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Biomedical subjects

K Kitano

Publications and source records attributed to K Kitano.

At least 145 records · Page 8Linked to original sources

Effective production of a human monoclonal antibody against tetanus toxoid by selection of high productivity clones of a heterohybridoma.

A mouse.human-human heterohybridoma, N12-16.63, has been described which produces an anti-tetanus toxoid human monoclonal antibody (MoAb). A clone, N12-16.63.49.19, which produces eight times as much MoAb as that produced by the original cell line, was selected by repeating the recloning and selection twice. Two clones, N12-16.63.49.19.69 and N12-16.63.49.19.127, further selected from this clone produced almost 20 times more than that produced by the original cell line. Though the production of MoAb by these clones gradually decreased with repeating transfers, they still produced a large amount of human MoAb even after 3 months of transfer. Human MoAb (IgM) was isolated from the culture supernatants of the original and high productivity clones and the products were confirmed to be identical. Human MoAb was effectively produced by batch culture on the 20 liter scale or a perfusion culture on the 1 liter scale using these high productivity clones.

Animals↗

Cloning of cDNA encoding a new peptide C-terminal alpha-amidating enzyme having a putative membrane-spanning domain from Xenopus laevis skin.

A cDNA clone encoding a precursor of a peptide C-terminal alpha-amidating enzyme (AE-I) from Xenopus laevis skin was recently isolated and sequenced in our laboratory. In this study, by using the restriction fragment of this clone as a hybridization probe, we have identified the cDNA encoding another new peptide C-terminal alpha-amidating enzyme (tentatively named AE-II) distinct from AE-I. The cDNA encodes a polypeptide of 875 amino acid residues, which contains a region extensively homologous to AE-I precursor at N-terminus. The encoded protein characteristically has a putative membrane-spanning domain near C-terminus. Our results indicate that C-terminal alpha-amide formation of peptides in Xenopus skin is regulated by at least two distinct alpha-amidating enzymes.

Amino Acid Sequence↗

Retinal toxicity of antibiotics: evaluation by electroretinogram.

Toxicity of an intravitreal injection of gentamicin sulfate, disodium sulbenicillin and cefazolin sodium on the retina was investigated by electroretinogram in albino and pigmented rabbits. Recordings were made before injection and 2 hours and 3, 7, 14, and 21 days after injection. Significant differences were found in the susceptibility of the electroretinogram components to various antibiotics as follows. Gentamicin 0.24 mg/0.1 ml irreversibly abolished all the components examined. Sulbenicillin 4.0, 8.0, or 12 mg/0.1 ml transiently suppressed the b-wave and the oscillatory potentials incrementally with increasing dose. Cefazolin 0.5, 2.0, or 5.0 mg/0.1 ml selectively reduced the oscillatory potentials, leaving the a- and b-waves almost unattenuated. The cefazolin-suppressed oscillatory potentials recovered within 14 days after injection. Judging from the most susceptible electroretinogram components to each antibiotic, we recommend intravitreal doses of these antibiotics for clinical use as follows: gentamicin 0.1 mg/0.1 ml, sulbenicillin 2 mg/0.1 ml, and cefazolin 0.25 mg/0.1 ml.

Animals↗

Congenital contracture of the infraspinous muscle. A case report.

Posterior subluxation of the glenohumeral joint and a posterior, winged scapular deformity have been reported in a patient with congenital contracture of the infraspinous muscle. The present case was successfully treated by resection of the contracted band within the infraspinous muscle, glenoid osteotomy, reverse Putti-Platt, and resection of the protruding part of the scapula. To our knowledge, this is the first case reported of congenital contracture of the infraspinous muscle. The pathogenesis of the deformity in this case is discussed.

Adolescent↗

Clinical and manometric evaluation of anorectal function following low anterior resection with low anastomotic line using an EEA stapler for rectal cancer.

Anorectal function was evaluated in eight patients who had low anterior resection of the rectum with a low anastomotic line, using an EEA stapler, with determination of function based on periodic manometric studies and clinical symptoms. Immediately following surgery all patients suffered from frequent bowel actions and soiling. These symptoms improved with time and most patients could enjoy almost normal daily life by the sixth postoperative month. One month after surgery, anal canal resting pressure and maximum squeeze pressure were significantly reduced and rectoanal inhibitory reflex was absent; neither showed a distinct tendency to improve thereafter. Rectal sensation and reservoir capacity, which also were seriously impaired, recovered satisfactorily by the time of the six-month examination. This suggests that an improvement of clinical symptoms following this operation is dependent upon the recovery of reservoir capacity and sensation of the neorectum, and that this operative procedure is a functionally acceptable option for low rectal cancer.

Adult↗

Proliferative effect of human granulocyte colony-stimulating factor on blast cells of acute promyelocytic leukemia.

The effect of human granulocyte colony-stimulating factor (G-CSF) on leukemic cells of acute promyelocytic leukemia (APL) was examined. Mononuclear cells obtained from bone marrow cells containing more than 90% blasts from seven APL patients were incubated in the presence of G-CSF using semisolid and liquid culture systems. On day 7, the cells from all the patients produced many clusters consisting of 8-40 cells. These cells appeared to be promyelocyte-like blast cells in four patients and had differentiated to more mature neutrophils in three patients. On day 14, the number of clusters decreased except for two patients. Blast cells from the two patients showing the increase of blast clusters could proliferate in a liquid culture containing G-CSF. Blast cells cultured for 14 days formed many secondary cultures after replating on a methylcellulose medium. Moreover, chromosomal analyses of blasts cultivated in the presence of G-CSF for 7 days showed t(15;17) in all metaphases in one patient. It appears that the leukemic cells from APL patients could proliferate in the presence of G-CSF.

Adult↗

Confirmation of bone marrow engraftment by detection of M/N blood group antigens on erythroblasts in erythroid bursts.

In order to confirm erythroid engraftment, we examined the changes in blood group antigens expressed on erythroblasts in the haematopoietic colonies of a bone marrow transplant recipient. A 44-year-old female with acute myelogenous leukaemia underwent an allogeneic bone marrow transplantation from her sister donor. Prior to the transplantation, the blood groups of both recipient and donor were analysed, and their M/N groups found to differ, the former being MN and the latter N. Bone marrow mononuclear cells were obtained from the patient on day 21 after bone marrow transplantation and cultured in semi-solid medium. Erythroblasts were collected from the erythroid bursts that had formed, and were subjected to flow cytometry using monoclonal anti-M and anti-N. Anti-N reactive cells accounted for 99.2% of those in the erythroid bursts, while only 0.3% of these cells were anti-M reactive. MN type erythrocytes in the recipient's peripheral blood had been replaced by N type and only 2.0% of erythrocytes were anti-M reactive on the 70th day after BMT. These erythroblasts and erythrocytes were phenotypically N, and originated from the donor haemopoietic stem cells with the success of bone marrow engraftment. From our findings, it appears that the M/N blood group antigens were produced by the erythroid cells themselves.

Adult↗

Difference of cell lineage expression of haematopoietic progenitor cells in Philadelphia-positive acute lymphoblastic leukaemia and chronic myelogenous leukaemia.

It is still difficult to clinically distinguish Philadelphia (Ph1)-positive acute lymphoblastic leukaemia (ALL) from Ph1-positive chronic myelogenous leukaemia (CML) in lymphoid crisis. In this study we tried to discriminate between these two disorders by simultaneous analyses of cell morphology and karyotype in single in vitro colonies. We studied three patients with Ph1-positive ALL and four with Ph1-positive CML in various phases of the disease. Bone marrow and peripheral blood cells obtained directly from all seven patients showed abnormal karyotypes including Ph1-chromosomes. Normal karyotypes were found in a small proportion of cells from two ALL patients, but none were found in any from the CML patients. The patients' mononuclear cells (MNCs) were plated at 1-5 x 10(4)/ml in semi-solid medium containing methylcellulose plus phytohaemagglutinin-stimulated leucocyte conditioned medium and erythropoietin. After 9-14 d cultivation, granulocyte-macrophage, erythroid and mixed colonies obtained were used for simultaneous analysis of cell morphology and karyotype. Morphological examination showed that these colonies contained neutrophils, eosinophils, basophils, macrophages and/or erythroblasts in various combinations. No lymphoblast colonies were obtained under the culture conditions used. Cytogenetic examination revealed that all metaphase cells observed in colonies obtained from Ph1-positive ALL patient MNCs had a normal karyotype, whereas those in colonies from Ph1-positive CML patient MNCs had abnormal karyotypes, including Ph1 chromosomes, suggesting that the difference between the two disorders involved a difference in cell lineage. Our results showed that this method was a practicable method for distinguishing Ph1-positive ALL from Ph1-positive CML in lymphoid crisis.

Adolescent↗

The total heart beats per 24 hours by ambulatory electrocardiography and the changes of heart rate by treadmill exercise test in sick sinus syndrome.

To study the clinical implications of the total number of heart beats per 24 hours (THB), 24 hour ambulatory electrocardiography and treadmill test were performed by sixty patients with sick sinus syndrome (SSS, 58 +/- 12 years old) who underwent overdrive suppression test. Results were compared with thirty control subjects (58 +/- 12 years old). The THB was 74 +/- 11 thousand beats in the SSS group and 99 +/- 10 thousand beats in the control group. The THB and the maximal heart rate (MHR) achieved during the treadmill test were significantly lower in the SSS group than in the control group. However, the exercise duration in patients with SSS was similar to that of the control subjects. The exercise duration and the MHR were correlated to age, but not to the THB in the patients with SSS. There was no significant relationship between the total heart beats per 24 hours and the maximal sinus node recovery time (max. SNRT). We conclude that the THB, independent of the max. SNRT, can be a useful index in diagnosing and assessing the quantity of bradycardia in patients with SSS. The tolerance of exercise and the MHR were not correlated with the THB, in patients with SSS.

Adult↗

Karyotype evolution and multilineage involvement of Philadelphia chromosome-positive clones in blastic transformation of two patients with chronic myelocytic leukemia.

Simultaneous analysis of the cell morphology and karyotypes on single colonies was carried out in two patients with Philadelphia chromosome (Ph1)-positive chronic myelocytic leukemia in blastic transformation in order to clarify the origin of leukemic cells involved. Patient no. 1 was in a typical myeloblastic transformation and patient no. 2 in "basophilic transformation." Both patients exhibited karyotype evolution in blastic phase (BP), so that we could differentiate BP clones with additional chromosomal abnormalities from chronic phase (CP) clones with only Ph1 among single colonies. The number of single colonies yielding two or more analyzable metaphases was 18 in patient no. 1, and 19 in patient no. 2. Among these colonies, only three in patient no. 1 and none in patient no. 2 were from CP clones and 15 in patient no. 1 and 19 in patient no. 2 were from BP clones. Morphological examination revealed that not only blasts but also mature neutrophils, eosinophils, basophils, macrophages, and erythroblasts were derived from BP clones. These results suggested that (1) BP clones developed at the pluripotent stem cell level, (2) additional chromosomal abnormalities were not restricted to occur in a specific cell line representative in BP; and (3) BP clones, if not all, may retain capacity for maturation and differentiation.

Blast Crisis↗

Base sequence-specific interactions of operator DNA fragments with the lambda-cro repressor coupled with changes in their conformations.

The mechanism of interaction of the operator DNA with the lambda-cro repressor protein was investigated using proton n.m.r. and photo CIDNP. Three kinds of DNA duplexes, the lambda-OR3 17-mer, phi80-OR2 19-mer and CRP binding site 22-mer, were prepared, and all of their imino proton resonances of the complexes with lambda-cro were assigned to individual base pairs. By monitoring the assigned signals of the DNA fragments and lambda-cro, it was found that in the complex of lambda-cro with lambda-OR3, two subunits of the cro dimer bind to the right and left halves of the OR3, respectively, and the bidentate binding induces a structural distortion in the middle of the 17-mer. lambda-cro itself also undergoes a conformational change including loosening of the dimeric form. In the complex of lambda-cro with phi 80-OR2, which has a 6-bp sequence common to that of lambda-OR3, one subunit of the cro dimer seems to bind specifically to the common part. However, there is only a slight conformational change in the cro dimer. In the mixture of the CRP binding site 22-mer and lambda-cro, soft contact without any conformational change was observed between them.

Base Sequence↗

A new treatment of painful amputation neuroma: a preliminary report.

Two cases of painful amputation neuroma treated by a new method are described. A vascularized island transfer from a less important finger was used, but the accompanying nerve was divided and its distal end sutured to the proximal end of the nerve in the recipient finger, after excision of the neuroma. Four years and two years later, no clinical evidence of neuroma was present at either amputation stump or donor site.

Amputation Stumps↗

Focal cervical poliopathy causing juvenile muscular atrophy of distal upper extremity: a pathological study.

A new clinical entity under the name of "juvenile muscular atrophy of unilateral upper extremity" was first described in 1959. Although about 150 cases in Japan, and several additional cases in other countries, have been clinically reported in the literature, the pathology has remained unknown because of the benign course of the disease. The first necropsy findings are reported, obtained from a patient with this disease, who died of lung cancer at the age of 38, 23 years after the onset of the disease. The lesions existed only in the anterior horns of the spinal cord at C5 approximately T1, particularly marked at C7 and C8, showing shrinkage and necrosis, degeneration of various degrees of large and small nerve cells, and mild gliosis. The pathological findings differ from those of reported cases of spinal vascular disorders, but some circulatory insufficiency in the territory of the spinal cord would seem to be suggested, although the underlying aetiology remains unknown.

Adult↗

Complete complementary DNA-derived amino acid sequence of canine cardiac phospholamban.

Complementary DNA (cDNA) clones specific for phospholamban of sarcoplasmic reticulum membranes have been isolated from a canine cardiac cDNA library. The amino acid sequence deduced from the cDNA sequence indicates that phospholamban consists of 52 amino acid residues and lacks an amino-terminal signal sequence. The protein has an inferred mol wt 6,080 that is in agreement with its apparent monomeric mol wt 6,000, estimated previously by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Phospholamban contains two distinct domains, a hydrophilic region at the amino terminus (domain I) and a hydrophobic region at the carboxy terminus (domain II). We propose that domain I is localized at the cytoplasmic surface and offers phosphorylatable sites whereas domain II is anchored into the sarcoplasmic reticulum membrane.

Amino Acid Sequence↗

Effects of combined changes in serum calcium and potassium on QT interval. A study by Holter electrocardiographic monitoring during hemodialysis.

Holter electrocardiographic recordings were performed for 24 hours in 20 patients on chronic hemodialysis to evaluate the effect of changes in serum calcium and potassium during a 5-hour hemodialysis period on the QT interval. Hemodialysis caused an acute increase in serum calcium from a predialysis value of 9.6 +/- 1.3 mg/dl to 11.5 +/- 1.2 mg/dl and a decrease in serum potassium from 4.9 +/- 0.6 mEq/L to 3.5 +/- 0.4 mEq/L. The Q-oTc interval shortened from a predialysis value of 0.240 +/- 0.023 sec to a minimum of 0.215 +/- 0.022 sec during the 5th hour of hemodialysis. This Q-oTc interval shortening was correlated strongly with an increase in serum calcium and was statistically significant only in subjects showing an increase in serum calcium of more than 2 mg/dl (p less than 0.05). The Q-eTc interval increased from a predialysis value of 0.392 +/- 0.030 sec to 0.414 +/- 0.024 sec during the 4th hour of hemodialysis. This Q-eTc interval prolongation was correlated with a decrease in serum potassium. It was concluded that increased serum calcium during hemodialysis caused shortening of the Q-oTc interval, that the degree of shortening was proportional to the magnitude of increase in serum calcium, and that changes in serum calcium and the Q-oTc interval were concurrent. It was also concluded that hemodialysis resulted in prolongation of the Q-eTc interval. This change was possibly caused by a decrease in the level of serum potassium during the procedure.

Adult↗