Morphological,histochemical, and biochemical studies on the gut ofHaemonchus contortus Rud., 1803

Summary A study has been made on the structure and chemical composition of the gut ofHaemonchus contortus (Rud., 1803). The oesophagus has typically a triradiate, cuticle-lined lumen. The intestinal epithelium is provided with a well-developed brush border which contains periodic acid-Schiffpositive mucoproteins. The intestinal epithelium stores glycogen and lipids. It stains diffusely for phospholipids and general proteins and also for terminal-NH₂ group. The presence of Fe²⁺ and Fe³⁺ containing pigments and activities of acid and alkaline phosphatases, glucose-6-phosphatase, and 5-nucleotidase have been observed in the intestinal epithelium. Biochemically pH optimum for intestinal acid phosphatase has been found to be 4.8. The brush border shows positive reactions for acid phosphatase and glucose-6-phosphatase, and negative reactions for alkaline phosphatase and 5-nucleotidase. The presence of enzymes in the brush border is related to extracellular digestion and absorption of nutrients.     

Effect of thyroid hormone on blood and liver glucose-6-phosphatase activity

Summary Experimental thyrotoxicosis of mice and rats causes a significant (P<0.01 and P<0.001) rise in hepatic glucose-6-phosphatase activity, but did not affect that of blood plasma. Hepatic alkaline phosphatase activity was raised in thyrotoxic rats, while plasma alkaline phosphatase activity was lowered.(Presented by Member of the Academy of Medical Sciences USSR S. E. Severin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 55, No. 6, pp. 59–61, June, 1963     

The localization of the Na+−K+-ATPase in the cells of rat kidney cortex

Summary Plasma membrane fractions of rat kidney cortex were subdivided by centrifugation on a continuous and a discontinuous sucrose gradient and by carrier free continuous electrophoresis. In the different fractions the activity of alkaline phosphatase and aminopeptidase, enzymes which are present in the brushborder membrane, as well as Mg⁺⁺-ATPase, Na⁺–K⁺-ATPase, 5-nucleotidase, acid phosphatase and glucose-6-phosphatase were determined.The distribution of alkaline phosphatase, aminopeptidase and 5-nucleotidase is identical, indicating the localization of these enzymes in the brushborder membrane. Na⁺–K⁺-ATPase does not show an identical distribution with any of the enzymes studied.Using carrier free continuous electrophoresis fractions can be obtained which are enriched in alkaline phosphatase by a factor of 15 when compared to the cortex homogenate, whereas the specific activity of Na⁺–K⁺-ATPase is reduced to one third of the starting material. On the other hand fractions can be isolated in which the specific activity of Na⁺–K⁺-ATPase is 16 times higher than in the homogenate. No enrichment of alkaline phosphatase occurs in these fractions.It is therefore concluded that the Na⁺–K⁺-ATPase is not present in the brushborder membrane nor in the lysosomes or endoplasmatic reticulum. The most probable localization of the Na⁺–K⁺-ATPase are the basal infoldings of the plasma membranes of the cells.A preliminary report has been published by Kinneet al. [28, 29].Major part of this work was done by J. E. Schmitz for his degree of M. D.     

Protective effect of cysteine and methylprednisolone in lead acetate-endotoxin induced shock

To delineate possible mechanisms of lead acetate enhancement to endotoxin shock, liver parenchymal, and Kupffer cell function were evaluated in lead acetate-treated rats receiving low doses of S. enteritidis endotoxin. Additionally, the influence of methylprednisolone and cysteine, agents which have been demonstrated to prevent lead-induced endotoxin lethality was ascertained. Rats receiving lead acetate demonstrated profound hyperreactivity to endotoxin as denoted by increased mortality, impairment of bromsulphalein (BSP) removal, hypoglycemia, and increased blood lactate and pyruvate. Plasma activity of glutamic pyruvic transaminase (GPT), glutamic oxalacetic transaminase (GOT), and alkaline phosphatase was strikingly elevated. Serum activity of beta-glucuronidase and plasma acid phosphatase was also elevated. Methylprednisolone or cysteine treated rats were essentially refractory to the lethal effects of lead acetate and endotoxin. The enhanced survival of lead-endotoxin treated rats following administration of methylprednisolone or cysteine was associated with reduced impairment in BSP removal, a normal blood glucose, and significant reductions in blood lactate and pyruvate. Plasma activity of GPT, GOT, and alkaline phosphatase was also reduced. Slight reductions were observed in plasma activity of acid phosphatase although serum beta-glucuronidase activity was not significantly altered. Phagocytic activity, as measured by hepatic uptake of gelatinized RE test lipid emulsion, indicated a hypofunctional state of Kupffer cells in lead acetate-endotoxin treated rats. Since neither methylprednisolone, nor cysteine treatment modified the hypophagocytic state, Kupffer cell phagocytic dysfunction does not appear to play a major role in enhanced endotoxin susceptibility of lead acetate treated rats. The maintenance of parenchymal cell function in methylprednisolone or cysteine treated rats injected with lead and endotoxin supports the hypothesis of a contributory role for hepatic parenchymal cells in the pathophysiology of lead-endotoxin interaction.     

Histochemische Untersuchungen ischÄmisch bedingter Nebennierenrinden-Nekrosen bei Ratten

Zusammenfassung Die GefÄ\e der linken Nebenniere von Ratten wurden 60, 90 oder 120 min lang abgeklemmt und das Organ 1, 2, 4, 6, 12 und 24 Std sowie 4, 7, 10, 14, 21 und 28 Tage nach Absetzen der Unterbindung histochemisch untersucht. 2 Std nach Beendigung der Abklemmung ist im Bereich der dabei auftretenden Infarkte eine betrÄchtliche Abnahme der histochemisch nachweisbaren basophilen Substanz des Cytoplasmas und ein Nachlassen der ⁵-3-Hydroxysteroid-Dehydrogenase sowie der alkalischen Phosphatase-Reaktion der Capillaren feststellbar, nach 4 Std eine AktivitÄtsverminderung — und nach 12 Std völliger Ausfall — der übrigen untersuchten Fermente (Succindehydrogenase, saure Phosphatase, nichtspezifische Esterase). Das histochemische Bild des überlebenden Parenchyms unterscheidet sich nicht von dem der Kontrollen. Die von den überlebenden Parenchyminseln der Zona glomerulosa und der tieferen Zonen ausgehenden Regenerate zeigen die gleiche FermentaktivitÄt wie die Zellen der intakten Nebenniere.

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Histochemical studies of adrenal infarcts in rats produced by temporary ischemia

Summary The vessels of the left adrenal of rats were clamped for 60, 90, and 120 minutes. The histochemical alterations were studied and compared with the histological changes 1, 2, 4, 6, 12, and 24 hours and 4, 7, 10, 14, 21 and 28 days after the clamps were released. Two hours after the temporary ischemia in the region of the infarcts considerable diminution of the basophilic substance of the cytoplasm and decrease in the activity of the ⁵-3- hydroxysteroid dehydrogenase could be detected, and the alkaline phosphatase of the capillaries decreased as well. Within four hours thereafter a fall in the activity (and after 12 hours complete disappearance) of the other enzymes examined (succinic dehydrogenase, acid phosphatase, non-specific esterase) could be observed. The histochemical picture of the surviving parenchyma could not be distinguished from that of the controls. The activity of the enzymes studied in the newly formed cells of the regenerating parts was similar to that of the cells of the intact adrenal cortex.

     

Serum biochemical assessment of hepatic and renal functions of rats during oral exposure to glyphosate with zinc

A subchronic toxicity study was carried out to assess hepatic and renal functions of rats during oral exposure to glyphosate with zinc for the period of 8 weeks. Forty-eight Wistar rats used for the study were randomized into six groups of eight Wistar rats each, and each group had equal number of male and female Wistar rats. The Wistar rats administered with distilled water at 2 ml/kg body weight served as the control group (DW); others were administered with zinc at 50 mg/kg body weight (Z) group, glyphosate at 375 mg/kg body weight (G) group, a combination of zinc and glyphosate at 50 and 375 mg/kg body weight, respectively (Z?+?G), group, glyphosate at 14.4 mg/kg body weight (GC) group, and a combination of zinc and glyphosate at 50 and 14.4 mg/kg body weight, respectively (Z?+?GC), group. At the end of the study, blood samples were collected from each rats; from which, sera samples were obtained and assayed for total protein, albumin, alanine and aspartate aminotransferases, alkaline phosphatase, Na⁺, K⁺, Cl^?, \( \mathrm{HCO}_3^{-} \) , Ca²⁺, \( \mathrm{PO}_4^{-} \) , urea and creatinine using autoanalyzer, and globulin was calculated. The albumin concentration was significantly high (p?<?0.05) in GC group compared to DW group, and this change was ameliorated following supplementation with zinc. The total protein and globulin concentrations did not differ significantly between the groups (p?>?0.05), and the relative changes were ameliorated by supplementation with zinc. The alkaline phosphatase activity was relatively low in GC group; however, supplementation with zinc in Z?+?GC group made it to be significantly high (p?<?0.05) compared to GC group. The alanine and aspartate aminotransferases in G and GC groups were relatively high compared to DW group, which were ameliorated by supplementation with zinc. The relatively low Ca²⁺ concentration in G and GC groups compared to DW were ameliorated in Z?+?G group, and it was significantly high in Z?+?GC group at p?<?0.01 compared to DW, p?<?0.001 compared to G and GC groups and p?<?0.05 compared to Z?+?G group. There were only slight changes in the electrolytes concentrations (Na⁺, K⁺, Cl^?, \( \mathrm{HCO}_3^{-} \) and \( \mathrm{PO}_4^{-} \) ), which were differentially ameliorated by zinc supplementation. The reasons for the various changes recorded were discussed. It was concluded that subchronic oral exposure to glyphosate caused both hepatic and renal functions toxicity in rats, which were ameliorated by zinc supplementation.     

Reactivity of capillaries and pyramidal neurons in rat cortex under conditions of acutely reduced circulation

A study of capillaries and pyramidal neurons of the cortex in white rats subjected to ligation of the common carotid artery reveals that acute reduction of the circulation is characterized by a decreased activity of alkaline phosphatase and Na, K-ATPase as well as a reduced number of capillaries marked with these enzymes, and a lower activity of butyrylcholine esterase. A decreased activity of cytochrome oxidase in neurons suggests the existence of intracellular compensatory-adaptive mechanisms. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 5, pp. 558–560, May, 1994 Presented by O. S. Adrianov, Member of the Russian Academy of Medical Sciences     

Visualization of capillaries in skeletal muscle by the ATPase reaction

Summary A simple and reliable technique for the visualization of capillaries in skeletal muscle of dogs, guinea pigs and rats is described. 10–20 μm frozen sections were incubated in a medium containing Ca²⁺ and ATP following acid preincubation (pH 3.8–4.2). The capillaries stained in black and were readily seen surrounding the muscle fibers. Serial sections were also treated for alkaline phosphatase. Values for capillary density using both methods were not different. Supported by USPHS NIH Grant HL 12679-HL 18145 Supported by a fellowship from the Rockefeller Fndn     

Calcitonin treatment of immobilization osteoporosis in rats

We studied changes in bone mass induced by immobilization and the ability of salmon calcitonin to inhibit immobilization osteoporosis in rat. The bone mass of the immobilized hind leg of rat was compared with the contralateral non-treated leg. Neurectomy and cast immobilization reduced the bone mineral mass to an equal extent. However, the dose-response of calcitonin was different with these immobilization techniques. Calcitonin 15 IU kg^-1 administered once daily reduced bone ash weight difference significantly after 2 weeks' neurectomy (P < 0.001). This had no significant effect on the bone loss induced by cast immobilization, but the dose had to be delivered as two injections given every 12 h. Two weeks' immobilization decreased the incorporation of ⁴⁵Ca into bones. Calcitonin could not prevent this. However, calcitonin tended to inhibit the overall incorporation of 45Ca into bones in immobilized rats but yet had no effect on ⁴⁵Ca incorporation in non-immobilized rats. Immobilization decreased serum alkaline phosphatase activity in cast-immobilized animals. Neurectomy did not change serum alkaline phosphatase activity from a sham operation level. The tartrate-resistant acid phosphatase to total acid phosphatase ratio in the serum increased significantly in neurectomized rats and in cast-immobilized calcitonin-treated rats.     

The effect of long-term administration of α1-blocker prazosin on capillary density in cardiac and skeletal muscle

The effect of prazosin on heart and muscle blood flow and capillary density was studied in rats. In acute experiments, ₁-blocker prazosin almost trebled blood flow in fast skeletal muscles [tibialis anterior (TA) and extensor digitorum longus (EDL)], but did not affect coronary flow when infused i.v. at a dose of 0.5 g · ml^–1 · min^–1 for 30 min. Prazosin in an equivalent dose was then given orally over a period of 5 weeks to investigate its effect on capillarisation in heart and skeletal muscle. Capillary density (CD, capillaries · mm^–2), estimated in frozen sections stained for alkaline phosphatase, was similar in the hearts of prazosin-treated and control rats. Capillary/fibre ratio in skeletal muscles increased from 1.52±0.019 in control EDL to 1.69±0.01 (P<0.001) and from 1.56±0.04 in control TA to 2.16±0.04 (P<0.001). In TA, the increase was greater than in EDL both in the glycolytic periphery (from 1.30±0.13 to 1.75±0.11, P<0.025) and the oxidative core of the muscle (from 1.837±0.14 to 2.51±0.12, P<0.005). Unilateral crush of the lateral peroneal nerve and subsequent reinnervation over the next 7 weeks resulted in redistribution of fibre types from a typical mosaic pattern into groups composed of fibres of similar oxidative capacity. Capillary density as well as capillary/fibre ratio in purely glycolytic areas was lower when compared to supply of glycolytic fibres in normal muscles. Oral administration of prazosin over the whole period of reinnervation not only maintained the original level of capillarity associated with fast glycolytic fibres in control muscles, but considerably increased it. Thus long-term prazosin administration not only causes an increase in capillary supply in normal muscles but also prevents loss of capillaries during reinnervation. The fact that it only increases capillarisation in tissues where it increases flow further supports the hypothesis that capillary growth can be initiated by mechanical factors connected with high blood flow.     

Chosphatases of the human placenta

Summary Acid and alkaline phosphatase activity has been examined in chorionic and placental tissues of 123 women in the 4–5th to the 40th week of pregnancy, by the method of Gomori. Both enzymes could be detected over the whole duration of pregnancy. Most of the villi of preparations from early pregnancy showed trace or low acid phosphatase activity, and the proportion of such villi increased as pregnancy progressed. At the same time, there was a gradual rise in the acid and alkaline phosphatase activity of the structures, to a maximum at term. Alkaline phosphatase was present in the nuclei, but more abundantly in the cytoplasm of the syncytium; it was absent from the Langhans cells. Acid phosphatase was concentrated mainly in the nuclei, and was more widely distributed among the placental tissues than was alkaline phosphatase. The trophoblast showed the highest acid and alkaline phosphatase activities. The results reported are not in agreement with those of Thomsen.(Presented by Member AMN SSSR A. V. Lebedinskii) Translated from Byulleten Éksperimental'noi Biologii i Meditsiny, Vol. 56, No. 8, pp. 64–68, August, 1963     

Response of lung enzyme activities in rabbits following short-term exposure ton-hexane: Correlation between morphological and biochemical changes

The activity of lactatedehydrogenase, -glucuronidase, glucose-6-phosphate dehydrogenase, acid and alkaline phosphatase was studied in lung homogenate from New Zealand rabbits exposed to 3000 p.p.m. ofn-hexane 8 h per day for 8 days or filtered air.In hydrocarbon-treated animals all enzymes examined, except alkaline phosphatase, were markedly increased.The biochemical changes correlated well with the morphological changes and the results of cytological evaluation of bronchopulmonary lavage. It is suggested that high values in lung lysosomal enzymes from treated rabbits reflect the acute inflammation whilst the increase in lung glueose-6-phosphate dehydrogenase may depend upon reparative process subsequent ton-hexane-induced lung damage.     

Whole body59Fe-elimination rates and corresponding blood losses in patients with factitious anemia induced by self-blood letting

Summary Factitious anemia induced by self-bloodletting is a rare cause of chronic haemorrhagic iron deficiency anemia. Within a year two cases from Southern Germany were identified and quantitated for blood losses by whole body counting of retained iron. Whereas the 2 patients did not lose much blood (6–10 ml blood/day equivalent to 0.12–0.34%⁵⁹Fe-whole body elimination/day) during their observation within the hospital their blood losses increased to 41–187 ml/day (=whole body⁵⁹Fe-elimination rates of 0.8 to 6.1%/day) upon release from the hospital. Although they lost about 5 liter blood in 50 days faecal occult blood tests were always negative and the daily faecal and urinary⁵⁹Fe-excretion was with 0.018–0.031%/day within the normal range. Haemoglobin and erythrocyte levels were stable during the periods of clinical observation when the whole body⁵⁹Fe-elimination rates were only just above the normal range (0.12–0.21%/d). These values dropped however to very low values during or following periods of extremely elevated whole body⁵⁹Fe-elimination rates (1–6%/d=46–187 ml blood loss/d) immediately after release of the patients from the hospital. One patient received more than 400 blood and erythrocyte infusions (84 g iron) and absorbed about 8 g food iron within 10 years. Nevertheless she was always severely anemic (5 g% haemoglobin) and had no storage iron in the tissues. She must have lost about 92 g iron in 10 years (= 529 liter blood) or 25 mg Fe/d (= 145 ml blood/d) by self bloodletting avoiding the otherwise extreme generalized transfusion haemosiderosis. Blood losses of up to 145 ml/d (= 50 mg Fe/d) can be compensated by oral ferrous iron therapy with 4×50=200 mg Fe²⁺/d. The patients did however refuse to take oral or parenteral iron and insisted in one case on regular blood transfusions twice per week.Two more probable cases of factitious anemia from Northern Germany were Subsequently observed and partially investigated (cases 3 and 4).With the technical assistance of Fatima Iagi     

Effect of reduced red cell “deformability” on flow velocity in capillaries of rat mesentery

A graded reduction of deformability of red blood cells (RBC's) of rats was obtained by treatment with the SH-oxidizing agent, diamide. Rigidified RBC's were injected into rats by isovolemic exchange against 60% of the native RBC's and RBC flow velocities in capillaries of rat mesentery measured.At normal mean arterial pressure RBC flow velocity decreases by 29% in rats receiving cells rigidified with 0.5 mmol·l^–1 diamide. Surprisingly a further rigidification of erythrocytes by 1.5 mmol·l^–1 diamide results in a decrease of flow by only 15%. During hypotension RBC flow velocities dropped precipitously to 8±15% for the 0.5 nmol·l^–1 and to 2±6% for the 1.5 mmol·l^–1 diamide group compared to velocities during normotension. By microscopy we observed a stop of flow in many vessels.This result outlines the importance of a normal red cell deformability for the maintenance of sufficient perfusion of the microcirculation, in particular at low blood pressure gradients.Supported by Deutsche Forschungsgemeinschaft (SFB 109 C5)Presented in part at the 50th meeting of the Deutsche Physiologische Gesellschaft, October 3–6, 1978, Göttingen, and at the 2nd World Congress of Microcirculation, July 1978     

S35-methionine uptake by proteins of the structural elements of tissue cells in irradiated rats

Summary The rapidity of inclusion of S³⁵-methionine into the cells of various tissues was studied in rats in acute radiation sickness. Inclusion of S³⁵-methionine was studied in the proteins of cellular nuclei (desoxynucleoprotein, acid and residual), mitachondria and microsomes (water-soluble ribonucleoproteins, lipoproteins) and hyaloplasm of the liver and of the mucous membrane of the small intestine.It was established that x-ray irradiation causes increased inclusion of radioactive sulfur methionine into all proteins of the cellular nuclei of the liver, as well as in the acid protein of the mucous membrane of the small intestine. Inclusion of S³⁵-methionine into the desoxyribonucleoprotein and residual protein of the nuclei in the mucous membrane of the small intestine does not change in comparison with the same in normal conditions.Decreased inclusion of radioactive sulfur methionine is noted in cytoplasmic proteins of the liver and in the mucosa of the small intestine. This decrease is especially pronounced in ribonucleoproteins of mitochondria and microsomes.Submitted by Active Member of AMN SSSR V. N. Otekhovich     

Moderate exercise training promotes adaptations in coronary blood flow and adenosine production in normotensive rats

OBJECTIVES:

Aerobic exercise training prevents cardiovascular risks. Regular exercise promotes functional and structural adaptations that are associated with several cardiovascular benefits. The aim of this study is to investigate the effects of swimming training on coronary blood flow, adenosine production and cardiac capillaries in normotensive rats.

METHODS:

Wistar rats were randomly divided into two groups: control (C) and trained (T). An exercise protocol was performed for 10 weeks and 60 min/day with a tail overload of 5% bodyweight. Coronary blood flow was quantified with a color microsphere technique, and cardiac capillaries were quantified using light microscopy. Adenine nucleotide hydrolysis was evaluated by enzymatic activity, and protein expression was evaluated by western blot. The results are presented as the means ± SEMs (p<0.05).

RESULTS:

Exercise training increased the coronary blood flow and the myocardial capillary-to-fiber ratio. Moreover, the circulating and cardiac extracellular adenine nucleotide hydrolysis was higher in the trained rats than in the sedentary rats due to the increased activity and protein expression of enzymes, such as E-NTPDase and 5′-nucleotidase.

CONCLUSIONS:

Swimming training increases coronary blood flow, number of cardiac capillaries, and adenine nucleotide hydrolysis. Increased adenosine production may be an important contributor to the enhanced coronary blood flow and angiogenesis that were observed in the exercise-trained rats; collectively, these results suggest improved myocardial perfusion.     

Modulation of colon-Derived Experimental Hepatic Metastasis by murine Nonfarenchymal liver Cells

Although numerous animal tumor models have been used to study colon carcinoma, few display hepatic metastasis. C57B1/6Ros mice inoculated with liver-derived murine colon adenocarcinoma MCA-38 in the ileocolic vein develop distinct hepatic foci within 21 days and survive an average of 35 days. Furthermore, ¹¹¹In-labeled LD-MCA-38 tumor cells were rapidly taken up by the liver within 60 min and 73% of the label remained in the liver after 24 h. Isolated nonparenchymal liver cells from untreated mice displayed little cytotoxicity against freshly excised ⁵¹Cr-labeled MCA-38 cells but did inhibit tumor growth in vitro as measured by inhibition of ³H-thymidine incorporation. Treatment with anti-asialo-G_M1 decreased the lifespan of MCA-38 tumor bearing mice suggesting that asialo-G_M1 positive cells in the liver may inhibit tumor growth in vivo. Nonparenchymal liver cells from mice treated with polyinosinic-polycytidylic acid showed augmented cytotoxic and cytostatic activity against LD-MCA-38 tumor cells in vitro. Polyinosinic-polycytidylic acid treatment also significantly increased the lifespan of MCA-38 tumor bearing mice. In conclusion, the host defense system of the liver can be modulated to enhance or inhibit colon-derived experimental hepatic metastasis.     

Absorption of inorganic iron- (59Fe2−) in relation to iron stores in pancreatic exocrine insufficiency due to cystic fibrosis

Summary The absorption of⁵⁹Fe from a diagnostic 0.56 mg⁵⁹Fe²⁺ dose was measured by 4-geometry whole body counting and related to the amount of stainable diffuse cytoplasmatic non heme storage iron in the bone marrow macrophages of children with cystic fibrosis. When the storage iron was within the normal range (+/2+) children with cystic fibrosis absorbed 10–38% ( _a±S.D.=21±9.3) of the oral⁵⁹Fe²⁺ dose which is identical with the⁵⁹Fe-absorption in normal children with normal iron stores (9–45:23±8.7%). Depleted iron stores caused an increase of⁵⁹Fe-absorption to 43–95% ( _a±S.D.=62±19) in children with cystic fibrosis and to 45–100% ( _a±S.D.=73±18) in control children. The interruption or continuation of pancreatin maintenance therapy and the simultaneous administration of 1–1.5 g pancreatin did not influence⁵⁹Fe²⁺ absorption in cystic fibrosis.There is no evidence for a pancreatic factor required for or inhibiting inorganic and food iron absorption in human beings. Iron absorption is controlled also in cystic fibrosis chiefly by the amounts of available storage iron. It is therefore not justified to apprehend the development of hemosiderosis in children with cystic fibrosis who are not or not sufficiently treated with pancreatin.Supported in part by research grants of the Deutsche Forschungsgemeinschaft