The identification of abnormal glycoforms of serum transferrin in carbohydrate deficient glycoprotein syndrome type i by capillary zone electrophoresis

One of the biochemical characteristics of carbohydrate deficient glycoprotein syndromes is the presence of abnormal glycoforms in serum transferrin. Both glycoform heterogeneity and variable site occupancy may, in principle, lead to the generation of a range of glycoforms which contain different numbers of sialic acid residues, and therefore variable amounts of negative charge. Capillary zone electrophoresis was used to resolve the glycoforms of normal human serum transferrin and also of a set of glycoforms which were prepared by digesting the sugars on the intact glycoprotein with sialidase. The sugars on the intact glycoprotein were also modified by a series of exoglycosidase enzymes to produce a series of neutral glycoforms which were also analysed by capillary zone electrophoresis. The oligosaccharide population of human serum transferrin was analysed by a series of mixed exoglycosidase digests on the released glycan pool and quantified using a novel HPLC strategy. Transferrin was isolated from carbohydrate deficient glycoprotein syndromes type I serum and both the intact glycoforms and released sugars were resolved and quantified. The data presented here confirm the presence of a hexa-, penta- and tetra-sialoforms of human serum transferrin in both normal and carbohydrate deficient glycoprotein syndrome type I serum samples. Consistent with previous reports carbohydrate deficient glycoprotein syndrome type I transferrin also contained a di-sialoform, representing a glycoform in which one of the two N-glycosylation sites is unoccupied, and a non-glycosylated form where both remain unoccupied. This study demonstrates that capillary zone electrophoresis can be used to resolve quantitatively both sialylated and neutral complex type glycoforms, suggesting a rapid diagnostic test for the carbohydrate deficient glycoprotein syndromes group of diseases.Abbreviations CDGS Carbohydrate Deficient Glycoprotein Syndrome - CZE Capillary Zone Electrophoresis - hTf human transferrin - gu HPLC glucose units - EOF electroosmotic flow. Nomenclature: for describing oligosaccharide structures: A(1,2,3,4) indicates the number of antennae linked to the t trimannosyl core - G(0–4) indicates the number of terminal galactose residues in the structure - F core fucose - B bisecting N-acetyl glucosamine (GlcNAc) - S sialic acid - Gal galactose; M - Man mannose     

Patterns of expression of human "Ia-like" antigens during the terminal stages of B cell development.

The "Ia-like" antigens characteristically present on the membrane of B lymphocytes were shown to be absent in most terminally differentiated Ig-producing plasma cells. This was most evident in analyses of myeloma plasma cells that uniformly lacked the Ia-antigens. Also, in B lymphoid cell lines, the lymphoblasts were uniformly Ia-positive but the plasma cells were negative. In contrast, however, the majority of plasma cells produced after pokeweed mitogen stimulation remained positive. Plasma cells in tonsil and in the tissue of peripheral blood of patients with Wladenstr?m's macroglobulinemia were also sometimes Ia-positive although the majority were negative. Studies of membrane IgM and IgD indicated a similar loss in some instances. However, the SIg and Ia antigens were not invariably associated. These results lead to the conclusion that Ia-antigens are differentiation antigens for B cells and are usually lost by the end stage cells in this series.     

Genetic and QTL analyses of seed dormancy and preharvest sprouting resistance in the wheat germplasm CN10955

The inheritance and genetic linkage analysis for seed dormancy and preharvest sprouting (PHS) resistance were carried out in an F₈ recombinant inbred lines (RILs) derived from the cross between “CN19055” (white-grained, PHS-resistant) with locally adapted Australian cultivar “Annuello” (white-grained, PHS-susceptible). Seed dormancy was assessed as germination index (GI7) while assessment for preharvest sprouting resistance was based on whole head assay (sprouting index, SI) and visibly sprouted seeds (VI). Segregation analysis of the F₂, F₃ data from the glasshouse and the RIL population in 2004 and 2005 field data sets indicated that seed dormancy and PHS resistance in CN19055 is controlled by at least two genes. Heritabilities for GI7 and VI were high and moderate for SI. The most accurate method for assessing PHS resistance was achieved using VI and GI7 while SI exhibited large genotype by environment interaction. Two quantitative trait loci (QTLs) QPhs.dpivic.4A.1 and QPhs.dpivic.4A.2 were identified. On pooled data across four environments, the major QTL, QPhs.dpivic.4A.2, explained 45% of phenotypic variation for GI7, 43% for VI and 20% for SI, respectively. On the other hand, QPhs.dpivic.4A.1 which accounted for 31% of the phenotypic variation in GI7 in 2004 Horsham field trial, was not stable across environments. Physical mapping of two SSR markers, Xgwm937 and Xgwm894 linked to the major QTL for PHS resistance, using Chinese Spring deletions lines for chromosome 4AS and 4AL revealed that the markers were located in the deletion bins 4AL-12 and 4AL-13. The newly identified SSR markers (Xgwm937/Xgwm894) showed strong association with seed dormancy and PHS resistance in a range of wheat lines reputed to possess PHS resistance. The results suggest that Xgwm937/Xgwm894 could be used in marker-assisted selection (MAS) for incorporating preharvest sprouting resistance into elite wheat cultivars susceptible to PHS.     

Predators in natural fragments: foraging ecology of wolves in British Columbia's central and north coast archipelago

Aim Predator–prey dynamics in fragmented areas may be influenced by spatial features of the landscape. Although little is known about these processes, an increasingly fragmented planet underscores the urgency to predict its consequences. Accordingly, our aim was to examine foraging behaviour of an apex mammalian predator, the wolf (Canis lupus), in an archipelago environment. Location Mainland and adjacent archipelago of British Columbia, Canada; a largely pristine and naturally fragmented landscape with islands of variable size and isolation. Methods We sampled 30 mainland watersheds and 29 islands for wolf faeces in summers 2000 and 2001 and identified prey remains. We examined broad geographical patterns and detailed biogeographical variables (area and isolation metrics) as they relate to prey consumed. For island data, we used Akaike Information Criteria to guide generalized linear regression model selection to predict probability of black‐tailed deer (main prey; Odocoileus hemionus) in faeces. Results Black‐tailed deer was the most common item in occurrence per faeces (63%) and occurrence per item (53%) indices, representing about 63% of mammalian biomass. Wolves consumed more deer on islands near the mainland (65% occurrence per item) than on the mainland (39%) and outer islands (45%), where other ungulates (mainland only) and small mammals replaced deer. On islands, the probability of detecting deer was influenced primarily by island distance to mainland (not by area or inter‐landmass distance), suggesting limited recolonization by deer from source populations as a causal mechanism. Main conclusions Although sampling was limited in time, consistent patterns among islands suggest that population dynamics in isolated fragments are less stable and can result in depletion of prey. This may have important implications in understanding predator–prey communities in isolation, debate regarding wolf–deer systems and logging in temperate rain forests, and reserve design.     

Mutational analysis of 85 mucopolysaccharidosis type I families: frequency of known mutations, identification of 17 novel mutations and in vitro expression of missense mutations

The lysosomal storage disorder, mucopolysaccharidosis type I (MPS I), is caused by a deficiency of the enzyme alpha-L-iduronidase, which is involved in the breakdown of dermatan and heparan sulphates. There are three clinical phenotypes, ranging from the Hurler form characterised by skeletal abnormalities, hepatosplenomegaly and severe mental retardation, to the milder Scheie phenotype where there is aortic valve disease, corneal clouding, limited skeletal problems, but no mental retardation. In this study, 85 MPS I families (73 Hurler, 5 Hurler/Scheie, 7 Scheie) were screened for 9 known mutations (Q70X, A75T, 474-2a>g, L218P, A327P, W402X, P533R, R89Q, 678-7g>a). W402X was the most frequent mutation in our population (45.3%) and Q70X was the second most frequent (15.9%). In 30 families, either one or both of the mutations were not identified, which accounted for 25.9% of the total alleles. Therefore, all 14 exons of the alpha-L-iduronidase gene were screened in these patients and 23 different sequence changes were found, 17 of which were previously unknown. The novel sequence changes include 4 deletions (153delC, 628del5, 740delC, 747delG), 5 nonsense mutations (Q60X, Y167X, Q400X, R619X, R628X), 6 missense mutations (C205Y, G208V, H240R, A319V, P496R, S633L), a splice site mutation (IVS12+5g>a), and a rare polymorphism (A591T). The polymorphism and novel missense mutations were transiently expressed in COS-7 cells and all of them except the polymorphism showed complete loss of enzyme activity. In total, 165 of the 170 mutant alleles were identified in this study and despite the high frequency of W402X and Q70X, the identification of many novel mutations unique to individual families further highlights the genetic heterogeneity of MPS I.     

Effects of elastic bands on force and power characteristics during the back squat exercise

Athletes commonly use elastic bands as a training method to increase strength and performance. The purpose of this study was to investigate the effect of elastic bands on peak force (PF), peak power (PP), and peak rate of force development (RFD) during the back-squat exercise (BSE). Ten recreationally resistance-trained subjects (4 women, 6 men, mean age 21.3 +/- 1.5 years) were tested for their 1 repetition maximum (1RM) in the BSE (mean 117.6 +/- 48.2 kg) on a Smith machine. Testing was performed on 2 separate days, with 2 sets of 3 repetitions being performed for each condition. Testing was conducted at 60% and 85% of 1RM with and without using elastic bands. In addition, 2 elastic band loading conditions were tested (B1 and B2) at each of the 2 resistances. No bands (NB) represents where all of the resistance was acquired from free-weights. B1 represents where approximately 80% of the resistance was provided by free-weights, and approximately 20% was provided by bands. B2 represents where approximately 65% of the resistance was provided by free-weights, and approximately 35% was provided from bands. The subjects completed the BSE under each condition, whereas PF, PP, and RFD was recorded using a force platform. There was a significant (p < 0.05) increase in PF between NB-85 and B2-85 of 16%. Between B1-85 and B2-85, PF was increased significantly by 5% (p < 0.05). There was a significant (p < 0.05) increase in PP between NB-85 and B2-85 of 24%. No significant differences were observed in RFD during the 85% conditions or for any of the measured variables during the 60% conditions (p < 0.05). The results suggest that the use of elastic bands in conjunction with free weights can significantly increase PF and PP during the BSE over free-weight resistance alone under certain loading conditions. The greatest differences are observed during the higher loading conditions, with the B1-85 condition appearing to be optimal for athletic performance of the ones we tested. The strength training professional could use variable resistance training (VRT) to increase PF and PP more than the traditional BSE can. VRT could also be used to train these 2 performance characteristics together, which might be especially useful in season, when weight-room training volume can sometimes be limited.     

A proposed method for determining peak power in the jump squat exercise

In recent years a great deal of research has been published using peak power (PP) in the jump squat (JS) exercise as a measure of athletic performance. However, no standardized method for the determination of PP exists at this time to accurately evaluate this variable. Our proposed method (PM) for determining PP (PPPM) in the JS uses the product of vertical ground reaction forces and velocity of the center of mass of both the subject and the external resistance of a loaded Olympic bar. Fifteen male subjects with a mean age of 27 +/- 3 years, weight of 78 +/- 17 kg, and height of 175 +/- 10 cm participated in this study. PP was measured in the JS at five different testing loads (30%, 35%, 40%, 45%, and 50% body weight) based on methods commonly discussed in the literature to compare PP results of previous methods to those obtained using the PM. Paired t-tests at different load levels were used for statistical analysis with an overall alpha = 0.05. The average PP among five testing loads, measured by the PM, was 3782 +/- 906 W. PP derived from the product of force and velocity of the bar alone was 72% lower than PPPM at 1057 +/- 243 W (P < 0.0001). The PP estimated by the product of bar velocity and vertical ground reaction forces of the bar plus the subject was 8% higher than PPPM at 4100 +/- 844 W (P = 0.0001). Our results indicate that using the methods traditionally reported in the literature may cause an overestimation of PP during athletic performance. Using the PM in future research will facilitate test validity and enable the generalization of results outside the scope of specific research projects.     

Static stretching impairs sprint performance in collegiate track and field athletes

Previous research has shown that static stretching (SS) can diminish the peak force output of stretch-shortening cycle actions while performing a dynamic warm-up (DW) protocol has been shown to enhance performance in similar activities. The purpose of this study was to establish whether the deleterious effects of SS would wash out the performance enhancements obtained from the DW. Eleven males and 11 females, who were athletes of a NCAA Division I track team, performed a DW followed with either a SS or rest (NS) condition. After warm-up was completed, three 40 m sprints were performed to investigate the effects of the SS condition on sprint performance when preceded by DW. Time(s) were obtained from timing gates placed at 0, 20, and 40 m respectively. Testing was conducted over 2 days with a 1 week washout period. Testing order was balanced to eliminate possible order effect. Time for the NS versus the SS group was significantly faster for the second 20 m with a time of 2.41 versus 2.38 seconds (P < or = .05), and for the entire 40 m with a time of 5.6 +/- 0.4 versus 5.7 +/- 0.4 seconds (P < or = .05). The results of this study suggest that performing a SS protocol following a DW will inhibit sprint performance in collegiate athletes.