Excitatory amino acid receptors and puberty

Glutamate is an important excitatory signal in the hypothalamus for the steroid-mediated preovulatory gonadotropin surge. Steroids may exert this action by regulating glutamate receptor levels or glutamate release, or both. Work in our laboratory found no changes in NMDA and kainate receptor binding in the hypothalamus of castrated or castrated plus steroid-replaced male and female rats. Likewise, we found that NMDA and kainate binding did not change over the onset of puberty in the female rat. A competitive quantitative RT-PCR assay using exogenous internal standards was used to measure NMDAR1, GluR1, and beta-actin mRNAs levels. NMDAR1 and GluR1 expression was examined in the preoptic hypothalamic area and in the medial basal hypothalamus at Postnatal Days 10, 15, 20, 25, 30, 32, 34, 36, 40, and 63. A transient increase in GluR1 mRNA levels in the preoptic hypothalamic area was observed on Day 20, with all other time points showing comparable levels. NMDAR1 levels in the POA and medial basal hypothalamus did not change significantly at any of the time points; in contrast, however, AMPA receptor binding levels were increased in the hypothalamus at the time of puberty in the female rat. Thus, in addition to the previously reported elevation of glutamate release rates in the hypothalamus at the time of puberty, AMPA receptors may also be elevated and play a role in mediating glutamate regulatory effects on the timing of puberty in the female rat.     

Effect of ascorbic acid deficiency on primary and reparative dentinogenesis in non-ascorbate-synthesizing ODS rats

Ascorbic acid is essential to the biosynthesis of collagen, the major organic matrix component of dentine. The ODS rat is a mutant strain of Wistar rat characterized by hereditary lack of L-gulono-gamma-lactone oxidase and thus is unable to synthesize ascorbic acid. ODS rats were given an ascorbic acid-free diet to investigate how ascorbic acid deficiency affects dentine formation in vivo. Histomorphometric analysis on their growing molars and incisors showed a significant reduction in both size and mineral apposition rate of dentine, as revealed by contact microradiography and fluorescent time-marking, respectively. A similar reduction in bone formation was simultaneously demonstrated in the mandible, confirming the previously reported osteopathic effects of ascorbic acid deficiency. When pulp inflammation was induced in lower first molars by making unsealed pulp exposures, specimens from control animals showed continuous deposition of an osteodentine-like tissue in the radicular pulp chamber; this type of mineralized tissue formation was greatly reduced in ascorbic acid-deprived animals. These results indicate that ascorbic acid deficiency hampers dentine formation under both physiological and pathological conditions of the dentine/pulp complex. ODS rats could be useful in investigating in vivo effects of ascorbic acid deficiency on the formation of dentine and other dental mineralized tissues.     

Time after feeding and dietary arginine deficiency alter splanchnic and hepatic amino acid flux in rats

The effect of an arginine-deficient diet containing 3.4% glutamate on net flux of amino acids across the portal-drained viscera and liver was studied in rats at 0, 1 or 2 h after a meal and compared with that in arginine-fed controls. Net portal-drained viscera flux for most amino acids was greater in the fed state compared with the postabsorptive state except for glycine and cystine, which did not change, and methionine, which declined. Net amino acid recovery in portal blood 2 h after feeding compared with amounts consumed was highest for alanine (17.3%); recovery of other amino acids ranged from 5.6 to 15.3%. No net portal-drained viscera recovery of consumed cystine was observed. For the branched-chain and aromatic amino acids, methionine, threonine, histidine and lysine, net hepatic uptake was nearly equal to net portal-drained viscera absorption (range 77-127% of portal-drained viscera flux). Correlation coefficients between net hepatic and portal-drained viscera fluxes for leucine, valine, isoleucine, methionine and phenylalanine were 0.84 to 0.93. Postabsorptive hepatic extraction for most amino acids was zero, but after a meal, ranged from 13.3 to 22.9% for the branched-chain and aromatic amino acids. Net hepatic production of ornithine and proline occurred in arginine-fed control rats. This value was near zero for ornithine in rats fed the arginine-deficient diet. Models of interorgan amino acid metabolism in the food-deprived and fed state are presented.     

Effect of varying amino acid levels on protein metabolism in nephrotic rats during total parenteral nutrition

In an attempt to determine appropriate diet in nephrotic syndrome, nephrotic rats, induced by puromycin aminonucleoside, were nourished by total parenteral nutrition fluid containing the same energy, but three different levels (1.65, 3.3, and 6.6%) of amino acids for 7 d. The fractional rate of total protein synthesis in the liver was determined by injecting a flooding dose of [3H]phenylalanine. The proportion of newly synthesized proteins retained and exported by the liver was estimated by injecting a tracer dose of [14C]leucine and then measuring the protein radioactivity remaining in the liver and present in the plasma after secretion was completed. Nephrotic animals synthesized more protein than control animals. Although the absolute synthesis rates of total protein in liver were increased with increasing amino acid administration, the absolute rates of synthesis of albumin were higher in the 3.3% group than in the other groups in nephrotic rats. However, kidney protein synthesis in nephrotic rats was higher in the 1.65% group than in the 3.3% group. Interestingly, the 3.3% group revealed the smallest urinary excretion of total protein and albumin. In addition, in the 3.3% group, plasma concentrations of total protein and albumin were higher, and plasma concentrations of total cholesterol and triglyceride were lower than in other groups. It was concluded that the 3.3% group, corresponding to a normal protein diet, has the greatest salutary effect on urinary protein excretion, followed by protein and lipid metabolism, in nephrotic rats. Not only protein intake but also the energy:protein ratio are important for diet therapy in nephrotic animals. The technique of total parenteral nutrition may be useful in defining the factors involved in glomerular permeability or permselectivity and intracellular protein metabolism.     

Influence of undernutrition on the lenticular epithelium in neonatal and weanling albino rats

Protein malnutrition, a morbid nutritional disorder of the developing countries, is known to interfere with cell replication. The influence of undernutrition on the lens which represents a unique combination of static, conditional renewal and constant cell renewal systems is not completely understood. In the present investigation it has been shown that undernutrition results in poor synthetic and proliferative activities of lenticular epithelium in neonatal and post-weaning rats. It is supposed that protein deficiency per se may not produce clinical diseases, but may increase the susceptibility of lenses to exogenous and/or endogenous insults.     

Epidermal growth factor (EGF) increases the renal amino acid transport capacity in amino acid loaded rats

In anaesthetized adult female rats, the influence of epidermal growth factor (EGF) on renal amino acid handling was investigated in glutamine, arginine (both 50 mg/100 g b.wt. per hour), or alanine (90 mg/100 g b.wt. per hour) loaded animals. Continuous infusions of the three amino acids were followed by an increase in the fractional excretion (FE) of the administered amino acids as well as of the other endogenous amino acids. Under load conditions (alanine, arginine or glutamine), EGF pretreatment (8 micrograms/100 g b.wt. subcutaneously for 8 days, twice daily 8 a.m. and 4 p.m.) was followed by a stimulation of renal amino acid reabsorption. The increase in the fractional excretion of the administered amino acids was significantly lower than in non-EGF-treated rats. These changes in amino acid transport were connected with a significant reduction of GFR after EGF pretreatment (0.96 +/- 0.10 vs. 0.62 +/- 0.07 ml/min x 100 g b.wt.) and a distinct increase in sodium excretion (2.98 +/- 0.55 vs. 4.97 +/- 0.71 muval/100 g b.wt. x 20 min). After loading with p-aminohippurate (PAH; 200 mg/100 g b.wt.), PAH excretion in EGF rats was increased by about 20%, whereas urinary protein excretion was lower in EGF pretreated rats (control: 0.45 +/- 0.04 vs. EGF: 0.18 +/- 0.03 mg/100 g b.wt. x 20 min). The PAH load reduced amino acid reabsorption as a sign of overloading of renal tubular transport capacity, but in EGF pretreated animals the amino acid excretion was only slightly increased under these conditions. Furthermore, EGF pretreatment depressed normal kidney weight gain significantly (874 +/- 18 vs. 775 +/- 32 mg/100 g b.wt.). EGF can improve the renal tubular transport capacity, but, compared to well-known stimulators of renal transport like dexamethasone or triiodothyronine, its effect is only of a moderate degree.     

The hepato-renal syndrome: renal amino acid transport in bile duct ligated rats (DL)--influence of treatment with triiodothyronine or dexamethasone on renal amino acid handling in amino acid loaded rats

The influence of triiodothyronine or dexamethasone on renal amino acid handling was investigated in anaesthetized, bile duct-ligated (DL) adult female rats. 3 days after DL, glomerular filtration rate (GFR) was unchanged whereas urine flow was decreased. Plasma concentrations of 5 out of 16 amino acids were significantly enhanced after DL. On the other hand, the fractional excretion (FE) of 11 out of 16 amino acids was significantly reduced as a sign of improved reabsorption capacity. Bolus injections of leucine (20 mg/100 g b.wt.), glutamine (45 mg/100 g b.wt.), or taurine (45 mg/100 g b.wt.) were followed by a temporary increase in the FE of the administered amino acids as well of the endogenous amino acids which were not administered. This phenomenon was more pronounced in DL than in control rats. Under load conditions, dexamethasone (60 microg/100 g b.wt.) or triiodothyronine (20 microg/100 g b.wt.) treatment for 3 days, i.p. once daily, was followed by a stimulation of renal amino acid reabsorption in DL rats. The increase in fractional amino acid excretion after amino acid load was significantly lower than in untreated rats. This effect was also more pronounced in DL rats.     

Effect of increasing dietary threonine intakes on amino acid metabolism of the central nervous system and peripheral tissues in growing rats

Although clinical rating scales and simple timed tests of motor function are widely used to assess motor response to therapy, gait analysis may provide an alternative measure of this response. We studied 15 patients with PD complicated by motor fluctuations, first to determine changes in temporal and spatial gait parameters following levodopa, secondly to assess the stability of repeated gait measures and timed tests in "off" and "on" states, and thirdly to determine the use of gait analysis in the assessment of the dopaminergic response. Gait analysis (velocity, stride length, cadence, and double limb support), clinical rating scales (modified Webster scale and Hoehn and Yahr stage), and timed tests of motor function (hand tapping and stand-walk-sit time) were performed before ("off") and after ("on") a levodopa challenge. Stride length and gait velocity increased following medication whereas cadence and double limb support did not. Most gait measures and the stand-walk-sit time were stable over three consecutive trials in both "off" and "on" states. Of the gait measures, only cadence in the "off" state changed significantly whereas the tapping count improved with repeated trials in both "off" and "on" states. Changes in stride length, gait velocity, and tapping count following levodopa correlated with changes in clinical rating scales following treatment. Measurement of gait parameters provides a reliable, objective alternative to rating scales and timed tests in assessing the dopaminergic response in patients with PD and motor fluctuations.     

Patterns of growth deficiency in rats exposed in utero to undernutrition, ethanol, or the neuroteratogen methylazoxymethanol (MAM)

Children and experimental animals exposed to ethanol (EtOH) in utero commonly have low birthweights, and many remain small at maturity. Low body weight or small stature in adulthood may reflect an inability to recover from in utero growth retardation, or it may reflect a separate, postnatal growth deficiency. In this study, daily body weights (postnatal days 1 to 60) were compared among the offspring of the following groups of Long Evans rats: dams fed liquid diet containing 35% EtOH-derived calories; their pair-fed and chow-fed controls; and dams exposed to methylazoxymethanol (MAM) in two previous studies, in which offspring exhibited reduced numbers of growth hormone releasing factor (GRF) neurons. All treatments produced a number of offspring with weight deficits beginning after birth and persisting into maturity. Three distinct patterns of growth deficiency were observed: (1) weight loss relative to controls in the first weeks of life, seen in offspring exposed to EtOH, pair feeding, or MAM on gestation day 13 (G13); (2) a delay in the onset of the prepubertal growth spurt, seen in all EtOH-exposed offspring and in G13 MAM-exposed dwarfs; and (3) failure to sustain the prepubertal growth spurt, seen only after exposure to MAM on G14. The results of this study support the view that prenatal EtOH exposure is capable of affecting postnatal growth specifically; moreover, the pattern of growth deficiency seen in EtOH-exposed offspring was distinct from that of the undernourished offspring of pair-fed dams.     

Microbial amino acid synthesis and utilization in rats: the role of coprophagy

The Substance Abuse Monitor is a comprehensive community-based addictions information system that forms the basis for an innovative approach to community needs assessment. This paper describes: (1) the conditions that created the need and support for the monitoring system; (2) the community development strategy that was employed to secure the commitment of agencies to the project; (3) the specifics of the needs assessment procedures; (4) some of the theoretical, clinical and methodological issues on which the procedures are based; (5) the practical applications of the system; and (6) the limitations of the system. The establishment of the database was based upon the idea that there would be benefits at several levels: the community, the participating agencies, the individual counsellors and the clients. The realization of these benefits requires that the information collected and the results generated must be capable of addressing specific service delivery problems with practical and tangible solutions. It is argued that the project has been successful in generating such solutions, and has considerable potential as an ongoing needs assessment tool.     

Small changes in essential amino acid concentrations alter diet selection in amino acid-deficient rats

A new plate designed specifically to address complex wrist pathology was used for the internal fixation of 22 complex fractures of the distal radius in 22 patients in a prospective multicenter trial. The majority of fractures were group C2- and C3-type fractures according to the Comprehensive Classification of Fractures. No plate failures, loss of reduction, nonunions, or infections occurred. Within the average follow-up time of 14 months, the functional results (including an average motion of 76% and an average grip strength of 56% of the contralateral side) were comparable to those reported for similar fractures in previous investigations. Five patients had irritation of the tendons in the second dorsal compartment. This trial serves both as a verification of the safety and efficacy of this distal radius plate as well as a demonstration of its utility in the treatment of complex fractures of the distal radius.     

Effects of methionine sulphoximine treatment on renal amino acid and ammonia metabolism in rats

Renal glutamine metabolism in relation to ammoniagenesis has been extensively studied during chronic metabolic acidosis, when arterial glutamine levels are reduced. However, little is known about the effects of reduced glutamine delivery on renal glutamine and ammonia metabolism at physiological systemic pH values. Therefore, a model of decreased arterial glutamine concentrations at normal pH values was developed using methionine sulphoximine (MSO). Renal glutamine and ammonia metabolism was measured by determining fluxes and intracellular concentrations after an overnight fast in ether anaesthetized normal rats, MSO-treated rats and their pair-fed controls. Moreover, fluxes and intracellular concentrations of several other amino acids were determined concomitantly. After 2 and 4 days of MSO treatment, arterial glutamine concentrations were reduced to 55%, while arterial ammonia concentrations increased by 70%. Kidney glutamine uptake reduced, but systemic pH was unchanged. Fractional extraction of glutamine remained unchanged, suggesting that also in vivo net uptake of glutamine by the kidney at subnormal levels is related to arterial glutamine concentrations. As a result, at day 2 but not at day 4, the kidney reduced the net release of ammonia into the renal vein and thus reduced net renal ammonia addition to body ammonia pools. Therefore at day 2, the kidney seems to play an important role in adaptation to both hyperammonaemia and hypoglutaminaemia.     

Physiological perspectives on leptin as a regulator of reproduction: role in timing puberty

Effects of tumor stimulator cell modification by infection with Newcastle Disease Virus (NDV) are described as analysed in vitro in mixed lymphocyte tumor cell cultures (MLTC). Direct antitumor effects were seen with human melanoma or colon-carcinoma cells in a dose- and time-dependent manner when using live but not UV inactivated virus. When T cell stimulation was measured by [3H]-thymidine uptake, NDV infected tumor stimulator cells did not show an augmentation but rather an inhibitory effect in comparison to non-infected stimulator cells. Virus infected tumor stimulator cells were, however, capable of augmenting the induction of tumor specific cytotoxic T cells in MLTC-CML assays when using murine ESb lymphoma immune cells and syngeneic NDV modified ESb cells as stimulators. A CML stimulatory effect was also shown for NDV modified third party cells and thereof derived conditioned medium. These effects are most likely explained by interferon- which is induced in tumor cells by NDV infection and by interferon-á which is induced in responder cells when stimulated with NDV infected stimulator cells.     

Pre-exercise branched-chain amino acid administration increases endurance performance in rats

This study investigated the effects of pre-exercise branched-chain amino acid (BCAA) administration on blood ammonia levels and on time to exhaustion during treadmill exercise in rats. Adult female Wistar rats were trained on a motor driven treadmill. After a 24-h fast, rats were injected intraperitoneally (i.p.) with 1 mL of placebo or BCAA (30 mg), 5 min before performing 30 min of submaximal exercise (N = 18) or running to exhaustion (N = 12). In both cases, rats were sacrificed immediately following exercise, and blood was collected for the measurement of glucose, nonesterified fatty acid (NEFA), lactic acid, BCAA, ammonia, and free-tryptophan (free-TRP) levels. Control values were obtained from sedentary rats that were subjected to identical treatments and procedures (N = 30). Plasma BCAA levels increased threefold within 5 min after BCAA administration. Mean run time to exhaustion was significantly longer (P < 0.01) after BCAA administration (99 +/- 9 min) compared with placebo (76 +/- 4 min). During exercise, blood ammonia levels were significantly higher (P < 0.01) in the BCAA treated compared with those in the placebo treated rats both in the 30-min exercise bout (113 +/- 25 mumol.L-1 (BCAA) vs 89 +/- 16 mumol.L-1) and following exercise to exhaustion (186 +/- 44 mumol.L-1 (BCAA) vs 123 +/- 19 mumol.L-1). These data demonstrate that BCAA administration in rats results in enhanced endurance performance and an increase in blood ammonia during exercise.     

Impact of puberty on family relations: Effects of pubertal status and pubertal timing.

This investigation examines the impact of pubertal status and pubertal timing, independent of each other and of chronological age, on the family relationships of adolescent boys and girls. The sample is composed of 204 families with a firstborn child between the ages of 10 and 15. Measures included adolescent and parental reports of closeness, conflict, and autonomy as well as assessments of each adolescent's pubertal status and pubertal timing (early, on time, or late maturing). Findings indicate that (a) pubertal maturation is associated with increased emotional distance between youngsters and their parents; (b) pubertal maturation (among girls) and early pubertal maturation (among boys) increase conflict between adolescents and their mothers, but not necessarily fathers; and (c) pubertal maturation, and especially late maturation, may be accompanied by increased behavioral autonomy for the adolescent. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Influence of chronic undernutrition on oxygen consumption of rats during exercise

The current study was undertaken to examine the effects of dithiothreitol (DDT), a sulfhydryl-reducing agent and heavy metal chelator, on the course of heavy metal-induced acute renal failure in the rat. Groups of rats in metabolic cages received uranyl nitrate (UN) alone, UN plus DTT, mercuric chloride (HgCl2) alone, and HgCl2 plus DTT. UN injected alone produced azotemia, decreased creatinine clearance, and rising fractional sodium excretion over the 48 hr of study. These effects of UN on renal function were not observed when DTT was administered 30 min after UN injection. Qualitatively similar results were obtained with HgCl2-induced acute renal failure. Groups of rats were killed at 6 hr after UN plus DTT, HgCl2 alone, or HgCl2 plus DTT; and determinations of plasma renin activity (PRA) and renin activities of the superficial and deep juxtaglomerular apparatus (JGA) were performed. PRA's and JGA renins were increased in animals receiving either UN or HgCl2 alone, but not in the rats receiving both DTT and UN or HgCl2. The effect of DTT on distribution of 203Hg was also examined. Treatment with DTT did not alter the renal accumulation of 203Hg, suggesting that this agent does not act by limiting renal exposure to the heavy metals. Thus, DTT ameliorates the course of heavy metal-induced ARF, and this effect is associated with prevention of heavy metal-induced alterations in sodium excretion and renin-angiotensin system activity.     

Effect of acute heat stress on amino acid digestibility in laying hens

An experiment was conducted to determine the effect of acute heat stress exposure on amino acid digestibility in laying hens. A total of 30 commercial laying hens were singly housed in an environmentally controlled facility, fed a standard laying ration, and exposed to a constant thermoneutral temperature (21 C) for 12 d. The hens were then randomly fed one of three diets (10 hens per diet) and exposed to three consecutive temperature periods (8 d each), which consisted of: 1) a constant 21 C temperature, 2) a cycling temperature of 35 C for 12 h and 29 C for 12 h, and 3) a constant 21 C temperature. The three isonitrogenous (18% CP) diets fed were: 1) a corn-soybean meal diet, 2) a corn-soybean meal diet containing 15% meat and bone meal, and 3) a corn-soybean meal diet containing 5% alfalfa meal and 20% wheat bran. Excreta were collected from all hens during the last 4 d of each temperature period and apparent amino acid digestibility was determined. There was a significant diet effect (P < 0.05) on amino acid digestibility. Digestibility of amino acids in Diet 2 (corn-soybean meal/meat and bone meal) was higher (P < 0.05) than in the other two diets. In addition, digestibility of amino acids in Diet 3 (corn-soybean meal/alfalfa meal/wheat bran) was significantly lower (P < 0.05) than in Diets 1 or 2. Heat stress generally had no significant effect on amino acid digestibility except for His and Lys digestibility. Histidine digestibility was higher during the heat stress period than during the initial and recovery thermoneutral periods, whereas Lys digestibility was higher during the heat stress period than during the initial thermoneutral period. These results indicated that acute heat stress (8 d) had no adverse effects on dietary amino acid digestibility in laying hens.     

Effect of amino acid supplementation on whole-body protein turnover in Holstein steers

We used the [15N]glycine single-dose urea end-product technique to measure whole-body protein turnover in six Holstein steers (250 +/- 18 kg). Steers were implanted with Revalor-S and continuously infused abomasally with water (4 L/d) or amino acids (AA; in 4 L/d water) in a crossover experiment (two 14-d periods). The AA infusion contained the following (g/d): lysine (5.3), methionine (3.3), threonine (3.2), tryptophan (1.0), histidine (2.1), and arginine (5.5). Steers were fed a diet containing 85% rolled corn, 10% prairie hay, and 1.1% urea (DM basis) at 2.16% of body weight. Nitrogen retention tended (P = .15) to increase with AA infusion, from 27.9 to 32.9 g N/d. Amino acid infusion numerically increased whole-body protein turnover from 168.6 to 183.2 g N/d, protein synthesis from 152.6 to 169.3 g N/ d, and protein degradation from 124.7 to 136.4 g N/d. Enhanced protein accretion may have resulted from a larger increase in protein synthesis than in degradation. The tendency for increased N retention is interpreted to suggest that the implanted, lightweight Holstein steers fed a corn-urea diet in our study were able to respond to AA supplementation, suggesting that at least one of the infused AA was limiting in the basal diet. Protein turnover data suggest that cattle, like other animals, may increase protein synthesis and protein degradation in response to supplementation with limiting AA. The [15N]glycine single-dose urea end-product technique for measuring whole-body protein turnover in cattle may be useful.