Biotin serves as a covalently bound coenzyme in five human carboxylases; biotin is also attached to histones H2A, H3, and H4, although the abundance of biotinylated histones is low. Biotinylation of both carboxylases and histones is catalyzed by holocarboxylase synthetase. Human biotin requirements are unknown. Recommendations for adequate intake of biotin are based on the typical intake of biotin in an apparently healthy population, which is only a crude estimate of the true intake due to analytical problems. Importantly, intake recommendations do not take into account possible effects of biotin deficiency on impairing genome stability. Recent studies suggest that biotin deficiency causes de-repression of long terminal repeats, thereby causing genome instability. While it was originally proposed that these effects are caused by loss of biotinylated histones, more recent evidence suggests a more immediate role of holocarboxylase synthetase in forming multiprotein complexes in chromatin that are important for gene repression. Holocarboxylase synthetase appears to interact physically with the methyl-CpG-binding domain protein 2 and, perhaps, histone methyl transferases, thereby creating epigenetic synergies between biotinylation and methylation events. These observations might offer a mechanistic explanation for some of the birth defects seen in biotin-deficient animal models.
Two groups of isoforms of rhEPO, at a concentration of 300 µg/ml, were tested as putative inhibitors of the lectinic hemagglutination reaction in order to obtain affinity ligand(s) for hormone purification: groups I (pI: 3.80; 3.89; 3.95; 4.07, 4.15 and 4.26) and groups II (pI: 4.15, 4.26; 4.38; 4.51; 4.72 and 4.93) Crude extracts from the vegetable materials Abrus precatorious (Abrin), Artocarpus incisa (Frutalin), Artocarpus integrifolia (Jacalin), Canavalia ensiformes (ConA), Canavalia brasiliensis (Conbr), Cratylia floribunda, Dioclea altissima (DAL), Dioclea grandiflora (DGL), Erythrina vellutina (EVL), Erythrina cristagalli, Lutaelburgia auriculata (lectin not fully characterized yet), Lycopersicum esculentum (LEA), Phaseolus vulgaris (PHA), Ricinus communis (Ricin) and Triticum vulgaris (WGA) were used. Only some of the galactose-specific lectins and the GlcNAc-specific lectins showed rapid full inhibition of the hemagglutination reaction for the less acidic isoforms and the total isoforms of rhEPO, respectively. On this basis, the selected lectins were purified by affinity chromatoghraphy and covalently coupled to cyanogen bromide activated Sepharose® (Amersham-Pharmacia). CHO.K1 cell culture supernatant containing rhEPO was loaded onto the lectin resins and the recoveries were calculated by using specific elutions
Among the Bauhinia species, B. cheilantha stands out for its seed protein content. However, there is no record of its nutritional value, being used in a nonsustainable way in the folk medicine and for large-scale extraction of timber. The aim of this study was to investigate the food potential of B. cheilantha seeds with emphasis on its protein quality to provide support for flora conservation and use as raw material or as prototype for the development of bioproducts with high added socioeconomic value. B. cheilantha seeds show high protein content (35.9%), reasonable essential amino acids profile, low levels of antinutritional compounds, and nutritional parameters comparable to those of legumes widely used such as soybean and cowpea. The heat treatment of the seeds as well as the protein extraction process (to obtain the protein concentrate) increased the acceptance of diets by about 100% when compared to that of raw Bc diet. These wild legume seeds can be promising alternative source of food to overcome the malnutrition problem faced by low income people adding socioeconomic value to the species.
About 40% of the hotspots for meiotic recombination contain the degenerate consensus sequence 5’-CCNCCNTNNCCNC-3’. Here we present a novel protocol for enriching hotspot sequences from digested genomic DNA by using biotinylated oligonucleotides and streptavidin-coated magnetic beads. The captured hotspots can be released by simple digestion with restriction enzymes for subsequent characterization by second generation sequencing or PCR. The capture protocol specifically enriches hotspot sequences, judged by using fluorophore-conjugated synthetic oligonucleotides and synthetic double-stranded oligonucleotides in combination with PCR. The capture protocol enriches single stranded DNA, denatured double-stranded DNA, and large fragments (>3,000 bp) of digested plasmid DNA with good efficacy. No false positive and false negatives were detected when enriching digested DNA from human cell cultures and primary human cells. The protocol can probably be adapted to enriching sequences other than the hotspot sequence by altering the sequence in the capture oligonucleotide. We intend to apply this protocol in studies assessing effects of micronutrient status on meiotic recombination events in human sperm.
The acute effects of hyperglycemia in cognitive function and mood are not well elucidated. The aim of this study was to evaluate whether acute hyperglycemia alters the exploratory behavior of adult zebrafish. Fish were maintained aquariums containing 55 mM D-glucose solution for 14 days. Blood glucose levels and behavioral analysis were assessed before and after the D-glucose challenge. Immersion of adult zebrafish in a 55 mM D-glucose solution (14 days) increased blood glucose levels until the 14th day. There was no difference in weight gain between the two groups. Hyperglycemia did not induce anxiogenic or anxioly-tic behavior, nor did it alter the locomotor activity of the animals. We conclude that acute hyperglycemia does not alter the exploratory behavior of the adult zebrafish.
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