Health benefits of orally administered anti-IL-10 antibody in milk-fed dairy calves.

The primary objective of this randomized controlled trial was to determine whether anti-IL-10 egg yolk antibodies fed upon arrival to a calf ranch would lower the prevalence of Cryptosporidium parvum shedding in naturally challenged preweaned dairy calves. The secondary objectives included measuring the effect of anti-IL-10 antibodies on calf health, performance, and shedding of less common diarrheal pathogens. A total of 133 calves, enrolled at 24 to 72 h of age, received a daily dose of 0.96 g of egg yolk powder with anti-IL-10 antibodies (MAB, n = 71) or without anti-IL-10 antibodies (MEP, n = 62) split between 2 feedings for the first 11 d on feed at a calf ranch. Daily health evaluations were completed for 15 d after arrival and on d 56. Digital weights were collected at enrollment and d 56, and hipometer weights were collected at enrollment and d 7 and 56. Packed cell volume and serum total protein concentration were measured at enrollment and on d 7 and 14. Fecal pH was measured at enrollment and on d 5 and 14, and fecal pathogen (C. parvum, coronavirus, rotavirus, and Salmonella spp.) shedding was assessed at d 5 and 14. Continuous outcomes were compared between groups using a Student's t-test or Wilcoxon rank sum test. Fecal pathogen shedding at d 14, respiratory disease at d 56, and antibiotic usage were compared using relative risk (RR) and chi-squared test. Fecal pH (median and interquartile range) on d 14 was 6.65 (6.39-6.99) and 6.52 (5.97-6.81) for MAB and MEP, respectively. On d 56, the risk of respiratory disease was lower for MAB compared with MEP (RR = 0.40; confidence interval = 0.16-0.99). The risk for antibiotic treatment was lower for MAB- compared with MEP-treated calves (RR = 0.38; confidence interval = 0.17-0.88). The risk of shedding rotavirus was higher in MAB (RR = 1.38; confidence interval = 1.10-1.81) calves. After multivariable analyses, hipometer weights (least squares means ± standard error) were 1.7 ± 0.8 kg greater on d 56 in MAB compared with MEP; however, ADG was 0.04 ± 0.02 kg/d lower in MAB calves. Total health score, diarrhea days, average respiratory score, packed cell volume, and serum total protein were not affected by feeding anti-IL-10 egg antibodies. In summary, feeding anti-IL-10 antibodies was associated with increased fecal pH, reduced risk of respiratory disease later in the preweaning period, and decreased antibiotic usage despite higher rotavirus infection. These findings might be associated with improved mucosal immunity, enhanced host defenses, or reduced susceptibility and warrant further investigation.

Breath carbon stable isotope ratios identify changes in energy balance and substrate utilization in humans.

Rapid detection of shifts in substrate utilization and energy balance would provide a compelling biofeedback tool for individuals attempting weight loss. As a proof of concept, we tested whether the natural abundance of exhaled carbon stable isotope ratios (breath δ(13)C) reflects shifts between negative and positive energy balance. Volunteers (n=5) consumed a 40% energy-restricted diet for 6 days followed by 50% excess on day 7. Breath was sampled immediately before and 1 h and 2 h after breakfast, lunch and dinner. Exhaled breath δ(13)C values were measured by cavity ring-down spectroscopy. Using repeated measures analysis of variance (ANOVA) followed by Dunnett's contrasts, pre-breakfast breath values on days 2-6 were compared with day 1, and postprandial day 7 time points were compared with pre-breakfast day 7. Energy restriction diminished pre-breakfast breath δ(13)C by day 3 (P<0.05). On day 7, increased energy intake was first detected immediately before dinner (-23.8±0.6 vs -21.9±0.7‰, P=0.002 (means±s.d.)), and breath δ(13)C remained elevated at least 2 h post dinner. In conclusion, when shifting between negative and positive energy balance, breath δ(13)C showed anticipated isotopic changes. Although additional research is needed to determine specificity and repeatability, this method may provide a biomarker for marked increases in caloric intake.

ß-Cryptoxanthin biofortified maize (Zea mays) increases ß-cryptoxanthin concentration and enhances the color of chicken egg yolk.

The laying hen has a natural ability to deposit carotenoids into its egg yolks, especially the xanthophyll carotenoid lutein that is used commercially as an egg colorant. Can this ability to deposit carotenoids be used to enrich egg yolk provitamin A value? After a 10-d carotenoid depletion period in hens (n = 24), the effects of a 20-d intervention with high-ß-cryptoxanthin, high-ß-carotene, or typical yellow maize on color and carotenoid profile were compared with the effects of a white maize diet (n = 6/treatment). Eggs were collected every other day and yolks were analyzed by using a portable colorimeter to define the color space and by using an HPLC to determine the carotenoid profile. The high-ß-cryptoxanthin and yellow maize increased ß-cryptoxanthin in the yolk (0.55 ± 0.08 to 4.20 ± 0.56 nmol/g and 0.55 ± 0.08 to 1.06 ± 0.12 nmol/g, respectively; P < 0.001). Provitamin A equivalents increased in eggs from hens fed high-ß-cryptoxanthin maize (P < 0.001) but not the high-ß-carotene maize. The color (L*, a*, and b*) assessment of the yolks showed an increase in the high-ß-cryptoxanthin treatment for the red-green a* scale (P < 0.001) and a decrease for the light-dark L* scale (P < 0.001). No appreciable change was noted in the yellow-blue b* scale for the high-ß-cryptoxanthin treatment; but significant changes were noted for the yellow (P = 0.002) and high-ß-carotene maize (P = 0.005) treatments, which were most evident at the end of the washout period with white maize. ß-Cryptoxanthin-biofortified maize is a potential vehicle to elevate provitamin A equivalents and to enhance the color of yolks. This could lead to a human health benefit if widely adopted.

Evidence that maternal conjugated linoleic acid negatively affects lipid uptake in late-stage chick embryos resulting in increased embryonic mortality.

An experiment was performed to determine the effect of maternal dietary conjugated linoleic acid (CLA) on growth and composition of surviving chick embryos and residual yolk sacs during the last week of development when lipid utilization becomes prevalent. After 14 d on experimental diets, hatchability of non-cooled eggs obtained from CLA-fed hens (0.5% of the diet) was 10%, where 20% of surviving CLA embryos died after d 13 of incubation. Hatchability was 93% for controls and only 4.36% of mortality occurred after d 13 of incubation. Decline in yolk sac weight in control embryos (0.75 g/d) was significantly greater than that from viable CLA embryos (0.51 g/d). Growth rate (2.6 g/d) of surviving embryos from d 13 to 20 was reduced in CLA embryos in comparison to growth rate of controls (3.0 g/d). Relative proportion of lipid in residual yolk sacs in embryos from control-fed hens decreased from 26.72% (d 13) to 15.94% (d 19) during incubation, whereas little change was evident in residual yolk sac from CLA embryos on d 13 (21.52%) to d 19 (20.39%). Fatty acid analysis of residual yolk sac contents suggested that transport of fatty acids from the contents (liquid yolk) to the yolk sac membrane was not impaired in CLA embryos, as shown by a similar pattern in reduction of total fatty acids in residual yolk sac contents between treatment groups. Apart from 18:1n-9 (d 17), there were no consistent differences in the fatty acid content between embryos from hens fed the control diet or the CLA diet at any time point. Maternal CLA led to increased 18:0 and decreased 18:1n-9 in yolk lipid and embryonic tissues compared with controls across time. These findings could possibly suggest that CLA embryos had less capacity to use yolk lipids from the residual yolk sac during the last week of incubation.

Egg yolk antibodies for detection and neutralization of Clostridium botulinum type A neurotoxin.

The objective of this research project was to determine the usefulness of an egg antibody platform for producing materials for the detection and neutralization of botulinum type A neurotoxin. Yield estimates for detection and neutralizing antibodies produced using methods described were calculated. Antibody specific to botulinum toxoid A (aToxoid) and toxin A (aBoNT/A) was produced by immunizing hens with botulinum toxoid A (toxoid) followed by increasing amounts of botulinum neurotoxin A (BoNT/A) in Freund incomplete adjuvant. Egg yolks were extracted with polyethylene glycol (PEG) for antibody detection and neutralization experiments. A model aToxoid/toxoid immunoassay using only egg yolk antibody was developed and had a detection limit of 1 pg/ml of toxoid. In an indirect enzyme-linked immunosorbent assay of BoNT/A-specific antibody, the aBoNT/A contained more BoNT/A-specific antibody than did the aToxoid, and aBoNT/A was as effective as commercial rabbit antibody. The aToxoid provided no protection against BoNT/A in a standard mouse neutralization assay; however, 1 mg of PEG-extracted aBoNT/A neutralized 4,000 lethal doses of BoNT/A injected intraperitoneally. Based on these results, we calculated that in 1 month one hen could produce more than 100 liters of antibody detection reagents or enough antibody to neutralize approximately 11.6 million mouse lethal doses of botulinum toxin. Utilization of an egg antibody platform is potentially rapid (28 to 70 days) and scalable to kilogram quantities using current egg production facilities with as few as 1,000 hens.

Evidence for conjugated linoleic acid-induced embryonic mortality that is independent of egg storage conditions and changes in egg relative fatty acids.

Three experiments were performed to determine the effect of conjugated linoleic acid (CLA) on embryonic development in the absence of vitelline membrane disruption. In experiment 1, when eggs from control and CLA (0.5%)-fed hens were stored at 21 or 15 degrees C for 48 h, mineral movement between the yolk and albumen was not observed (with the exception of Mg and Na). Also, it was found that CLA-induced changes in yolk fatty acid content (e.g., increased saturated fatty acids and CLA) had begun to change after 5 d of feeding hens CLA, and no differences were detected in fatty acid composition after 14 d. In experiment 2, the hatchability of eggs incubated directly after oviposition or stored 24 h at 21 or 15 degrees C was determined from hens fed control or 0.5% CLA diets. Regardless of storage conditions, CLA reduced hatchability. These data showed that CLA elicits negative effects on hatchability independent of vitelline membrane disruption or egg storage condition. In experiment 3, eggs were collected from hens fed 0 or 1% CLA daily for 3 wk, stored at 21 degrees C for 24 h, and incubated. Not only did CLA decrease hatchability, the data showed as the days of CLA feeding increased, the days of survival during incubation decreased. Average days of embryonic survival during incubation for the CLA group diminished to 18.0, 13.4, and 6.3 d for wk 1, 2, and 3 of CLA feeding, respectively, and control remained at 20.6, 20.8, and 19.8 for the 3 wk. These studies suggested that without the disruption of the vitelline membrane, hatchability and embryonic days of survival were significantly reduced by maternal CLA feeding in comparison to control-fed hens. Evidence that embryos die earlier the longer the hens are fed CLA, even though no additional changes in the fatty acid content of eggs were found, suggested that factors other than storage and egg yolk fatty acid composition played a role in CLA-induced embryonic mortality.

Carrot leaves improve color and xanthophyll content of egg yolk in laying hens but are not as effective as commercially available marigold fortificant1.

Yellow and orange egg yolks are good sources of xanthophyll carotenoids, consumption of which is associated with health benefits, such as cancer prevention, eye health, and bone health. Industrial feed fortificants used to improve egg yolk color and carotenoid concentration typically are derived from marigold flowers. Green leafy vegetables are also concentrated sources of the xanthophylls lutein and zeaxanthin (L+Z), but they have not been rigorously evaluated in laying hen feeds as a yolk colorant. The addition of food manufacturing byproducts, including carrot leaves, to animal feed is a promoted method of improving animal nutrition. The ability of dehydrated carrot leaves to improve egg yolk color and L+Z concentration was evaluated by feeding laying hens (n = 40) white maize-based feeds fortified with 2 different dehydrated carrot leaves, marigold as a positive control, or no fortificant as a negative control for 28 D. After a 7-D washout period, the hens were separated into 4 groups, and eggs were collected every other day. Yolks were analyzed by using a portable colorimeter to define the color space and by ultra-performance liquid chromatography to determine the carotenoid profile. Carotenoid concentration rapidly declined from day 0 to 8, confirming adequate washout conditions. The white maize negative control (WM) day 28 lutein concentration (3.59 ± 0.51 nmol/g) was significantly less than orange-carrot leaf-treated (OCL) (5.34 ± 0.36 nmol/g) and red-carrot leaf-treated hens (RCL) (5.92 ± 1.00 nmol/g) in addition to the marigold-treated hens (MG). However, MG was significantly higher than both leaf-treated groups. From day 8 (3.93 ± 0.74 nmol/g) to 28 (9.32 ± 1.66 nmol/g), MG had the largest increase in lutein and was the only treatment to surpass day 0 initial concentrations (8.50 ± 1.64 nmol/g). A similar trend was observed for zeaxanthin and was reflected in the color space.

Protein and energy intake improved by in-between meals: An intervention study in hospitalized patients.

BACKGROUND/AIM: Disease related malnutrition is a major problem in hospitals. Malnutrition in hospitalized patients is caused by many factors. Among these factors are decreased appetite and early satiety, and reaching nutritional requirements in nutritional risk patients is a challenge when using ordinary energy and protein dense food. The aim of this study was to examine if total protein and energy intake in medical and surgical patients at nutritional risk could be improved by protein fortified and energy rich in-between meals. METHODS: An assortment of fortified in-between meals including 10 g of protein was developed based on patient preferences and served in the Departments of Lung Medicine and Abdominal Surgery for a period of three months. Nutrition intake was recorded before and after intervention. RESULTS: Food intake records were collected from a total of 92 patients, (46 before and 46 after intervention). The total amount of protein intake per in-between meal was increased from 2,6 g to 10,3 g. Total daily protein intake increased from 49% to 88% (p < 0.00) and total energy intake from 74% to 109% (p < 0.00) of requirements. CONCLUSION: Protein and energy intake for surgical and medical patients at in-between meals as well as total daily intake increased significantly. Recommended average level for individually measured requirements was reached.

Additions of killed whole cell bacteria preparations to Freund complete adjuvant alter laying hen antibody response to soluble protein antigen.

Passive transfer of antibodies from hen to egg has value to both the producer of commercial polyclonal egg antibody and the producer of hatching eggs. Water-in-oil emulsions are commonly amended with immune stimulants such as Mycobacteria (e.g., Freund complete adjuvant; FCA) to increase antibody production to soluble protein antigens (SPA). Recent discoveries of the mechanisms by which microbial products act as adjuvants led us to hypothesize that additions of killed whole cell bacteria (bacterins) to FCA could improve antibody responses to SPA. All injections used in each experiment were water-in-oil emulsions (50:50) containing 3 mg/mL of phospholipase A(2) (PLA(2)) immunogen. Additionally, all primary control and treatment injections contained heat-killed Mycobacterium butyricum immunogens from FCA. In addition to PLA(2) and FCA, primary treatment injections contained various microbial bacterin immunogens. Hence, the experimental treatment of all experiments was addition of a commercial source of microbial bacterin to FCA for the primary injection only. Booster injections were the same as the primary control injections except Freund incomplete adjuvant replaced FCA. Anti-body titers to PLA(2) in yolk were determined by ELISA. Bacterins tested as additives to FCA were Escherichia coli, Staphylococcus aureus, Streptococcus suis, and Corynebacterium pseudotuberculosis. Escherichia coli bacterin added to FCA decreased egg yolk antibody titer to SPA by 23% in hens of different ages and strains (P < 0.0001). In a second experiment, a 51% decrease in antibody production associated with E. coli bacterin was sustained for several weeks after the primary immunization (P = 0.003). Staphylococcus aureus or Streptococcus suis combined with FCA increased egg yolk antibody 62 and 51%, respectively (P < 0.05), and Corynebacterium pseudotuberculosis had no effect. In conclusion, the addition of bacterin to FCA can influence hen antibody response to SPA as measured in egg yolks. It is hypothesized that the difference in antibody production may be related to the composition of various pathogen associated molecular patterns in the primary injection.

Vaccine to fibroblast growth factor 23 peptides increases eggshell strength.

Strategies that would increase eggshell quality could be of considerable value to egg producers. This research demonstrated the effective use of fibroblast growth factor 23 (FGF-23) peptide vaccines to increase eggshell quality of Single Comb White Leghorn laying hens (from 69 to 72 wk of age). Hens, fed a standard diet (containing 900 IU/kg vitamin D3), were intramuscularly injected (and boosted) with either a control vaccine (n = 14 hens) or one of 2 FGF-23 peptide vaccines (peptides NP1, GMNPPPYS; and NP7, YTSTERNSFH; n = 15 hens for each peptide). During peak antibody titer, eggs were collected for shell and internal quality analysis, hens were artificially inseminated, and the hatchability of fertilized eggs was determined. Laying hens vaccinated with either FGF-23 peptide NP1 or NP7 had increased (P < 0.05) plasma phosphate level (mmol/L; NP1 = 1.74, NP7 = 1.76, control = 1.47), egg specific gravity (NP1 = 1.083, NP7 = 1.083, control = 1.079), and eggshell strength (g of force; NP1 = 4002, NP7 = 4157, control = 3102) when compared to control vaccinated hens. FGF-23 peptide NP1 vaccinated hens also had increased eggshell thickness (mm, P < 0.001), shell weight (g, P = 0.032), and shell index (% of whole egg, P = 0.023) when compared to control vaccinated hens. FGF-23 peptide NP7 vaccinated hens tended to have decreased eggshell weight (P = 0.064) when compared to control vaccinated hens. Hatchability of fertilized eggs was not affected in incubations 1 and 3, but tended to be decreased (P = 0.097) by FGF-23 peptide NP1 vaccination in incubation 2. In conclusion, vaccines to FGF-23 peptides increased eggshell quality of laying hens with minimal adverse effects on egg internal quality. The effect of FGF-23 peptide vaccination on hatchability remains to be clarified.

Regulation of beta 1-integrin-mediated cell adhesion by the Cbl adaptor protein.

BACKGROUND: Leukocyte activation results in a rapid increase in adhesion to the extracellular matrix due to the activation of beta 1 integrin receptors. A role for phosphatidylinositol (PI) 3-kinase in integrin activation has been proposed, as activation of integrins by many receptors can be blocked by PI 3-kinase inhibitors. One receptor that regulates integrins is the CD28 surface antigen; here, we investigated the mechanisms responsible for CD28-mediated integrin activation. RESULTS: CD28-mediated integrin activation was blocked by mutation of the binding site for the p85 catalytic subunit of PI 3-kinase in the CD28 cytoplasmic domain, and by expression of a dominant-negative form of the p85 subunit. Substitution of the Src homology 2 (SH2)-binding motif in the CD28 cytoplasmic domain for the corresponding motif in the CD28-related CTLA-4 surface antigen also blocked integrin activation but did not affect the recruitment and activation of PI 3-kinase. Mutations of the CD28 cytoplasmic domain that blocked integrin activation also impaired the tyrosine phosphorylation of the Cbl adaptor protein and the activation of the PI 3-kinase that was associated with Cbl. This Cbl-associated PI 3-kinase was distinct from the PI 3-kinase that coprecipitated with the CD28 cytoplasmic domain. CD28-mediated activation of beta 1 integrins was inhibited by expression of a mutant Cbl protein that shows reduced association with PI 3-kinase. CONCLUSIONS: Cbl is required for PI-3-kinase-dependent regulation of integrin receptors by CD28. Furthermore, CD28 is coupled to two distinct pools of PI 3-kinase, one directly associated with the CD28 cytoplasmic tail and the other associated with Cbl.

Additive effects of fibroblast growth factor 23 neutralization and dietary phytase on chick calcium and phosphorus metabolism.

Phytase hydrolyzes phytate rendering phosphorus available for intestinal absorption, while systemic neutralization of fibroblast growth factor 23 (FGF-23), using anti-FGF-23 antibody, has been shown to increase phosphate retention. Hence, neutralization of FGF-23 should be additive with phytase in reducing dietary non-phytate phosphorus (nPP) needs in chickens fed plant-based diets rich in phytic acid. This study was designed to test the additive effects of maternally derived anti-FGF-23 antibody and dietary phytase on the performance of chicks fed a low nPP diet from one to 14 d. Single Comb White Leghorn laying hens were vaccinated with either an adjuvant control or a synthetic FGF-23 peptide (GMNPPPYS). Chicks from vaccinated hens with control or anti-FGF-23 maternal antibodies were fed either a diet containing 0.2% nPP and 0.9% calcium with or without 500 unit phytase per kg of diet (2 × 2 factorial with main effects of antibody type and phytase addition, n = 15 pens of chicks/treatment). A significant interaction between dietary phytase and maternally derived anti-FGF-23 antibody on growth and feed efficiency was observed (P ≤ 0.05), in which chicks receiving either phytase or maternally derived anti-FGF-23 antibody had improved body weight gain (21 or 15%, respectively) and feed efficiency (16 or 18%, respectively) as compared to chicks with control antibody and not fed phytase. Both phytase and maternally derived anti-FGF-23 antibody independently increased (P ≤ 0.05) plasma phosphate (11 and 11%, respectively) and percent tibiotarsus ash (13 and 11%, respectively). Significant main effects and the lack of an interaction supported an additive effect of phytase and anti-FGF-23 antibody on plasma phosphate and percent tibiotarsus ash. Feeding phytase to chicks fed 0.2% nPP increased plasma FGF-23 levels by 22% (P ≤ 0.05); however, no effects of anti-FGF-23 antibody on plasma FGF-23 levels were observed. In conclusion, dietary phytase and presence of anti-FGF-23 antibody have an additive effect on plasma phosphate and tibiotarsus ash in chicks fed low nPP diets. Data support that phytase and anti-FGF-23 antibody increase phosphate utilization by different mechanisms.

The biologically active isomers of conjugated linoleic acid.

Numerous physiological effects are attributed to conjugated linoleic acid (CLA). The purpose of this presentation is to consider these effects with respect to the cis-9,trans-11 and trans-10,cis-12 CLA isomers. We review previously published data and present new findings that relate to underlying biochemical mechanisms of action. Both isomers are natural products. The cis-9,trans-11 isomer is the principal dietary form of CLA, but the concentrations of this isomer and the trans-10,cis-12 isomer in dairy products or beef vary depending on the diet fed to cows or steers, respectively. The trans-10,cis-12 CLA isomer exerts specific effects on adipocytes, in particular reducing the uptake of lipid by inhibiting the activities of lipoprotein lipase and stearoyl-CoA desaturase. The trans-10,cis-12 CLA isomer also affects lipid metabolism in cultured Hep-G2 human liver cells, whereas both the cis-9,trans-11 and trans-10,cis-12 CLA isomers appear to be active in inhibiting carcinogenesis in animal models. We present new findings indicating that the cis-9,trans-11 CLA isomer enhances growth and probably feed efficiency in young rodents. Accordingly, the effects of CLA on body composition (induced by trans-10,cis-12 CLA) and growth/feed efficiency (induced by cis-9,trans-11 CLA) appear to be due to separate biochemical mechanisms. We also show that a 19-carbon CLA cognate (conjugated nonadecadienoic acid, CNA) inhibits lipoprotein lipase activity as effectively as CLA in cultured 3T3-L1 adipocytes. Presumably, CNA is metabolized differently than the 18-carbon CLA isomers, so this finding indicates direct activity of the administered compound as opposed to acting via a metabolite.

Host-targeted approaches to managing animal health: old problems and new tools.

Our fellow medical and regulatory scientists question the animal producer's dependence on antibiotics and antimicrobial chemicals in the production of animal products. Retail distributors and consumers are putting even more pressure on the animal industry to find new ways to produce meat without antibiotics and chemicals. In addition, federal funding agencies are increasingly pressuring researchers to conduct science that has application. In the review that follows, we outline our approach to finding novel ways to improve animal performance and health. We use a strict set of guidelines in our applied research as follows: (1) Does the work have value to society? (2) Does our team have the skills to innovate in the field? (3) Is the product we produce commercially cost-effective? (4) Are there any reasons why the general consumer will reject the technology? (5) Is it safe for the animal, consumer, and the environment? Within this framework, we describe 4 areas of research that have produced useful products, areas that we hope other scientists will likewise explore and innovate such as (1) methods to detect infection in herds and flocks, (2) methods to control systemic and mucosal inflammation, (3) improvements to intestinal barrier function, and (4) methods to strategically potentiate immune defense. We recognize that others are working in these areas, using different strategies, but believe our examples will illustrate the vast opportunity for research and innovation in a world without antibiotics. Animal scientists have been given a new challenge that may help shape the future of both animal and human medicine.

The presence of lysylpyridinoline in the hypertrophic cartilage of newly hatched chicks.

The presence of lysylpyridinoline (LP) as a nonreducible cross-link in appreciable quantities has primarily been limited to the mineralized tissues, bone and dentin. However, the results reported here show that LP is not only present in the hypertrophic cartilage of the tibiotarsus isolated from newly hatched broiler chicks, but it is approx. 4-fold as concentrated as hydroxylysylpyridinoline (HP). Bone and articular cartilage surrounding the hypertrophic cartilage do not contain measurable quantities of LP. Purified LP has a fluorescent scan similar to purified HP and literature values, confirming that we indeed were measuring LP. Also, the cartilage lesion produced by immature chondrocytes from birds with tibial dyschondroplasia had LP but the HP:LP ratio was > 1. Thus, the low HP:LP ratio could be a marker for hypertrophic cartilage in avians.

Inhibition of hepatic stearoyl-CoA desaturase activity by trans-10, cis-12 conjugated linoleic acid and its derivatives.

Conjugated linoleic acid (CLA) has been reported to decrease stearoyl-CoA desaturase (SCD) activity by decreasing mRNA expression. This investigation was designed to determine whether structurally related compounds of CLA have a direct inhibitory effect on SCD activity. Trans-10,cis-12 CLA had strong inhibitory activity on SCD while cis-9,trans-11, and trans-9,trans-11 isomers had no effect. Trans-10 octadecenoate was not inhibitory, whereas cis-12 octadecenate was inhibitory, but not as effective as trans-10,cis-12 CLA. Of the oxygenated derivatives, 9-peroxy-cis/trans-10, trans-12 octadecadienoate was a more effective inhibitor than trans-10,cis-12 CLA, whereas 9-hydroxy-trans-10, cis-12 octadecadienoate was less effective. Interestingly, cis-11 octadecadienoate and cis-12 octadecen-10-ynoate were slightly inhibitory. However, trans-9 and trans-11 octadecenoates, and trans-9,cis-12 octadecadienoate were all inactive under test condition, as were linoleate, oleate, and arachidonate. Derivatives of CLA acid modified to alcohol, amide or chloride were all inactive. A cis-12 double bond appears to be a key structural feature for inhibiting SCD activity, especially when coupled with a trans-10 double, whereas a cis-11 double bond is less effective.

Intravenous dilevalol. Effects of the R-R optical isomer of labetalol in patients with severe hypertension.

Dilevalol hydrochloride, the R-R optical isomer of labetalol hydrochloride, was administered intravenously to subjects with severe hypertension. Twelve subjects with supine diastolic blood pressure of more than 115 mm Hg (mean, 124 +/- 2 mm Hg) were studied. Initial doses of 25 mg of dilevalol administered as a slowly given bolus reduced blood pressure by 14/16 mm Hg. With subsequent additional boluses to a total dose of up to 600 mg, supine diastolic blood pressure was reduced to less than 95 mm Hg in ten of 12 subjects studied (mean reduction, 28 mm Hg). Side effects were minimal and upright blood pressure at the time of reduction of blood pressure to goal was not significantly different from supine blood pressure. Plasma renin activity decreased in 11 of 11 subjects studied. Plasma epinephrine concentrations did not change significantly, whereas plasma norepinephrine concentrations increased 2 1/2-fold, probably reflecting the effect of beta 2-agonism on norepinephrine release. Dilevalol appears to be a safe and effective way of lowering blood pressure short term when intravenous antihypertensive therapy is indicated.

Dilevalol in severe hypertension. A multicenter trial of bolus intravenous dosing.

Dilevalol, the R-R optical isomer of labetalol, a nonselective beta-antagonist with vasodilation from selective beta 2 agonism, was administered in sequential multiple bolus intravenous injections of 10 to 100 mg in total doses ranging from 35 to 585 mg (mean dose, 414 mg) to 101 patients with supine diastolic blood pressures above 120 mm Hg. Mean blood pressure was reduced from 200 (+/- 3)/129 (+/- 1) mm Hg to 149 (+/- 2)/101 (+/- 1) mm Hg, a mean reduction of 51/28 mm Hg. The therapeutic goal was established as a reduction in supine diastolic blood pressure to less than 100 mm Hg or a reduction of at least 30 mm Hg. This was achieved in 62 (61%) of 101 patients, with an additional 7 patients having a final supine diastolic blood pressure of 100 mm Hg. Treatment with dilevalol was less successful in black male patients than in the group at large. There was a tendency for older patients to respond better than younger patients. Prior recent treatment of patients with beta-adrenergic antagonists decreased the effectiveness of the drug. Significant orthostatic hypotension was not noted. Sixty-four patients were transferred to oral dilevalol treatment in combination with a diuretic, and blood pressure in this group averaged 160/100 mm Hg after 1 month of therapy. Dilevalol appears to be a safe and effective drug that can be used intravenously successfully in the majority of patients with severe hypertension and provides an alternative to therapy with other agents. It also is a useful agent for oral treatment of these patients after successful intravenous therapy.

Intravenous nicardipine for the treatment of severe hypertension. A double-blind, placebo-controlled multicenter trial.

A placebo-controlled, double-blind multicenter trial was conducted in 123 patients with severe hypertension to examine the efficacy and safety of intravenously administered nicardipine hydrochloride in controlling blood pressure. Seventy-three patients were initially randomized to receive nicardipine treatment. This group had an initial blood pressure of 213 +/- 3/126 +/- 2 mm Hg. Sixty-seven patients achieved the therapeutic goal (diastolic blood pressure less than or equal to 95 mm Hg; systolic blood pressure less than or equal to 160 mm Hg). Fifty patients were randomized to receive placebo solution. Blood pressure in these patients was 216 +/- 3/125 +/- 2 mm Hg. No patient in this group achieved the therapeutic goal during the "blinded" portion of the study. Forty-four of 49 patients who did not respond to placebo administration responded to subsequent treatment with nicardipine. Patients with end-organ damage were included in the study. These included patients with left ventricular hypertrophy, retinopathy, and renal insufficiency. Patients with and without end-organ damage responded equally well to nicardipine treatment. Serious adverse experiences were infrequent, the most common adverse reaction being headache in 24% of the patients studied.

Noncollagenous (NC1) domain of collagen VII resembles multidomain adhesion proteins involved in tissue-specific organization of extracellular matrix.

Type VII collagen (C7) is a stratified squamous epithelial basement membrane protein composed of three identical alpha chains, each consisting of a 145-kDa amino-terminal noncollagenous (NC1) domain and a 145-kDa carboxyl-terminal collagenous domain. Morphologic and biochemical studies have shown that tissue-specific aggregates of C7 dimers called anchoring fibrils may contribute to epithelial basement membrane organization and adherence by interacting with extracellular matrix (ECM) proteins such as type IV collagen. In this study, we cloned a cDNA encoding most of the NC1 domain of C7. The deduced amino acid sequence revealed motifs characteristic of multidomain ECM proteins that contribute to the tissue-specific organization of ECM including a region of 7 1/2 sequential fibronectin type III (Fn III) homology repeats, a potential collagen-binding region homologous to the A domain of von Willebrand factor (vWf) and an RGD sequence. A purified C7 fusion protein containing these motifs specifically bound to type IV collagen in a functional assay. These results suggest that regions within the NC1 domain of C7 mediate interactions with lamina densa and dermal ECM proteins including type IV collagen. Structural mutations and autoepitopes in these regions may represent mechanisms for the development of defective basement membrane organization and adherence in genetic and autoimmune forms of epidermolysis bullosa.