Effect of early postnatal long-term fasting on the development of peptide hydrolysis in chicks.

1. Early development of peptide hydrolysis in the digestive tract was investigated in experiments with fasted and fed ad lib. chicks during the first decade of postnatal period. 2. Pancreatic carboxypeptidase A (CPA) activity was maximal at the moment of hatch. On the second day CPA activity considerably diminished in starved and fed animal groups; further starvation (3-4 days) led to the significant increase of CPA total and specific activity, whereas the amount of enzyme in pancreas of fed chicks was rather low. 3. Aminopeptidase (AP) activity of the small intestinal surface was less sensitive to starvation. The increase of activity in all intestinal parts was observed only on the 4th day of fasting. The most sensitive to starvation were dipeptidases. Changes in their activity (2-fold increase) were detected after 24 hr of starvation. 4. The formation of specific physiological proximo-distal gradient of intestinal exopeptidase activities began only after the moment of the first feeding. 5. This gives evidence that the development of peptide hydrolysis depends not only on the age of the animal but also on the normal physiological beginning of the process of exogenous nutrition.     

Hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and cholesterol 7 alpha-hydroxylase activity in neonatal guinea pig.

Optimal assay conditions for hepatic HMG-CoA reducatase activity and cholesterol 7 alpha-hydroxylase activity in the guinea pig were determined. These two enzyme activities were studied in the liver of newborn guinea pigs during the first three postnatal weeks. Hepatic HMG-CoA reductase activity of neonatal guinea pigs was similar to that of adult animals. However, cholesterol 7 alpha-hydroxylase activity of newborns was about one-third of that in adult guinea pigs. This finding suggests that the system for bile acid synthesis in the neonatal guinea pigs is underdeveloped.     

Cytochemical analysis of single villus peptidase activities in pig intestine during neonatal development

The present work uses a new technique of whole tissue cytochemistry and automated scanning to obtain measurements of peptidase activity and surface structure in intact villi microdissected from the jejunum of newborn and 28-day-old pigs. Intact villi from 28-day-old pigs are shown by this method to contain 30% more aminopeptidase N and 400% more dipeptidylpeptidase IV activity than is found on villi taken from newborn pig intestine. Villi taken from 28-day-old pig intestine are also half as long and twice as wide at their base as those taken from newborn animals. These changes in shape take place without significantly affecting the total surface area of the villus. Increases in peptidase activities occurring during postnatal development can be further subdivided into those dependent upon induced changes in enterocyte biochemistry and those dependent upon the changing geometry of villi. Over 90% of the total increase in peptidase activities occurring during neonatal development was shown, by this analysis, to involve enterocyte reprogramming of enzyme production. The present method of whole tissue cytochemistry appears to combine successfully the ability to measure peptidase activities at the cellular level in undisrupted tissue with a facility to relate these results to the overall shape of a single villus. These and more general applications of the method now provide new ways to analyse a variety of changes taking place in intestinal structure and function in a quantitative manner.     

Placental transfer of cholesterol-4-14C into rabbit and guinea pig fetus

A tracer dose of cholesterol-4-(14)C was given daily in the diet of six pregnant guinea pigs to establish an isotopic steady state. At the time of parturition, maternal and fetal blood and fetal tissues were collected and analyzed for cholesterol content and cholesterol specific activity. A comparison of these specific activities in neonatal and maternal serum indicated that about 22% of the fetal serum cholesterol was transferred from maternal blood. In the newborn, tissues generally had the same cholesterol specific activity as serum. Brain tissue was an exception in having a specific activity only 8.4% of that of serum. Dietary cholesterol did not increase serum cholesterol levels in the newborn but did increase the percentage of fetal cholesterol derived from the maternal circulation. The rapid transfer of cholesterol-4-(14)C across the placenta was indicated by the appearance of this isotope in the newborn 2 days after its administration to pregnant rabbits. A considerable amount of the cholesterol content of newborn guinea pigs and rabbits originated from the maternal blood.     

Adénylate-cyclase/Lipase: Induction des récepteurs hormonaux

An adenylate cyclase activity (AC) was found in guinea pig brown adipose tissue (BAT), since the tissue's apparition. This enzymatic activity increased during the development and showed high values at the end of gestation. An increase of AC units per cell was observed, in addition to the cell multiplication. A norepinephrine stimulation of AC activity was observed at the end of gestation: this regulating action disappeared in the first days of extrauterine life. Neither glucagon nor ACTH had any regulating role upon AC activity during fetal and newborn life. The basal lipolytic activity which was observed in BAT of fetuses (61rst day) and neonate dramatically decreased around the 15th day. A potent lipolysis activation by norepinephrine was observed, but only after birth. The correlation observed between these enzymatic activities in presence of norepinephrine seems to indicate that the AC/lipase system was involved in the neonatal thermogenesis of guinea pigs.     

Naloxone does not alter the "regulated" decrease in core temperature during hypoxemia in guinea pigs

In newborns andadults of a number of species, exposure to acute hypoxemia produces a"regulated" decrease in core temperature, the mechanism of whichis unknown. The present experiments were carried out in chronicallyinstrumented newborn (5-10 days of age;n = 59) and older (25-30 days ofage; n = 61) guinea pigs to test thehypothesis that the endogenous opioids mediate this regulated decreasein core temperature. During an experiment, core temperature, oxygenconsumption, and selected ambient temperature were measured in athermocline (linear temperature gradient of 10-40°C) during acontrol period of normoxemia, an experimental period of normoxemia orhypoxemia (inspired oxygen fraction 0.10), and during a recovery periodof normoxemia following an intraperitoneal injection of naloxonehydrochloride (a nonspecific opioid antagonist; 1, 2, or 4 mg/kg) orvehicle. Naloxone did not significantly alter basal core temperature orthe core temperature response to acute hypoxemia in newborn or olderguinea pigs. Naloxone did, however, decrease basal oxygen consumptionin newborn and older guinea pigs and altered the thermoregulatoryeffector mechanism used to decrease core temperature during hypoxemiain the newborn guinea pigs. Our data do not support the hypothesis thatthe endogenous opioids mediate the regulated decrease in coretemperature that occurs in newborn and older guinea pigs duringexposure to acute hypoxemia.

     

Long-Term Responses to Loss of Renal Mass: Glomerular Changes: Evidence of Induction of New Nephrons in Immature Kidneys Undergoing Hypertrophy

Unilateral nephrectomy performed during the first days or weeks of life may or may not induce the development of new nephrons in the remaining kidney. Such an increase has been reported to occur, as discussed in this review, in newborn rats and mice, but not in guinea pigs. These observations are consistent with other data suggesting different patterns of compensatory adaptation in young and in old animals.     

Postnatal development of transport function in the pig intestine

1. In the newborn pig it appears that only prenatally produced enterocytes are capable of absorbing large amounts of protein. 2. The ability of the small intestine to transport sodium, lysine, lysine containing dipeptides and glucose declines markedly during the first week of post natal life. 3. Dexamethosone causes a doubling of the sodium dependent portion of alanine uptake. 4. EGF given between days three and six of postnatal life increases sucrase and maltase activity in the distal region of the small intestine. 5. Weaning induced problems are probably not due to direct inhibition of transport properties.     

Maturation of the activity of cerebellar Purkinje cells and the lift reaction

In experiments on guinea pigs (from newborn to adults), studies have been made on the extensor, support and lift reactions, as well as on the activity of cerebellar Purkinje cells in the same animals. First signs of immature lift, extensor and support reactions were observed already 12th after birth. At this period, mean discharge frequency in Purkinje cells was significantly lower than in the adult animals, reaching 11.5 +/- 1.2 imp/s for simple spikes and 0.45 +/- 0.05 imp/s for complex ones. Complete maturation of lift, extensor and support reactions takes place to the beginning of the 2nd week (8-9 days) of postnatal life. Within this period, significant changes in the activity of Purkinje cells were observed: mean discharge frequency of simple and complex spikes increased correspondingly up to 17.9 +/- 2.3 and 1.48 +/- 0.25 imp/s. At the same time, the mean discharge frequency in Purkinje cells, the average duration of inhibition pause, and the response latency became more stable.     

Effect of cyclopiazonic acid on delayed hypersensitivity to Mycobacterium tuberculosis,complement activity,serum enzymes,and bilirubin in guinea pigs

Cyclopiazonic acid (CPA) was given daily to groups of guinea pigs at doses of 0.00625, 0.0125, 0.025, 0.05, 0.1, 0.2, 0.4, 0.8, 1.6, and 1.95 mg/day for 30 days. All guinea pigs were sensitized and survivors were skin tested twenty-five days later with Mycobacterium tuberculosis. Mortalities occurred only in the two greatest dose groups. Signs of disease included anorexia, roughened hair coat, diarrhea and incoordination. The major histopathologic changes occurring in these two groups included hepatocellular vacuolar degeneration and necrosis of the gastric mucosa with infiltration of neutrophils in the deep gastric mucosa. CPA did not affect cutaneous hypersensitivity to M. tuberculosis, complement activity, serum glycocholic acid concentrations or weight gains. There were increases in aspartate aminotransferase, alanine aminotransferase, and sorbitol dehydrogenase concentrations in the serum of guinea pigs in the two greater dose groups, but no changes were found in serum concentrations of SAP. There was a slight increase in the serum bilirubin concentrations in the greater dose groups.Note endorsements are herein implied.     

Species comparison of adenosine A1 receptors in isolated mammalian atrial and ventricular myocardium

Various species have been used as models to study the effects of adenosine (ADO) on atrial and ventricular myocardium, but few direct tissue comparisons between species have been made. This study further characterizes adenosine A(1) receptor binding, adenylate cyclase activity and direct and indirect A(1) receptor-mediated functional activity in atrial and ventricular tissue from Sprague-Dawley rats and Hartley guinea pigs. Rat right atria (RA) were found to be significantly more sensitive to cyclopentyladenosine (CPA), while guinea pig left atria (LA) were more sensitive to CPA. After the addition of isoproterenol (ISO), the reduction of CPA response in rat RA was significantly greater than in guinea pig; however, after ISO treatment, the guinea pig LA was more sensitive to CPA than the rat. Adenylate cyclase inhibition by CPA was significantly greater in atria and ventricles obtained from guinea pig than rat. In competition binding experiments, guinea pig RA had significantly more high affinity sites than rat, but the K(i)s were not significantly different. There were no significant differences between guinea pig LA and rat LA. Guinea pig ventricular tissue had fewer high affinity sites than rat without any differences in their K(i) values. In antagonist saturation experiments, the density and affinity of A(1) receptors in guinea pig cardiac membranes were significantly greater than in rat. Our results indicate definite species differences as well as tissue differences between rat and guinea pig. These differences must be considered when interpreting studies using rat and guinea pig tissue as models for cardiac function.     

Cellular energy metabolism during fetal development. IV. Fatty acid activation, acyl transfer and fatty acid oxidation during development of the chick and rat

We have investigated developmental profiles of ATP-dependent palmityl-CoA synthetase, acetyl-CoA synthetase, palmitylcarnitine transferase, and fatty acid oxidation in heart and liver of developing chicks and rats. Palmityl-CoA synthetase activity of rat liver and heart homogenates increased 6- to 10-fold during the first postnatal week. Chick embryo heart activity peaked between 13 and 16 days of development. The activity of embryonic chick livers was bimodal with highest activity seen at 7 and 16 days of development. Posthatching values were approximately 50–75% of the peak embryonic levels. Acetyl-CoA synthetase activity of rat liver and heart homogenates was low but also showed developmental increases following birth. Acetyl-CoA synthetase activity of chick embryonic hearts was greatest at 16 days of development. Palmitylcarnitine transferase activity of rat liver and heart homogenates showed a striking increase during the first week of life. Chick heart activity was similar to that observed for palmityl-CoA synthetase with a peak between 13 and 16 days of embryonic development. Coincident with the postnatal rise in fatty acid activation and palmitylcarnitine transferase activity in developing rats, the oxidation of palmityl-CoA plus carnitine and of palmitylcarnitine increased from barely measurable levels at birth to adult levels by 30 days of age. The increases that we observe probably relate to changes in the specific activity of the enzymes as well as to an increase in the absolute number of mitochondria during development.     

Development of gastric acid secretion in pigs from birth to thirty six days of age: the response to pentagastrin

The development of the gastric acid secretory response to pentagastrin was studied using 56 Large White x Landrace pigs, 0-36 days of age, 1.1-13.3 kg body-weight, obtained from 12 litters. Gastric acid secretory capacity was measured using a gastric perfusion technique and intravenous infusion of pentagastrin at dose rates of 2, 4 and 8 micrograms/h per kg. Significant positive linear correlations were found between stomach weight and age, and between stomach weight and body-weight during the 36 day period. The stomach weight to body-weight ratio increased for the first 3 days of age and then decreased during the following 33 days. Basal acid secretion was detected in all unsuckled pigs (n = 9), 2- to 8-h old. Maximal acid outputs in response to pentagastrin in these pigs were 0.16 +/- 0.02 mmol/kg body-weight and 0.034 +/- 0.001 mmol/g stomach weight. For the 56 pigs, significant linear correlations were found between maximal acid output and age, maximal acid output and body-weight, and maximal acid output and stomach weight. There was a significant linear increase in maximal acid output per unit stomach weight during the first 7 days of age, but during the subsequent 29 days the pattern of increase in gastric secretory capacity was slower and curvilinear. In the oldest nine pigs, 24-36 days of age, maximal acid outputs were 0.974 +/- 0.058 mmol/kg body-weight and 0.234 +/- 0.016 mmol/g stomach weight which represents a six to seven-fold increase compared with those determined in pigs at birth. Comparison of gastric acid secretory capacity determined under anaesthesia with that in conscious pigs showed that anaesthesia appeared to suppress basal output but had no effect on pentagastrin stimulated output. Comparison of response to histalog (betazole HCl) and pentagastrin indicated that newborn pigs were more sensitive to histalog but in pigs 9-38 days of age, there were no significant differences in responsiveness to the two secretagogues. These results show that gastric sensitivity to pentagastrin increases rapidly in the first week of life, that the stomach of the newborn pig is more sensitive to histalog than pentagastrin and that studies of the effect of pentagastrin on acid secretion, done under anaesthesia, are comparable to those in the conscious pig.     

Protective effects of antibody against intestinal invasion by Escherichia coli

Seven Escherichia coli isolates from newborn calves with diarrhea were examined for enteropathogenic properties. One isolate penetrated into HeLa cells, four produced enterotoxin(s) and the remaining two possessed neither of these properties. Penetration of E. coli into HeLa cells was inhibited by antibody in bovine colostrum and in bovine and rabbit immune sera. The effective antibodies appeared to be mostly of the IgM class. The invasion by E. coli isolates was also examined by inoculation of the bacteria into the small intestine of E. coli-immunized and non-immunized guinea pigs. The isolate which penetrated into HeLa cells could penetrate the intestinal mucosa to be disseminated into various organs of non-immunized guinea pigs, but not of immunized guinea pigs, whereas no other isolates showed such pathogenicity in vivo. The inhibition of the invasion was observed when non-immunized guinea pigs were inoculated with the bacteria together with colostral or serum antibody. The results show the importance of antibody in the local defense mechanism against E. coli invasion.     

Effect of starvation and a protein diet on the amino acid metabolism enzyme activities of the organs of domestic fowl hatchlings

The activities of alanine and aspartate transaminases, adenylate deaminase, glutamine synthetase and glutamate and xanthine dehydrogenases have been measured in liver, yolk sac membrane, intestine and breast and leg muscle of domestic fowl hatchlings receiving for 3 or 5 days either a standard diet or hard boiled eggwhite as well as in 3 or 5 days starved animals. The patterns of activation of amino acid metabolism enzymes were fully comparable in protein-fed and starved groups with respect to fed controls; the differences with respect to the latter became more marked in 5- than in 3-days old chicks. In 5-days old chicks intestine alanine transaminase activity increased in parallel to that of liver in protein-fed animals but not in those starved, in agreement with an enhanced alanine transfer between both organs under this situation. Both, starvation and protein-feeding, induced a general decrease in the amino acid metabolizing ability of muscle. Glutamine (but not alanine) synthetizing capabilities were enhanced.     

A possible mechanism for the changes in hepatic and intestinal alkaline phosphatase activities in bile-duct-ligated rats or guinea pigs

The effects of bile-duct ligation on hepatic and intestinal (jejunum) alkaline phosphatase activities were studied using rats and guinea pigs. In ligated rats, the enzyme activity was increased 4.1-fold in the liver after 24 h and 2.8-fold in the intestine after 12 h. In guinea pigs, the hepatic and intestinal enzyme activities were increased 2.3-fold and 1.5-fold after 100 and 24 h, respectively. The intestinal activity was induced sooner after ligation than hepatic activity. The induction of alkaline phosphatase was inhibited by prior treatment of animals with amanitin, an inhibitor of RNA polymerase activity. This result indicates that the induction is associated with de novo enzyme synthesis. The content of cyclic AMP in liver and intestine increased immediately after ligation. The increase in alkaline phosphatase activities was also inhibited by pretreatment with chlorpromazine, an inhibitor of adenylate cyclase activity. Hence, cellular cyclic AMP may be implicated in playing a role in the induction of alkaline phosphatase by bile-duct ligation.     

Differences in activities of antioxidant superoxide dismutase, glutathione peroxidase and prooxidant xanthine oxidoreductase/xanthine oxidase in the normal corneal epithelium of various mammals

Under normal conditions, antioxidants at the corneal surface are balanced with the production of reactive oxygen species without any toxic effects. Danger from oxidative stress appears when natural antioxidants are overwhelmed leading to antioxidant/prooxidant imbalance. The aim of the present study was to examine the activities of enzymes contributing to the antioxidant/prooxidant balance in normal corneal epithelium of various mammals. The enzyme activities of antioxidant superoxide dismutase and glutathione peroxidase, as well as prooxidant xanthine oxidoreductase/xanthine oxidase were examined using biochemical methods. Results show that superoxide dismutase activity is high in rabbits and guinea pigs, whereas in pigs the activity is low and in cows it is nearly absent. In contrast, glutathione peroxidase activity is high in cows, pigs and rabbits, whereas in guinea pigs the activity is low. As far as prooxidant enzymes are concerned, elevated xanthine oxidoreductase/xanthine oxidase activities were found in rabbits, lower activities in guinea pigs, very low activity in cows and no activity in pigs. In conclusion, the above results demonstrate inter-species variations in activities of enzymes participating in antioxidant/prooxidant balance in the corneal epithelium. It is suggested that the levels of antioxidant and prooxidant enzymes studied in the corneal epithelium might be associated with the diurnal or nocturnal activity of animals. UV rays decompose hydrogen peroxide to damaging hydroxyl radicals and perhaps for this reason large animals with diurnal activity (cow, pig) require more effective peroxide removal (high glutathione peroxidase activity) together with the suppression of peroxide production (low superoxide dismutase activity, low xanthine oxidoreductase activity).     

Developmental changes in endothelium-derived vasorelaxant factors in cerebral circulation

Endothelium-derived prostanoids are predominant vasorelaxant factors in the cerebral circulation of newborn pigs in vivo, whereas in older pigs nitric oxide (NO)-mediated responses also contribute to the regulation of cerebral vascular tone. We compared the expression and activities of NO synthase and cyclooxygenase in the cerebral microcirculation of newborn and adult pigs. In adult animals, expression and activity of endothelial NO synthase in cerebral microvessels and in cultured cerebral endothelial cells is two- to threefold higher than in newborn pigs; acetylcholine and bradykinin cause a greater increase in NO production in adult pigs. Expression and activity of cyclooxygenase in cerebral microvascular endothelial cells is similar in newborn and adult pigs; acetylcholine and bradykinin stimulated dilator prostanoid production to the same degree in both age groups. Endothelial prostanoid synthesis in cerebral microvessels and cultured endothelial cells was inhibited 30-70% by NS-398, reflecting a large contribution of COX-2 in both newborn and adult animals. These data indicate that in the cerebral circulation of pigs, NO synthase is age-dependently upregulated, whereas endothelial cyclooxygenase is not altered during postnatal development.     

Proteinase,Lipase and Amylase Activities in the Small Intestine of Pigs Suffering from Colienterotoxaemia

The activities of proteinases, lipases, amylases and the activities of proteinase inhibitors, as well as the numbers of Escherichia coli in the contents from the small intestine were examined for pigs suffering from colienterotoxaemia and for healthy pigs. Enzyme activities were determined using an agar diffusion test. Compared with healthy animals the activities of proteinases and amylases in diseased animals were reduced while lipases showed increased activity. In pathologically changed contents showing large numbers of E. coli, proteinases could not be demonstrated; however, proteinase inhibitors were found in these contents. In healthy animals, proteinase inhibitors were not demonstrated in ingesta-con-taining contents. In diseased animals, E. coli were found in large numbers in all parts of the small intestine. In healthy animals, E. coli was demonstrated especially in the posterior part of the small intestine and regularly in small numbers. The possible influence of digestive enzymes, especially proteinases and their inhibitors, on enterotoxins from E. coli is discussed.     

Glucose-6-phosphatase activity in mouse small intestine during postnatal development.

The postnatal development of phosphohydrolase activity of glucose-6-phosphatase has been examined in the different parts of the small intestine of the mouse and compared with that of trehalase and glucoamylase. After birth, glucose-6-phosphatase is present in all parts of the small intestine and the activity is already two to three times the adult values. In the course of the first 2 weeks, the activity increases and the different parts of the small intestine show distinct patterns. One day after birth, glucoamylase activity is present in all segments, while trehalase activity is confined to the first intestinal third. During the first 2 weeks, these activities remain rather stable. In the course of the third week, when trehalase and glucoamylase are increasing, glucose-6-phosphatase activity is declining toward adult values.