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PubMed1997

[Genetic diagnosis for post-transfusion graft-versus-host disease using PCR amplification of variable number of tandem repeats].

Post-transfusion graft-versus-host disease (PT-GVHD) is a potentially fatal adverse reaction to blood transfusion with a mortality rate of 95 percent or higher. We previously developed a definitive diagnostic test for PT-GVHD using polymerase chain reaction (PCR) amplification of highly polymorphic microsatellite repeat sequences. The results show the replacement of patient original DNA types with donor types on development of PT-GVHD. However, samples of patient blood obtained before transfusion are rarely stored until more than 2 weeks after the transfusion when PT-GVHD is usually suspected. On these occasions, we analyzed PCR amplified fragments of DNA extracted from fingernail clippings to determine the patient original DNA types. As a rapid and easily method, this system could be a useful tool for the clinical diagnosis of PT-GVHD.

Graft vs Host Disease↗
PubMedDate not supplied

Interaction between longevity-associated mitochondrial DNA 5178 C/A polymorphism and cigarette smoking on hematological parameters in Japanese men.

Mitochondrial DNA 5178 C/A (mt5178 C/A), namely NADH dehydrogenase subunit 2 237 Leu/Met, polymorphism is as reported in literature associated with longevity and susceptibility to ischemic heart disease or cerebrovascular disorders in the Japanese population. Previous reports suggested that mt5178A genotype exerts antiatherogenic effects. The aim of this study was to investigate whether mt5178 C/A polymorphism is associated with hematological parameters, such as thrombogenic risk factors for myocardial infarction and stroke, in 321 healthy Japanese men. No significant differences were observed between mt5178 C/A genotypes, but in subjects with body mass index (BMI) of < or = 23, this polymorphism influenced the effects of habitual smoking on hematological parameters. Red blood cell (RBC) counts were significantly lower and mean corpuscular hemoglobin (MCH) levels were significantly higher in smokers with mt5178A than nonsmokers with mt5178A. Platelet counts were significantly higher in smokers with mt5178C than nonsmokers with mt5178C. Cigarette consumption was strongly associated with RBC counts, mean corpuscular volume levels, and MCH levels for men with mt5178A, and was associated with platelet counts for those with mt5178C. Moreover, BMI was significantly positively associated with RBC counts and platelet counts only in men with mt5178A, age was significantly negatively associated with RBC counts only in men with mt5178C. These data suggest that mt5178 C/A polymorphism may influence the effects of cigarette smoking on hematological parameters in healthy BMI < or = 23 Japanese men.

Age Factors↗
PubMed1981

[Basic examination on the energization of experimental animal feces by methane fermentation (author's transl)].

Basic experiments of methane fermentation of livestock excreta have been reported by many investigators, yet there has been no report of experiments of methane fermentation of experimental animal feces. We considered that it would be feasible to gasiby experimental animal feces by methane fermentation. In this report, methane gas fermentation of rat feces is studied. The results are summarized as follows: 1. Methane fermentation of rat feces yielded, under the conditions described, 220-4231 of gas per kg of organic matter. 2. Decomposition ratio of BOD removal volume in organic matter was 50-60% when the fermentation condition was optimum. 3. The volume of gas from BOD removal volume in organic matter was 500-600ml/g. 4 Methane gas contents in the gas generated from the feces ranged from about 55-60%. 5. COD removal ratio was above 80%, and BOD removal ratio above 90%. From these data, the authors conclude that under the optimum conditions, described sufficient methane gas can be obtained from rat feces by methane fermentation.

Animals↗
PubMed1988

[Limb salvage surgery for pelvic malignancies followed by reconstruction with hip endoprosthesis].

Between 1983 and 1987, nine patients with pelvic malignancy were treated with limb salvage surgery and preoperative chemotherapy. They were 6 males and 3 females, ranging in age from 13 to 73. There were 4 chondrosarcomas, 3 osteosarcomas and 2 Ewing's sarcomas. Seven patients received preoperative chemotherapy mainly consisting of adriamycin and cis-platinum. The remaining 2 patients with Ewing's sarcoma received T-11 protocol by Rosen. Surgical margins were wide but partially marginal in 6 patients and intra-lesional in 3 patients. Reconstruction with hip endoprosthesis fixed with screws and bone cement in the sacrum was performed after whole involved iliac bone was resected. Local recurrence was detected in one patient with osteosarcoma treated by marginal resection and 2 patients treated by intralesional resection. Complications occurred in 5 patients. Dislocation was observed in the first patient in this series. Other complications were infection in one patient, sciatic nerve palsy in one patient and leg length shortening in 2 patients. One patient walks without crutches, 5 patients with them. One patient uses a wheelchair and 2 patients are on bed rest. Four patients survived with no evidence of disease for 8-51 months (average 25 months). Two patients have survived with disease for 11-16 months (average 14 months). Three patients died from metastasis a 6-10 months (average 9 months).

Adolescent↗
PubMed1993

Assessment of membrane fusion efficiency and its use for distinguishing epitopes on the fusion (F) protein of Sendai virus (HVJ).

Upon incubation of radioiodinated Sendai virus with Ehrlich ascites tumor cells to induce viral envelope-cell membrane fusion, TCA-soluble radioactivity was found to be rapidly released from the virus into the medium. The amount of the TCA-soluble 125I species released into the medium was correlated with the extent of envelope-membrane fusion. Using a new method based on this phenomenon, we evaluated the fusion-inhibiting activity of monoclonal anti-F protein antibodies and could distinguish epitopes within partially overlapping regions of the F protein by their effects on fusion activity. We concluded this method is a reliable and sensitive one for measuring envelope-membrane fusion.

Animals↗
PubMed2002

Method for analysis of 4-hydroxy-2-(E)-nonenal with solid-phase microextraction.

A simple analytical method for 4-hydroxy-2-(E)-nonenal (HNE) using solid-phase microextraction (SPME) fiber was developed. HNE or the derivative of HNE formed by reaction with 2,4-dinitrophenylhydrazine (DNPH) was extracted from the sample solution by immersing the SPME fiber into the solution, and the amount of HNE was quantified by HPLC. The extraction conditions of HNE and HNE-DNPH were examined, using standard solutions, with respect to fiber coating, NaCl concentration, rate of stirring, adsorption temperature, and adsorption time. The recovery of HNE reached 80%, and the quantification limits of HNE and HNE-DNPH using standard compounds were 14.1 pmol/10 mL and 486.5 fmol/10 mL, respectively. This method can be applied to the detection of HNE in oxidized oil or samples of porcine liver.

Aldehydes↗
PubMed1980

Varicella dendritic keratitis.

A 7-year-old boy and a 3-year-old girl suffered from unilateral disciform keratitis and iritis associated with varicella. While they were treated with topical corticosteroid, idoxuridine, and atropine drops, dendritic lesions typical of herpes zoster appeared four months after the onset of eruptive skin lesions. Using the direct immunofluorescent method, we showed varicella-zoster virus antigen in the epithelial cells scraped from the dendritic lesion.

Betamethasone↗
PubMed2001

Intravascular US-guided direct intrahepatic portacaval shunt with a PTFE-covered stent-graft: feasibility study in swine and initial clinical results.

PURPOSE: To determine the feasibility of the creation of a direct intrahepatic inferior vena cava (IVC)-to-portal-vein shunt with puncture guided by a transfemorally placed intravascular ultrasound (IVUS) probe and use of a polytetrafluoroethylene (PTFE)-covered stent-graft. MATERIALS AND METHODS: In five swine, transjugular access was used to perform a direct puncture from the IVC to the portal vein with use of a modified Rosch-Uchida Portal Access set directed with real-time IVUS (9 MHz) introduced from a transfemoral venous approach. The direct intrahepatic portocaval shunt (DIPS) was then created with single or overlapping PTFE-covered Palmaz stents placed through a 10-F sheath and dilated to a diameter of 8 mm. Follow-up was performed with transhepatic portography at 2, 4, and 8 weeks. Animals were killed when shunts occluded or at the termination of the study at 8 weeks. Gross and microscopic histologic study was performed on sacrificed animals. A similar technique was used to create DIPS in five patients with intractable ascites, with follow-up by US and venography. RESULTS: All experimental DIPS created in swine were created without complications. Portal vein punctures were achieved in four of five swine on the first or second pass of the needle. Follow-up transhepatic portography at 2 weeks demonstrated occlusion of two shunts, both explained by technical reasons at sacrifice. At 4 and 8 weeks, the remaining three shunts were patent on portography. Histology showed a thin neointimal lining with no significant tissue ingrowth or hyperplasia. Clinically, in five patients, successful puncture of the portal vein from the IVC was achieved in one to three passes. Creation of DIPS led to a reduction of mean portosystemic gradient from 18-29 mm Hg (mean, 24 mm Hg) to 9-10 mm Hg (mean, 9 mm Hg). One patient died of liver failure 2 days after creation of DIPS. The other four patients were doing well 2-15 months (mean, 8 months) after the procedure, with patency confirmed by US and venography. CONCLUSION: Creation of DIPS is technically feasible, and the direct IVC-to-portal-vein puncture can be done accurately with real-time IVUS guidance. Further studies and longer follow-up are necessary to determine if the short length of the PTFE-covered stent-graft and avoidance of the hepatic vein will increase the long-term patency compared to standard transjugular intrahepatic portosystemic shunt creation.

Animals↗
PubMed2000

Serological diagnosis of idiopathic pulmonary alveolar proteinosis.

Previously, we reported the specific occurrence of neutralizing autoantibodies against granulocyte-macrophage colony-stimulating factor (GM-CSF) in the bronchoalveolar lavage fluid from 11 Japanese patients with idiopathic pulmonary alveolar proteinosis (I-PAP). The autoantibody was also detected in sera from all 5 I-PAP patients examined. To determine that the existence of the autoantibody is not limited to the Japanese patients, we examined sera from 24 I-PAP patients in five countries and showed that the autoantibody was consistently and specifically present in such patients. Thus, detection of the autoantibody in sera can be used for diagnosis of I-PAP. To establish a simple and convenient method for diagnosis of I-PAP, we developed a novel latex agglutination test using latex beads coupled with recombinant human GM-CSF. GM-CSF binding proteins isolated from the sera using the latex beads were identified as the autoantibodies of IgG(1) and IgG(2). The titer of the autoantibody determined by this test correlated with that determined by ELISA. Agglutination was positive in 300-fold diluted sera from all 24 I-PAP patients, but negative in sera from four secondary PAP patients, two congenital PAP patients, 40 patients with other lung diseases, and 38 of 40 normal subjects. These results establish that the latex agglutination test is a reliable method for serological diagnosis of I-PAP with high sensitivity (100%) and specificity (98%).

Adult↗
PubMed1997

Enhanced myopathy following administration of hypolipidemic agents under urethane anesthesia.

The enhanced effect of urethane anesthesia on the serum creatine kinase (CPK) level following administration of hypolipidemic agents was examined to develop a convenient experimental screening method for drug-induced myopathy. After oral administration of a hypolipidemic agent to rats, 25% urethane solution was infused intravenously at a very low rate using a microinfusion pump. Blood samples were collected 7 h after drug administration and the risk of myopathy was evaluated based on the CPK level. When bezafibrate (BF), simvastatin (SV), or pravastatin (PV) (50-500 mg/kg) was orally administered under urethane infusion, enhanced elevation of the serum CPK level was observed dose dependently for BF and SV, but not for PV. The elevation of serum CPK was much higher with BF than with SV or PV. In addition, when SV or PV (50-500 mg/kg) was coadministered with 50 mg/kg of BF, there was a striking increase in the serum CPK level as compared with the drug alone. Without urethane infusion, no significant elevation in serum CPK level was observed even at a high dose of these hypolipidemic agents. These phenomena suggest that the urethane anesthesia enhanced the elevation of the serum CPK level following administration of hypolipidemic agents. We propose that this method is a simple and speedy screening test for drug-induced myopathy.

Anesthesia↗
PubMed2000

Genome-wide profiling of gene amplification and deletion in cancer.

Accumulations of genetic changes in somatic cells induce phenotypic transformations leading to cancer. Among these genetic changes, gene amplification and deletion are most frequently observed in several kinds of cancers. Amplification of oncogene and/or deletion of tumor suppressor gene, together with dysfunction of the gene by point mutation, are the main causes of cancer. Genome-wide analysis of amplification and deletion of genes in cancers is basic to resolving the mechanisms of carcinogenesis. Comparative genomic hybridization (CGH) developed in 1992 has been utilized to identify DNA copy number abnormalities in various kind of cancers and several reports have shown its usefulness in screening of the genes involved in carcinogenesis, and also in the identification of prognostic factors in cancer. We have shown that 1q23 gain is associated with neuroblastomas that are resistant to aggressive treatment, and have poor prognosis, and 1q and 13q gains are possibly related to drug resistance in ovarian cancers. Recently, the "rough draft" of the human genome was reported and we are ready to utilize the vast information on genomic sequences in cancer research. Moreover, microarray technology enables us to analyze more than ten thousand genes at a time and revealed genetic abnormalities in cancers at a genome-wide level. By combination of microarray and CGH, a powerful screening method for oncogenes and tumor suppressor genes in cancers, called array-CGH, has been developed by several groups. In this article, we overview these genome-wide analytical methods, CGH and array-CGH, and discuss their potential in molecular characterization of cancers.

Gene Amplification↗
PubMed2003

[Non-radioactive PCR southern method for analysis of CTG repeat in myotonic dystrophy].

Myotonic dystrophy (MyD) is a hereditary neuromuscular disorder of an autosomal dominant trait. MyD is caused by an expansion of unstable CTG trinucleotide repeat in the 3' untranslated region of mRNA coding myotonin protein kinase (MT-PK). We analyzed CTG repeat expansion in 10 patients with congenital MyD and their relatives using the non-radioactive PCR Southern method. The region containing the CTG repeat was amplified by PCR using specific primers. The PCR products were electrophoresed on a 1% agarose gel and transferred to a nylon membrane. The CTG repeat expansion was shown using a fluorescein-labelled (CTG) 10 probe. To estimate the number of CTG repeats, we compared the smears obtained on Southern blotting with a picture of PCR products and a DNA size marker (100 bp). We compared our results of radioactive Southern blotting for genomic DNA digested by Eco RI or Bgl I and for PCR products. In congenital MyD patients, heterogeneous smears (3.89-10.22 kb:about 1252-3362 CTG repeat) were observed, whereas in the adult type MyD had heterogeneous smears (0.92-1.82 kb:about 262-562 CTG repeat). In asymptomatic MyD patients, there were heterogeneous smears (0.35-1.16 kb:about 72-342 CTG repeat). These results demonstrated anticipation. We conclude that the non-radioactive PCR Southern method is useful and convenient for the DNA diagnosis of MyD.

Adult↗
PubMed1995

[Y-grafting using the inferior epigastric artery for coronary revascularization].

On view of the remarkable good results obtained with arterial grafts for coronary revascularization, the authors continue to conduct the total arterial bypass grafting. However, the following disadvantages are encountered; 1) arterial graft crossing over the aorta and 2) left internal thoracic artery (LITA) is on some occasions impossible to anastomose with the left anterior descending (LAD) coronary artery. To eliminate these problems, "IEA Y-grafting" which the IEA is anastomosed with the LITA in an end-to-side fashion was conducted. From January to December 1994, 24 patients underwent coronary revascularization by this method. All 24 IEA grafts were anastomosed to the circumflex coronary artery (22), right coronary artery (3) or diagonal branch. (1). LITA graft were used on 24 patients (29 anastomosis) with the right gastro-epiploic artery (RGEA) at 16 (17) and RITA at 2. Immediate postoperative angiographic evaluation of the IEA grafts indicated 17 anastomoses to be patent (17/19, 89.5%). Two patients died due to coronary spasm. Although hypoperfusion poses a serious problem in this method, IEA may be considered to serve as a 3rd alternative for an arterial conduit.

Aged↗
PubMed1993

[Percutaneous hot water injection therapy (PHoT) for hepatic tumors: a clinical study].

Percutaneous ethanol injection therapy (PEIT) has become widely used in the treatment of HCC. However, the indications for this method are limited by the toxicity of ethanol itself. Against this background, the authors turned their attention to the heat coagulation necrosis effect induced by boiled physiological saline (hot water) and devised percutaneous hot water injection therapy (PHoT) as a new local treatment method. PHoT was performed a total of 41 times in 13 patients (16 nodules) with HCCs measuring < or = 3 cm. Changes in AFP values, CT findings, angiographic findings before and after treatment, and histopathological findings of needle biopsy or resected specimen were investigated. AFP values decreased in all of the seven patients who initially showed high values. On CT, all lesions receiving PHoT became hypodense, with this change thought to indicate necrosis. Disappearance of the tumor stain was confirmed in the four patients in whom follow-up angiography was performed. In the six patients in whom needle biopsy was performed, disappearance and scaring of tumor cells were observed, while in the one patient in whom the tumor was resected, complete necrosis of the tumor was confirmed. PHoT, despite a small number of punctures, shows good anti-tumor effects, and has promise not only as a curative local treatment method for small HCCs but also in combination with TAE for large HCCs.

Aged↗
PubMed1993

[A case of surgery for AAE with dissection extended to right coronary artery--device for reconstruction of coronary artery and retrograde cardioplegia and retrograde cerebral perfusion].

We experienced a surgical case of aortic dissection (Stanford type A) with annuloaortic ectasia in Marfan Syndrome. A 45-year-old female who had been diagnosed as Marfan Syndrome three years age was emergently admitted to our hospital with sudden chest pain. We diagnosed this case as aortic dissection of Stanford type A by ultrasonic cardiogram and thoracic CT. The ascending aorta was replaced with composite graft by modified Bentall procedure, right coronary artery was bypass to segment 2 using vein graft and left coronary artery was interposed by Dacron graft. During operation we utilized retrograde cold blood cardioplegia for cardiac arrest (180 min.) and retrograde cerebral perfusion accompanied with total circulatory arrest (45 min.). No complication such as cardiac damage and cerebral damage was found postoperatively. We discussed surgical method for aortic dissection with anuloaortic ectasia in particular reconstruction of coronary artery, cardioplegia and cerebral protection.

Aortic Dissection↗