Biomedical subjects

T Kohno

Publications and source records attributed to T Kohno.

At least 127 records · Page 7Linked to original sources

PubMed1999

Correlation between the status of the p53 gene and survival in patients with stage I non-small cell lung carcinoma.

The association of p53 abnormalities with the prognosis of patients with non-small cell lung carcinoma (NSCLC) has been extensively investigated to date, however, this association is still controversial. Therefore, we investigated the prognostic significance of p53 mutations through exons 2 to 11 and p53 protein expression in 103 cases of stage I NSCLC. p53 mutations were detected in 49 of 103 (48%) tumors. Two separate mutations were detected in four tumors giving a total of 53 unique mutations in 49 tumors. Ten (19%) of mutations occurred outside exons 5-8. Positive immunohistochemical staining of p53 protein was detected in 41 of 103 (40%) tumors. The concordance rate between mutations and protein overexpression was only 69%. p53 mutations, but not expression, were significantly associated with a shortened survival of patients (P<0.001). Furthermore, we investigated the correlation between the types of p53 mutations and prognosis. p53 missense mutations rather than null mutations were associated with poor prognosis (P < 0.001 in missense mutations and P=0.243 in null mutations). These results indicated that p53 mutations, in particular missense mutations, rather than p53 expression could be a useful molecular marker for the prognosis of patients with surgically resected stage I NSCLC.

Alleles↗

PubMed1999

Microsatellite instability in lymphoid leukemia and lymphoma cell lines but not in myeloid leukemia cell lines.

Microsatellite instability (MSI) represents a defect in the DNA mismatch repair system and has been shown to take part in the genesis and/or progression of several human malignancies. In hematological malignancies, the relevance of MSI has been a matter of controversy. Therefore, 29 microsatellite loci were examined for MSI in 57 leukemia and lymphoma cell lines by PCR analysis. Ladder formation of bands representing MSI was observed at multiple loci in 6 of 24 lymphoid leukemia/lymphoma cell lines and in 0 of 33 myeloid leukemia cell lines. Analysis for the BAT-26 and BAT-25 loci confirmed the presence of MSI in five of six lymphoid cell lines exhibiting ladder formation of bands. Thus, at least 5 out of 24 (21%) lymphoid leukemia/lymphoma cell lines were considered as being MSI-positive. These results indicate that MSI contributes to the development of lymphoid but not to myeloid malignancies.

Humans↗

PubMed1999

Pharmacokinetics of 9alpha-fluoromedroxyprogesterone acetate in rats: comparison with medroxyprogesterone acetate.

Medroxyprogesterone acetate (MPA) is widely used in endocrine therapy for breast cancer and other diseases. Recently, it has been demonstrated that 9alpha-fluoromedroxyprogesterone acetate (FMPA) also has anti-tumour activity in chemical-induced rat mammary tumour and its activity is greater than that of MPA. In the present study, the physico-chemical properties of FMPA and MPA and their pharmacokinetics in female rats were investigated. Partition coefficients (log P) of FMPA and MPA were 3.1 and 3.8, respectively, while the solubilities of FMPA and MPA in phosphate buffer saline were 3.8 and 1.1 microg/mL, respectively. When the two agents were intravenously or orally administered into female rats, there was no significant difference between their plasma concentrations. However, unmetabolized drug excreted into urine accounted for 4.7 and 0.7% of the intravenous dose of FMPA and MPA, respectively. The free fraction of FMPA in rat plasma was approximately four times that of MPA. Assuming the well-stirred model, hepatic intrinsic clearances of FMPA and MPA were estimated to be 64 and 293 L/h per kg, respectively. In addition, the free fraction of FMPA in blood is estimated to be higher than that of MPA, which may explain the higher anti-tumour activity.

Administration, Oral↗

PubMed1999

13CO(2) peak value of L-[1-(13)C]phenylalanine breath test reflects hepatopathy.

BACKGROUND: Using a rat model of hepatectomy, we investigated whether the severity of hepatopathy could be quantitatively measured from changes in expiratory (13)CO(2) levels after intravenous administration of L-[1-(13)C]methionine or L-[1-(13)C]phenylalanine. MATERIALS AND METHODS: Under nembutal anesthesia, 30 mg/kg L-[1-(13)C]phenylalanine or 40 mg/kg L-[1-(13)C]methionine was administered to rats through the femoral vein, and expiratory (13)CO(2) levels were measured for 15 min. A 30, 70, or 90% hepatectomy was performed. In the control group, simple laparotomy was performed. Breath test was conducted 20 min after laparotomy. We examined the correlation of the total (13)CO(2) output over 15 min or peak (13)CO(2) level with liver weight/body weight (%). RESULTS: In breath test graphs, L-[1-(13)C]methionine did not show any peak level during measurement. L-[1-(13)C]phenylalanine showed a specific peak level 6 +/- 1 min after administration. The correlation coefficient between total (13)CO(2) output over 15 min after L-[1-(13)C]methionine administration and liver weight/body weight was 0.922 (P < 0.001). The correlation coefficient between total (13)CO(2) output over 15 min after L-[1-(13)C]phenylalanine administration and liver weight/body weight was 0.883 (P < 0.001). The correlation coefficient between peak L-[1-(13)C]phenylalanine level and liver weight/body weight was highest, 0.927 (P < 0.001). CONCLUSION: In a breath test with intravenously administered L-[1-(13)C]methionine or L-[1-(13)C]phenylalanine, hepatopathy could be quantitatively evaluated by measuring expiratory (13)CO(2) levels over 15 min. After administration of L-[1-(13)C]phenylalanine, hepatopathy could be quantitatively evaluated in a short period by measuring the peak expiratory (13)CO(2) level.

Animals↗

PubMed1999

One-step affinity purification of the G protein betagamma subunits from bovine brain using a histidine-tagged G protein alpha subunit.

An efficient one-step affinity purification of bovine brain G protein betagamma subunits (betagamma's) is described. The betagamma's, in a detergent extract of brain membranes, are first dissociated from the alpha subunits (alpha's), reassociated with decahistidine-tagged alphail produced in bacteria, and then adsorbed onto Ni2+-nitrilotriacetic acid-agarose via the histidine tag. This mild adsorption retained the high activity of the ligand alpha's, in contrast to the commonly used chemical crosslinking methods. A wash step with a buffer containing chaotropic ions (SCN-) completely removed contaminating proteins that were refractory to washes with high concentrations of detergents, after which the highly purified betagamma's were eluted with a buffer containing Al3+, Mg2+, and F- ions. The obtained betagamma's were found to be fully functional, as assessed by their ability to support pertussis toxin-catalyzed ADP-ribosylation of alphail. Since the combination of the mild adsorption via the histidine tag and the wash with chaotropic ions can be easily applied to purifying betagamma's from various animal tissues, this new chromatographic method is expected to facilitate the purification of other membrane proteins that bind to Galpha and/or Galphabetagamma.

Adsorption↗

PubMed1999

Expression of CD10 on lymphoid cells associated with thymoma.

BACKGROUND: The thymoma is a mixture, in varying proportions, of epithelial and lymphoid cells. The aim of this study was to detect immature T-lineage components of lymphoid cells infiltrating thymoma tissue. METHODS: Tissue-infiltrating lymphoid cells (N = 10) and normal thymocytes (TC, N = 3) were retrieved from surgical specimens. The surface antigens of these lymphocytes were examined with flow cytometry. RESULTS: CD3 was detected on 48 +/- 19% of tissue-infiltrating lymphoid cells and 66 +/- 16% of TC, while CD19 was expressed on neither. CD4 and CD8 were co-expressed on 70 +/- 21% of Tissue-infiltrating lymphoid cells and 68 +/- 28% of TC. CD10 was expressed on 22 +/- 10% of tissue-infiltrating lymphoid cells, but was not expressed on TC (5 +/- 3%, p = 0.0003). Two-color analysis showed that the CD10+ fraction was weakly stained with CD3. It was also stained with anti-CD38 and anti-CD4, but not with anti-CD34. CONCLUSIONS: Tissue-infiltrating lymphoid cells included CD10-positive T-lineage cells. This fraction corresponds to very immature thymocyte subsets. These observations suggest that the epithelial component of a thymoma either inhibits the normal differentiation of T-lineage cells, or attracts immature T-lineage cells of bone-marrow origin.

Adult↗

PubMed1999

Fetal polycystic kidney disease in oro-facio-digital syndrome type I.

We report a girl with oro-facio-digital syndrome type I (OFD I) associated with polycystic kidney disease (PKD), which was identified on fetal US and fetal MRI. After birth, the diagnosis of this X-linked dominant disorder, which is lethal in males, was achieved by recognition of facial dysmorphism, lingual hamartomas, postaxial polydactyly, brain malformations, and the existence of her deceased male sibling with similar malformations. Adult PKD is a common feature in heterozygous females with OFD I. However, fetal PKD has been reported only in a lethal homozygous male. Our observation expands our knowledge about the phenotypic variations of PKD in OFD I.

Female↗

PubMed1999

Transthoracoscopic needle biopsy for indeterminate lung nodules.

BACKGROUND: The usefulness of transthoracoscopic needle biopsy for preoperatively indeterminate intrapulmonary nodules was evaluated. METHODS: Thoracoscopy was performed on 38 patients with pulmonary solitary nodules suspected to be primary lung carcinomas. When the nodule was localized by simple observation or tactile sensor, a biopsy specimen of the tumor was obtained by a biopsy needle introduced through a trocar. RESULTS: The nodules were 7 to 55 mm in diameter. All were located in the peripheral region of the lung. Biopsy specimens were obtained even from 17 nodules with no associated pleural changes. By cytology, all the malignant tumors were precisely diagnosed, 29 as primary lung cancers and 3 as metastatic lung neoplasms. Five of the remaining six benign nodules were not precisely diagnosed. However, they were cytologically classified as class I. CONCLUSIONS: Transthoracoscopic needle biopsy is feasible for diagnosing small intrapulmonary nodules, particularly those of malignant neoplasms. As compared with thoracoscopic excisional biopsy, transthoracoscopic needle biopsy saves time and may reduce the possibility of tumor dissemination during the procedure.

Adult↗

PubMed1999

Hemodynamic effects of carbon dioxide insufflation under single-lung ventilation during thoracoscopy.

BACKGROUND: The hemodynamic effects of carbon dioxide insufflation under single-lung ventilation were studied in 22 consecutive thoracoscopic harvests of the left internal mammary artery, which was used for minimally invasive coronary artery bypass grafting. METHODS: An electrocardiograph, arterial catheter, Swan-Ganz catheter, and transesophageal echocardiograph were used to monitor seven hemodynamic variables. Baseline data were obtained during ventilation of both lungs and the measurements were repeated after the left lung was collapsed and at 5 and 30 minutes after hemithorax insufflation with low-flow (2 to 3 L/minute) carbon dioxide gas was begun. The intrapleural pressure was maintained at 8 to 10 mm Hg. RESULTS: Thoracoscopic harvest of the internal mammary artery was completed in all cases with a mean insufflation time of 44+/-12 minutes. There were no significant changes in the mean arterial pressure, heart rate, cardiac index, and left ventricular ejection fraction throughout the procedure, whereas the central venous pressure, mean pulmonary arterial pressure, and pulmonary capillary wedge pressure (p < 0.05 for each variable) during insufflation. CONCLUSIONS: Low-flow carbon dioxide insufflation into the left hemithorax with an intrapleural pressure of 8 to 10 mm Hg under selective right-lung ventilation does not compromise the human heart with normal to moderately depressed function and can be an efficacious adjunct in specific thoracoscopic procedures.

Aged↗

PubMed1999

Age-related scattered hypofluorescent spots on late-phase indocyanine green angiograms.

PURPOSE: Scattered hypofluorescent spots may be seen on indocyanine green (ICG) angiograms of regions that do not show abnormalities when viewed with an ophthalmoscope. Hypofluorescent spots are found in several pathologic conditions, typically in inflammatory diseases. In this report, we describe hypofluorescent spots in normal fundi and show that such spots can be age-related. METHODS: Video-fundus camera ICG angiograms of 115 eyes of 109 patients aged 12 to 85 years with normal fundi or with only age-related maculopathy were reviewed. The relation between age and scattered hypofluorescent spots, and between age-related maculopathy and spots was examined utilising regression analysis. RESULTS: Scattered hypofluorescent spots were seen throughout the posterior pole in 24 eyes of 23 patients and in a portion of the posterior pole in 30 eyes of 29 patients. The hypofluorescent spots were noted between 26 and 37 minutes after dye injection. Patient age ranged from 51 to 80 years, and regression analysis showed that the frequency of hypofluorescent spots increased significantly with aging (p < 0.05). However, age-related maculopathy did not show a significant relation to the spots. CONCLUSION: Scattered hypofluorescent spots seen in the posterior pole during the late-phase of ICG angiograms can apparently be due to aging of the fundus.

Adolescent↗

PubMed1999

Mutation and expression of the p51 gene in human lung cancer.

A newly identified gene, p51, is a functional and structural homologue of the p53 gene and thus a Candidate tumor suppressor gene. To elucidate the role of the p51 gene in lung carcinogenesis, we determined the sequences of exon-intron boundaries and the 5'- and 3'-flanking regions of all the 15 coding exons and performed a mutation analysis, as well as detailed analysis for gene expression. A frameshift mutation was detected in 1 of 44 lung cancer cell lines, whereas no mutation was detected in 45 primary lung cancers. Thus, p51 mutation occurs only in a small subset of lung cancer. Expression of the p51 gene was detected in 23 of 43 cell lines by Northern blot analysis and 34 of 44 by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Thus, p51 expression is low or absent in a subset of lung cancer. The deltaN isotype of p51 transcripts was dominantly expressed in several cell lines, particularly in cell lines with high levels of p51 expression. Because the deltaN isotype encodes a protein that transdominantly suppresses the transactivation function of the TA type of p51, it is possible that p51 protein is not functionally active, even in lung cancer cells with p51 mRNA expression, due to expression of dominant-negative p51 protein. These results suggested that the p51 gene is inactive in a considerable proportion of lung cancers. RT-PCR analysis also revealed the presence of a novel type of mRNA transcript, p51delta, which lacks exons 12 and 13 by alternative splicing. The delta isotype was expressed in 18 of 44 lung cancer cell lines and in diverse normal tissues. Further analysis on p51 expression in cancerous as well as noncancerous cells will provide us with valuable information for the understanding of multiple functions of the p53 family proteins in human carcinogenesis.

Base Sequence↗

PubMed1999

Familial gastric cancer: clinicopathological characteristics, RER phenotype and germline p53 and E-cadherin mutations.

Gastric cancer frequently occurs in family members with hereditary non-polyposis colorectal cancer (HNPCC) and Li-Fraumeni syndrome (LFS) and germline E-cadherin mutations were recently identified in a subset of familial gastric cancers. Thus, families with an aggregation of gastric cancers were recruited by reviewing the genealogical trees of 3632 patients with gastric cancer. The criteria for recruiting such families were the following: at least three relatives should have gastric cancer and one of them should be a first degree relative of the other two; at least two successive generations should be affected; in one of the relatives gastric cancer should be diagnosed before age 50. Thirty-one cases (0.9%) fitted all three of these criteria. There were only gastric cancer patients in 18 of the 31 families and there were no families that fitted clinical criteria of HNPCC or LFS. Paraffin-embedded tissues were available in 29 probands and DNA was successfully isolated for molecular analyses in 13 probands. RER phenotype was detected in three (23%) cases, whereas germline p53 mutations were detected in none of 13 cases. A germline E-cadherin mutation was detected in one of three diffuse types and none of 10 intestinal types, however, a mutation resulting in the replacement of Gly by Val was detected in the precursor sequence. Thus, although familial clustering of gastric cancer occurs in approximately 1% of gastric cancer patients, germline mutations of the DNA mismatch repair, p53 and E-cadherin genes do not significantly contribute to such a clustering.

Adolescent↗

PubMed1999

How many tumor suppressor genes are involved in human lung carcinogenesis?

To date, only a limited number of tumor suppressor genes have been identified as being inactivated in lung cancer. The p53 and RB genes are frequently inactivated by genetic alterations such as chromosomal deletions and loss-of-function mutations, while the p16 gene is inactivated not only by genetic alterations but also by transcriptional silencing due to hypermethylation. Recently, it was shown that the FHIT gene encompassing the chromosomal fragile site, FRA3B, is also inactivated in a large proportion of lung cancers. Several lines of evidence indicate the presence of additional tumor suppressor genes involved in lung carcinogenesis. Lung cancer cells often show deletions at multiple chromosomal regions, and deletion mapping studies have defined more than 30 regions dispersed on 21 different chromosome arms as candidate tumor suppressor loci. Several chromosomal regions hypermethylated in lung cancer cells and a number of chromosomal fragile sites have been mapped to the regions deleted in lung cancer. These chromosomal loci can harbor unknown tumor suppressor genes inactivated in lung cancer. Studies on the inherited susceptibility to lung cancer in mice have also indicated the presence of additional tumor suppressor genes for lung cancer. Further analyses of these loci should elucidate how many tumor suppressor genes are involved in human lung carcinogenesis. Molecular and functional analyses of those genes will make it possible to fully understand the molecular mechanism of lung carcinogenesis.

Acid Anhydride Hydrolases↗

PubMed1999

Assessment of mucosal immune response in genitourinary tract using urine.

A novel method to assess mucosal immune response in the genitourinary mucosa after immunization with a mucosal vaccine has been developed. In this method, secretory IgA antibody is measured by a highly sensitive enzyme immunoassay (immune-complex transfer enzyme immunoassay) using urine as a specimen. The urinary IgA antibody response could be detected by the immune-complex transfer enzyme immunoassay. In contrast, a conventional enzyme immunoassay (enzyme-linked immunosorbent assay (ELISA)) could not detect this response because of its low sensitivity. Because urine samples can be collected easily and nontraumatically, not only from experimental animals but also from humans, both males and females, the present method may be applicable for assessing the protective efficacy of candidates for mucosal vaccines against sexually transmitted microorganisms, such as human immunodeficiency virus. Furthermore, the usefulness of this method for novel mucosal vaccine formulae was shown for a model in which vaccine antigen and Bordetella pertussis adjuvant were adsorbed onto CaCO, and enclosed in enteric coated capsules.

Adjuvants, Immunologic↗

PubMed1999

Fine structure and morphogenesis of Borna disease virus.

Borna disease virus (BDV), a negative nonsegmented single-stranded RNA virus, has not been fully characterized morphologically. Here we present what is to our knowledge the first data on the fine ultrastructure and morphogenesis of BDV. The supernatant of MDCK cells persistently infected with BDV treated with n-butyrate contained many virus-like particles and more BDV-specific RNA than that of untreated samples. The particles were spherical, enveloped, and approximately 130 nm in diameter; had spikes 7 nm in length; and reacted with BDV p40 antibody. A thin nucleocapsid, 4 nm in width, was present peripherally in contrast to the thick nucleocapsid of hemagglutinating virus of Japan. The BDV particles reproduced by budding on the cell surface.

Animals↗

PubMed1999

Subtraction ICG angiography in Harada's disease.

BACKGROUND/AIM: The significance of indocyanine green (ICG) angiography (ICGA) in Harada's disease still awaits clarification in many respects. This study investigates the details of choroidal lesions observed in Harada's disease by the subtraction method. METHODS: Eight patients with Harada's disease were followed with ICGA. ICG angiograms were obtained with a Topcon high resolution digital fundus camera and processed with a Topcon IMAGEnet computer system. Image subtraction was conducted for analysing serial angiograms taken at about 2 second intervals during the dye transit phase and those taken in the early and middle phases of angiography. RESULTS: Standard ICG images of acute stage disease showed delayed choroidal filling in the early phase. Mid phase angiograms showed areas with bright fluorescence of variable intensity, indicating intrachoroidal ICG leakage. With image subtraction of angiograms with an interval of seconds the choroidal vessels could be imaged sequentially, with the choroidal arteries visualised first, followed by the definition of the choriocapillaris and then the choroidal veins. The choroidal veins with delayed filling were visualised as positive images in serial subtraction angiograms. Subtraction with an interval of minutes showed uneven background fluorescence and bright fluorescence corresponding to the areas of intrachoroidal ICG leakage. After the disease subsided with steroid therapy, angiography revealed an improvement in delayed choroidal filling. Image subtraction by the second allowed a clear visualisation of improved choroidal venous filling, while subtraction by the minute showed homogeneous background fluorescence, eliminating brighter areas. CONCLUSION: Subtraction ICGA demonstrated that delayed filling of the choroidal veins of varying severity occurs in association with hyperpermeability of the choroidal vessels in the course of Harada's disease.

Adult↗

PubMed1999

Peripheral choriovitreal neovascularization in proliferative diabetic retinopathy: histopathologic and ultrastructural study.

We describe the histopathologic and ultrastructural evidence of choriovitreal neovascularization in the peripheral fundus of a non-vitrectomized eye with proliferative diabetic retinopathy (PDR). One eye with PDR was surgically enucleated because of neovascular glaucoma and studied with light and electron microscopy. The eye had neovascular membranes at the ora serrata of the peripheral fundus. The newly formed vessels originated from the choroid, passed through Bruch's membrane and the retina, and extended into the vitreous. These vessels had either developing or mature characteristics. The endothelial cells of the developing vessels contained a bulky cytoplasm with many intracytoplasmic filaments, ribosomes and rough endoplasmic reticulum. Budding endothelial cells were frequently found in the developing vessels. The endothelial cells of the mature vessels had attenuated cytoplasm and fenestrations with diaphragms. These observations suggest that choriovitreal neovascularization in the peripheral fundus is one of the features of PDR.

Choroid↗

PubMed1999

Comprehensive analysis of p53 gene mutation characteristics in lung carcinoma with special reference to histological subtypes.

To date, the characteristics of p53 gene mutations in lung cancer have been extensively investigated. However, current estimates of p53 alterations are inaccurate, since most investigators have limited their analyses to exons 5 to 8 of the p53 gene. We examined 52 lung carcinoma cell lines and 106 primary non-small cell lung carcinomas (NSCLC) for mutations in the entire coding region of the p53 gene, from exons 2 to 11. High resolution single strand conformation polymorphism analysis was performed using a modified electrophoretic apparatus with a high concentration gel (14%) and accurate temperature control. The prevalence of mutations was high (more than 80%) in both small cell lung carcinoma (SCLC) (15 of 18) and NSCLC cell lines (28 of 34), and 9 of 45 mutations (20%) were detected outside the region of exons 5 to 8. The frequency of the mutations in primary NSCLC was 48% (51 of 106) and was significantly different (p=0.01) between adenocarcinoma (39%) and squamous cell carcinoma (67%). A-->G transitions (14%, 6 of 43 cases) as well as G-->T transversions (26%, 11 of 43 cases) were frequently detected with significant strand bias in smoking patients, suggesting that carcinogens causing these mutations are involved in smoking associated lung carcinogenesis.

Adenocarcinoma↗