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1.
Biochem Biophys Res Commun ; 589: 92-99, 2022 01 22.
Article in English | MEDLINE | ID: mdl-34896781

ABSTRACT

Heated tobacco products (HTPs) are an emerging class of tobacco goods that claim to have lower health risks than those of smoking combustible tobacco products. In this study, we exposed human lung epithelial cell lines to extracts prepared from HTP aerosols and combustible cigarette smoke to compare cytotoxicity. We focused on the effects of aldehydes present in the aerosols of HTPs at levels close to those in combustible cigarette smoke. Significant toxicity was confirmed for the HTP extract, albeit to a lesser extent than that with the combustible cigarette extract. When redox balance was evaluated by the oxidative loss of low-molecular-weight thiols in the cells, we found that total glutathione (GSH) contents and low-molecular-weight thiol levels were significantly decreased after exposure to the aerosol extract of HTPs. These results indicated that GSH is rapidly consumed during the detoxification of xenobiotics, such as aldehydes from tobacco extracts. Accordingly, exposure to the aerosol extract of HTPs resulted in the enhanced carbonylation of many proteins. In a simple comparison, the results for HTPs were significantly different from those obtained with combustible cigarette smoke, suggesting reduced toxicity of HTPs. However, we found significant and harmful effects after exposing lung epithelial cells to the aerosol extract of HTPs. Thus, a further comprehensive study is needed to clarify the lung damage induced via the long-term inhalation of aerosols from HTPs.


Subject(s)
Aerosols/adverse effects , Epithelial Cells/metabolism , Glutathione/metabolism , Hot Temperature , Lung/pathology , Nicotiana/chemistry , Protein Carbonylation , A549 Cells , Cell Death , Gases , Humans , Molecular Weight , Sulfhydryl Compounds/metabolism , Tobacco Products , Volatilization
2.
Ann Oncol ; 26(5): 966-973, 2015 May.
Article in English | MEDLINE | ID: mdl-25712457

ABSTRACT

BACKGROUND: Central nervous system (CNS) relapse is an uncommon but challenging complication in patients with mantle cell lymphoma (MCL). Survival after CNS relapse is extremely poor. Identification of high-risk populations is therefore critical in determining patients who might be candidates for a prophylactic approach. PATIENTS AND METHODS: A total of 608 patients (median age, 67 years; range 22-92) with MCL newly diagnosed between 1994 and 2012 were evaluated. Pretreatment characteristics and treatment regimens were evaluated for their association with CNS relapse by competing risk regression analysis. RESULTS: None of the patients received intrathecal prophylaxis. Overall, 33 patients (5.4%) experienced CNS relapse during a median follow-up of 42.7 months. Median time from diagnosis to CNS relapse was 20.3 months (range: 2.2-141.3 months). Three-year cumulative incidence of CNS relapse was 5.6% [95% confidence interval (95% CI) 3.7% to 8.0%]. Univariate analysis revealed several risk factors including blastoid variant, leukemic presentation, high-risk MCL International Prognostic Index and high Ki-67 (proliferation marker). Multivariate analyses revealed that Ki-67 ≥ 30 was the only significant risk factor for CNS relapse (hazard ratio: 6.0, 95% CI 1.9-19.4, P = 0.003). Two-year cumulative incidence of CNS relapse in patients with Ki-67 ≥ 30 was 25.4% (95% CI 13.5-39.1), while that in the patients with Ki-67 < 30 was 1.6% (95% CI 0.4-4.2). None of the treatment modalities, including rituximab, high-dose cytarabine, high-dose methotrexate or consolidative autologous stem-cell transplant, were associated with a lower incidence of CNS relapse. Survival after CNS relapse was poor, with median survival time of 8.3 months. There was no significant difference in the survival by the site of CNS involvement.


Subject(s)
Central Nervous System Neoplasms/chemistry , Ki-67 Antigen/analysis , Lymphoma, Mantle-Cell/chemistry , Adult , Aged , Aged, 80 and over , Central Nervous System Neoplasms/mortality , Central Nervous System Neoplasms/pathology , Central Nervous System Neoplasms/therapy , Female , Humans , Incidence , Japan/epidemiology , Kaplan-Meier Estimate , Lymphoma, Mantle-Cell/mortality , Lymphoma, Mantle-Cell/pathology , Lymphoma, Mantle-Cell/therapy , Male , Middle Aged , Multivariate Analysis , Predictive Value of Tests , Recurrence , Retrospective Studies , Risk Assessment , Risk Factors , Time Factors , Treatment Outcome , Young Adult
3.
Osteoarthritis Cartilage ; 21(9): 1179-84, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23973128

ABSTRACT

OBJECTIVES: Knee osteoarthritis (OA) pain is suggested to be associated with inflammation and detrimental mechanical loading across the joint. In this cross-sectional study, we simultaneously examined the inflammation and alignment of the lower limb and examined how the pain components varied depending on the disease progression. DESIGN: One-hundred sixty female medial type of early- [n = 74 in Kellgren-Lawrence (K/L) 2] to advanced-stage (n = 96 in K/L >2) knee OA subjects (70.5 years on average) were enrolled. Knee pain was evaluated using a pain visual analog scale (VAS) and the pain-related subcategory of the Japanese Knee Osteoarthritis Measure (JKOM-pain). The serum interleukin (sIL)-6 level reflecting synovitis, and the high sensitivity C-reactive protein (hs-CRP) level were measured to evaluate the severity of inflammation. The anatomical axis angle (AAA) was measured as an alignment index. The ß-coefficient was estimated after adjusting for age and the body mass index (BMI) using a multiple linear regression analysis. RESULTS: Multiple linear regression analyses showed that the sIL-6 levels, but not AAA, associated with the pain VAS [ß = 10.77 (95% confidence interval (CI): 4.14-17.40), P < 0.01] and JKOM-pain scores [ß = 3.19 (95% CI: 1.93-4.44), P < 0.001] in the early stage. Conversely, AAA, but not the sIL-6 levels, was found to be associated with the pain VAS [ß = -1.29 (95% CI: -2.51 to -0.08), P < 0.05] and JKOM-pain scores [ß = -0.49 (95% CI: -0.82 to -0.16), P < 0.01] in the advanced stage. CONCLUSIONS: The presence of a higher level of sIL-6 and the varus alignment of the joint is associated with pain in early- and advanced-stage knee OA patients, respectively.


Subject(s)
Arthralgia/diagnostic imaging , Arthralgia/epidemiology , Osteoarthritis, Knee/diagnostic imaging , Osteoarthritis, Knee/epidemiology , Severity of Illness Index , Aged , Arthralgia/physiopathology , Biomarkers/blood , C-Reactive Protein/metabolism , Cross-Sectional Studies , Disease Progression , Female , Humans , Interleukin-6/blood , Middle Aged , Osteoarthritis, Knee/physiopathology , Pain Measurement , Radiography , Risk Factors , Synovitis/diagnostic imaging , Synovitis/epidemiology , Synovitis/physiopathology , Weight-Bearing/physiology
4.
J Chem Phys ; 138(21): 214308, 2013 Jun 07.
Article in English | MEDLINE | ID: mdl-23758372

ABSTRACT

Electron reemission following photoelectron recapture due to post-collision interaction has been studied at 0.7 eV the inner-shell photoionization threshold of water molecules, using a multi-electron coincidence method. Electron reemissions after single Auger decay occur from O and OH fragments which are produced by the dissociations of high-n Rydberg H2O(+) states populated through photoelectron recapture. In addition, electron reemissions after double Auger decay are identified in triple coincidence events, where autoionization lines from O and O(+) fragments are observed.


Subject(s)
Electrons , Water/chemistry , Photochemical Processes
5.
East Mediterr Health J ; 19(11): 905-8, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24673079

ABSTRACT

Mental health services are far from satisfactory in the Eastern Mediterranean Region. The Global Mental Health Assessment Tool-Primary Care version (GMHAT/PC) is a semi-structured, computerized clinical assessment tool that was developed to assist health workers in making quick, convenient and comprehensive standardized mental health assessments. A study was carried out in the United Arab Emirates to evaluate the validity and feasibility of the Arabic version of the GMHAT/PC. Mental health nurses administered the GMHAT/PC Arabic version to 50 patients in mental health and rehabilitation settings and their GMHAT/PC diagnosis was compared with the psychiatrist's independent ICD-10 based clinical diagnosis on the same patients. The nurses found GMHAT/PC easy to administer in an average of 16 minutes. The GMHAT/PC-based diagnosis had a good agreement with the psychiatrist's diagnosis (kappa = 0.91) and a high sensitivity (97%) and specificity (94%).


Subject(s)
Diagnosis, Computer-Assisted/standards , Mental Disorders/diagnosis , Mental Health , Psychiatric Status Rating Scales/standards , Adolescent , Adult , Aged , Feasibility Studies , Female , Humans , Language , Male , Middle Aged , United Arab Emirates , Young Adult
6.
Ann Oncol ; 22(8): 1865-71, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21289367

ABSTRACT

BACKGROUND: Because of the less graft-facilitating effect by bone marrow (BM), we need to assess a dosage of conditioning more accurately particularly in combination with reduced-intensity conditioning. Thus we examined that modified continual reassessment method (mCRM) is applicable for deciding appropriate conditioning of allogeneic BM transplantation. PATIENTS AND METHODS: The conditioning regimen consisted of i.v. fludarabine (125 mg/m2) plus an examination dose of i.v. melphalan. The primary endpoint was a donor-type T-cell chimerism at day 28 with successful engraftment defined as >90% donor cells. Five patients per dose level were planned to be accrued and chimerism data were used to determine the next dose. RESULTS: Seventeen patients were enrolled at doses between 130 and 160 mg/m2. The dose was changed from 160 to 130 mg/m(2) (second level) after five full-donor chimerisms. With one patient of 0% chimera in the second level, the dose was increased to 135 mg/m2 (third level). Following five full-donor chimerisms in the third level, the study was complete as projected. CONCLUSIONS: mCRM was shown to be a relevant method for dose-finding of conditioning regimen. The melphalan dose of 135 mg/m2 was determined as the recommended phase II dose to induce initial full-donor chimerism.


Subject(s)
Bone Marrow Transplantation , Chimerism/drug effects , Hematologic Neoplasms/surgery , Melphalan/administration & dosage , Myeloablative Agonists/administration & dosage , Transplantation Conditioning/methods , Vidarabine/analogs & derivatives , Adult , Dose-Response Relationship, Drug , Female , Humans , Male , Middle Aged , T-Lymphocytes/drug effects , Tissue Donors , Transplantation, Homologous , Vidarabine/administration & dosage
7.
Bone Marrow Transplant ; 41(8): 729-36, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18176617

ABSTRACT

Chronic GVHD is a significant complication following allogeneic hematopoietic stem cell transplantation; however, the clinical characteristics of chronic GVHD following cord blood transplantation (CBT) in adults have not been well described. Between March 2001 and November 2005, a total of 77 patients underwent CBT at eight transplantation centers of the Nagoya Blood and Marrow Transplantation Group. Of 77 patients, 29 survived without graft failure or progression of underlying diseases for at least 100 days after transplantation. The median age of the 29 patients was 42 years (range, 18-67 years). Seven patients developed chronic GVHD (extensive, n=4; limited, n=3) disease. The cumulative incidence of chronic GVHD 1 year after day 100 was 24% (95% confidence interval (CI), 11-41%), and the organs involved were the skin (n=6), oral cavity (n=4), liver (n=1) and gastrointestinal tract (n=1). In three patients, chronic GVHD was resolved with supportive care. The remaining four were successfully treated with additional immunosuppressive therapy. Event-free survival rates of the 29 patients with and without chronic GVHD 3 years after day 100 were 83 (95% CI, 27-97%) and 36% (95% CI, 17-56%), respectively (P=0.047). These results suggest that chronic GVHD following CBT is mild and has a graft-versus-malignancy effect.


Subject(s)
Cord Blood Stem Cell Transplantation/adverse effects , Graft vs Host Disease/classification , Adolescent , Adult , Aged , Disease-Free Survival , Female , Graft vs Tumor Effect , Humans , Male , Middle Aged , Retrospective Studies , Severity of Illness Index
8.
Bone Joint J ; 100-B(5): 590-595, 2018 05 01.
Article in English | MEDLINE | ID: mdl-29701088

ABSTRACT

Aims: The aim of this study was to evaluate antegrade autologous bone grafting with the preservation of articular cartilage in the treatment of symptomatic osteochondral lesions of the talus with subchondral cysts. Patients and Methods: The study involved seven men and five women; their mean age was 35.9 years (14 to 70). All lesions included full-thickness articular cartilage extending through subchondral bone and were associated with subchondral cysts. Medial lesions were exposed through an oblique medial malleolar osteotomy, and one lateral lesion was exposed by expanding an anterolateral arthroscopic portal. After refreshing the subchondral cyst, it was grafted with autologous cancellous bone from the distal tibial metaphysis. The fragments of cartilage were fixed with 5-0 nylon sutures to the surrounding cartilage. Function was assessed at a mean follow-up of 25.3 months (15 to 50), using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot outcome score. The radiological outcome was assessed using MRI and CT scans. Results: The mean AOFAS score improved from 65.7 (47 to 81) preoperatively to 92 (90 to 100) at final follow-up, with 100% patient satisfaction. The radiolucent area of the cysts almost disappeared on plain radiographs in all patients immediately after surgery, and there were no recurrences at the most recent follow-up. The medial malleolar screws were removed in seven patients, although none had symptoms. At this time, further arthroscopy was undertaken, when it was found that the mean International Cartilage Repair Society (ICRS) arthroscopic score represented near-normal cartilage. Conclusion: Autologous bone grafting with fixation of chondral fragments preserves the original cartilage in the short term, and could be considered in the treatment for adult patients with symptomatic osteochondral defect and subchondral cysts. Cite this article: Bone Joint J 2018;100-B:590-5.


Subject(s)
Bone Cysts/surgery , Bone Transplantation , Cartilage Diseases/surgery , Cartilage, Articular/surgery , Talus/surgery , Adolescent , Adult , Aged , Bone Cysts/diagnostic imaging , Cartilage Diseases/diagnostic imaging , Cartilage, Articular/diagnostic imaging , Cartilage, Articular/injuries , Female , Humans , Male , Middle Aged , Talus/diagnostic imaging , Transplantation, Autologous , Young Adult
9.
Bone Marrow Transplant ; 39(1): 31-9, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17115066

ABSTRACT

Post transplant immune disorders are problematic in cord blood transplantation (CBT) for adult patients, and optimal prophylaxis has not been established. We investigated whether intensive graft-versus-host disease (GVHD) prophylaxis using short-term methotrexate (MTX) has a prognostic impact on CBT. Post-CBT immune reactions were classified according to time course as pre-engraftment immune reaction (PIR), engraftment syndrome (ES) or acute GVHD. Between March 2001 and November 2005, a total of 77 patients underwent CBT at eight transplantation centers. Median age was 48 years (range, 18-69 years). Preparative regimens comprised myeloablative (n=31) or reduced-intensity (n=46). Acute GVHD prophylaxis included cyclosporine alone (n=23), tacrolimus alone (n=12), cyclosporine plus MTX (n=17), tacrolimus plus short-term MTX (n=23) or cyclosporine plus methylprednisolone (n=2). Cumulative incidences of PIR, ES and grade II-IV GVHD were 36, 12 and 23%, respectively. Short-term MTX exerted significant favorable effects on post-CBT immune reactions (hazard ratio, 0.55; 95% confidence interval (95% CI), 0.31-0.98; P=0.04) in multivariate analysis. Overall survival rates for patients with and without short-term MTX at day 180 were 59% (95% CI, 42-73%) and 16% (95% CI, 6.6-30%) (P=0.0001), respectively. Short-term MTX could offer one optimal regimen to reduce immune reactions and improve outcomes in CBT.


Subject(s)
Cord Blood Stem Cell Transplantation , Graft vs Host Disease/mortality , Graft vs Host Disease/prevention & control , Immunosuppressive Agents/administration & dosage , Methotrexate/administration & dosage , Adolescent , Adult , Aged , Cord Blood Stem Cell Transplantation/mortality , Cyclosporine/administration & dosage , Disease-Free Survival , Female , Hematologic Neoplasms , Humans , Male , Middle Aged , Retrospective Studies , Survival Rate , Tacrolimus/administration & dosage , Time Factors , Transplantation, Homologous , Treatment Outcome
10.
Br J Ophthalmol ; 91(2): 161-5, 2007 Feb.
Article in English | MEDLINE | ID: mdl-16914470

ABSTRACT

AIM: To assess the efficacy and safety of an intravitreal injection of bevacizumab (Avastin(R)) for myopic choroidal neovascularisation (mCNV). METHODS: Intravitreal bevacizumab (1 mg) was injected into eight eyes of eight patients with mCNV in this non-randomised, interventional case series. The best-corrected visual acuity (BCVA) was measured and the optical coherence tomography (OCT) and fluorescein angiography findings were examined before and after treatment. The minimum follow-up time was 3 months. RESULTS: The mean BCVA was 0.26 before treatment and 0.51 at the last visit (p = 0.009). The BCVA improved to two or more lines in six eyes (75%) and remained the same in two eyes (25%). Leakage from the mCNV on fluorescein angiography decreased in seven eyes (87.5%). The choroidal neovascularisation area on fluorescein angiography (p = 0.049) and the foveal thickness on OCT images decreased significantly (p = 0.027) after the treatment. No major complications developed. CONCLUSION: Intravitreal injection of bevacizumab seems to be an effective and safe treatment for mCNV.


Subject(s)
Angiogenesis Inhibitors/therapeutic use , Antibodies, Monoclonal/therapeutic use , Choroidal Neovascularization/drug therapy , Myopia, Degenerative/complications , Adult , Aged , Angiogenesis Inhibitors/administration & dosage , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal, Humanized , Bevacizumab , Choroidal Neovascularization/etiology , Choroidal Neovascularization/pathology , Choroidal Neovascularization/physiopathology , Female , Follow-Up Studies , Fovea Centralis/pathology , Humans , Male , Middle Aged , Treatment Outcome , Vascular Endothelial Growth Factor A/antagonists & inhibitors , Visual Acuity/drug effects
11.
Bone Joint J ; 99-B(9): 1140-1146, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28860392

ABSTRACT

AIMS: Our aim was to evaluate the radiographic characteristics of patients undergoing total hip arthroplasty (THA) for the potential of posterior bony impingement using CT simulations. PATIENTS AND METHODS: Virtual CT data from 112 patients who underwent THA were analysed. There were 40 men and 72 women. Their mean age was 59.1 years (41 to 76). Associations between radiographic characteristics and posterior bony impingement and the range of external rotation of the hip were evaluated. In addition, we investigated the effects of pelvic tilt and the neck/shaft angle and femoral offset on posterior bony impingement. RESULTS: The range of external rotation and the ischiofemoral length were significantly lower, while femoral anteversion, the ischial ratio, and ischial angle were significantly higher in patients with posterior bony impingement compared with those who had implant impingement (p < 0.05). The range of external rotation positively correlated with ischiofemoral length (r = 0.49, p < 0.05), and negatively correlated with ischial length (r = -0.49, p < 0.05), ischial ratio (r =- 0.49, p < 0.05) and ischial angle (r = -0.55, p < 0.05). The range of external rotation was lower in patients with posterior pelvic tilt (p < 0.05) and in those with a high offset femoral component (p < 0.05) due to posterior bony impingement. CONCLUSION: Posterior bony impingement after THA is more likely in patients with a wider ischium and a narrow ischiofemoral space. A high femoral offset and posterior pelvic tilt are also risk factors for this type of impingement. Cite this article: Bone Joint J 2017;99-B:1140-6.


Subject(s)
Arthroplasty, Replacement, Hip , Femoracetabular Impingement/diagnostic imaging , Hip Prosthesis , Postoperative Complications/diagnostic imaging , Tomography, X-Ray Computed , Adult , Aged , Computer Simulation , Female , Femoracetabular Impingement/physiopathology , Humans , Imaging, Three-Dimensional , Male , Middle Aged , Postoperative Complications/physiopathology , Range of Motion, Articular
12.
Bone Marrow Transplant ; 52(6): 818-824, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28112749

ABSTRACT

Clinical efficacy of allogeneic hematopoietic cell transplantation (HCT) using reduced-intensity conditioning (RIC) for younger patients remains unclear. We therefore performed a retrospective registry-based study to evaluate outcomes for patients with AML aged between 16 and 49 years who underwent RIC allogeneic HCT. Patients receiving RIC (N=125) showed significantly worse survival than those receiving myeloablative conditioning (MAC; N=1,554) (47.7% for RIC and 54.2% for MAC at 4 years, P=0.047). However, the difference became marginal after adjustment for patient characteristics (P=0.080), and inclusion in the multivariate analysis of the HCT comorbidity index or the propensity score for estimating the likelihood of choosing RIC or MAC further reduced statistical significance (P=0.371 and 0.206, respectively), indicating the existence of a selection bias against RIC. Nevertheless, outcomes for our patients receiving RIC were still acceptable, so that RIC constitutes a potential therapeutic option for younger AML patients who are deemed unsuitable for MAC. Subgroup analyses showed that patients aged between 40 and 49 years as well as those in first or second CR at the time of transplantation exhibited similar outcomes regardless of whether they were treated with RIC or MAC.


Subject(s)
Hematopoietic Stem Cell Transplantation , Leukemia, Myeloid, Acute/mortality , Leukemia, Myeloid, Acute/therapy , Registries , Transplantation Conditioning , Adolescent , Adult , Allografts , Disease-Free Survival , Female , Humans , Male , Middle Aged , Survival Rate
13.
Vet J ; 214: 21-3, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27387721

ABSTRACT

Reproductive management is necessary to prevent deleterious genetic disorders in purebred dogs, but comprehensive studies aimed at prevention of multiple underlying genetic disorders in a single breed have not been performed. The aims of this study were to examine mutant allele frequencies associated with multiple genetic disorders, using Border collies as a representative breed, and to make recommendations for prevention of the disorders. Genotyping of known mutations associated with seven recessive genetic disorders was performed using PCR assays. More than half (56%) of the Border collies had no mutant alleles associated with any of the seven disorders, suggesting that these disorders can be removed from the population over several generations. Since frequencies of each mutant allele differed among disorders, reproductive management should be performed after the establishment of prevention schemes that are appropriate for each disorder, the type and specificity of genetic test available, and the effective population size in each breeding colony.


Subject(s)
Dog Diseases/epidemiology , Gene Frequency , Genetic Diseases, Inborn/veterinary , Animals , Breeding , Dog Diseases/genetics , Dogs , Genetic Counseling , Genetic Diseases, Inborn/epidemiology , Genetic Diseases, Inborn/genetics , Genotype , Japan/epidemiology , Mutation , Polymerase Chain Reaction/veterinary , Prevalence
14.
Eur J Ophthalmol ; 15(5): 641-2, 2005.
Article in English | MEDLINE | ID: mdl-16167296

ABSTRACT

PURPOSE: To report a peculiar curvilinear pigmentary lesion in the peripheral fundus in a rod-cone dystrophy. METHODS: Observational case report. Fundus examination of a 57-year-old woman who was known to have a generalized rod-cone dystrophy since she was 8 years old. RESULTS: The peripheral fundus examination revealed a curvilinear lesion which resembles a well-known finding associated with a presumed ocular histoplasmosis syndrome or multifocal choroiditis. CONCLUSIONS: The differential diagnosis of a peculiar curvilinear pigmentary lesion in the peripheral fundus may be expanded to include a generalized rod-cone dystrophy.


Subject(s)
Photoreceptor Cells, Vertebrate/pathology , Pigment Epithelium of Eye/pathology , Retinitis Pigmentosa/diagnosis , Choroiditis/diagnosis , Diagnosis, Differential , Eye Infections, Fungal/diagnosis , Female , Histoplasmosis/diagnosis , Humans , Middle Aged
15.
Anat Histol Embryol ; 44(4): 317-20, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25181932

ABSTRACT

Plastic hematocrit tubes (PHTs) are convenient tools for electron microscopy (EM) of peripheral blood buffy coats, and the PHT-EM technique is expected to be a practical method for veterinary clinical medicine. In this study, fixatives composed of various concentrations of sucrose, glutaraldehyde, and phosphate buffer (PB) were tested for preparing canine and feline buffy coats. The highest quality images were obtained using a fixative consisting of 2.5% glutaraldehyde in 0.1 m PB, and it was concluded that this method allows clinicians who are inexperienced in histological techniques can conveniently transport buffy coat samples to diagnostic laboratories for analysis by EM.


Subject(s)
Blood Buffy Coat/ultrastructure , Cats/blood , Dogs/blood , Microscopy, Electron/veterinary , Animals , Fixatives , Glutaral , Hematocrit/instrumentation , Hematocrit/veterinary , Microscopy, Electron/instrumentation , Microscopy, Electron/methods
16.
Transplantation ; 63(1): 131-5, 1997 Jan 15.
Article in English | MEDLINE | ID: mdl-9000674

ABSTRACT

This study aimed at investigating the efficacy of hepatic stimulatory substance (HSS) on the proliferation of transplanted hepatocytes in the spleen of rats with carbon tetrachloride (CCl4)-induced liver cirrhosis. After hepatocyte transplantation (HTx; 1x10(7) cells/rat), the recipients received intravenous administration of HSS (3ml) of 2, 4, or 7 times/week to investigate the effect of the frequency of HSS treatment on the proliferation of intrasplenically transplanted hepatocytes. Next, to investigate the effect of the severity of liver cirrhosis on HSS-stimulated proliferation of transplanted hepatocytes, different doses (0.00, 0.08, 0.12, and 0.16 ml/week) of CCl4 were given to rats after HTx. The recipients were killed and the spleens were removed at 2 and 4 weeks after HTx and stained with bromodeoxyuridine (BrdU) and hematoxylin and eosin for determining the BrdU labeling index (L.I.) and the hepatocyte-occupied area ratio in the longitudinal cut surface of the spleen (H:S ratio), respectively. The H:S ratio was measured in the hematoxylin and eosin-stained splenic sections under a light microscope connected to an image processor. HSS-treated rats showed significantly higher BrdU L.I. and H:S ratios than the untreated control rats at 2 and 4 weeks after HTx. However, the difference in the BrdU L.I. and H:S ratio was not statistically significant among the HSS-treated rats. Without HSS treatment, the severity of liver cirrhosis did not affect the proliferation of intrasplenically transplanted hepatocytes. On the other hand, HSS-induced proliferation of transplanted hepatocytes was further enhanced in proportion to the severity of liver cirrhosis. The H:S ratio of the rats treated with 0.04, 0.08, and 0.16 ml/week of CCl4 after HTx was 1.27+/-0.5%, 4.32+/-0.65%, and 6.25+/-0.70%, respectively, at week 4. During the long-term observation of up to 12 weeks, a marked decrease in the H:S ratio was observed in HSS-untreated control rats at weeks 4-12 compared with week 2. While HSS-treated rats revealed gradual proliferation of intrasplenically transplanted hepatocytes, the difference in the H:S ratio between HSS-treated and -untreated rats became larger. These results indicate that HSS treatment of recipient rats stimulated the proliferation of intrasplenically transplanted hepatocytes in cirrhotic rats and that the severity of liver cirrhosis enhanced the stimulative effect of HSS on the proliferation of transplanted hepatocytes.


Subject(s)
Growth Substances/pharmacology , Liver Cirrhosis, Experimental/surgery , Liver/cytology , Mitogens/pharmacology , Peptides/pharmacology , Animals , Bromodeoxyuridine/metabolism , Carbon Tetrachloride/toxicity , Cell Division/drug effects , Cell Transplantation , Intercellular Signaling Peptides and Proteins , Liver/drug effects , Male , Rats , Rats, Wistar , Spleen/cytology , Swine , Transplantation, Heterotopic
17.
Transplantation ; 62(2): 167-73, 1996 Jul 27.
Article in English | MEDLINE | ID: mdl-8755811

ABSTRACT

Dibutyryl cAMP (DBcAMP) has a high membrane permeability, and maintenance of the intracellular cAMP concentration may improve the viability of organs. In this study, the effect of DBcAMP pretreatment on warm ischemic injury of rat livers was evaluated. Warm ischemic liver injury was induced in adult Wistar rats weighing 250-280 g by leaving them at room temperature (22-25 degrees C) after cardiac arrest. The hepatic cAMP concentration, %ATP, and trypan blue-positive nuclear ratio were determined after different durations of warm ischemia. In addition, transaminase and endothelin-1 (ET-1) release into the perfusate were examined during 60 min of isolated liver perfusion with Krebs-Henseleit solution. The optimal dose and time of DBcAMP pretreatment were determined to be 15 mg/kg and 60 min prior to warm ischemia, respectively. Data on the trypan blue-positive nuclear ratio and the release of transaminases and ET-1 revealed that warm ischemia first damaged the endothelial cells and then the hepatocytes. DBcAMP pretreatment appeared to protect the liver from warm ischemic injury by increasing the intracellular cAMP concentration and stabilizing the cell membranes of endothelial cells and hepatocytes.


Subject(s)
Bucladesine/therapeutic use , Heart Arrest/physiopathology , Ischemia/prevention & control , Liver/blood supply , Adenosine Triphosphate/metabolism , Animals , Cyclic AMP/metabolism , Dose-Response Relationship, Drug , Endothelins/metabolism , Liver/metabolism , Liver/pathology , Male , Rats , Rats, Wistar , Temperature , Time Factors
18.
Invest Ophthalmol Vis Sci ; 28(9): 1457-63, 1987 Sep.
Article in English | MEDLINE | ID: mdl-3623832

ABSTRACT

The corneal endothelial permeability coefficient (Pac) for fluorescein and fluorescein glucuronide was determined in ten normal young volunteers. After oral administration of fluorescein, the apparent concentrations of both dyes in the corneal stroma and the anterior chamber were measured by differential fluorometry. The apparent dye levels calculated directly from the in vivo fluorometric measurements were converted to the true ones, based on the result of a normalization experiment performed in rabbit eyes. The value of Pac averaged 5.44 +/- 1.77 X 10(-4) cm/min for fluorescein and 3.77 +/- 1.10 X 10(-4) cm/min for fluorescein glucuronide (mean +/- SD, N = 20); the former was significantly greater than the latter (paired t-test, P less than 0.001). The aqueous-cornea distribution ratio was 0.50 +/- 0.14 for fluorescein and 0.66 +/- 0.16 for fluorescein glucuronide; the latter was significantly greater than the former (paired t-test, P less than 0.001). It was suggested that the previously reported values of Pac for fluorescein in the human eye were underestimates.


Subject(s)
Cornea/metabolism , Fluoresceins/metabolism , Administration, Oral , Adult , Anterior Chamber/metabolism , Endothelium/metabolism , Fluorescein , Fluorometry , Humans , Osmolar Concentration , Permeability
19.
Invest Ophthalmol Vis Sci ; 26(4): 443-9, 1985 Apr.
Article in English | MEDLINE | ID: mdl-3980166

ABSTRACT

After a mechanical denudation of rabbit corneal endothelial cells, the healing process was followed with wide-field specular microscopy. Individual cell migration and morphologic changes were analyzed by computer-assisted morphometry. The cells surrounding the wound migrated to cover the defect without producing intercellular gaps. The greatest cellular migration and morphologic alterations occurred close to the wound edge. As the cells migrated toward the wound, they elongated and increased their surface area in the direction of the migration. As the healing proceeded, the cells lost their original hexagonal pattern, which returned after coverage was complete. The wound was covered completely by large, irregularly shaped cells showing mitotic figures between 24 and 48 hr. During this period, cellular migration decreased and normal cellular morphology began to recover. When mitosis decreased, the normal cellular pattern rearranged towards a more hexagonal shape. During the healing process, the degree and direction of cellular migration varied from cell to cell. Additionally, changes in cell-to-cell contact (positional changes of neighboring cells) occurred in one-third of migrating cells. Such cellular migration can account for monolayered cells sliding without producing gaps between individual cells.


Subject(s)
Cornea/cytology , Wound Healing , Animals , Cell Communication , Cell Count , Cell Movement , Corneal Injuries , Endothelium/cytology , Rabbits , Time Factors
20.
Cell Transplant ; 5(5 Suppl 1): S35-7, 1996.
Article in English | MEDLINE | ID: mdl-8889227

ABSTRACT

We have investigated the availability of multiporous microcarriers (MCs) for immobilizing isolated rat hepatocytes, but the pore size of MCs was too small (35 microns) for hepatocyte immobilization. In this study, we immobilized isolated rat hepatocytes on MCs with larger pores, and evaluated their metabolic activity. Isolated hepatocytes were immobilized on MCs precoated with collagen by the intermittent stirring method and by aspiration, and the cell-protein content per 100 mg MCs was determined for comparison of these methods. Metabolic activity was evaluated by analyzing NH3 metabolism, urea nitrogen synthesis and glucose synthesis. The aspiration method immobilized significantly more of hepatocytes on MCs than the intermittent stirring method (p < 0.05). A stationary culture of hepatocytes immobilized on MCs showed a similar NH3 metabolism to monolayer cultured hepatocytes, and hepatocytes immobilized on MCs in a floating culture showed significantly higher NH3 metabolism than those in a stationary culture (p < 0.01). However, monolayer cultured hepatocytes showed higher glucose synthesis than hepatocytes immobilized on MCs in a stationary culture (p < 0.01). In conclusion, hepatocytes immobilized on MCs proved to be useful as a bioreactor in a hybrid artificial liver.


Subject(s)
Capsules , Liver/cytology , Liver/metabolism , Ammonia/metabolism , Animals , Cell Transplantation/instrumentation , Cell Transplantation/methods , Cells, Immobilized , Liver, Artificial , Male , Proteins/metabolism , Rats , Rats, Wistar
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