High-Aspect-Ratio Ridge Structures Induced by Plastic Deformation as a Novel Microfabrication Technique.

Wrinkles on thin film/elastomer bilayer systems provide functional surfaces. The aspect ratio of these wrinkles is critical to their functionality. Much effort has been dedicated to creating high-aspect-ratio structures on the surface of bilayer systems. A highly prestretched elastomer attached to a thin film has recently been shown to form a high-aspect-ratio structure, called a ridge structure, due to a large strain induced in the elastomer. However, the prestretch requirements of the elastomer during thin film attachment are not compatible with conventional thin film deposition methods, such as spin coating, dip coating, and chemical vapor deposition (CVD). Thus, the fabrication method is complex, and ridge structure formation is limited to planar surfaces. This paper presents a new and simple method for constructing ridge structures on a nonplanar surface using a plastic thin film/elastomer bilayer system. A plastic thin film is attached to a stress-free elastomer, and the resulting bilayer system is highly stretched one- or two-dimensionally. Upon the release of the stretch load, the deformation of the elastomer is reversible, while the plastically deformed thin film stays elongated. The combination of the length mismatch and the large strain induced in the elastomer generates ridge structures. The morphology of the plastic thin film/elastomer bilayer system is experimentally studied by varying the physical parameters, and the functionality and the applicability to a nonplanar surface are demonstrated. Finally, we simulate the effect of plasticity on morphology. This study presents a new technique for generating microscale high-aspect-ratio structures and its potential for functional surfaces.

Port-site transversus abdominis fascia closure reduced the incidence of incisional hernia following retroperitoneal laparoscopic nephrectomy.

PURPOSE: The incidence of incisional hernia after laparoscopic surgery is reportedly 0-5.2 %; there are only a few reports of that following retroperitoneal laparoscopic nephrectomy. We evaluated the incidence of and risk factors for incisional hernia after retroperitoneal laparoscopic nephrectomy, and the efficacy of our novel prophylaxis technique. METHODS: A total of 207 renal cell carcinoma patients who underwent laparoscopic nephrectomy at Chiba University Hospital were retrospectively enrolled in this study. We compared the incidences of incisional hernia following the transperitoneal vs. retroperitoneal approaches, and, among the latter group, the incidences with vs. without use of our prophylaxis method. Also among the retroperitoneal-approach group, we evaluated selected patient characteristics as potential hernia risk factors. RESULTS: The rate of incisional hernias was 14 (8.7 %) after 161 retroperitoneal laparoscopic nephrectomies and one (2.2 %) after 46 transperitoneal laparoscopic nephrectomies (P = 0.132). For those undergoing the retroperitoneal approach, 14 (11.3 %) hernias were identified in 124 non-prophylaxed patients and none in 37 prophylaxed patients. Transversus abdominis fascia closure was a statistically significant factor for reducing the incidence of incisional hernia after retroperitoneal laparoscopic nephrectomy (P = 0.0324): rectus abdominis muscle thickness ≤7 mm and perioperative blood loss >100 ml were statistically significant independent risk factors, by multivariate analysis. CONCLUSIONS: To prevent incisional hernia after retroperitoneal laparoscopic nephrectomy in the patients with risk factors, it is useful to close the transversus abdominis fascia at the port sites from inside the surgical cavity, through the open specimen-removal trocar port site, under direct observation.

Clinical trial of acetazolamide in SCA6, with assessment using the Ataxia Rating Scale and body stabilometry.

OBJECTIVE: To investigate the effect of acetazolamide on spinocerebellar ataxia type 6 (SCA6). METHODS: Acetazolamide (250-500 mg/day) was administered orally for 88 weeks to 6 patients with SCA6, and its effect was quantitatively monitored using the Ataxia Rating Scale (ARS) and body sway analysis by stabilometry. RESULTS: During administration of acetazolamide, the ARS score and the amplitude of body sway were significantly reduced compared with before administration. However, the response became weaker after 1 year of treatment. CONCLUSION: Although this was an open trial, the results suggested that acetazolamide can temporarily reduce the severity of symptoms during the progression of SCA6.

Elevated parasympathetic nerve tone in isoproterenol-induced neurally mediated syncope during head-up tilt testing.

To clarify the autonomic nerve mechanisms whereby isoproterenol induces vasovagal reaction, 32 patients with recurrent syncopal episodes of unknown origin underwent head-up tilt testing. Syncope was induced in 11 patients by a control tilt (control/+ group). The remaining 21 patients (control/- group) underwent the procedure during isoproterenol infusion. Syncope was induced in 13 (isoproterenol/+ group), but not in the remaining 8 patients (isoproterenol/- group). High-frequency component (HF) and low-frequency component (LF)/HF, which are indices of heart rate variability, were assessed for 2-min intervals at baseline (T1) during the initial phase (T2) and at the end (T3) of tilting in each test. The ratios of HF and LF/HF at T2/T1 and T3/T2 were calculated. The HF value at T3 in the control/+ group was greater than that in the control/- group (1.47+/-0.91 vs 0.75+/-0.34; p<0.05). The HF value at T3 in the isoproterenol/+ group was significantly greater than those in the isoproterenol/- group (1.19+/-1.04 vs 0.43+/-0.23; p<0.05). The HF value at T2 in the isoproterenol/+ tended to be greater than in the isoproterenol/- group (1.30+/-0.85 vs 0.66+/-0.53; p=0.07). The ratio of HF for T2/T1 in isoproterenol/+ was greater than in the other groups (2.04+/-1.26 in isoproterenol/+ vs 0.58+/-0.39 in control/-, 0.47+/-0.18 in control/+ and 0.71+/-0.45 in isoproterenol/-; p=0.0001), whereas that of HF for T3/T2 in the control/+ group was greater than in the other groups (1.48+/-0.62 in control/+ vs 0.97+/-0.33 in control/-; p<0.05, 0.79+/-0.35 in isoproterenol/- and 0.88+/-0.37 in isoproterenol/+; p<0.01). No difference was observed in either LF/HF or the ratio of LF/HF among the groups. The early and persistent increase in parasympathetic nerve tone provoked by isoproterenol plays an important role in enhancing the vasovagal reaction during head-up tilt testing.

Expression of adhesion molecules by human endothelial cells exposed to oxidized low density lipoprotein. Influences of degree of oxidation and location of oxidized LDL.

The main objective of this study was to determine the influence of the degree of low density lipoprotein (LDL) oxidation and the location of oxidized LDL (oxLDL) on expression of adhesion molecules on endothelial cells (EC). OxLDL preparations 1-4 with different degrees of oxidative modification were studied. All preparations of oxLDL, after addition to the medium, stimulated the expression of intercellular adhesion molecule-1 (ICAM-1) by human umbilical vein endothelial cells (HUVEC) as determined by cell-ELISA. Concentration-dependent studies examining ICAM-1 expression by HUVEC showed that the minimal concentration of oxLDL which significantly stimulated ICAM-1 expression was 5 microg/ml, suggesting that the predicted physiological concentration of oxLDL in plasma may be not high enough to elicit a substantial increase of ICAM-1 expression in EC. In contrast, very small amounts (0.15 microg/well) of oxLDL-3 and 4, the more heavily oxidized LDL preparations, stimulated effectively ICAM-1 expression by HUVEC when located below the endothelial cell monolayer by immobilizing to type I collagen. The results suggest that the increased expression of ICAM-1 induced by accumulated oxLDL may be one of the mechanisms by which oxLDL contributes to atherogenesis.

[Results of two-step tuberculin skin test against medical and nursing school students and treatment according to the results of the test].

To assess the risk of tuberculosis infection in medical and nursing school students, tuberculin skin tests were carried out in the two-step manner. The second tuberculin skin test was repeated two weeks later excluding those who were strongly positive in the first test. BCG vaccination was done with the consent of students who showed negative reaction twice. Medical interview and revaluation of prior routine chest radiogram were made on students who were strongly positive. Prophylactic INH medication was considered to those who are at high risk of tuberculosis. Eight hundred thirty eight students underwent the two-step tuberculin skin test, and among them, 771 students showed the positive reaction on the first test (92.0%) which included 58 weakly positive (6.9%), 347 intermediately positive (41.4%) and 366 strongly positive (43.7%) and 2 not-measurable (0.2%), and 65 students were negative (7.8%). The average size of the erythema was 30.9 +/- 18.8 mm on the first test and 37.9 +/- 20.6 mm on the second test. Twenty one students were negative on the second tuberculin skin test, and among them, 15 received BCG vaccination. Out of eight students who were vaccinated with BCG in 1999 and were followed up in the next year, 6 (75.0%) converted to positive. Strongly positive reaction was seen in 28 students (3.3%) and one of them underwent prophylactic medication of INH according to her family history of exposure to tuberculosis.

Correlation of magnetic resonance imaging and histological findings in a large basilar tip aneurysm after coil embolization--case report.

A 73-year-old female with a large basilar tip aneurysm underwent endovascular coil embolization with interlocking detachable coils (IDCs). The patient subsequently died of pneumonia 25 days after the embolization, and the aneurysm specimen was obtained at autopsy. Histological examination showed that the intraaneurysmal structure consisted of three layers. The outer layer was mildly organized thrombus surrounding the coils, the middle layer was disorganized clot, and the inner layer consisted of fresh blood clot. Gradient-echo magnetic resonance (MR) imaging taken before death had demonstrated a central region of high intensity and a peripheral low intensity region corresponding to the inner and middle-outer layers of the aneurysm, respectively. Intraaneurysmal placement of IDCs leads to the formation of a clot surrounding the coils. However, clot formation may be inadequate where the packing of the IDCs is incomplete. Comparison of the MR imaging and histological findings in this case show that gradient-echo MR imaging can assess thrombus and residual blood flow within the aneurysm.

A Case Report of Neovascular Glaucoma Due to Carotid Artery Occlusive Disease. Diagnosis of Neovascular Glaucoma and Histological Characteristics.

Purpose: The methods of diagnosing neovascular glaucoma were evaluated in a case with carotid artery occlusive disease. A trabeculectomy specimen taken from this patient was examined histologically to find the characteristic picture of this slowly progressive neovascular glaucoma.Materials and Methods: The left eye of a 48-year old man with neovascular glaucoma due to carotid artery occlusive disease had been previously diagnosed as uveitis with ocular hypertension. His left eye was examined by fluorescein fundus and gonioangiography. Trabeculectomy was performed because of uncontrolled intraocular pressure and the trabeculectomy specimen was examined histologically including immunohistochemistry with antibodies against von Willebrand factor antigen.Results: The characteristic picture of fluorescein fundus angiography was filling delay in the choroid and stained walls of the retinal arteries and veins. Fluorescein gonioangiography using a fundus camera clearly demonstrated new vessels in the pupil and angle with intense fluorescein leak in spite of faint neovascularization in those areas when observed by slit lamp with gonioscopy. Histological examination of the trabeculectomy specimen revealed proliferation of new vessels in the spaces of the trabecular meshwork and focal occlusion of Schlemm's canal.Conclusion: Fluorescein fundus angiography and gonioangiography by fundus camera is very useful for diagnosis of neovascular glaucoma due to carotid artery occlusive disease. Neovascular proliferation in the trabecular meshwork with slight of peripheral anterior synechia and focal occlusion of Schlemm's canal may be peculiar to such slowly progressive neovascular glaucoma.

[A case report of neovascular glaucoma due to carotid artery occlusive disease--diagnosis of neovascular glaucoma and histological characteristics].

PURPOSE: The methods of diagnosing neovascular glaucoma were evaluated in a case with carotid artery occlusive disease. A trabeculectomy specimen taken from this patient was examined histologically to find the characteristic picture of this slowly progressive neovascular glaucoma. MATERIALS AND METHODS: The left eye of a 48-year old man with neovascular glaucoma due to carotid artery occlusive disease had been previously diagnosed as uveitis with ocular hypertension. His left eye was examined by fluorescein fundus and gonioangiography. Trabeculectomy was performed because of uncontrolled intraocular pressure and the trabeculectomy specimen was examined histologically including immunohistochemistry with antibodies against von Willebrand factor antigen. RESULTS: The characteristic picture of fluorescein fundus angiography was filling delay in the choroid and stained walls of the retinal arteries and veins. Fluorescein gonioangiography using a fundus camera clearly demonstrated new vessels in the pupil and angle with intense fluorescein leak in spite of faint neovascularization in those areas when observed by slit lamp with gonioscopy. Histological examination of the trabeculectomy specimen revealed proliferation of new vessels in the spaces of the trabecular meshwork and focal occlusion of Schlemm's canal. CONCLUSION: Fluorescein fundus angiography and gonioangiography by fundus camera is very useful for diagnosis of neovascular glaucoma due to carotid artery occlusive disease. Neovascular proliferation in the trabecular meshwork with slight of peripheral anterior synechia and focal occlusion of Schlemm's canal may be peculiar to such slowly progressive neovascular glaucoma.

[Effective melatonin therapy in a case of Machado-Joseph disease with insomnia].

The effects of melatonin (MLT) therapy on insomnia and serum MLT concentration were studied in a 44-year-old man with Machado-Joseph disease. He suffered from insomia since age 41, and could not sleep well, despite that he was given many kinds of hypnotics. He showed neither noctural central apnea, inspirative stridor, restless leg syndrome, REM without atonia, nor noctural polyuria, all of which are known to cause insomnia in spinocerebellar degeneration. The peak serum level of MLT was low, 9.2 pg/ml. He was administered 9 mg of MLT p.o. at 8 p.m., and it's peak noctural level elevated to 7,000 pg/ml, and this alleviated his intractable insomnia. Our experience indicate that the MLT could be effective for insomnia in MJD which relates to low MLT concentration.

[A clinical trial of acetazolamide for SCA6].

Spinocerebellar ataxia type 6 (SCA 6) is an allelic disorder of episodic ataxia type 2 (EA 2) and is caused by a small CAG repeat expansion in the gene encoding the alpha 1A-voltage-dependent-Ca channel subunit (CACNA 1 A) on chromosome 19p13.1. The disorder starts at adulthood with progressive cerebellar ataxia, and the symptoms often fluctuate at early stage. These clinical features overlap with those of EA 2, which has been known as acetazolamide-responsive ataxia. On this background, we studied the clinical effectiveness of acetazolamide for SCA 6 in 9 consecutive patients. Their clinical severity was serially evaluated by ARS (ataxia rating scale) and gravimetric test, over 32 weeks of oral administration of acetazolamide (250-500 mg/day). Consequently, a significant improvement was observed in ARS and postural sway. Our results indicate that acetazolamide is temporally effective for ameliorating the symptoms of SCA 6. However, its effects for the disease progression need to be examined in more large scales in number and duration.

Determinant of microvolt-level T-wave alternans in patients with dilated cardiomyopathy.

OBJECTIVES: The aim of this study was to clarify the clinical significance and the determinant of microvolt-level T-wave alternans (TWA) in patients with dilated cardiomyopathy (DCM). BACKGROUND: The prevention of sudden death in patients with DCM remains the therapeutic target. T-wave alternans has been proposed as a powerful tool for identification of patients at high risk for ventricular arrhythmias and sudden death in coronary artery disease. METHODS: In 58 DCM patients, TWA was measured during bicycle exercise testing using a CH 2000 system (Cambridge Heart, Bedford, Massachusetts). The New York Heart Association class, signal-averaged electrocardiogram, QT dispersion, left ventricular end-diastolic diameter (LVDd) and percent fractional shortening detected by echocardiogram and the grade of the ventricular arrhythmia were obtained in all patients. RESULTS: T-wave alternans was positive in 23 patients (TWA+ group), negative in 25 (TWA- group) and indeterminate in 10. Univariate analysis showed that the percentage of patients with ventricular tachycardia (VT) and the LVDd in the TWA+ group was significantly higher than those in the TWA- group (61% vs. 8%, p < 0.001 and 65 +/- 11 mm vs. 58 +/- 8 mm, p < 0.05, respectively). The sensitivity, specificity and predictive accuracy of TWA for VT were 88%, 72% and 77%, respectively. Multivariate analysis showed that the presence of VT was a major independent determinant of TWA in patients with DCM (p = 0.003). CONCLUSIONS: T-wave alternans was closely related to VT in patients with DCM. T-wave alternans is a useful noninvasive test for identifying high risk patients with DCM who have VT.

Oxidized LDL-containing immune complexes induce Fc gamma receptor I-mediated mitogen-activated protein kinase activation in THP-1 macrophages.

Our previous studies have shown that Fc gamma receptor (FcgammaR)-mediated uptake of LDL-containing immune complexes (oxLDL-ICs) by human monocyte-derived macrophages leads to not only transformation of macrophages into foam cells but also macrophage activation and release of cytokines. It has been shown that cross-linking of FcgammaR triggers activation of signal transduction pathways that alter gene expression in macrophages. In this study, we determined whether engagement of FcgammaR by oxLDL-ICs leads to activation of mitogen-activated protein (MAP) kinase pathway, a signaling cascade serving many important functions, including the regulation of gene expression, in THP-1 macrophage-like cells. Our results from immunoblotting, using specific anti-phosphorylated MAP kinase antibodies, showed that oxLDL-ICs induced extracellular signal regulated kinase 2 (ERK2) MAP kinase phosphorylation in THP-1 macrophage-like cells in time- and dose-dependent manners. Cholesterol loading before stimulation led to a longer phosphorylation of ERK2. Nuclear translocation of phosphorylated ERK was markedly increased after the stimulation. Moreover, our data showed that oxLDL-IC induction of MAP kinase was prevented by human monomeric IgG1, suggesting that the specific engagement of type I FcgammaR by oxLDL-IC is responsible for the MAP kinase activation. Finally, we showed that human anti-oxLDL autoantibody-containing immune complexes immobilized on type I collagen induced MAP kinase activation in THP-1 cells. These results strongly suggest that oxLDL-IC, which has been detected in atherosclerotic plaques, may play an important role in macrophage activation and atherogenesis.

Monkey hepatocytes efficiently express tissue factor pathway inhibitor (TFPI), in contrast with human and rat hepatocytes.

It has been reported that tissue factor pathway inhibitor (TFPI), a Kunitz-type protease inhibitor that regulates the extrinsic blood coagulation pathway, is not expressed in human, bovine, rabbit, or rat liver. Here, we found that TFPI is efficiently expressed in Macaque monkey liver. Monkey hepatocytes were identified as the expression cells by Northern blot analysis. The hepatocytes were stained with anti-human TFPI antibody, as were endothelial cells of the small vessels. We isolated and sequenced the 5'-flanking 1.4 kb regions of monkey and human TFPI genes, and found them to show 92.6% identity in their nucleotide sequences. We measured their transcriptional activities using a luciferase reporter gene and showed that the activity of the monkey TFPI gene is higher than that of the human gene in monkey primary hepatocytes. Although the binding motif of hepatocyte nuclear factor-1 is present only in the monkey gene, the site does not seem to be involved in the transcriptional activity. Mutagenetic analyses revealed that the region from -138 to +28 in the monkey gene is important for the expression of TFPI in hepatocytes. The present study indicates that the expression of the monkey TFPI gene is regulated by different mechanisms from the human TFPI gene.

Fluorescence automatic cell sorter and immunohistochemical investigation of CD68-positive cells in meningioma.

Infiltration of brain neoplasms by mononuclear cells including monocytes/macrophages has attracted little attention since they have marked morphological heterogeneity. Twenty-seven meningiomas were studied by anti-CD68 antibody-gated flow cytometry and by immunohistochemical analysis using the anti-CD68 antibodies. Flow cytometric analysis divided cells contained within tumor tissues into CD68-positive and -negative cells. In addition, eight gliomas, eight metastatic brain tumor, and 12 pituitary adenomas were investigated in the same way to compare meningiomas. The mean contents of CD68-positive cells were 24.0 +/- 3.7% in meningiomas, 4.4 +/- 1.4% in gliomas, 9.5 +/- 3.9% in metastatic brain tumors, and 4.5 +/- 1.8% in pituitary adenomas. Immunohistochemically, CD68-positive cells showed significant heterogeneity and were detected as round, rod-shaped, ameboid and ramified cells in meningiomas. Although the infiltrated mononuclear cells in gliomas have been investigated to some degree and showed that they express cytokines and/or growth factors, these infiltrated cells in meningioma have barely been studied. The CD68-positive cells detected in this study are likely to be monocytes, macrophages and microglias, and are presumed to be in various functional stages and to play important roles in growth regulation in meningioma.

[Initial symptoms and mode of neurological progression in spinocerebellar ataxia type 6 (SCA6)].

Spinocerebellar ataxia type 6 (SCA6) is genetically defined as a group of SCA characterized by late-onset pure cerebellar ataxia clinically and by a small CAG repeat expansion in the gene encoding the alpha 1A-voltage-dependent-Ca channel subunit (CACNL1A4) on chromosome 19p13.1 genetically. We analyzed the initial symptoms and the mode of progression in this disorder on 25 genetically verified patients. The initial symptoms were recurrent episodes of transient vertigo (72%) or unsteady gait (28%). Neurologically, they showed apparent gaze-evoked nystagmus (92%), transient positional nystagmus (83%), and periodic alternating nystagmus (4%), in addition to cerebellar ataxia. In addition to these episodic symptoms, all patients developed progressive cerebellar ataxia over years. These fluctuating symptoms at the initial stage of the illness were clearly different from those of other SCA, rather overlapping with those of episodic ataxia type 2 (EA2), an allelic disorder of SCA6. The clinical similarity indicates that there might be a common mechanism at least in part causing these two disorders. The mode of progression and their neurological features were also presented.

The relation between the pacing sites in the right ventricular outflow tract and QRS morphology in the 12-lead ECG.

Optimal pace mapping is a good predictor of the appropriate ablation site for idiopathic right ventricular tachycardia (VT) originating from the right ventricular outflow tract (RVOT). We studied the relationship between the RVOT pacing site and QRS morphology in the 12-lead ECG during pacing to find the optimal site more quickly. In 13 patients with idiopathic VT, pacing at 8 sites in the RVOT (free wall, septum, and anterior and posterior regions of upper and lower sites) was performed while 12-lead ECGs were recorded. The R-wave amplitude minus the S-wave amplitude in lead I (RI-SI) and aVF (RavF-SavF) and the transitional zone index (TZI) were compared in the different pacing sites; TZI was defined to examine the transitional zone as a value. The RI-SI was smaller in the anterior region than in the other regions, and the negative RI-SI predicted that the pacing site was in the upper or lower anterior regions. The RavF-SavF was larger in the septum and anterior regions than in the free wall and posterior regions. The TZI was larger in the free wall region than in the septum. From the results, we constructed a flow chart that differentiates the origin of the arrhythmia in the RVOT and a directional guide that indicates the direction from the current mapping site for optimal pace mapping. The results provided an ECG guide for locating the focus of VT originating from the RVOT. These findings may systematically improve the mapping procedure.