Effects of Oxidation on Water Distribution and Physicochemical Properties of Porcine Myofibrillar Protein Gel

The effects of hydroxyl radicals induced oxidation on water distribution of porcine myofibrillar protein (MP) gels were investigated using nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI), and relationship between MP gel properties and water distribution was analyzed with Pearson’s correlation. Results of NMR spin-spin relaxation time (T₂) and MRI suggested that, comparing with unoxidized MP gel, the population of immobile water of gel (P₂₂) decreased by 18.81 %, while that of free water (P₂₃) increased by 85.56 %, and grey scale value decreased by 18.52 %, after being oxidized with 20 mM H₂O₂, respectively. Images of scanning electron microscope (SEM) showed the highest degree of oxidation led to the most coarse and porous gel-network. Gel properties including WHC, whiteness and strength of MP gels decreased by 14.65 %, 2.83 % and 52.74 % after being oxidized with 20 mM H₂O₂ (p?<?0.05), respectively. Pearson’s correlation analysis showed oxidation, gel-properties and NMR results were highly correlated (p?<?0.05). Therefore, we hypothesized that hydroxyl radicals induced oxidation resulted in immobile water shifting to free water partly, thus reduced the abundance of water and gel properties of gels.     

Low genic variation in male-reproductive-tract proteins of Drosophila melanogaster and D. simulans

We report results, using two-dimensional gel electrophoresis (2DE), of natural population surveys of allelic variation in approximately 300 male-reproductive-tract polypeptides in both Drosophila melanogaster and its sibling species, D. simulans. Despite our efforts to maximize operational sensitivity of our 2DE gels to polymorphism, variation estimates in both species were low (proportion of polymorphic loci [P] = 9%, and average heterozygosity [H] = 1%-3%), compared with those by one-dimensional gel electrophoresis (1DE) (P = 29%-55%; H = 8%-19%) in the same populations. However, H of polymorphic loci was very similar for 2DE and 1DE proteins; and for 17 of a total of 54 polymorphic proteins, 2DE detected three or four distinct alleles. The results suggest that the differing levels of variability widely seen with 1DE and 2DE are real and reflect differing intensities of functional constraint between different classes of structural loci. However, the alternative possibility remains that 2DE has a greater between-locus unevenness of variant detection sensitivity than does 1DE.     

1H NMR studies: dynamics of water in gelatin

 Proton magnetic resonance was used to characterize the dynamics of water in gelatin. Both sol and gel states were investigated. Transverse relaxation rates (R ₂) were dependent on the proton frequency measurement. (R ₂) measured with the Carr-Purcell-Meiboom-Gill pulse sequence was dependent on pulse spacing. These observations were interpreted in terms of chemical exchanges between water protons and those of the macromolecules in the sol state, whereas in the gel state the contribution of diffusion through microheterogeneities in the sample seems to provide an additional transverse relaxation mechanism. Received: 10 May 1999 / Revised version: 13 December 1999 / Accepted: 25 January 2000     

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresis-nitrocellulose blotting

An optimized multilocus enzyme electrophoresis method, which involves polyacrylamide-agarose gel electrophoresis followed by electrophoretic transfers on nitrocellulose sheets, was developed for the analysis of enzyme polymorphism in several aerobic and anaerobic bacterial species including Staphylococcus aureus, Streptococcus pneumoniae, S. agalactiae, Klebsiella pneumoniae and K. oxytoca, Clostridium bifermentans and C. sordellii, and Prevotella bivia. Serial electrophoretic transfers (during 5-15 min each) from a single polyacrylamide gel could be achieved for most enzymes studied, and allowed an increased definition of enzyme bands on nitrocellulose as compared to migration gels. Four enzymes, which could not be blotted in such conditions, could still be stained in gels after blotting. Thus, the method allowed the combined analysis of several enzymes after a single gel electrophoresis separation. The analysis of enzyme polymorphism in the various species studied raised the interest of polymorphic loci such as esterase or glutamic-oxaloacetic transaminase for epidemiologic studies. The method characterized a genetic diversity of enzyme loci of S. pneumoniae higher than previously reported, and is thus convenient for the analysis of genetic relationships between related isolates. Since the present method reduces the tediousness of multilocus enzyme electrophoresis and requires experimental conditions that are not specific for the bacterial population studied, it may be proposed for rapid population genetics analysis of a wide variety of bacteria.     

Concordance between life history traits, invasion history, and allozyme diversity of the Everglades invader Melaleuca quinquenervia

During the century following its initial introduction in 1886, the Australian tree Melaleuca quinquenervia (Myrtaceae) dispersed from a few introduction points to occupy over 200,000 ha, primarily in historic Everglades wetlands of southern Florida. Cellulose acetate gel electrophoresis (CAGE) was used to investigate the allozyme diversity and population genetic structure of 208 individuals in a dozen populations resulting from this invasion. The analyses showed that these populations have a high (82%) rate of polymorphic loci and an average of 2 alleles/locus. There was substantial heterozygosity (mean H_e = 0.356), which concords well with recent studies reporting a greater number of introduction events and sources than generally recognized. The introduction history and distributional patterns within Florida have led to geographic structuring (G_ST = 0.419) in which the Gulf Coast metapopulation has a greater effective number of alleles and greater heterozygosity than the Atlantic Coast metapopulation. The gene diversity in M. quinquenervia was comparable to other tropical woody species. Its strong population divergence was reminiscent of pioneer species and consistent with its status as a plant invader in Florida.     

SYNERESIS AND SWELLING OF GELATIN

1. When solid blocks of isoelectric gelatin are placed in cold distilled water or dilute buffer of pH 4.7, only those of a gelatin content of more than 10 per cent swell, while those of a lower gelatin content not only do not swell but actually lose water. 2. The final quantity of water lost by blocks of dilute gelatin is the same whether the block is immersed in a large volume of water or whether syneresis has been initiated in the gel through mechanical forces such as shaking, pressure, etc., even in the absence of any outside liquid, thus showing that syneresis is identical with the process of negative swelling of dilute gels when placed in cold water, and may be used as a convenient term for it. 3. Acid- or alkali-containing gels give rise to greater syneresis than isoelectric gels, after the acid or alkali has been removed by dialysis. 4. Salt-containing gels show greater syneresis than salt-free gels of the same pH, after the salt has been washed away. 5. The acid and alkali and also the salt effect on syneresis of gels disappears at a gelatin concentration above 8 per cent. 6. The striking similarity in the behavior of gels with respect to syneresis and of gelatin solutions with respect to viscosity suggests the probability that both are due to the same mechanism, namely the mechanism of hydration of the micellæ in gelatin by means of osmosis as brought about either by diffusible ions, as in the presence of acid or alkali, or by the soluble gelatin present in the micellæ. The greater the pressures that caused swelling of the micellæ while the gelatin was in the sol state, the greater is the loss of water from the gels when the pressures are removed. 7. A quantitative study of the loss of water by dilute gels of various gelatin content shows that the same laws which have been found by Northrop to hold for the swelling of gels of high concentrations apply also to the process of losing water by dilute gels, i.e. to the process of syneresis. The general behavior is well represented by the equations: See PDF for Equation and See PDF for Equation where P ₁ = osmotic pressure of the soluble gelatin in the gel, P ₂ = stress on the micellæ in the gelatin solution before setting, K_e = bulk modulus of elasticity, V_o = volume of water per gram of dry gelatin at setting and V_e = volume of water per gram of gelatin at equilibrium.     

Impacts of predation on dynamics of age-structured prey: Allee effects and multi-stability

With a series of mathematical models, we explore impacts of predation on a prey population structured into two age classes, juveniles and adults, assuming generalist, age-specific predators. Predation on any age class is either absent, or represented by types II or III functional responses, in various combinations. We look for Allee effects or more generally for multiple stable steady states in the prey population. One of our key findings is the occurrence of a predator pit (low-density ??refuge?? state of prey induced by predation; the chance of escaping predation thus increases both below and above an intermediate prey density) when only one age class is consumed and predators use a type II functional response ??this scenario is known to occur for an unstructured prey consumed via a type III functional response and can never occur for an unstructured prey consumed via a type II one. In the case where both age classes are consumed by type II generalist predators, an Allee effect occurs frequently, but some parameters give also rise to a predator pit and even three stable equilibria (one extinction equilibrium and two positive ones??Allee effect and predator pit combined). Multiple positive stable equilibria are common if one age class is consumed via a type II functional response and the other via a type III functional response??here, in addition to the behaviours mentioned above one may even observe three stable positive equilibria????double?? predator pit. Some of these results are discussed from the perspective of population management.     

Steady-state and pulsed NMR studies of gelation in aqueous agarose

Steady-state and pulsed NMR techniques have been used to investigate molecular motion in sols and gels of agarose. In passing through the sol–gel transition, the molecular mobility of water molecules is reduced only by a small amount, whereas motion of the polymer chains is greatly attenuated. The results are discused in terms of the network theory of gelation, with references to the role of water in the process and the nature of the “junction zones” between polymer chains. T₂ and line-width measurements are dominated by exchange broadening. The effects of exchange rate and differences in relaxation time between the exchanging sites are discussed. The temperature hysteresis behavior of agarose gels has been investigated and the effects of “ageing” correlated with changes in nuclear relaxation times. The synergistic increase in gel strength obtained on adding locust bean gum (LBG) to agarose has been investigated. The results indicate that LBG does not form double-helix junctions and may decrease rates of gelation by steric effects. At high agarose concentration, the LBG remains mainly in solution in interstitial water, but at low agarose concentration, it is suggested that the LBG can link gel aggregates together into a self-supporting structure, producing a synergistic increase in gel strength. Comparisons have been made between the nature of the agarose–LBG interaction and agarose–cellulose interactions in biological systems.     

Further studies on bovine serum amylase. 2. Affinity electrophoresis

The polymorphism of bovine serum amylase, which is controlled by the Ami locus, has previously only been demonstrated by starch gel electrophoresis. The addition of maltose to starch gels has been demonstrated to inhibit any subsequent separation of the Ami isozymes by starch gel electrophoresis. When electrophoresis was conducted in a support medium in the absence of starch no polymorphic variation was detected amongst samples from animals of different Ami phenotypes. The addition of starch to agarose gels has been shown to facilitate the subsequent detection of the Ami polymorphism by agarose/starch gel electrophoresis. The electrophoretic resolution of the Ami isozymes has been demonstrated to depend upon differences in affinity for starch rather than differences in net charge. The starch gel electrophoretic separation of the Ami isozymes is. therefore, another example of affinity electrophoresis. All the Ami amylases have been shown to share a common isoelectric point of pH 3.5.     

Controlled Release of Food Lipids Using Monoglyceride Gel Phases Regulates Lipid and Insulin Metabolism in Humans

We describe a solid vegetable oil–water gel structure which is stabilized through the use of low concentrations of monoglycerides, containing no added trans fats or saturated fats. The resulting structure consists of oil droplets encapsulated in self-assembled crystalline monoglyceride multilayers, surrounded by a continuous water phase. Acute ingestion human feeding trials indicated that the serum triglyceride loading was significantly lower after ingestion of the structured gel rather than a simple oil–water mixture, resulting in an attenuated increase in serum insulin. This indicates the effectiveness of encapsulation in modulating blood lipid and insulin response in humans, and suggests a strategy that can be employed for the controlled release of food macronutrients. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Consequences of the Allee Effect and Intraspecific Competition on Population Persistence under Adverse Environmental Conditions

The impact of intraspecific interactions on ecological stability and population persistence in terms of steady state(s) existence is considered theoretically based on a general competition model. We compare persistence of a structured population consisting of a few interacting (competitive) subpopulations, or groups, to persistence of the corresponding unstructured population. For a general case, we show that if the intra-group competition is stronger than the inter-group competition, then the structured population is less prone to extinction, i.e. it can persist in a parameter range where the unstructured population goes extinct. For a more specific case of a population with hierarchical competition, we show that relative viability of structured and unstructured populations depend on the type of density dependence in the population growth. Namely, while in the case of logistic growth, structured and unstructured populations exhibit equivalent persistence; in the case of Allee dynamics, the persistence of a hierarchically structured population is shown to be higher. We then apply these results to the case of behaviourally structured populations and demonstrate that an extreme form of individual aggression can be beneficial at the population level and enhance population persistence.     

Effects of sucrose and urea on soy hull pectic polysaccharide gel induced by d-glucono-1,5-lactone

Gelation properties of pectic polysaccharide extracted with ammonium oxalate from soybean hulls assisted by microwave were seldom studied. Water mobility in soy hull pectic polysaccharide (SHPP) was firstly studied by low field NMR. d-Glucono-1,5-lactone (GDL) and sucrose both could decrease spin–spin relaxation times (T₂) of SHPP solutions which indicated the SHPP network formed. Rheological analysis conformed that SHPP gel was formed induced by GDL and enhanced by sucrose. Urea can increase T₂ and collapse the network of SHPP. TGA was used to draw the profiles of water desorption from SHPP solutions or gels, during heating at a controlled rate. It was found that sucrose increased the bound water content and urea acted a conversely role. Hydrogen bond is the main force to maintain SHPP gel network.     

A comparison of AFLPs and microsatellites to identify the population structure of brown trout (Salmo trutta L.) populations from Hardangervidda, Norway

A total of 495 fish from 11 Hardangervidda lakes were genotyped in order to compare amplified fragment length polymorphisms (AFLP) and microsatellites in terms of their capacity to infer population genetic structure. The 11 microsatellites used in this study gave a greater polymorphism information content and greater gene diversity, with an average of 14.8 alleles per locus, than the six AFLP primer combinations used. However, the AFLPs resulted in 178 polymorphic loci and a 3.1 times larger marker index (effective multiplex ratio multiplied with the gene diversity). Comparable population structuring, for example in terms of distinguishing fish from the different river systems, was obtained with both marker systems. An AFLP and microsatellite multilocus Bayesian assignment test with the structure program divided the fish into six groups largely concurrent with main branches on a population neighbour-joining tree. Yet, the admixture status of individuals is mostly contradictory in the AFLP and the microsatellite analyses. The results are discussed concerning migration between lake populations.     

Rheologic behavior of deoxyhemoglobin S gels

The physical properties of deoxyhemoglobin S gels formed from solutions at concentrations and temperatures approaching those in vivo have been characterized by stress relaxation using a rotational rheometer. Gels were annealed in the rheometer and then subjected to a constant shear strain; thereafter the stress sustained was followed with time. Gels with solid-like behavior held stress indefinitely, and were characterized by yield temperature (the temperature at which stress decreased). Gels with less solid behavior were unable to hold target stress, and were characterized by yield stress (maximum stress attained) and equilibrium stress (final stress held). The samples were ultracentrifuged to calculate pellet and polymer masses. The solidity of the gels, as measured by yield temperature or yield stress, was related to the initial hemoglobin concentration, pellet and polymer masses, shear history, temperature, and the temperature and time of annealing. Solidity increased significantly with time when gels were annealed at 37 degrees C, whereas, when annealed at 25 degrees C, no or minimal increases in solidity were noted. Studies suggest that polymerization occurs rapidly and is completed early in or before the gel annealing period and that the increase in solidity with time of annealing is mainly due to factors other than polymer mass, i.e. alignment, increasing bond strength, water loss. The chemical activity of deoxyhemoglobin S did not affect the solidity of the formed gels. When the resultant polymer masses were comparable, gels formed from samples with albumin present (higher initial total protein concentration, but lower initial deoxyhemoglobin S concentration), had the same behavior as gels formed from solutions with higher initial hemoglobin S concentration. These findings demonstrate that gel annealing conditions must be standardized when comparing the rheologic behaviors of deoxyhemoglobin S gels and indicate that the gel's physical properties (influenced by polymer mass, shear history, annealing time) must be considered in understanding pathophysiology of sickling disorders.     

Genome‐wide population structure and admixture analysis reveals weak differentiation among Ugandan goat breeds

Uganda has a large population of goats, predominantly from indigenous breeds reared in diverse production systems, whose existence is threatened by crossbreeding with exotic Boer goats. Knowledge about the genetic characteristics and relationships among these Ugandan goat breeds and the potential admixture with Boer goats is still limited. Using a medium‐density single nucleotide polymorphism (SNP) panel, we assessed the genetic diversity, population structure and admixture in six goat breeds in Uganda: Boer, Karamojong, Kigezi, Mubende, Small East African and Sebei. All the animals had genotypes for about 46 105 SNPs after quality control. We found high proportions of polymorphic SNPs ranging from 0.885 (Kigezi) to 0.928 (Sebei). The overall mean observed (H_O) and expected (H_E) heterozygosity across breeds was 0.355 ± 0.147 and 0.384 ± 0.143 respectively. Principal components, genetic distances and admixture analyses revealed weak population sub‐structuring among the breeds. Principal components separated Kigezi and weakly Small East African from other indigenous goats. Sebei and Karamojong were tightly entangled together, whereas Mubende occupied a more central position with high admixture from all other local breeds. The Boer breed showed a unique cluster from the Ugandan indigenous goat breeds. The results reflect common ancestry but also some level of geographical differentiation. admixture and f₄ statistics revealed gene flow from Boer and varying levels of genetic admixture among the breeds. Generally, moderate to high levels of genetic variability were observed. Our findings provide useful insights into maintaining genetic diversity and designing appropriate breeding programs to exploit within‐breed diversity and heterozygote advantage in crossbreeding schemes.     

Modeling Black-Tailed Deer Population Dynamics Using Structured and Unstructured Approaches

Abstract: We used cohort analysis to reconstruct the female segment of a Columbian black-tailed deer (Odocoileus hemionus columbianus) population from 1979 to 2000 in the western Cascades of Washington, USA. We used reconstructed population estimates and age class representations to analyze relationships between population change and female density, forage availability, and weather influences. We applied stage structured and unstructured modeling approaches and used information-theoretic methods to select the best models. We used habitat covariates to develop predictive functions for fertility and survival parameters in structured models. The best structured and unstructured models were composed of combinations of factors including population density, forage availability, and winter weather. Structured and unstructured models could assist with management of black-tailed deer by providing the ability to predict deer population change given covariate values.     

Effect of physico-chemical properties of polyacrylamide gel media on the growth of Escherichia coli colonies]

We studied the growth of Escherichia coli LE-392 colonies on polyacrylamide gels (PAAG) depending on the physico-chemical properties of the latter, i.e. polymer concentration in the gel, swelling degree, bound water content (fm), spin-lattice relaxation and spin-spin relaxation times of water molecule protons, and modulus of elasticity (G0). S- or R-type colonies formed depending on gel properties; the diametral growth rate of S colonies was 3 times less compared with that on the control agar medium (Tryptose broth). The procedure is proposed for preparation of PAAG which rules out syneresis. Functional relations between the polymer concentrations in uniformly swelling gels and concentrations of copolymers in the reaction mixture, fm and G0 were revealed. The fm and G0 parameters can be used for controlling the quality of PAAG.     

Proton Nuclear Magnetic Resonance Relaxation Measurements in Frog Muscle

Proton nuclear magnetic resonance (NMR) relaxation measurements are reported for frog muscle as a function of temperature and Larmor frequency. Each T_1ρ, T₂, and T₁ measurement covered a time domain sufficient to identify the average relaxation time for most intracellular water. Using regression analysis the data were fit with a model where intracellular water molecules are exchanging between a large compartment in which mobility is similar to ordinary water and a small compartment in which motion is restricted. The regression results suggest that: the restricted compartment exhibits a distribution of motions skewed toward that of free water; the residence time of water molecules in the restricted compartment is approximately 1 ms; and, the activation entropy for some water molecules in the restricted compartment is negative.     

Out crosses between seasonally different segments of a Pacific salmon population reveal local adaptation

In salmon populations, local adaptation to seasonally varying incubation temperature is characterized by temperature-adjusted development times [measured in degree days – accumulated temperature units (ATUs)] that differ between control and F₁ hybrid crosses that were made between temporally separated population segments, a contrast not expected in a panmictic population. We examined adaptation of embryo development time to seasonally cooling temperature in a population of pink salmon by estimating genetic components of variation in control and hybrid F₁ crosses made between members of early- and late-spawning subpopulations, and replicated our observations in independent odd- and even-year brood lines. In each brood line, both sire and dam components of variation of development time were significant and accounted for a substantially larger portion of variation than their interactions, which suggested that natural selection has acted primarily on additive genetic variation. The implications of these results are that (1) spatially or temporally proximate salmon populations may be structured by distinct adaptations; (2) artificial relaxation of local geneflow barriers may lead to depression of fitness; and (3) populations of salmon genetically structured by local adaptation may carry variation that enhances their persistence during rapid climate change.     

Population structure and genetic diversity of metamorphic and paedomorphic populations of the tiger salamander,Ambystoma tigrinum

Genetic relationships, population subdivision and genetic diversity were estimated from mtDNA and allozyme data for two subspecies of tiger salamander, one of which is obligately metamorphic and the other polymorphic for paedomorphosis (larval reproduction). Far greater genetic differentiation exists between subspecies than within subspecies, suggesting that the subspecies have evolved in allopatry. Values of F_st calculated from both mtDNA and allozymes were greater than 0.400 for each subspecies. Significant population subdivision was detected even on a microgeographic scale. This extensive population subdivision indicates that populations can respond to extremely localized selection pressures. In the case of paedomorphosis, populations in permanent water should evolve paedomorphosis as long as the appropriate genes exist. For both mtDNA and allozymes, comparisons of population structure within the polymorphic subspecies and between polymorphic and metamorphic subspecies reveal no discernible effects of paedomorphosis. However, a comparison of paedomorphic and metamorphic populations of the polymorphic subspecies showed significantly higher mtDNA diversity in paedomorphic populations. The discrepancy between the allozyme and mtDNA results may be due to the lower effective population size of mtDNA compared to autosomal genes.