrograde filling of the left vertebral. A left carotid-subclavian bypass was performed. Postoperatively, the patient is symptom free with equalized blood pressure.

[A midbrain arteriovenous malformation at quadrigeminal plate completely obliterated by embolization].

An eighteen year-old boy presented sudden loss of consciousness and tetraparesis. Radiological examinations revealed a ruptured midbrain arteriovenous malformation (AVM) at the right quadrigeminal plate. The patient recovered from symptoms by conservative treatment over 6 months with residual right hemisensory disturbance and mild diplopia during bilateral horizontal gaze. One year later, he was admitted to our institute for radical treatment of the AVM. On the angiography, the AVM was fed by two branches of the right long circumferential artery and a left paramedian penetrating artery of the posterior cerebral artery (PCA), and drained into the straight sinus via a dilated quadrigeminal vein. After much discussion on several strategies, endovascular surgery was chosen as a radical treatment. In the initial session, a medial branch of the right long circumferential artery was catheterized with Tracker-18, and embolized with 1.0 ml of ethylene vinyl alcohol copolymer (EVAL) without any deficits after negative provocative tests using Amytal and Xylocaine. Then, a lateral branch of the same artery was embolized with 0.8 ml of EVAL in the same way without any deficits. In the second session, catheterization into the left paramedian artery was difficult because of its small diameter and steep branching angle, and only a 3 mm hooked tip of Tracker-18 could be canulated into the feeding pedicle. Left oculomotor palsy was induced by provocative Xylocaine test. However, angiographic cure of the AVM was expected to be made possible by the occlusion of this pedicle, and the procedure was continued due to the request of the patient and his family.(ABSTRACT TRUNCATED AT 250 WORDS)

Endovascular coil embolization of a recurrent giant internal carotid artery aneurysm via the posterior communicating artery after cervical carotid ligation: case report.

The case of a 29-year-old man with a giant fusiform aneurysm of the left internal carotid artery (ICA) is presented. The aneurysm, treated by cervical ICA ligation and extracranial-intracranial bypass, recurred 4 years later owing to recruitment of the posterior communicating artery (PCoA). Because of the previous bypass surgery a direct surgical approach was excluded. After an initial failure with balloon embolization, the aneurysm was embolized successfully with occlusive platinum microcoils through the microcatheter navigated into the aneurysm via the enlarged PCoA. Endovascular coil embolization may be useful in the treatment of cerebral aneurysms not amenable to direct surgery or balloon embolization.

[Association of an epidermoid tumor with a cerebral aneurysm on the contralateral internal carotid artery].

A 71-year-old female was evaluated for a sudden loss of consciousness. Initial computed tomography and cerebral angiography revealed a severe subarachnoid hemorrhage, a large cystic lesion in the left occipital region, and an aneurysm at the bifurcation of the internal carotid and the posterior communicating artery on the right side. As the subarachnoid hemorrhage was thought to be due to rupture of the aneurysm, we attempted to clip the aneurysmal neck at first, and 2 weeks later the cystic tumor was removed for the purpose of internal decompression. The extirpated specimen proved to be an epidermoid tumor. Association of a brain tumor and an cerebral aneurysm is very rare, especially a combination of an epidermoid tumor and a cerebral aneurysm, and until now only 2 cases have been reported to the best of our knowledge. Moreover, the epidermoid tumor of our case was located intracerebrally. The phenomenon that the internal carotid aneurysm was on the left side and the tumor on the right side may suggest that these 2 lesions originated quite coincidentally. It is controversial to decide which lesion and when to treat first in a case of association of a tumor and an aneurysm, and we discussed as to the surgical therapy reviewing literature.

[Angiographic findings of vertebral dissecting aneurysm. Report of two cases and review of literature].

The authors report two cases of vertebral dissecting aneurysm. The first case, a 49-year-old female, developed severe headache and computed tomography scan showed subarachnoid hemorrhage (SAH), but 4-vessel cerebral angiography failed to show an aneurysm. The second angiograms obtained 2 weeks later showed possible aneurysmal dilatation on the right vertebral artery. The third angiograms, 2.5 months after SAH, disclosed a right vertebral fusiform aneurysm on the arterial phase and it was diagnosed as a dissecting aneurysm since the contrast medium remained until the very late venous phase. The previous angiograms were reviewed using the subtraction technique, which revealed retention of the contrast medium. The second case, a 42-year-old female, suffered from SAH. Left vertebral angiography revealed a fusiform aneurysmal tapered narrowing just distal to the aneurysm, which was a typical "pearl and string sign." The subtraction film of the venous phase also showed retention of the contrast medium in the aneurysmal portion. These findings accurately diagnosed dissecting aneurysm of the vertebral artery. Since the classical true diagnostic "double lumen sign" was rarely observed in the angiograms, it was not easy to diagnose dissecting aneurysm of the vertebral artery. The authors emphasize the angiographic findings of retention of the contrast medium in the venous phase as a "true diagnostic sign" for correct diagnosis of dissecting aneurysm.

[Inhibition of virus multiplication by immunoactive peptides]. / Inhibition de la multiplication des virus par des peptides immunoactifs.

In the present study we show that peritoneal macrophages obtained from the mice treated with the immunoactive peptides inhibit the multiplication of Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), but not that of vesicular stomatitis virus (VSV), and that the intraperitoneal administration of the peptides suppresses the infection with HSV-1 in mice.

Application of "Hirt supernatant" DNA to the molecular epidemiology of cytomegalovirus infections.

Human cytomegalovirus DNA was isolated from infected cells by the Hirt method (B. Hirt, J. Mol. Biol. 26:365-369, 1967). The restriction endonuclease cleavage patterns of DNA obtained in this manner were comparable with those of DNA extracted from purified virions. The "Hirt supernatants" were satisfactory for identifying individual cytomegalovirus strains by their DNA fingerprinting.