Developing a strawberry yogurt fortified with marine fish oil.

Fortified dairy products appeal to a wide variety of consumers and have the potential to increase sales in the yogurt industry and help increase intake of long-chain n-3 fatty acids. The objectives of this study were to develop a strawberry yogurt containing microencapsulated salmon oil (MSO; 2% wt/vol) and evaluate its characteristics during 1 mo of storage. Unpurified salmon oil (USO) was purified (PSO) and both USO and PSO were analyzed for peroxide value (PV), anisidine value (AV), total oxidation, free fatty acids (FFA), and moisture content. A stable emulsion was prepared with 7% PSO, 22% gum arabic, 11% maltodextrin, and 60% water. The emulsion was spray-dried to produce MSO. The MSO was added to strawberry-flavored yogurt (SYMSO) before pasteurization and homogenization, and a control (SY) without MSO was produced. Both yogurts were stored for 1 mo at 4°C and we determined the quality characteristics including acidity (pH), syneresis, thiobarbituric acid (TBA), fatty acid methyl ester composition, color, and lactic acid bacteria (LAB) count. The entire experiment was replicated 3 times. Total oxidation (unitless) of USO, PSO, and MSO was calculated to be 20.7±1.26, 10.9±0.1, and 13.4±0.25, respectively. Free fatty acid contents were 1.61±0.19%, 0.59±0.02%, and 0.77±0.02% for USO, PSO, and MSO, respectively. Eicosapentaenoic acid and docosahexaenoic acid were the predominant polyunsaturated fatty acids in MSO and in SYMSO, but neither was detected in SY. Fortification of SY with MSO had no significant effect on yogurt pH or syneresis. A decrease in concentration of lactic acid bacteria was observed during the storage of all yogurts. Thiobarbituric acid values significantly increased as storage time increased and SY had a significantly lighter (higher L*) and less yellow (lower b*) color than SYMSO. Although some slight differences were observed in the color and oxidation of SYMSO compared with SY, the study demonstrated that SY could be fortified with salmon oil.

Focal expression of insulin-like growth factor I in rat kidney collecting duct.

To address the question of insulin-like growth factor (IGF) I localization and synthesis in kidney, we used two complementary experimental approaches: immunohistochemistry of fixed paraffin-embedded rat kidney sections; and measurement of IGF I mRNA in isolated components of the rat nephron, using a highly sensitive and specific solution hybridization assay. Immunostainable IGF I was localized exclusively to principal cells of cortical and medullary collecting ducts. Administration of growth hormone to hypophysectomized rats for 8 d resulted in enhanced immunohistochemical staining of IGF I within collecting ducts, but no detectable IGF I in other portions of the nephron. The abundance of IGF I mRNA was 7-12-fold higher in isolated papillary collecting ducts than in proximal tubules or glomeruli, and was enriched 10-fold compared with whole kidney. Our data demonstrate colocalization of IGF I and IGF I mRNA in the collecting duct, consistent with focal expression of the IGF I gene at this site.

Fish protein substrates can substitute effectively for poultry by-product meal when incorporated in high-quality senior dog diets.

An experiment was conducted to analytically define several novel fish substrates and determine the effects of feeding diets containing these substrates on total tract nutrient digestibilities and on immune status of senior dogs. The control diet contained poultry by-product meal while test diets contained 20% milt meal (MM), pink salmon hydrolysate (PSH) and white fish meal (WFM) added at the expense of poultry by-product meal. Concentrations of lymphocytes positive for CD3, CD4, CD8 and CD21 cell-surface markers and immunoglobulin concentrations were measured. Gene expression of cytokines tumour necrosis factor (TNF)-, interleukin (IL)-6, interferon (IFN)-, IL-10 and transforming growth factor (TGF)-ß was determined by quantitative real-time polymerase chain reaction. Major compositional differences were noted among fish substrates but apparent nutrient digestibility coefficients and immune indices were not affected by treatment. Fish protein substrates were found to be effective substitutes for poultry by-product meal, providing diets of high nutritive value for senior dogs.

Carcass composition and yield of Alaskan reindeer (Rangifer tarandus tarandus) steers and effects of electrical stimulation applied during field slaughter on meat quality.

Twenty six adult reindeer steers (>3 years old) were used in a study to evaluate the effect of electrical stimulation (ES) on the quality of hot-boned, rapidly frozen shoulder meat and of the striploin (M. longissimus, LD) from carcasses held at +3°C for 48h. Carcass yield and composition was determined from the left carcass half from which the shoulder meat was not removed. The shoulder meat was processed frozen into cubed, sliced or ground products. Proximate composition of the LD, meat color and water-holding capacity were very similar for the ES (n=15) and non-electrical stimulation (NES; n=11) groups. Ultimate pH and shear force values were significantly lower in the ES meat (LD), however a trained sensory panel could not detect differences between the two groups in any of the measured sensory attributes. Consumer preference tests demonstrated that ES increased tenderness in the cubed and sliced products made from field slaughtered reindeer shoulder meat. ES in combination with hot boning and processing of boneless frozen meat can be used in field slaughter systems for reindeer to improve meat quality and to increase the potential value of the carcass.

Human breast cancer cells contain an error-prone DNA replication apparatus.

The mechanisms responsible for creating genetic errors and genomic instability in cancer cells have not been fully defined. Recently, it has been shown that human cells contain a highly organized complex of proteins, termed the DNA synthesome, that is fully competent to carry out all phases of SV40 in vitro DNA replication (J. M. Coll et al, Oncol. Res., 8: 435-447, 1996; L. H. Malkas et al., Biochemistry, 29: 6362-6374, 1990; Y. Wu et al., J. Cell. Biochem., 54: 32-46, 1994; N. Applegren et al., J. Cell. Biochem., 54: 32-46, 1994). DNA replication fidelity analyses of the DNA synthesome derived from malignant and nonmalignant human breast cells demonstrate that the malignant cell synthesome is mutagenic. The decrease in tumor cell replication fidelity was not due to an increased proliferative capacity of the tumor cells or an increase in the synthetic activity of their DNA synthesome. The ratios of insertions, deletions, and mismatches created by the synthesome from malignant and nonmalignant breast cells were essentially identical, despite the greater overall number of mutations made by the breast cancer cell synthesome. These data define, for the first time, a mechanism unique to cancer cells that contributes to the observed increase in genetic mutation in cancer cells.

A unique form of proliferating cell nuclear antigen is present in malignant breast cells.

Despite extensive research efforts to identify unique molecular alterations in breast cancer, to date, no characteristic has emerged that correlates exclusively with malignancy. Recently, it has been shown that the multiprotein DNA replication complex (synthesome) from breast cancer cells has a significantly decreased replication fidelity compared to that of nonmalignant breast cells. Proliferating cell nuclear antigen (PCNA) functions in DNA replication and DNA repair and is a component of the synthesome. Using two-dimensional PAGE analysis, we have identified a novel form of PCNA in malignant breast cells. This unique form is not the result of a genetic alteration, as demonstrated by DNA sequence analysis of the PCNA gene from malignant and nonmalignant breast cells. This novel form is most likely the result of an alteration in the post-translational modification of PCNA in malignant breast cells. These findings are significant in that it is now possible to link changes in the fidelity of DNA replication with a specific alteration of a component of the DNA synthetic apparatus of breast cancer cells. The novel form of PCNA may prove to be a new signature for malignant breast cells.

Interaction of alpha-actinin, filamin and tropomyosin with F-actin.

The abilities of alpha-actinin, filamin and tropomyosin to bind F-actin were examined by cosedimentation experiments. Results indicated that smooth muscle alpha-actinin and filamin can bind to actin filaments simultaneously with little evidence of competition. In contrast, tropomyosin exhibits marked competition with either filamin or alpha-actinin for sites on actin filaments.

Hormonal effects on the immunocytochemical location of 3',5'-cyclic adenosine monophosphate-dependent protein kinase in rat tissues.

Homogeneous preparations of type I and type II regulatory subunits (RI and RII, respectively) of cAMP-dependent protein kinase (cAMP kinase) were utilized as antigens to obtain isozyme specific antisera. Injections of pure catalytic subunit (C) from the type I isozyme resulted in antisera that reacted with C subunit obtained from either isozyme type. Cross-reactivity of the antisera raised against isolated subunits of the kinase was assessed by immunodiffusion analysis and by measuring the cAMP binding and phosphotransferase activities of the subunits after immunoprecipitation. These antisera were used to localize subunits of type I and type II cAMP kinases in rat skeletal muscle, liver, and adrenal by using indirect immunofluorescence and immunoperoxidase techniques. Specificity of the immunofluorescence was shown by absorption of the antisera with pure homologous antigens. In skeletal muscle, both R and C subunits of the type I and type II cAMP kinases were localized in the area of the sarcoplasmic reticulum and in periodic crossbands. Specific fluorescence for these components was observed in both isotropic and anisotropic band regions of the sarcomere. Densitometric determinations of immunoperoxidase staining revealed a larger amount of RI, RII, and C subunits in the isotropic band than in the anisotropic band regions. In liver, C, RI, and RII subunits were distributed both in cytoplasmic and nuclear areas and along plasma membranes of hepatocytes; however, there were qualitative differences observed among these various subcellular sites. With each antiserum, fluorescence was blocked by prior absorption with homologous antigen. After treatment of rats with glucagon, dramatic changes in the relative distribution patterns of C and RII were noted in the nucleus. In the adrenal gland, RI, RII, and C subunits were localized in both cytoplasmic and nuclear areas, and an apparent redistribution of these subunits occurred after treatment of (dexamethasone-suppressed) rats with ACTH. The application of this immunocytochemical approach provides a tool for examining and monitoring the subcellular distribution of these components of cAMP kinase in biological systems.

Acetylation polymorphism expression in patients before and after liver transplantation: influence of host/graft genotypes.

The consequences of liver transplantation on NAT2 activity were studied in 58 patients of Caucasian origin and compared with a group control of 119 unrelated healthy individuals of the same ethnic origin. Acetylation phenotypes were determined using caffeine as a probe drug before and repeatedly after liver transplantation. NAT2 genotypes were determined with three separate polymerase chain reactions to detect either the NAT2*4 wild-type allele or the NAT2*5A, NAT2*6A and NAT2*7A mutated alleles, associated with a decrease in NAT2 enzyme activity. In patients, the molar urinary elimination ratio AFMU/(AFMU+1X+1U) appeared more reliable than AFMU/1X for assessing the acetylation phenotype and fitted better with the various haplotypes. The variation of xanthine oxidase activity as measured by the 1U/1X urinary elimination ratio, appeared to be responsible for the poor phenotype prediction from the AFMU/1X ratio in post-transplanted patients. Regardless of the pathologic conditions of the treatment in progress, the genotype of the liver played an overwhelming role in the phenotypic expression of NAT2 compared with the genotype of other organs, where NAT2 was expressed in patients who presented a chimerism after liver transplantation.

The influence of insulin-dependent diabetes on the metabolism of caffeine and the expression of the debrisoquin oxidation phenotype.

Twenty-two unrelated healthy subjects and 28 unrelated patients with insulin-dependent diabetes were given 200 mg of caffeine and 10 mg of debrisoquin on two occasions. In healthy subjects, caffeine and debrisoquin metabolism and the oxidation and acetylation phenotypes were stable. In the patients with diabetes, the two tests showed a significant decrease in the glycosylated hemoglobin level and a significant increase in the 24-hour elimination rate of all caffeine metabolites. Most of the values were lower compared with those of healthy subjects during the first test. Because of these variations, caffeine cannot be used to determine the rapid or slow acetylator status in patients with diabetes. In contrast, neither the oxidation of debrisoquin nor the phenotypic expression was disturbed. These results reiterate the need for defining the administration conditions and surveying the drugs used in the treatment of diabetes mellitus complications.

A population and family study of N-acetyltransferase using caffeine urinary metabolites.

Population and family studies were undertaken to validate caffeine as a probe drug to establish the genetic status of rapid acetylators and slow acetylators. The acetylator status was established from the urinary metabolic ratio of 5-acetylamino-6-formylamino-3-methyluracil to 1-methylxanthine (AFMU/1X) after oral administration of caffeine. We confirmed a bimodal distribution (chi 2(1) = 229.48; p << 10(-9)) of the AFMU/1X ratio in 245 unrelated subjects. A third distribution did not significantly improve the fit to the data (chi 2(1) = 0.04; p = 0.84). Complex segregation analysis of 76 nuclear families confirmed the monogenic inheritance of N-acetyltransferase, with incomplete dominance of the rapid allele over the slow one. We observed a slight shift between the mean activities of heterozygous and homozygous rapid acetylators (t = 2.89; p < 0.01). However, the 30 obligate heterozygotes belonging to the 76 families were evenly distributed among the rapid acetylators and never located in a hypothetic intermediary group between slow acetylators and rapid acetylators.

Ultrastructural immunocytochemical localization of myosin in cultured fibroblastic cells.

Nonmuscle myosin in the cytoplasm of cultured fibroblastic cells has been localized using light and electron microscopic immunocytochemistry. Antibodies to purified fibroblast myosin were produced in goat and rabbit and purified by affinity chromatography. Light microscopic immunofluorescence localization showed patterns similar to those previously published. Electron microscopic localization using the ethyldimethyl aminopropyl carbodiimide-glutaraldehyde-saponin (EGS) fixation-permeabilization procedure and the ferritin bridge localization method produced quantifiable localization in intracellular sites with well-preserved ultrastructural morphology. Myosin was found to be a major component of the cytosol. It was distributed diffusely with no preferential localization on membranous organelles. Myosin was found to be slightly concentrated on the surface of microfilament-containing structures, including the subplasmalemmal microfilament mat and stress fibers, occasionally with an interrupted periodicity. However, no myosin was found in surface ruffles or microvilli. Morphometric quantitation showed that the majority of the cell's myosin was in the cytosol. This location is compatible with myosin being a component of the microtrabecular lattice of the cytoplasmic ground substance. The concentration of myosin in association with microfilaments was only twice that of the cytosol. This interpretation must be somewhat tempered by the possibility that some myosin bound to tightly packed actin may be inaccessible. The significance of this distribution of myosin in cell function is discussed.

Changes in the metabolic profile of equine muscle from birth through 1 yr of age.

The purpose of this study was to investigate metabolic changes in equine muscle from birth to 1 yr of age. Duplicate biopsies from the middle portion of the gluteus medius were obtained from a depth of 2 cm beneath the superficial fascia at 1 day, 7 days, 1 mo, 3 mo, 6 mo, and 1 yr of age in 11 quarter horses and at 1 day, 3 mo, 6 mo, and 1 yr of age in 5 Standardbreds. Muscle enzyme activities determined were citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, phosphorylase, and lactate dehydrogenase. Percent fast-twitch, fast-twitch high oxidative, and slow-twitch oxidative fiber types were determined using succinate dehydrogenase and myosin adenosinetriphosphatase (pH 9.4) histochemical stains. Histochemically determined muscle fiber-type percents did not change dramatically with increasing age. However, lactate dehydrogenase activity increased threefold in quarter horses and twofold in Standardbreds, and phosphorylase activity increased sixfold in quarter horses and sevenfold in Standardbreds from 1 day to 6 mo of age. Citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities decreased during the first 3 mo of age in quarter horses.

Skeletal muscle hypertrophy in rats having growth hormone-secreting tumor.

Plantaris muscle hypertrophy resulting from surgical ablation of the synergistic gastrocnemius muscle was compared between nontumor- and GH3 tumor-bearing rat groups (n = 8-10). GH3 cells (10(6)) were subcutaneously injected into 150-g female Wistar-Furth rats to initiate the tumor. After 17 days, the tumor-bearing rats gained 5.7 g body wt/day compared with 2.0 for the nontumor-bearing rats. The left gastrocnemius muscle was surgically removed from both nontumor and tumor groups. The gastrocnemius was removed from the tumor group after an increased growth rate was achieved. Seven days after surgery, the animals were killed and plantaris muscles were removed. The wet weight of the left plantaris muscle increased 45.6 and 44.0% over the unoperated contralateral control (right side) in the nontumor and tumor groups, respectively. The right control plantaris muscle in the tumor group was 63% heavier than the right control plantaris from the nontumor group; however, the proportion of body weight for plantaris was similar between the two groups. The effect of gastrocnemius ablation and tumor treatment on plantaris weight was additive, and the percent increase over the unoperated contralateral control side was similar between the two groups. These data demonstrate that skeletal muscle hypertrophy occurs in adult animals in which growth has been stimulated by a growth hormone-secreting tumor and could suggest that the muscle growth response caused by the tumor is operating by a mechanism different than work-induced hypertrophy.

Modulation of IGF mRNA abundance during stretch-induced skeletal muscle hypertrophy and regression.

Increased load on a muscle (synergistic overload or stretch) results in muscle hypertrophy. The expression of insulin-like growth factor I (IGF-I) mRNA in rat skeletal muscle is increased during synergistic overload-induced hypertrophy. Although it has also been established that fasting animals lose muscle protein, it has been shown that compensatory muscle hypertrophy occurs in adult fasting rats that are undergoing a net loss of body weight. The purpose of this investigation was to determine whether a relationship exists between IGF-I mRNA levels and muscle growth and regression. This was accomplished by examining whether IGF-I mRNA levels were altered during muscle hypertrophy after stretch and regression and the effect of fasting on IGF-I mRNA levels during stretch-induced hypertrophy. Patagialis (PAT) muscle weights increased 13 and 44% at 2 and 11 days of stretch, respectively. However, after removal of the stretch stimulus on day 11, PAT weights began to decrease, reaching control weights by 18 days. During the first time point (2 days), PAT muscle IGF-I mRNA remained constant. IGF-I mRNA abundance was threefold greater than contralateral control levels by 11 days of stretch. IGF-I mRNA levels decreased but remained significantly above control levels throughout the regression of hypertrophy (13, 18, and 25 days). Fasting did not alter PAT muscle response to stretch. After 11 days of stretch, PAT muscle weight increased 60% compared with contralateral control muscles and IGF-I mRNA levels increased three-fold. This study supports a role for IGF-I in muscle hypertrophy but not muscle atrophy.

Pharmacokinetic of a very low molecular weight heparin in chronic renal failure.

The pharmacokinetic characteristics of a low molecular weight heparin (LMWH) (Cy 222; mean mw: 2500 daltons) are studied in 24 patients with 3 degrees of chronic renal failure (CRF) stage I (creatinine clearance between 50 and 30 ml/mn), stage 2 (creatine clearance between 30 and 10 ml/mn), stage 3 (creatinine clearance below 10 ml/mn). Patients with CRF have significantly higher values of anti Xa activity at 3 hours (p less than 0.05), 5 hours (p less than 0.05), and at 8 hours (p less than 0.03) after injection than controls, CMAX values, VDSS and AUC do not differ, whereas patients with the highest stage of CRF are characterised by the most important t1/2 a (p less than 0.001) and the smallest total body clearance (p less than 0.01). Consequences of these disturbances of pharmacokinetic characteristics have to be evaluated before adequate posology of heparin fragments could be determined in patients with CRF.

Is debrisoquine hydroxylation modified during acute viral hepatitis?

Drug metabolism in the liver may be decreased during liver diseases. However, the extent of impairment of specific isozymes of cytochrome P450 is largely unknown. We have studied the debrisoquine hydroxylation capacity of 17 patients with acute viral hepatitis and 106 unrelated healthy subjects. Debrisoquine metabolic ratio was increased in extensive metabolizers (EM) with acute viral hepatitis as compared with healthy EMs (median metabolic ratio: 1.20 vs 0.84, P < 0.05). However, there was no difference in phenotype prevalence between patients and controls. Our results suggest that acute viral hepatitis only has a marginal effect on the activity of CYP2D6 and that substrates of this enzyme may be given in normal therapeutic doses to this category of patients.

Modulation of IGF mRNA abundance during muscle denervation atrophy.

Changes in skeletal muscle activity cause dramatic alterations in muscle mass. Increased load on a muscle (synergistic overload) results in muscle hypertrophy. During hypertrophy, skeletal muscle concentrations of insulin-like growth factors (IGF-I and IGF-II) mRNAs increase. To clarify the role of IGFs in regulating muscle mass, this study examined whether IGF-I and -II mRNA levels were altered during decreased muscle activity (denervation). Gastrocnemius weights decreased 4.2%, 7.7%, 18.1%, 27.7%, 35.1%, 45.0%, and 60.3% at 2, 3, 5, 7, 10, 12, and 17 d following denervation, respectively. Muscle DNA content remained constant throughout the first 12 d after surgery, but increased above control levels at day 17. During the first week after surgery, gastrocnemius IGF-II mRNA remained constant. However, IGF-II mRNA abundance was 2.5-fold greater than controls by 10 d of denervation, 3-fold by 12 d, and 6.8-fold by 17 d. On the other hand, IGF-I mRNA levels were not affected by denervation. In conclusion, although increased muscle activity results in a change of IGF-I mRNA expression, decreased muscle activity has no effect on IGF-I mRNA expression. In contrast, IGF-II mRNA levels increase with long-term denervation as well as with increased muscle activity. This study suggests that muscle activity may not be the only factor affecting IGF-I and -II expression.

Is insulin-like growth factor gene expression modulated during cardiac hypertrophy?

The goal of this study was to examine the relationship between work-induced cardiac hypertrophy and insulin-like growth factor-I and -II (IGF-I and -II) mRNA expression in adult female Sprague-Dawley rats. Increased cardiac work was induced by coarctation, which involved placing a constricting silk ligature around the abdominal aorta to increase peripheral resistance. Cardiac hypertrophy was determined by measuring in vivo left ventricular protein synthesis rates. There was a rapid increase in left ventricular weight (LV) [both absolute and relative to body weight (mg tissue.100 g-1 body weight)] following the coarctation surgery. By the third day following coarctation, LV weight was increased approximately 20% and reached 24% by the 10th day as compared with controls. Protein synthesis rates increased dramatically, reached a peak level at 3 d (133%.d-1) compared with 29% Ks.d-1 in the sham operated group and then began to slowly decrease toward control rates. The fractional synthesis rates of total protein in the LV were unchanged 1 d post-surgery. IGF-I mRNA content in the LV decreased to approximately 38% below the control content at day 1. However, by 3 d post-surgery IGF-I mRNA content increased to 30-50% above controls, were 31% above control by day 7, and remained elevated thereafter. On the other hand, IGF-II mRNA content remained constant throughout the 10 d post-surgery. Work-necessitated increase in cardiac protein mass may be mediated, in part, by a local autocrine/paracrine production of IGF-I.

Serum free 5-methoxypsoralen fraction in health and psoriasis: relationship with human serum albumin concentration.

Human serum albumin is known to be the main carrier of 5-methoxypsoralen (5-MOP) in serum. As hypoalbuminaemia may occur in psoriasis with inflammatory syndrome, variability of the free 5-MOP fraction in serum can be expected. The free 5-MOP fraction was determined by equilibrium dialysis in serum samples obtained from 18 psoriatic patients and 18 control subjects. The median free 5-MOP fraction was not significantly different in the psoriatic group (fu = 4.75%) than in the control group (fu = 5%). However, there was a significantly larger variability of the free fraction in the psoriatic group (2.7 to 8.6%) than in the healthy group (3.2 to 6.8%) (p = 0.002). The binding index of 5-MOP (ratio of bound to free concentrations) was correlated with human serum albumin level (r = 0.784). This work confirms that the 5-MOP fraction in human serum is principally serum albumin dependent, as has been described with in vitro models. Free drug monitoring of 5-MOP is discussed.