A total number of 668 apparently healthy fish were obtained from farm to study the effect of two heavy metals (Copper and Mercury) on histopathology of liver, kidney, spleen, gills and muscles also residues in muscles. The $LC_{50}$/96 hr. of Cu and Hg were estimated and fish exposed to 1/2 $LC_{50}$ for 7 days and for 1/10 $LC_{50}$ for 8 weeks from each product separately. Histopathological findings in acute and chronic mercuric chloride toxicity revealed degeneration and necrosis in the glomeruli, interstitium tissue and epithelium lining renal tubules. The tubular epithelium became necrotic at several places. Eosinophilic hyaline droplets is exist in the cytoplasm of the necrosed cells. Degenerative changes and hyperactivity in melanomachrophage center was seen in the spleen together with some necrotic areas. Necrosis and aggregation of melanomachrophage were seen in the hepatic cells, Hepatic cells showed vacuolar degeneration in the hepatic cells. Gills showed loss in the lamellae of the filaments associated with edema, inflammatory cells infiltration and haemorrhages in the arch. The sarcoplasm of the bundles of the skeletal muscle showed granular degeneration and focal inflammatory cells infiltration between the hyalinized bundles. Mercury residues obtained from these studies in the acute toxicity were 0.22 ppm/gm in the 2nd day, 0.411 ppm/gm in the $5^{th}$ day ended with 0.96 ppm/gm in the $7^{th}$ day. In chronic toxicity it was 1.1320, 1.7140, 2.3620 and 3.5640 ppm/gm respectively from the $2^{nd}$ to the $8^{th}$ week of exposure. In acute and chronic copper toxicity, there was degenerative changes in renal tubules. Melanophores aggregation in the wall of the blood vessels of the spleen and depletion of some of the melanophores in the melanomachrophage were seen together with necrosis in some areas. Congested Mvs (Micro vessels) and vacuolation of hepatocytes were observed. Some areas of hemorrhage and melanophores vacuolar degeneration in the liver were seen. There was mitosis in some areas with displesia of hepatopancreatic cells and eosinophilic granular cells aggregation. Zymogen granules disappeared and there were dyplastic hepatocytes. Congestion in the blood vessels of the gill filaments, associated with massive number of granular eosinophilic cells infiltration were seen in the base of the filaments. There were sever vacuolization and hyalinization in the skeletal muscle bundles. Detection of residues of copper sulfate revealed increase of the amount of copper measured in ppm/gm comparing to the normal control starting from 0.60 ppm/g in the $2^{nd}$ day, 0.67 ppm/g in the $5^{th}$ day and 0.67 ppm/g in the $7^{th}$ day. Result obtained in chronic copper sulfate toxicity revealed gradual increase of the amount of copper which ranged from 0.18 ppm/g at the $2^{nd}$ week to 0.21 ppm/g in the $8^{th}$ week of exposure.
Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.
Objective : The aim of the study was to investigate the preventive effect of Acanthopanax senticosus(AS) aqua-acupuncture into Sinsu(BL23) of the multiple low-does strepozotocin(STZ)-induced diabetic rats. Methods : The experimental animals were divided into 4 groups : normal group of rats, control group of multiple low-does STZ-induced diabetic rats, NSAA group with 0.4ml normal saline(NS) aqua-acupunctured subcutaneously into Sinsu in multiple low-does STZ-induced diabetic rats, and ASAA group with 0.4ml of 20% AS aqua-acupunctured subcutaneously into Sinsu in multiple low-does STZ-induced diabetic rats. Each of AS and NS aqua-acupuncture was done subcutaneously into both loci of Sinsu taking turns everyday for 3 weeks. Thereafter the levels of serum glucose, body weight, index of kidney hypertrophy, urine glucose, urinary albumin excretion, creatinine clearance, mesangial cell and TGF-${\beta}1$ expression in glomeruli and tubular cells were measured on the determined day. Conclusions : 1. Both ASAA and NSAA groups decreased the serum glucose levels in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group showed more significant decreases than NSAA group. 2. Both ASAA and NSAA groups prevented the development of diabetes in multiple low-dose STZ-induced diabetic rats as compared to the control group, and ASAA group prevented more markedly the development of diabetes than NSAA group. 3. Both ASAA and NSAA groups prevented the reduction of body weight in multiple low-dose STZ-induced diabetic rats as compared to the control group, and ASAA group showed the same as the normal group. 4. Both ASAA and NSAA groups did not show any changes of the creatinine clearance in multiple low-does STZ-induced diabetic rats. 5. Both ASAA and NSAA groups prevented the excretion of urinary glucose and albumin in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group showed more significant prevention than NSAA group. 6. Both ASAA and NSAA groups prevented the expansion of glomerular cells and the protein expression of transforming growth factor-${\beta}1$ in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group prevented more significantly than NSAA group.
We investigated the effects of vitamin E supplementation on protein glycosylation in early and end stage product, and light microscopic studies were done on the renal glomeruli of KK-mice of various ages and various duration of diabetes. Weaned KK-mice were fed high fat diets containing 20% corn oil(wt/wt), and sacrificed at 4,6, and 9 months of age. The high vitamin E diet was a high fit diet supplemented with an excess amount of d1-$\alpha$-tocopheryl acetate (2080IU/kg diet). We measured Hemoglobin $A_{IC}$ (Hb $A_{IC}$) as a glycosylation early product, and renal collagen-linked fluorescence as a glycosylation end product. In the diabetic group, levels of Hb $A_{IC}$ were increased within 2 months after onset of diabetes and remained at a constant level for the duration of experiment. 5 months after onset of diabetes, renal collagen linked fluorescence(CLF) was markedly increased. A quantative, morphologically demonstratable, progressive thickening of the basement membrane and calcification occured in the diabetic KK-mice. There is a statiscally positive correlation between CLF and histologic grade of diabetic nephropathy. Hepatic vitamin E levels correlated with those of Hb $A_{IC}$, renal CLF, and renal calcification. Treatment with vitamin I did not modify the level of blood glucose. However, we observered a significant lowering of CLF and Hb $A_{IC}$ in diabetic mice. Supplementation of vitamin E was found to delay the progression of diabetic nephropathy. (forean J Nutrition 31(6) : 1024-1030, 1998)0, 1998)
For the purpose of stydying the pharmacodynamic action of methemoglobin, the author made the following experiments: 1. Preparation of hemoglobin and methemoglobin solutions: Red cell suspension from rabbit blood was hemolysed with distilled water and then divided into two portions. One portion was dialysed through cellophane paper and made isotonic with the proper amount of sodium chloride. The second portion was treated with sodium nitrite to convert hemoglobin to methemoglobin, dialysed through cellophane paper and made isotonic. 2. The concentration of methemoglobin in solution, plasma and urine was determined by Horecker and Brackette's method, and that of hemoglobin by the cyanmethemoglobin method. 3. The concentration of methemoglobin and hemoglobin in the plasma and urine of rabbits was measured at several intervals of time after infusion of the above samples. 4. The blood pressure and respiration of rabbits were recorded on a kymograph, and the effects of the samples on them were observed. 5. The effects of the samples on the movements of the in-situ heart and the isolated intestine of rabbits were studied. 6. The kidneys of rabbits were excised 4 to 5 hours after injection of the samples, and histopathological examinations were made. These experiments revealed the following results: 1. When methemoglobin solution was allowed to stand in room air, there was no decrease in the concentration of methemoglobin. 2. When methemoglobin solution was mixed with whole blood and incubated at $37^{\circ}C$, the concentration of methemoglobin decteased gradually. 3. After the infusion of methemoglobin and hemoglobin solutions, the rate of disappearance of methemoglobin in the plasma was more rapid than that of hemoglobin in the plasma. The higher the initial concentration in the plasma, the larger was the rate of disappearance of methemoglobin. 4. The rate of disappearance of methemoglobin was exceedingly rapid for 30 minutes after the infusion. 5. The urinary excretion of methemoglobin was more rapid than that of hemoglobin. 6. It would seem that the circulating blood contains substances which are promptly mobilized in the plasma to reduce methemoglobin to hemoglobin. 7. Moderate amounts of methemoglobin solution caused some rise in the blood pressure and a transient acceleration of the respiration of the rabbits. These effects of methemoglobin were milder than those of hemoglobin. 8. The movements of the in-situ heart and the isolated intestine of rabbits were accelerated by methemoglobin. These accelerating effects were milder than those of hemoglobin. 9. In the kidneys of rabbits treated with methemoglobin solution, hyperemia of the glomeruli, cloudy swelling and hemoglobin deposit in the tubular epithelium, hemoglobin casts in the tubular lumina of the proximal tubules, and interstitial congestion were constantly observed. There was no definite difference between the histological findings in the rabbit kidneys injected with methemoglobin, and those injected with hemoglobin solutions.
Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).
Mongolian gerbil (Meriones unguiculatus) has been as an animal model for studing the neurological diseases such as stroke and epilepsy because of the congenital incompleteries in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animals. In order to accomplish the this research, first of all another divided the laboratory animals 5 groups of which each group include the 5 animals. In this study were investigated the histological structure in the kidney, measured the plasma osmolalities at the time of sacrifice of indivisual animals, and the body weights every day during water-deprived. The results obtained in this study were summarized as followings: 1. The body weights and decreasing rates of the body weight in water-deprived mongolian gerbil groups were continuosly decreased. 2. The plasma osmolalities were increased from the 5th water-deprived day, after then the gradually increasing reached nearly its equilibrium state at the 10th water-deprived day. 3. The urine volumes were abruptly decreased from the 2th water-deprived day, after then the gradually decreasing patterns were reached nearly its equilibrium state at the 10th day, and stopped the 11th day. 4. In the light microscopical observation of the kidney, glomerular capillary loop thickening, mesangial matrix increasing, sclerosis, glomerular cystic atrophy, interstitial fibrosis, tubular dilatation, mononuclear interstitial inflammation, interstitial mineralization, and hyperplasia of the collecting duct epithelium in the cortex area, were observed from the 10th water deprived day, and the lesions were gradually severe changed as the time lapse. 5. In the electron microscopical findings of the kidney, the degenerative changes of endothelial cell, podocyte and mesangial cell in glomeruli were initially observed on the 10th water-deprived day as well as the degeneration of microvilli and intracellular organelle in the renal tubules.
Morphological difference of the renal glomerulus at different age groups have been studies in one week-old, five weeks-old, eight weeks-old, six months-old, twelve months-old, eighteen months-old, twenty-four months-old, and thirty months-old ICR mice. Pieces of the tissue taken from the renal corticies were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde solution (0.1 Millonig's phosphate buffer pH 7.3), and 1% osmium tetroxide solution (0.1 M Millonig's phosphate buffer, pH 7.3), and were embedded in araldite mixture. The ultrathin sections were stained with uranyl acetate and lead citrate solution, and were observed under a JEM 100CX-II electron microscope. The results were as follows: 1 In the one week-old mouse, thicknesses of the three layers of the glomeluar basal lamina (lamina densa, lamina rara interna and lamina rara externa) are similar, but in the five weeks-old mouse, thick lamina densa becomes a greater portion of the thickness of whole glomerular basal lamina. 2. No difference was noticed between thickness of the renal glomerular basal lamina of the five weeks-old mouse compare with that of the one week-old one, but basal lamina of the eight weeks-old one is thickened considerably and thicknesses were maintained through twelve months-old one. After eighteen months, the thickness of the glomerular basal lamina is increased remarkably. 3. After eighteen months, electron dense deposits within the basal lamina of the renal glomeruli are observed frequently. 4. Amount of the microfilaments in the mesangial cells and the mesangial matrices are increasing during aging. 5. The thicknesses of the basal laminae of the Bowman's capsule are increasing during aging. 6. After twenty four months, the proximal tubular cell-like parietal cells with well developed microvilli are observed frequently. From the above results, it was suggested that the renal glomerulus matures structurally in five weeks, and the function of the glomerulus is suppressed after eighteen months.
Garlic occupies a special position among the many foods of vegetable origin because it is the sole food for Koreans during the their lives. And vitamin A has been ingested by forms of food or additives. Cadmium has been described as one of the most dangerous trace elements in the food and environment of man and livestocks. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of cadmium-induced changes in gene expression , ie. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and damage for food hygiene. He acute and chronic combine effects of cadmium (Cd, CdCl2 20mg/kg), garlic oil(Dds: diallyl disulfide 50mg/kg, 3 times a week) and vitamin A(Ra: retinol acetate 50,000 IU/kg, 3 times a week) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression histopathological and electron microscopical examinations. The results of the study are as follows ; 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in tissue were not changed by the simultaneous adminstration of diallyl disufide or retinol acetate. 2. ALT(alanine aminotransferase) , AST(aspartate aminotransferase), glucose, BUN (blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly hanged by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Histopathological changes in cadmium treated rats were appeared at 8 weeks age treatment in kidneys. Homogenous eosinophilic material was accumulated in cortical and collecting tubular lumens at 16 weeks. Degenerated or necrotized tubular cells were observed in cortex and medulla. Degenerated seminiferous tubules and homogeneous eosinophilic material was seen in interstitial tissue of rat treated with cadmium for 16 weeks. Calcium deposits were seen in degenerated seminiferous tubules and the tubules showed severe calcification of rat treated with cadmium for 16 weeks. Electron microscope changes in kidney were observed in rats treated with CdCl2 20 mg/kg. Proximal convoluted tubule cells showed selling of cytoplasm and narrow lumen. Capillary endothelial cells showed cytoplasmic vacuoles and swelling. Degenerated epithelial cells were accumulated in tubular lumen of kidney. 4. Enhanced synthesis of 70 KDa relateve molecular mass proteins were detected in 2 hours after cadmium, exposure, with maximum activity occurring at 8~48 hours. Induction of HSP 70 was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that HSP70 induction by the cadmium treatment was a rapid reaction to indicated the exposure of xenobiotics, and retinol acetate reduced the cadmium induced nephrotoxicity.
Renal failure syndrome in wild mammals is infrequently reported. Muskrat (Ondatra zibethicus) is a medium-sized rodent known to carry many diseases but rarely exhibiting renal failure. A six-month old female muskrat was submitted to our laboratory for pathological diagnosis, and necropsy revealed severe renal damage with sand-like lithiasis in the ureter, renal calculi, and hydronephrosis. All major organs, including the cerebrum, also showed systemic hemorrhage and calcification which may have been due to uremia induced by renal failure. Histopathologically, necrosis and microcalcification were detected in the renal cortex and the medulla, especially in the proximal convoluted tubules and collecting ducts of the kidney. Significant hyalinization of the glomeruli was also observed, and this suggested chronic nephritis. These findings would support mycotoxic effects, particularly on the kidney. Moreover, infiltration of neutrophils and mononuclear cells was observed in the lung and of plasma cells in the spleen. The definitive cause of the toxic effects in this case of muskrat renal failure could be attributed to contaminated food.
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