[Microquantitative analysis of serum haptoglobin by agar gel electrophoresis].
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Biomedical subjects
Publications and source records attributed to M Arai.
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In corneal scraping smears from 13 patients with clinically suspected herpes simplex keratitis (HSK), HSK is demonstrated by means of peroxidase-antiperoxidase (PAP) technique with antisera to herpes simplex virus (HSV) in Papanicolaou-destained cellular samples. The staining for HSV antigen was present in seven cases of corneal scraping smears with superficial keratitis (dendritic and geographic ulcers) while six cases of stromal keratitis (deep keratitis) failed to show HSV antigen except in one case. Specific antigen for HSV was predominantly present in the cytoplasm rather than in the nucleus. Immunoreactions were negative with HSV antisera in patients with other infections and in those in a normal control group. Using the PAP technique, detection of HSV antigen in corneal scraping smears was of great value in the diagnosis of HSK, especially in cases of superficial keratitis.
Baboons fed ethanol (50% of total calories) chronically develop ultrastructural alterations of hepatic mitochondria. To determine whether mitochondrial functions are also altered, mitochondria were isolated from nine baboons fed ethanol chronically and their pair-fed controls. At the fatty liver stage, ADP-stimulated respiration was depressed in ethanol-fed baboons by 59.4% with glutamate, 43.2% with acetaldehyde, 45.1% with succinate and 51.1% with ascorbate as substrates. A similar decrease was noted in the ADP/O ratio (14 to 28%) and respiratory control ratio (20 to 44%) with all substrates. Similar alterations of mitochondrial functions were observed in baboons with more advanced stages of liver disease, namely fibrosis. These changes after ethanol treatment were associated with decreases in the enzyme activities of mitochondrial respiratory chain: glutamate, NADH and succinate dehydrogenase (42, 24 and 28%, respectively), glutamate-, NADH- or succinate-cytochrome c reductase (42, 27 and 32%, respectively) and cytochrome oxidase (59.6%). The content of all cytochromes was also decreased in ethanol-fed baboons, especially aa3 (57%). Moreover, [14C]leucine incorporation into mitochondrial membranes was depressed by 21% after ethanol treatment. On the other hand, glutamate dehydrogenase activities of serum and cytosol in ethanol-fed baboons were significantly higher than those in pair-fed controls. Morphologically, mitochondria of ethanol-fed baboons were larger than those of pair-fed controls. However, the mitochondrial protein content per mitochondrial DNA was unchanged. From these results, we conclude that, morphologically and functionally, hepatic mitochondria in baboons are altered by chronic ethanol consumption; it is noteworthy that these changes are fully developed already at the fatty liver stage, and that morphological alteration appears to reflect the damage of mitochondrial membranes rather than an adaptive hypertrophy.
Laparoscopic cholecystectomy has represented a potentially more morbid procedure than open cholecystectomy. Some of this morbidity has been due to complications associated with pneumoperitoneum. We have developed a technique that employs abdominal wall retraction during laparoscopic cholecystectomy and allows access to the right upper part of the abdomen without maintenance of pneumoperitoneum. Among 151 patients who underwent laparoscopic cholecystectomy using abdominal wall retraction there were no recognized adverse effects. Abdominal wall retraction enables the surgeon to minimize the risk of serious complications associated with pneumoperitoneum during laparoscopic cholecystectomy.
Oral administration of CS-500, a competitive inhibitor of 3-hydroxy-3-methylglutaryl CoA reductase, lowered the serum cholesterol level of both normal and WHHL-rabbits (the first example of heritable hyperlipidemic animals) at doses higher than 5 mg/kg/day. Phospholipids decreased concomitantly, whereas triglycerides did not in either normal or WHHL-rabbits.
Serum activity of angiotensin converting enzyme (ACE) was serially measured in 47 hospitalized chronic alcoholics with liver disease. Compared to healthy controls, ACE activity, on admission, in the serum of alcoholics was significantly elevated (42.5 +/- 16.6 U/ml vs. 32.4 +/- 9.6 U/ml; p less than 0.005). About 36% of the patients had an elevated ACE level exceeding an upper normal value of 42 U/ml (mean +/- SD). In contrast to the rapid normalization of such enzymes as aspartate transaminase (AST), alanine transaminase (ALT) and lactic dehydrogenase (LDH) which represent parenchymal liver cell injury, the activity of ACE remained elevated over a period of 4 weeks even with abstinence. The serum level of ACE was significantly correlated with levels of alkaline phosphatase, gamma-glutamyltranspeptidase and monoamine oxidase, but not with those of AST, ALT and LDH. These data suggest increased ACE activity in alcoholics may be related to the influence of chronic consumption of alcohol on hepatic nonparenchymal systems.
In order to clarify the origin and the mechanism of increased serum activity of glutamic oxalacetic transaminase (GOT) in chronic alcoholics, clinical and experimental investigations were carried out. Mitochondrial (m-GOT) and cytosolic GOT (c-GOT) isoenzymes were separated chromatographically by using a mini-column packed with Sephadex A50. Sixty percent of 63 alcoholics had elevated serum GOT. The m-GOT activity in alcoholics with total serum GOT activity of over 50 Karmen Units was 17.2 +/- 1.6 K.U. and the m-GOT/GOT ratio was the highest when compared to those in non-alcoholic liver diseases. In in vitro study, six hours of incubation of isolated hepatocytes from rats fed ethanol chronically resulted in an increased leakage of m-GOT into the incubation medium and also showed a tendency of a higher m-GOT/GOT ratio than that from control rats. The m-GOT activity thus released into the medium showed a highly significant inverse correlation with the viability of hepatocytes. These data suggest that m-GOT substantially contributes to an increased serum GOT often observed in chronic alcoholics.
Multifocal electroretinography (ERG), developed by Sutter in 1992, is a method of recording the spatial distribution of focal ERG in a short time period using multi-input stimulation. Using this technique, we can detect the spatial extent and severity of damage to the macula. In this study, we recorded multifocal ERGs from 20 eyes of 20 normal subjects and analyzed the topographical properties of responses. In every subject, a negative wave followed by a positive wave could be recorded and we named them the N1-wave and the P1-wave, respectively. The amplitudes of the N1-wave and the P1-wave were the largest in the fovea, and they became smaller with eccentricity. In the P1-wave amplitude, the greatest intersubject variability was observed at the fovea. The N1 and P1 latencies were shorter in the upper retina than in the lower retina. The amplitude was larger in the upper retina than in the lower retina, which suggests the functional superiority of the upper retina. There was no statistical difference in latency and amplitude between the nasal and the temporal retina. We found no statistical difference between the responses of the papillomacular bundle and those of the temporal retinal area. The mapping obtained by multifocal ERG was useful as objective perimetry.
BACKGROUND: Voluntary nystagmus has been recognized as a pendular, rapid, conjugate, primarily horizontal, benign eye movement initiated and maintained by voluntary effort. CASE: A 10-year-old Japanese girl presented with voluntary nystagmus associated with accommodation spasms. Her chief complaints, intermittent blurred vision, headache, and soreness of the eyes, were thought to be related to the voluntary nystagmus and accommodation spasms. FINDINGS: The waveform of the nystagmus appeared pendular, the frequency was 13-15 Hz, and the amplitude was 3-5 degrees. Scanning laser ophthalmoscopic video images clearly demonstrated vertical and torsional components in addition to the horizontal eye movements. Her refraction was unstable, varying between -0.5 diopters (D) and -5.5 D, and the recording of the accommodometer increased to -12.0 D when nystagmus was initiated. CONCLUSIONS: This may be a unique form of voluntary nystagmus that consists of horizontal, vertical, and rotational components associated with accommodation spasms. Observation of this patient continues, without any further treatment or examination.
PURPOSE: To investigate how the multifocal electroretinogram (ERG) is altered in conditions of blocking, light scattering, or distortion of the stimulus that are seen in ocular pathologies. METHODS: A central 40 degree-diameter stimulus pattern consisting of 61 hexagons was presented on a cathode ray tube monitor at a rate of 75 Hz according to the pseudo-random binary M sequence by the Veris computer program. Localized responses corresponding to each hexagon and ERG topographies were displayed on the computer screen. Central scotoma was simulated by blocking the central area of the stimulus, visual field constriction by blocking the outer area of the stimulus, mild cataract by using acrylic filters that caused light scatter, and epiretinal membrane by using a wavy plastic plate that produced metamorphopsia. RESULTS: The responses from the blocked area were nonrecordable whether blockage was central or peripheral; responses from the adjacent unblocked area had a larger amplitude when large areas of the stimulus were blocked. The light scatter that decreased vision from 20/20 to 20/70 did not significantly decrease response amplitudes. Responses from areas in which the stimulus pattern was distorted were minimally affected. CONCLUSIONS: The results show that the system records local ERGs from the macula and outside the macula. It can detect the area where the stimulus is blocked. Moderate light scattering and distortion do not cause loss of local ERG characteristics.
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In the present study, the effect of chronic ethanol consumption in rats on the hepatic heme metabolism was investigated. Male Wistar rats were fed a nutritionally adequate liquid diet containing ethanol as 36% of the total calories for 5 weeks. After an overnight fast, the livers were excised and centrifuged to obtain mitochondrial and microsomal fractions. Chronic ethanol feeding of rats resulted in about 19% hepatomegaly as represented by the increased liver/body weight ratio. There was no difference in the mitochondrial protein content between the ethanol-treated and control rats, but the microsomal protein content was significantly increased in the ethanol-treated rats. Hepatic microsomal content of cytochrome P-450 (P-450) was markedly enhanced by chronic ethanol ingestion. Microsomal contents of cytochrome b5 (b5) and total heme were also increased to a lesser extent. After chronic ethanol abuse, the hepatic activity of delta-aminolevulinic acid (ALA) synthetase, which is a rate-limiting enzyme for heme production, was significantly increased and that of the heme oxygenase was slightly increased. These data indicate that ALA synthetase activity is induced by the negative feedback mechanism in order to compensate the depletion of heme caused by the utilization of heme for P-450. It is also speculated that, in response to excessive production of heme as described above, heme oxygenase activity is secondarily induced to regulate the amount of heme.
Abdominal aortic aneurysms (AAAs) are characterized by structural deterioration of aortic wall leading to progressive dilatation. The histopathological changes in AAAs are particularly evident within the elastic media, which is normally comprised mainly of vascular smooth muscle cells (SMCs). There are vascular myosin heavy chain (MHC) isoforms; SM2 is specifically expressed in differentiated SMCs and SMemb is a nonmuscle-type MHC abundantly expressed in SMCs of the fetal aorta with an immature phenotype. Although AAA altered expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), pathophysiological role of SMC phenotypic modulation in the AAA progression remains uncertain. To determine whether phenotypic modulation in vascular SMCs contributes to arterial medial degeneration, we examined MHC expression in SMCs of AAA. Aortic specimens were obtained from patients with slowly progressed AAA (n = 12) and rapidly progressed AAA (n = 5), and compared with normal aortic tissue (n = 3). Immunohistochemical staining was performed for detection of SMemb, SM2, MMP (types 2 and 9) and TIMP (types 1 and 2). Faint SMemb and abundant SM2 were observed in normal aorta, while the balance shifted to SMemb predominance in AAAs. Compared with slowly progressed AAA tissue, rapidly expanded AAA tissue demonstrated marked increases in SMemb expression with suppressed SM2. Predominant SMemb expression indicates presence of phenotypic modulated SMCs and enhanced MMP; while abundant TIMP was seen in mature SMCs expressing SM2. SMemb expression is markedly increased in AAA with MMP enhancement, and a significant imbalance between SMemb and SM2 results in rapid progression of AAA.
BACKGROUND: To evaluate cardiac sympathetic activity, a simple method should be developed to replace such complex methods as the spillover rate of tritiated norepinephrine (3H-norepinephrine) or microneurography of sympathetic nerve activity. The goal of this study is to evaluate cardiac sympathetic activities by analyzing the washout of I-123 meta-iodobenzylguanidine (123I-MIBG), a radiolabeled norepinephrine analogue, in Dahl salt-sensitive (DS) rats as it relates to the progression of hypertension. METHODS AND RESULTS: Dahl salt-resistant (DR) rats and DS rats were fed an 8 % salt diet starting at age 5 weeks. Marked hypertension and cardiac hypertrophy developed in the DS rats, whereas DR rats remained normotensive. Then the time-activity curves of 123I-MIBG from 15 to 200 minutes were obtained from both DS and DR strains at ages 8, 11, and 13 weeks using dynamic scintigraphic analysis. We also examined the nonneuronal washout of 123I-MIBG using dynamic scintigraphic studies in desipramine pretreated normal rats. In the preliminary study with desipramine pretreatment, the majority of the nonneuronal 123I-MIBG washout occurred by 90 minutes after injection. Therefore the late-phase washout in the control rats was found to reflect the neuronal washout. We then applied exponential curve fitting to the time activity curves acquired in the 90- to 200-minute period after 123I-MIBG injection in both DR and DS rats. When we compared the coefficients of these washout curves in the DS and DR rats as an index of cardiac sympathetic activities, the coefficient values remained high during all stages in DS rats, whereas they decreased with age in DR rats. CONCLUSION: Measurement of late-phase 123I-MIBG washout may be a useful tool for assessing the change in sympathetic activity in the progression of hypertension without the influence of extraneuronal washout of 123I-MIBG and left ventricular hypertrophy.
This study tested the usefulness of technetium-99m-labeled antigranulocyte monoclonal antibody BW250/183 (AGMAb) for identifying granulocyte accumulation in ischemic/reperfused canine myocardium. In dogs with 90 minutes coronary artery occlusion and 180 minutes reperfusion (n = 8), ischemic/reperfused myocardial samples demonstrated 8.5 +/- 2.4 times more Tc-99m-AGMAb accumulation than nonischemic samples. Dogs given Tc-99m-labeled nonspecific human immunoglobulin instead of Tc-99m-AGMAb (n = 3) had about half as much accumulation (4.5 +/- 1.6, P < .05). Ex vivo myocardial imaging of Tc-99m-AGMAb demonstrated marked uptake in infarcted regions identified by absent triphenyl tetrazolium chloride staining. The amount of uptake was inversely related to the severity of ischemia (determined by radioactive microspheres) and directly correlated with tissue myeloperoxidase activity, a specific marker of granulocyte accumulation. No increase in Tc-99m-AGMAb uptake occurred in dogs with 90 minutes ischemia and no reperfusion (n = 3) or 15 minutes ischemia and 180 minutes reperfusion (n = 2). In conclusion, Tc-99m-AGMAb is taken up in reperfused infarcted myocardium by both nonspecific and specific mechanisms. Because the amount of uptake reflects myocardial granulocyte accumulation, Tc-99m-AGMAb combined with nuclear imaging techniques may be useful for studying inflammatory processes in the heart in experimental animal models and human beings.
PURPOSE: To determine if the pattern reversal visual evoked response (PVER) can objectively evaluate visual function in retinitis pigmentosa (RP). METHODS: We retrospectively reviewed the medical records of 29 patients (29 eyes) with RP, a visual acuity (VA) better than 20/100, and for whom an electroretinogram (ERG) and a PVER had been recorded. A steady-state PVER was measured using five check sizes: 160, 80, 40, 20 and 10 min of arc. The best-corrected VA was measured using the ETDRS chart. Visual fields (VFs) were measured using standard Goldmann perimetry. To quantify the VFs, we measured the field size (isopter IV-2) in four meridians and averaged them. Twenty-five eyes of 25 normal subjects served as controls. RESULTS: The mean and median VAs were 20/40 (logMAR = 0.30 +/- 0.20). The average VF (isopter IV-2 ) was 20 degrees (median, 9 degrees: range. 2 degrees to 62 degrees). All patients had a recordable PVER. Only 12 (41 %) patients had a recordable ERGs, all subnormal. The PVER amplitudes showed a relatively good correlation with VA in the intermediate check sizes (40 min of arc, r = 0.611. P = 0.0004: 20 min of arc, r = 0.596. P = 0.0007). The PVER amplitude-check size function had a flattened inverted V shape in the RP patients and an inverted U shape in the normal subjects, with the mean amplitudes for the RP patients significantly smaller than the normals at all check sizes. CONCLUSION: The PVER is a useful method for objectively evaluating the visual function in RP patients whose central vision is still preserved.