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Biomedical subjects

K Sawada

Publications and source records attributed to K Sawada.

At least 235 records · Page 13Linked to original sources
PubMed1994

Transitional changes in immunophenotypic subpopulations of human peripheral blood CD34+ cells expanded in vitro.

We determined the appropriate incubation period to expand human peripheral blood (PB) CD34+ cells for clinical application and the role of recombinant human (rh) interleukin-3 (rhIL-3) in the expansion and differentiation of these cells. The cells were purified up to 40 +/- 16% and transitional changes in immunophenotypic subpopulations in suspension culture were examined following stimulation with three different combinations of rh colony-stimulating factors (rhCSFs): 1) rhIL-3 alone, 2) rhIL-6, rhSCF, rhG-CSF, plus rhGM-CSF, and 3) the four CSFs plus rhIL-3. With all three CSF combinations, the total cells increased continuously after day 5 until day 14, and a combination of the five CSFs always gave rise to the highest number of total cells. Immunophenotypic analysis of the expanded cells showed that with all three CSF combinations CD34+ cells reached a maximal rate on day 5 and then decreased in an inverse correlation between the logarithm of CD34 positive rate and the duration of suspension culture. The maximum expansion of CD34+ cells and PB progenitor cells (PBPC) with rhIL-3 alone, the four CSFs, or the five CSFs was observed on day 5, 10, or 7, respectively. The combination of the five CSFs was identified as the most potent stimulus for expansion of PBPC and CD34+ cells, as it increased non-erythroid PBPC 89 +/- 69-fold, with a range of 24 to 204-fold on day 7. However, differences in the expansion rate of these cells on days 5, 7, and 10 were not statistically significant. The majority of purified CD34+ cells coexpressed CD38 (91 +/- 3%) but were negative for CD33 (85 +/- 4%), and one-half coexpressed CD13. With all three combinations of CSFs, the mature CD34+ cells that coexpressed CD38, CD33, or CD13 expanded in parallel with the total CD34+ cells, while an increase in relatively immature CD34+ cells, which do not express CD38, CD33, or CD13, was only statistically significant with the five CSFs. Thus, rhIL-3 played a critical role when combined with the four CSFs by increasing both mature and immature CD34+ cells.

Adult↗
PubMed1994

[Chronic cold agglutinin disease accompanied with an increase of CD20+/CD5+ cells; a case report].

A 61-year-old male complained of acrocyanosis and dark urine when exposing to cold temperatures. This had continued for several years. His physical examinations showed neither lymphadenopathy nor hepatosplenomegaly. Laboratory findings were as follows; RBC 305 x 10(4)/microliters, Hb 10.3 g/dl, reticulocytes 4.32%, platelets 27.3 x 10(4)/microliters, WBC 7,400/microliters with 50% lymphocytes, and a high cold agglutinin titer (2,048-fold) with anti-I specificity. Bone marrow smear preparations showed erythroid hyperplasia and increase of lymphocytes (52%). Immunophenotypic analysis showed an increase of CD20+/B-lymphocytes in peripheral blood (32.6%) and in bone marrow, and 94% of these cells co-expressed CD5. Most B-lymphocytes expressed surface IgM-lambda, suggesting a monoclonal proliferation of B-lymphocytes. At this point we diagnosed cold agglutinin disease (CAD) because there was no evidence of lymphoma, and the absolute number of peripheral blood lymphocytes was lower than the criteria of chronic lymphocytic leukemia (CLL) proposed by the International Workshop (1989). However, there still remains the possibility of the transitional form between "idiopathic" CAD and B-CLL or lymphoma.

Anemia, Hemolytic, Autoimmune↗
PubMed1994

[Home chemotherapy for peritoneal carcinomatosis].

Nineteen patients diagnosed with peritoneal carcinomatosis were treated by in-home chemotherapy over a period of four years from August, 1990 to July, 1994. Primary diagnoses of the four male and 15 female patients included 12 cases gastric cancer (four males, eight females), five cases of ovarian cancer, one case (female) of appendicular cancer, and one case (female) of breast cancer. In addition to oral administration of UFT-E and 5'-DFUR, chemotherapy included weekly intravenous injection of a massive dose of 5-FU (1,000 mg/m2), subselective intraaortic infusion and intraperitoneal infusion using a reservoir. These methods were used individually and in combination. The drugs used included 5-FU, CBDCA, CPA, THP, and EPIR. Subselective intraaortic infusion was performed by low dose continuous infusion using the Baxter infusor multiday type. Six gastric cancer patients lived normally for over one year, while four died in less than a year. All ovarian and appendicular cancer patients were CR, the breast cancer patient was PR. Ten patients continued working at their jobs while receiving at home chemotherapy treatments. Diuretics were used to alleviats ascites. Although there were no side effects on digestive organs, 5-FU and CBDCA were mixed with 100 mg hydrocortisone in the infusor to improve cachexia, and promote appetite and activity. Bone marrow suppression was very slight at these dosages, and weekly checkups were adequate. The at-home rate (number of days at home/entire period since onset) of all patients was 78%.

Adult↗
PubMed1994

[Multiple myeloma with massive ascites fluid--immunophenotypic analysis of myeloma cell and its IL-6-dependent growth].

A 75-year-old female was diagnosed as having multiple myeloma (IgG.lambda type. Stage IIA) with plasmacytoma of the head and back in October, 1989. She obtained partial remission by MCNU and MP therapy, but relapsed with massive ascites in January, 1991. VAD therapy was not effective and she died of multiple organ failure on February 23. Her ascites contained a large number of myeloma cells, and the phenotypic analysis and the response to interleukin-6 (IL-6) of these myeloma cells were examined. The myeloma cells were positive for CD33, CD45, CD45RA, CD63, CD71, plasma cell associated antigens such as CD38, PCA-1, BL3, and various kinds of adhesion molecules: CD11a/CD18 (LFA-1), CD29 (VLA-beta 1), CD44 (H-CAM), CD49d (VLA-4), CD54 (ICAM-1), CD56 (N-CAM), CD58 (LFA-3). IL-6 level in the ascites was increased at 91.0pg/ml. The myeloma cells showed an IL-6 dependent growth, which was inhibited by anti-IL-6 antibody (Ab) and anti-IL-6 receptor Ab in vitro. Myeloma cells appearing in ascites have rarely been reported. Our case suggested that IL-6 was a potent growth factor of myeloma cells through an autocrine mechanism in the ascites, and resulted in an aggressive myeloma.

Aged↗
PubMed1994

Simultaneous occurrence of adenomas in stomach, duodenal bulb, and colon: disappearance of gastric and duodenal adenomas during oral administration of 5'-deoxy-5-fluorouridine.

The present report concerns a patient who was first diagnosed as having adenoma in the duodenal bulb and subsequently was discovered to have adenomas in the stomach and colon. Surgical procedures including endoscopic resection could not be carried out because the patient had advanced liver cirrhosis. Follow-up colonoscopy no longer detected any polyp in the colon. Ultrasonography demonstrated a slowly growing hypoechoic mass in the liver, which was indicative of hepatocellular carcinoma. Then, single chemotherapy with oral administration of 5'-deoxy-5-fluorouridine was initiated. Surprisingly, repeated esophagogastroduodenoscopy revealed gradual regression of the gastric and duodenal tumors. Findings from the endoscopy and biopsy materials 10 months after the start of therapy showed disappearance of the gastric and duodenal adenomas. Despite enlargement of the hepatic mass, no recurrence of gastric and duodenal tumors has been noted during the past 1 1/2 yr.

Adenoma↗
PubMed1994

Omega-agatoxin-TK containing D-serine at position 46, but not synthetic omega-[L-Ser46]agatoxin-TK, exerts blockade of P-type calcium channels in cerebellar Purkinje neurons.

omega-Agatoxin-TK (omega-Aga-TK), a 48-amino-acid peptide isolated from the venom of the funnel web spider (Agelenopsis aperta), is a selective and potent inhibitor of P-type calcium channels in the nervous system. We have synthesized a peptide that has the amino acid sequence identified for native omega-Aga-TK. The synthetic omega-Aga-TK, however, showed 80-90-fold less potent inhibition of P-type calcium channels, compared with native omega-Aga-TK. Enantiomer analysis of native omega-Aga-TK revealed D-serine at position 46, and synthetic omega-[D-Ser46]Aga-TK had the same potency as native omega-Aga-TK for blocking P-type calcium channels in cultured cerebellar Purkinje neurons. Two peptide fragments of omega-Aga-TK, namely omega-Aga-TK(1-43) and the carboxyl-terminal peptide fragment omega-Aga-TK(44-48), did not produce any significant inhibition of P-type calcium channels or interfere with the blockade of the channels elicited by native omega-Aga-TK. Molecular dynamics calculations showed that the carboxyl-terminal, six-amino-acid peptide of omega-Aga-TK containing D-Ser46 assumes a different conformation than does the peptide containing L-Ser46. These results suggest that the specific conformation of the carboxyl-terminal region of omega-Aga-TK, particularly the configuration of Ser46, together with a beta-sheet structure formed by four disulfide bonds, might be essential for blockade of P-type calcium channels.

Agatoxins↗
PubMed1994

[A case with AML (M4EO) accompanied by recurrent reactive histiocytosis which showed spontaneous remission].

A 30-year-old female was admitted to our hospital complaining of high fever and fatigue. Laboratory findings showed as follows; WBC 41,500/microliter (40% of blasts), Hb 8.5g/dl, platelets 4.4 x 10(4)/microliter. Cytochemical staining of blasts was positive for peroxidase and non-specific esterase with NaF inhibition. Chromosome analysis showed 46, XX, inv (16p+,q-). AML with eosinophilia was diagnosed. During myelosuppression after remission induction therapy, she developed high fever, and did not respond to transfusions. Marrow smears showed the presence of phagocytic histiocytes consisting of 18% total nuclear cells. A diagnosis of reactive histiocytosis (RH) was made. She recovered spontaneously, but suffered two episode of recurrence during subsequent chemotherapy. Reactive histiocytosis is characterized by proliferation of histiocytes which phagocyte blood cells in immunodeficient cases, e.g. a myelosuppressive state after chemotherapy. RH causes high fever and prolonged myelosuppression. It is considered to be one of the poor prognostic factors in AML during chemotherapy, and spontaneous recovery is rare. In this report, the effect of hydrocortisone on histiocytes derived from patient marrow was also investigated in vitro.

Adult↗
PubMed1993

In vitro expansion of human peripheral blood CD34+ cells.

To elucidate the role of recombinant human colony-stimulating factors (CSFs) for expanding peripheral blood (PB) CD34+ cells, these cells were purified up to 94.5% +/- 1.3% and the effects of individual and combined CSFs on the proliferation and differentiation of these cells were studied in a 7-day suspension culture. The majority of CD34+ cells coexpressed CD38 (81.8% +/- 5.1%), but was negative for CD33 (88.5% +/- 3.4%). Among the individual CSFs examined, recombinant interleukin-3 (rIL-3) was identified as the most potent factor for expanding PB progenitor cells and increased nonerythroid progenitor cells 13- +/- 4-fold (P < .01). Recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF), recombinant granulocyte-CSF (rG-CSF), recombinant macrophage-CSF (rM-CSF), rIL-6, rIL-11, and recombinant stem cell factor (rSCF) did not alone expand nonerythroid progenitor cells. A combination of 5 CSFs, ie, rIL-3, rIL-6, rGM-CSF, rG-CSF, and rSCF, was identified as the most potent combination of those tested and increased nonerythroid progenitor cells 57- +/- 11-fold. After a 7-day suspension culture of CD34+ cells with these 5 CSFs, CD34+ cells expanded 14.5-fold, and CD34+/CD33- cells and CD34+/CD33+ cells were also expanded 2.9-fold and 307-fold, respectively. Most secondary colonies derived from expanded cells were small; however, the absolute number of large-sized colonies expanded 5.9- +/- 3.3-fold. Thus, the combination of CSFs can achieve a degree of amplification of PB CD34+ cells. The capability of in vitro expansion of PB CD34+ cells as an adjunct to PB stem cell transplantation is worthy of consideration.

Adult↗
PubMed1993

A novel peptide from funnel web spider venom, omega-Aga-TK, selectively blocks, P-type calcium channels.

In the course of purification of omega-Aga-IVA, a specific P-type calcium channel blocker, from the venom of Agelenopsis aperta we discovered a novel peptide. This peptide, named omega-agatoxin Tsukuba (omega-Aga-TK), also blocked P-type channels and was twelve times more abundant in the venom than omega-Aga-IVA. omega-Aga-TK was purified to homogeneity by a two-step reverse-phase HPLC procedure. Its amino acid sequence is 71% identical to that of omega-Aga-IVA. omega-Aga-TK has a negatively charged N-terminus, whereas omega-Aga-IVA has a positively charged one. Electrophysiological data indicate that omega-Aga-TK is a potent and selective inhibitor of P-type channels.

Agatoxins↗
PubMed1993

Evaluation of a relationship between polymer bulk hydroxyl and surface oxygen content and in vitro serum-material interaction.

To evaluate serum-material interaction for six types of minimodule and to assess the relationship between the interaction and chemical composition, notably bulk polymer hydroxyl (-OH) percent of polymer, or surface oxygen (SO) percent, polymeric membranes with varying -OH and SO percents were evaluated with normal human serum. The membrane materials (-OH percent and SO percent) evaluated included polypropylene (PP; 0% and 1.9%), polyvinyl alcohol (PVA; 23.7% and 27.3%), ethylene vinyl alcohol (EVAL 4A and D; 30.4% and 25.3%), Cuprophan (CP; 31.5% and 37.4%), and Hemophan (HP; 30.9% and 23.6%), respectively. Data from serum perfusions expressed as percent changes to sham perfusion showed that solute percent decreases were less than 10% in all materials except PVA (10-22%). PVA and CP had higher C3a, C4a, and C5a, and C3a concentration increases, and had larger suppressive effects for all three mitogen-induced mononuclear cell transformation functions (MNCTF) and concanavalin A-induced MNCTF, respectively. PVA had higher SO percent than EVAL and CP was higher than HP, despite PVA and CP having lower or comparable bulk -OH percent to EVAL or HP. The results obtained in the serum material interaction studies related more with the SO percent of the polymer rather than bulk -OH percent. The differences for C4a and PHA-induced MNCTF observed between the two EVAL membranes may be associated with significantly different pore size and therefore different surface structural properties. These results suggest that surface chemical (SO percent on the materials) and structural property analyses are important factors in biocompatibility parameter studies.

Biocompatible Materials↗
PubMed1993

Current status of type 2 (non-insulin-dependent) diabetic subjects on dialysis therapy in Japan.

According to a national survey of dialysis patients in Japan conducted by the Japanese Society for Dialysis Therapy, there were 1,033 patients on dialysis in the Shiga area which has a population of about 1.2 million. Of these 1,033 dialysis patients 140 were the result of diabetic nephropathy. From four hospitals affiliated to Shiga University of Medical Science the medical records of 90 diabetic subjects on dialysis therapy were reviewed and various clinical parameters were analysed and compared with those of patients with chronic glomerulonephritis. Since only one patient had Type 1 (insulin-dependent) diabetes, the remaining 89 with Type 2 (non-insulin-dependent) diabetes were used for this study. The significantly different variables between patients with Type 2 diabetes and chronic glomerulonephritis were age (60.4 vs 54.6 years, p < 0.05), BMI (22.4 vs 20.6 kg/m2, p < 0.001), cardiothoracic ratio (56.4 vs 53.3%, p < 0.001), mean blood pressure (110 vs 117 mmHg, p < 0.05), serum creatinine (9.0 vs 11.5 mg/dl, p < 0.001), serum urea-N (98.2 vs 115.5 mg/dl, p < 0.001), serum total protein (6.0 vs 6.5 g/dl, p < 0.001) and serum albumin (3.5 vs. 3.9 g/dl, p < 0.001). Serum levels of cholesterol and triglyceride were not significantly different between two groups, though the prevalence of electrocardiogram abnormalities, oedema, neuropathy, myocardial infarction and cerebrovascular diseases was significantly higher in the Type 2 diabetic group. These results suggested that Type 2 diabetic patients with end-stage renal disease were older, more malnourished, fluid overloaded and multi-morbid as a result of vasculopathy and neuropathy.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Pressure↗
PubMed1993

A novel stage-specific differentiation antigen is expressed on mouse testicular germ cells during early meiotic prophase.

A murine cell surface antigen exhibiting stage-specific expression during spermatogenesis was detected with two monoclonal antibodies (mAbs), designated BC7 and CA12. In mouse testis, these mAbs recognized a small population of cells located near the periphery of seminiferous tubules at stages XII and I-VI, and these spermatogenic cells were identified as zygotene and early pachytene spermatocytes. Expression of the antigens was transient and was not detected in germ cells at more advanced stages of spermatogenesis such as late pachytene spermatocytes and round spermatids. Immunoprecipitation and immunoblotting studies showed that both mAbs CA12 and BC7 reacted with the same antigenic molecule, which had an estimated molecular mass of 95 kDa. CA12/BC7 antigen, detected in plasma membrane fraction, was a glycoprotein with sialic acid residues and had affinity with WGA lectin. Furthermore, intraperitoneal injection of mAb BC7 caused an apparent spermatogenic disturbance in prepubertal mice. These results suggested that CA12/BC7 antigen, a novel cell surface glycoprotein, is an essential molecule that plays an important role during early meiotic prophase of spermatogenesis.

Animals↗
PubMed1993

Proliferation and differentiation of myelodysplastic CD34+ cells in serum-free medium: response to individual colony-stimulating factors.

The presence of serum or contaminant cells may alter clonal development of haematopoietic progenitor cells in vitro. To investigate the pathogenesis of myelodysplastic syndromes (MDS), marrow progenitor cells from 13 MDS patients were highly purified using monoclonal antibodies including CD34 and immunomagnetic microspheres. The cells positive for CD34 in the purified cells were in the range from 87% to 98%. These purified cells were cultured in serum-free medium with individual colony stimulating factors (CSFs) to investigate whether CD34+ cells from MDS patients have abnormal responses to individual CSFs. Dose response experiments with the purified CD34+ cells and recombinant human macrophage-CSF (rM-CSF), granulocyte-CSF (rG-CSF), granulocyte/macrophage-CSF (rGM-CSF), interleukin-3 (rIL-3) or erythropoietin (rEP) were performed in serum-free fibrin clots in 11 patients. Five patients showed a diminished response to rG-CSF and one patient to rEP. In the remaining six patients the purified CD34+ cells did not respond to a stimulation of any individual CSFs. The results indicate that the progenitor cell growth abnormalities in these disorders involve a defect in the capacity of progenitor cells to respond to stimulation with G-CSF, and present direct evidence for the manner in which myelodysplastic CD34+ cells are impaired.

Adult↗
PubMed1993

Genetic characterization of the multipotent dedifferentiated state of pigmented epithelial cells in vitro.

Retinal pigmented epithelial cells (PECs) of chicken embryos extensively and almost synchronously transdifferentiate into lens cells in medium containing phenylthiourea and testicular hyaluronidase, passing through the bipotent dedifferentiated state. We have isolated genes that are expressed specifically by either pigment or lens cells and analyzed their expression in the transdifferentiation process. The expression of some proto-oncogenes was also studied. In the dedifferentiation process, expression of the c-myc gene was enhanced and the transcription of PEC-specific genes (MMP115, pP344) was completely repressed. However, transcription of lens-specific genes (alpha-, beta- and delta-crystallins genes) remained silent in dedifferentiated pigment cells. Activation of len- or PEC-specific genes occurred only in conditions permissive for lens or PEC differentiation, respectively. These results indicated that lens transdifferentiation from PECs proceeds through a multipotent (or at least bipotent) intermediate cell state in which the c-myc gene is activated, but neither PEC- nor lens-specific genes are expressed.

Animals↗
PubMed1993

Evidence that the chick lens cytoskeletal protein CP 49 belongs to the family of intermediate filament proteins.

A partial cDNA sequence for chick lens beaded-filament protein CP 49 showed the greatest similarity to the sequence of acidic cytokeratins, especially human cytokeratin 18. The predicted amino acid sequence of chick CP 49 corresponded to the entire coil 1a region of the rod domain of human cytokeratin 18, spacer 1, coil 1b, spacer 2 and about half of coil 2. For this sequence of 242 amino acids, there was an overall 38% identity and 76.8% similarity between the chick CP 49 and human cytokeratin 18. This is further evidence that CP 49 belongs to the family of intermediate filament proteins.

Amino Acid Sequence↗
PubMed1993

A set of anti-crystallin monoclonal antibodies for detecting lens specificities: beta-crystallin as a specific marker for detecting lentoidogenesis in cultures of chicken lens epithelial cells.

Seven hybridoma lines which produced monoclonal antibodies against lens crystallins were established. They could detect alpha A-, alpha B-, gamma-, delta-, and several beta-crystallins of different species of animals with high avidities. Although immunohistological analysis of chicken and mouse lenses showed the typical distributions of each crystallin as have been reported so far, the ectopic expression of alpha B or delta-crystallin was observed in the brain or the kidney of chicken, respectively. Subsequently, crystallin production during the process of lentoidogenesis in cultures of chicken lens epithelial cells was examined with these antibodies. Western blot analysis revealed that beta-crystallins were not detected in cultured cells before lentoidogenesis, although all other crystallins were detected throughout the culture period. Immunostaining of cultures indicated clearly that expression of beta-crystallins was restricted to lentoid bodies. These data confirmed the lens fiber specificity of beta-crystallins as previously reported in the messenger RNA level. In addition, we found that small-size delta-crystallin (48 kDa) accumulated before the onset of lentoidogenesis. These results strongly suggest that the differentiated state of lens cells in vitro could be classified by examination of the expression pattern of crystallins. In addition, the anti-crystallin monoclonal antibodies produced in this study could be useful for detecting lens specificities.

Animals↗