The use of human serum for studying malformations in whole embryo cultures.
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Abnormalities, Drug-Induced↗
Biomedical subjects
F Beck
Publications and source records attributed to F Beck.
Growth of rat embryos in the serum of alcohol drinkers.
Seven healthy male volunteers who had fasted overnight consumed Scotch whisky (70-85 g absolute alcohol) in a period of 15 minutes after venesection at 9.30 a.m. An hour later a further quantity of blood was collected. Rat embryos (9.5 days of gestation) grown for four hours in 'post-drink' serum (115 mg alcohol/100 ml serum) followed by 44 hours in 'pre-drink' serum were compared to controls cultured in normal human serum for 48 hours. All cultures contained 90% human serum and 10% rat serum. The embryos were examined morphologically and their protein content was measured to assess in vitro growth and differentiation. The results demonstrated the teratogenic and growth-retarding effects of alcohol ingestion. Addition of ethanol (120 mg/100 ml) to the culture medium produced similar results. Culture of 9.5-day rat embryos for 24 hours in 'post-drink' serum (115 mg/100 ml alcohol) containing 10 or 20 micrograms acetaldehyde/ml or in pre-drink serum containing similar amounts of acetaldehyde showed a toxic effect of acetaldehyde only at concentrations of 20 micrograms/ml, in the absence of alcohol.
Abnormalities, Drug-Induced↗
Intracellular electrolyte concentrations in rat sympathetic neurones measured with an electron microprobe.
Intracellular element concentrations were measured in rat sympathetic neurones using energy dispersive electron microprobe analysis. The resting intracellular concentrations of sodium potassium and chloride measured in ganglia maintained for about 90 min in vitro at 25 degrees C were 3, 155 and 25 mmol/kg total tissue wet weight respectively. Recalculated in mmol/l cell water, these values are 5, 196 and 32 respectively. There were no significant differences between the nuclear and cytoplasmic values of these ions. Incubation in either carbachol (180 mumol/l, 4 min) or ouabain (1 mmol/1, 60 min) significantly increased the intracellular sodium and decreased the intracellular potassium concentrations. Neither substance materially altered the intracellular chloride concentration. The data obtained are compared and contrasted to those obtained in mammalian sympathetic neurones using chemical analysis and ion-sensitive microelectrodes.
Animals↗
Intra- and extracellular element concentrations of rat renal papilla in antidiuresis.
The element concentrations in various intra- and extracellular compartments of the tip of the rat renal papilla were determined during antidiuresis using electron microprobe analysis. Urinary concentrations (means +/- SEM) were: urea, 1509 +/- 116; potassium, 268 +/- 32; sodium, 62 +/- 19 mmoles X 1(-1); and osmolality, 2548 +/- 141 mOsm X kg-1. Electrolyte concentrations in the interstitial space were: sodium, 437 +/- 19; chloride, 438 +/- 20; and potassium, 35 +/- 2 mmoles X kg-1 wet wt. The vasa recta plasma exhibited almost identical element concentrations. The values in the papillary collecting duct cells were: sodium, 28 +/- 1; chloride, 76 +/- 3; potassium, 135 +/- 3; and phosphorus, 316 +/- 7 mmoles X kg-1 wet wt. Similar concentrations were observed in the papillary epithelial cells. In interstitial cells potassium and phosphorus concentrations were virtually identical to those of the collecting duct cells, whereas sodium and chloride concentrations were higher by about 30 mmoles X kg-1 wet wt. The element composition of the various papillary cells is, thus, not substantially different from that of proximal tubular cells. This finding demonstrates that cellular accumulation of electrolytes is not the regulatory mechanism by which papillary cells adapt osmotically to their high environmental osmolality and sodium chloride concentration.
Adaptation, Physiological↗
Intracellular electrolyte composition in various experimental models of hypertension: an electron microprobe study.
Changes in the intracellular ionic composition in the three models of hypertension studied are not uniform in the cells of the various organs. The composition differs not only from organ to organ, but even among the various cell types within the same organ. Marked differences--even diametrically opposite changes--in the intracellular Na concentration can be detected in the various models of hypertension studied. Hence, one cannot expect to draw a simple unifying hypothesis from an analysis of the changes in intracellular electrolyte concentrations occurring in hypertension. A more quantitative analysis of the intracellular ionic composition in other cells, particularly in vascular cells, is needed to define the characteristics of the various types of hypertension.
Animals↗
Characterization of exocoelomic fluid protein from rat conceptuses cultured in rat and human sera: a measure of yolk sac activity during organogenesis.
Fluid from the extraembryonic coelom of 11.5-day rat embryos cultured in 100% rat serum, 100% human serum and 90% human serum supplemented with 10% rat serum between days 9.5 and 11.5 postconception were compared using polyacrylamide gel electrophoresis and crossed immunoelectrophoresis. The protein composition of the exocoelomic fluids differed considerably from one another and from each of their respective culture sera. The majority of proteins in the exocoelom were derived from macromolecular transport but some contribution was made from protein synthesis by the conceptus. Eighteen proteins normally found in rat serum were found in the exocoelom of conceptuses cultured in 100% rat serum. Eighteen proteins were found in the exocoelom of rat conceptuses cultured in 100% human serum, of which ten were derived from human serum and eight were proteins normally found in rat serum. Analysis of fluid from conceptuses cultured in 90% human serum supplemented with 10% rat serum showed eleven human serum proteins and ten rat serum proteins. Differences in the composition of both human and rat proteins between the latter two fluids were also evident.
Animals↗
Normal and abnormal appearances of the 35-day ferret fetus studied by freehand (razor blade) sections and gross examination.
This communication reports the sectioning and subsequent examination of 1-mm freehand razor blade sections of the normal 35-day ferret fetus; pregnancy in the ferret lasts for 42 days. Skeletal and other abnormalities obtained after maternal treatment with teratogens, such as trypan blue, mustine hydrochloride, and sodium salicylate are included for comparison with the normal.
Abnormalities, Drug-Induced↗
Incorrect use of yolk sac cavity.
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Animals↗
Effects of glucose on rat embryos in culture.
To elucidate the role of hyperglycaemia in causing fetal malformation, rat embryos have been grown in vitro from the head-fold stage for 48 h in the presence of excess glucose during different parts of the culture period. Culture of rat embryos for 10 or 21 h in 55 mmol/l exogenous glucose produced abnormalities observable after 48 h in culture. When embryos cultured for 10 or 21 h in excess glucose were observed by scanning electron microscopy at the end of the glucose treatment, abnormalities could be observed which may indicate how later malformations are formed. Thus it is possible that a relatively brief hyperglycaemic episode at a critical stage of embryogenesis may endanger the fetus of a diabetic mother.
Abnormalities, Drug-Induced↗
Effect of aldosterone on the human erythrocyte sodium--potassium pump in vitro.
1. The effects of aldosterone in vitro on the Na+, K+-dependent ATPase activity of isolated human erythrocyte membranes and on rubidium (86Rb) uptake and [3H]ouabain binding of intact erythrocytes were studied. 2. ATPase activity was nearly doubled (0.061 +/- 0.006 to 0.110 +/- 0.01 mumol of Pi h-1 mg-1 of protein) by the addition of a physiological concentration of aldosterone (2.7 X 10(-10) mol/l). Higher concentrations had no greater effect. 3. Aldosterone had no significant effect on 86Rb uptake or [3H]ouabain binding. 4. Erythrocytes contain aldosterone at concentrations similar to that in plasma. The effect of aldosterone on ATPase is probably maximal.
Adult↗
Reduced sodium-potassium dependent ATPase and its possible role in the development of hypertension in spontaneously hypertensive rats.
Ouabain-sensitive Na+-K+-ATPase activity in red cell membranes, kidney cortical tissue, myocardium and adrenal glomerulosa tissue was examined in SHR and WKY rats at 6, 9, and 12 weeks of age. Red cell membrane enzyme activity was decreased (p less than 0.001) at 9 and 12 weeks of age in SHR. This activity was negatively correlated (r = -0.69, p less than .005) with blood pressure at 9 and 12 weeks. Kidney cortical enzyme activity was also decreased (p less than 0.001) in the SHR at 9 and 12 weeks of age. This decreased kidney enzyme activity was also inversely related to 9 and 12 week blood pressures (r = -0.71, p less than 0.001), urinary Na excretion (r = -0.62, p less than .005), and urinary Ca and K excretion. Myocardial enzyme activity was not decreased until 12 weeks in the SHR, and adrenal glomerulosa activity was not different in the SHR and WKY at any of the three ages that this enzyme was measured. Of the tissues examined decreased Na+-K+-ATPase activity in the erythrocyte membrane and in kidney cortical tissue appears to coincide best with the development of hypertension in the SHR. This study lends further support to the concept that alterations in membrane cation transport may be an important factor in the development of high blood pressure in SHR.
Adrenal Glands↗
Arterial wall renin.
Aortic homogenates contain proteolytic enzymes which will release angiotensin I (AI) from renin substrate. Some of these are active at low pH and are probably unrelated to renin. Renin-like activity in the rat measured at the optimum pH of 6.5 is altered in parallel with plasma renin in a wide variety of situations. The two diverge only in non-steady state situations. Studies have therefore been carried out after bilateral nephrectomy and after the injection of renal renin into nephrectomized rats. In each case aortic renin-like activity was cleared much more slowly than plasma renin, and the blood pressure change was related to aortic renin-like activity rather than to plasma renin. The blood pressure response to the converting enzyme inhibitor teprotide was also related to the former rather than the latter. Immunofluorescent studies of the aorta and the intrasplenic arteries from rats injected with mouse renin showed that renin was taken up predominantly into the media and persisted at this site. Thus, the uptake of renal renin from plasma by both large and small arteries is probably an important step in the physiology of the renin-angiotensin system and mediates renin-induced changes in peripheral resistance and blood pressure. However, we have no evidence for the hypothesis that selective accumulation of renin in the resistance vessel walls causes hypertension when circulating levels of renin are normal.
Animals↗
Sodium and potassium concentrations of renal cortical cells two animal models of primary arterial hypertension.
Electron microprobe analysis was used to determine cellular concentrations of potassium and sodium in renal cortical cells of hypertensive rats of the Milan strain (MHS) and spontaneously hypertensive rats of the stroke prone strain (SHRSP) and their respective controls. Potassium concentrations in proximal and distal tubular cells were similar in both strains of hypertensive rats compared with their normotensive controls. In MHS rats proximal tubular cell sodium concentration was lower than in controls by 3.1 mmol/kg ww, whereas in both proximal and distal tubular cells of SHRSP sodium concentrations were higher than in controls by 5.3 and 4.3 mmol/kg ww, respectively. These results indicate that changes in the transport characteristics of the renal tubular epithelium are a feature of both models of hypertension.
Animals↗
Morphological, total nucleic acid and total protein analyses of rat embryos cultured in supplemented and unsupplemented human serum.
Culture of explanted 9 1/2 day rat conceptuses for 48 hours (i.e. until 11 1/2 days) in 100% human serum supplemented only with glucose does not result in growth and development comparable to those conceptuses cultured for the same period in 100% rat serum. Significant differences were observed in the yolk sac diameter, crown-rump length, morphological score and total embryonic nucleic acid content. No difference was observed in the total embryonic protein content. A large proportion of the embryos cultured in glucose-supplemented human serum (100%) exhibited abnormalities of the neural tube. Supplementation of the human serum with glucose and 10% rat serum resulted in embryos showing higher indices of embryonic growth and differentiation (in all criteria of assessment) than those cultured in pure rat serum; no abnormalities were observed. The significance of better growth and differentiation of the conceptuses in human serum supplemented with glucose and 10% rat serum is discussed.
Animals↗
Maturation of the ileal epithelium in the young rat.
Normal maturation in the rat ileal epithelium (ileal 'closure') occurs between 18 and 21 days, probably by a process of cellular replacement. A single dose of cortisone acetate induced a precocious ileal 'closure' within 6 days. The morphological changes observed were similar to those seen during normal 'closure'. Morphometric measurements made of ileal villi during both normal and induced 'closure' suggest that there was a gradual change in the type of cells produced by the crypt. This was perhaps related to the duration of exposure of the cryptal cells to the 'closure' stimulus. Ten days after cortisone acetate treatment vacuolated cells were again seen on the villus in some animals. The variation in their distribution on the villus was perhaps due to differing rates at which the villi had recommenced pinocytosis because of the physiological state of the animal. A single dose of corticosterone failed to promote any morphological changes indicative of ileal 'closure' when observed at the electron microscope level.
Animals↗
Growth of 9.5-day rat embryos in human serum.
Rat embryos were explanted at 9.5 days and cultured for 48 h in human serum supplemented with glucose. The embryos were anaemic and frequently malformed. The haemoglobin and DNA content of these embryos was less than those grown in pure rat serum. Addition of 10% rat serum improved the embryonic growth as well as the haemoglobin and DNA content. This suggests that rat serum provides essential growth factors required by the embryos which are not present in human serum.
Anemia↗
Maturation in the ferret ileal epithelium and the effect of cortisone acetate.
Normal maturation ('closure') in the ferret ileal epithelium occurs, between 35 and 39 days after birth, by a process of cellular replacement. The administration of cortisone acetate at doses up to 1.2 mg/g body weight for 5 days failed to promote any morphological changes in the ileal epithelium in this non-rodent species, and the results suggest that the maturing effect of cortisone acetate in the rat might be peculiar to the rodents.
Aging↗
Culture of 91/2-day rat embryos in human serum supplemented and unsupplemented with rat serum.
We attempted to grow explanted 91/2-day rat embryos in vitro in human serum but were unable to demonstrate normal growth as has been previously described. The embryos were consistently smaller, anaemic, and frequently deformed. These abnormalities were overcome by supplementing the human serum with 10% rat serum. The latter possibly provides yet unidentified essential growth factors required by rat embryos that are not present in human serum.
Animals↗