Effects of dietary crude protein and tannic acid on nitrogen excretion, urinary nitrogenous composition and urine nitrous oxide emissions in beef cattle.

Two consecutive trials were carried out to study the effects of dietary crude protein (CP) and tannic acid (TA) on nitrogen (N) metabolism of beef cattle and consequently, the N2 O emissions from the urine of cattle. In Trial I, eight growing castrated cattle were used as the experimental animals. Two levels of dietary CP (110.6 and 135.7 g/kg dry matter [DM]) and two levels of TA (0 and 16.9 g/kg DM) were allocated in a replicated 2 × 2 crossover design. In Trial II, the N2 O emissions from the urine of cattle collected from Trial I were determined using the static incubation technique. An interaction between dietary CP and TA on the urinary N excretion (p < .05) was found but not on the N2 O-N emission of cattle urine. Increasing dietary CP level from 110.6 g/kg DM to 135.7 g/kg DM increased the total N excretion (p < .001), the N retention (p < .05) and the ratio of urinary urea-N/urinary N (p < .01), did not affect the N use efficiency (NUE; p > .05) and shifted the N excretion from faeces to urine. Increasing the dietary CP level increased the N2 O-N emission of cattle urine. Dietary addition of TA decreased the urinary excretions of urea (p < .001) and shifted the N excretion from urine to faeces, did not affect the NUE of beef cattle (p > .10), and decreased the N2 O-N emission of cattle urine. Pyrogallol and resorcinol of the TA metabolites were detected in urine with dietary addition of TA. Feeding beef cattle with relatively low CP level and adding TA in rations are effective approaches to mitigate the N2 O-N emissions from cattle urine.

Nitrogenous and phosphorus excretions in juvenile silver catfish (Rhamdia quelen) exposed to different water hardness, humic acid, and pH levels.

This study examined ammonia, urea, creatinine, protein, nitrite, nitrate, and phosphorus (P) excretion at different water hardness, humic acid, or pH levels in silver catfish (Rhamdia quelen) juveniles. The fish were exposed to different levels of water hardness (4, 24, 50, or 100 mg L(-1) CaCO3), humic acid (0, 2.5, or 5.0 mg L(-1)), or pH (5.0, 6.0, 7.0, 8.0, or 9.0) for 10 days. The overall measured nitrogen excretions were 88.1% (244-423 µmol kg(-1 )h(-1)) for ammonia, 10.9% (30-52 µmol kg(-1 )h(-1)) for creatinine, 0.02% (0.05-0.08 µmol kg(-1 )h(-1)) for protein, 0.001 % (0.002-0.004 µmol kg(-1 )h(-1)) for urea, 0.5% (0.64-3.6 µmol kg(-1 )h(-1)) for nitrite, and 0.5% (0.0-6.9 µmol kg(-1 )h(-1)) for nitrate, and these proportions were not affected by water hardness or humic acid levels. The overall P excretion in R. quelen was 0.14-2.97 µmol kg(-1) h(-1). Ammonia excretion in R. quelen usually was significantly higher in the first 12 h after feeding, and no clear effect of water hardness, humic acid levels, and pH on this daily pattern of ammonia excretion could be observed. Water hardness only affected the ammonia and P excretion of R. quelen juveniles in the initial and fifth days after transfer, respectively. The exposure of this species to humic acid increased ammonia excretion after 10 days of exposure but did not affect P excretion. An increase in pH decreased ammonia and increased creatinine excretion but did not change P excretion in R. quelen. Therefore, when there is any change on humic acid levels or pH in the culture of this species, nitrogenous compounds must be monitored because their excretion rates are variable. On the other hand, P excretion rates determined in the present study are applicable to a wide range of fish culture conditions.

Effects of malate on diet digestibility, microbial protein synthesis, plasma metabolites, and performance of growing lambs fed a high-concentrate diet.

The objective of this study was to evaluate the effects of malate supplementation on growth rate, feed efficiency, and diet digestibility in growing lambs. Twenty-four Merino lambs with a mean BW of 15.3 +/- 0.22 kg were divided into 3 homogenous groups. Each group was randomly allocated to 1 of 3 malate (16% disodium malate:84% calcium malate) levels: 0 (control), 4 (MAL-4), or 8 (MAL-8) g/kg of concentrate. Lambs were fed concentrate and barley straw ad libitum for 35 d. After a 20-d period, diet digestibility was determined, and microbial N flow at the duodenum was estimated from the urinary excretion of purine derivatives. Blood samples were taken on d 0, 20, and 35. On d 35, lambs were slaughtered and ruminal fluid samples were collected. There were no effects (P = 0.18 to P = 0.95) of malate on concentrate or straw intake, ADG, carcass yield, and apparent digestibility of OM, CP, NDF, or ADF. Malate supplementation did not influence (P = 0.80) the daily urinary excretion of total purine derivatives, and therefore there were no treatment effects (P = 0.77) on estimated microbial N flow at the duodenum. No differences (P > 0.05) among treatments were observed for plasma concentrations of glucose, cholesterol, triglycerides, urea-N, lactate, or VFA, but malate addition increased (P = 0.003) the molar proportion of butyrate in ruminal fluid (4.29, 6.14, and 5.45% of total VFA for control, MAL-4 and MAL-8, respectively). The use of malate as a feed additive under the conditions of the current study did not influence diet intake or digestion, and consequently did not improve lamb performance.

Renal protective role of bradykinin B1 receptor in stroke-prone spontaneously hypertensive rats.

The kallikrein-kinin system plays important roles in blood pressure regulation, metabolism of electrolytes and organ protection. Although the bradykinin B2 receptor (B2R) has been reported to be involved in most of these effects, a role of the bradykinin B1 receptor (B1R) has also been noted recently. The aim of this study was to determine the role of renal B1R in stroke-prone spontaneously hypertensive rats (SHR-SP). Sixteen-week-old SHR-SP and Wistar Kyoto rats (WKY) as a control were used in the experiments. A high level of B1R mRNA was detected in SHR-SP, while the expression in WKY was almost undetectable. Immunohistochemistry revealed a B1R protein in the renal tubules and glomeruli in SHR-SP. The acute injection of a B1 R agonist into SHR-SP increased urinary NOx excretion to a level up to 5-fold higher than that in the SHR-SP treated with vehicle. The infusion of B1 R antagonist for 4 weeks resulted in a significant elevation of blood pressure and urinary albumin excretion and a decrease in urinary NOx excretion in SHR-SP. The administration of B1 R antagonist resulted in renal interstitial and glomerular fibrosis in SHR-SP. Moreover, the expressions of transforming growth factor (TGF) beta1 protein and collagen III mRNA in SHR-SP treated with B1R antagonist were significantly higher than those of SHR-SP treated with a vehicle. The expression and phosphorylation of extracellular signal-regulated protein kinase (ERK) and p38, but not c-Jun N-terminal kinase (JNK), were significantly increased in the SHR-SP treated with B1R antagonist. These results indicated that renal B1R might be over-expressed in a high blood pressure condition, and that this upregulated B1 R may play an important role in renal protection by inhibiting renal fibrosis via an increase of NO production and a suppression of TGFbeta1 expression and mitogen-activated protein kinase (ERK and p38) phosphorylation.

The detection and identification of quaternary nitrogen muscle relaxants in biological fluids and tissues by ion-trap LC-ESI-MS.

Quaternary nitrogen muscle relaxants pancuronium, rocuronium, vecuronium, gallamine, suxamethonium, mivacurium, and atracurium and its metabolites were extracted from whole blood and other biological fluids and tissues by using a solid-phase extraction procedure. The extracts were examined by using high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). The drugs were separated on a ODS column in a gradient of ammonium acetate buffer (pH 5.0) and acetonitrile. Full-scan mass spectra of the compounds showed molecular ions, and MS-MS spectra showed fragments typical of the particular compounds. LC-ESI-MS allowed an unequivocal differentiation of all muscle relaxants involved. The method was applied in a case of rocuronium and suxamethonium administration in a Caesarian section and in a case of intoxication by pancuronium injection. In both cases, the administered drugs could be detected and identified in the supplied samples.

Factors affecting the first recurrence of noncardioembolic ischemic stroke.

Studies of the factors affecting the first recurrence of ischemic stroke have reported inconsistent findings. Types of initial stroke and the racial differences in study samples are among the explanations that may account for this inconsistency. The aims of this study were to estimate the cumulative recurrence rates of noncardioembolic ischemic stroke and identify the factors that influence the first recurrence of noncardioembolic ischemic stroke in the Taiwanese Chinese population. Four hundred and sixty-six patients with noncardioembolic ischemic stroke from thirteen hospitals in Taiwan were followed up in this study to ascertain first recurrence of noncardioembolic ischemic stroke between October 1992 and April 1995. The Kaplan-Meier method was used to estimate the cumulative recurrence rate. The Cox regression model was used to ascertain the significant factors affecting the first recurrence of noncardioembolic ischemic stroke. The overall cumulative recurrence rate was 10.5% (49/466) from the follow-up period of 30 months. After adjustment for age, sex, treatment modes, and variables pertinent to blood pressure, the site of brain lesion remained a significant factor. The relative risk of first recurrence for the basal ganglion vs. the region of middle cerebral artery was 3.06 (95% CI: 1.29-7.26). The brain lesion site was demonstrated to be an independent predictor of risk for the first recurrence of noncardioembolic ischemic stroke among the Taiwanese Chinese population. Whether this finding was also seen in other populations should be corroborated in future research.

Dietary prescription based on estimated nitrogen balance during peritoneal dialysis.

Protein and energy requirements of children on automated peritoneal dialysis (APD) have still not been sufficiently well defined, although their adequacy is important to maintain a positive nitrogen (N) balance and prevent malnutrition. We carried out 42 studies to estimate N balance in 31 children over 3 years on APD for 19.8+/-15.7 months. Twenty metabolic studies were performed in patients dialysed for less than 1 year (7.2+/-3.3 months) and 22 in patients treated for more than 1 year (31.3+/-13.6 months). The mean estimated N balance of all metabolic studies was 57.5+/-62.8 mg/kg per day. In only 21 of 42 studies was N balance estimated to be over 50 mg/kg per day, which is considered adequate to meet N requirements for all metabolic needs and growth of uremic children. Estimated N balance correlated significantly with dietary protein intake (r=0.671, P=0.0001) and total energy intake (r=0.489, P=0.001). Using the equations of correlation, the values of dietary protein intake [=144% recommended dietary allowance (RDA)] and total energy intake (89% RDA) required to obtain an estimated N balance >50 mg/kg per day were calculated. Significantly lower estimated N balance values were obtained in the studies performed on patients on APD for over 1 year (36.09+/-54.02 mg/kg per day) than in patients treated for less than 1 year (81.11+/-64.70 mg/kg per day). In conclusion, based on the values of estimated N balance, we were able to establish adequate dietary protein and energy requirements for children on APD.

The contribution of nitric oxide to renal vascular wall thickening in rats with L-NAME-induced hypertension.

We investigated the mechanisms of renal vascular wall thickening in a rat model of N-nitro L-arginine methyl ester (L-NAME)-induced hypertension. To separate the effects of L-NAME-induced hypertension from other effects of nitric oxide (NO) inhibition, we created two models of L-NAME-induced hypertension: both had the same blood pressure level but NO inhibition was moderate in one group (group M) and severe in the other (group S). Urinary excretion of nitrates and nitrites was lower in group S than in group M. Wall thickening and lipid deposition in renal vessels were significantly greater in group S than in groups M. Simple and multiple regression analyses indicated that renal vascular wall thickening was more strongly correlated with lipid deposition than with blood pressure. The number of vessels positive for staining with Sudan black B was negatively correlated with urinary NO excretion. Expression of fibronectin and transforming growth factor-beta was greater in the Sudan black B-positive than in the Sudan black B-negative vessels, suggesting that extracellular matrix production was increased in vessels with lipid deposition. Lipid deposition and increased production of extracellular matrix may contribute to renal vascular wall thickening in L-NAME-induced hypertension. Some mechanisms independent of hypertension play important roles in vascular wall thickening induced by NO inhibition.

Analysis of nitroaromatic compounds in urine by gas chromatography-mass spectrometry for the biological monitoring of explosives.

Organic nitrocompounds are the most frequently used constituents of explosives and some of them have been evaluated to be highly toxic or even carcinogenic. Human contact with explosives may originate from a variety of sources, including occupational exposure during the production of ammunition as well as environmental exposure due to the contamination of soil and ground water reservoirs on former military production sites and training areas. This paper describes two gas chromatography-mass spectrometry-selected ion monitoring methods for the determination of twelve nitroaromatic compounds in urine (nitrobenzene, 1,2-dinitrobenzene, 1,3-dinitrobenzene, 1,3,5-trinitrobenzene, 2-nitrotoluene, 3-nitrotoluene, 4-nitrotoluene, 2,4-dinitrotoluene, 2,6-dinitrotoluene, 2,4,6-trinitrotoluene, 2-amino-4,6-dinitrotoluene, 4-amino-2,6-dinitrotoluene). The analytes are detectable in the lowest microg/l range, with imprecisions of 3-22% within series and 5-29% between series, depending on the compound of interest. Both procedures are rapid and relatively easy to perform and, therefore, are advantageous for the screening of occupationally or environmentally exposed persons. We analysed urine samples obtained from nine workers from an ammunition dismantling workshop and from twelve control persons. 2,4,6-Trinitrotoluene was detected in six samples at concentrations between 4 and 43 microg/l. The main metabolites of 2,4,6-trinitrotoluene, 4-amino-2,6-dinitrotoluene and 2-amino-4,6-dinitrotoluene, were found in a concentration range from 143 to 16,832 microg/l and from 24 to 5787 microg/l, respectively. Nonconjugated aminodinitrotoluenes were present as varying percentages of the total amount. 2,4-Dinitrotoluene and 2,6-dinitrotoluene were found in two samples (2-9 microg/l). Nitroaromatics were not detectable in urine specimens from control persons.

Chronic metabolic acidosis decreases albumin synthesis and induces negative nitrogen balance in humans.

Chronic metabolic acidosis has been previously shown to stimulate protein degradation. To evaluate the effects of chronic metabolic acidosis on nitrogen balance and protein synthesis we measured albumin synthesis rates and urinary nitrogen excretion in eight male subjects on a constant metabolic diet before and during two different degrees of chronic metabolic acidosis (NH4Cl 2.1 mmol/kg body weight, low dose group, and 4.2 mmol/kg body weight, high dose group, orally for 7 d). Albumin synthesis rates were measured by intravenous injection of [2H5ring]phenylalanine (43 mg/kg body weight, 7.5 atom percent and 15 atom percent, respectively) after an overnight fast. In the low dose group, fractional synthesis rates of albumin decreased from 9.9 +/- 1.0% per day in the control period to 8.4 +/- 0.7 (n.s.) in the acidosis period, and from 8.3 +/- 1.3% per day to 6.3 +/- 1.1 (P < 0.001) in the high dose group. Urinary nitrogen excretion increased significantly in the acidosis period (sigma delta 634 mmol in the low dose group, 2,554 mmol in the high dose group). Plasma concentrations of insulin-like growth factor-I, free thyroxine and tri-iodothyronine were significantly lower during acidosis. In conclusion, chronic metabolic acidosis causes negative nitrogen balance and decreases albumin synthesis in humans. The effect on albumin synthesis may be mediated, at least in part, by a suppression of insulin-like growth factor-I, free thyroxine and tri-iodothyronine.

Partition of nitrogenous substances in the urine of sheep on different dietary protein intakes.

The distribution of nitrogenous substances in urine was studied in sheep fed high (28.71 g N daily) and low (9.32 g N daily) protein diets for 6-8 weeks. The concentrations of total nitrogen and urea, allantoin, free amino acids, uric acid, hippuric acid, creatinine, creatine and ammonia were measured in urine. The nitrogen content of these measured substances was calculated. The results showed that the concentrations of total urinary nitrogen and nitrogen of individual nitrogenous substances were lower in sheep on a low protein diet, except for uric acid nitrogen which was unchanged. Urea nitrogen was responsible for up to 74% of the total urinary nitrogen of both groups of animals without any significant difference. The individual nitrogen portions of nitrogenous substances other than urea in total urinary-N did not exceed 7% during the intake of both diets. The intake of a low protein diet resulted in a significant increase in the proportion of nitrogen from uric acid, hippuric acid, creatinine and free amino acids, while the fraction of ammonia nitrogen in the total urinary N was significantly reduced. It is concluded that different protein or nitrogen intake in the diet do not have any significant effect on the urea nitrogen fraction in the total urinary nitrogen of sheep.