Rapid and simple detection of PCR product DNA: a comparison between Southern hybridization and fluorescence polarization analysis.

The essential aim of this study was to compare two different methods, Southern hybridization and fluorescence polarization (FP) assay. They both detect specific hybridization and were examined using common asymmetric PCR products and probes. FP assay clearly showed the hybridization of probe DNAs with the asymmetric PCR products of their target genes. Southern blot patterns presented excellent consistency with the results of FP assay. In both methods, two types of Shiga toxin (vero toxin) genes held in enterohaemorrhagic Escherichia coli (EHEC) were used as target genes. For detection of the two genes, stx1 and stx2, two respective DNA probes were synthesized. Both in FP assay and in Southern hybridization, the probe for stx1 hybridized only with the product of stx1 and vice versa. The results of the DNA detection using different methods were completely in agreement. Moreover, FP assay makes it possible to detect the hybridization rapidly. In our high NaCl concentration condition, hybridization between the probes and the asymmetric PCR products could be monitored within about 15min.

Measurement of the time course of DNA/RNA hybridization and RNA detection using fluorescence polarization analysis.

Using fluorescence polarization analysis, the time courses of hybridization between probe oligo-DNAs and target RNAs were measured. The RNAs were amplified using the DNA templates of Shiga toxin genes by NASBA (Nucleic Acid Sequence Based Amplification). Two DNA probes were designed for detecting the genes and they rapidly and specifically hybridized with their target RNA sequences. NASBA could be sufficiently used for the combination and DNA/RNA hybridization could be detected in the fluorescence polarization.

Cancer mortality of thorotrast patients in Japan: the second series updated 1998.

The 150 male patients exposed to Thorotrast who were confirmed by a 1975-1978 national survey with diagnostic X rays of 50,860 war-wounded soldiers were followed up between 1979 and 1998 (Aichi series or second series). Age-adjusted rate ratios of deaths from all causes were 3.0 times higher in Thorotrast patients compared to controls; this was statistically significant. Rate ratios for liver cancer, liver cirrhosis and leukemia were 35.0, 7.5 and 18.2, respectively.

Summary of entire Japanese thorotrast follow-up study: updated 1998.

Updated data from two series in a cancer mortality study for a total of 412 Japanese Thorotrast patients were combined. The rate ratio for all deaths of Thorotrast patients, compared to controls, started to increase after a latent period of 20 years after injection of Thorotrast. Rate ratios for liver cancer, liver cirrhosis, leukemia and lung cancer were 35.9, 6.9, 12.5 and 2.0 times higher, respectively, than those for controls.

Type II citrullinemia associated with neutropenia.

A 37-year-old Japanese man was admitted with delirium and hyperammonemia. He was diagnosed as having type II citrullinemia because of an elevated citrulline level on amino acid analysis and very low hepatic argininosuccinate synthetase activity. He also showed a low neutrophil count and a low serum level of granulocyte colony-stimulating factor. Reduced production of this cytokine and/or impairment of its feedback regulation by the neutrophil count may have played a role in the neutropenia of this patient.

Endoscopic MRI using 3D-spoiled GRASS (SPGR) sequence for local staging of rectal carcinoma.

OBJECTIVE: Magnetic resonance imaging was performed with an MR endoscope to evaluate the depth of invasion of rectal carcinomas. MATERIALS AND METHODS: Twenty-two patients with rectal carcinomas were studied prior to surgery with MRI using a 3D-fast spoiled GRASS (SPGR) sequence, a nonmagnetic endoscope and an antenna probe. RESULTS: The MR images on 3D-fast SPGR sequence showed that the normal bowel mucosa had a high signal intensity, the submucosal layer had a low signal intensity, and the muscularis propria had moderate signal intensity. Small tumors had moderate signal intensity similar to the muscularis propria. Advanced tumors with considerable volume revealed heterogeneous findings with intermediate and low signal intensities. The depth of wall invasion by rectal carcinoma was correctly staged with endoscopic MRI in 16 of 22 patients. CONCLUSION: This technique may be useful for the accurate staging of the depth of invasion by rectal carcinomas.

[Early diagnosis of breast cancer].

Despite the increasing incidence of breast cancer in recent years, early detection of breast cancer improves the curative rate and quality of life (QOL). Advances in diagnostic imaging are making early detection increasingly common. Although mammography (MMG) is the most popular examination method and yields excellent image description, diagnosis is influenced by imaging technique and ability to read the images. A clear image can be achieved if the MMG image acquired at 28 Kv using an M0 filter is energy subtracted at 32 Kv using an Rh filter. Ultrasonography can be excellent for qualitative diagnosis of a tumor, but changes in blood flow volume, as demonstrated by Doppler imaging, makes early detection difficult. Breast scintigraphy by 201TlCl can clearly demonstrate palpable breast cancer and also lymph node metastasis, but poor spatial resolution remains a problem. High resolution CT is effective for detection of lymph node metastasis. MRI offers excellent qualitative diagnosis of tumors, but detection of ultra-small lesions is difficult even using contrast medium. The detection rate and proportion of early breast cancer detected on the whole will rise if MMG is introduced into the present mass screening for breast cancer, but its implementation requires improved facilities, imaging capability, and experience in image reading.

Hepatic arterial infusion chemotherapy for liver metastases from breast cancer.

Between 1985 and 1992, 56 patients with unresectable liver metastases from breast cancer were treated by repeated hepatic arterial infusion chemotherapy employing an implantable port system. 5-Fluorouracil (5-FU) at 330 mg/m2 per week. Adriamycin (ADR) at 20 mg/m2 every 4 weeks, and mitomycin C (MMC) at 2.7 mg/m2 every 2 weeks were given to 42 patients. The remaining 14 patients received 5-FU at 330 mg/m2 per week and epirubicin (EPIR) at 20 mg/m2 every 2 weeks. As a rule, the treatment was performed on an outpatient basis. The side effects and complications observed included myelosuppression (41%), hepatic arterial occlusion (23%), and gastric mucositis (20%), but no major toxicity was encountered. The response rate (CR+PR) of the evaluated patients as determined from CT scans was 81%. The overall median survival period was 12.5 months. Only 14% of the patients died due to regrowth of liver metastases, and in 70% of the total cases, death due to liver metastases was avoided by this treatment. Thus, repeated hepatic arterial infusion chemotherapy for liver metastases from breast cancer might be capable of prolonging the survival of patients via avoidance of death due to the liver metastases.

[Changes in arterial blood flow patterns by patients' posture during hepatic arterial infusion chemotherapy assessed by 99mTc-MAA perfusion scintigraphy].

Hepatic arterial perfusion scintigraphy (HAPS) with 99mTc-macroaggregated albumin (MAA) was performed to reveal the effect of patients' posture for the arterial blood flow patterns in hepatic arterial infusion (HAI) chemotherapy. 20 patients of liver metastases with the percutaneously implanted catheter into the hepatic artery underwent HAPS following administration of 99mTc-MAA at the supine and the upright position under one week interval. 10 ml of the saline with radionuclide was injected at a rate of 1 ml per minute. Acquired two images at the different position of each patient were compared and grouped into three classes; similar, slightly different and different. Of 20 patients, 6 (30%) were judged as similar, 8 (40%) as slightly different, and 6 (30%) as different. These results suggest that the drug distribution can change by patients' posture in case of the slow rate HAI performed with continuous infusion pump.

[Transurethral microwave thermotherapy for benign prostatic hyperplasia].

Transurethral microwave thermotherapy (TUMT) of prostate was administered to 10 patients with bladder outlet obstruction due to benign prostatic hyperplasia. The mean age of the patients was 74.4 years (range 63 to 85). The Prostatron device, which provides microwave heating of the prostate and conductive cooling of the urethra was used, and the prostate was heated with a calculated intraprostatic temperature of 45.5 degrees C for 55 minutes. No anesthesia was required for most of the patients. The clinical effects were evaluated at 4-6 weeks and 3 months after treatment. The symptomatic scores improved in the majority of patients. There was no significant change in prostate volume. The maximum flow rate and average flow rate were increased at 6 weeks and 3 months, but there was no significant change. The only side effects were transient hematuria and short-term obstruction secondary to urethral edema. In comparing TUMT with the transurethral resection of prostate (TUR-P), the maximum flow rate after TUMT was lower than that after TUR-P and the improvement of residual urine after TUMT was lower than that after TUR-P.

[Pharmacokinetics in arterial infusion chemotherapy].

The pharmacokinetics of arterial infusion chemotherapy was introduced. The pharmacokinetic rationales were based on the theories of "first pass effect" and "increased local concentration without first pass effect" advocated by Dr. Collins. These theories can offer much information to select the drug and to determine the administration schedule. However, for most of the drugs, the clinical pharmacokinetic studies are poor, and the optimal dose in bolus intraarterial infusion and the administration schedule is uncertain in terms of the dosage and hours. There are also some differences between pharmacokinetic understanding and clinical effect. Thus, further pharmacokinetic investigations and clinical studies are required to establish effective regimens for arterial infusion chemotherapy.

[Intermittent hepatic arterial infusion of high-dose 5-FU for liver metastases from colorectal cancer].

A phase II study of weekly intermittent hepatic arterial infusion of high dose 5-FU for liver metastases from colorectal cancer was carried out. Thirty-two patients with unresectable liver metastases from colorectal cancer were entered in this study. Nausea (< or = grade 2) was observed in 31% of the cases, mild elevation of rGTP in 13%, biloma in 9%, and hepatic arterial occlusion in 22%. However, no major toxicity occurred. The response rate evaluated by CT-scans was 75% (4 CR + 20 PR/32), and the overall median survival was 22 months. Significant differences of median survival were observed in the extra-hepatic lesions [extrahepatic lesions: (-) 16 months vs (+) 22 months]. This regimen showed high activity without major toxicity and the reduction of pts' QOLs caused by pumps. It confirmed by a study involving a large number of cases, this regimen will become a standard regimen for unresectable liver metastases from colorectal cancer.

pH mapping in living tissues: an application of in vivo 31P NMR chemical shift imaging.

Chemical shifts were extracted from in vivo 3-dimensional 31P NMR CSI data and pH images were constructed. The images could spatially resolve tissue pH ranging from 5.8 to 7.2 (with uncertainty of 0.11-0.17 pH unit) in an ischemia-reperfusion model of diabetic rat calf muscles.

[CT appearance of thorotrast-related and non-thorotrast-related peripheral cholangiocarcinoma].

To assess the differences in CT findings among patients with peripheral cholangiocarcinoma with and without a history of Thorotrast administration, CT studies from 13 Thorotrast patients and eight non-Thorotrast patients were reviewed. Diagnostic and prognostic differences were evaluated between the two groups. Despite periodic imaging surveillance, eight of the 13 (62%) lesions discovered by CT were larger than 6 cm. The prognosis for Thorotrast patients was unfavourable due to difficulties in early detection and complications from associated hepatic fibrosis. The main problem with early detection was that a background of uneven Thorotrast deposits visualized in the liver disguised the tumor as Thorotrast granulations. Although non-Thorotrast patients were not monitored regularly, they had a better chance of undergoing curative resection for the following three reasons: 1) it was easy to detect the tumor (detection rate, 100%); 2) this group rarely had associated liver cirrhosis in noncancerous areas, and 3) non-Thorotrast patients were younger than Thorotrast patients. Early detection of cancer by CA19-9 assay and imaging in asymptomatic subjects without any history of liver disease could be important steps toward the early and radical resection of cancer to achieve a better prognosis.

Detection of locally recurrent colorectal cancer with radiolabeled monoclonal antibody H-15.

H-15 (HT-29-15) is an IgG1 mouse monoclonal antibody (mAb) to a cell surface antigen (molecular mass, 200,000 daltons) present on virtually all colorectal cancers and also in normal pancreatic ducts and bile ducts, but not in other normal tissues. The biological distribution and imaging characteristics of iodine-131 (131I)-labeled mAb H-15 were studied in 5 primary colorectal cancer patients and 9 patients with local recurrence of colorectal cancer. H-15 mAb labeled with 0.5-10 mCi of 131I was administered 7 to 8 days before surgery at 4 dose levels, ranging from 0.2 to 6 mg. Selective mAb H-15 localization to tumor tissues was demonstrated in 6 of 12 patients with antigen-positive tumors: in two patients, recurrent tumors were negative to H-15 mAb, although the primary tumors were positive. In six patients with positive radioimaging, tumor:normal tissue ratios ranged from 2.05 to 5.35 and tumor:serum ratios from 1.18 to 2.73. The clarity of images seems to correlate well with the latter ratios. Technetium-99 (99mTc)-albumin blood pool studies in selected cases showed that local recurrence of colorectal cancers was hypovascular, emphasizing the selective localization of mAb H-15 despite poor blood flow distribution in the tumors. The results altogether demonstrated that radioimmunodetection with 131I mAb H-15 is valuable for differentiating recurrent colorectal cancer from granuloma formation after surgery.

Chemical assessment of phospholipid and phosphoenergetic metabolites in regenerating rat liver measured by in vivo and in vitro 31P-NMR.

For the assessment of 31P-NMR spectroscopic data, phospholipid precursors (phosphorylethanolamine (PE) and phosphocholine) and catabolites (glycerophosphorylethanolamine (GPE) and glycerophosphorylcholine (GPC)), as well as adenosine phosphates were chemically determined in regenerating rat liver. The data were compared with those obtained by in vivo and in vitro 31P-NMR spectroscopies. Chemical assay revealed a significant increase of PE and a decrease of GPE, GPC and ATP in hepatectomy group compared to sham operation group. The values obtained by in vitro NMR were in good agreements with those of chemical assay, but significant differences between the two groups were observed only in PE and inorganic phosphate (Pi). Noticeable increase in PME was not detected by in vivo 31P-NMR spectroscopy, although the increase of PE was about 2.5-times that of the control and its constitution ratio to the whole phosphomonoester (PME) was less than 15%. On the other hand, in vivo NMR showed a large phosphodiester (PDE) peak occupying approx. 40% of the total phosphorus signal, while the contribution of its constituents, GPE and GPC was about 5% found by both chemical assay and in vitro NMR. The PDE peak in in vivo NMR seemed to reflect the membrane phospholipid itself rather than its catabolites. A slight decrease of phosphoenergetic level in regenerating rat-liver was commonly suggested by all three analytical methods.

Clinical evaluation of assistant diagnostic system for mammograms using the auto-analyzing method.

Clinical usefulness of a newly developed auto-analyzing system of mammograms to search the abnormal dense areas such as cancer lesions was studied. 345 breasts including 86 cancers, 52 mastopathies, 18 fibroadenomas, 13 cysts, 4 lipomas and 172 normals were analyzed and compared with final diagnosis. 84 of 86 cancer lesions were correctly pointed out (sensitivity: 97.9%). Based on the gradient of the density, the ranking of the cancer or calcified lesions among suspected regions in a mammogram was decided. 55 lesions of them (64.0%) were ordered at the first rank and 83 (96.5%) were in the upper three ranks. 68 of 74 breasts with calcifications were correctly diagnosed and 48 of them were ranked in the upper three ranks. Although further improvement of the system is needed, this system will be a useful assistance for screening the breast cancer in the analyses of mammograms.

Management of patients with unresectable liver metastases from colorectal and gastric cancer employing an implantable port system.

Between 1985 and 1990, 50 patients with unresectable liver metastases from colorectal cancer and 34 subjects with metastases from gastric cancer were treated by repeated hepatic arterial infusion chemotherapy employing an implantable prot system. A catheter was inserted into the hepatic artery via the left subclavian artery and was connected to the implantable injection port in each patient. 5-Fluorouracil (5-FU) at 330 mg/m2 per week (167 mg/m2 daily given continuously over the initial 3 months for colorectal cancer), Adriamycin (ADR) at 20 mg/m2 every 4 weeks and mitomycin C (MMC) at 2.7 mg/m2 every 2 weeks were given to all 34 patients with gastric cancer and to 31 of the colorectal cancer patients. The remaining 19 patients with colorectal cancer received 5-FU at 1,000 mg/m2 every week. As a rule the treatment was performed on an outpatient basis. The side effects and complications observed included myelosuppression (23%), hepatic arterial occlusion (21%), and gastroduodenal mucositis (12%), although no major toxicity was encountered. The response rate (CR+PR) among the evaluated patients as determined using CT scans was 67% for colorectal cancer and 73% for gastric cancer. The overall median survival was 12 months and 15 months, respectively. Good local control of liver metastases from the colorectal and gastric cancers was achieved by repeated hepatic arterial infusion chemotherapy employing an implantable port system without the need for hospitalization and without producing major toxicity. Thus, the implantable port system is very useful for the management of patients with unresectable liver metastases.

[Outpatient cancer chemotherapy employing implantable systems].

Outpatient cancer chemotherapy (OCC) employing implantable systems was introduced and the objects, conditions and problems of OCC were discussed based on experiences in 324 cases. The aims of OCC are improved QOL and the continuation of chemotherapy. Our requirements are safety, effectiveness, easy management and non-disturbance of activity. Implantable systems are very useful for OCC, especially continuous infusion combined with ambulatory pumps. However, the improvement of ambulatory pumps and the establishment of methods to evaluate OCC are required to further develop OCC.

[Mass screening of breast cancer with imaging diagnosis].

Westernization of our life-style and diet has caused a higher incidence of breast cancer in Japan. Although mass screenings such as anamnesis, visual and palpable examinations are provided by local government in Japan, no curable cancer in the early stage can be detected without the use of imaging diagnosis. Furthermore, the image can be saved as a record. Mammography yields images that are best suited for breast cancer examination in terms of sensitivity, specificity and accuracy. However, the large volume of information collected from the images makes diagnosis extremely difficult. In order to ease the burden on the diagnosing physician and to reduce the examination cost, CRT diagnosis by digital images and image selection by an automatic diagnosis supporting system with neuro-computer must replace the present filming method. On the other hand, 100% accuracy can not be achieved by either mammography or ultrasonic examination due to the limitation in depicting images on a display monitor. The accuracy rates of mammography and ultrasonic examination are 91.5% and 87.5% in our hospital. Imaging diagnosis, however, must be efficiently applied in addition to the visual and palpable examinations in promoting mass screening for breast cancer.