LAMB3 gene transfection into SV40-transformed keratinocytes from patient with Herlitz junctional epidermolysis bullosa.
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Cell Line, Transformed↗
Biomedical subjects
H Shimizu
Publications and source records attributed to H Shimizu.
The role of beta-tricalcium phosphate in vascularized periosteum.
To investigate the osteogenic capacity of vascularized periosteum in bore grafting, we prepared experimental groups by wrapping beta-tricalcium phosphate (beta-TCP) with vascularized periosteum of the femur of 12-week-old Japanese white rabbits, and evaluated osteogenesis histologically and biochemically. Bone formation was observed in the group with vascularized periosteum and in the group with bone marrow fluid added to the vascularized periosteum. In particular, woven bone was observed in the group that had added bone marrow. Osteogenesis appeared earlier in the group with bone marrow fluid added to the vascularized periosteum, but histologically, there was no significant difference between this group and the group with vascularized periosteum without bone marrow fluid added at week 24 after the operation. In the group with non-vascularized periosteum, slight osteogenesis was found at week 6 after the operation, but in the control group, with beta-TCP implanted in soft tissue, osteogenesis did not occur at all. Alkaline phosphatase (ALP) activity reached a peak at week 2 after the operation in the group with vascularized periosteum, but only half the peak value was then maintained until week 8. In the group with bone marrow fluid added to the vascularized periosteum, similar values were found from immediately after the operation until week 8. ALP activity did not show any significant difference between these two groups at week 8 postoperatively. In the group with beta-TCP implanted in the soft tissue, ALP activity was low at all times measured. These results suggested that the periosteum had osteoinduction capacity and beta-TCP had osteoconduction capacity; that better osteogenesis occurred with vascularized periosteum; and that bone marrow fluid was involved in the promotion of osteoblastic activity, but not in calcification.
Alkaline Phosphatase↗
Complications associated with clamping the aorta between the left common carotid artery and left subclavian artery.
BACKGROUND: Hypothermic circulatory arrest using a left thoracotomy has recently been recommended for repair of distal arch lesions to prevent the atheroembolism that often results from clamp injury. The recommendation holds even for cases in which aortic cross-clamping between the left common carotid artery and left subclavian artery is possible. METHODS: Over the last 16 years, 69 patients underwent repair of the distal arch or descending thoracic aorta using distal perfusion with the proximal aortic clamp placed between the left common carotid and left subclavian artery. The average age of the patients was 61+/-12 years; 18 of them (26%) were older than 70 years. Forty-four patients (64%) had atherosclerotic true aneurysms. RESULTS: The surgical procedures used included patch closure of saccular aneurysms in 20 patients (29%) and graft replacement in 47 (71%). The left subclavian artery was reattached in 7 patients (10%). Although there were 3 hospital deaths (4%), no cerebral complications occurred aside from temporary neurologic dysfunction in 4 patients (6%). CONCLUSIONS: An acceptably low incidence of cerebral complications is associated with cross-clamping the aorta between the left common carotid artery and left subclavian artery.
Adult↗
Selective perfusion of segmental arteries in patients undergoing thoracoabdominal aortic surgery.
BACKGROUND: Reattachment of segmental arteries is one method used to prevent paraplegia associated with thoracoabdominal aortic repair. Nevertheless, even when important segmental arteries are reattached, ischemia causing spinal injury may occur during anastomosis. METHODS: In 27 patients undergoing thoracoabdominal aortic repair, we attempted to perfuse the segmental arteries to be reattached with catheters connected to the distal bypass circuit. To identify perioperative risk factors for spinal ischemia, we examined changes in spinal somatosensory evoked potentials. RESULTS: A median value of four segmental arteries were perfused in 20 (74%) of the 27 patients. Changes in somatosensory evoked potential indicative of spinal ischemia were observed in 13 patients (48%). The only risk factor associated with changes in evoked potentials revealed by a multivariate analysis was prolonged aortic cross-clamp time (> 120 minutes). Of the 2 patients who suffered paraplegia, one had the longest clamp time and the other showed spinal cord necrosis due to embolic shower. CONCLUSIONS: Despite selective perfusion of segmental arteries, spinal ischemia associated with aortic cross-clamping may occur when clamping is prolonged over 120 minutes. Most of the changes appear to be reversible, however.
Adult↗
Cerebral complications associated with selective perfusion of the arch vessels.
BACKGROUND: Few studies have determined risk factors for postoperative cerebral complications associated with surgery of the aortic arch using selective cerebral perfusion. METHODS: Between November 1992 and December 1998, 113 patients underwent aortic arch repair combined with selective cerebral perfusion. For each patient, three arch vessels were perfused using a single roller pump at a rectal temperature of 23 degrees C. RESULTS: Among the 108 patients who underwent postoperative neurologic assessment, 25 patients (23%) suffered from cerebral complications. Five patients (5%) suffered from transient neurologic disturbance and 17 patients (16%) suffered from stroke, and 7 patients (7%) of the preceding 17 patients had residual neurologic disturbance upon discharge. Three patients (3%) with either preoperative coma (n = 1) or post bypass cardiac arrest (n = 2) sustained severe global cerebral dysfunction. The occurrence of cerebral complications was not related to cerebral perfusion time. Independent risk factors for cerebral complications included a history of cerebrovascular disease, perioperative shock, distal anastomosis below the left pulmonary artery, malperfusion of extremities, and older age (> 60 years). CONCLUSIONS: Although high-level brain function was well preserved in most patients, the incidence of stroke when using current selective cerebral perfusion techniques is still high.
Adult↗
Exogenous aspartate neurotoxicity in the spinal cord under metabolic stress in vivo.
BACKGROUND: Considerable evidence exists that neurotoxicity of excitatory amino acids is related to the neuronal injury, including paraplegia. However, little is known about aspartate neurotoxicity in the spinal cord in vivo. We evaluated the detrimental effects of exogenous aspartate on spinal cord neurons under metabolic stress. METHODS: New Zealand white rabbits underwent an infrarenal aortic isolation. Group A animals (n = 7) received segmental aspartate 50 mmol/L) infusion for 10 minutes. Group B animals (n = 7) received saline as a negative control. Group C animals (n = 5) received segmental aspartate 100 mmol/L) infusion for 5 minutes. Group D animals (n = 7) were pretreated with segmental infusion of (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cycloheptan-5,10-imine (MK-801) (6 mg/kg), a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist for 1 minute, followed by segmental infusion of aspartate (50 mmol/L) for 9 minutes. Group E animals (n = 7) received vehicle only, followed by aspartate (50 mmol/L) infusion as a control of group D. Neurologic status was assessed at 12, 24, and 48 hours after operation using the Tarlov score. RESULTS: Group A animals exhibited paraplegia or paraparesis with marked neuronal necrosis. Group B and C animals recovered fully. Group D animals showed significantly better neurologic function (p = 0.0007) compared with group E animals that exhibited paraplegia or paraparesis. CONCLUSIONS: Exogenous aspartate can have detrimental effects on spinal cord neurons under metabolic stress. This model may be useful in assaying neuronal injury mediated by NMDA receptor in vivo.
Animals↗
The role of protein-tyrosine phosphorylation and gelatinase production in the migration and proliferation of smooth muscle cells.
BACKGROUND: It has been reported that matrix metalloproteinase (MMP) was expressed in coronary arterial atherosclerotic lesions. However, not much is known about the relationship between the production of MMP and the progression of atherosclerosis. PURPOSE AND METHOD: To demonstrate the association between the protein-tyrosine phosphorylation (PTP) and the activation of extracellular MMP in the proliferation and migration of vascular smooth muscle cells (VSMCs), the effect of platelet-derived growth factor (PDGF) and vanadate (an inhibitor of protein-tyrosine phosphatase and an activator of certain protein-tyrosine kinases) on mitogenesis ([3H]thymidine incorporation after 24 hours), migration, PTP (Western blot analysis using anti-phosphotyrosine antibodies), and production of MMP (gelatin zymography) was examined in cultured VSMCs. RESULTS: Both vanadate (1-5 micromol/l) and PDGF (1-10 ng/ml) caused a dose-dependent increase in thymidine incorporation and migration and produced 72-kDa type IV gelatinase (MMP-2) in VSMCs. The combination of vanadate and PDGF resulted in a dose-dependent synergistic effect on thymidine incorporation and MMP-2 production. Western blot analysis revealed that PDGF caused an increase in PTP, extracellular signal-regulated kinases (ERK1, ERK2) and PDGF receptor in VSMCs. Vanadate given together with PDGF induced a marked increase in the intensity of tyrosine phosphorylation in these proteins. Tyrosine kinase inhibitors (genistein and herbimycin A) and a synthetic inhibitor of MMP (1,10-phenanthroline) and an anti-MMP-2 neutralizing antibody inhibited the mitogenic effect induced by vanadate and/or PDGF. CONCLUSIONS: The data suggest that the proliferation and migration of cultured VSMCs was closely related to the stimulation of MMP-2 production that was induced through activation of PTK.
Analysis of Variance↗
Cardiac rupture and admission electrocardiography in acute anterior myocardial infarction: implication of ST elevation in aVL.
This study determines the usefulness of electrocardiography in the emergency room for assessing the risk of cardiac rupture after acute anterior myocardial infarction (MI). The presence of ST segment elevation on the admission 12-lead electrocardiography was evaluated in 325 consecutive anterior MI patients. A forward-stepwise logistic regression analysis for cardiac rupture was performed with the covariates of age, gender, hypertension, history of MI, reperfusion therapy by coronary angioplasty, and ST segment elevations in leads I, aVL, V1-V6. Cardiac rupture occurred in 16 patients, including 7 with left ventricular free wall rupture (FWR) and 9 with ventricular septal perforation (VSP). For FWR, ST elevation in lead aVL was the only independent predictor (odds ratio = 12.1, P = .0215). For VSP, female gender (odds ratio = 5.32, P = .0201) was the independent predictor. In conclusion, in patients with acute anterior MI, ST segment elevation in lead aVL on the admission electrocardiography is a significant risk factor for left ventricular FWR.
Adult↗
Rationality/antiemotionality personality and selected chronic diseases in a community population in Japan.
OBJECTIVE: To clarify the relationships of the rationality/antiemotionality (R/A) personality with past histories of seven chronic diseases and current smoking habits in a community population in Japan. METHODS: A questionnaire survey was conducted of all residents aged 35 years old or over (n = 37,287) in Takayama city, Gifu prefecture, Japan; the response rate was 90.3%. Data from 13,091 males and 14,061 females who completed all relevant questions were analyzed. The subjects were asked to complete an 11-item R/A personality scale developed by Grossarth-Maticek et al. (1985). Past histories of seven major chronic diseases and current smoking habit were also asked in the questionnaire. RESULTS: Among males, the R/A scale score was significantly lower in those who had stroke, diabetes or allergy than in those who did not. Among females, the R/A scale score was significantly lower in those who had stroke, allergy or stomach cancer than in those who did not. Among females, current smokers had significantly lower R/A scores than non-smokers. After controlling for age, smoking and drinking, the R/A score was significantly and negatively associated with stroke among males and females; it was significantly and negatively associated with diabetes and allergy among males. CONCLUSION: Our findings are contrary to those observed in Yugoslavia and West Germany, and suggest a cultural difference in the relationship between the R/A personality and disease occurrence.
Adult↗
Mutation effects of a conserved threonine (Thr243) of cytochrome P450nor on its structure and function.
Threonine 243 of cytochrome P450nor (fungal nitric oxide reductase) corresponds to the 'conserved' Thr in the long I helix of monooxygenase cytochrome P450s. In P450nor, the replacement of Thr243 with Asn, Ala or Val makes the enzymatic activity dramatically reduce. In order to understand the roles of Thr243 in the reduction reaction of NO by P450nor, the crystal structures of three Thr243 mutants (Thr243-->Asn, Thr243-->Val, Thr243-->Ala) of P450nor were determined at a 1.4-A resolution and at cryogenic temperature. However, the hydrogen-bonding pattern in the heme pocket of these mutants is essentially similar for that of the WT enzyme. This suggests that the determination of the structure of the NADH complex of P450nor is required, in order to evaluate the role of Thr243 in its enzymatic reaction. We attempted to crystallize the NADH complex under several conditions, but have not yet been successful.
Amino Acid Substitution↗
Obstructive jaundice impairs hepatic sinusoidal endothelial cell function and renders liver susceptible to hepatic ischemia/reperfusion.
BACKGROUND/AIMS: Obstructive jaundice is associated with increased surgical morbidity and mortality. While parenchymal injury has been defined in obstructive jaundice, the pathogenesis of hepatic sinusoidal endothelial cell injury in obstructive jaundice is unclear. The aims of this study were to investigate hepatic sinusoidal endothelial cell injury in obstructive jaundice by determining serum hyaluronic acid levels, purine nucleoside phosphorylase/alanine aminotransferase ratios, and hyaluronic acid elimination rate, and also to determine whether hepatic parenchymal cell injury in obstructive jaundice is induced more than in normal liver after hepatic ischemia/reperfusion. METHODS: Male Wistar rats underwent ligation and division of the common bile duct (obstructive jaundice group) or sham operation (Sham group). Serum hyaluronic acid levels and purine nucleoside phosphorylase/alanine aminotransferase ratios in both groups were examined at intervals up to 21 days after surgery. Hepatic blood flow, permeability, neutrophil accumulation, and hyaluronic acid elimination rates in both groups were measured 14 days after surgery. Changes in serum hyaluronic acid and alanine aminotransferase concentrations were determined after 15 min of hepatic ischemia followed by reperfusion. RESULTS: Serum hyaluronic acid levels remained elevated after bile duct ligation. Hepatic sinusoidal endothelial cell swelling was observed by electron microscopy, and hepatic permeability was increased 14 days after bile duct ligation in association with neutrophil accumulation. Hepatic blood flow in obstructive jaundice remained unchanged, but hyaluronic acid elimination capacity was less than that in the Sham group. After hepatic reperfusion, the disappearance rate of serum hyaluronic acid in obstructive jaundice was lower, and serum alanine aminotransferase levels were higher than those in the Sham group. CONCLUSIONS: Our findings suggest that obstructive jaundice impairs sinusoidal endothelial cells and that sinusoidal endothelial cell damage in association with sinusoidal deterioration during obstructive jaundice renders liver susceptible to ischemia/reperfusion relative to normal liver.
Animals↗
Development of pemphigus vulgaris in a patient with pemphigus foliaceus: antidesmoglein antibody profile shift confirmed by enzyme-linked immunosorbent assay.
We describe a patient with pemphigus foliaceus (PF) in whom pemphigus vulgaris (PV) subsequently developed. The clinical change was accompanied by a shift of autoantibody profile confirmed by enzyme-linked immunosorbent assay. Antidesmoglein (Dsg) 1 antibodies alone were detected in the PF stage, whereas both anti-Dsg3 and anti-Dsg1 antibodies were detected in the PV stage.
Adult↗
Cultured keratinocytes from plectin/HD1-deficient epidermolysis bullosa simplex showed altered ability of adhesion to the matrix.
Epidermolysis bullosa simplex associated with late onset of muscular dystrophy has been found to show defective expression of plectin, an intracytoplasmic protein in hemidesmosomes. In this report, we examined ability of cell-to-matrix attachment of cultured keratinocytes derived from a case with this disease by various cell biological methods, and compared it to that of normal keratinocytes. In cell adhesion assay, the patient keratinocytes showed more prominent short-time cell adhesion than normal keratinocytes. In contrast, the patient keratinocytes could be detached much easier than normal keratinocytes in cell detachment assay by treatment with dispase. In phagokinetic track assay, no apparent difference of cell migration was observed between the patient and normal keratinocytes. These results indicate that plectin-deficiency may up-regulate short-term cell contact and reduce stable cell-matrix adhesion at the epidermal basement membrane zone.
Adult↗
Maximizing yellow pigment production in fed-batch culture of Monascus sp.
Yellow pigment production in exponential fed-batch cultivation of Monascus sp. was studied. Due to the difficulty of measuring the optical density for accurate determination of the cell concentration, a capacitance probe was employed on-line. The feed rate needed to keep the specific growth rate, mu, constant in fed-batch culture was determined on the basis of the cell concentration measured by the capacitance probe. Control of mu was improved by using updated information on the cell concentration compared with the simple feed-forward determination method using the initial cell concentration only. The highest specific pigment production rate was achieved with a mu of 0.02 h(-1) in the feeding phase. However, among several fermentation examined, the largest pigment production in the final step was obtained at a mu of 0.01 h(-1); in each case the same amount of substrates was used. An investigation of the optimal initial glucose concentration revealed that pigment production was maximum when the initial glucose concentration in the batch mode was 10 g/l and mu was 0.01 h(-1) in the fed-batch mode. It was also found that the pellet weight in the fermentation could be accurately estimated by image analysis. The ratio of the mycelium weight to the total cell weight estimated from information on the total cell weight and the estimated pellet weight was found to be more than 80%. However, no clear quantitative relationship could be discerned between the specific pigment production rate, rho, and the ratio of mycelium in the cell population.
Journal Article↗
Tissue inhibitor of metalloproteinases-1, matrix metalloproteinases-1 and -8, and collagenase activity levels in peri-implant crevicular fluid after implantation.
The integrity of connective tissues surrounding dental implants may be influenced by a balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The purpose of this study was to provide an overall assessment of TIMP-1, MMP-1 and -8 levels as well as collagenase activities during the wound healing process after implantation and in peri-implantitis lesions. Peri-implant crevicular fluid (PICF) was sampled with sterile paper strips from 10 osseointegrated implants of 6 subjects. Ten implants from 6 patients affected with peri-implantitis were also assessed. Gingival crevicular fluid (GCF) from 11 periodontitis-affected patients and 10 healthy volunteers served as controls. TIMP-1 and MMP-1 and -8 protein levels in the PICF were measured by ELISA, and active and APMA-activatable collagenase activities were determined by functional assays using image-analysis after SDS-PAGE. The experiment showed a significant increase in the TIMP-1 level at 1 week after implantation as compared with that in GCF from healthy periodontium. Four weeks after implantation it had reached the same level as that in the GCF of healthy subjects. The data has also disclosed a higher post-implantation collagenase activity level at 1 week than at weeks 2, 4, and 12. This may be due to the increase in MMP-1 and -8. Furthermore, peri-implantitis and periodontitis were shown to be similar inflammatory lesions in respect to MMP-1 and -8 and collagenase activities, even though the TIMP-1/MMP-1 + MMP-8 ratio was significantly lower in peri-implantitis than in periodontitis. In conclusion, the overproduction of TIMP-1 in the wound area after implantation could, to some extent, inhibit excessive tissue destruction and degradation of the neo-matrix in wound repair due to MMPs.
Adult↗
The liver in collagen diseases: pathologic study of 160 cases with particular reference to hepatic arteritis, primary biliary cirrhosis, autoimmune hepatitis and nodular regenerative hyperplasia of the liver.
AIMS/BACKGROUND: Among patients with collagen diseases, liver enzyme abnormalities are a relatively common phenomenon. To establish the liver pathology in collagen diseases, detailed pathologic studies were performed on the hepatic diseases in many patients, including various kinds of collagen diseases. METHODS: The livers from 160 patients (120 autopsy and 40 liver biopsy patients) were examined pathologically: 73 with systemic lupus erythematosus (SLE), 32 with rheumatoid arthritis (RA), 18 with polymyositis and dermatomyositis (PM and DM), 15 with systemic sclerosis (SSc), 11 with mixed connective tissue disease (MCTD) and 11 with polyarteritis nodosa (PAN). RESULTS: Liver diseases were divided into three groups: hepatic arteritis, liver diseases associated with collagen diseases (primary biliary cirrhosis, PBC; autoimmune hepatitis, AIH; nodular regenerative hyperplasia of the liver, NRH) and other liver diseases. Hepatic arteritis presenting the features of the PAN type of necrotizing arteritis was found in 27 autopsy patients. The incidence of arteritis in autopsy patients was 100% in PAN and 8.3-25% in other collagen diseases. Primary biliary cirrhosis was observed in 9 patients, 7 of whom (3 with SSc, 2 with RA, 1 with PM and DM, and 1 with MCTD) had antimitochondrial antibodies (AMA)-positive PBC, and 2 SLE patients had AMA-negative PBC. Three patients (2 with SLE and 1 with MCTD) were diagnosed clinicopathologically as having AIH. However, 3 patients (1 with SLE, 1 with MCTD and 1 with PM and DM) with clinical, biochemical and serologic data indicating probable AIH were excluded from the group with AIH association because of the liver histology (no characteristic features of AIH) and clinical course. These results indicated that data without histologic assessments of the liver are not adequate for diagnosing AIH in collagen diseases. Nodular regenerative hyperplasia of the liver was observed in 7 patients (5 with SLE, 1 with SSc and 1 with PAN). CONCLUSION: The present study offers data that are useful for the diagnosis and treatment of patients with collagen diseases and liver abnormalities.
Adolescent↗
Expression of transglutaminase activity in developing human epidermis.
Epidermal transglutaminase (TGase) is known to catalyse cross-linking of several precursor proteins in the formation of cornified cell envelope at the terminal differentiation of keratinocytes. Expression of TGase activity was studied using an in situ TGase activity assay in human fetal skin samples of 49-163 days estimated gestational age (EGA). In the early two-layered epidermis (49-56 days EGA), in situ TGase activity was not observed in the periderm cells or the basal cells. In the late two-layered epidermis (57-65 days EGA), in situ TGase activity became weakly positive in the periderm cells, but not in the basal cells. In the three-layered (66-95 days EGA) and in four- or more layered (96-135 days EGA) stratified epidermis, in situ TGase activity was still restricted only to the periderm cells. After keratinization occurred in the interfollicular epidermis (163 days EGA), in situ TGase activity was expressed in the granular and cornified layers. This unique localization of TGase activity further support the hypothesis that periderm cells form cornified cell envelope during their regression process in human fetal skin development.
Epidermis↗
Dissociation of intra- and extracellular domains of desmosomal cadherins and E-cadherin in Hailey-Hailey disease and Darier's disease.
In order to clarify the pathomechanism of acantholysis in Hailey-Hailey disease (HHD) and Darier's disease (DD), the distribution of desmosomal and adherens junction-associated proteins was studied in the skin of patients with HHD (n = 4) and DD (n = 3). Domain-specific antibodies were used to determine the cellular localization of the desmosomal transmembrane glycoproteins (desmogleins 1 and 3 and desmocollin), desmosomal plaque proteins (desmoplakin, plakophilin and plakoglobin) and adherens junction-associated proteins (E-cadherin, alpha-catenin, beta-catenin and actin). A significant difference in staining patterns between intra- and extracellular domains of desmosomal cadherins and E-cadherin was demonstrated in acantholytic cells in both HHD and DD, but not in those in pemphigus vulgaris and pemphigus foliaceus samples used as controls. In acantholytic cells in HHD and DD, antibodies against attachment plaque proteins and intracellular epitopes of desmosomal cadherins exhibited diffuse cytoplasmic staining, whereas markedly reduced staining was observed with antibodies against extracellular epitopes of the desmogleins. Similarly, membrane staining of an intracellular epitope of E-cadherin was preserved, while immunoreactivity of an extracellular epitope of E-cadherin was destroyed. While the DD gene has been identified as ATP2A2, the gene for HHD has not been clarified. The dissociation of intra- and extracellular domains of desmosomal cadherin and E-cadherin is characteristic of the acantholytic cells in HHD and DD, and not of pemphigus. This common phenomenon in HHD and DD might be closely related to the pathophysiological mechanisms in both conditions.
Acantholysis↗