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Differential involvement of cell surface sialic acid residues in wheat germ agglutinin binding to parental and wheat germ agglutinin-resistant Chinese hamster ovary cells.

Two Chinese hamster ovary (CHO) cell mutants selected for resistance to wheat germ agglutinin (WGA) have been shown to exhibit defective sialylation of membrane glycoproteins and a membrane glycolipid, GM3. The mutants (termed WgaRII and WgaRIII) have been previously shown to belong to different genetic complementation groups and to exhibit different WGA-binding abilities. These mutants and a WGA-resistant CHO cell mutant termed WgaRI (which also possesses a surface sialylation defect arising from a deficient N-acetylglucosaminyltransferase activity), have enabled us to investigate the role of sialic acid in WGA binding at the cell surface. Scatchard plots of the binding of 125I-WGA (1 ng/ml to 1 mg/ml) to parental and WgaR CHO cells before and after a brief treatment with neuraminidase provide evidence for several different groups of sialic acid residues at the CHO cell surface which may be distinquished by their differential involvement in WGA binding to CHO cells.

Animals↗

Combining wheat bran with resistant starch has more beneficial effects on fecal indexes than does wheat bran alone.

BACKGROUND: Wheat bran (WB) increases fecal bulk and hastens colonic transit, whereas resistant starch (RS) has effects on colonic fermentation, including increasing concentrations of butyrate. OBJECTIVE: We hypothesized that a diet combining WB with RS would produce more favorable changes in fecal variables (eg, fecal bulk, rapid transit time, lower pH, and higher butyrate) than would WB alone. DESIGN: This was a randomized crossover block-design study for which 20 volunteers with a family history of colorectal cancer were recruited. The study included 3 diets: control, WB (12 g fiber/d), and WBRS (12 g WB fiber/d plus 22 g RS/d), each continued for 3 wk. In each diet, the major source of protein was lean red meat. During 5 consecutive days (days 15-19) of each dietary period, the subjects collected their total fecal output for analysis. RESULTS: The WB diet resulted in greater fecal output (by 23% and 21% for wet and dry weights, respectively) and a lesser transit time (-11 h) than did the control diet but did not have major effects on fermentation variables. Compared with the control diet, the WBRS diet resulted in greater fecal output (by 56%) and a shorter transit time (-10 h), lower fecal pH (-0.15 units), higher fecal concentration (by 14%) and daily excretion (by 101%) of acetate, higher fecal concentration (by 79%) and daily excretion (by 162%) of butyrate, a higher fecal ratio of butyrate to total short-chain fatty acids (by 45%), and lower concentrations of total phenols (-34%) and ammonia (-27%). CONCLUSIONS: Combining WB with RS had more benefits than did WB alone. This finding may have important implications for the dietary modulation of luminal contents, especially in the distal colon (the most common site of tumor formation).

Adult↗

Bioconversion of wheat straw and wheat straw components into single-cell protein.

Several fungi (Aspergillus niger, A. terreus, Cochliobolus specifer, Myrothecium verrucaria, Rhizoctonia solani, Spicaria fusispora, Penicillium sp., and Gliocladium sp.) were isolated from decomposing wheat straw and tested for their ability to utilize whole straw and its components, holocellulose (hemicellulose and cellulose) and cellulose, for the production of single-cell protein (SCP). It was found that C. specifer was the most efficient fungus for protein synthesis with the three substrates. Using potassium nitrate as N source in mixtures of 0.04 g N/g substrate (0.04% wt./vol.) at pH 4.5, it was found that incubation periods of 3, 4, and 5 days were optimal for protein production on cellulose and holocellulose fractions, and whole straw, respectively. Whole native straw was found to be the most recalcitrant to bioconversion into SCP; however, protein production was almost doubled when the lignin component was removed using a mixture of sodium chlorite and acetic acid.

Cellulose↗

Wheat embryo ribonucleates. V. Generation of N2--dimethylguanylate when 'fully sequenced' homogeneous species of transfer RNA are used as substrates for wheat embryo methyltransferases.

When S-adenosly[methyl-14-C]methionine and various species of transfer RNA are used as substrates for wheat embryo methyltransferases, the principal site of guanylate-N-2 methylation can be shown to be a G-residue between the stems of the dihydrouridine and anticodon loops. This common site of guanylate-N-2 methylation is referred to as the interstem target site. 2. When the interstem target site is the non-terminal G-residue in a G-C-G-C sequence, as in the cases of Escherichia coli tRNA1-Leu, tRNA-Ile, and tRNA3-Ser, there is preponderant dimethylation to yield N-2-dimethylguanylate. 3. When the interstem target site is part of a U-C-G-U sequence, as in the case of E. coli tRNAf-Met, there is diminished dimethylation and correspondingly increased monomethylation to yield N-2-monomethylguanylate. 4. When the interstem target site is the non-terminal G-residue in an A-U-G-G sequence, as in the case of yeast tRNA-Asp, there is negligible dimethylation and almost exclusive monomethylation to yield N-2-monomethylguanylate. 5. The concerted way in which the primary, secondary, and tertiary structures of tRNA molecules might influence the efficacy of these methylations is the subject of a brief discussion. Attention is also focused on the evolutionary and molecular basis for the generally non-random distributions of methylated oligonucleotide sequences in ribosomal and transfer ribonucleates.

Base Sequence↗

Exposure to ammoniated wheat straw as suckling calves improves performance of mature beef cows wintered on ammoniated wheat straw.

We studied how exposure to ammoniated wheat straw (AWS) early in life affected the performance of 32 mature crossbred beef cows (Bos taurus) (mean BW = 615 kg) wintered on AWS. Half (16) of these cows had been exposed as suckling calves to AWS for 66 d (Exposed), while the other 16 cows had no previous exposure to AWS (Naïve). Five years after the initial exposure, cows were stratified by BW and bred into 8 groups of 4 cows each. Exposed and Naïve cows occurred in each group, and groups of 4 cows were randomly assigned to one of eight pens. Cows were fed in these pens for a 150-d wintering period from December to May for 3 consecutive years. All cows were allowed ad libitum access to AWS and supplemented with alfalfa hay, vitamins, and minerals. Cows and their calves grazed irrigated meadow pastures for the remainder of the year. Cow BW and body condition score (BCS) were monitored monthly during the 3-yr study. Milk production was measured monthly from June to November of each year using the weigh-suckle-weigh technique. For all 3 yr, yearly average BW (P = 0.06, 0.03, 0.07) and BCS (P=0.07,0.001,0.01)were higher for Exposed than Naïve cows. Postpartum interval (PPI) to rebreeding, monitored using consecutive calving dates, was shorter for Exposed than Naïve cows during yr 1 and 2 (P = 0.004 and 0.02, respectively), but similar in yr 3 (P = 0.19). Exposed cows also produced more milk than Naïve cows during yr 1 and 2 (P = 0.04 and 0.07, respectively), but milk production was similar in yr 3 (P = 0.74). Collectively, calves exposed to AWS briefly early in life performed better as cows when reexposed to AWS from 5 to 8 yr later in life. Thus, researchers and managers should consider previous exposure to low-quality forages (LQF) when assigning cattle to studies involving the use of LQF or when considering using LQF to reduce food costs.

Ammonia↗

[Morphological variability of the wheat powdery mildew in the context of its parasitic adaptation to the lines of wheat Aegilops with different resistance].

We studied the interaction between the pathogen and wheat-Aegilops lines with different resistance as well as their parental forms in the course of powdery mildew infection using scanning electron microscopy. Line 51/99i proved to be similar to the parental form, Rodina variety, by the infection progress. The both genotypes featured pronounced adhesion of the primary infection structures to the surface of the plant epidermal cells in addition to formation of large developed colonies of the fungus, which indicates the partners compatibility. Development of powdery mildew on line 135/99i was similar to that on the parental form Aegilops speltoides K-389. The interaction between the partners featured decelerated incubation period. Rare parasite colonies formed from mycelial hyphae with multiple hyphal lobes and usually disturbed adhesion to the surface of the epidermal cells were observed. Such pattern of the pathogen development points to the resistance factors in these plant hosts, mostly, at the level of pathogen penetration. The types of resistance of lines 95/99i and 56/99i were not specific for the parental form Ae. speltoides K-389 but rather for Ae. speltoides specimens from other natural ranges (Ryabchenko et al., 2002). This suggests that Ae. speltoides has a polygenic immune potential with the elements inheritable by the hybrids irrespective of the used resistance donor.

Ascomycota↗

Homologies between members of the germin gene family in hexaploid wheat and similarities between these wheat germins and certain Physarum spherulins.

By screening approximately 10(6) plaques in a wheat DNA library with a "full-length" germin cDNA probe, two genomic clones were detected. When digested with EcoRI, one clone yielded a 2.8-kilobase pair fragment (gf-2.8) and the other yielded a 3.8-kilobase pair fragment (gf-3.8). By nucleotide sequencing, each of gf-2.8 and gf-3.8 was found to encode a complete sequence for germin and germin mRNA, and to contain appreciable amounts of 5'- and 3'-flanking sequences. The "cap" site in gf-2.8 was determined by primer extension and the corresponding site in gf-3.8 was deduced by analogy. The mRNA coding sequences in gf-2.8 and gf-3.8 are intronless and 87% homologous with one another. The 5'-flanking regions in gf-2.8 and gf-3.8 contain recognizable sites of what are probably cis-acting elements but there is otherwise little if any significant similarity between them. In addition to putative TATA and CAAT boxes in the 5'-flanking regions of gf-2.8 and gf-3.8, there are AT-rich inverted-repeats, GC boxes, long purine-rich sequences, two 19-base pair direct-repeat sequences in gf-2.8, and a remarkably long (200-base pair) inverted-repeat sequence (approximately 90% homology) in gf-3.8. An 8% difference between the mature-protein coding regions in gf-2.8 and gf-3.8 is reflected by a corresponding 7% difference between the corresponding 201-residue proteins. Most significantly, the same 8% difference between the mature-protein coding regions in gf-2.8 and gf-3.8 is allied with no change whatever in a central part (61-151) of the encoded polypeptide sequences. It seems likely that this central, strongly conserved core in the germins is of first importance in the biochemical involvements of the proteins. When an equivalence is assumed between like amino acids, the gf-2.8 and gf-3.8 germins show significant (approximately 44%) similarity to spherulins 1a and 1b of Physarum polycephalum, a similarity that increases to approximately 50% in the conserved core of germin. Near the middle (87-96) of the conserved core in the germins is a rare PH(I/T)HPRATEI decapeptide sequence which is shared by spherulins (1a and 1b) and germins (gf-2.8 and gf-3.8). These similarities are discussed in the context of evidence which can be interpreted to suggest that the biochemistry of germins and spherulins is involved with cellular, perhaps cell-wall responses to desiccation, hydration, and osmotic stress.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Mechanism of neutrophil chemiluminescence induced by wheat germ agglutinin: partial characterization of the antigens recognized by wheat germ agglutinin.

Wheat germ agglutinin (WGA) stimulated neutrophils to produce significant levels of luminol-dependent chemiluminescence (CL). Since WGA is known to bind N-acetylglucosamine (GlcNAc) oligomers and N-acetylneuraminic acid (NANA), we attempted to determine which binding property of WGA is essential for induction of CL. The succinylated form of WGA (SuWGA), which is no longer able to bind NANA, was still able to induce CL. N-Acetylglucosamine at a concentration of 20 mmol/L almost completely inhibited WGA-induced CL production by neutrophils, whereas bovine submaxillary gland mucin, a potent blocker of NANA binding of WGA, failed to inhibit CL production. Lectins with the GlcNAc-binding property were examined for their ability to induce CL. Those that have higher valences and have a tendency to bind GlcNAc oligomers in the internal portion of glycoconjugates were able to induce CL, whereas those that have low valences and bind terminal GlcNAc of glycoconjugates failed to induce CL even at high concentrations. Attempts were made to characterize the neutrophil membrane proteins recognized by WGA. Glycoproteins with a molecular weight of 25,000 daltons were identified by a 50 mmol/L GlcNAc elution of WGA gels loaded with 125I-labeled neutrophil membrane proteins. Elution with 500 mumol/L GlcNAc trimer produced several glycoproteins of different molecular weights in addition to the glycoproteins of 25,000 daltons. 125I-labeled WGA and SuWGA were used for autoradiographic analysis of cell extracts of the neutrophils separated on sodium dodecyl sulfate polyacrylamide gels. WGA recognized multiple glycoproteins of different molecular weights, whereas SuWGA bound only a few of them. Glycoproteins of 25,000 daltons, probably corresponding to those identified by 50 mmol/L GlcNAc elution, were also recognized.

Antigens↗