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Determination of phosphorus in small amounts of protein samples by ICP-MS.

Inductively coupled plasma mass spectrometry (ICP-MS) is used for phosphorus determination in protein samples. A small amount of solid protein sample (down to 1 micro g) or digest (1-10 micro L) protein solution was denatured in nitric acid and hydrogen peroxide by closed-microvessel microwave digestion. Phosphorus determination was performed with an optimized analytical method using a double-focusing sector field inductively coupled plasma mass spectrometer (ICP-SFMS) and quadrupole-based ICP-MS (ICP-QMS). For quality control of phosphorus determination a certified reference material (CRM), single cell proteins (BCR 273) with a high phosphorus content of 26.8+/-0.4 mg g(-1), was analyzed. For studies on phosphorus determination in proteins while reducing the sample amount as low as possible the homogeneity of CRM BCR 273 was investigated. Relative standard deviation and measurement accuracy in ICP-QMS was within 2%, 3.5%, 11% and 12% when using CRM BCR 273 sample weights of 40 mg, 5 mg, 1 mg and 0.3 mg, respectively. The lowest possible sample weight for an accurate phosphorus analysis in protein samples by ICP-MS is discussed. The analytical method developed was applied for the analysis of homogeneous protein samples in very low amounts [1-100 micro g of solid protein sample, e.g. beta-casein or down to 1 micro L of protein or digest in solution (e.g., tau protein)]. A further reduction of the diluted protein solution volume was achieved by the application of flow injection in ICP-SFMS, which is discussed with reference to real protein digests after protein separation using 2D gel electrophoresis.The detection limits for phosphorus in biological samples were determined by ICP-SFMS down to the ng g(-1) level. The present work discusses the figure of merit for the determination of phosphorus in a small amount of protein sample with ICP-SFMS in comparison to ICP-QMS.

Animals↗

Adverse effects of liquid protein fast on the handling of magnesium, calcium and phosphorus.

We have studied the effect of a vitamin- and potassium-supplemented liquid protein fast on mineral metabolism of six obese subjects (five women, 1 man) for 40 days. Each patient was admitted to a metabolic ward and was given daily 300 Kcal, 75 mg of calcium, 406 mg of phosphorus, 7 mg of magnesium, 33 meq of potassium, and 11.5 g of nitrogen. Urinary calcium, phosphorus and magnesium levels were greatest during the first week, but decreased as the fast continued to 21, 31 and 300 percent, respectively, above intake. Cumulative urinary losses of calcium, phosphorus and magnesium were 58, 75 and 500 percent greater, respectively, than the cumulative intake. Fecal losses for calcium, phosphorus and magnesium were less than urinary losses throughout the study. Cumulative fecal losses of magnesium were more than 30 percent greater than dietary intake. Mean daily balances were -104 mg (calcium), -48 mg (magnesium) and -363 mg (phosphorus). Serum phosphorus and magnesium levels did not change. However, serum calcium levels decreased (-0.5 mg/dl, p less than 0.05). Serum bicarbonate levels decreased 20 percent during the first 8 days of the fast, at which time urinary ammonium was maximal, but later returned to control values despite sustained increases in serum and urinary acids throughout the fast. Ammonium excretion was 260 to 300 percent above control values. Urinary titratable acid excretion was greatest early in the fast but subsequently decreased as the excretion of phosphorus declined. Titratable acid accounted for less of the excreted acid (7 to 21 percent) than did ammonia (70 to 90 percent). It is concluded that a liquid protein fast results in negative mineral balance that is not reflected by serum values and is due primarily to renal losses. The losses of magnesium were proportionally greater than those of calcium and phosphorus. These studies indicate that a liquid protein fast results in depletion of the intracellular and/or skeletal stores of these minerals.

Adult↗

Uneven hepatic iron and phosphorus distribution in beta-thalassemia.

BACKGROUND/AIMS: Determination of hepatic iron concentration is crucial in the evaluation of iron-storage disease. Iron content is normally determined in a part of a needle liver biopsy and the value obtained is considered to be representative of the iron concentration in the whole liver. To evaluate the reliability of this procedure, we studied iron distribution in the liver of two beta-thalassemic patients. Since the transport of intracellular iron is mediated by phosphates, we also studied the hepatic phosphorus distribution. METHODS: At autopsy, a liver slice extending from the left to the right lobe was divided into 51 and 49 samples, respectively. Each specimen was subdivided into two parts: one of them was paraffin-embedded and utilized for the histochemical detection of iron; the second part was analyzed for iron and phosphorus content by induced coupled plasma atomic emission spectroscopy. RESULTS: The histological picture of both livers was characterized by portal and periportal fibrosis associated with iron storage of different degree, without cirrhosis. The mean iron concentration of the liver was 20,631 +/- 4903 micrograms per g of dry tissue (micrograms/g dt) and 13,901 +/- 1976 micrograms/g dt, respectively. A striking variability in iron content between samples was also found: iron concentration ranged from 11,537 to 32,347 micrograms/g dt in the first case and from 6257 to 16,493 in the second case. We even observed regional differences in iron concentration, with a preferential peripheral accumulation in both cases and a tendency of the left compartment of the liver to accumulate more iron in the first case. Histochemical analyses confirmed the uneven iron distribution even at the acinar level, showing iron mainly being stored in hepatocytes and Kupffer cells of zone 1 of the acinus, with decreasing amounts of iron in zones 2 and 3. The mean hepatic phosphorus concentration was 6662 +/- 1300 micrograms/g dt (range: 4348-9947) and 7502 +/- 986 micrograms/g dt (range: 5844-90,282), respectively. The regional distribution of phosphorus was similar to that observed for iron. A strict correlation between iron and phosphorus content was also observed. CONCLUSIONS: Our data show that: 1) iron and phosphorus are unevenly distributed in the beta-thalassemic liver, even in the non-cirrhotic stages; 2) a regional pattern of iron and phosphorus distribution is evident, characterized by higher concentrations at the periphery of the liver; 3) the observed uneven distribution of iron and phosphorus implies that their content determined in a small liver sample cannot be considered as absolutely representative of the mean hepatic iron concentration. Therefore, iron concentrations determined in a part of a needle liver biopsy should be interpreted with caution in monitoring the efficacy of the iron-chelating therapy in beta-thalassemic patients.

Adult↗

The effects of sodium fluoride, calcium phosphate, and vitamin D2 for one to two years on calcium and phosphorus metabolism in postmenopausal women with spinal crush fracture osteoporosis.

Calcium and phosphorus balances and 47Ca turnover studies were performed before and after 12-27 months of daily treatment with sodium fluoride (60 mg), calcium (30-45 mmol), phosphate (29-44 mmol), and vitamin D2 (18,000 IU) in 20 postmenopausal women with spinal crush fracture osteoporosis. Before treatment the mean calcium balance (-1.6 mmol Ca/day) was negative (P less than 0.05), whereas the mean phosphorus balance (-1.6 mmol P/day) did not differ from zero (P greater than 0.10). The treatment induced an increase in calcium absorption (P less than 0.01) with an unchanged renal excretion and dermal loss of calcium and an increase (P less than 0.02) in calcium balance, which became positive (P less than 0.01) (+3.3 mmol Ca/day). This enhancement in calcium balance was obtained by the combined effects of an increase (P less than 0.05) in bone mineralization rate and a decrease (P less than 0.05) in bone resorption rate. The observed increase in net phosphorus absorption (P less than 0.001) was more pronounced than the increase in renal excretion of phosphorus (P less than 0.001), and the phosphorus balance therefore increased (P less than 0.001) and became positive (P less than 0.001). A positive correlation was observed between net calcium and net phosphorus absorption (r = 0.065; (P less than 0.001) in the 32 studies where both calcium and phosphorus balance data were available. Furthermore, a positive correlation (r = 0.61; P less than 0.05) was found between the changes in net calcium and net phosphorus absorption during treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Distribution and partitioning of phosphorus in solid waste and sediments from drainage canals in the industrial belt of Delhi, India.

Phosphorus is an essential and often limiting nutrient in both marine and freshwater ecosystems. However, its oversupply is of great concern in many environments due to its role in eutrophication. The concentration and distribution of sediment bound phosphorus species not only record its input into the environment, but also suggest their possible sources. It is recognized that the features of phosphorus behaviour have not been extensively studied in urban environments. In this study, phosphorus in solid waste and sediments from drainage canals within the industrial belt of Delhi were fractionated into five operationally defined forms, i.e., exchangeable or loosely sorbed phosphorus (Exch-P), Fe-bound phosphorus (Fe-P), authigenic apatite, CaCO3-bound phosphorus and biogenic apatite (Acet-P), detrital apatite (Det-P) and organic-bound phosphorus (Org-P), in order to assess the potential bioavailability status in these systems.

Apatites↗

Treatment of hypophosphatemia using a protocol based on patient weight and serum phosphorus level in a surgical intensive care unit.

BACKGROUND: Hypophosphatemia may cause organ derangements in the surgical intensive care unit. The purpose of this study was to determine the impact of a repletion protocol for hypophosphatemia based on admission weight and phosphorus level. STUDY DESIGN: All patients who presented to an 18-bed surgical intensive care unit with a serum phosphorus level of 2.2 mg/dL or less or who received phosphorus supplementation despite having normal levels were identified. In the preintervention phase between January and June 2001, 137 patients were retrospectively identified who met these criteria. A protocol was then designed giving a single intravenous dose of phosphorus based on weight and serum phosphorus. Repletion was given with sodium or potassium phosphorus based on presupplementation levels. After protocol implementation 141 patients met these criteria between September 2001 and February 2002, and treatment and postrepletion levels were followed prospectively. RESULTS: A total of 47 patients were repleted before the intervention with adequate followup and 22 (47%) attained a normal level. Supplementation success was 53% in moderate hypophosphatemia (2.2 mg/dL or less) and 27% in severe hypophosphatemia (less than 1.5 mg/dL). After protocol implementation, 111 patients were repleted with 84 (76%) correcting to a normal level (p = 0.002 compared with retrospective patients). Success was 78% in moderate hypophosphatemia and 62% in severe hypophosphatemia. Inappropriate supplementation of normal phosphorus levels decreased from 51 to 16 patients after protocol implementation. CONCLUSIONS: A protocol based on weight and serum levels successfully treated both moderate and severe hypophosphatemia in the majority of critically ill patients. Protocol implementation also decreased unnecessary supplementation of normal phosphorus levels.

Adult↗

Characterization of phosphorus-releasing bacteria in a small eutrophic shallow lake, Eastern China.

Phosphorus contents and phosphorus-releasing bacteria were characterized in a small eutrophic lake, Eastern China. Total phosphorus and water soluble inorganic phosphate (WSIP) in water bodies were as high as the levels of hyper-eutrophic lakes. Calcium bound phosphate and organic phosphorus were 2 major forms of phosphorus existing in the sediments. Enumeration of inorganic phosphate-solubilizing bacteria (IPB) and organic phosphorus-mineralizing bacteria (OPB) with culture-dependent method showed that these bacterial groups were not very rich in the ecosystem. Molecular identification and phylogenetic analysis of the predominant IPB and OPB strains indicated that there existed various kinds of bacteria participating in the phosphorus release. Laboratory tests on phosphorus release abilities showed IPB strains could liberate more than 50 microgml(-1), while OPB strains produced less than 2 microgml(-1) WSIP every day.

Bacteria↗

Adaptation to changes in dietary phosphorus intake in health and in renal failure.

Phosphate (Pi) homeostasis is maintained by the ability of the kidneys to adjust the tubular reabsorption of Pi to changes in the dietary intake of phosphorus. Renal tubular Pi reabsorption increases with the ingestion of a low-phosphorus diet (LPD) and decreases when a high-phosphorus diet (HPD) is consumed. A similar adaptive mechanism is also operative at the intestinal microvillus. The adaptive changes in Pi reabsorption are independent of parathyroid hormone production and are paralleled by similar changes in the Na+-dependent Pi transport at the brush border membrane (BBM). Type II Na+-Pi cotransporters (NaPi-2) are mainly involved in such regulatory mechanisms. Chronic dietary phosphorus restriction leads to increased Na+-Pi cotransport rate, along with increased NaPi-2 protein and mRNA abundance. In acute dietary phosphorus restriction, transport rate and NaPi-2 protein are also increased, but mRNA abundance remains unchanged. A shuttling mechanism involving translocation of cotransporters from intracellular pools to the BBM is involved in the rapid proximal tubular adaptation. The intestinal adaptation to changes in dietary phosphorus are similar to those described for the renal Pi transport, but the molecular structure of the intestinal Na+-Pi cotransporter is not known. When nephron mass is reduced, phosphate homeostasis is maintained through enhanced Pi excretion by residual nephrons. The adaptation to renal mass reduction is mediated by increased parathyroid hormone (PTH) production and by PTH-independent mechanisms, including increased intrarenal dopamine production. The adaptive changes of Pi transport to dietary phosphorus restriction can counteract the effect of dietary phosphorus reduction often prescribed in patients with renal failure. However, because of the reduced filtered load of Pi, the overall impact on serum Pi concentration is minimal.

Adaptation, Physiological↗

Effect of parenteral calcium and phosphorus therapy on mineral retention and bone mineral content in very low birth weight infants.

HYPOTHESIS: If calcium and phosphorus are administered to very low birth weight infants in amounts larger than those currently used in standard parenteral nutrition solutions, apparent retention of calcium and phosphorus (intake minus urinary excretion) will increase and bone mineralization will improve. DESIGN: Randomized, controlled, double-blind trial. SETTING: Neonatal intensive care unit. PATIENTS: Twenty-four very low birth weight infants (< 1.2 kg) expected to receive parenteral nutrition exclusively for approximately 3 weeks beginning 3 days after birth. INTERVENTIONS: Infants received parenteral nutrition solutions, either the standard mixture containing 1.25 mmol calcium and 1.5 mmol phosphorus per deciliter (group STAND: n = 12, birth weight 921 +/- 171 gm, gestational age 27 +/- 2 weeks (mean +/- SD)) or 1.7 mmol calcium and 2.0 mmol phosphorus per deciliter (group HIGH: n = 12, 857 +/- 180 gm, 27 +/- 2 weeks). MAIN OUTCOME MEASURES: Intake, urinary excretion, and apparent retention of calcium, phosphorus, and magnesium every 3 days during parenteral nutrition therapy. Serum indexes of mineral status twice during therapy. Bone mineral content of the distal segment of the left radius at 1, 4, 8, and 26 weeks. RESULTS: Apparent calcium retention (1.2 +/- 0.2 vs 1.6 +/- 0.2 mmol.kg-1.d-1) and phosphorus retention (1.4 +/- 0.2 vs 1.8 +/- 0.4 mmol.kg-1.d-1) differed significantly (p < 0.01) between groups STAND and HIGH, respectively; neither changed with the duration of parenteral nutrition therapy. Serum calcium, magnesium, parathyroid hormone, 25-hydroxyvitamin D, and osteocalcin concentrations were similar in both groups. Serum phosphorus concentration was significantly higher in group HIGH than in group STAND (p = 0.025). The absolute bone mineral content and the rate of increase in bone mineral content between 1 and 4, 1 and 8, and 1 and 26 weeks were significantly greater in group HIGH than in group STAND. CONCLUSIONS: Increased parenteral intakes of calcium and phosphorus resulted in greater retention of these minerals during parenteral nutrition therapy and in greater bone mineral content after therapy.

Bone Density↗

Metabolism in vitro of 25-hydroxycholecalciferol in chicks fed on phosphorus-deficient diets.

1. Three groups of 10-days-old chicks were fed on one of three diets having phosphorus contents of 0.08 mol/kg, 0.14 mol/kg or 0.21 mol/kg. Ten days later duodenal calcium absorption by the ligated loop technique in vivo, and plasma calcium and phosphorus concentrations, were measured. In addition the metabolism in vitro of 25-hydroxycholecalciferol [25-(OH)D3] by kidney homogenates was studied. 2. In the low phosphorus group (0.08 mol/kg) calcium absorption and the activity of 25-(OH)D3-1-hydroxylase were significantly higher than those of the high phosphorus group (0.21 mol/kg). However, in the medium phosphorus group (0.14 mol/kg), calcium absorption was significantly higher although the activity of 25-(OH)D3-1-hydroxylase was not significantly higher when compared with the high phosphorus group (0.21 mol/kg). 3. It is concluded that in phosphorus deprivation, unlike in calcium deprivation, a diet very low in phosphorus is required to stimulate the renal 25-(OH)D3-1-hydroxylase activity.

25-Hydroxyvitamin D3 1-alpha-Hydroxylase↗

Psr1, a nuclear localized protein that regulates phosphorus metabolism in Chlamydomonas.

Understanding the ways in which phosphorus metabolism is regulated in photosynthetic eukaryotes is critical for optimizing crop productivity and managing aquatic ecosystems in which phosphorus can be a major source of pollution. Here we describe a gene encoding a regulator of phosphorus metabolism, designated Psr1 (phosphorus starvation response), from a photosynthetic eukaryote. The Psr1 protein is critical for acclimation of the unicellular green alga Chlamydomonas reinhardtii to phosphorus starvation. The N-terminal half of Psr1 contains a region similar to myb DNA-binding domains and the C-terminal half possesses glutamine-rich sequences characteristic of transcriptional activators. The level of Psr1 increases at least 10-fold upon phosphate starvation, and immunocytochemical studies demonstrate that this protein is nuclear-localized under both nutrient-replete and phosphorus-starvation conditions. Finally, Psr1 and angiosperm proteins have domains that are similar, suggesting a possible role for Psr1 homologs in the control of phosphorus metabolism in vascular plants. With the identification of regulators such as Psr1 it may become possible to engineer photosynthetic organisms for more efficient utilization of phosphorus and to establish better practices for the management of agricultural lands and natural ecosystems.

Adaptation, Physiological↗

Evaluation of the dietary interaction of calcium and phosphorus in the high producing laying hen.

1. In a 6 x 7 factorial experiment using 2688 22-week-old laying hens of the Lohmann-SL strain kept in cages (4 birds/cage), diets containing six calcium (20, 25, 30, 35, 40, 45 g calcium/kg) and seven phosphorus concentrations (3.2, 4.2, 5.2, 6.2, 7.2, 8.2, 16.2 g total phosphorus/kg (Pt)) were combined orthogonally. The resulting 42 treatments were replicated 8 times so that a replicate consisted of a double cage of 2 x 4 hens. The experiment lasted 40 weeks (10 x 28 days). 2. The experimental diets, based on maize and soyabean meals contained 11.5 MJ metabolisable energy/kg and 175 g/kg protein. Different dietary calcium and phosphorus contents were obtained by substituting oat hulls with limestone and dicalcium phosphate. 3. Mortality, egg production, egg weight, egg mass, food intake and food conversion efficiency were determined as well as the breaking strength, thickness of shells and the percentage of eggs with defective shells. 4. All responses measured were significantly influenced by the variance sources (calcium, phosphorus, interaction). Most of the production traits responded asymptotically to increasing dietary phosphorus concentration, the greatest increases or decreases generally being seen between 3.2 and 5.2 g Pt/kg. Further but weaker increases were seen between 5.2 and 8.2 or 16.2 g Pt/kg. 5. Increases in dietary calcium content always resulted in curvilinear responses. In all cases optimal effects were obtained with diets containing 25 g calcium/kg and the worst values at 45 g calcium/kg. The interaction between calcium and phosphorus was recognised by strong performance depressions and a high mortality at combinations of the lowest phosphorus concentration (3.2 g/kg) with high calcium contents (35 to 45 g/kg). These were largely offset by increasing dietary phosphorus. Thus, between 7.2 and 16.2 g Pt/kg and 25 and 45 g Ca/kg a plateau was formed where only small differences in egg production were observed. 6. From the three egg shell characteristics measured, breaking strength and shell thickness responded differently to the percentage of eggs with defective shells. While breaking strength and shell thickness were respectively negatively and positively influenced by increasing dietary phosphorus and calcium contents, both elements affected the proportion of eggs with defective shells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Evaluation of Activated Sludge Model No. 2 at high phosphorus concentrations.

This paper presents laboratory scale experimentation carried out to study enhanced biological phosphorus removal at high phosphorus concentrations in a sequencing batch reactor. Four series of data obtained in a sequencing batch reactor are examined in light of the Activated Sludge Model No. 2. This model was calibrated using data from the first and second series working at low phosphorus concentrations. The Activated Sludge Model No. 2 successfully characterised the enhanced biological phosphorus removal performance of the sequencing batch reactor at low phosphorus concentrations. The calibrated model was then used to adjust experimental results of the other series working at high phosphorus concentration. Differences between model predictions and experimental data could be explained by redissolution in the anaerobic phase and precipitation in the aerobic phase of calcium phosphates not taken into account by the model. This hypothesis can be justified by the conditions of pH and phosphorus concentration prevailing in the experiments and it was confirmed by the results of another experiment in which precipitated phosphorus profile was measured during the cycle.

Bioreactors↗

Phosphorus defficiency and sludge bulking.

Separation by settling is one of the most troublesome stages of an activated sludge process. A decrease in the efficiency of separation of microbial biomass from the treated effluent causes a decrease in the overall efficiency of the treatment plant. One of the most common problems in activated sludge systems to negatively influence the settleability is sludge bulking which can be defined as non-settling situation of microbial mass. The main objective of this study is to find out the effect of phosphorus deficiency on bulking of activated sludge. In order to achieve this aim, the effects of a strictly phosphorus limited environment was investigated by running 6 semi-continuous activated sludge reactors. Since the specific aim of the study is to identify the effects of magnesium and calcium ions on sludge bulking in a phosphorus deficient medium, 3 different magnesium and calcium concentrations (5, 10, 20 meq l(-1)) were studied. Laboratory scale reactors were fed with synthetic wastewater and operated at a sludge age of 8 days. Results showed that, phosphorus deficiency caused sludge bulking. Sludge samples examined under microscope showed that the floc structures from reactors fed with different cations are significantly different. To identify the differences, a series of chemical analyses were conducted on the sludge samples. In addition to that, sludge volume index (SVI) values were monitored on a daily basis. Next, the concentration of phosphorus in the feed medium was increased in two separate steps and the possibility to cure the sludge bulking problem was investigated. It was found that bulking of activated sludges due to phosphorus deficiency could be cured by the addition of phosphorus. For phosphorus deficient bulking in relation to the cations present in the system some mechanisms are suggested.

Biomass↗

Study on the recovery of phosphorus from waste-activated sludge incinerator ash.

Several batch studies that were made up of the acid extraction and the solvent extraction were performed to recover phosphorus from the waste-activated sludge (WAS) incinerator ash. In the acid extraction, the extraction efficiency of phosphorus relied on the acid type, liquid(acid)-to-solid (L(acid)S) ratio, and acid concentration. Phosphorus in the WAS incinerator ash was completely extracted by 1 M HCl at the L(acid)S ratio of 6.4:1. Subsequently, the solvent extraction was conducted to separate and concentrate phosphorus further from the acid extract. The efficiency of solvent extraction was affected mainly by the solvent type, liquid (solvent)-to-liquid (the acid extract) (L(solv)L(acid ext)) ratio, and hydrogen ion concentration. Under the appropriate condition, 76% of phosphorus in the acid extract was extracted to 1-butanol phase, which corresponded to 80.1% as the mass fraction of phosphorus to total elements. Prior to the solvent extraction, the addition of bis (2-ethylhexyl) phosphoric acid (D2EHPA), which was available for removing aluminum from the acid extract, led to an additional increase in the term of the mass fraction of phosphorus to total elements. Overall results indicated that phosphorus in the WAS incinerator ash could be efficiently recovered and be a potential renewable resource.

Hydrochloric Acid↗

Age-related changes of plasma alkaline phosphatase and inorganic phosphorus, and late ossification of the cranial roof in the Spanish imperial eagle (Aquila adalberti C. L. Brehm, 1861).

Plasma alkaline phosphatase and inorganic phosphorus levels were determined for 52 nestling Spanish imperial eagles from two wild populations and 22 captive adults and subadults (10 adults and 12 subadults). The exact age was known for all birds. Mean alkaline phosphatase and inorganic phosphorus were higher in chicks than in the captive adults and subadults. Sex differences were not observed, and nestlings from different populations showed similar values. No significant regression described the relationship between age and alkaline phosphatase or inorganic phosphorus throughout the nestling period. However, alkaline phosphatase and inorganic phosphorus decreased significantly throughout the subadult period, with age explaining 98.2% and 50.5% of the variation in alkaline phosphatase and inorganic phosphorus levels, respectively. Non-fully-ossified zones were measured in frontal bones of another 12 subadult eagles that died at known ages. Ossification increased throughout the subadult period and was significantly correlated with expected levels of alkaline phosphatase or inorganic phosphorus (i.e., values predicted from the regression equations derived from the first analysis). Minimum alkaline phosphatase levels and full ossification of the cranial roof coincided with puberty onset. We conclude that, in subadult Spanish imperial eagles, decreasing alkaline phosphatase and inorganic phosphorus values are related to the ossification of frontal bones, although a contribution of other unknown processes of late ossification cannot be excluded, and alkaline phosphatase (but not inorganic phosphorus) may be a useful parameter for age-predicting purposes.

Aging↗

Effect of dietary protein and phosphorus levels on the utilization of zinc, copper and manganese by adult males.

Zinc, copper and manganese utilization were examined in eight adult males fed a low protein-moderate phosphorus diet (LPrMP), a low protein-high phosphorus diet (LPrHP), a high protein-moderate phosphorus diet (HPrMP) and a high protein-high phosphorus diet (HPrHP) during a 51-day balance study. The low and high protein diets contained 8.1 g and 24.1 g nitrogen daily, respectively. The moderate and high levels of phosphorus were 1,010 and 2,525 mg phosphorus daily. Subjects lost less zinc in the feces when fed the HPrMP diet than when fed the other three diets. The level of dietary protein and phosphorus all statistically affected fecal zinc excretion. Urinary zinc excretion was significantly greater when subjects consumed the high protein diets rather than the low protein diets. Apparent retention of zinc was greatest when subjects were fed the HPrMP diet rather than the other three diets. This effect was statistically attributable to the interaction between dietary protein and phosphorus. Serum zinc levels were significantly elevated when subjects consumed the high protein rather than the low protein diets. Serum zinc levels were correlated to urinary zinc excretion (r = 0.788, P< 0.001), apparent retention of zinc (r = 0.385, P< 0.05) and serum copper levels (r = 0.395, P < 0.05). Apparent absorption and retention of copper were significantly greater when subjects were fed the high protein rather than the low protein diets. They dietary treatments did not affect the urinary excretion of copper, serum copper levels or the apparent absorption and retention of manganese by these subjects.

Adult↗

Nutritional interrelationships between calcium, phosphorus and lactose in rats.

From the age of 3 months, six groups of rats consisting of six animals each were fed one of three types of diet with either 15% lactose or dextrose. The diets used were a control diet containing 0.6% calcium and 0.45% phosphorus and two high-phosphorus (1.3%) diets with either a normal (0.6%) or a low (0.2%) calcium content. Whereas calcium retention and femur mass appeared to be unaffected, feeding the high-phosphorus diets resulted in significant (P less than or equal to 0.05 or P less than or equal to 0.01) biochemical changes including: decreased plasma phosphorus almost throughout the whole of the study; increased plasma alkaline phosphatase, and urinary total hydroxyproline after 17 and 42 weeks; nephrocalcinosis; depressed urinary calcium, and reduced femur density. With the excretion of nephrocalcinosis and depressed urinary calcium, these changes were more marked if the calcium content of the diet was low than when it was normal, and this difference was associated with a significant (P less than 0.01) inhibiting effect of dietary calcium on the intestinal absorption and urinary excretion of phosphorus. Although lactose, as compared with dextrose, significantly (P less than 0.01) improved the intestinal absorption and retention of calcium during body weight gain, which was reflected by an increased femur mass after 42 weeks, this sugar did not reduce the detrimental effects on bone and soft tissue resulting from feeding the high-phosphorus diets. The results of the study in line with the induction of nutritional secondary hyperparathyroidism and increased bone turnover in rats fed high-phosphorus diets and indicate that lactose-stimulated calcium absorption will not prevent or diminish the biochemical changes associated with this disease; the results also stress the significance of the calcium content of the diet as a factor that may protect the body against excessive dietary phosphorus.

Alkaline Phosphatase↗