Allosteric activators of glucokinase: potential role in diabetes therapy

Glucokinase (GK) plays a key role in whole-body glucose homeostasis by catalyzing the phosphorylation of glucose in cells that express this enzyme, such as pancreatic beta cells and hepatocytes. We describe a class of antidiabetic agents that act as nonessential, mixed-type GK activators (GKAs) that increase the glucose affinity and maximum velocity (Vmax) of GK. GKAs augment both hepatic glucose metabolism and glucose-induced insulin secretion from isolated rodent pancreatic islets, consistent with the expression and function of GK in both cell types. In several rodent models of type 2 diabetes mellitus, GKAs lowered blood glucose levels, improved the results of glucose tolerance tests, and increased hepatic glucose uptake. These findings may lead to the development of new drug therapies for diabetes.     

Use of Alfalfa Hay, Compared to Feeding Practical Diets Containing Two Protein Levels, on Growth, Survival, Body Composition, and Processing Traits of Australian Red Claw Crayfish, Cherax quadricarinatus, Grown in Ponds

Juvenile red claw, Cherax quadricarinatus, were obtained from a commercial supplier and stocked into twelve 0.04‐ha ponds with an average water depth of approximately 1.1 m at a rate of 1100 red claw per pond (27,170 red claw/ha). Mean stocking weight (±SD) was 6.25 ± 3.0 g. Three replicate ponds were randomly assigned to each of the four treatments. In Treatment 1 (TRT1), red claw were stocked into ponds to which dried alfalfa hay (forage) was added at a rate of 500 kg/ha/mo; Treatment 2 (TRT2) consisted of red claw being fed a prepared diet containing 13% protein; in Treatment 3 (TRT3), red claw were fed a diet containing 13% protein at which had dried alfalfa hay been added at a rate of 500 kg/ha/mo; and in Treatment 4 (TRT4; control), red claw were fed a complete diet containing 28% protein. Water quality measurements were made three times weekly for dissolved oxygen and temperature (am , pm ), pH, total ammonia nitrogen, nitrite, and alkalinity averaging 7.17 mg/L, 8.96 mg/L, 25.5 C, 27.3 C, 8.68, 0.39 mg/L, 0.012 mg/L, and 106.5 mg/L, respectively. After 113 d, the final mean weight of red claw fed TRT3 was significantly (P < 0.05) higher (68.10 g) compared to that of red claw fed hay only (49.40 g) but not different from red claw fed TRT2 (56.03 g) and TRT4 (62.10 g). Likewise, red claw fed TRT3 had significantly higher percent weight gain (990%) compared to that of red claw fed hay only (690%) but not different from TRT2 (796%) and TRT4 (893%). Feed conversion ratio of TRT2, TRT3, and TRT4 and percent survival among all treatments did not differ significantly (P > 0.05), averaging 2.85 and 66.8%, respectively. Total yield of red claw fed TRT3 and TRT4 (968 and 952 kg/ha) was significantly (P < 0.05) higher compared to that of red claw fed hay only (617 kg/ha) but did not differ from red claw fed TRT2 (882 kg/ha). Tail meat yield of male red claw in TRT3 was significantly (P < 0.05) higher (17.3 g) compared to that of male red claw in TRT1 (12.7 g) and TRT2 (14.9 g) but not significantly different (P > 0.05) from male red claw in TRT4 (16.9 g). Tail meat yield of female red claw in TRT3 and TRT4 was significantly (P < 0.05) higher (14.2 and 13.9 g, respectively) compared to that of female red claw in TRT1 (10.5 g) and TRT2 (10.4 g). Tail muscle proximate compositions showed no significant differences (P > 0.05) in protein (wet weight basis) between males and females among treatments (diet) which averaged 16.2 and 14.6%; however, there were little to no significant differences in fat and ash between males and females among treatments. Results from this study indicate that pond‐cultured red claw stocked at 27,170 red claw/ha can be fed a practical diet containing 13% protein, with or without forage (alfalfa hay), compared to red claw fed a complete diet containing 28% protein; however, if alfalfa hay is added to the pond at 500 kg/ha/mo as the sole source of added nutrients, growth is reduced. Therefore, the use of alfalfa hay, in combination with a low‐protein pelleted diet may be a production method for pond‐grown red claw that may reduce costs for producers and thereby increase profits.     

Isolation and characterization of a bioactive mannose-binding protein from the Chinese chive Allium tuberosum

A mannose-binding protein was isolated from two different cultivars of the Chinese chive Allium tuberosum by extraction with 0.2 M NaCl, ammonium sulfate precipitation, and affinity chromatography on mannose agarose and fetuin agarose. It exhibited hemagglutinating activity toward rabbit erythrocytes. The lectin (agglutinin) was adsorbed on the mannose-agarose column, but not on the fetuin-agarose column. This A. tuberosum lectin (ATL) is unglycosylated, and not sialic acid binding. Lectins isolated from the two cultivars exhibited the same molecular mass of 25 kDa on gel filtration (Superose 12) and 12.5 kDa on SDS-polyacrylamide gel electrophoresis, indicating that they might be a dimeric protein composed of two identical subunits. The N-terminal amino acid sequence analysis of the lectin of various cultivars of A. tuberosum revealed that they were identical and showed 50%, or more, homology to the lectins from Galanthus nivalis (family Amaryllidaceae), Narcissus tazetta (family Amaryllidaceae), and Aloe arborescenes (family Liliaceae).     

Extraction,semi-quantification,and fast on-line identification of oligopeptides in Grana Padano cheese by HPLC-MS

Sixteen samples of Grana Padano cheese aged from 2 to 33 months were analyzed by HPLC-MS. The extraction process involved the use of diluted HCl, thus avoiding a strong deproteinizing agent (TCA), and allowing to maintain in solution also very lipophilic peptides. The molecular mass of the most abundant peptides were determined by electrospray ionization (ESI) mass spectrometry. A new method was developed based on the small fragmentation peaks arising from in-source fragmentation and from software analysis of the known casein sequences, which in many cases allowed the direct on-line identification of the oligopeptide sequences. Several new peptides never previously reported were identified, some of which containing bioactive sequences, consistently with what was described in the literature. Semiquantification of peptides at the different stages of aging was also performed by using a suitable internal standard, providing new insights into the evolution of the oligopeptide fraction during aging.     

Preliminary studies for the differentiation of apple juice samples by chemometric analysis of solid-phase microextraction-gas chromatographic data

A combination of gas chromatography (GC) and chemometrics was evaluated for its ability to differentiate between apple juice samples on the basis of apple variety and applied heat treatment. The heat treatment involved exposure of 15 mL juice samples for 30 s in a 900 W domestic microwave oven. The chromatographic results were subjected to two chemometric procedures: (1) partial least squares (PLS) regression and (2) linear discriminant analysis (LDA) applied to principal component (PC) scores. The percent correct classification of samples were obtained from PLS and LDA in terms of separation on the basis of apple variety and applied heat treatment. PLS gave the highest level of correct classification of the apple juice samples according to both variety and heat treatment, 92.5% correct classification in each case. When LDA was performed on the PC scores obtained from GC analysis, 87.5% and 80% of samples were correctly classified according to apple variety used and applied heat treatment, respectively.     

In vitro fermentability of differently substituted xylo-oligosaccharides

Xylo-oligosaccharides (XOS) with various substituents were fermented in vitro by fecal inocula (FI) from four human volunteers to study the influence of substitution on the ability and rate of fermentation and on the production of short-chain fatty acids (SCFA) and lactate. By all FI used nonsubstituted XOS (nXOS) and arabino-XOS (AXOS) were fermented more quickly than the more complex structures of acetylated XOS (AcXOS) and XOS containing a 4-O-methylglucuronic acid group (GlcA(me)XOS). In the first stage (0-40 h) of the fermentations of nXOS and AXOS mainly acetate and lactate were formed. The fermentations of AcXOS and GlcA(me)XOS resulted in a lower lactate production, whereas the concentration of propionate and butyrate increased. These results put emphasis on the detailed elucidation of the structural features of nondigestible oligosaccharides in general to understand their fermentation mechanisms more precisely.     

Antimicrobial and antioxidant effects of milk protein-based film containing essential oils for the preservation of whole beef muscle

Milk protein-based edible films containing 1.0% (w/v) oregano, 1.0% (w/v) pimento, or 1.0% oregano-pimento (1:1) essential oils mix were applied on beef muscle slices to control the growth of pathogenic bacteria and increase the shelf life during storage at 4 degrees C. Meat and film were periodically tested during 7 days for microbial and biochemical analysis. The lipid oxidation potential of meat was evaluated by the determination of thiobarbituric reactive substances (TBARS). The availability of phenolic compounds from essential oils was evaluated by the determination of total phenolic compounds present in the films during storage. Antioxidant properties of films during storage were also evaluated following a modified procedure of the N,N-diethyl-p-phenylenediamine colorimetric method. Oregano-based films stabilized lipid oxidation in beef muscle samples, whereas pimento-based films presented the highest antioxidant activity. The application of bioactive films on meat surfaces containing 10(3) colony-forming units/cm2 of Escherichia coli O157:H7 or Pseudomonas spp. showed that film containing oregano was the most effective against both bacteria, whereas film containing pimento oils seems to be the least effective against these two bacteria. A 0.95 log reduction of Pseudomonas spp. level, as compared to samples without film, was observed at the end of storage in the presence of films containing oregano extracts. A 1.12 log reduction of E. coli O157:H7 level was noted in samples coated with oregano-based films.     

Assessing limitations on population growth in two critically endangered Acacia taxa

Flowering phenology, soil seedbank and the impact of fire, weeds and grazing were investigated in two rare Acacia taxa restricted to the agricultural district north of Perth Western Australia. Acacia aprica is known from six extant populations and one extinct population. Five of the extant populations are restricted to linear road reserves with the sixth population located in a small native vegetation remnant. Acacia cochlocarpa ssp. cochlocarpa is known from a single population also situated on a road verge. Size class structure, levels of canopy death and an absence of juveniles indicated that all populations are in decline. Flowering intensity and success varied between populations and years in A. aprica and between years in A. cochlocarpa ssp. cochlocarpa. Seed bank analysis indicated that seeds were patchily distributed while experimental fires demonstrated that such events could break seed dormancy and promote germination. Both taxa have similar germination physiologies and showed increased germination after seeds were exposed to heat. Following emergence, however, competition with annual weeds had a negative impact on seedling growth and survival in both taxa. While vertebrate grazing had some influence, weeds were the major inhibitory influence on recruitment. Reduced fire frequencies since fragmentation may be responsible for population decline in both taxa but other site specific factors such as weeds and grazing may affect the establishment of seedlings following fire.     

Oxalate and phytate concentrations in seeds of soybean cultivars [Glycine max (L.) Merr.

This study analyzed soybean seeds from 116 cultivars for total, insoluble, and soluble oxalate (Ox), phytate (InsP6), calcium (Ca), and magnesium (Mg) because of their potential beneficial or harmful effects on human nutrition. These cultivars were divided into four groups (A-D) on the basis of the year and geographic location where they were grown. Oxalate concentration ranged from about 82 to 285 mg/100 g of dry seed. The InsP6 concentration ranged from 0.22 to 2.22 g/100 g of dry seed. There was no correlation between Ox and InsP6 within or among the four groups of cultivars. There was a significant correlation between total Ox and Ca, but not Mg, in group D cultivars (r = 0.3705; p < 0.0005). No significant relationship was found in the group A-C cultivars. Eleven group D cultivars had InsP6 less than 500 mg/100 g, but all had total Ox of 130 mg/100 g or greater. Five cultivars from groups A-C had relatively low InsP6 (group B; < or =1.01 g/100 g) and low Ox (<140 mg/100 g). These cultivars could be useful for producing soy foods beneficial to populations at risk for kidney stones and for improved mineral bioavailability. The Ox and InsP6 concentrations of the cultivars indicate that choosing specific parents could generate seeds in succeeding generations with desirable Ox and InsP6 concentrations.