Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Canonical correlation analysis of marine macrobenthos survey data

Canonical correlation analysis is applied to measurements of environmental variables and species distributions made during a survey of macrobenthos around a sewage-treatment farm drain. The implications of data reduction, necessary to enable the method to proceed, are discussed. The amount of data was reduced by discarding the rarest species, discarding species occurring at fewest stations, and including only those species and environmental variables which correlated highly with the greatest number of other variables. Only the third data-reduction scheme gave ecologically sensible results. Use of station scores on the first two canonical variates (CV1 and CV2) enabled the sampling grid to be divided into a group of nearshore stations, a group of intermediate depth, and a group of deep offshore stations. Loadings of environmental variables on the canonical variates were found to be unstable but correlations between these variables and canonical variates enabled the variates to be interpreted: CV1 as a gradient of depth and associated changes in sediment characteristics, CV2 with depth- and nutrient-related components, and CV3 as patchiness in sediment characteristics different from that normally expected with depth. Use of correlations between species and canonical variates enables definition of two major species groups, one confined to nearshore environments and a second offshore. These groups (and their sub-groups) related well to groups defined previously by hierarchical classification. It is concluded that, with careful attention to the method of data reduction, canonical correlation analysis can be an effective tool in the analysis of marine benthic survey data.     

Use of reversed phase HP liquid chromatography to assay conversion of N-acylglycines to primary fatty acid amides by peptidylglycine-alpha-amidating monooxygenase

Primary fatty acid amides (R-CO-NH2) and N-acylglycines (R-CO-NH-CH2-COOH) are classes of compounds that have only recently been isolated and characterized from biological sources. Key questions remain regarding how these lipid amides are produced and degraded in biological systems. Relative to the fatty acids, little has been done to develop methods to separate and quantify the fatty acid amides and N-acylglycines. We describe reversed phase HPLC methods for the separation of C2-C12 primary fatty acid amides and N-acylglycines and also C12-C22 fatty acid amides. Separation within each class occurs primarily on the basis of simple interactions between the acyl chain and the chromatographic stationary phase, but the polar headgroups on these and related fatty acids and N-acylethanolamides modulate the absolute retention in reversed phase mode. We use these methods to measure the enzyme-mediated, two-step conversion of N-octanoylglycine to octanoamide.     

Genetic and morphologic differentiation of Bolbophorus confusus and B. levantinus (Digenea: Diplostomatidae), based on rDNA SSU polymorphism and SEM

Metacercariae of Bolbophorus species are serious pathogens of farmed fish. Molecular diagnostic tools, capable of identifying and differentiating these parasites, may assist in the development of rationale control strategies. The rDNA 18S (small sub-unit: SSU) genes of adult B. confusus and B. levantinus obtained from a pelican, Pelecanus onocrotalus, and a night heron, Nycticorax nycticorax, respectively, were amplified, sequenced, and aligned. Based on this alignment, we developed a genetic differentiation assay between B. confusus and B. levantinus. These 2 species were compared genetically with the North American species B. damnificus and Bolbophorus sp. ('Type 2'). The relationship between species is outlined and discussed. In addition to the molecular study, specimens of B. confusus and B. levantinus were compared morphologically, using scanning electron microscopy. Morphologic analysis revealed interspecific differences in details of the holdfast organ and the position of the acetabulum.     

Alternative methods for disposal of spent laying hens: Evaluation of the efficacy of grinding,mechanical deboning,and of keratinase in the rendering process

Besides the challenges of mortality and litter disposal, the poultry industry must find economical means of disposing of laying hens that have outlived their productive lives. Because spent hens have low market value and disposing of them by composting and burial is often infeasible, finding alternative disposal methods that are environmentally secure is prudent. The feasibility of grinding or mechanically deboning spent hens with and without prior mechanical picking was evaluated for the production of various proteinaceous by-product meals. The end products were analyzed for nutrient content and found to be high in protein (35.3–91.9% CP) and, with the exception of the feathers, high in fat (24.1–58.3%), making them potentially valuable protein and energy sources. After considering physical and economic feasibility, mechanical deboning was determined to be a logical first step for the conversion of spent hens into value-added by-product meals. Because the hard tissue fraction (primarily feathers, bones, and connective tissue) generated by mechanically deboning the hens presents the greatest challenge to their utilization as feedstuffs, attention was focused on technologies that could potentially improve the nutritional value of the hard tissue for use as a ruminant protein source. Traditional hydrolysis of this hard tissue fraction improved its pepsin digestibility from 74% to 85%; however, subsequent keratinase enzyme treatment for 1 h, 2 h, 4 h, or 20 h after steam hydrolysis failed to improve the pepsin or amino acid digestibility any further (P > 0.10). Enzyme hydrolysis did, however, increase the quantities of the more soluble protein fractions (A: 45.5, 46.6, 52.8, 51.6, and 55.8% of CP; B₁: 3.2, 9.8, 6.0, 4.6, and 4.1% of CP; B₂: 11.7, 18.1, 22.8, 29.6, and 22.0% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively) and reduced quantities of the less soluble fractions (B₃: 30.2, 18.1, 10.8, 5.5, and 10.2% of CP; C: 9.4, 7.5, 7.6, 8.8, and 7.9% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively). The protein digestibility of the steam hydrolyzed hard tissue fraction from the mechanical deboning of spent hens was found to be comparable to the digestibility of feather meal, but post-hydrolysis keratinase treatment did not improve feeding value for ruminants.     

Nutritional challenges during development induce sex-specific changes in glucose homeostasis in the adult sheep

The early-life environment has implications for risk of adult-onset diseases, such as glucose intolerance, insulin insensitivity, and obesity, effects that may occur with or without reduced birth weight. We determined the consequences of nutrient restriction in early gestation and early postnatal life and their interactions on postnatal growth, body composition, and glucose handling. Ewes received 100% (C, n = 39) or 50% nutritional requirements (U, n = 41) from 1 to 31 days gestation and 100% thereafter. Male and female offspring (singleton/twin) from C and U ewes were then fed either ad libitum (CC n = 22, UC n = 19) or to reduce body weight to 85% of target from 12 to 25 wk of age (CU n = 17, UU n = 22) and ad libitum thereafter. At 1.5 and 2.5 yr, glucose handling was determined by area under the curve (AUC) for glucose and insulin concentrations following intravenous glucose (0.5 g/kg body wt). Insulin sensitivity was determined at 2.5 yr following intravenous insulin (0.5 IU/kg). In females, postnatal undernutrition reduced (P < 0.05) glucose AUC at both ages, regardless of prenatal nutrition. Postnatal undernutrition did not affect insulin secretion in females but enhanced insulin-induced glucose disappearance in singletons. Poor early postnatal growth was associated with increased fat in females. In males, glucose tolerance was unaffected by undernutrition despite changes in insulin AUC dependent on age, treatment, and single/twin birth. Nutrition in early postnatal life has long-lasting, sex-specific effects on glucose handling in sheep, likely due, in females, to enhanced insulin sensitivity. Improved glucose utilization may aid weight recovery but have negative implications for glucose homeostasis and body composition over the longer term.