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Biomedical subjects

K Kino

Publications and source records attributed to K Kino.

At least 73 records · Page 4Linked to original sources

Production of chicken chimeras from injection of frozen-thawed blastodermal cells.

To execute a strategy for reconstituting genetic resources from cryopreserved blastodermal cells, experiments were conducted to optimize conditions for producing chimeric chickens from frozen-thawed blastodermal cells. Stage X blastodermal cells were collected from Barred Plymouth Rock embryos and dispersed. Cells were resuspended in 10% dimethyl sulfoxide in Dulbecco's modified Eagle's medium (DMEM) containing 20% fetal bovine serum, and distributed into plastic ampules. Cell suspensions were seeded to induce ice formation at -7 C, cooled from -7 to -35 C at 1 C/min and then ampules were plunged into liquid nitrogen. Thawing was done by plunging the ampules into warm water (37 C) for 3 min. After centrifugation, the supernatant was replaced with DMEM, and dead or broken cells were removed by density gradient centrifugation. Approximately 500 cells were injected into irradiated Stage X White Leghorn recipient embryos. Following incubation, several somatic chimeras were produced. The frequency of somatic chimerism when fresh (unfrozen) cells, or cells that were frozen and selected by density gradient centrifugation on Percoll or Nycoprep were injected into recipient embryos was 84, 79, and 85%, respectively. The percentage of donor-derived pigmentation in the down of these chimeric chickens was 79, 50, and 58%, respectively. Germline chimerism was determined by mating the chimeras that survived to sexual maturity to Barred Plymouth Rocks. Nine of 16 birds (56.2%) injected with fresh cells, 2 of 26 birds (7.7%) injected with cells that were frozen and selected by density gradient centrifugation on a Percoll gradient, and 3 of 26 birds (11.5%) injected with cells that were frozen and selected on a Nycoprep gradient showed germline transmission; the percentage of donor-derived progeny in these chimeras were 29.5, 5.2, and 6.8%, respectively. The Barred Plymouth Rock donor stock was "reconstituted" by inter se mating of germline male and female chimeras. These data demonstrate that the strategy described here for reconstituting genetic resources from cryopreserved blastodermal cells via chimeric intermediates can be performed successfully.

Animals↗

Manipulation of blastodermal cells.

Blastodermal cells isolated from newly laid, unincubated eggs are virtually uncommitted cells that exhibit many of the properties of pluripotential stem cells. They can be transferred from donor to recipient embryos and contribute to both somatic tissues and the germline. Blastodermal cells that have been maintained in culture for 7 d express the epitopes ECMA-7 and SSEA-1, which are also expressed by mouse embryonic stem cells. After culture for up to at least 7 d, blastodermal cells retain the ability to differentiate into somatic tissues and the germline both in vivo and in vitro. Proliferation in the absence of differentiation of blastodermal cells is stimulated by the presence of Leukemia Inhibitory Factor (LIF) and other ligands that interact with the gp130 receptor, and differentiation is stimulated by exposure to retinoic acid. Blastodermal cells also possess high levels of telomerase activity, which is shared by immortalized cells and cells within the germline. Blastodermal cells can be transfected and will express foreign genes both in vivo and in vitro. Transfected cells can be isolated by fluorescence activated cell sorting and can be cryopreserved without losing their ability to contribute to either somatic tissues or the germline. These properties of blastodermal cells make them ideal vectors for introducing genetic modifications to the germline.

Alkaline Phosphatase↗

Hyperproduction of L-threonine by an Escherichia coli mutant with impaired L-threonine uptake.

An efficient production strain for L-threonine fermentation was derived from Escherichia coli by multiple rounds of mutation programs that aimed at deregulation of the L-threonine biosynthetic pathway and blocking of L-threonine degradation pathways. When the optimum amount of DL-methionine was added, this strain KY10935, an L-methionine auxotroph, gave 100 g/liter L-threonine after 77 h cultivation. In this strain, key enzymes in the L-threonine biosynthetic pathway were highly derepressed, but some were inhibited by lower concentrations of L-threonine than the accumulated level. Such incomplete deregulation of the pathway was accounted for by the intracellular concentration of L-threonine being lower than the extracellular level. In an assessment of L-threonine transport in terms of phenotypic growth responses to the amino acid, L-threonine-auxotrophic mutants with a lesion in the L-threonine operon were derived from strain KY10935 by selection for auxotrophy for dipeptide L-alanyl-L-threonine or glycyl-L-threonine, the transport systems of which were different from those of L-threonine. All three independent mutants isolated needed an extraordinarily high concentration (10 mg/ml) of L-threonine, but grew in the presence of a low concentration (10 micrograms/ml) of either dipeptide, indicating that strain KY10935 had impaired L-threonine uptake. These results suggested that the strain had an unusual mechanism of L-threonine hyperproduction: the inability to take up L-threonine that had accumulated extracellularly decreased the steady-state level of intracellular L-threonine, freeing the remaining regulatory steps of feedback inhibition.

Amino Acids↗

[Malignant tumors in a Japanese geriatric hospital].

We studied the cases of 909 patients with malignant tumors (497 men and 412 women) who were admitted to Tokyo Metropolitan Geriatric Hospital from April 1994 to July 1995. The mean age was 78.6 years (range: 60 to 103 years). The most common tumors were gastric cancer, colo-rectal cancer, and lung cancer. However, the most common tumors in those who died (n = 263) were gastric cancer, lung cancer, and liver cancer. In 425, abnormalities were found during routine health checks or incidental laboratory examinations, while the patients were asymptomatic. On the first admission, 23.4% were found distant metastases. The rate of complications with other diseases was 82.3%. About one third (31.2%) were informed of their situation, and 18.5% had not received any treatment for their malignancies. In 63%, performance status at the time of discharge was the same as that measured on admission or better.

Aged↗

[Gastric cancer in the elderly--relationship between macroscopic appearance and histological type in autopsy specimens].

OBJECTIVE: We sought to clarify the characteristics of gastric cancer in the elderly. METHODS: We examined 349 specimens of gastric cancers. All specimens were obtained at autopsy from patients who were over 65 years old. We analyzed the relations among five factors: age of the patient, histological type, macroscopic appearance, grade of invasion and location. RESULTS: Older patients were more likely to have papillary adenocarcinoma, tubular adenocarcinoma, and Borrmann-2 and 3 lesions. Papillary and tubular adenocarcinoma together accounted for 14 of 84 Borrmann-4 lesions (16.3%). Borrmann-1 signet-ring-cell carcinomas were observed in two cases. Tubular adenocarcinomas made up 31 in 38 depressed-type early carcinomas (81.6%). Over half of the papillary and tubular adenocarcinomas were confined to the mucosal or submucosal layers, but over 60% of the poorly differentiated adenocarcinomas or signet-ring-cell carcinomas had invaded the extraserosal layer. Papillary and tubular adenocarcinomas were commonly found in the lower part of the stomach, but poorly differentiated adenocarcinomas and signet-ring-cell carcinomas were found in all parts of the stomach. CONCLUSIONS: Unlike specimens obtained during surgery, many of those obtained at autopsy were from older patients, and the gastric cancers were advanced. Compared to many reports about specimens obtained during surgery, tubular or papillary adenocarcinomas are not a few histological type in Borrmann-4 lesions in this study. Surgical specimens of depressed-type early cancer in younger patients are most commonly found to be signet-ring-cell carcinoma, but tubular adenocarcinoma was common in this study. We found many differences between specimens obtained at autopsy and during surgery. These findings indicate that autopsy studies can reveal some otherwise-obscure characteristics of gastric cancer in the elderly.

Adenocarcinoma↗

[Coronary artery bypass grafting using arterial grafts in a patient with rheumatoid arthritis requiring steroid therapy].

We report a 58-year-old man with rheumatoid arthritis (RA) who underwent successful coronary artery bypass grafting (CABG) for stable angina after myocardial infarction. He had been receiving prednisolone for 9 years. Selective coronary angiography revealed 99% stenosis in the left anterior descending branch (LAD; seg. 6). Before going on cardiopulmonary bypass, the inferior epigastric artery (IEA) was anastomosed to the left internal thoracic artery (LITA) in end-to-side fashion. The LITA was anastomosed to LAD (seg. 7) and the IEA was anastomosed to the first diagonal branch (seg. 9). The postoperative course was uneventful and postoperative angiography showed both grafts were well patent. He is now doing well with no angina attack. The stumps of the LITA and IEA were examined pathologically with the atherosclerotic changes of both grafts relatively mild. We consider the arterial grafts are useful for CABG even in the patient with RA requiring steroid therapy.

Angina Pectoris↗

[Successful open commissurotomy and resection of dysplastic myxomatous tissue on the leaflet edges in a neonate and an early infant with critical aortic stenosis].

A neonate and an early infant with critical aortic stenosis successfully underwent open commissurotomy and resection of dysplastic myxomatous tissue on the leaflet edges using cardiopulmonary bypass. Case 1: A 31-day-old boy admitted to our unit with shock. Echocardiography demonstrated critical aortic stenosis and severe left ventricular dysfunction (EF = 15%). Case 2: A 12-day-old boy suddenly deteriorated and required resuscitation with ventilation and inotropic support. Emergency operation was required in both cases using cardiopulmonary bypass with systemic hypothermia (30 degrees C). In both cases, the aortic valve was bicuspid and dysplastic with gelatinous myxomatous tissue on the leaflet edges. Commissurotomy and resection of myxomatous tissue were performed. Their postoperation courses were uneventful and they have been free from medication at present. These results suggest that aortic commissurotomy and resection of myxomatous tissue under direct vision may be preferable for critical aortic stenosis with dysplastic aortic valve.

Aortic Valve↗

[Transverse submammary skin incision for congenital cardiac surgery].

Submammary skin incision has been accepted as cosmetic approach for open heart surgery. During November 1990 to February 1995, 39 ASD patients under 15 years old were operated with either median or submammary skin incision in Okayama University Medical School. For these patients, retrospective study was carried out to determine whether submammary skin incision made surgical procedure complex and whether patients were satisfied with the scar by submammary skin incision. There were 14 submammary skin incision (Group S) and 25 median skin incision (Group M). There was no significant difference in patients profile between the two groups except for sex. Operative time, cardiopulmonary bypass time and aortic cross clamp time had no significant difference between the two groups. Postoperative ICU stay and hospital stay also did not differed between the two groups. Questionnaires for evaluation of the subjective operation scar was sent with 32 answers (82%). There was no significant difference on questionnaire findings. However for the patients without hypertrophic scar, more patients in Group S were significantly satisfied with their scars. Submammary skin incision was safe and easy. It could be alternative of median skin incision. If hypertrophic scar formation could be prevented, more patients would be satisfied with submammary skin incision than median skin incision.

Cardiac Surgical Procedures↗

Aureobasidin A, an antifungal cyclic depsipeptide antibiotic, is a substrate for both human MDR1 and MDR2/P-glycoproteins.

The human MDR1 gene encodes the multidrug transporter P-glycoprotein (Pgp). Although the MDR2/Pgp shares about 80% identity at the amino acid level with the MDR1/Pgp, the MDR2/Pgp cannot act as a multidrug transporter. We examined the drug sensitivity of Saccharomyces cerevisiae expressing either the human MDR1/Pgp or MDR2/ Pgp. The human MDR1/Pgp conferred about 4-fold resistance to aureobasidin A, a cyclic depsipeptide antifungal antibiotic, on the drug-sensitive yeast strains. Interestingly the human MDR2/Pgp also conferred about 2.5-fold resistance to aureobasidin A. The resistance to aureobasidin A conferred by the MDR2/Pgp as well as by the MDR1/Pgp was overcome by vinblastine, verapamil, and cyclosporin A, depending on their concentrations, but not by colchicine. Aureobasidin A probably interacts directly with Pgps, because it overcame multidrug resistance of human cells and inhibited azidopine photoaffinity labeling of MDR1/Pgp in human cell membranes. These results suggest the possibility that the human MDR1 and MDR2/Pgps have conserved domain(s) for drug recognition.

ATP Binding Cassette Transporter, Subfamily B↗

Establishment and characterization of a human colon cancer cell line, OUMS-23, from a patient with familial adenomatous polyposis.

A human colon carcinoma cell line designated OUMS-23 has been established from metastatic pericardial fluid of a male familial adenomatous polyposis patient with colon cancer. Since 1984, the epithelial cells have been maintained in culture. Ultrastructural studies revealed the presence of numerous microvilli on the cell surface and desmosomes between the adjacent cells. The cells secreted carcinoembryonic antigen into the culture medium (15 ng/10(6) cells-1 24 h-1). The cells expressed heat-stable placental-type-like alkaline phosphatase, whereas the normal counterparts expressed tissue-unspecific alkaline phosphatase. Karyotypic analysis showed that the cell line was of human origin and that the chromosome number was broadly distributed between 53 and 118. Southern blot analysis of the APC gene revealed no abnormalities in OUMS-24 cells, while Northern blot analysis demonstrated that the expression of the gene was about one-half that of the normal human fibroblasts. No mutations at the "hot spots" of codons 12 and 61 of H-, K- and N-ras proto-oncogenes were detected in the cells. The cells could grow in soft agar at a cloning efficiency of 6.5%, and upon transplantation into nude mice the cells formed tumors, which were diagnosed as differentiated adenocarcinoma.

Adenomatous Polyposis Coli↗

Late aneurysm after subclavian flap aortoplasty for coarctation of the aorta.

Late aneurysms are common after repair of coarctation of the aorta by prosthetic patch aortoplasty but are rare after subclavian flap aortoplasty. We present the case of a 14-year-old boy who underwent a grafting procedure for a descending thoracic aortic aneurysm after subclavian flap aortoplasty for coarctation of the aorta when he was 2 years old. This is the fifth report of late aneurysm formation after subclavian flap aortoplasty.

Adolescent↗

rHuEPO enhances the production of plasminogen activator inhibitor-1 in cultured endothelial cells.

The direct effects of recombinant human erythropoietin(rHuEPO) on coagulation and fibrinolysis factors were evaluated in a cultured endothelial cell (EC) system. Confluent quiescent ECs were incubated with or without 5.0 U/ml rHuEPO for 1, 6, and 18 hours, and supernatant concentrations of plasminogen activator inhibitor-1 (PAI-1): antigen (Ag), tissue plasminogen activator and thrombomodulin, and supernatant activities of tissue factor pathway inhibitor and von Willebrand factor were measured. The results showed that only PAI-1 levels were increased by the presence of rHuEPO. In order to assess the effect of rHuEPO on PAI-1 production by EC more precisely, confluent ECs were incubated with various doses of rHuEPO (0, 1.0, 2.5, 5.0, 10.0 U/ml) for 1, 6, 12, and 18 hours, and PAI-1:Ag concentrations in the supernatants of media were measured. PAI-1:Ag in the supernatants were increased by the presence of rHuEPO at all incubation times (P < 0.01) and the increase in PAI-1:Ag was dependent on rHuEPO concentration. The increases in PAI-1:Ag by 5.0 U/ml rHuEPO were comparable to those by 0.1 U/ml tumor necrosis factor-alpha, 1.0 microgram/ml lipopolysaccharide, and 0.5 U/ml thrombin. The increase in PAI-1:Ag by rHuEPO was suppressed by pre-incubation with 10 micrograms/ml cycloheximide (P < 0.01) or 0.2 microgram/ml actinomycin D (P < 0.01). These results indicate that rHuEPO directly stimulates PAI-1 production in cultured EC via de novo protein and RNA syntheses.

Cells, Cultured↗

Epitope analysis of HLA-DR-restricted helper T-cell responses to Der p II, a major allergen molecule of Dermatophagoides pteronyssinus.

T-cell epitopes of Der p II, a major allergen of Dermatophagoides pteronyssinus, were analyzed by using human T-cell clones. We tested 38 cloned T cells from two Japanese patients with allergic rhinitis, and identified at least two peptides (K33-T47 and I58-C73) as helper T-cell epitopes. The former epitope was shown to be restricted by HLA-DRB1*1502, and the latter by HLA-DRB1*0405, both of which are typical Japanese HLA-DR alleles, suggesting that those T-cell epitopes might be important for the onset of house-dust mite allergy in the Japanese population. We prepared 15 analog peptides of the HLA- DRB1*1502-restricted 15-mer peptide. Of those 15 residues, five (F35, L37, A39, F41, and E42) were critical for the epitope activity, and three residues (F35, A39, and E42) seemed to be included in anchor motifs for HLA-DRB1*1502. The epitope peptide was also recognized by HLA-DRB1*1502-positive healthy donors; however, only allergic T cells showed Th2 functions. Antigen-presenting cells of nonallergic donors were able to activate allergic T cells to express Th2 function. This seemed to suggest that antigen recognition of T cells, as well as additional unknown factors which promote Th2, rather than Th1, responses, might be important for the onset of house-dust mite allergy.

Allergens↗

Arthrotomographic sign of a 'double line' as an indicator of disc folding and sideways disc displacement in the temporomandibular joint.

Dual-space, single-contrast arthrotomolgraphy of the TMJ was performed on a 19-year-old woman with a history of pain and clicking. Simultaneous multi-layer arthrotomography with computed radiography was used for the present examination. Anterior disc displacement with reduction was observed in the sagittal plane, together with a double line in the lower joint space. The double line was interpreted as disc folding and sideways disc displacement and was confirmed in the coronal plane.

Adult↗

[Clinical and pathological study of gastric cancer treated without surgery in elderly patients].

OBJECTIVE: To clarify the clinical and pathological characteristics of gastric cancer in the elderly. METHOD: Clinical and pathological characteristics of patients who died during 1986-94 with gastric cancer which did not undergo surgery were studied retrospectively. The patients were divided into three groups according to age: A (65-74 years, n = 38), B (75-84 years, n = 77), and C (85 years and over, n = 43). RESULTS: Borrman type-1 was more frequent in group C and Borrman type-4 was frequent in group B. "Early cancer", protruded type (type I or type IIa) was more frequent in group C. In cases of advanced cancer some patients did not undergo surgery because the cancer was to advanced, and others had concomitant diseases that made surgery risky. In all groups, a few patients refused surgery. In cases of "early cancer", the most reason for not undergoing surgery was a concomitant disease in all groups. Other reasons were general weakness and dementia in group C, second malignancy and dementia in group B, and second malignancy in group A. After advanced cancer was diagnosed, 9.6% survived for 1 year, and 1.7% for 2 years. After early cancer was diagnosed, 50.7% survived for 1 year, 15.2% for 3 years and 7.8% for 4 years. The survival rates did not differ by age. In all age groups, gastric cancer was frequently the main cause of death in patients with advanced cancer, but many patients with "early cancer" died of other causes.

Age Factors↗

[Subaortic stenosis].

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Aortic Stenosis, Subvalvular↗