The effects of perindopril on vascular smooth muscle polyploidy in stroke-prone spontaneously hypertensive rats

OBJECTIVE: To quantify vascular smooth muscle polyploidy and growth kinetics in aortic cells from stroke-prone spontaneously hypertensive rats (SHRSP) and from normotensive Wistar-Kyoto (WKY) rats, and to examine the effects of treatment with the angiotensin converting enzyme (ACE) inhibitor perindopril on these parameters. DESIGN: The following experimental groups were used: young (age < 20 weeks) and old (age > 20 weeks) untreated WKY rats and untreated SHRSP; SHRSP treated with perindopril, and age- and sex-matched control SHRSP; and SHRSP treated with hydralazine and hydrochlorothiazide and age- and sex-matched control SHRSP. The effects of treatment of the SHRSP with perindopril for 30 days on vascular smooth muscle polyploidy and growth kinetics were measured and compared with the effects of equivalent antihypertensive doses of hydralazine and hydrochlorothiazide. METHODS: Vascular smooth muscle polyploidy was measured using flow-cytometry DNA analysis of freshly harvested cells. Growth curves were performed on cultured aortic cells. Plasma renin activity was measured by an antibody-trapping method, plasma angiotensin II (Ang II) by radioimmunoassay and plasma ACE activity by a colorimetric method. Cardiac hypertrophy was evaluated by measuring the heart weight:body weight and left ventricle + septum weight:body weight ratios. RESULTS: The SHRSP had markedly and significantly elevated G2 + M phase of the cell cycle. Treatment with perindopril resulted in a significant reduction in polyploidy in the SHRSP, whereas treatment with hydralazine and hydrochlorothiazide had no effect on the percentage of cells in the G2 + M phase of the cell cycle. The regression of polyploidy after treatment with perindopril was associated with a significant reduction in the concentration of Ang II and ACE activity, and with a significant regression of cardiac hypertrophy. Increased mitogenesis of cultured vascular smooth muscle cells from the SHRSP was not altered by treatment with perindopril. CONCLUSIONS: ACE inhibition reduces vascular smooth muscle polyploidy in large conduit arteries. This type of vascular protection is mediated by the reduced Ang II and possibly by increased kinins level, rather than by the hypotensive effect alone.     

A comparison of the effects of feeding concentrate diets, based on either maize or barley, or dried grass on adipose tissue lipogenesis in sheep

1. Three groups of 4 sheep were penned individually and provided with diets composed of either dried grass, 80% ground maize/20% soyabean meal or 80% ground barley/20% soyabean meal. 2. The diets were fed ad libitum for 3 weeks before s.c. adipose tissue biopsy samples were taken from the rump region. 3. Although the rate of lipogenesis was significantly increased by concentrate feeding the order of utilization of the various substrates was always acetate greater than glucose greater than lactate. Throughout this work lactate was always of minor significance as a lipogenic substrate. 4. The diet-induced differences in lipogenesis were reflected in significant increases in the specific activities of the fatty acid synthetase system, glucose-6-phosphate dehydrogenase and phosphogluconate dehydrogenase in concentrate fed animals. 5. No differences were observed in in vitro lipogenesis from any of the substrates or enzyme specific activity between the 80% barley diet and the 80% maize diet. 6. These results are discussed in relation to the effect of concentrate and roughage feeding on the entry of alpha-linked glucose polymer into the small intestine of sheep.     

BCG infection suppresses allergic sensitization and development of increased airway reactivity in an animal model

We previously demonstrated that chronic dietary treatment with acarbose, an alpha-glucosidase inhibitor, improves glucose homeostasis in the streptozotocin (STZ)-induced diabetic rat. In this study we evaluated the effects of 4 weeks of acarbose treatment on glucose homeostasis in STZ-diabetic rats for both meal-fed (three times daily) and ad libitum feeding conditions. Sprague Dawley male rats (n = 58) were started on a daily meal-feeding paradigm consisting of three 2-h feeding periods: 0700 to 0900 hours, 1300 to 1500 hours, and 1900 to 2100 hours. Following 2 weeks of adaptation, half of the animals were switched to ad libitum feeding. The feeding paradigm itself (meal fed versus ad lib.) affected neither body weight nor daily food intake. Twenty animals from each feeding group then received STZ (60 mg/kg i.v.), whereas control animals received vehicle injections only. Two days later, the diet of 10 STZ-treated animals from each paradigm was supplemented with acarbose (40 mg of BAY G 5421/100-g diet), and the groups were treated for 4 weeks. Untreated diabetic rats had lower body weight than vehicle-injected control rats at all time points after STZ treatment. Acarbose treatment delayed this effect on body weight. STZ treatment induced hyperphagia regardless of feeding paradigm, which was significantly attenuated by acarbose only for the first week of treatment. Untreated diabetic rats had fasting blood glucose values 4 times those of vehicle-injected controls in both the meal-fed and ad libitum-fed conditions. Acarbose significantly lowered fasting blood glucose in the treated STZ groups. Blood glucose was also assessed 0, 90, and 180 min following the start of a meal. The postprandial rise in blood glucose was significantly reduced in acarbose-treated meal-fed diabetic rats, to values not significantly different from those of vehicle-injected control rats. During the fourth week of treatment glycated hemoglobin levels were significantly higher in untreated diabetic groups compared to vehicle-injected control groups. Acarbose treatment significantly reduced this rise, regardless of the feeding paradigm. Collectively, the results demonstrate that acarbose reduces diabetes-induced increases of blood glucose and glycated hemoglobin and that the glycemic effects of acarbose are most apparent during the absorptive period. Feeding paradigm (ad lib. versus meal fed) has little or no influence on acarbose's metabolic effects, indicating that large meals are not required to realize the beneficial effects of the drug. The meal-fed STZ-diabetic rat may be a good model with which to test meal-based diabetes treatments.     

Rearrangement of the ventricular capillary network in stroke-prone spontaneously hypertensive rats (SHRSP) following a late start of treatment with the angiotensin converting enzyme blocker temocapril

The effects of the angiotensin converting enzyme (ACE) blocker temocapril on the capillary network of the left ventricle were studied in stroke-prone spontaneously hypertensive rats (SHRSP). The ACE blocker was dissolved in the drinking water and supplied to 24 and 32 week old SHRSP ad libitum for 5 weeks. The capillaries of the wall of the left ventricle were studied using a double staining method to differentiate the arteriolar, intermediate and venular capillary portions. Capillary density increased and capillary domain areas decreased in all capillary portions compared with untreated control SHRSP in both age groups. The proportion of venular capillary portions was increased by temocapril treatment. The results indicate that the late start of ACE blockade caused the regression of the hypertrophied cardiomyocytes, which is characteristic of SHRSP, and the rearrangement of capillary portions. The plasma concentration of angiotensin II was significantly lower in temocapril-treated SHRSP compared to the control group. The implication is that intrinsic angiotensin II exerts an appreciable effect on the function, structure and capillary network in the left ventricular wall in SHRSP.     

Endoscopic repair of mandibular subcondylar fractures

To investigate effects of sustained activity on major phenotypic properties, the left extensor digitorum longus muscle of young (15 wk) and aging (101 wk) male Brown Norway rats was subjected to 50 days of chronic low-frequency stimulation (CLFS; 10 Hz, 10 h/day). The contralateral muscle served as control. Changes in metabolic enzymes were analyzed by using glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase as reference enzymes of glycolysis and by using citrate synthase and 3-hydroxyacyl-CoA dehydrogenase as mitochondrial enzymes representative of aerobic-oxidative metabolism. Myosin heavy chain (MHC) isoforms were analyzed by SDS-PAGE. No differences existed between the enzyme activity profiles of control muscles from young and aging rats. CLFS induced similar increases in mitochondrial enzymes, as well as similar decreases in glycolytic enzymes. Although the MHC composition of the control muscles in the aging rats displayed a shift toward slower isoforms, the ultimate changes induced by CLFS led to nearly identical MHC phenotypes in both young and aging rats. These results demonstrate an unaltered adaptability of skeletal muscle to increased neuromuscular activity in the aging rat.     

Food for inefficient thought

Sixty Sprague-Dawley rats were fed diets containing 0, 10, 100, 1,000, and 10,000 ppm of methoxychlor for 16 weeks under ad libitum- and restricted-feeding regimens. Methoxychlor at 10,000 ppm was lethal to some rats, reduced food consumption and growth, and increased liver weight relative to body weight. Methoxychlor at 1,000 ppm reduced food consumption and growth of rats fed ad libitum but did not reduce growth of restricted-fed rats. Reduced hepatic storage of vitamin A was detectable when methoxychlor was fed at levels of 100 ppm or higher.     

Luminal factors stimulate intestinal repair during the refeeding of malnourished infant rabbits

We examined the role of luminal versus systemic factors in promoting intestinal recovery during the refeeding of previously malnourished infant rabbits. Malnutrition was induced by litter expansion at 7 days of age. A 20-cm Thirty-Vella (T-V) loop was created in the intestine of each malnourished and dietary control animal at 21 days of age. Beginning on day 28, controls and one half of the malnourished group (malnourished-refed) were fed chow ad libitum, whereas the remainder of the malnourished group received half the amount of chow given to the malnourished-refed group. On day 35, proximal and distal segments from the intact intestine that remained in continuity as well as segments from the excluded T-V loops were examined. Malnutrition severely reduced mucosal mass and disaccharidase activities in the intact distal intestine. A brief period of refeeding led to a rapid recovery of these parameters. In contrast, the excluded T-V loop segments of the control, malnourished, and malnourished-refed groups all displayed decreased mucosal mass and impaired function to a degree similar to that observed in the intact distal segment from the malnourished group. These results indicate that luminal factors are essential for (i) the maintenance of normal intestinal structure and function in infant rabbits and (ii) the promotion of mucosal repair following nutritional rehabilitation of malnourished animals.     

Differential alterations in left and right ventricular G-proteins in congestive heart failure due to myocardial infarction

In order to examine the status of G-proteins in congestive heart failure due to myocardial infarction, the left coronary artery in rats was ligated and animals assessed after 4, 8 and 16 weeks. Sham-operated control and experimental animals were used for the preparation of membranes from the viable (uninfarcted) left and right ventricles. Adenylyl cyclase activities in the presence of pertussis toxin and cholera toxin were increased and decreased in left ventricles from all groups, respectively. On the other hand, adenylyl cyclase activities in 8 and 16-week experimental right ventricles were unaltered in the presence of pertussis toxin and increased in the presence of cholera toxin. Depression of adenylyl cyclase activities in left ventricles from all groups as well as in the right ventricle at 4 weeks were not evident when enzyme activity was determined in the pertussis toxin-treated membranes in the absence or presence of Gpp(NH)p. Cholera toxin-catalyzed ADP ribosylation was decreased in left ventricles from all infarcted groups and increased in the right ventricles at 8 and 16 weeks whereas the pertussis toxin-catalyzed ADP ribosylation was increased in all experimental tissues except in the right ventricles at 8 and 16 weeks. G(s alpha)-protein content was decreased in the left ventricle at 16 weeks and increased in the right ventricles at 8 and 16 weeks of myocardial infarction. On the other hand, G(i alpha)-protein content was increased in left ventricles from all infarcted groups and the 4-week right ventricle but was unaltered in 8 and 16-week right ventricles. An increase in mRNA abundance for G(i alpha)-protein was seen in both left and right ventricles following myocardial infarction. A significant increase in mRNA level for G(s alpha)-protein was observed in all left ventricles and 8-week right ventricle following the coronary occlusion. These results suggest that changes in Gs- and Gi-proteins in the failing heart due to myocardial infarction are chamber-specific and are dependent upon the stage of congestive heart failure.     

Effect of feeding regimen on gastric ulceration of the pars oesophagea of intensively raised pigs

Three groups of pigs (total 23 pigs) were fed ad libitum from weaning until market weight and these were compared with three groups of pigs (total 26 pigs) fed ad libitum until 12 weeks of age, and then fed on a restricted regimen (twice per day) until marketed. There was a significant difference in degree of ulceration between the two feeding regimens; those pigs fed ad libitum had the most severe gastric ulcers. The pigs fed ad libitum, had significantly less agonistic interactions per minute than did the restricted fed pigs. Growth rates of the pigs from weaning to market weight did not differ significantly in the two regimens, although the average pig on ad libitum feeding consumed 182.9 kg, and the restricted fed pigs ate 148.2 kg per pig.     

Perturbation of fuel homeostasis caused by overexpression of the glucose-6-phosphatase catalytic subunit in liver of normal rats

The terminal step in hepatic gluconeogenesis is catalyzed by glucose-6-phosphatase, an enzyme activity residing in the endoplasmic reticulum and consisting of a catalytic subunit (glucose-6-phosphatase (G6Pase)) and putative accessory transport proteins. We show that Zucker diabetic fatty rats (fa/fa), which are known to exhibit impaired suppression of hepatic glucose output, have 2.4-fold more glucose-6-phosphatase activity in liver than lean controls. To define the potential contribution of increased hepatic G6Pase to development of diabetes, we infused recombinant adenoviruses containing the G6Pase cDNA (AdCMV-G6Pase) or the beta-galactosidase gene into normal rats. Animals were studied by one of three protocols as follows: protocol 1, fed ad libitum for 7 days; protocol 2, fed ad libitum for 5 days, fasted overnight, and subjected to an oral glucose tolerance test; protocol 3, fed ad libitum for 4 days, fasted for 48 h, subjected to oral glucose tolerance test, and then allowed to refeed overnight. Hepatic glucose-6-phosphatase enzymatic activity was increased by 1.6-3-fold in microsomes isolated from AdCMV-G6Pase-treated animals in all three protocols, and the resultant metabolic profile was similar in each case. AdCMV-G6Pase-treated animals exhibited several of the abnormalities associated with early stage non-insulin-dependent diabetes mellitus, including glucose intolerance, hyperinsulinemia, decreased hepatic glycogen content, and increased peripheral (muscle) triglyceride stores. These animals also exhibited significant decreases in circulating free fatty acids and triglycerides, changes not normally associated with the disease. Our studies show that overexpression of G6Pase in liver is sufficient to perturb whole animal glucose and lipid homeostasis, possibly contributing to the development of metabolic abnormalities associated with diabetes.     

Determinants of coronary reserve in rats subjected to coronary artery ligation or aortic banding

OBJECTIVE: We investigated whether decreased coronary reserve in hearts after coronary artery ligation or in hearts from rats after aortic banding can be related to remodeling of resistance arteries. METHODS: Maximal coronary flow (absolute flow) and cardiac perfusion (flow corrected for heart weight) were determined in isolated, perfused rat hearts after adenosine or nitroprusside, at 3 and 8 weeks after coronary artery ligation or 4-5 weeks after aortic banding. Perivascular collagen and medial thickness of resistance arteries were determined by morphometry. RESULTS: maximal coronary flow of infarcted hearts had been restored to sham values at 3 weeks. Growth of cardiac muscle mass from 3 to 8 weeks exceeded the increase in maximal coronary flow, leading to a decreased perfusion at 8 weeks. A slight, transient increase in perivascular collagen, but no medial hypertrophy, was found after infarction. After aortic banding perivascular fibrosis and medial hypertrophy led to a decreased maximal coronary flow in both the hypertrophied left and the non-hypertrophied right ventricle. Consequently, perfusion of the left ventricle was most severely reduced. CONCLUSIONS: Reduced maximal perfusion after aortic banding is determined by both cardiac hypertrophy and vascular remodeling. In contrast, during infarction-induced remodeling, reduction of perfusion is not determined by vascular remodeling, but mainly by disproportional cardiac hypertrophy relative to vascular growth.     

Lactate dehydrogenase activity and isoform distribution in the rat pelvic ganglion: effects of diabetes and bladder outlet obstruction

We have previously shown that the intramural motor nerves in the rat bladder can function in anoxic conditions. The present study aims to explore the distribution and activity of lactate dehydrogenase (LDH), the key enzyme for ATP generation in anoxia. The activity and isoform distribution pattern of LDH was studied in pelvic ganglia from male and female rats. A histochemical investigation showed that the LDH activity was intense in the ganglion cells, and weak in the other tissue components (nerve bundles, connective tissue). The male pelvic ganglion weighted 55% more than the female pelvic ganglion, the enzyme activity per unit ganglion weight was 60% higher and the total LDH activity was 155% higher. The isoform distribution was similar, with M4 being dominant isoform, followed by M3H. Infravesical outlet obstruction in the female rat induced a threefold increase in ganglion weight, and the total LDH activity increased twofold. In this hypertrophic female ganglion a decreased relative amount of M4, and an increased amount of MH3, was found. Diabetes in the male rat had no effect on ganglion weight or its contents and isoform distribution of LDH.     

Cell proliferation and esophageal carcinogenesis in the zinc-deficient rat

Target cell proliferation was investigated throughout the development of esophageal cancer induced by N-nitroso-methylbenzylamine (NMBA) in weanling rats maintained on zinc-deficient or sufficient diets. Deficient rats were fed ad libitum, while zinc-sufficient rats were either pair-fed to the deficient animals or fed ad libitum. After 5 weeks, half of the animals in each dietary group were given six intragastric doses of NMBA (2 mg/kg; twice weekly). The remaining rats were untreated by carcinogen. At weeks 1, 2, 3, 4, 5, 7, 9 and 11 post first dose, esophageal cell proliferation was assessed in rats from each group by in vivo bromodeoxyuridine (BrDU) labeling followed by immunohistochemical detection of cells in S-phase. At 11 weeks, the tumor incidence was 100, 23 and 6%, respectively, in the zinc-deficient, zinc-sufficient, ad libitum and pair-fed groups. In vivo BrDU labeling revealed that in the NMBA-untreated groups, the labeling index (LI), the number of labeled cells, and the total number of cells per cross section of entire esophagi were significantly increased by zinc deficiency at all time points; LI was lowest in zinc-sufficient, pair-fed rats. During NMBA treatment (weeks 6, 7 and 8), increased cell proliferation occurred in both groups of zinc-sufficient esophagi but only during week 6 in the deficient ones. In the weeks following the cessation of NMBA treatment, zinc-deficient esophagi showed significantly increased LI and greater number of labeled cells than the carcinogen treated, zinc-sufficient pair-fed or ad libitum fed groups. On the other hand, NMBA-treated zinc-sufficient pair-fed rats showed lower LI and smaller number of labeled cells than their zinc-sufficient ad libitum counterparts. Most importantly, esophageal papillomas were found in two zinc-deficient animals that had received no NMBA treatment, after 10-11 weeks of experimental diet. These data support a direct relationship between cell proliferation and tumor incidence, and also provide evidence that zinc deficiency and its associated cell proliferation could be carcinogenic.     

Effect of hypodynamia on initial speed of lactate transport in skeletal muscle sarcolemmal vesicles in rats

Skeletal muscle sarcolemmal vesicles from control (C) and hindlimb suspended (S) rats were used to investigate the effect of unweighting on the lactate transporter activity. Sarcolemmal preparations were not different between the two groups. The efficiency of 4 weeks of hindlimb suspension was confirmed by a 40% decrease of citrate synthetase activity and a shift towards faster myosin isoforms in soleus muscle. The time course of 1 mM lactate uptake showed that the equilibrium was reached faster in group C (20 s) than in group S (40 s). The initial rate of 1 mM of lactate uptake decreased significantly (p < 0.05) after 4 weeks of hindlimb suspension. The initial rate of 50 mM lactate uptake did not differ significantly between the two groups. We conclude that 4 weeks of unweighting decreases significantly the skeletal muscle sarcolemmal lactate transport activity in rats. This result suggests that the level of physical activity probably plays a role on lactate transport regulation in muscle.     

Reduced levels of globular and asymmetric forms of acetylcholinesterase in rat left ventricle with pressure overload hypertrophy

The aim of the present work was to determine the effect of abdominal aortic stenosis on molecular forms of acetylcholinesterase (AChE) in rat heart. Pressure-overload, left ventricular hypertrophy was produced in male Sprague-Dawley rats by suprarenal abdominal aortic constriction. After two weeks the relative heart weight was increased over 20% compared to sham-surgical controls, mostly due to left ventricular enlargement. Aortic constriction reduced AChE activity per wet weight and per unit protein by 25-30% in the left ventricle and interventricular septum, but not in the other chambers. However, total AChE activity per chamber was normal in the left ventricle and interventricular septum, but was elevated in the atria. The molecular forms of AChE were separated in linear sucrose gradients and their specific activities were calculated from the resulting percent activities and total AChE activities. This data showed that although aortic constriction had no effect on ratios of the various forms, it did reduce the specific activities of globular and asymmetric forms in the left ventricle and interventricular septum. The reduced AChE activity suggests that slower rates of ACh hydrolysis occur in the left ventricle in pressure-overload hypertrophy.     

DNA sequence recognition by bis-linked netropsin and distamycin derivatives

The dihydropyridine receptor (DHPR), a voltage-gated L-type Ca2+ channel, and the Ca2+ release channel/ryanodine receptor isoform-1 (RyR1) are key molecules involved in skeletal muscle excitation-contraction coupling. We have reported age-related decreases in the level of DHPR expression in fast- and slow-twitch muscles from Fisher 344 cross Brown Norway (F344BNX) rats (Renganathan, Messi and Delbono, J. Membr. Biol. 157 (1997) 247-253). Based on these studies we postulate that excitation-contraction uncoupling is a basic mechanism for the decline in muscle force with aging (Delbono, Renganathan and Messi, Muscle Nerve Suppl. 5 (1997) S88-92). In the present study, we extended our studies to older ages and we intended to prevent or retard excitation-contraction uncoupling by restricting the caloric intake of the F344BNX rats from 16 weeks of age. Three age groups, 8-, 18-, and 33-month old caloric restricted rats, were compared with ad libitum fed animals. The number of DHPR and RyR1 and DHPR/RyR1 ratio (an index of the level of receptors uncoupling) in skeletal muscles of 8-month and 18-month rats was not significantly different in either ad libitum fed or caloric restricted rats. However, the age-related decrease in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in 33-month old ad libitum fed rats was absent in 33-month old caloric restricted rats. These results suggest that caloric restriction prevents age-related decreases in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in fast- and slow-twitch rat skeletal muscles.     

Effect of clenbuterol on growth and body composition during food restriction in rats

Clenbuterol was administered as a dietary admixture (4 mg/kg diet) to three groups of male Wistar rats (n = 8) housed individually in metabolism cages and fed for 15 d at 110, 160, and 235% (ad libitum) of estimated requirement for energy maintenance. Untreated groups at each level of energy intake were also included. There was no effect of clenbuterol on food intake in the ad libitum group, but the drug produced significant increases in body weight, feed efficiency, and carcass weight, dressing and protein content at all three levels of energy intake. This effect of clenbuterol was particularly noticeable in the restricted animals. Clenbuterol caused changes in body composition (increased percentage of water and protein, decreased percentage of fat) in the ad libitum rats but had no effect in the restricted groups. The reduction in the growth of the viscera caused by energy restriction was not affected by clenbuterol, apart from in the 110% restricted group, where the gastrointestinal tract was 26% heavier in the clenbuterol-treated rats. The results show that the growth anabolic actions of clenbuterol can be sustained and may be even more marked in rats fed restrictively than in those given ad libitum access to feed.     

The preferential mobilisation of C20 and C22 polyunsaturated fatty acids from the adipose tissue of the chick embryo: potential implications regarding the provision of essential fatty acids for neural development

The effects of 10 day clenbuterol administration on cardiac and skeletal muscle capillarities were studied, particularly in terms of the distribution of arteriolar and venular capillaries and their capillary density, in young (10-week-old) and middle-aged (37-week-old) male Wistar rats. Rats of the treated groups were fed a diet containing 2 mg kg-1 clenbuterol hydrochloride. In both young and middle aged rats, clenbuterol treatment increased the body wt and the weights of the heart and hindlimb muscles. The mean fibre cross-sectional area was significantly increased after the treatment in the left ventricle, soleus, plantaris and both deep and superficial portions of gastrocnemius (P < 0.01). In the left ventricle, the total capillary density and the density of venular capillaries were decreased after the treatment in both young (9 and 13%, respectively) and middle-aged rats (10 and 11%, respectively). A decrease in total capillary density was also observed in all skeletal muscles examined. In both young and middle-aged rats, the capillary-to-fibre (C:F) ratio and the proportion of each capillary did not change after the treatment in both the left ventricle and skeletal muscles. Clenbuterol significantly decreased the activity of succinate dehydrogenase in all skeletal muscles examined (P < 0.01). These results suggest that clenbuterol increased the diffusion distance for oxygen in the left ventricle and skeletal muscles. These changes may reduce the oxygen supply to tissues and increase muscle fatigability.     

Ornithine decarboxylase activity in muscle, liver, and intestinal tissue of turkeys during a short-term feed withdrawal and following refeeding

The activity of ornithine decarboxylase (ODC), an enzyme associated with cellular growth and protein synthesis, was examined in breast muscle, liver, and intestinal tissues of turkeys during a short-term period of feed withdrawal (FW) and following refeeding. Turkeys from a randombred control line were reared under standard management practices to 3 wk of age in battery brooders. Feed was then withdrawn from FW birds for a 48-h period, after which feed was consumed ad libitum. Control birds consumed feed ad libitum throughout the test period. Tissues were collected from 12 birds per treatment following 24 and 48 h of FW and at 6, 12, 24, and 48 h following refeeding for later determination of tissue ODC activity. Activity of ODC was greater in tissue from the small intestine than in liver tissue and both had greater activity than that exhibited by breast muscle. Short-term FW and refeeding produced differential responses in ODC activity of the three tissues examined. Feed withdrawal resulted in a reduction of ODC activity in intestinal tissue, whereas activity was unaffected for liver or breast muscle tissues. Compensatory increases in ODC activity were observed in liver and intestinal tissues; however, the increase was both more rapid and transitory in small intestine than in liver tissue. The ODC activity in breast muscle was largely unaffected by short-term FW and refeeding. Patterns of ODC activity in liver during FW and refeeding closely resembled patterns observed for absolute and relative liver weight. Thus, the results of the present experiment demonstrate that short-term FW and refeeding influence underlying growth mechanisms of supply organs, such as hepatic and intestinal tissue, in addition to affecting overall growth and muscle development.     

Quantitative determination of amino acid levels in neutral and glucosamine-containing carbohydrate polymers

OBJECTIVE: To determine whether local cardiac angiotensin converting enzyme (ACE) expression is upregulated during the development of hypoxia-induced right ventricular hypertrophy. METHODS: ACE activity was measured in membrane preparations from the right ventricle and left ventricle plus septum in normoxic rats and animals exposed to chronic hypoxia for 8 and 14 days. Local cardiac ACE expression was studied by immunohistochemistry using a monoclonal antibody to ACE (9B9). RESULTS: In the normal rat heart, ACE expression was confined to vascular endothelium, the valvular endocardium, and localized regions of parietal endocardium. We found that the development of pulmonary hypertension and right ventricular hypertrophy were associated with 2.6- and 3.4-fold increases in membrane-bound right ventricular ACE activity by 8 and 14 days of hypoxia, respectively. Right ventricular ACE activity was positively correlated with the degree of right ventricular hypertrophy (r = 0.83, P < 0.001). In contrast, left ventricular plus septal ACE activity was significantly reduced by approximately 40 and 60% by 8 and 14 days of hypoxia, respectively, compared to controls. In the right ventricle of chronically hypoxic rats, immunohistochemistry demonstrated increased ACE expression in areas of myocardial fibrosis. Interestingly, increased ACE expression was noted in the right ventricular epicardium in chronically hypoxic rats. In the free wall of the left ventricle there was a significant reduction in the number of myocardial capillaries which expressed ACE in chronically hypoxic rats. CONCLUSION: Chronic hypoxia has a differential effect on left and right ventricular ACE activity and that the sites of altered ACE expression are highly localized. We speculate that locally increased right ventricular ACE activity and expression may play a role in the pathogenesis of right ventricular hypertrophy secondary to hypoxic pulmonary hypertension.