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Biomedical subjects

K Ohno

Publications and source records attributed to K Ohno.

At least 199 records · Page 11Linked to original sources
PubMed1999

Comparison of recording systems and analysis methods in specular microscopy.

PURPOSE: To compare corneal endothelial cell images from contact and automated noncontact specular microscopes and to compare endothelial image analysis by the Konan Robo Center Method and the Bio Optics Bambi Corners Method. METHODS: Twenty-six normal corneas of 13 subjects and 41 penetrating keratoplasties (PKs) of 38 patients were photographed with a Keeler-Konan contact specular microscope and a Konan Noncon Robo automated noncontact specular microscope. (i) After measuring and calibrating the magnification of each instrument, we digitized the cellular apices and analyzed the images from both instruments by using the Corners Method modified to accept x and y calibrations. (ii) Using the internal calibration marks of the Konan Noncon Robo specular microscope for calibration of magnification (as required for the Center Method), we evaluated identical cells on images from this microscope by both the Center Method and the Corners Method. (iii) We evaluated the reproducibility of both methods by repeating measurements on the same image. RESULTS: (i) When the images were properly calibrated for magnification by using an external scale, endothelial cell density (ECD) of normal corneas was 2,703 +/- 354 (mean +/- SD) cells/mm2 by contact and 2,685 +/- 357 cells/mm2 by noncontact techniques (p = 0.51). ECD of PK corneas was 1,767 +/- 773 cells/mm2 by contact and 1,807 +/- 775 cells/mm2 by noncontact techniques (p = 0.31). (ii) When images from the Konan Noncon Robo specular microscope were calibrated for magnification on the internal marks, the measured ECD from the same noncontact photographs was 6% less (p < 0.001). ECD was then 2,519 +/- 294 cells/mm2 (means +/- SD) by the Center Method and 2,523 +/- 305 cells/mm2 by the Corners Method (p = 0.55) in normal corneas and 1,715 +/- 748 cells/mm2 by the Center Method and 1,731 +/- 763 cells/mm2 by the Corners Method (p = 0.04) in PK corneas. (iii) The coefficient of variation of repeated measurements on the same normal image was 0.0025 for the Centers Method and 0.0099 for the Corners Method. CONCLUSIONS: (i) Images from the automated noncontact specular microscope may be used interchangeably with those from the contact specular microscope to measure ECD, but only if both are properly calibrated by measuring an external scale. (ii) As a method of analysis, the Center Method is equivalent to the Corners Method in normal corneas, but the proprietary internal calibration of the Center Method, which is required for its use, yields ECDs approximately 6% less than when an external scale is used for distance calibration. (iii) Cell density measurements by both the Center Method and the Corners Method were reproducible within 1%.

Cell Count↗
PubMed1999

Mutation causing congenital myasthenia reveals acetylcholine receptor beta/delta subunit interaction essential for assembly.

We describe a severe postsynaptic congenital myasthenic syndrome with marked endplate acetylcholine receptor (AChR) deficiency caused by 2 heteroallelic mutations in the beta subunit gene. One mutation causes skipping of exon 8, truncating the beta subunit before its M1 transmembrane domain, and abolishing surface expression of pentameric AChR. The other mutation, a 3-codon deletion (beta426delEQE) in the long cytoplasmic loop between the M3 and M4 domains, curtails but does not abolish expression. By coexpressing beta426delEQE with combinations of wild-type subunits in 293 HEK cells, we demonstrate that beta426delEQE impairs AChR assembly by disrupting a specific interaction between beta and delta subunits. Studies with related deletion and missense mutants indicate that secondary structure in this region of the beta subunit is crucial for interaction with the delta subunit. The findings imply that the mutated residues are positioned at the interface between beta and delta subunits and demonstrate contribution of this local region of the long cytoplasmic loop to AChR assembly.

Acetylcholinesterase↗
PubMed1999

Effects of NTE-122, a novel acyl-CoA:cholesterol acyltransferase inhibitor, on cholesterol esterification and secretions of apolipoprotein B-containing lipoprotein and bile acids in HepG2.

We studied the effect of NTE-122 (trans-1,4-bis[[1-cyclohexyl-3-(4-dimethylamino phenyl) ureido]methyl]cyclohexane), a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, on intracellular cholesterol esterification and the secretion of apolipoprotein B100 (apoB)-containing lipoprotein and bile acids in the human hepatoma cell line HepG2. NTE-122 markably inhibited [3H]oleate incorporation into cholesteryl esters in HepG2 cells incubated with 5 microg/ml 25-hydroxycholesterol as a stimulus for ACAT (IC50=6.0 nM). On the other hand, NTE-122 did not affect [3H]oleate incorporation into triglycerides and phospholipids and [14C]acetate incorporation into cholesterol. The stimulation of ACAT by 25-hydroxycholesterol caused significant increases in the secretion of radiolabeled cholesteryl esters, radiolabeled triglycerides and apoB mass. NTE-122 pronouncedly inhibited the secretion of radiolabeled cholesteryl esters in proportion to the inhibition of cellular cholesterol esterification, and it significantly reduced the secretion of radiolabeled triglycerides and apoB mass in HepG2 cells incubated with 25-hydroxycholesterol. Furthermore, NTE-122 increased the secretion of bile acids synthesized from [14C]-cholesterol. These results suggest that NTE-122 is capable of exhibiting anti-hyperlipidemic effects by reducing both the cholesterol content and the amount of secreted very low-density lipoprotein and enhancing the excretion of bile acid from the liver.

Acetates↗
PubMed1999

Effects of NTE-122, a novel acyl-CoA:cholesterol acyltransferase inhibitor, on cholesterol esterification and high-density lipoprotein-induced cholesterol efflux in macrophages.

We investigated the effects of a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, NTE-122 (trans-1,4-bis[[1-cyclohexyl-3-(4-dimethylamino phenyl)ureido]methyl]cyclohexane), on ACAT activities in macrophages originating from several species and high-density lipoprotein (HDL)-induced cholesterol efflux in phorbol 12-myristate 13-acetate (PMA)-treated THP-1 cells. NTE-122 inhibited cell-free ACAT activities in human PMA-treated THP-1 cells and mouse J774.1 cells with IC50 values of 0.88 and 360 nM, respectively. NTE-122 competively inhibited the ACAT activity in PMA-treated THP-1 cells. NTE-122 also inhibited cellular ACAT activities in PMA-treated THP-1 cells, rat peritoneal macrophages and J774.1 cells with IC50 values of 3.5, 84 and 6800 nM, respectively. Furthermore, NTE-122 prevented cholesterol accumulation in PMA-treated THP-1 cells incubated with acetylated low density lipoprotein, simultaneously with HDL, while it caused accumulation of a significant amount of free cholesterol in the absence and even in the presence of HDL. NTE-122 also enhanced HDL-induced cholesterol efflux from established foam cells converted from PMA-treated THP-1 cells. These results suggest that NTE-122, capable of inhibiting macrophage ACAT activity in humans more strongly than those in the other species, exhibits anti-atherogenic effects by preventing the foam cell formation and enhancing the foam cell regression in humans.

Aniline Compounds↗
PubMed1999

Effects of NTE-122, an acyl-CoA:cholesterol acyltransferase inhibitor, on cholesterol esterification and lipid secretion from CaCo-2 cells, and cholesterol absorption in rats.

The effect of NTE-122 (trans-1,4-bis[[1-cyclohexyl-3-(4-dimethylamino phenyl)ureido]methyl]cyclohexane), an acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, on cholesterol absorption was investigated. NTE-122 inhibited whole-cell ACAT activity in CaCo-2 cells, a human intestinal cell line, with an IC50 value of 4.7 nM. In CaCo-2 cells cultured on a membrane filter, NTE-122 pronouncedly inhibited the basolateral secretion of newly synthesized cholesteryl esters, and significantly reduced the basolateral secretion of newly synthesized triglycerides without influencing the cellular triglyceride synthesis. Furthermore, NTE-122 (1 mg/kg, p.o.) inhibited [14C]cholesterol absorption in rats. These results suggest that NTE-122 is capable of exhibiting anti-hyperlipidemic effects by reducing the absorption of dietary cholesterol.

Aniline Compounds↗
PubMed1999

Serosurvey of Ehrlichia canis and Hepatozoon canis infection in dogs in Yamaguchi Prefecture, Japan.

Antibodies to Ehrlichia canis and Hepatozoon canis in dogs at the Animal Hospital in Yamaguchi University were surveyed and potential risk factors for both pathogens were evaluated. Among 430 dogs examined, 20 (4.7%) and 18 (4.2%) dogs showed positive findings for E. canis and H. canis, respectively. Neither, sex nor age was associated with the seropositivity of either pathogen, but the positive rate in dogs kept outside was slightly higher than that in dogs kept inside for both pathogens. A higher seropositive reaction to E. canis and H. canis was observed in dogs that lived in certain cities and towns. Beagles, golden retrievers and pointers had higher seropositivity than other breeds in E. canis, whereas shibas, akitas, beagles, pointers and mongrels had higher positive rates than other breeds in H. canis.

Age Factors↗
PubMed1999

Cloning of feline cDNA encoding the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4).

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) is a CD28 homologue which down-modulate T cell responses rather than augment them. To investigate its biological role in feline immune system, we cloned and sequenced full-length feline CTLA-4 (fCTLA-4) cDNA by RT-PCR from pokeweed mitogen stimulated peripheral blood lymphocytes. The fCTLA-4 contains an open reading frame of 669 nucleotides, coding for a polypeptide of 223 amino acids. The predicted fCTLA-4 amino acids sequence shows the homology of 86.6%, 87.0%, and 76.2% with human, bovine, and murine molecules respectively. The hexapeptide motif (MYPPPY) within the extra-cellular domain of CTLA-4 molecule, which is believed to be responsible for interaction with the B7 family members, is completely conserved in all the species.

Abatacept↗
PubMed1999

Duration of antibodies against 24 kd protein of Rhipicephalus sanguineus extract in dogs infested with the adult ticks.

A 24 kd protein from Rhipicephalus sanguineus (Rs24p) which was common to larvae, nymphs, male and female whole body and salivary gland extract of males and female was detected specifically in the serum from dogs after repeated infestation with adult R. sanguineus. The duration of antibodies against Rs24p in dogs infested with adults was examined by Western blotting analysis. Anti-Rs24p antibody was detected in two of 4 dogs during the period of 40 days in the first infestation. In the second infestation, all dogs showed positive reaction against Rs24p, but the duration of the antibodies varied greatly among the animals.

Animals↗
PubMed1999

Expression of recombinant feline serum amyloid A (SAA) protein.

Feline serum amyloid A (SAA) cDNA clone was isolated from a hepatic mRNA of a mixed-breed cat. The feline SAA cDNA clone contains 333 nucleotides and deduced amino acid sequence shows 87.4%, 73.9%, and 65.8% homology with dog, human and mouse SAA respectively. Relative to the human and mouse SAA proteins, an additional peptide of eight amino acids is specified in the feline cDNA clone. Recombinant feline SAA (rfSAA) was expressed at high levels using pGEX bacterial expression system. Cleavage from the fusion moiety, and purification using glutathione-sepharose yielded pure soluble form of rfSAA. Antibodies generated against rfSAA were specific for feline SAA and showed no cross-reactivity with human SAA. Furthermore, antibodies against human SAA did not react with feline SAA. These results indicate that antigenicity of feline SAA is totally different from human SAA.

Amino Acid Sequence↗
PubMed1999

Cloning of a full length cDNA encoding canine Interleukin-4.

Primer designed on the basis of the conserved transcription start point (tsp) region of human and bovine gene transcripts, encoding the Interleukin-4 (IL-4), and subsequently the gene specific primer and an adaptor primer pair, was successfully used, to generate the full-length complementary DNA (cDNA) sequence coding for canine IL-4 (cIL-4), from pokeweed mitogen stimulated canine peripheral blood lymphocytes. The full-length cIL-4 contains an open reading frame of 399 nucleotides (nt), with a 5' ends of 66 base pairs (bp) and 3' ends of 125 bp. The nucleotide sequence contains six possible Asn-X-Thr or Asn-X-Ser linked glycosylation sites. Five sequence motifs of TATT or ATTTA, responsible for the regulation of gene expression, are found in the 3' untranslated region.

Amino Acid Sequence↗
PubMed1999

Local mitochondrial function following traumatic brain injury in rats.

The effect of lateral fluid percussion injury on mitochondrial function in the rat brain was investigated by quantitative imaging of changes in the regional activity of succinate dehydrogenase (SDH), a mitochondrial enzyme of the tricarboxylic acid cycle for adenosine triphosphate production. Regional SDH was measured in the frontal, parietal, temporal, and occipital cortices, CA1 and CA2-3 of the hippocampus, thalamus, corpus callosum, caudate/putamen, and cerebellum 1 hour and 72 hours after low, medium, and high pressure injury. No regional difference between the hemispheres in the activity of SDH was observed in the sham group. The hippocampus showed high SDH activity. The CA2-3 regions showed the highest activity among the regions examined. The corpus callosum, which is white matter, showed the lowest. One hour after low pressure fluid percussion injury, only the frontal lobe showed significantly lower SDH activity than the sham control in the ipsilateral hemisphere, whereas after 72 hours SDH activity was significantly lower in the frontal, parietal, and temporal lobes. SDH activity was significantly lower in the frontal, parietal, and temporal lobes in the medium and high pressure injury groups than in the sham control 1 hour after injury, and SDH activity in the CA1 and CA2-3 of the hippocampus was significantly decreased 72 hours after injury. No decrease in SDH activity was observed in any region of the contralateral hemisphere either 1 hour or 72 hours after injury. Mitochondrial dysfunction of the ipsilateral cortex and hippocampus following fluid percussion injury is correlated with the severity of injury and advances with time after injury. The results suggest that progression of mitochondrial dysfunction is associated with secondary bioenergetic deterioration.

Animals↗
PubMed1999

Successful transfer of localized autoimmunity with positively selected CD4+ cells to scid mice lacking functional B cells.

T-cell function of athymic BALB/c-nu/nu (nude) mice can be corrected by implantation of a embryonic rat thymus graft (TG) under the renal capsule (TG nude mice). However, multiple organ-localized autoimmune diseases, such as oophoritis, Sjögren's syndrome like disease and gastritis, develop spontaneously in TG nude mice. Transfer of spleen cells from TG nude mice with such diseases leads to multiple localized autoimmune lesions with appearance of the corresponding autoantibodies in the recipient C.B-17-scid (SCID) mice. In the present study, removal of CD90+ or CD4+, but not CD8+ cells eliminated the transfer activity. Positively selected CD4+ cells proved capable of inducing lesions without the appearance of organ-specific autoantibodies, although the grade of lesions was lower than that in recipient mice that received untreated or CD8-depleted spleen cells. Target organs demonstrating CD4+ cell infiltration, generally expressed major histocompatibility complex (MHC) class II antigen (Ia) on their parenchymal cells. Injection of sera from TG nude mice with autoantibodies to SCID mice did not induce any pathogenic features in the autoantibody-target organs, although deposition of immunoglobulins in the corresponding target organs was observed. In such cases, no Ia antigens were expressed on the parenchymal cells. The data thus indicate that effector CD4+ cells can induce autoimmunity without B-cell help but that cooperation with functional B cells induces more severe tissue damage.

Animals↗
PubMed1999

[Analysis of outcome of free jejunal-autograft for head and neck reconstruction--postoperative complications and functional results of swallowing in 49 cases].

We evaluated postoperative results in 49 patients (39 men, 10 women) who underwent pharyngoesophageal reconstruction with free-jejunal autograft following total pharyngolaryngoesophagectomy in the Department of Otolaryngology, Hokkaido University School of Medicine from 1989 to 1997. Evaluation was performed regarding the following points: 1) postoperative complications, 2) factors that determine the functional results of swallowing, 3) relation between forms of jejunal anastomosis and swallowing. The primary malignancy site was hypopharynx (39), cervical esophagus (4), larynx (3), thyroid (2) and trachea (1), Reconstructions were made with free jejunal autograft alone in 45 cases and with free jejunum in combination with gastric pull-up in 4. In patients who underwent reconstruction with jejunum alone, the anastomosis of the jejunum to the pharynx was performed in side-to-end fashion in 22, end-to-end in 18 and rho-shaped in 4. In the 1 remaining case, we used jejunal-patch graft. Postoperative complications including minor or nongraft related, occurred in 24 of 49 (49.0%) patients. Among these, graft-related complications were graft failure in 1 (2.0%), fistula formation in 3 (6.1%) and graft stricture in 2 (4.0%). Re-operations were required in one case of graft failure and 2 of fistula formation. Consequently, the overall graft-survival rate was 98.0% (48/49). Therefore, we considered the method of reconstruction to be a reliable procedure with a high-success rate. The swallowing function after reconstructive surgery was studied in 35 patients who underwent side-to-end (18) and end-to-end (17) anastomosis of the jejunum to the pharynx. We indicated that appropriate tension in the jejunum was the most important factor for adequate swallowing function. The end-to-end group had a higher rate of taking normal diet compared with the side-to-end group. The rate of swallowing dysfunction was only 5.9% (1/17) in the end-to-end group. On the other hand, 4 of 18 (22.2%) cases in the side-to-end group were regarded as having poor swallowing function. As a result we considered the end-to-end proximal jejunal anastomosis to be the more desirable form.

Adult↗
PubMed1999

[Combined multimodal therapy for osteosarcoma--neoadjuvant chemotherapy].

Until the 1970s, the survival rate of osteosarcoma patients was less than 20%. By the 1990s, this had improved to 60% to 70%, and limb-sparing procedures have replaced amputation in many patients thanks to effective combination therapy. Neoadjuvant chemotherapy has become an accepted practice in the majority of institutions using protocols which include MTX, ADR, BCD and CDDP as the most active agents against this disease. Newer agents, particularly IFM and ETP, are increasingly incorporated into complex regimens. While several studies have reported multivariate analyses to identify prognostic factors, the histologic response to preoperative chemotherapy remains the most important prognostic factor. Pulmonary metastases are the primary cause of death in patients with osteosarcoma. Although current treatment regimens allow effective salvage therapy for the patients with pulmonary metastases, the actuarial survival rate is 30%. A more effective systemic treatment for those patients is needed. The current management of osteosarcoma is critically reviewed and a treatment strategy is proposed for discussion.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1999

[Application of microvascular surgery in reconstructive digestive tract surgery].

In our institutes, microvascular surgery has been effectively used in reconstructive digestive tract surgery, including esophageal reconstruction and hepatic arterial reconstruction. Free jejunal transfer combined with a gastric pedicle or microvascularly augmented elongated gastric pedicle has been utilized for total esophageal reconstruction. A microvascularly augmented jejunal pedicle or colonic pedicle has been applied in thoracic esophageal reconstructive cases with gastrectomy. Moreover, microvascular surgery has been performed in the reconstruction of the hepatic arterial system in the surgical treatment of pancreatic or bile duct cancer and living related-donor liver transplantation. Some pitfalls in selection of the recipient vessels and handling the intraperitoneal vessels for microvascular anastomosis are also described. Although microvascular surgery has been carried out by plastic and reconstructive surgeons in a team surgical approach, revisions in the medical educational system to create a new-type of surgeon with practical skills and clinical experience in both digestive tract and microvascular surgery will be required in future.

Aged↗
PubMed1999

[Surgical analysis for small cell lung cancer of the lung].

Clinical features of small cell lung cancer were studied in 15 cases. The overall 5-year survival rate of the patients with limited small cell lung cancer was 11.4%. Surgery played substantial role for long-term survival in limited SCLC. The 4-year-survival rate of the patients in stage I was 50%, and that of those in stage II and IIIA was 50% and 37.5%, respectively. In the two survivors over four years in stage IIIA, all tumor was categorized as pT3 disease. The 4-year-survival rate of the patients treated with PE was 100%, and that of those treated with another chemotherapy was 10%, and that difference is statistically significant (p < 0.05). There was no significant difference in prognosis of patients in any other factors such as location (central or peripheral), histological subtype, curability or R number, pT factor, pN factor, p stage or with or without thoracic irradiation. Surgical resection for limited SCLC should be recommended in patients with stage I, II and T3N0M0 or T3N1M0 disease. For the patients in stage IIIA, particularly in N2M0 disease, who showed partial response or no change after chemotherapy, surgery should be considered because those patients might have nonsmall cell carcinoma components.

Adult↗
PubMed1999

Isolated histiocytosis X of the pituitary stalk.

A case of histiocytosis X granuloma localized in the pituitary stalk is reported. Coronal and sagittal magnetic resonance imaging views were useful to determine the precise size and location of the mass lesion. The diagnosis was established immunohistochemically and the patient was treated with low-dose irradiation therapy. After irradiation, the patient improved well without endocrine replacement treatment. The pituitary stalk recovered its normal size with no evidence of recurrence on MRI at 7-year follow-up. We emphasize the importance of MRI before initiating therapy to evaluate the pituitary mass lesion and the effectiveness of low-dose irradiation for isolated histiocytosis X.

Adult↗