Showing papers in "Tohoku Journal of Experimental Medicine in 2002"

Recent Studies on Metallothionein: Protection Against Toxicity of Heavy Metals and Oxygen Free Radicals

Abstract: Metallothionein (MT) is a ubiquitous, cysteine-rich, metal-binding protein. MT synthesis is induced by various stimuli such as cadmium, mercury, zinc, oxidative stress, glucocorticoid, and anticancer agents. Recently, transgenic mice with loss-of-function mutations in the MT-I/-II genes were established. It has been assumed that MT plays a role in the detoxification of heavy metals. In recent studies using MT-null mice, the ability of MT to protect against cadmium-induced renal, liver and bone injuries has been confirmed. Moreover, MT is also capable of scavenging oxygen free radicals. MT is involved in the protection of tissues against various forms of oxidative injury, including radiation, lipid peroxidation, oxidative stress caused by anticancer drugs, and conditions of hyperoxia. However, MT still lacks an established biological function. Unexpectedly, the MT-null mice were apparently in good health, and the critical biological roles of MT have been questioned. MT seems to be a protective protein produced in response to a variety of stresses. Here, current studies on the protective roles of MT against toxicity of heavy metals and reactive oxygen species are reviewed, and the putative biological functions of MT are discussed.

Modification of Mercury Toxicity by Selenium : Practical Importance?

Abstract: The interaction between mercury and selenium may involve a variety of toxicologically and biochemically distinct processes. In this paper, the interaction between inorganic mercury and sodium selenite, the interaction most extensively studied, as well as the interaction between methylmercury (MeHg) and selenium, the interaction perhaps most significant for non-occupational human populations, will be discussed. It has been shown that the former interaction can be understood as a modification of the kinetic behavior of inorganic mercury by selenite, but this interaction may occur only under very limited conditions. On the other hand, the mechanism of the latter interaction is largely unknown, and kinetic modification appears to play only a minor role. An interaction between MeHg and selenoproteins or a possible interaction between the inorganic mercury, resulting from the demethylation of MeHg, and the selenium may be important. Compared to the experimental findings, little evidence of the toxicological modification of MeHg by selenium was obtained in epidemiological studies.

Perspectives on cadmium toxicity research.

Abstract: Since there are a plethora of studies on cadmium toxicity and poisoning in laboratory animals and humans, we have limited this review to studies that are relevant to human health issues by focusing on carcinogenicity, genotoxicity, circulatory disease, nephrotoxicity and life expectancy. Cadmium exposure has been established to induce cancer in various tissues of laboratory animals. Contrary to early findings of the lack of genotoxicity by cadmium, recent findings of mammalian cell culture studies have revealed genotoxic effects. Furthermore, cadmium exposure at relatively low doses induces circulatory diseases in laboratory animals. Despite such results of various cadmium toxicities in animal studies, data from human studies are lacking and insufficient to support the cause-effect relationship. Although cadmium is currently considered to be a human carcinogen by the International Agency for Research and Cancer, it is inappropriate to conclude that sufficient evidence on the carcinogenicity of cadmium in humans exists. It is also thought that epidemiological studies so far reported do not support the occurrence of cadmium-induced circulatory disease in humans. Since there are inconsistent reports on the relationship of cadmium exposure with the life expectancy of people living in cadmium-polluted areas, further studies are needed for clarification. It is also necessary to examine apparent discrepancies in result between humans and experimental animals. It has been established that long-term exposure to cadmium causes renal dysfunction in both humans and experimental animals, and whether there are any differences in the inducibility of metallothionein in the kidney warrants further study.

Lipid peroxidation and antioxidant enzymes in Turkish population: relation to age, gender, exercise, and smoking.

Abstract: The purpose of this study was to examine the change in lipid peroxidation and antioxidant enzyme activities in healthy subjects and to evaluate the concentrations of superoxide dismutase, glutathione peroxidase and malondialdehyde, an end product of lipid peroxidation in exercise and smoking. Study included 257 appearently healthy individuals, 133 males and 124 females. In all subjects, malondialdehyde (MDA) levels were analyzed as an indicator of the lipid peroxidation activities. Superoxide dismutase, glutathione peroxidase activities were measured as an indicator of antioxidant activities. Oxidative stress was estimated by the method based on thiobarbituric acid reactivity. Erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were estimated on hemolysates by use of commercial available kits (Randox lab., Dublin, Ireland). For all groups serum lipid peroxidation and erythrocyte SOD and GSH-Px were obtained at the initial and the following periods. Serum MDA level was higher in the elderly than in the children and in the adults. MDA levels were higher in the smoking, acute exercise than their counterparts in the control groups. GSH-Px activity was significantly lower in the acute exercise group, and higher in the trained group than those as controls. SOD decreased in the elderly, smoking and acute exercise groups and increased in trained individuals. There was a significant increase in lipid peroxidation activity and a significant decrease in antioxidant enzyme activity in cases of acute exercise and smoking as well as the elderly.

Placental to fetal transfer of mercury and fetotoxicity.

Abstract: Mercury vapor is known penetrate the placental barrier more easily than inorganic mercury. A relative amount of mercury accumulates in the fetus after exposure of pregnant animals to mercury vapor. Mercury concentration in fetal organs is much lower than that in maternal organs except the liver, and fetal liver shows significantly higher mercury concentrations than maternal liver. In fetal liver, a substantial portion of mercury is bound to metallothionein (MT), which plays an important role as a reservoir of mercury during the prenatal period. The mercury retained in fetal liver is redistributed to other organs, such as the brain and kidney, with diminishing MT levels during postnatal development. Consequently, an increase in mercury concentration in the brain and kidney of the neonate is observed. In studies on animal offspring in utero exposed to mercury vapor, behavioral changes, such as radial arm maze, morris maze and lever-press durations, are observed when the levels of mercury vapor exceed the threshold limit value (TLV).

Lipid peroxidation and erythrocyte antioxidant enzymes in patients with Behçet's disease.

Abstract: In spite of unknown etiology, it is now accepted that reactive oxygen species (ROS) produced by neutrophils may be related to the pathogenesis of Behcet’s Disease (BD). The objective was to investigate whether increased production of ROS may affect erythrocyte oxidant/antioxidant system in patients with BD. The levels of malondialdehyde (MDA), one of the end products of lipid peroxidation, in plasma and erythrocyte, and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), antioxidant enzymes, in erythrocyte, also C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were measured in 22 patients in active stage of the disease and also in 30 healthy controls. Increased CRP, ESR, and MDA levels in plasma and erythrocyte and increased SOD but decreased GSH-Px activities in erythrocytes were observed in the patients, when compared to the controls. In addition, significantly positive correlations between plasma and erythrocyte MDA levels, and erythrocyte MDA-CRP, MDA-ESR, MDA-SOD, SOD-ESR and SOD-CRP levels, but negative correlation between plasma MDA and erythrocyte GSH-Px, were found in BD patients. In may be suggested that increased production of ROS in BD, as reflected by higher plasma and erythrocyte MDA levels, may impair erythrocyte membrane integrity and also may lead to the alterations in the erythrocyte antioxidant defense system, as reflected by higher SOD and lower GSH-Px activities in erythrocytes.

Complications of IgA nephropathy in a non-insulin-dependent diabetes model, the Akita mouse.

Abstract: The Akita mouse, which has a mutation (Cys96Tyr) in the insulin 2 gene (Ins2Akita), is a model for diabetes. The male Akita mouse has diabetes, while females develop mild diabetes. This study aimed to investigate renal complications in the Akita mouse model, which has been maintained in a heterozygous state Ins2Akita (+/−) with a C57BL/6 background (B6). Renal function (BUN, creatinine), serum IgA concentrations and histological changes in the kidneys were evaluated in diabetic and control mice in a B6 background. Five each of male and female mice per group (diabetes vs. control) were killed at 10, 20, 30 and 40 weeks of age. The influence of major histocompatibility antigens (MHC) on renal complications was tested using male diabetic mice in a C3H/He (C3H) background. When compared with controls, diabetic males, but not females, developed impaired renal function with elevation of serum IgA after 30 weeks of age. Diabetic glomerulosclerosis advanced with age in both sexes. Diffuse granular mesangial deposits of IgA were detected by immunofluorescence microscopy in diabetic males after 20 weeks. We tested whether the IgA-associated lesions were influenced by MHC using diabetic males in a C3H background. Similar degrees of diabetic glomerulosclerosis and glomerular IgA deposition were found in mice with C3H and B6 backgrounds. Akita mouse is a unique mouse model showing both mesangial sclerosis and IgA nephropathy.

Developmental Neurotoxicity Following Prenatal Exposures to Methylmercury and PCBs in Humans from Epidemiological Studies

Abstract: Adverse health effects following prenatal exposures to methylmercury (MeHg) have been apparent from several prospective cohort studies conducted in a fish-eating population. A prospective study in a Faroese birth cohort documented subtle deficits of several functional domains at prenatal MeHg exposure levels previously thought to be safe. Recent additional studies also showed neurobehavioral deficits associated with exposures to polychlorinated biphenyls (PCBs) with concomitant MeHg poisoning. In contrast, a prospective study in the Seychelles did not detect a similar association between MeHg exposure and neurodevelopmental deficits; children of the highest MeHg exposure group showed better scores in some developmental tests than those of the lower exposure groups for both prenatal and postnatal MeHg exposures. This paradoxical difference between both studies is summarized herein. The primary source of human exposure to MeHg is fish. Since a considerable number of pollutants, including polychlorinated biphenyls (PCBs) and pesticides, are also present in fish, and since some organochemical substances including PCBs are also well documented to be neurotoxic to the developing brain from epidemiological studies, the combined effects of these pollutants should be considered in discussing the neurotoxicity of MeHg. In this article, therefore, major prospective cohort studies focusing on the exposures to PCBs were reviewed.

Single nucleotide polymorphisms in the 3' untranslated region of vascular endothelial growth factor gene in Japanese population with or without renal cell carcinoma.

Abstract: We investigated native Japanese subjects whether C702T, C936T and G1612A polymorphisms in the 3' untranslated region (3'-UTR) of vascular endothelial growth factor (VEGF) gene are associated with the risk of renal cell carcinoma (RCC). Genomic DNAs from 145 RCC patients and 145 healthy controls were examined by polymerase chain reaction-based restriction fragment length polymorphism. Variant allele frequencies of C702T, C936T and G1612A were 0.00, 0.20 and 0.13 in the controls, respectively. The C702T and G1612A allele frequencies were significantly different between the Japanese population and the Caucasian population reported elsewhere. For each of C936T and G1612A polymorphisms, there was no statistically significant difference in the distribution of genotype frequencies between the cases and controls. Odds ratios and 95% confidence intervals computed by logistic regression analyses were not statistically significant. Stratification for the RCC cases according to pathological cell subtype, grade or stage failed to reveal any significant heterogeneity with respect to the genotype of each VEGF polymorphism. We revealed that there are significant ethnic differences in the C702T and G1612A allele frequencies, but suggested that C702T, C936T and G1612A polymorphisms in the 3'-UTR of VEGF gene are not associated with the risk of RCC, at least in Japanese population.

N-Acetylcysteine Reduced the Effect of Ethanol on Antioxidant System in Rat Plasma and Brain Tissue.

Abstract: Chronic ethanol administration is able to induce an oxidative stress in the central nervous system. N-Acetylcysteine (NAC) has antioxidant properties; as a sulphydryl donor, it contributes to the regeneration of glutathione and it acts through a direct reaction with hydroxyl radicals. In this study we investigated a possible beneficial effect of NAC on some of the free radical related parameters. Twenty four male Wistar rats were divided in to three groups and were given ethanol (Group 1), ethanol and NAC (Group 2) and isocaloric dextrose (Group 3). Ethanol and NAC were given intragastrically at doses of 6 g/kg/day and 1 g/kg/day, respectively. Our results show that chronic ethanol intake elicits statistically significant increase in MDA and NO levels and decrease in SOD and GSH levels in both plasma and brain (p < 0.001). GPx levels decreased in erythrocytes (p < 0.001). CAT activity showed significant decrease only in brain samples (p < 0.001). NAC administration effectively restores the above results to nearly normal levels. Therefore we suggest that reactive free radicals are, at least partly, involved in the ethanol-induced injury of brain cells and NAC mitigate the toxic effects of ethanol on the oxidant-antioxidant system of rat plasma and brain.

Effect of recombinant basic fibroblast growth factor (bFGF) on fibroblast-like cells from human rotator cuff tendon.

Abstract: Rotator cuff tendon cells (RCC) derived from surgical samples showed fibroblast-like morphology. Histological staining demonstrated collagen secretion by RCC. Immunohistological findings revealed that RCC secreted type I and III collagen, but not type II collagen. In addition, the SDS-PAGE analysis suggested that RCC predominantly produced type I collagen. Basic fibroblast growth factor (bFGF) had a stimulatory effect on the proliferation of RCC dose-dependently up to 1 ng/ml. Administration of bFGF suppressed the secretion of collagens from RCC in a dose-dependent manner.

Autonomic nervous functions in fetal type Minamata disease patients: assessment of heart rate variability.

Abstract: In order to assess the cardiovascular autonomic nervous functions in patients with fetal type Minamata disease (FMD), we investigated blood pressure (BP), and conducted time and frequency domain analysis of heart rate variability (HRV). Subjects were 9 patients in Meisuien recognized as FMD, and 13 healthy age matched control subjects. HRV and BP were assessed after subjects rested in a supine position for 10 minutes. Electrocardiographic (ECG) data were collected for 3 minutes during natural breathing. Time domain analysis (the average of R-R intervals [Mean RR], standard deviation of R-R intervals [SD RR], coefficient of variation [CV]), and frequency domain analysis by fast Fourier transformation (FFT) (power of low frequency [LF] and high frequency [HF] component, expressed in normalized units [nu]) were then conducted. In the time domain analysis, the mean RR of the FMD group was significantly lower than that of the control group. Neither SD RR nor CV showed significant differences between the two groups, but both tended to be lower in the FMD group. In the frequency domain analysis, the HF component of the FMD group was significantly lower than that of the control group. Pulse pressure (PP) was significantly lower in the FMD subjects. These findings suggest that parasympathetic nervous dysfunction might exist in FMD patients, who were exposed to high doses of methylmercury (MeHg) during the prenatal period. Decrease of PP might be due to degenerative changes of blood vessels driven by exposure to high doses of MeHg.

Cellular cadmium uptake mediated by the transport system for manganese.

Abstract: The mechanism of cellular cadmium (Cd) uptake has been poorly understood Recently, we developed Cd-resistant cell lines from metallothionein null mouse cells and showed that the Cd resistance of these cells was conferred primarily by a reduced Cd accumulation Surprisingly, the uptake rate of manganese (Mn) was also markedly reduced in Cd-resistant cells Subsequent studies on the kinetics of Cd and Mn uptake by Cd-resistant and parental cells revealed that the Mn transport system with high affinity for Mn is used for cellular Cd uptake, and that this pathway is suppressed in Cd-resistant metallothionein null cells This is the first indication that the transport system for Mn is used for Cd uptake in mammalian cells Divalent metal transporter 1 (DMT1) is the only known mammalian transporter involved in the uptake of both Cd and Mn However, the high-affinity Mn/Cd transport system we found seems to be distinct from DMT1 because of the difference in optimal pH and substrate specificity On the other hand, various types of Mn transporters have been shown to play an important role in cellular Cd uptake in non-mammalian species such as yeast, plants and bacteria, suggesting the existence of Mn transporters other than DMT1 in mammals

Medial superior cluneal nerve entrapment neuropathy in teenagers: a report of two cases.

Abstract: Medial superior cluneal nerve entrapment neuropathy causes pain radiating from the low back down to the posterior thigh. It tends to be misdiagnosed as a lumbar spine disorder. Patients in previous reports were in the middle or old age at the onset. Proposing simultaneous full flexion of the ipsilateral hip and knee joints as a provocation test, we present two cases of teenager females who spent long before diagnosis of their condition. Both of them had engaged in vigorous sports activities and completely recovered from the disability following local anaesthetic and corticosteroid injection at the trigger point.

Effects of cadmium intake on bone metabolism of mothers during pregnancy and lactation.

Abstract: Cadmium (Cd) is a heavy metal that exists ubiquitously in the environment, and it interacts with essential elements such as zinc, copper, iron, and calcium (Ca). Particularly, Cd interferes with Ca and vitamin D metabolism in bone kidney and intestine. The interaction between Cd and Ca in bone, intestine, and kidney may result in the disorder of bone metabolism. On the other hand, pregnancy and lactation are also important physiological factors affecting bone metabolism in the mother. Ca absorption is decreased by competition with Cd in the intestine, and more Ca is released from maternal bone and transferred to neonate by lactation. In the intestine, Cd uptake competes with Ca uptake. Cd causes a marked decrease in bone density compared to the normal decrease in bone mineral density during lactation. Lactation is an important factor contributing to the decrease in bone mineral density and Cd has an additive effect of decreasing bone metabolism of mother animal, although the Cd intake level is relatively low (approximately 3-14 microgCd/kg/day). The relationship among maternal Cd intake, renal function and bone metabolism and the interaction between Cd and Ca during lactation are reviewed herein, together with additional data obtained recently in our laboratory.

Lead, Chemical Porphyria, and Heme as a Biological Mediator

Abstract: One of the most well-characterized symptoms of lead poisoning is porphyria. The biochemical signs of lead intoxication related to porphyria are δ-aminolevulinic aciduria, coproporphyrinuria, and accumulation of free and zinc protoporphyrin in erythrocytes. From the 1970s to the early 80s, almost all of the enzymes in the heme pathway had been purified and characterized, and it was demonstrated that δ-aminolevulinic aciduria is due to inhibition of δ-aminolevulinate dehydratase by lead. Lead also inhibits purified ferrochelatase; however, the magnitude of inhibition was essentially nil even under pathological conditions. Further study proved the disturbance of iron-reducing activity by moderate lead exposure. Far different from these two enzymes, lead failed to inhibit purified coproporphyrinogen oxidase, i.e., the mechanism of coproporphyrinuria has not yet been understood. During the 80s to the 90s, the effects of environmental hazards including lead were elucidated through stress proteins, indicating the induction of some heme pathway enzymes as stress proteins. At that time, gene environment interaction was another focus of toxicology, since gene carriers of porphyrias are considered to be a high-risk group to chemical pollutants. Toxicological studies from the 70s to the 90s focused on the direct effect of hazards on biological molecules, such as the heme pathway enzymes, and many environmental pollutants were proved to affect cytosolic heme. Recently, we demonstrated the mechanism of the heme-controlled transcription system, which suggests that the indirect effects of environmental hazards are also important for elucidating toxicity, i.e., the hazards can affect cell functions through such biological mediators as regulatory heme. It is, therefore, probable that toxicology in the future will focus on biological systems such as gene regulation and signal transduction systems.

Odor perception in patients with multiple chemical sensitivity.

Abstract: Since symptoms typical for multiple chemical sensitivity (MCS) are induced by exposure to low levels of chemicals, we hypothesize that MCS represents an impaired recognition of odors or an increased emotional reaction to common odors. Twenty-five subjects with MCS, 20 women and 5 men, and 50 gender-and-age matched controls participated in this study. The University of Pennsylvania Smell Identification Test (UPSIT) and the Cross-Cultural Smell Identification Test (CC-SIT) were administered. In addition to selecting the most probable odor among the four, the subjects were asked their impression of each odor. Odor identifiability evaluated by the scores of two tests, were almost equal in MCS and control groups. The mean CC-SIT odor per person with pleasant feeling was lower in MCS than in controls. The mean odor per person creating an unpleasant sensation was higher in MCS than in the controls. Gingerbread was the only odor making MCS subjects more pleasant than the controls. Nine out of 40 UPSIT odors were felt as unpleasant by MCS subjects more than by controls. This study indicates that MCS subjects are able to identify the odors equally as well as the controls but feel unpleasant to a larger number of odors than the controls. Despite unknown mechanisms of the altered odor perception in MCS, the application of these tests for diagnostic procedure of MCS is proposed.

Relationship of health status and social support to the life satisfaction of older adults

Abstract: We examined the relationship of health factors and social support to life satisfaction in older adults dwelling in a rural town. The gender difference in variables related to life satisfaction was also discussed in this study. One hundred and forty-two older adults (86 females and 56 males) who completed a self-administered questionnaire and participated in a health examination in 1998 or 1999 comprised the study participants. The t-test and chi-square test were used to assess the differences between the two genders. Correlation measure and multiple regression analysis were used to assess the relationship between life satisfaction and other health related or socially related factors for each gender. Significant gender differences were observed in living status and several health related factors. According to the results of the multiple regression analyses, life satisfaction was related to mental health and age in females, while it was related to mental health status and social support from others in males. Gender differences in the variables associated with life satisfaction were observed among the community-dwelling older adults. These data suggest the importance of mental health for older adults. When preparing health promotion strategies for older adults, results of gender differences as they related to social support and life satisfaction should be applied in practice.

The effect of vitamin E treatment on oxidative stress generated in trained rats.

Abstract: The aim of this study was to investigate the effect of vitamin E treatment on increased oxidative stress in rats exposed to a swimming exercise protocol. In order to examine the effects of physical swimming training on the antioxidant defences of tissues and on their susceptibility to damage induced by exercise, the levels of glutathione (GSH) and thiobarbituric acid reacting substances (TBARS) levels, on indicator of lipid peroxidation in various tissues, have been determined. In this study, four groups of female rats were used while the rats were trained to swim for 30 minutes a day and five days a week which lasted eight weeks and vitamin E (vit. E) supplementation (30 mg/kg/day) has been carried out for five days a week. TBARS levels are significantly found lower in both trained and sedentary vit. E supplemented groups, since vit. E is the most important antioxidant in an earlier line of defence in lipid peroxidation. Also, in vit. E supplemented trained rats, the glutathione response is observed to be significantly higher, supporting with the TBARS levels and in accordance with the literature. But in the sedentary group without vit. E supplementation, the GSH levels of the liver and the heart tissues were significantly lower than both vit. E supplemented sedentary and trained groups. These results evaluate that vit. E confers protection to GSH levels in these tissues where the GSH levels were found significantly lower in the groups not supplemented with vit. E.

Ultrastructural Localization of Aquaporin 4 and α1-Syntrophin in the Vascular Feet of Brain Astrocytes

Abstract: Aquaporin 4 (AQP4) is a recently discovered membrane bound water-selective channel and has been described at the light microscopic level to be predominantly expressed in the astrocytes of the brain, especially at the perivascular astrocyte endfoot processes. Alpha1-syntrophin, a member of dystrophin-associated protein, has also been reported at the light microscopic to be expressed level in the same site of astrocytes as AQP4 and interacts with other molecules through its PDZ domain. AQP4 expression has been reported to be absent at the sarcolemma and the perivascular astrocyte endfoot processes of alpha1-syntrophin knockout mice. Based on these observations, the molecular association between AQP4 and alpha1-syntrophin could be speculated. To test this hypothesis, we investigated the ultrasturctural localization of AQP4 and alpha1-syntrophin in the brain astrocytes by using double immunogold labeled electron microscopy. The results showed that AQP4 and alpha1-syntrophin colocalized frequently at the astrocyte membrane, especially at the perivascular astrocyte endfoot processes and suggested the presence of linkage between AQP4 and alpha1-syntrophin at the astrocyte plasma membrane.

Investigation of intracellular factors involved in methylmercury toxicity.

Abstract: Methylmercury is a known pollutant that causes severe central nervous system disorders. It is capable of passing through the blood-brain barrier and accumulates in cerebral cells. However, little is known regarding the mechanism of its toxicity at the molecular level. Using yeast cells, we searched for the genes involved in the expression of methylmercury toxicity, and found that genes encoding L-glutamine·D-fractose-6-phosphate amidotransferase (GFAT) and ubiquitin transferase (Ubc3) confer methylmercury resistance on the cells. It has also been shown that GFAT is the target molecule of methylmercury in yeast cells. These findings provide important clues about the mechanism underlying methylmercury toxicity ini mammals.

Clinical roles of serum autoantibody against neuron-specific enolase in glaucoma patients.

Abstract: In our recent papers, we found the presence of serum autoantibody against neuron specific enolase (NSE) in some glaucoma patients and suggested that this antibody might have significant roles in pathogenesis of glaucomatous optic neuropathy. In order to evaluate further the clinical roles of serum autoantibody against NSE in glaucoma, serum autoantibody against NSE was examined by western blot analysis and enzyme-linked immunosorbent assay (ELISA) in 4 patients with ocular hypertension (OH) and 242 patients with glaucoma (normal tension glaucoma [NTG], 73 cases; primary open angle glaucoma [POAG], 169 cases), and the relationships between the titers of anti-NSE antibody and clinical characteristics were evaluated. The titers of anti-NSE antibody showed a regular decreasing pattern with deteriorating visual field losses and glaucoma stages in POAG, especially early and late stages. However, no systematic pattern was observed in NTG. Although maximum and mean intraocular pressures (IOP)s and progression of visual field losses showed no correlation with the levels of serum anti-NSE antibody titers in either POAG or NTG, the anti-NSE antibody titers were relatively higher in NTG with visual field deterioration than in those without it. The present observations suggest that serum autoantibody against NSE may be clinically useful for diagnosing early stages of POAG, and for monitoring glaucoma progression of NTG.

Effects of Octreotide Acetate and Saccharomyces Boulardii on Bacterial Translocation in an Experimental Intestinal Loop Obstruction Model of Rats

Abstract: Intestinal obstruction (IO) induces bacterial translocation (BT) due to mucosal disruption, motility dysfunction, and increased intestinal volume, leading to bacterial overgrowth. This study was conducted to investigate the effects of octreotide acetate (OA) and Saccharomyces boulardii (SB) on the BT and intestinal integrity in an animal model of intestinal loop obstruction (LO). Forty adult male Sprague-Dawley rats (250-300 g) were randomized into 4 groups containing 10 rats each. Complete IO was created in the distal ileum of rats by a single 3-0 silk suture (LO). Group Sham: Sham (Laparotomy only was performed in this group); group LO: LO; group OA: LO plus OA (100 μg/kg, at 0, 12 hours of obstruction); group (SB): LO plus SB (800 mg/kg/day, via orogastric and preoperative for 3 days). After 24 hours, samples of mesenteric lymph nodes (MLN), liver, spleen and blood were obtained and cultured. The terminal ileum specimens were examined histopathologically. There were no BT in group Sham, but BT was noticed totally in 31 (77.5%) cultures in group LO. This rate was reduced to 30% (n=12), 10% (n=4) in the groups OA and SB respectively. Bacterial translocations of MLN and the liver in group LO were significantly higher than those of groups OA and SB. Bacterial translocations of the both spleen and blood in group LO were significantly higher than those of groups OA and SB. The mean bacterial counts, colony-forming units per gram tissue (cfu/g), in the MLN, liver and spleen of group LO were found significantly higher than those of groups OA and SB. The mean villus height in group OA was significantly higher than that of group LO and it in the group SB significantly higher than those of groups LO and OA. The present experimental study has demonstrated that OA and SB may have protective effects against BT in mechanical bowel obstruction and additionally SB preserves intestinal mucosal integrity.

Clinical value of color Doppler ultrasonography measurements of full-term newborns with perinatal asphyxia and hypoxic ischemic encephalopathy in the first 12 hours of life and long-term prognosis.

Abstract: The cerebral blood flow velocities (CBFV) of infants with perinatal asphyxia and hypoxic ischemic encephalopathy (HIE) in the first 12 hours of their lives have been the chief focus of our concern in this study. Cerebral ischemia which can develop in the earlier hours of HIE, and the detection and diagnosis of this condition with color Doppler ultrasonography (cD-USG) will be put into discussion. Twenty-three full-term newborn infants who had perinatal asphyxia and HIE together with a control group constituting twenty full-term newborn infants who produced no problems, were included in our study. All of the infants underwent cD-USG in the postpartum period of the first 12 hours (mean 8.4 hours). Measurements being based upon peak systolic velocity (PSV), end diastolic velocity (EDV) and Pouecelout's resistive index (RI) in anterior and middle cerebral arteries were conducted. The infants, having been discharged from the unit they were followed up for mean 9.8 months in the outpatient clinic. PSV and EDV counts in the postpartum first 12 hours of 23 infants who were detected to have HIE were found to be significantly lower compared to the control group, whereas RI counts were found to be significantly higher (p < 0.05). The counts obtained from the right and left cerebral arteries of the infants with HIE were found to be corraleted with each others. The neonates in the patient group were observed to have gone through this prognosis: Three of them died, three of them had cerebral palsy, one of them had infantile spasms, and three of them had developmental retardation. When we compared the CBFV of the 10 neonates who had poor prognosis, retrospectively with the other 13 neonates who had good prognosis, PSV and EDV were found to be significantly lower and RI significantly higher (p < 0.05). In the light of the data we have obtained, cD-USG can be considered to be a highly practical device in evaluating CBVF of the infants with HIE. A skillful detection of the decrease in cerebral blood flow which can develop in the postasphyxial first 12 hours and the prospective treatments being based upon this approach would contribute to the diagnosis, treatment and prognosis of such cases.

Comparative immunohistochemical localizations of aquaporin-1 and aquaporin-4 in the cochleae of three different species of rodents.

Abstract: The species-specific difference of the immunohistochemical localization of aquaporin-1 (AQP1) and aquaporin-4 (AQP4) was investigated in the cochleae of the 3 different species of rodents, including guinea pig, mouse and Mongolian gerbil. In the guinea pig cochlea, intense AQP1-like immunoreactivity was present in the type III fibrocytes in the spiral ligament and the mesenchymal cells just below the basilar membrane. Immunostaining was also found in some type IV fibrocytes in the spiral ligament, fibrocytes in the spiral limbus and mesenchymal cells lining the perilymphatic space against the bony otic capsule. In contrast, no remarkable immunostaining was found in the basilar membrane of the mouse cochlea. The medial part of the Reissner's membrane was positively immunostained with anti-AQP1 antibody only in the mouse cochlea. In the gerbil cochlea, AQP1-like immunoreactivity was weak compared with the other 2 species. AQP4 was found in the cochlear supporting cells, including Claudius cells, Hensen's cells and inner sulcus cells of the 3 rodent species. AQP4 was also expressed in some interdental cells of the spiral limbus. Weak immunoreactivity was also found in the root cells only in the upper turns of the guinea pig cochlea. In contrast, no detectable immunoreactivity was found in the root cells of the other 2 species. The results obtained in the present study provide the first evidence for the existence of the species differences in the expression of the AQP1 and AQP4 proteins in the rodent cochlea.

Paradigm Shift in Zinc: Metal Pathology

Abstract: Zinc (Zn) is an essential, common metal in animal tissues. Zn levels were elevated in only four tissues after Zn administration, the highest increase being in the pancreas. Zn concentration was increased by metallothionein induction. Metallothionein-bound Zn significantly reduced the toxicity of the metals Cd, Cu and Hg. It should be noted that tissue Zn levels are different in experimental animals and humans. Acute pancreatitis was observed following the injection of a large dose of Zn. Different metals have different target organs. Using metal pathology, treatments may be developed to save patients suffering from hepatic and renal diseases because Zn is used to a model animal of hepatic or renal disease.

Congenital central hypoventilation syndrome: a novel mutation of the RET gene in an isolated case.

Abstract: Recently, a few genetic abnormalities were identified in congenital central hypoventilation syndrome (CCHS or Ondine's curse). CCHS is often associated with other neurocristopathies, especially with Hirschsprung's disease (HSCR). Mutations of the genes involved in the receptor tyrosine kinase RET (REarranged during Transfection) (RET)-glial cell line-derived neurotrophic factor (GDNF) and/or endothelin 3 (EDN3)-endothelin receptor-B (EDNRB) signaling pathway have been found in some of HSCR patients. In this study, we analyzed candidates for HSCR, namely the RET, GDNF, EDN3 and EDNRB genes in three isolated CCHS patients to confirm the hypothesis that some CCHS patients have a common genetic abnormality with patients having HSCR or other neurocristopathies. We found a novel R114H mutation of the RET gene in one patient. The R114H mutation is unlikely to be a polymorphism and appears to be associated with CCHS. In addition, we also examined the HOX11L2 (RNX) gene, for which knock-out mice showed CCHS-like syndrome in these isolated CCHS patients and did not detected any mutation. Further cases should be analyzed for more candidates to clarify the pathophysiology of CCHS.

The effect of intracerebroventricular dopamine administration on the respiratory response to hypoxia.

Abstract: Acute hypoxia produces an increase in ventilation. When the hypoxia is sustained, the initial increase in ventilation is followed a decrease in ventilation. The precise mechanism of this decline in ventilation during sustained hypoxia is unknown. Recent studies hypothesized that the accumulation of dopamine in the central nervous system might have a major role in production of hypoxic respiratory depression. The purpose of this study was to examine whether dopamine has an effect on occurance of central ventilatory depression which is seen in acute hypoxia in peripheral chemoreceptors denervated animals. The experiment were conducted in rabbits anesthetized with Na-pentobarbital (25 mg·kg-1 i.v.). For intracerebroventricular (ICV) injections of dopamine (1 μg) in each animal, canula was placed in left lateral cerebral ventricle by stereotaxic method. Respiratory frequency (f·min-1), tidal volume (VT) ventilation minute volume (VE) and systemic arterial blood pressure (BP) were recorded during air and 3 minutes hypoxic gas mixture (8%O2-92%N2) breathing. ICV administration of dopamine during normoxia decreased VT, f, VE and BP, significantly. When rabbits were injected with an ICV dopamine on hypoxic gas mixture breathing in control animals, there was depression of hypoxic ventilatory responses. After ICV administration of dopamine antagonists haloperidol (0.1 mg) and domperidone (0.07 mg) in chemodenervated rabbits, the significant increases in VT, VE and BP were observed. On the breathing of hypoxic gas mixture of chemodenervated and ICV dopamine antagonists administrated rabbits, hypoxic depression was completely abolished. These results of this study show that accumulation of dopamine in the brain seems to reduce the response of the central control mechanisms to chemoreceptor impulses during normoxia and hypoxia. In conclusion present study suggests important role played by central dopaminergic pathways in the occurence of acute hypoxic ventilatory depression.

Metacarpal Bone Mineral Density, Body Mass Index and Lifestyle among Postmenopausal Japanese Women : Relationship of Body Mass Index, Physical Activity, Calcium Intake, Alcohol and Smoking to Bone Mineral Density : The Hizen-Oshima Study

Abstract: The present study was designed to investigate the influence of modifiable risk factors (body weight and lifestyle) for bone loss on bone mineral density (BMD). We examined age-specific changes in metacarpal BMD, and its associations with body mass index and lifestyle among 532 community-dwelling postmenopausal Japanese women. Measurements of the second metacarpal BMD were obtained from the hand radiographs using computer-assisted radiographic absorptiometry. Body height and weight were measured, and body mass index (BMI) was calculated. Physical activity index was calculated using validated questionnaire. Daily calcium intake and amount of ingested alcohol were estimated by semiquantitative food frequency questionnaire. Current smoking status was obtained by questionnaire. Metacarpal BMD decreased significantly with increasing age. Simple correlation analysis indicated that metacarpal BMD correlated significantly with BMI and physical activity index. On the other hand, metacarpal BMD did not correlate with calcium intake and alcohol drinking. Metacarpal BMD in current smokers was not different from that in nonsmokers. Multiple regression analysis showed that increasing age was associated with decreased metacarpal BMD and greater BMI increased metacarpal BMD. However, physical activity, calcium intake, alcohol drinking and current smoking were not significant determinants of metacarpal BMD. Our findings suggest that maintenance of adequate body mass (prevention for leanness) is important for prevention of postmenopausal bone loss.

Establishment and Characterization of a Simian Virus 40-Immortalized Rat Pancreatic Stellate Cell Line

Abstract: Activated pancreatic stellate cells (PSCs) have recently been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line would be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish an immortalized cell line of rat PSCs. PSCs were isolated from the pancreas of male Wistar rats, and the simian virus 40 T antigen was introduced to PSCs by retrovirus-mediated gene transfer. This procedure yielded an actively growing cell line, designated as SAM-K. This cell line has been passaged repeatedly for almost 2 years, and is thus likely immortalized. SAM-K cells retained morphological characteristics of primary PSCs, and expressed α-smooth muscle actin, glial fibrillary acidic protein, type I collagen, fibronectin, and prolyl hydroxylases. The level of p53 expression was very high in SAM-K cells. Proliferation of SAM-K cells was stimulated by serum and platelet-derived growth factor-BB. Interleukin-1β (IL-1β) activated nuclear factor-κB, activator protein-1, and three classes of mitogen-activated protein (MAP) kinases: extracellular signal-regulated kinase1/2, c-Jun N-terminal kinase, and p38 MAP kinase. IL-1β induced expression of intercellular adhesion molecule-1 and monocyte chemoattractant protein-1, both of which were abolished in the presence of pyrrolidine dithiocarbamate, a specific inhibitor of nuclear factor-κB activation. IL-1β-induced monocyte chemoattractant protein-1 was partially inhibited by specific inhibitors of MAP kinase kinase (U0126) and of p38 MAP kinase (SB203580) whereas intercellular adhesion molecule-1 expression was not altered by the inhibitors. Thus, SAM-K would be useful for in vitro studies of cell biology and signal transduction of PSCs.