Dynamics of microcirculatory system recovery in the post-stress period

Twenty-four-hour immobilization or electric stimulation for 6 h were used in rats as stressors. The first stressor caused more profound and protracted disturbances in the microcirculatory system. The recovery of the microcirculation occurred in 50% of animals by day 6 and in 100% by day 14 after immobilization. The terminal blood flow recovery after 6-hour electric stimulation was seen in a day. Vascular permeability after 24-hour immobilization returned to normal in 24 h, and after 6 h of electric stimulation in 6 h. This process correlated with the morphofunctional status of mast cells and was probably phasic in nature.     

The effects of melatonin, N-acetylserotonin, and 6-hydroxymelatonin on the ultrastructure of the pinealocytes of the Syrian hamster (Mesocricetus auratus)

OBJECTIVES: The aim of this study was to examine the effects of melatonin as well as of its precursor (N-acetylserotonin) and metabolite (6-hydroxymelatonin) on the ultrastructure of the pinealocytes of the Syrian hamster. MATERIAL AND METHODS: The pineal glands of 2-month-old male Syrian hamsters were examined. The animals were divided into the following groups of four animals each: group 1 - melatonin treatment; group 2 - N-acetylserotonin treatment; group 3 - 6-hydroxymelatonin treatment (all substances given subcutaneously at doses of 25 microg per animal between 16.00 and 17.00 h daily for seven weeks). Group 4 was given solvent treatment only and served as controls. The animals were killed by decapitation between 09:00 and 10.00 h. Routine electron microscopical techniques were used to obtain quantitative data on pinealocyte ultrastructure. RESULTS: Melatonin administration did not influence the size of the hamster pinealocytes, whereas administration of N-acetylserotonin and 6-hydroxymelatonin caused a significant reduction in cell size in comparison to the melatonin-treated and control groups. There were changes in the relative volumes of the mitochondria, Golgi apparatus and lysosomes in the pinealocytes of the studied groups, while the volumes of granular endoplasmic reticulum and lipid droplets were unchanged. The dense-core vesicles were more numerous in the pinealocytes of the melatonin and 6-hydroxymelatonin-treated groups in comparison to those of animals treated with N-acetylserotonin or the controls. CONCLUSIONS: The changes observed in the ultrastructure of hamster pinealocytes indicate that administration of melatonin as well as of its precursor or metabolite influences the morphology of these cells and also, perhaps, their secretory activity.     

Detailed profile of prolactin secretion in the immature female rat: evidence for the existence of an ultradian rhythm

The detailed profile of prolactin (PRL) secretion in 22-24 and 29-31 days old female rats was investigated by serial blood sampling through an intracardiac cannula at 15-min intervals for each of the 9 or 10-h periods beginning at 09.00 or 10.00 and 22.00 h. By analysis of the power spectrum and the least squares method the time series of PRL concentrations which were measured by RIA were found to have approximately a 3-h period ultradian rhythm in either sampling period of both the 22-24 and 29-31 days old rats. The peak times calculated based on the acrophase estimated through the calculation of periodicity were concentrated around 12.00, 15.00 and 18.00 h for the sampling period 10.00-19.00, and 24.00, 03.00 and 06.00 h for the sampling period 22.00-07.00 h. However, in more than half of the animals at 22-24 days of age, one secretory episode around 12.00 h, and two secretory episodes around 24.00 and 03.00 h had markedly small amplitudes, making the remaining secretory episodes distinct diurnal and nocturnal surges, respectively. In the animals at 29-31 days of age, the amplitudes of the PRL episodes occurring around 12.00 h were markedly small, making the remaining two episodes as diurnal surges, whereas the amplitudes of PRL secretory episodes during the period 22.00-07.00 h were analogous to each other. These findings indicate that the semicircadian rhythm of PRL secretion is established on the basis of PRL secretion with the 3.0-h period ultradian rhythm.     

Anti-stress effect of dalargin in immobilization stress in rats

The function of the isolated perfused rat hearts was studied in four groups of experiments. Group 1--included the hearts of intact animals ("absolute control"), group 2--the hearts of rats subjected to 24 hour immobilization in supine position against the background of triple intramuscular injections of placebo (control), group 3 included the hearts of rats, which during 24 hour immobilization stress were thrice injected a synthetic analogue of endogenous opioids Dalargin in a dose of 3 g/kg, and group 4 included the hearts of animals, which during immobilization were administered Dalargin in a dose of 10 g/kg of body mass. Ulcer index as indicator of stress injury of gastric mucosa was also determined. In the control group of experiments (group 2) 24 hour immobilization stress resulted in complete depression of cardiac performance as compared with group 1, and ulcer index approximated 1. In group 3 the indices of cardiac performance even exceeded those in group 1 (intact animals). As compared with group 2, ulcer index in group 3 decreased by 9 times. In application of Dalargin in a dose of 10 g/kg complete preservation of heart function indices and complete prevention of stress injury of gastric mucosa were also observed. Thus, Dalargin possesses cardioprotective and anti-ulcerogenic effect in immobilization stress in rats. Most probably, this phenomenon can be attributed to its ability to inhibit the activity of sympathoadrenal system, which gets enhanced during stress.     

Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows

The aim of the present study was to confirm earlier findings, obtained with a small number of animals, that gonadotropin-releasing hormone (GnRH) can shorten corpus luteum functional life when it is administered 24 h after cloprostenol (PG) treatments given 7-9 days after estrus. In addition, the effects of two treatments, PG alone or PG + GnRH given before mid-diestrus, on signs of estrus were studied. Sixty cows in farm conditions were used in the experiment. Eight days after natural estrus, they were given an intramuscularly (i.m.) treatment of cloprostenol (0.5 mg). The animals were then divided into two groups. One group (n = 25) received an i.m. treatment of gonadorelin (0.1 mg) 24 h after the PG treatment (PG + GnRH group), while another group (n = 35) served as controls without any further treatment (PG group). Estrous signs were recorded. Progesterone concentrations were measured from samples of whole milk. No short cycles were observed in the PG group, whereas 33% of the cows in the PG + GnRH group exhibited premature luteal regression (P < 0.05). Cloprostenol treatment on Day 8 had no effect on the intensity of the estrous signs. Instead, GnRH treatment 24 h after PG treatment weakened the estrous signs significantly (P < 0.01). It is concluded that GnRH administration 24 h after a PG treatment given 8 days after estrus can cause short estrous cycles in some cows on an individual basis.     

Hibernation and disuse osteoporosis.

In an attempt to determine 1) if disuse osteoporosis occurs naturally during hibernation and 2) if it can be induced by limb immobilization in active and hibernating ground squirrels, forty-six thirteenlined ground squirrels were divided into five experimental groups. Group one was the active control group (AC) (ten animals). Group two was the active immobilized (AI) group and was subjected to a unilateral sciatic nerve section (ten animals). Group three was the hibernating control group and was allowed to hibernate undisturbed for 30 days (HC) (ten animals). The fourth group was subjected to a unilateral sciatic nerve section and allowed to hibernate for 30 days; this was the hibernating immobilized group (HI). The fifth group of six animals received the identical treatment as the (HI) group but was allowed to hibernate for 150 days.The following measurements were taken: Gastrocnemius-soleus mass, body mass, tibia-fibula weight, total tibia-fibula calcium and phosphorus. In addition, the femur was observed histologically for the enlarged lacunae typical of osteoporotic bone. Statistical analysis was accomplished with the Wilcoxan sign rank test.The data were interpreted to mean: 1) Disuse osteoporosis can be induced in active ground squirrels, 2) disue osteoporosis is not evident nor can it be induced by limb immobilization during 30 days of hibernation and 3) limb immobilization did not result in significant disuse osteoporosis following 150 days of hibernation. Only a few enlarged lacunae were evident after 150 days of hibernation.     

Plasma cortisol in cattle: Circadian rhythm and exposure to a simulated high altitude of 5,000 m

After a 2 week control period at 400 m, cattle were exposed to 5,000 m simulated altitude for 2 weeks, which was followed by a 2-week post-altitude control period. Plasma cortisol values from blood samples taken every 30 min for a total of 24 h indicated that cortisol was secreted episodically and that a circadian rhythm existed. When cortisol values were grouped into 4, 6-h periods, plasma cortisol was most abundant from 06:00 to 12:00 h with an average of 0.96µ g/100 ml and least abundant from 00:30 to 06:00 h with an average of 0.55µ g/100 ml. Plasma cortisol increased from 0.42 to 3.08µ g/100 ml during the 4 h ascent to 5,000 m and decreased to near normal levels the following day. A rhythmic plasma cortisol pattern was maintained after one day at simulated high altitude.     

Changes in the adrenal medulla of the rat during immobilization stress

A combined histofluorescent, light optic and electron microscopic investigation has been performed to study the medulla of the adrenals in intact rats (August strain) and immediately after immobilization for 30 h and 24 h after. In the intact animals heteromorphism of the medulla is demonstrated; this depends on the fact that adrenocytes are at different stages of the secretory cycle. Immobilization for 30 h results in synchronization of secretion; this is determined by adaptation of the adrenal system to immobilization. One day after immobilization restoration of heteromorphism in chromaffinocytes is demonstrated. The changes described are compensatory-adaptive reactions of the adrenal medulla to the stress in the animals survived.     

Modifications in the oscillation of substrates in obese subjects subjected to variations in the pattern of meal-timing

In a group of obese subjects kept on hypocaloric diet during a period of 18 days, an eventual different behaviour of carbohydrates and lipids oxidation following to meal-timing manipulation (a single meal given only at 10.00 hr or at 18.00 hr of each day) has been studied by indirect calorimetry. No significant difference of body weight reduction was noticed in both dietetic regimens. In the group of obese subjects who consumed the single meal at 10.00, a slight increase of carbohydrates oxidation was noticed, without a statistical significance, whereas in the group with single meal at 18.00 h a significant increase of lipids oxidation resulted.     

Serotonergic immunoreactivity in the pedal ganglia of the pulmonate snail Megalobulimus abbreviatus after thermal stimulus: A semi-quantitative analysis

Using an immunohistochemical procedure and optical densitometry, the distribution of neurons containing serotonin (5-HT) was investigated in the pedal ganglia of Megalobulimus abbreviatus after thermal "non-functional stimulus" (22 degrees C) and stressful thermal conditions (50 degrees C).The animals were sacrificed at different times (3 h, 6 h and 24 h) following these stimuli. In control animals, the results showed the location of these serotonergic immunoreactive elements (5HT-ir) in this ganglion to be similar to those shown in other studies, where the anterior region of ventral sections showed the largest number of 5HT-ir neurons. In the anterior neurons, significant differences (p < 0.01) were observed between the groups of animals stimulated at 50 degrees C and 22 degrees C and sacrificed after 6 h. In the medial neurons, significant differences (p < 0.05) were observed between the control group and the groups of animals stimulated at 50 degrees C and sacrificed after 6 and 24 h. Neuropilar area 1 showed differences (p < 0.01) in 5HT-ir between the control group and the groups of animals stimulated at 50 degrees C and sacrificed after 3 and 24 h. Neuropilar area 2 showed a significant difference (p < 0.05) between the groups of animals stimulated at 22 degrees C and sacrificed after 3 and 24 h. These results suggest the involvement of 5-HT in the nociceptive circuit of M. abbreviatus, mainly that of the medial neurons and neuropilar area 1, which showed increases in 5HT-ir after thermal aversive stimuli. These results could be helpful in drawing cellular homologies with other gastropods.     

Stress-induced change in cerebral ATP+ubiquitin-dependent proteinase activity vary with age.

Stress-induced changes in cerebral ATP+ubiquitin-dependent proteinase activity were studied and the effect of age on it was checked. For that purpose 23,000 g supernatant prepared from whole brain of 3- and 7-month-old rats after 6h long immobilization stress were used. With azocasein as substrate, at pH 8.0 values of ATP+ubiquitin-dependent proteinase activity increased for 20% and 10% in 3- and 7-month-old animals respectively. Following 24 h long immobilization, values of cerebral ATP + ubiquitin-dependent activity in 3-month-old animals dropped by 16%. Data obtained indicate that immobilization stress affects ATP+ubiquitin-dependent proteinase activity and point to the contribution of age in the modulation of enzyme response to stress.     

Escherichia coli K12 does not colonize the gastrointestinal tract of Fischer-344 rats

Summary The colonizing potential ofEscherichia coli K12 containing a vector coding for somidobove (bovine somatotropin) was determined. Treated male and female Fischer-344 rats were given a single oral gavage inoculum of sucrose with/without tetracycline (15 g/ml). Untreated control animals received similar drinking water regimes. All animals survived until termination. There were no clinical signs of toxicity observed and no treatment-related effect upon body weight, food consumption, or efficiency of food utilization. Fresh fecal samples were collected from each rat every 24 h following inoculation and the population of the marked strain was quantitated until no bacterial colonies were observed for two consecutive days. While all inoculated rats were positive at 24 h, by 72 and 96 h all had become negative for the test (marked) strain, as were the corresponding control group throughout the test. The frozen stock of the marked strain used as the positive control demonstrated that the agar plates were selective for the test strain. Fourteen days following inoculation, all groups of rats were killed and the gastrointestinal tracts removed and treated to recover the marked strain. There was no evidence of the marked strain in the gastrointestinal tract of any rat from any group. Thus, theE. coli K12 host/vector system used in this experiment does not colonize the gastrointestinal tract of Fischer-344 rats.     

Ultrasound measurement of the volume of musculus quadriceps after knee joint injury

The monitoring of the recovery of femoral muscles, after the knee-joint injury, is possible by the method of ultrasound measurement of the muscular volume. In a clearly defined longitudinal study, our object was to standardize the method of ultrasound measurement of muscular volume and to evaluate its adequacy in practical application in quadriceps muscle rehabilitation. The ultrasound measurements of m. rectus femoris and m. vastus intermedius were conducted in three intervals: in the first 24 hours after the injury; after 1 week, when immobilization was removed; and after 6 weeks, when rehabilitation was finished. The study comprised 30 patients with knee-joint injury, and 30 asymptomatic subjects, who formed the control group. The results showed significant decrease of muscular volume (mm3) after joint immobilization on injured leg and a significant increase of volume after rehabilitation. The same differences were observed on healthy legs, but without significance. Within the same intervals, there were no changes in the muscular mass in the control group. M. rectus femoris was completely recovered in greater number of patients (54.1%), comparing to m. vastus intermedius (25.4%). We conclude that the ultrasound is an appropriate method for monitoring the process of muscular atrophy during immobilization, as well as the course of muscular restitution during the physical therapy.     

An exploratory study of behavioural differences between young pigs susceptible and non-susceptible to stress syndrome

The behaviour of eight stress-susceptible and eight normal juvenile pigs was compared by visual observation to test the possibility for pre-determining, to some degree, the stress susceptibility on behavioural grounds. Data were analysed according to observation time and the physiological status of each animal. During the feeding period (08.00–10.00 h), the general level of activity of all the pigs was higher, as expected; 13 of 20 behaviours were more frequent.From 08.00 to 10.00 h, stress-susceptible animals drank more often and investigated the environment and their peers by naso-ear and naso-anal contacts more frequently than normal animals, while they were seen lying less often. However, frequency of all behaviours was similar from 10.00 to 12.00 h. It is suggested that behavioural recognition of stress-susceptible swine could be a simple, reliable and non-stressful method, although it still remains complementary to physiological detection.     

The individual postradiation reactions of cyclic nucleotides in the lymphoid cells of the rat spleen]

X irradiation (4.3 Gy) of rats was shown to cause different reactions of the cyclic nucleotide system in differently reactive animals which was determined by the neutrophilic response in the peripheral blood after three-hour immobilization. Changes in the cyclic nucleotide content and in the activity of adenylate cyclase were found to occur in both hyper- and hyporeactive animals 24 h after irradiation. In hyperreactive animals, however, these changes were more pronounced: a relative ability of adenylate cyclase to be activated in hyperreactive animals made 30%, and 52% in hyporeactive animals. The cAMP content in lymphocytes of the spleen of hyporeactive rats was twice as high as that in hyperreactive animals.     

NADPH-diaphorase expression in the rat jejunum after intestinal ischemia/reperfusion

The purpose of this study was to analyze the nicotinamide adenine dinucleotide phosphate - diaphorase (NADPH-d) activity in the rat jejunum after a mesenteric ischemia/reperfusion injury. Nitric oxide, synthetised from L-arginine by the enzyme nitric oxide synthase, is a nonadrenergic noncholinergic relaxant neurotransmitter of the intestinal smooth muscle. It plays an important role in the process of plasticity after the ischemia/reperfusion injury. Experimental animals were divided in two groups: the control group and the ischemic/reperfusion group, with different period of the reperfusion. The NADPH-d histochemical method has been used as a marker for the nitric oxide synthase. NADPH-d activity has been rapidly decreased in the neurons of both enteric nervous systems in plexuses of the jejunum after 1 h mesenteric ischemia and 1 h reperfusion. Differences were predominantly detected in the myenteric plexus; they were seen in change of the neuronal shape, in the arrangement of neurons and in intensity of their staining. The NADPH-d positivity was absent in the intestinal crypts. After 1 h ischemia and 24 h reperfusion, the NADPH-d activity was gradually increased, but it was lower in comparison with the control group. On the 30th day following the ischemia/reperfusion there were no changes in NADPH-d positivity compared with the control animals. These results indicated that the jejunal ischemia/reperfusion has affected the neurons of the enteric nervous system of adult rats and resulted in the early decrease of NADPH-d positivity 1 h of the reperfusion insult. The gradual increasing of NADPH-d activity in 24 h following the reperfusion could be considered as a result of the plasticity process. On the 30(th) day after the ischemia/reperfusion all histochemical changes were returned to the control levels.     

Effects of intracerebroventricular infusion of genistein on the secretory activity of the GnRH/LH axis in ovariectomized ewes

Phytoestrogens, plant derived estrogen like-compounds exert numerous effects on the reproductive functions of animals. The present study was designed to demonstrate if exogenous genistein infused during the breeding season into the third ventricle of the brain of ovariectomized ewes could affect the secretory activity of the GnRH/LH axis. Two-year-old ovariectomized ewes (n=8) were infused with vehicle (control, n=3) or genistein (10 microg/100 microl/h, n=5) into the third ventricle. The infusions were done from 10.00 to 14.00 h and blood samples collection was performed this day up to 20.00 h and next day from 8.00 to 10.00 h. The animals were slaughtered, thereafter. Immunoreactive (IR) GnRH neurons in the hypothalamus and LH cells in the adenohypophysis were localized by immunohistochemistry. Messenger RNA analyses were performed by nonisotope in situ hybridization using sense and anti-sense riboprobes produced from beta subunits of LH cDNA clones. Plasma LH concentrations were measured by radioimmunoassay. Immunohistochemical analysis revealed that genistein infusion affected the morphology of GnRH neurons evoking a visualization of long axons in the GnRH perikarya and visibly diminished IR GnRH stores in the median eminence. The number of IR LH cells and IR material stored in the adenohypophyses increased in genistein-infused animals, which was confirmed by statistical analysis (P<0.001). The in situ hybridization analyses showed in these ewes the increase of mRNA LHbeta hybridization signal. The changes in LH release in response to genistein infusion had a biphasic character: it decreased within 6 h after infusion and increased 24 h later. Mean concentration of LH and amplitude of pulses measured from the beginning of infusion up to end of the experiment were significantly higher (P<0.05) in genistein-infused ewes compared to vehicle-treatment. In conclusion, our data show that genistein, a phytoestrogen, may effectively modulate GnRH and LH secretion in OVX ewes by acting directly on the CNS. The biphasic character of the LH response is similar to that of estradiol during the breeding season in the ewes.     

Effect of a meal feeding schedule on hepatic glycogen synthesis and gluconeogenesis in rats

We investigated the effect of a meal feeding schedule (MFS) on food intake, hepatic glycogen synthesis, hepatic capacity to produce glucose and glycemia in rats. The MFS comprised free access to food for a 2-hour period daily at a fixed mealtime (8.00-10.00 a.m.) for 13 days. The control group was composed of rats with free access to food from day 1 to 12, which were then starved for 22 h, refed with a single meal at 8.00-10.00 a.m. and starved again for another 22 h. All experiments were performed at the meal time (i.e. 8.00 a.m.). The MFS group exhibited increased food intake and higher glycogen synthase activity. Since gluconeogenesis from L-glutamine or L-alanine was not affected by MFS, we conclude that the increased food intake and higher glycogen synthase activity contributed to the better glucose maintenance showed by MFS rats at the fixed meal time.     

Liver glycogen content and activity of several hepatic enzymes in repeatedly immobilized rats

Liver glycogen content and activity of several hepatic enzymes in repeatedly immobilized rats. Acta Physiol. Pol., 1978, 29 (1): 55-60. In adult, male rats subjected to immobilization for 2.5 h daily, on the 7th day, 24 h after a sixth trial, an increased activity of the liver aspartate aminotransferase was found as shown by comparing their data with those of normally fed controls and also of the animals pair fed to the immobilized. Moreover, a net increasing effect of daily immobilization on the liver glycogen content was described.