Determination of vitamins D,A, and E in sera and vitamin D in milk from captive and free-ranging polar bears (Ursus maritimus), and 7-dehydrocholesterol levels in skin from captive polar bears

The difference between serum levels from 36 captive and 56 free-ranging polar bears (Ursus maritimus) for 25-hydroxyvitamin D (25-OH-D) was found not to be significant (mean ± SD = 348 ± 215 nmol/L [captive], 360 ± 135 nmol/L [free-ranging], t = 0.30, df = 52.8, P = 0.76), whereas the difference for retinol and α-tocopherol was significant (retinol, 1.37 ± 0.67 μmol/L [captive] 1.89 ± 0.63 μmol/L [free-ranging], t = 3.88, df = 72.4, P <0.001, α-tocopherol, 18.56 ± 18.56 μmol/L [captive], 48.76 ± 13.92 μmol/L [free-ranging], t = 7.85, df = 61.9, P < 0.001). Due to the high fat content in the polar bear diet, seal blubber may be the source of these fat-soluble vitamins. Six skin biopsies were analyzed from captive polar bears at the Denver Zoological Gardens for 7-dehydrocholesterol levels and found to contain 0.11 ± 0.03 nmol/cm². This finding also helps to support the contention that the source of vitamin D for polar bears may be ingestion and not cutaneous production. Vitamin D content in the milk from one captive sow in the den (0.14 nmol/g) and 10 free-ranging sows with cubs of the year out on the ice pack (0.0042 ± 0.0073 nmol/g) were also evaluated. It would be helpful to evaluate additional milk samples from denning and non-denning sows with cubs to see whether vitamin D content varies according to the stage of lactation. Zoo Biol 17:285–293, 1998. © 1998 Wiley- Liss, Inc.     

Milk production and composition in reindeer (Rangifer tarandus): effect of lactational stage

Milk yield and composition of major milk constituents were measured in captive, nursing reindeer. Registration of milk production was performed during two successive lactations (2001 and 2002). The milk yield was significantly affected by week of lactation (P<0.001) and by individual (P<0.001). The lactation curve had an asymmetrical peak 3 weeks postpartum and the milk yield at peak lactation was 983 g/day (range 595-1239). The length of lactation varied from 24 to 26 weeks and average total milk production was 99.5 kg. From peak lactation the milk production decreased linearly (P<0.001) until milk production was terminated. Mean values for content of major milk constituents were 15.5% fat, 9.9% protein and 2.5% lactose. The content of fat and protein increased markedly with the lactation stage (P<0.001), while lactose showed a slight decrease (P<0.001). The milk composition was significantly affected by stage of lactation (P<0.001). There was a marginally significant decrease in protein:fat ratio (P=0.06) as protein was substituted by fat with stage of lactation. The caloric value of the milk averaged 8.7 kJ/g and increased significantly with the stage of lactation (P<0.001). The overall increase in milk gross energy content during lactation was 67.6%. The energy output averaged 7996 kJ/day at peak lactation and decreased significantly during the course of lactation (P=0.002).     

Milk composition in free-ranging polar bears (Ursus maritimus) as a model for captive rearing milk formula

The goals of this study were to have an improved understanding of milk composition and to help create a suitable milk formula for cubs raised in captivity. Milk samples were evaluated for fat, fatty acids, carbohydrate, vitamin D(3), 25(OH)D(3), vitamin A (retinol), vitamin E (α-tocopherol), protein, and amino acids. Total lipids in milk did not differ for cubs (mean ± SEM = 26.60 ± 1.88 g/100 ml vs. yearlings 27.80 ± 2.20 g/100 ml). Milk lipids were of 23.6% saturated fatty acid for cubs and 22.4% for yearlings. Milk consumed by cubs and yearlings contained 43.8 and 42.0% mono-unsaturated fatty acids and 23.4 and 21.9% polyunsaturated fatty acids, respectively. Carbohydrate content was higher in milk for cubs (4.60 ± 0.64 g/100 ml) than for yearlings (2.60 ± 0.40 g/100 ml). Vitamin D(3) concentration of milk was 18.40 ± 5.00 ng/ml in early lactation compared with 7.60 ± 2.00 ng/ml for mid-lactation. 25(OH)D(3) was lower in milk consumed by cubs (162.00 ± 6.70 pg/ml) than in milk consumed by yearlings (205.00 ± 45.70 pg/ml). Vitamin A concentrations were 0.06 ± 0.01 and 0.03 ± 0.01 μg/ml for cubs and yearlings, respectively. Vitamin E was higher in milk consumed by cubs (20.16 ± 4.46 μg/ml) than by yearlings (7.30 ± 1.50 μg/ml). Protein content did not differ in milk available to cubs (11.40 ± 0.80 g/100 ml compared with milk for yearlings 11.80 ± 0.40 g/100 ml). Taurine was the most abundant free amino acid at 3,165.90 ± 192.90 nmol/ml (0.04% as fed basis).     

Alpha melanocyte stimulating hormone inhibits prolactin secretion in fetal and infant lambs

The intravenous administration of αMSH (25 μg/kg) to 11 lambs (3 to 29 days of age) suppressed plasma PRL by 15 minutes. The mean basal concentration was 15.3 ± 2.9 ng/ml and the mean nadir was 4.9 ± 0.8 ng/ml (p<0.01). In chronically catheterized fetuses (128–140 days), intravenous administration of αMSH (25 μg/kg) decreased basal PRL levels (89.6 ± 12.4 ng/ml) significantly at 15–30 minutes to levels of 74.3 ± 11.4 ng/ml (p<.01). The degree of suppression of basal PRL levels was less in fetusus (76.9 ± 4.1%) than that induced in the neonates (40.5 ± 7.1%). In younger fetuses <120 days in whom basal PRL levels are low (3.0 ± 2.1 ng/ml), administration of αMSH was without effect. Plasma GH concentrations were not altered by administration of αMSH. The suppression of PRL secretion by αMSH administration could result from increased release of hypothalamic dopamine or be a direct effect on secretion of prolactin by the pituitary.     

High dietary vitamin A interferes with tissue α-tocopherol concentrations in fattening pigs: a study that examines administration and withdrawal times

This study aimed to assess the interaction between different dietary vitamin A (dVitA) levels and the same concentration of vitamin E (100 IU all-rac-α-tocopheryl acetate/kg feed) in growing-finishing pigs. In the first experiment, two fat sources × two dVitA levels (0 v. 100 000 IU) were used. The supplementation of 100 000 IU dVitA induced a range of 5.13 to 30.03 μg retinol/g liver, 62.78 to 426.88 μg retinol palmitate/g liver, and 0.60 to 1.96 μg retinol/g fat. Dietary fat did not affect retinol or retinyl palmitate deposition in pigs. The high concentration of dVitA produced lower fat and liver α-tocopherol concentrations, and increased susceptibility of muscle tissue to oxidation. A second experiment was carried out to study the retinol and α-tocopherol retention at different withdrawal times prior to slaughter (two dVitA levels; 0 v. 100 000 IU). A high dose of 100 000 IU vitamin A during a short 2-week period was enough to induce α-tocopherol depletion in liver and fat to a similar extent as when 100 000 IU were administered during the whole fattening. Muscle, fat and liver α-tocopherol concentrations were not affected by dVitA in the 1300-13 000 IU/kg range, but liver α-tocopherol concentration was higher when vitamin A was removed from the vitamin mix 5 weeks prior to slaughter (experiment 3).     

Plasma tocopherol,retinol, and carotenoid concentrations in free-ranging Humboldt penguins (Spheniscus humboldti) in Chile

Plasma retinol and α-tocopherol concentrations were measured in heparinized blood samples collected from 51 free-ranging adult Humboldt penguins (Sphenicus humboldti) residing at two colonies off the Chilean coast. Thirty samples were collected in April 1992 from penguins inhabiting the Ex-islote de los Pájaros Niños in Algarrobo, Chile. In September 1992, 21 samples were collected from birds inhabiting Isla de Cachagua, Chile. Samples were assayed for retinol, retinyl palmitate, α-tocopherol, γ-tocopherol, lutein, β-cryptoxanthin, lycopene, α-carotene, and β-carotene. Retinol, α-tocopherol, and lutein were detected in all samples, while lycopene and γ-tocopherol were not detected in any. A significantly higher percentage of samples had detectable levels of retinyl palmitate and α-carotene in April (P < 0.001): for β-cryptoxanthin the percentage was higher in September (P < 0.001). Plasma concentrations of α-tocopherol and lutein were higher in September. Alpha-tocopherol concentrations were 1,877.1 ± 99.0 (SEM) μg/dl in April compared to 2.289 ± 122.3 μg/dl in September (P < 0.05); lutein concentrations were 4.16 ± 0.43 μg/dl in April vs. 10.68 ± 1.02 μg/dl in September (P < 0.001). Retinol concentrations were not significantly different (117 ± 8.0 μg/dl in April vs. 105.3 ± 7.6 μg/dl in September). Both physiologic changes associated with season, and the change in locale may have contributed to the differences seen in the assay means and the number of samples with detectable levels. © 1996 Wiley-Liss, Inc.     

Effects of dietary energy intake during gestation and lactation on milk yield and composition of first,second and fourth parity sows

Abstract

In order to determine the effects of a varied level of dietary energy intake during pregnancy and lactation on milk yield and composition, first, second and fourth parity sows (Large White × German Landrace) were provided with energy at a level of either: (i) 100% of ME requirement (MEreq) during pregnancy and lactation, (ii) 120% MEreq during pregnancy and 80% during lactation, and (iii) 80% MEreq during pregnancy and 120% during lactation. In spite of equal target levels feed analysis revealed that gestating first parity sows with 120/80 treatment combination and lactating sows of 80/120 treatment combination received 25, and 11 – 17% more digestible N than in the respective 100/100 treatment combination. Irrespective of this 120/80 sows responded with the highest milk DM, fat, and energy contents, and the lowest lactose concentrations whereas protein levels where not affected, irrespective of parity (p < 0.05). Milk yield of sows in 1st and 4th lactation was 85 and 106% of that in 2nd lactation, respectively. Average milk composition was 18.1% DM, 4.9% protein, 6.8% fat, 5.6% lactose, and 0.8% ash. Milk composition changes ceased at day 7 of lactation with a reduction of milk GE and protein, and an increase of lactose content. Concentrations of threonine, arginine, valine, leucine, tyrosine, phenylalanine, cystine, and tryptophan, as well as stearic, oleic, and linoleic acid were higher in colostrum than in milk at later lactation stages. In contrast, laurine, myristic, palmitic, and palmitoleic acids were lower concentrated in colostrum. In conclusion, these results illustrate the importance of body reserve mobilization for milk production in sows and indicate that low energy supply during gestation cannot be compensated by higher energy supply during lactation.     

A comparison of milk composition in Myotis lucifugus and Eptesicus fuscus (chiroptera: vespertilionidae)

Milk composition in two species of insectivorous bats (Myotis lucifugus and Eptesicus fuscus) was determined. No significant differences in fat, lactose, protein, or energy content were evident during the lactation period for M. lucifugus. The percentage of fat, lactose, and protein averaged 13.5, 3.3, 7.4, respectively, for M. lucifugus, and 16.4, 2.5, 6.2, respectively, for E. fuscus. Energy content of milk averaged 7.32 kJ/g for M. lucifugus and 8.37 kJ/g for E. fuscus. The percentage of lactose in milk from M. lucifugus was significantly greater than in E. fuscus, but there were no significant interspecific differences in percentages of fat or protein. The two species also differed significantly in content of five out of nine fatty acids.     

Supplementing sow diets with palm oil during late gestation and lactation: effects on milk production,sow hormonal profiles and growth and development of her offspring

The supplementing of sow diets with lipids during pregnancy and lactation has been shown to reduce sow condition loss and improve piglet performance. The aim of this study was to determine the effects of supplemental palm oil (PO) on sow performance, plasma metabolites and hormones, milk profiles and pre-weaning piglet development. A commercial sow ration (C) or an experimental diet supplemented with 10% extra energy in the form of PO, were provided from day 90 of gestation until weaning (24 to 28 days postpartum) in two groups of eight multiparous sows. Gestation length of PO sows increased by 1 day (P<0.05). Maternal BW changes were similar throughout the trial, but loss of backfat during lactation was reduced in PO animals (C: −3.6±0.8 mm; PO: −0.1±0.8 mm; P<0.01). Milk fat was increased by PO supplementation (C day 3: 8.0±0.3% fat; PO day 3: 9.1±0.3% fat; C day 7: 7.8±0.5% fat; PO day 7: 9.9±0.5% fat; P<0.05) and hence milk energy yield of PO sows was also elevated (P<0.05). The proportion of saturated fatty acids was greater in colostrum from PO sows (C: 29.19±0.31 g/100 g of fat; PO: 30.77±0.36 g/100 g of fat; P<0.01). Blood samples taken on 105 days of gestation, within 24 h of farrowing, day 7 of lactation and at weaning (28±3 days post-farrowing) showed there were no differences in plasma concentrations of triacylglycerol, non-esterified fatty acids, insulin or IGF-1 throughout the trial. However, circulating plasma concentrations of both glucose and leptin were elevated during lactation in PO sows (P<0.05 and P<0.005, respectively) and thyroxine was greater at weaning in PO sows (P<0.05). Piglet weight and body composition were similar at birth, as were piglet growth rates throughout the pre-weaning period. A period of 7 days after birth, C piglets contained more body fat, as indicated by their lower fat-free mass per kg (C: 66.4±0.8 arbitrary units/kg; PO: 69.7±0.8 arbitrary unit/kg; P<0.01), but by day 14 of life this situation was reversed (C: 65.8±0.6 arbitrary units/kg; PO: 63.6±0.6 arbitrary units/kg; P<0.05). Following weaning, PO sows exhibited an increased ratio of male to female offspring at their subsequent farrowing (C: 1.0±0.3; PO: 2.2±0.2; P<0.05). We conclude that supplementation of sow diets with PO during late gestation and lactation appears to increase sow milk fat content and hence energy supply to piglets. Furthermore, elevated glucose concentrations in the sow during lactation may be suggestive of impaired glucose homoeostasis.     

RNA synthesis in imaginal disks of Galleria mellonella: Effects of alpha- and beta-ecdysone and fat body in vitro

The effects of alpha-ecdysone (α-E), beta-ecdysone (β-E), and larval fat body on morphogenesis and total RNA synthesis in wing imaginal disks of Galleria mellonella were studied. Both ecdysones induce morphogenesis of disks in vitro. Alpha-ecdysone and β-E (0·3–3·0 μg/ml) stimulate RNA synthesis 30 and 60 per cent above control levels, respectively. While less α-E (0·3 μg/ml) is required to increase RNA synthesis than to induce morphogenesis, the reverse is true for β-E. Morphogenesis (i.e. tracheole migration and evagination) can proceed in the presence of concentrations of β-E (0·03 μg/ml) that are subthreshold for the induction of RNA synthesis (0·3 μg/ml). We conclude, therefore, that the increase in total RNA (presumbly ribosomal) is unrelated to and not a prerequisite for tracheole migration or evagination. If morphogenically active preconditioned medium (i.e. medium in which α-E and fat body have been incubated for 48 hr and the fat body then removed) is added to disk cultures, RNA synthesis is not stimulated. Apparently, increases in total RNA caused by both ecdysones may not be necessary for early in vitro disk development. The independent nature of some ecdysone-induced events and implications of our conclusions are discussed.     

Genetic (co)variances between milk mineral concentration and chemical composition in lactating Holstein-Friesian dairy cows

Milk mineral concentration is important from both the perspective of processing milk into dairy products and its nutritive value for human consumption. Precise estimates of genetic parameters for milk mineral concentration are lacking because of the considerable resources required to collect vast phenotypes quantities. The milk concentration of calcium (Ca), potassium (K), magnesium (Mg), sodium (Na) and phosphorus (P) in the present study was quantified from mid-IR spectroscopy on 12 223 test-day records from 1717 Holstein-Friesian cows. (Co)variance components were estimated using random regressions to model both the additive genetic and within-lactation permanent environmental variances of each trait. The coefficient of genetic variation averaged across days-in-milk (DIM) was 6.93%, 3.46%, 6.55%, 5.20% and 6.68% for Ca, K, Mg, Na and P concentration, respectively; heritability estimates varied across lactation from 0.31±0.05 (5 DIM) to 0.67±0.04 (181 DIM) for Ca, from 0.18±0.03 (60 DIM) to 0.24±0.05 (305 DIM) for K, from 0.08±0.03 (15 DIM) to 0.37±0.03 (223 DIM) for Mg, from 0.16±0.03 (30 DIM) to 0.37±0.04 (305 DIM) for Na and from 0.21±0.04 (12 DIM) to 0.57±0.04 (211 DIM) for P. Genetic correlations within the same trait across different DIM were almost unity between adjacent DIM but weakened as the time interval between pairwise compared DIM lengthened; genetic correlations were weaker than 0.80 only when comparing both peripheries of the lactation. The analysis of the geometry of the additive genetic covariance matrix revealed that almost 90% of the additive genetic variation was accounted by the intercept term of the covariance functions for each trait. Milk protein concentration and mineral concentration were, in general, positively genetically correlated with each other across DIM, whereas milk fat concentration was positively genetically correlated throughout the entire lactation with Ca, K and Mg; the genetic correlation with fat concentration changed from negative to positive with Na and P at 243 DIM and 50 DIM, respectively. Genetic correlations between somatic cell score and Na ranged from 0.38±0.21 (5 DIM) to 0.79±0.18 (305 DIM). Exploitable genetic variation existed for all milk minerals, although many national breeding objectives are probably contributing to an indirect positive response to selection in milk mineral concentration.     

Elevated Insulin Sensitivity in Low‐protein Offspring Rats Is Prevented by a High‐fat Diet and Is Associated With Visceral Fat

This study tests the hypothesis that a high‐fat postnatal diet increases fat mass and reduces improved insulin sensitivity (IS) found in the low‐protein model of maternal undernutrition. Offspring from Wistar dams fed either a 20% (control (CON)) or 8% (low protein (LP)) protein diet during gestation and lactation were randomly assigned to a control (con) or cafeteria (caf) diet at weaning (21 days) until 3 months of age at which point IS was measured (hyperinsulinemic–euglycemic clamp). Fat mass, growth, energy intake (EI) and expenditure (EE), fuel utilization, insulin secretion, and leptin and adiponectin levels were measured to identify a possible role in any changes in IS. IS was increased in LP‐con in comparison to CON‐con animals. Cafeteria feeding prevented this increase in LP animals but had no effect in CON animals (insulin‐stimulated glucose infusion rates (GIRs; mg/min/kg); CON‐con: 13.9 ± 1.0, CON caf: 12.1 ± 2.1, LP‐con: 25.4 ± 2.0, LP‐caf: 13.7 ± 3.7, P < 0.05). CON‐caf animals had similar percent epididymal white adipose tissue (%EWAT; CON‐con: 1.71 ± 0.09 vs. CON‐caf: 1.66 ± 0.08) and adiponectin (µg/ml: CON‐con: 4.61 ± 0.34 vs. CON‐caf: 3.67 ± 0.18) except hyperinsulinemia and relative hyperleptinemia in comparison to CON‐con. Differently, LP‐caf animals had increased %EWAT (LP‐con: 1.11 ± 0.06 vs. LP‐caf: 1.44 ± 0.08, P < 0.05) and adiponectin (µg/ml: LP‐con: 5.38 ± 0.39 vs. LP‐caf: 3.75 ± 0.35, P < 0.05) but did not show cafeteria‐induced hyperinsulinemia or relative hyperleptinemia. An increased propensity to store visceral fat in LP animals may prevent the elevated IS in LP offspring.     

Mammary Specific Transgenic Over-expression of Insulin-like Growth Factor-I (IGF-I) Increases Pig Milk IGF-I and IGF Binding Proteins,with no Effect on Milk Composition or Yield

IGF-I regulates lactation by stimulating mammary mitogenesis, inhibiting apoptosis, and partially mediating the effects of growth hormone on lactogenesis. Herein, lactation performance during first and second parity was assessed in transgenic swine (TG) that over-expressed human IGF-I in milk under the control of the bovine α-lactalbumin promoter, regulatory regions and signal peptide coding sequence. Milk samples were collected throughout lactation (farrowing to d24) from TG sows and non-transgenic littermates (CON) and IGF-I, IGF-II, and IGFBP determined. Colostral (<24 h postpartum) IGF-I content was 26-fold greater (p < 0.001) in TG sows (949 ± 107 μg/L; range 228–1600 μg/L) than CON (36 ± 17.8 μg/L) and was 50- to 90-fold greater (p < 0.001) in mature milk (d2-24 postpartum). There was no effect of parity on milk IGF-I content. Milk IGF-II concentration was unaffected by IGF-I over-expression. Low molecular weight IGFBP (IGFBP-2 and -5) in the milk of TG sows were higher (p = 0.02) than CON in the early postpartum period, but did not differ in mature milk. Milk yield, determined by weigh-suckle-weigh, was similar in TG and CON as was litter weight gain. Milk nutrient composition was not significantly affected by IGF over-expression. Thus, mammary specific transgenic over-expression of IGF-I significantly increased milk IGF-I and IGFBP content, but did not impact lactation performance in swine.     

Circulating levels of α‐tocopherol and retinol in free‐ranging African elephants (Loxodonta africana)

To date, there are no detailed reports of circulating levels of plasma α‐tocopherol and retinol for large samples of free‐ranging African elephants (Loxodonta africana). This survey study measured natural circulating levels of α‐tocopherol as a measure of vitamin E activity and retinol as an indicator of vitamin A activity, in 70 free‐ranging African elephants captured at Kruger National Park as part of a translocation program. Mean levels of α‐tocopherol and retinol were found to be 0.613 ± 0.271 μg/mL and 0.039± 0.007 μg/mL, respectively, and did not vary significantly across sex or age class. Elephants appear to normally have low circulating levels of both these nutrients compared with domestic herbivore species; values from healthy, free‐ranging elephant populations may provide useful data for assessing nutrient status of captive animals. Zoo Biol 18:319–323, 1999.© 1999 Wiley‐Liss, Inc.     

Properties of liver retinyl ester hydrolase in young pigs

A new sensitive method for the assay of retinyl ester hydrolase in vitro was developed and applied to liver homogenates of 18 young pigs with depleted-to-adequate liver vitamin A reserves. Radioactive substrate was not required, because the formation of retinol could be adequately quantitated by reversed-phase high-performance liquid chromatography. Optimal hydrolase activity was observed with 500 μM retinyl palmitate, 100 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, and 2 mg/ml Triton X-100 at pH 8.0. The relative rates of hydrolysis of six different retinyl esters by liver homogenate were: retinyl linolenate (100%), myristate (99%), palmitate (47%), oleate (38%), linoleate (31%), and streate (29%). The enzyme was found primarily in the membrane-containing fractions of liver (59±3%, S.E.) and kidney (76±3%), with considerably lower overall activity in kidney (57–375 nmol/h per g of tissue) than in liver (394–1040 nmol/h per g). Retinyl ester hydrolase activity in these pigs was independent of serum retinol values, which ranged from 3 to 24 μg/dl, and of liver vitamin A concentrations from 0 to 32 μg/g. Pig liver retinyl ester hydrolase from the rat liver enzyme in its substrate specificity, bile acid stimulation, and interanimal variability.     

High-protein diet during gestation and lactation affects mammary gland mRNA abundance, milk composition and pre-weaning litter growth in mice

We evaluated the effect of a high-protein diet (HP) on pregnancy, lactational and rearing success in mice. At the time of mating, females were randomly assigned to isoenergetic diets with HP (40% w/w) or control protein levels (C; 20%). After parturition, half of the dams were fed the other diet throughout lactation resulting in four dietary groups: CC (C diet during gestation and lactation), CHP (C diet during gestation and HP diet during lactation), HPC (HP diet during gestation and C diet during lactation) and HPHP (HP diet during gestation and lactation). Maternal and offspring body mass was monitored. Measurements of maternal mammary gland (MG), kidney and abdominal fat pad masses, MG histology and MG mRNA abundance, as well as milk composition were taken at selected time points. HP diet decreased abdominal fat and increased kidney mass of lactating dams. Litter mass at birth was lower in HP than in C dams (14.8 v. 16.8 g). Dams fed an HP diet during lactation showed 5% less food intake (10.4 v. 10.9 g/day) and lower body and MG mass. On day 14 of lactation, the proportion of MG parenchyma was lower in dams fed an HP diet during gestation as compared to dams fed a C diet (64.8% v. 75.8%). Abundance of MG α-lactalbumin, β-casein, whey acidic protein, xanthine oxidoreductase mRNA at mid-lactation was decreased in all groups receiving an HP diet either during gestation and/or lactation. Milk lactose content was lower in dams fed an HP diet during lactation compared to dams fed a C diet (1.6% v. 2.0%). On days 14, 18 and 21 of lactation total litter mass was lower in litters of dams fed an HP diet during lactation, and the pups' relative kidney mass was greater than in litters suckled by dams receiving a C diet. These findings indicate that excess protein intake in reproducing mice has adverse effects on offspring early in their postnatal growth as a consequence of impaired lactational function.     

Radioimmunoassay of androsterone and androsterone-3-sulfate in plasma

Details of a sensitive and specific radioimmunoassay for androsterone (1) and androsterone sulfate in plasma have been presented. Benzene extracts of plasma were chromatographed on a lumina to isolate the androsterone fraction either (a) directly after extraction (A) or (b) after solvolysis (AS). Following treatment with rabbit anti-A-17-BSA, antibody bound steroid was precipitated by ammonium sulfate. Androsterone concentrations in normal male plasma averaged 57 ± 24 (S.D.) ng/dl, range 35–135 ng/dl and for normal women, 44 ± 21 (S.D.) ng/dl, range 18–98 ng/dl. Androsterone sulfate concentrations were: males 55 ± 28 μg/dl (range 10–114 μg/dl); premenopausal females 52 ± 31 μg/dl (range 16–318 μg/dl).     

Ultrafiltration-light absorption spectrophotometric determination of silicon in blood

An ultrafiltration-light absorption spectrometric method for soluble molybdate-reactive silicon was assessed and applied to bovine and ovine blood plasma and sera, giving precise analytical results. Interfering protein above molecular weight 10,000–25,000 was removed by ultrafiltration, and silicon in ultrafiltrates was quantitated by measuring light absorption at 810 nm of the 1,2,4-aminonaphthol sulfonic acid/ascorbic acid-reduced silicomolybdate. Chemical interferences on the color-forming reaction of remaining blood components were tested by measuring recoveries of silicon added to real blood plasma samples and to synthetic blood plasma solutions, the latter containing typical levels of the major ions Na⁺, K⁺, Ca²⁺, HCO₃^?, and Cl^?, together with varying quantities of the potential interferants (amount per analytical reaction): phosphate (0–0.5 mg P), ferric ion (0–3 mg), fluoride (0–1.25 mg), vanadate (0–0.5 mg V), arsenate (0–10 μg As), and germanate (0–0.5 μg Ge). The mean recovery of added 0.8–9 μg silicon/g of bovine and ovine plasma was 97.7% (SE = 1.0, n = 17); the mean recovery of 1 and 5 μg silicon from synthetic blood plasma solutions with interferant levels up to 50-fold that in normal plasma was 99.2% (SE = 0.3, n = 47). Silicon concentrations found in bovine and ovine blood plasma and sera were typically around 7 μg/ml with procedural reagent blanks consistently low at a mean of 0.12 μg/test (SD = 0.011, n = 20). The silicon level in Center for Disease Control bovine serum (reference specimen Lot R-2274) was found to be (mean ± SE, n = 10) 1.147 ± 0.013 μg/g or 1.172 ± 0.013 μg/ml (25°C). The method detectivity (detection limit) was estimated at 0.03 μg.     

Free-radical scavenging by Ouratea parviflora in experimentally-induced liver injuries

Abstract

The antioxidant potential of crude extracts and fractions from leaves of Ouratea parviflora, a Brazilian medicinal plant used for the treatment of inflammatory diseases, was investigated in vitro through the scavenging of radicals 2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), hydroxyl radical (HO^?), superoxide anion (O₂^??), and lipid peroxidation in rat liver homogenate. The crude extract (CEOP) and hydro-alcoholic fraction (OP4) showed strong inhibitory activity toward lipid peroxidation induced by tert-butyl peroxide (IC₅₀ = 2.3 ± 0.2 and 1.9 ± 0.1 μg/ml, respectively). The same products exhibited a strong concentration-dependent inhibition of deoxyribose oxidation (14.9 ± 0.2 and 0.2 ± 0.1 μg/ml, respectively), and also showed a considerable antioxidant activity against O₂^??(87.3 ± 0.1 and 73.1 ± 0.4 μg/ml, respectively) and DPPH radicals (55.4 ± 0.3 and 38.3 ± 0.4 μg/ml, respectively). The protective effects of CEOP and OP4 were also studied in mouse liver. CCl₄ significantly increased (by 90%) levels of lipid hydroperoxides, carbonyl protein content (64%), DNA damage index (133%), aspartate aminotransferase (261%), alanine aminotransferase (212%), catalase activity (23%), and also caused a decrease of 60% in GSH content. The results showed that CEOP and OP4 exerted cytoprotective effects against oxidative injury caused by CCl₄ in rat liver, probably related to the antioxidant activity showed by the in vitro free radical scavenging property.