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  • 1
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of triiodo-L-thyronine (T3) treatment (15 μg/100 g body weight daily for 10 days) on the activity of mitochondrial α-glycerophosphate dehydrogenase (GPOX) in different nephron segments of the male rat was investigated by a histochemical staining method. The study showed marked segmental differences regarding the response to T3-treatment: 1. The first two proximal segments were unstained in the control rats andintensely stained following treatment. 2. The third proximal segments, the thick ascending limbs of Henle's loop and the distal convoluted tubules showed a strong or moderate reaction in controls and amoderate increase after T3-treatment. 3. The high activity of collecting ducts in the cordex and outer zone of the medulla in controls wasslightly increased by treatment. 4. Faintly reacting glomeruli and negative thin limbs of Henle's loop and collecting ducts in the inner medulla (papilla) wereunaffected by T3-treatment. The results are discussed in relation to biochemical and physiological data.
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  • 2
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The segmentation of the proximal tubules of the male rat kidney was studied by means of enzyme histochemical reactions. Soluble oxidoreductases (glucose 6-phosphate dehydrogenase, α-glycerophosphate dehydrogenase, 3α-hydroxysteroid dehydrogenase, NAD- and NADP-dependent isocitrate dehydrogenases, NAD-dependent malate dehydrogenase, NADP-dependent, decarboxylating malate dehydrogenase, uridine diphosphate glucose dehydrogenase) were demonstrated using methods which reduce enzyme diffusion (incubating in presence of polyvinyl alcohol) and eliminate interference from tissue tetrazolium reductases. Less soluble or insoluble enzymes (glucose 6-phosphatase, β-hydroxybutyrate dehydrogenase, succinate dehydrogenase and tetrazolium reductases) were demonstrated by incubation in conventional watery media. Segmental differences were observed in respect to all enzymes studied, and most reactions clearly visualized the three segments known to exist from ultrastructural as well as previous histochemical studies: The pars convoluta includes the first (P1) and most of the second (P2) segment. The transition to the third segment (P3) is in the beginning of the pars recta. Also these reactions revealed a difference between the first part of the P3, which runs through the cortex in the medullary rays, and the terminal part transversing the outer stripe of the medulla. In most instances intensity of reaction decreased in the last portion of the P3. A number of the enzymes studied were mainly or solely localized to the P3 (glucose 6-phosphate dehydrogenase, α-glycerophosphate dehydrogenases, β-hydroxybutyrate dehydrogenase, 3α-hydroxysteroid dehydrogenase, decarboxylating malate dehydrogenase and uridine diphosphate glucose dehydrogenase). Some possible functional implications of the findings are discussed.
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  • 3
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The histochemical localization of lactic dehydrogenase (LDH) activity and the prevailing type of isoenzyme in different segments of the nephron in male and female rats are described. Polyvinyl alcohol was added to the incubation medium in order to reduce enzyme diffusion. Localization of the reaction product was further improved by the use of a high concentration of Nitro BT (and of PMS and NAD). The three segments of the proximal tubules exhibited clearly different staining patterns. In glomeruli, terminal parts of the third proximal segments, thin limbs of Henle, and collecting ducts M subunits of LDH predominated, whereas in the remaining tubular segments H subunits prevailed. Sex differences were observed in respect to the LDH reaction in the proximal tubules, but not in the rest of the nephron. The localization of α-hydroxy acid oxidase was also investigated, as this enzyme oxidizes lactate and therefore contributes to the reaction. Under certain conditions, i.e. high substrate concentration, the activity of α-hydroxy acid oxidase was negligible in comparison with that of LDH.
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  • 4
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The segmentation of the proximal tubules in the kidney of the female rat was studied by means of enzyme histochemical reactions and the results compared with those observed in the male and recently described by Jacobsen and Jørgensen (1973a). Reactions were performed for the following soluble, coenzyme-dependent oxido-reductases: glucose 6-phosphate dehydrogenase, α-glycerophosphate dehydrogenase, 3 α-hydroxysteroid dehydrogenase, NAD-as well as NADP-dependent isocitrate dehydrogenases, NAD-dependent malate dehydrogenase, NADP-dependent, decarboxylating malate dehydrogenase, uridine diphosphate glucose dehydrogenase. Measures were taken to reduce enzyme diffusion and eliminate interference from tissue tetrazolium reductases. Furthermore, reactions were performed for a number of less soluble or insoluble enzymes: glucose 6-phosphatase, mitochondrial α-glycerophosphate dehydrogenase, β-hydroxybutyrate dehydrogenase, succinate dehydrogenase and tetrazolium reductases. In the proximal tubules of the female rat all enzymes studied—except β-hydroxybutyrate dehydrogenase—showed segmental differences, most of them clearly revealing three segments. Sex differences were found concerning all enzymes except uridine diphosphate glucose dehydrogenase and NADP-dependent isocitrate dehydrogenase. The most pronounced sex-related differences were seen in the third segment in which part the male rat showed highest activity in respect to tetrazolium reductases, NAD-dependent isocitrate dehydrogenase, succinate dehydrogenase, β-hydroxybutyrate dehydrogenase, 3 α-hydroxysteroid dehydrogenase and glucose 6-phosphate dehydrogenase and the female in respect to glucose 6-phosphatase, α-glycerophosphate dehydrogenases, and NADP-dependent, decarboxylating malate dehydrogenase. A few of the enzymes exhibited minor sex differences in the first two segments.
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  • 5
    ISSN: 1432-0428
    Keywords: Key words Diabetes mellitus ; kidneys ; thick ascending limb of the loop of Henle ; Tamm ; Horsfall glycoprotein.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Tamm–Horsfall glycoprotein, present only in the kidney thick ascending limb of Henle's loop, was studied here in streptozotocin diabetic rats. Tamm–Horsfall glycoprotein mRNA in situ hybridization was performed on snap-frozen left kidneys; the right kidneys were perfusion-fixed with 4 % paraformaldehyde and embedded either in paraffin, for Tamm–Horsfall glycoprotein immunohistochemistry, or in Epon for stereologic measurements. The length of the thick ascending limb of Henle's loop and the amount of glycogen were measured and the ultrastructure of the cells was evaluated. Urinary excretion of Tamm–Horsfall glycoprotein, calcium, magnesium and albumin was measured. After 10 and 50 days' duration of diabetes, kidney weight increased 20 and 41 %, respectively and the length of the thick ascending limb of Henle's loop increased 28 and 56 %, respectively, compared with controls. Substantial glycogen accumulations were present in the thick ascending limb of Henle's loop, and electron microscopy revealed a significant decrease in organelles and basolateral membranes. After 10 and 50 days' duration of diabetes, in situ hybridization of Tamm–Horsfall glycoprotein mRNA revealed a fourfold decrease, and the immunostaining for Tamm–Horsfall glycoprotein showed a threefold decrease as measured by densitometry. However, urinary Tamm–Horsfall glycoprotein excretion rate was increased fivefold and urinary concentration about twofold. Urinary calcium excretion increased threefold and magnesium twofold, but urinary albumin excretion was not significantly increased. The increased amount of Tamm–Horsfall glycoprotein, calcium and magnesium in the urine in diabetes occurs here concomitant with severe cellular damage in the thick ascending limb of Henle's loop. [Diabetologia (1995) 38: 525–535]
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  • 6
    ISSN: 1432-0428
    Keywords: Diabetes mellitus ; kidneys ; thick ascending limb of the loop of Henle ; Tamm-Horsfall glycoprotein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Tamm-Horsfall glycoprotein, present only in the kidney thick ascending limb of Henle's loop, was studied here in streptozotocin diabetic rats. Tamm-Horsfall glycoprotein mRNA in situ hybridization was performed on snap-frozen left kidneys; the right kidneys were perfusion-fixed with 4% paraformaldehyde and embedded either in paraffin, for Tamm-Horsfall glycoprotein immunohistochemistry, or in Epon for stereologic measurements. The length of the thick ascending limb of Henle's loop and the amount of glycogen were measured and the ultrastructure of the cells was evaluated. Urinary excretion of Tamm-Horsfall glycoprotein, calcium, magnesium and albumin was measured. After 10 and 50 days' duration of diabetes, kidney weight increased 20 and 41%, respectively and the length of the thick ascending limb of Henle's loop increased 28 and 56%, respectively, compared with controls. Substantial glycogen accumulations were present in the thick ascending limb of Henle's loop, and electron microscopy revealed a significant decrease in organelles and basolateral membranes. After 10 and 50 days' duration of diabetes, in situ hybridization of Tamm-Horsfall glycoprotein mRNA revealed a fourfold decrease, and the immunostaining for Tamm-Horsfall glycoprotein showed a threefold decrease as measured by densitometry. However, urinary Tamm-Horsfall glycoprotein excretion rate was increased fivefold and urinary concentration about twofold. Urinary calcium excretion increased threefold and magnesium twofold, but urinary albumin excretion was not significantly increased. The increased amount of Tamm-Horsfall glycoprotein, calcium and magnesium in the urine in diabetes occurs here concomitant with severe cellular damage in the thick ascending limb of Henle's loop.
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  • 7
    ISSN: 1432-069X
    Keywords: D-Amino acid oxidase ; Epidermal cells in culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The occurrence of the enzyme D-amino acid oxidase in human epidermis and cultured epidermal cells was investigated. When explant cultures of human epidermis were cultured in a medium containing D-valine instead of L-valine and supplemented with undialyzed serum, good growth of both epithelial cells and fibroblasts was observed. However, when the serum was dialyzed neither cell type could be cultured in D-valine medium indicating the absence of D-amino acid oxidase in both cell types. When epithelial cultures initiated in L-valine medium were changed to D-valine medium after 1–2 weeks, growth stopped immediately, and the epithelial cells showed signs of extensive degeneration, indicating that skin epithelial cells have a very low endogenous pool of L-valine. When these cultures were re-fed L-valine medium after 2 weeks in D-valine medium, this resulted within a few days in the reappearance of epithelial outgrowth. The activity of D-amino acid oxidase in human epidermal cells was further studied by histochemistry and in homogenates of epidermis and cultured epidermal cells. Whereas high activity of histidase was observed in the epidermal cells, no activity of D-amino acid oxidase could be detected. This report shows that D-amino acid oxidase does not serve as a marker enzyme for epithelial cells in general as previous tissue culture studies might indicate. Human skin epithelial cells do not contain detectable amounts of D-amino acid oxidase activity, and therefore, D-valine medium is not suitable for selection of growth of skin epithelial cells in culture.
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  • 8
    ISSN: 1432-2218
    Keywords: Key words: Colon — Laparoscopic colectomy — Surgical technique — Aseptic colon resection — Animal study
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: We evaluated a new aseptic method for laparoscopic left colon resection in terms of technical feasibility and outcome. Methods: Ten pigs were operated on under general anesthesia. Pre- and postoperative body weight, stools, behavior, and need for analgesics were recorded. Fourteen days later, the animals were killed. At autopsy, the degree of intraabdominal adhesions was noted. The anastomoses were sent for histological examination. The entire procedure was performed intracorporeally, and no antibiotics were given. After division of the mesocolon, the segment to be resected was invaginated down through the colon. This was facilitated by a custom-made instrument that was introduced into the bowel via the anus; it consisted of a pull-out device and a modified diathermy wire. The anastomosis was completed at the invagination fold by a row of hernia staples that were covered by an interrupted suture. Then the invaginated bowel was transected by the diathermy wire and delivered through the anus. Results: One animal was killed before completion of the operation because of a colonic perforation. The remaining nine animals had an uneventful and rapid recovery. They ate from the 1st postoperative day and gained weight rapidly. Stools were normal after 2 days (median), and normal behaviour was noted in all animals from the 1st postoperative day. At the postmortem examination, intraabdominal adhesions were observed in two animals. In one case, the adhesions extended from a hematoma in the mesentery to the abdominal wall. There were no adhesions to the anastomosis or the colon. In the other case, the anastomosis adhered to the right uterine tube and a loop of small intestines. Conclusions: The method is technically feasible, but a modification is suggested for cases where the invagination is impossible. Recovery after the operation is rapid.
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  • 9
    ISSN: 1573-2568
    Keywords: achalasia ; eosinophils ; cytotoxicity ; VIP ; substance P ; acetylcholinesterase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We present a case of secondary achalasia due to an adenocarcinoma of the stomach with no tumor infiltration of the esophagus. Immunohistochemical staining revealed a massive infiltration of activated eosinophils in the muscularis of the esophagus with secretion of the highly cytotoxic and neurotoxic eosinophil cationic protein (ECP). Immunohistochemical staining for the neuropeptides VIP and substance P, as well as the histochemical demonstration of AChE, revealed a nearly total absence of all three neurotransmitters/modulators compared to control. The hypothesis is advanced that eosinophil neurotoxicity is the cause of secondary achalasia.
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  • 10
    ISSN: 1432-0878
    Keywords: Kidney ; Lithium ; Collecting ducts ; Intercalated cells ; Vasopressin ; Diabetes insipidus ; Wistar rats ; Long Evans rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rats were given a lithium-containing diet (40 mmol/kg) to Study the effect of lithium on the structure of collecting ducts from the inner stripe of the outer medulla. The results show that there is a significant increase in the volume density of collecting ducts already after one week on this diet. The volume density of both intercalated and principal cells increases, whereas the volume density of mitochondria in the cytoplasm increases in the intercalated cells only. The increased volume of both principal and intercalated cells seems to be part of a general hyperplasia and hyperactivity of the collecting duct, which may in some way be related to the effects of lithium on vasopressinmediated water transport. The specific changes in the intercalated cells may be a consequence of the effects of lithium on distal nephron potassium and hydrogen ion transport in the distal nephron.
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