Partial chemical and functional characterization of milk whey products obtained by different processes

Whey protein samples (S-1 to S-5) were tested in vivo and in vitro for nutritional properties and selected bioactivities. Weanling male Wistar rats fed modified AIN-93G (12 g protein.100 g-1) diets for 21 days were used the in vivo studies. The nutritional parameters did not differ among the protein diets tested. Erythrocyte glutathione content was considered high and was higher for S-3, but liver glutathione was the same for all dietary groups. For S-3, cytokine secretion (IL-10 and TNF-α) by human peripheral blood mononuclear cells (in RPMI-1640 medium) was higher in the absence of antigen than in the presence of BCG antigen. Interleukin-4 secretion was repressed in all treatments. The IC50, whey protein concentration required to inhibit 50% of the melanoma cell proliferation, was 2.68 mg.mL-1 of culture medium for the S-3 sample and 3.66 mg.mL-1 for the S-2 sample. Based on these results, it was concluded that S-3 (whey protein concentrate enriched with TGF-β and lactoferrin) produced better nutritional and immunological responses than the other products tested.

Density and rheological parameters of goat milk

The rheological behavior and density of goat milk was studied as a function of solids concentration (10.5 to 50.0%) and temperature (273 to 331 k). Newtonian behavior was observed for values of total solids (TS) between 10.5 and 22.0% and temperatures from 276 to 331 k changing to pseudoplastic behavior without yield stress for TS from 25.0 to 39.4% at the same range of temperature. Goat milk with TS between 44.3 to 50.0% and temperatures of 273 to 296 k showed yield stress in addition to pseudoplastic behavior. At 303 to 331 k the power law model was observed again, without yield stress. The density of goat milk ranged from 991.7 to 1232.4 kg.m-3.

Effect of the addition of wheat fiber and partial pork back fat on the chemical composition, texture and sensory property of low-fat bologna sausage containing inulin and oat fiber

The objective of this work was to study the effect of adding wheat fiber and partial pork back fat on the quality characteristics of bologna sausage. The compound central rotating design was used with treatments containing fixed levels of inulin (5%) and oat fiber (1%) and variable levels of wheat fiber (0-4%) and pork back fat (0-10%). The pH and protein were similar in all the treatments, the fat was lower than the control treatment and the moisture content was higher than the control treatment (CF) without fibers. The wheat fiber increased the hardness and reduced cohesiveness and scores were given for overall impression. We found that it was possible to prepare low-fat bologna sausage with the addition of 6.58% fiber (5% inulin, 1% oat fiber and 0.58% wheat fiber), whilst retaining good sensory acceptability, thus reducing the pork back fat levels by between 25 and 42.75%.

Effect of previous chilling storage on quality loss in frozen (–20 °C) sierra (Scomberomorus sierra) muscle packed with a low-density polyethylene film containing butylated hydroxytoluene

Rancidity development during frozen storage (–20 °C) of sierra fish (Scomberomorus sierra) was studied. Fillets were packed in low-density polyethylene films with and without butylated hydroxytoluene added (BHT-LDPE and LDPE respectively). Fillets stored with no package were used as control. Special attention was given to the effect of previous ice storage (0, 3, 6, 9 and 15 days) on the quality of the frozen fish. Physical (pH and texture) and chemical (peroxide value, PV and thiobarbituric acid index, TBA-i) analyses were carried out. Lipid oxidation increased with ice storage time in fish muscle without film packing, being greater than the film packed muscle (with and without antioxidant). An effect of previous ice storage time was observed on the frozen product (in all treatments). However, fish muscle with film packing containing antioxidant showed less lipid deterioration. Under the conditions applied in this study, the plastic films with antioxidant prevented the lipids oxidation during the cold handling of the sierra muscle.

Purification, partial characterization and antimicrobial activity of Lectin from Chenopodium Quinoa seeds

Abstract A novel lectin was isolated from the seeds of Chenopodium quinoa. To achieve this end, the crude extract from the quinoa was submitted to two purification steps, Sephadex G50 and Mono Q. The hemagglutinating activity showed that this lectin agglutinates human erythrocytes. Its activity is inhibited by glucose and mannose, and remained stable under a wide range of pH levels and temperatures. The quinoa lectin was found to be a heterodimeric lectin of approximately 60 kDa, consisting of two subunits of approximately 25 kDa and 35 kDa. This lectin had its antimicrobial activity tested against several bacteria strains and effectively inhibited three strains. These strains were all Gram-negative, making this lectin a promising antimicrobial tool.

Physicochemical characteristics and sensory acceptability of ready-to-eat sliced frozen roast beef with partial reduction of sodium chloride

Abstract Sodium chloride in meat products provides microbiological stability and desirable technological and sensory effects. Therefore, the reduction of this ingredient is a challenge for the meat industry. The objective of this study was to evaluate the physicochemical and sensory characteristics of ready-to-eat sliced frozen roast beef with partial replacement of sodium chloride by a commercial additive mostly composed of potassium chloride. The analyses performed were chemical composition, cooking yield and post defrosting loss, microbiological evaluation and sensory analysis. There was higher moisture content (p < 0.05) in the control treatment (without the presence of the replacement additive) and all treatments were not different (p ≥ 0.05) in the cooking yield and in post-defrosting loss. The results of microbiological analysis are according to Brazilian Legislation. The sensory evaluation showed no difference between the control treatment and the T1 treatment (with the reduction of 35% of NaCl), while the T2 treatment (with reduction of 70% of NaCl) had the lowest average values in all attributes. The study showed that the reduction of 35% NaCl for commercial additive, mostly composed of potassium chloride, in roast beef is feasible since no changes were observed in sensory and technological characteristics evaluated.

Partial characterization of genes from the embryonic axis of Melanoxylon brauna Schott. (Leguminosae-Caesalpinioideae) seeds

The objective of this study was to partially characterize some genes involved in the desiccation tolerance of the embryonic axis of Melanoxylon brauna seeds subjected, or not, to oven fast-drying. Seeds were initially dried rapidly in an oven at 40 ºC, 50 ºC, 60 ºC, 70 ºC, and 80 °C, for 24, 48 and 72 h and then subjected to germination tests and moisture content determination. Degenerate primers were designed for 19 genes. The CDNA was used as a template for PCR amplifications using the degenerate primers, and the PCR products obtained were purified, cloned and sequenced. The seeds showed a gradual reduction in percent germination with increasing temperature and drying time. Nucleotide sequences of the cloned fragments related to genes CAT1, SPS1, Abi5, Transk and PM25 were obtained. The similarity analysis with the sequences deposited in databases revealed similarities with genes CAT1, SPS1, Transk and PM25 from other plant species. The nucleotide sequences obtained from the respective genes will be used for designing specific primers for gene expression analyses during seed germination in order to understand the causes for loss of physiological quality of Melanoxylon brauna seeds.

The Literary relations between La Fontaine and the "Astrée" of Honoré d'Urfé : a dissertation presented to the faculty of the department of philosophy of the university of Pennsylvania in partial fulfilment of the requirements for the degree of doctor of philosophy / by Walther P. Fischer

Collection : Publications of the University of Pennsylvania ; n° 6

Gypsy moths (Lymantria dispar) in the Niagara Region : population density variation, introduction of an entomopathogenic fungus (Entomophaga maimaiga) and occurrence of nuclear polyhedrosis virus

The gypsy moth, Lymantria dispar, a major defoliator of broad leaf trees, was accidentally introduced into North America in 1869. Much interest has been generated regarding the potential of using natural pathogens for biological control of this insect. One of these pathogens, a highly specific fungus, Entomophaga maimaiga, was accredited with causing major epizootics in populations of gypsy moth across the north-eastern United States in 1989 and 1990 and is thought to be spreading northwards into Canada. This study examined gypsy moth population densities in the Niagara Region. The fungus, .E.. maimaiga, was artificially introduced into one site and the resulting mortality in host populations was noted over two years. The relationship between fungal mortality, host population density and occurrence of another pathogen, the nuclear polyhedrosis virus (NPV), was assessed. Gypsy moth population density was assessed by counting egg masses in 0.01 hectare (ha) study plots in six areas, namely Louth, Queenston, Niagara-on-the-Lake, Shorthills Provincial Park, Chippawa Creek and Willoughby Marsh. High variability in density was seen among sites. Willoughby Marsh and Chippawa Creek, the sites with the greatest variability, were selected for more intensive study. The pathogenicity of E. maimaiga was established in laboratory trials. Fungal-infected gypsy moth larvae were then released into experimental plots of varying host density in Willoughby Marsh in 1992. These larvae served as the inoculum to infect field larvae. Other larvae were injected with culture medium only and released into control plots also of varying host density. Later, field larvae were collected and assessed for the presence of .E.. maimaiga and NPV. A greater proportion of larvae were infected from experimental plots than from control plots indicating that the experimental augmentation had been successful. There was no relationship between host density and the proportion of infected larvae in either experimental or control plots. In 1992, 86% of larvae were positive for NPV. Presence and intensity of NPV infection was independent of fungal presence, plot type or interaction of these two factors. Sampling was carried out in the summer of 1993, the year after the introduction, to evaluate the persistence of the pathogen in the environment. Almost 50% of all larvae were infected with the fungus. There was no difference between control and experimental plots. Data collected from Willoughby Marsh indicated that there was no correlation between the proportion of larvae infected with the fungus and host population density in either experimental or control plots. About 10% of larvae collected from a nearby site, Chippawa Creek, were also positive for .E.. maimaiga suggesting that low levels of .E.. maimaiga probably occurred naturally in the area. In 1993, 9.6% of larvae were positive for NPV. Again, presence or absence of NPV infection was independent of fungal presence plot type or interaction of these two factors. In conclusion, gypsy moth population densities were highly variable between and within sites in the Niagara Region. The introduction of the pathogenic fungus, .E.. maimaiga, into Willoughby Marsh in 1992 was successful and the fungus was again evident in 1993. There was no evidence for existence of a relationship between fungal mortality and gypsy moth density or occurrence of NPV. The results from this study are discussed with respect to the use of .E.. maimaiga in gypsy moth management programs.

Isolation and partial characterization of host cell wall surface glycoproteins: their involvement in agglutination, attachment and appressorium formation by piptocephalis virginiana

Cell surface proteins obtained by alkaline extraction from isolated cell walls of Mortierella pusilla and M. candelabrum, host and nonhost, respectively, to the mycoparasite, Piptocephalis virginiana, were tested for their ability to agglutinate mycoparasite spores. The host cell wall protein extract had a high agglutinating activity (788 a.u. mg- t ) as compared with the nonhost extract (21 a.li. mg- t ). SDS-polyacrylamide gel electrophoresis of the cell wall proteins revealed four protein bands, a, b, c, and d (Mr 117, 100, 85 and 64 kd, respectively) at the host surface, but not at the nonhost surface, except for the faint band c. Deletion of proteins b or c from the host cell wall protein extract significantly reduced its agglutinating activity. Proteins band c, obtained as purified preparations by a series of procedures, were shown to be two glycoproteins. Carbohydrate analysis by gas chromatography demonstrated that glucose and Nacetylglucosamine were the major carbohydrate components of the glycoproteins. It was further shown that the agglutinating activity of the pure preparation containing both band c was 500-850 times that of the single glycoproteins, suggesting the involvement of both glycoproteins in agglutination. The results suggest that the glycoproteins band c are the two subunits of agglutinin present at the host cell surface. The two glycoproteins band c purified from the host cell wall protein extract were further examined after various treatments for their possible role in agglutination, attachment and appressorium formation by the mycoparasite. Results obtained by agglutination and attachment tests showed: (1) the two glycoprotein-s are not only an agglutinin responsible for the mycoparasite spore agglutination, but may also serve as a receptor for the specific recognition, attachment and appressorium formation by the mycoparasite; (2) treatment of the rnycoparasite spores with various sugars revealed that arabinose, glucose and N-acetylglucosamine inhibited the agglutination and attachment activity of the glycoproteins, however, the relative percentage of appressorium formation was not affected by the above sugars; (3) the two glycoproteins are relatively stable with respect to their agglutinin and receptor functions. The present results suggest that the agglutination and attachment may be mediated directly by certain sugars present at the host and mycoparasite cell surfaces while the appressorlum formation may be the response of complementary combinations of both sugar and protein, the two parts of the glycoproteins at the interacting surfaces of two fungi.

Isolation and partial characterization of a complementary protein from the mycoparasite, Piptocephalis virginiana, which specifically binds to two glycoproteins b and c of the host cell surface

Presence of surface glycoprotein in Piptocephalis virginiana that recognizes the host glycoproteins band c, reported earlier from our laboratory, was detected by immunofluorescence microscopy. Germinated spores of P. virginiana treated with Mortierella pusilla cell wall protein extract, primary antibodies prepared against glycoproteins band c and FITC-goat anti-rabbit IgG conjugate showed fluorescence. This indicated that on the surfaces of the biotrophic mycoparasite P. virginiana , there might be a complementary molecule which recognizes the glycoproteins band c from M. pusilla. Immunobinding analysis identified a glycoprotein of Mr 100 kDa from the mycoparasite which binds with the host glycoproteins band c, separately as well as collectively. Purification of this glycoprotein was achieved by (i) 60% ammonium sulfate precipitation, (ii) followed by heat treatment, and (iii) Sephadex G-IOO gel filtration. The glycoprotein was isolated by preparative polyacrylamide gel electrophoresis by cutting and elution. The purity of the protein ·was ascertained by SDS-PAGE and Western blot analysis. Positive reaction to periodic acid-Schiff reagent revealed the glycoprotein nature of this 100 kDa protein. Mannose was identified as a major sugar component of this glycoprotein by using a BoehringerMannheim Glycan Differentiation Kit. Electrophoretically purified glycoprotein was used to raIse polyclonal antibody in rabbit. The specificity of the antibody was determined by dot-immunobinding test and western-blot analysis. Immunofluorescence mIcroscopy revealed surface localization of the protein on the germ tube of Piptocephalis virginiana. Fluorescence was also observed at the surfaceJ of the germinated spores and hyphae of the host, M. pusilla after treatment with complementary protein from P. virginiana, primary antibody prepared against the complementary protein and FITC-goat anti-rabbit IgG conjugate.

Mating behaviours of the field cricket (Gryllus integer) at different levels of male density

The reproductive behaviour of the field cricket, Gryllus integer, was systematically observed in indoor arenas to determine the extent of female Choice and male-male competition at different sex ratios representing two male densities (12:6 and 6:6). The costs and benefits to males and females in those two densities were analyzed according to the theory of the evolution o£ leks. Observations were conducted during the dark hours when most calling occurred since hourly rates of courtship song and mating did not fluctuate significantly over a 24 h period. Female mating rates were not significantly different between densities, therefore males at high densities were not advantaged because of increased female tendencies to mate when social stimulation was increased. Mean rates of acoustical signalling (calling and courtin"g) did not differ significantly between densities. Mean rates of fighting by males at the high density were significantly greater than those of males at the low density. Mating benefits associated with callin~courting and fighting were measured. Mating rates did not vary with rates of calling at either density. Calling was not a prerequisite to mating. Courtship song preceded all matings. There was a significant power fit between male mating and courting rates, and male mating and fighting rates at the low, but not at the high, density. Density differences in the benefits associated with increased courting and fighting may relate, in part, to greater economic defensibility and monopoly of females due to reduced male competition at the low density. Dominant males may be preferentially chosen by females or better able to monopolize mating opportunities than subordinate males. Three criteria were used to determine whether dominant males were preferentially chosen by females. The number of matings by males who won fights (within 30 min of mating) was significantly greater than the number of matings by males who were defeated in such fights. Mating rates did not vary significantly with rates of winning at either density. There was a significant power fit between male mating rates and the percentage of fights a male won (irrespective of his fighting-frequency) at the low density. The mean duration a male guarded the female after mating did not vary significantly between densities. There was a significant linear relationship between the duration a spermatophore was retained and the duration a male guarded the female after mating. Courtship song apparently stimulated spermatophore removal. Male guarding involved inter-male aggression and reduced courtship attempts by other males. Males at the high density received no apparent reproductive benefits associated with increased social stimulation. Conclusive evidence for preferential choice of males by females, using the criteria examined here, is lacking. Males at the lower density had fewer competitors and could monopolize females more effectively.

Partial characterization of the cloned dihydrofolate reductase gene of Saccharomyces cerevisiae

The cloned dihydrofolate reductase gene of Saccharomyces cerevisiae (DFR 1) is expressed in Escherichia coli. Bacterial strain JF1754 transformed with plasmids containing DFR 1 is at least 5X more resistant to inhibition by the folate antagonist trimethoprim. Expression of yeast DFR 1 in E. coli suggests it is likely that the gene lacks intervening sequences. The 1.8 kbp DNA fragment encoding yeast dhfr activity probably has its own promotor, as the gene is expressed in both orientations in E. coli. Expression of the yeast dhfr gene cloned into M13 viral vectors allowed positive selection of DFR 1 - M13 bacterial transfectants in medium supplemented with trimethoprim. A series of nested deletions generated by nuclease Bal 31 digestion and by restriction endonuclease cleavage of plasmids containing DFR 1 physically mapped the gene to a 930 bp region between the Pst 1 and Sal 1 cut sites. This is consistent with the 21,000 molecular weight attributed to yeast dhfr in previous reports. From preliminary DNA sequence analysis of the dhfr DNA fragment the 3' terminus of DFR 1 was assigned to a position 27 nucleotides from the Eco Rl cut site on the Bam Hi - Eco Rl DNA segment. Several putative yeast transcription termination consensus sequences were identified 3' to the opal stop codon. DFR 1 is expressed in yeast and it confers resistance to the antifolate methotrexate when the gene is present in 2 - 10 copies per cell. Plasmid-dependent resistance to methotrexate is also observed in a rad 6 background although the effect is somewhat less than that conferred to wild-type or rad 18 cells. Integration of DFR 1 into the yeast genome showed an intermediate sensitivity to folate antagonists. This may suggest a gene dosage effect. No change in petite induction in these yeast strains was observed in transformed cells containing yeast dhfr plasmids. The sensitivity of rad 6 , rad 18 and wild-type cell populations to trimethoprim were unaffected by the presence of DFR 1 in transformants. Moreover, trimethoprim did not induce petites in any strain tested, which normally results if dhfr is inhibited by other antifolates such as methotrexate. This may suggest that the dhfr enzyme is not the only possible target of trimethoprim in yeast. rad 6 mutants showed a very low level of spontaneous petite formation. Methotrexate failed to induce respiratory deficient mutants in this strain which suggested that rad 6 might be an obligate grande. However, ethidium bromide induced petites to a level approximately 50% of that exhibited by wild-type and rad 18 strains.