28th Day Of Age Research Articles

Effects of ascorbic acid on the vasoactive intestinal peptide synthesis in the ileum submucous plexus of normal rats

The aging process is a deteriorating process that attacks the gastrointestinal tract, causing changes in the number and size of neurons from the enteric nervous system. The activity of free radicals on enteric neurons is helped by the significant reduction of antioxidants. Evaluate the effect of the ascorbic acid supplementation on the neurons that produce the vasoactive intestinal peptide (VIP) in the submucous plexus of the ileum of normal rats for a period of 120 days. Fifteen rats were divided in three groups: untreated control with 90 days, untreated control with 210 days and ascorbic acid-treated rats with 210 days. Ascorbic acid was given for 16 weeks from the 90th day of age by adding it to drinking water (1 g/L prepared fresh each day). The ileums were processed according to the immunohistochemistry technique for whole-mount preparation in order to detect the presence of VIP immunoreactive in the cellular bodies and nervous fibers in the neurons of the submucous plexus. We have verified their immunoreactivity and measured the cellular profile of 80 cellular bodies of VIP-ergic neurons from each studied group. The ascorbic acid supplementation did not alter physiological parameters such as water intake and food consumption of the three studied groups. We observed a significant increase of the cellular profile of VIP-ergic neurons in untreated control with 210 days when compared to untreated control with 90 days. The cellular profile of VIP-ergic neurons in ascorbic acid-treated rats with 210 days was bigger than those observed in others groups. The ascorbic acid had a neurotrophic effect on VIP-ergic neurons on the ileum after period 120 days of supplementation.

Dependence on oxygen at the 28th day of age in newborns weighting 1500 g: Incidence and predisposing factors

Dependence on oxygen at the 28th day of age in newborns weighting 1500 g: Incidence and predisposing factors

Comparative effects of different legume protein sources in weaned piglets: nutrient digestibility, intestinal morphology and digestive enzymes

A total of 30 male piglets (Duroc×Landrace) weaned at the 28th day of age were used to compare the nutrient digestibility of legume seeds and their impact on digestive enzyme activities and morphology of intestinal mucosa sampled at slaughter. Five groups of six piglets were fed a control (C) diet based on casein or a diet in which 50% of crude protein (CP) were supplied by soya bean meal (SBM), pea (P), faba beans (FB), or blue lupin ( Lupinus angustifolius) (L), respectively. Total tract (TTAD) and ileal (IAD) apparent digestibilities were measured on the same piglets between the 32nd–39th day and 53rd–60th day of age, respectively. TTAD and IAD of DM, CP and energy were lower in legume-containing diets as compared to the control diet. Among the legume seeds, the lupin-based diet had the lowest TTAD and IAD of DM and energy, but no significant differences were found for the IAD of CP. The height of intestinal villi was the highest with SBM and the lowest with FB only in the duodenum. Crypt depth increased by 50% in the piglets fed the P and FB diets, as compared to controls. Pancreas weight was higher and the duodenal activities of most intestinal enzymes examined were lower with the legume-containing diets.

Chronic postnatal administration of methylmalonic acid provokes a decrease of myelin content and ganglioside N-acetylneuraminic acid concentration in cerebrum of young rats

Levels of methylmalonic acid (MMA) comparable to those of human methylmalonic acidemia were achieved in blood (2-2.5 mmol/l) and brain (1.35 umol/g) of rats by administering buffered MMA, pH 7.4, subcutaneously twice a day from the 5th to the 28th day of life. MMA doses ranged from 0.76 to 1.67 umol/g as a function of animal age. Control rats were treated with saline in the same volumes. The animals were sacrificed by decapitation on the 28th day of age. Blood was taken and the brain was rapidly removed. Medulla, pons, the olfactory lobes and cerebellum were discarded and the rest of the brain ("cerebrum") was isolated. Body and "cerebrum" weight were measured, as well as the cholesterol and triglyceride concentrations in blood and the content of myelin, total lipids, and the concentrations of the lipid fractions (cholesterol, glycerolipids, phospholipids and ganglioside N-acetylneuraminic acid (ganglioside-NANA)) in the "cerebrum". Chronic MMA administration had no effect on body or "cerebrum" weight, suggesting that the metabolites per se neither affect the appetite of the rats nor cause malnutrition. In contrast, MMA caused a significant reduction of plasma triglycerides, but not of plasma cholesterol levels. A significant diminution of myelin content and of ganglioside-NANA concentration was also observed in the "cerebrum". We propose that the reduction of myelin content and ganglioside-NANA caused by MMA may be related to the delayed myelination/cerebral atrophy and neurological dysfunction found in methylmalonic acidemic children.

Effects of ACTH applications during pregnancy on the female offsprings' endocrine status and behavior in guinea pigs

The present study was conducted to elucidate the effects of ACTH application during pregnancy on the female offsprings' endocrine status and behavior in a precocial animal, the guinea pig. We, therefore, studied daughters whose mothers were treated three times with either ACTH-depot or placebo during the sensitive phase of central nervous sexual differentiation of the foetus. From their 41st through their 80th day of age the spontaneous behavior of the daughters was recorded in their home cages, and concentrations of serum cortisol were determined from the 20th through the 90th day of age every 14 days. Daughters whose mothers had received ACTH displayed significantly lower amounts of a defensive aggressive behavior pattern, the urine spray, and they had higher cortisol concentrations than daughters whose mothers had received placebo. Thus, the number of species that shows consequences of an ACTH treatment during pregnancy for the offsprings' endocrine status and behavior is extended to include a precocial animal, the guinea pig.

Morphologic and quantitative study of the myenteric neurons of the jejunum of malnourished rats (Rattus norvegicus).

We studied the effects of maternal proteic desnutrition on the neurons of the myenteric plexus of the jejunum of rats from Rattus norvegicus species. It was used litters of female rats which received diet with normal proteic level during gestation and lactation (group NN), normal diet during gestation and hypoproteic diet during lactation (group ND); hypoproteic diet during gestation and normal diet during lactation (group DN); hypoproteic diet during both gestation and lactation (group DD). After weaning all the animals received diet of normal proteic level until the 60th day of age, when they were killed. The jejunum of the animals was subjected to whole-mount preparations stained by the method of Giemsa and used for the morphologic and quantitative analyses of the neurons of the myenteric plexus. We verified that maternal proteic malnutrition does not cause decrease on the number of myenteric neurons per unit area of jejunum in rats, but elicits mechanisms which assure that, when the animal again receives normal proteic level diet (22%) there occurs storage of proteic material on the cytoplasm of the neurons, thus rendering them larger and strongly basophylic.

Proline administration decreases Na+,K+-ATPase activity in the synaptic plasma membrane from cerebral cortex of rats.

Buffered proline was injected subcutaneously into rats twice a day at 8 h intervals from the 6th to the 28th day of age. Control rats received saline in the same volumes. The animals were weighed and killed by decapitation 12 h after the last injection. Cerebral cortex was used for the determination of Na+,K+-ATPase and Mg2+-ATPase activities. Body, whole brain and cortical weights were similar in the two groups. Na+,K+-ATPase activity was significantly reduced (by 20%) in membranes from the proline-treated group compared to the controls, whereas Mg2+-ATPase activity was not affected by proline. In another set of experiments, synaptic plasma membranes were prepared from cerebral cortex of 29-day-old rats and incubated with proline at final concentrations ranging from 0.1 to 2.0 mM. Na+,K+-ATPase activity, but not Mg2+-ATPase activity, was inhibited by 20-30%. Since proline concentrations in plasma of chronically treated rats and of type 11 hyperprolinemic children are of the same order of magnitude as those tested in vitro, the results suggest that reduction of Na+,K+-ATPase activity may contribute to the neurological dysfunction found in some patients affected by type II hyperprolinemia.

Developmental study of hepatic glutamine synthetase in a mouse model of congenital hyperammonemia

The development of hepatic glutamine synthetase (GS; EC 6.3.1.2) activity and expression was studied in 1 to 112 day old sparse-fur (spf) mutant mice, with X-linked ornithine transcarbamylase (OTC, EC 2.1.3.3.) deficiency. The spf/Y mutant mice were found to have a smaller body weight (p < 0.01) yet possessed a larger liver (p < 0.01-0.05) in comparison to normal male mice (+/Y). The neonatal hepatic GS activity was retarded in the spf/Y mice (p < 0.01) but reached normal values by the 28th day of age, after which it increased as compared to the control CD-I mice (p < 0.01). The spf GS activity remained constant from 28 to 56 days, whereas the CD-I GS activity decreased. A further significant increase in the spf GS activity was observed from 56 day to 112 day indicating its adaptation. The decrease of GS mRNA in the spf/Y mice from 28 to 112 days of age (3.72 +/- 0.25 vs 1.68 +/- 0.32, p < 0.01) suggests translational and post-translational modifications in the regulation of GS activity. The changes in the activity and expression patterns of GS could be due to an effect of the OTC mutation on the hepatic ammonia metabolism. This may be indicative of the adaptational processes in the spf mutant mice, which may play a specific role in this animal model to help it to survive with its hyperammonemia.

Effects of cyclosporin treatment on prolactin pulsatility in chronic hyperprolactinemic male rats

Neuroendocrine effects of cyclosporin (CsA) have been demonstrated, but the mechanisms involved are poorly understood. This work was designed to analyse (i) if chronic CsA administration could influence the episodic secretion of prolactin in adult male rats; and (ii) the effects of the chronic administration of the drug, in adult animals with elevated plasma prolactin levels. Male rats were implanted with one anterior pituitary gland under the kidney capsule or were sham-operated, at 30 days of age. Both pituitary-grafted and sham-operated rats were injected subcutaneously with vehicle or CsA (5 mg/kg/day) for 9 days beginning at the 60th day of age. In pituitary-grafted male rats, mean prolactin levels, absolute prolactin pulse amplitude and mean half-life of the hormone increased, while the pulse frequency and relative amplitude of the peaks decreased, as compared with sham-operated rats. CsA administration to sham-operated rats decreased the relative pulse amplitude of prolactin but increased the mean half-life of the hormone, the pulse duration and the mean hormone levels, as compared with rats of the same group treated with vehicle. However, CsA treatment to pituitary-grafted rats decreased mean prolactin levels and the absolute amplitude of its peaks and increased the relative amplitude of its pulses, whereas all the other parameters showed no change. When considering circulating values of prolactin in plasma from the trunk blood, CsA administration was followed by changes similar to those described when mean values of the serial samples were considered. These data suggest the existence of an interrelationship between elevated circulating prolactin levels and CsA, which probably takes place at the hypothalamic level, to regulate the pulsatile pattern of prolactin secretion in male rats, although a direct effect of the drug on the ectopic gland in pituitary-grafted male rats cannot be excluded.

Urinary trehalase activity is a useful marker of renal proximal tubular damage in newborn infants.

To clarify the reliability of urinary trehalase activity as a marker of cellular proliferation and/or damage of renal proximal tubules, the activity was examined in healthy newborn infants or infants treated with tobramycin, a drug known as causing tubular cell damage. Eighty-one newborn infants (56 mature infants and 25 premature infants) were enrolled in the study. Urinary trehalase was examined using a spot urine sample during the first 7 days of age and on the 10th day of age. A good positive correlation was observed between urinary trehalase activity/creatinine ratio (T/Cr) on the 10th day of age and conceptional age or body weight (n = 46, r = 0.58, p < 0.001). Urinary trehalase of 29 healthy mature infants was higher during the first few days of age, after which it decreased to an almost steady level. Urinary trehalase of 6 premature infants during the first few days of age was significantly lower than that of mature infants, after which it increased and became equal to that of the mature infants on the 7th day of age. Treatment with ampicillin (100 mg/kg) and tobramycin (5 mg/kg) of 6 mature infants with pneumonia for 6 days resulted in a significant elevation of the urinary T/Cr. The extent of this elevation was greater than that of the urinary N-acetyl-beta-D-glucosaminidase (NAG) activity/creatinine ratio (NAG/Cr). A significant correlation was observed between the urinary T/Cr and the urinary NAG/Cr (r = 0.67, p < 0.01) or gamma-glutamyl transpeptidase/creatinine ratio (r = 0.48, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical Study on Localization of Tenascin in the Cervical Region of Rat Spinal Cord During Development.

Tenascin (TN) is transiently expressed in some organs and tumors at various developmental stages. It has also been reported that tenascin was a major extracellular matrix component in a developing nervous system, and played an important role in neurite outgrowth and pattern formation of the cerebral and cerebellar cortices.In the present study, we examined, under light and electron microscopes, the localization of TN in the cervical region of rat spinal cord during the late prenatal and early postnatal stages by means of conventional and immunohistochemical methods. In the specimens stained with hematoxylin-eosin and thionin, the following results were obtained: the structure of spinal cord was developing at the postnatal stages and attained a definite form around the 5th day after birth. Nissl bodies in neurons were maturing during this period and the processes of neurons elongated toward white matter. Concomitantly, neuroglia were also increased in the white matter.In accordance with it, luxol fast blue (LFB) and sudan black B stainable substances appeared in the white matter, especially ventral and lateral funiculi around the 5th day of age.The results of immuno-light microscopical observation showed that TN-labeling was observed only in the gray matter of the spinal cord at the 18th day of gestation, but both in gray and white matters from the 20th day of gestation to the 5th day after birth. The most intense TN-labeling was detected in the boundary of gray and white matter in the ventral and lateral funicili at the 2nd to 5th day after birth. As the development proceeded, up to the 2nd week after birth, TN-labeling was mainly distributed in the white matter except for the ventral region of dorsal funiculus and almost completely disappeared from the gray matter. Here, it was also noticed that the ventral region of dorsal funiculus became stainable with LFB or sudan black B between the 2nd and 3rd week after birth and at the same stage of development, TN-labeling became detectable in this region.At the electron microscopical level, TN-labeling was detected in the intercellular spaces of the gray matter during the 1st to 10th day of age. That is, TN was expressed associating with the cytoplasmic membrane and processes of neurons and neuroglia. TN-labeling was also observed along the cytoplasmic membrane of myelinating oligodendrocytes and in the cytoplasm of neuroglia, especially oligodendrocytes in the white matter at the 10th day to the 2nd week after birth. Clear reaction products for TN could not be detected in the synaptic region.From this evidence, it is concluded that TN mediates the interaction of neuron and neuroglia during late prenatal and early postanatal stages and plays an important role in elongation of neuronal processes and migration of neuroglia as well as the myelination in the developing rat spinal cord.

Early myelination in zitter rat: morphological, immunocytochemical and morphometric studies

Early myelination in zitter rat was investigated by light and electron microscopic observations, by immunostaining for myelin basic protein (MBP), proteolipid protein (PLP) and myelin-associated glycoprotein (MAG), and my morphometric analysis from the 1st to the 28th day of age. Although the commencement of myelination in zitter rats was not delayed in comparison with control rats, the density or the number of myelinated fibers in zitter rats was significantly below that in controls, both in the ventral column of the cervical spinal cord and in the optic nerve. In contrast, the density or the number of aberrant myelin sheaths was increased in zitter rats, and this difference became more marked with increasing age. The persistent presence of abnormal membranous structures associated with the oligodendroglial nuclear membrane and the increased number of aberrant myelin sheaths were characteristic to the zitter mutation, although these alterations were also observed transiently in cottrol rats. Quantitatibe analysis supported the proposition that hypomyelination in zitter rats is primarily pathological and becomes more prominent with advancing age. However, the fundamental structure of the myelin lamellae appeared to be normal and the immunoreactivities for MBP, PLP and MAG were slightly delayed and weakend in comparison with age-matched control. Thus, in zitter rat there is the fucntional abnormality of the oligodendrocytes to integrate the processes of membrane biosynthesis and to expel excessive production of membranous structures associated with the membranous organellae such as nuclaar membrane, and it is postulated that this functional abnormality is characteristic to only the zitter mutation.

Hypothalamic monoamines in the postnatal ontogenesis of rats

1. Sexual differences in the content of MA in the PMN, PLN, and AN in the early ontogenesis of rats have been found for the first time on the 7th day after birth, using histochemical methods. The sexual differences acquire a distinct character by the 60th day of life. 2. The intensity of the fluorescence of MA in the investigated hypothalamic nuclei of neonatally castrated male rats is maintained constantly reduced all the way to the 60th day of age. 3. The increase in the intensity of the fluorescence of MA in the PMN, PLN, and AN is a direct reaction to the neonatal virilization of female rats.

Effect of vitamin D supplementation during pregnancy on the neonatal skeletal growth in the rat.

In rats on normal intakes of calcium, phosphorus and vitamin D3, 3,000 and 7,500 IU of vitamin D3 were injected on the 10th day of pregnancy and the pups were investigated for the skeletal growth on 28th day of age. Compared to controls, the pups in the supplemented groups showed significantly greater dry weight and ash weight of the tibiae. However, the ash weight/dry bone weight ratios in the supplemented groups were not different from controls. Histological examination of the upper ends of decalcified tibiae and plasma calcium estimations revealed no abnormality in any group. The results suggest that vitamin D3 supplementation in pregnancy enhances the skeletal growth of the pups which involves both the organic and inorganic components.

Influence of rat prenatal and postnatal exposure to a non-steroidal anti-inflammatory agent (flunoxaprofen) on cardiovascular function in the progeny

The present experiments evaluated in rats the effects of prenatal and postnatal exposure to a non-steroidal antiinflammatory agent, flunoxaprofen (5–10 and 20 mg/kg/day by the oral route), on cardiovascular function in the pups. In both conscious and anaesthetized rats pre- and postnatal flunoxaprofen exposure at the 30th and 60th day of age, significantly (P < .05) induced a decrease of pressor response to carotid-sinus baroreceptor stimulation and to L-noradrenaline (0.1–1 and 5 μg/kg iv), and an increase of the hypotensive responses to L-isoprenaline (0.01–0.1 and 1 μg/kg iv) and acetylcholine (0.01–0.1 and 1 μg/kg iv). These effects were not observed in rats on the 90th day of age. Moreover, pre- and postnatal flunoxaprofen exposure did not modify systolic arterial blood pressure of plasma levels of catecholamines and acetylcholinesterases. Our results also show that in normotensive rats flunoxaprofen exposure during pregnancy did not affect the body weight, systolic or diastolic blood pressure or heart rate of pregnant rats. It did not affect the length of gestation, number of pups per litter or pup body weight. No macroscopic teratogenic effects were observed.

Factors contributing to cerebral hypomyelination in the growth hormone-deficientlittle mouse

We attempted to delineate the events leading to hypomyelination in the brain of the little mouse, a promising murine model of isolated growth hormone deficiency. At 20 days of age, the mutant mouse brain weighed less than its normal counterpart, and this difference in brain weight persisted. Increase in CNPase activity was found to be suppressed in the cerebrum throughout the developmental stage, but not in the other parts of the brain. Differences in cerebral DNA content between the little and normal mice first became apparent on the 10th day of age. Thereafter, the rate of increase in the little brain consistently lagged behind the normal. [3H]Thymidine incorporation into the DNA fraction in vivo on the 7th day of age, when glial cell proliferation in the normal cerebrum is most active, was approximately half that of the controls in all parts of the little brain. These findings indicate that the hypomyelination of the mutant cerebrum might result from reduced oligodendroglial proliferation due to growth hormone deficiency.

Humoral Immunity of Neonatal Swine after FMD Vaccination

The possible effect on pig protection after vaccination pregnant sows and their piglets against FMD at various age was examined using the SN test. Three experiments were conducted with three sow in each (8-9 piglets each). In the first experiment sows were not vaccinated but their piglets were vaccinated on 10th, 20th and 60th day of age. In second experiment sows were vaccinated at the end of rest period and in the middle of pregnancy. Pigs from one sow were vaccinated on 10th, from second sow on 20th and from the third sow on the 60th day of age. Pigs which were vaccinated on the 10th and 20th day of age were revaccinated on the 60th day of age. In third experiment sows were vaccinated at the end of the rest period and in the middle of pregnancy. Their piglets were not vaccinated. The conclusion may be drawn that if the epidemiological situation requires, systemic vaccination of all pigs then, on large farms (where pig production is planned) all sows should be vaccinated at the end of the rest period (first vaccination) and on the 55th-60th day of pregnancy. The next vaccination should be done in the middle of the next pregnancy. A vaccination program of piglets, according to our results, should depend on the vaccination program used for their dams.

Effect of vitamin D administration during pregnancy on neonatal growth in the rat.

Groups of rats on commercial diet were injected 3,000 IU (group 2) and 7,500 IU (group 3) of vitamin D3 on the 10th day of pregnancy. Compared to control pups (group 1), the pups in group 2 and 3 weighed significantly more on the 10th, 20th and 28th day of age. At 28th day of age, study of the gastrocnemius muscle revealed significantly greater organ weight, protein, DNA and RNA contents, protein/DNA and RNA/DNA ratios in group 2 and 3 pups than in controls. In the liver, whereas all these indices were significantly increased in group 3 pups, only protein/DNA and RNA/DNA ratios were significantly increased in group 2 pups. Brain weight, its RNA content and RNA/DNA ratio were significantly greater in group 2 and 3 pups than in controls. The results suggest that vitamin D administration in pregnancy promotes soft tissue growth in the pups by enhancing cellular proliferation and hypertrophy.

Interference of prenatal and postnatal exposure to Ca2+‐antagonist agents on rat functional development of vascular system

Exposure to drugs during pregnancy can alter functional development of the vascular system. The present investigation was carried out in order to evaluate the effects of prenatal and postnatal exposure to Ca2+-antagonist (diltiazem, verapamil, and nimodipine) drugs on the development of rat vasomotor reactivity. Studies were carried out on pregnant female albino rats exposed from the first day of pregnancy until weaning to diltiazem and verapamil (6 and 24 mg/kg in their drinking water ad libitum) and nimodipine (3 and 12 mg/kg in their food ad libitum). After weaning, pups were exposed until the 60th day of age to the same treatment as their mothers were. Afterwards, pups from the 60th to 90th day of age were fed with a normal diet. In 30-, 60-, and 90-day-old conscious and anaesthetized pups, we evaluated the following: 1) systolic arterial blood pressure; 2) vasomotor responses elicited by various agents: L-noradrenaline (0.1, 1, and 5 micrograms/kg IV), L-isoprenaline (0.01, 0.1, and 1 micrograms/kg IV), and acetylcholine (0.01, 0.1, and 1 micrograms/kg IV) and by sinus-carotid baroreceptor stimulation; and 3) catecholamine, acetylcholinesterase, and adenosinase plasma levels. Prenatal and postnatal exposure to Ca2+-antagonist drugs significantly (P less than .05) decreased the pressor response to sinus-carotid baroreceptor stimulation and to L-noradrenaline and increased the hypotensive responses to L-isoprenaline and acetylcholine. Moreover, this type of treatment, although it induced a significant (P less than .05) decrease of catecholamine plasma levels, did not modify the acetylcholinesterase and adenosinase plasma levels in 30- and 60-day-old rats. On the 90th day of age, the evaluated parameters were not different from those of control rats. Our results showed that exposure to Ca2+ antagonists during pregnancy and the postnatal period may alter the functional development of rat vasomotor reactivity.

High Dose Nicotinamide Fails to Prevent Diabetes in BB Rats

Nicotinamide which is an inhibitor of poly (ADPR) synthetase and precursor of NAD has been observed to prevent diabetes in some experimental models possibly by protecting beta cells. To determine whether nicotinamide could cure or prevent type 1 diabetes, we administered large doses (0.5 g/Kg/d) to BB rats. When used in the 45 days following diagnosis nicotinamide failed to bring remission. As a preventive treatment, nicotinamide administered between the 40th and 90th day of age, alone or in association with desferrioxamine did not significantly lower the incidence of diabetes (23% and 30.8% respectively vs. 56.6%). When used earlier, immediately after weaning, nicotinamide did not affect the incidence of diabetes in this model (62.5%). The degree of protection was not comparable with that obtained with cyclosporin A (15% of diabetic animals). Histology study of the pancreas from the animals killed either immediately or 1 year after treatment revealed no endocrine tumor. These findings suggest that in BB rats nicotinamide has little or no effect on the course of autoimmune diabetes mellitus thus dampening the high hopes for this drug in the treatment of human diabetes.