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Dietary fat intake and regulation of energy balance: implications for obesity.

Obesity represents a major threat to health and quality of life. Although obesity has strong genetic determinants, the increasing prevalence of obesity in populations around the world suggests that environmental factors are promoting or exacerbating the problem. Experts are calling for public health efforts to deal with the global epidemic of obesity. Such a campaign would require that we identify and modify environmental factors that promote obesity. Our current food supply is high in fat, and high fat diets have been suggested to promote obesity by increasing energy intake, thus increasing the probability of positive energy balance and weight gain. However, others argue that high fat diets are not promoting obesity. In this paper, we review evidence from animal studies, carefully controlled laboratory studies, cross-sectional studies, clinical trials and studies in individuals at high risk to develop obesity. Although there are many environmental factors promoting excess energy intake and discouraging energy expenditure, it is clear that consumption of a high fat diet increases the likelihood of obesity and that the risk of obesity is low in individuals consuming low fat diets. On the basis of the available data, the current public health recommendations to lower dietary fat intake appear to be appropriate.

Animals↗

Olfactory conditioning of proboscis activity in Drosophila melanogaster.

Olfactory learning and memory processes in Drosophila have been well investigated with aversive conditioning, but appetitive conditioning has rarely been documented. Here, we report for the first time individual olfactory conditioning of proboscis activity in restrained Drosophila melanogaster. The protocol was adapted from those developed for proboscis extension conditioning in the honeybee Apis mellifera. After establishing a scale of small proboscis movements necessary to characterize responses to olfactory stimulation, we applied Pavlovian conditioning, with five trials consisting of paired presentation of a banana odour and a sucrose reward. Drosophila showed conditioned proboscis activity to the odour, with a twofold increase of percentage of responses after the first trial. No change occurred in flies experiencing unpaired presentations of the stimuli, confirming an associative basis for this form of olfactory learning. The adenylyl cyclase mutant rutabaga did not exhibit learning in this paradigm. This protocol generated at least a short-term memory of 15 min, but no significant associative memory was detected at 1 h. We also showed that learning performance was dependent on food motivation, by comparing flies subjected to different starvation regimes.

Adenylyl Cyclases↗

Heat-stable phytases in transgenic wheat (Triticum aestivum L.): deposition pattern, thermostability, and phytate hydrolysis.

The present paper addresses the question of thermotolerance of in planta synthesized heterologous enzymes using phytase as a model. Two individual transgenic wheat materials expressing an Aspergillus fumigatus phytase with a low denaturation temperature (62.5 degrees C) but a high refolding capacity, and a rationally designed consensus phytase engineered to a high denaturation temperature (89.3 degrees C), were evaluated. High levels of endosperm specific expression were ensured by the wheat high molecular weight glutenin 1DX5 promoter. Immunodetection at the light and electron microscopical level shows unequivocally that the heterologous phytase is deposited in the vacuole, albeit that the transformation constructs were designed for secretion to the apoplast. Evaluation of heat stability properties and kinetic properties unraveled that, under these deposition conditions, heat stability based on high unfolding temperature is superior to high refolding capacity and represents a realistic strategy for improving phosphate and mineral bioavailability in cereal-based feed and food.

6-Phytase↗

Effect of genetically modified, low-phytic acid maize on absorption of iron from tortillas.

BACKGROUND: Genetically modified, low-phytic acid strains of maize were developed to enhance mineral absorption, but have not been tested previously in humans. OBJECTIVES: We evaluated the mineral and phytic acid contents of a low-phytic acid "flint" maize (LPM, the lpa-1-1 mutant) and its parent, wild-type strain (WTM) and measured iron absorption from tortillas prepared with each type of maize and from a reference dose of ferrous ascorbate. DESIGN: Proximate composition and mineral and phytic acid contents were measured by standard techniques. Iron absorption from tortillas was evaluated by using the extrinsic tag method and was measured as the incorporation of radiolabeled iron into the red blood cells of 14 nonanemic men 2 wk after intake. RESULTS: The phytic acid content of LPM was 3.48 mg/g, approximately 35% of the phytic acid content of WTM; concentrations of macronutrients and most minerals were not significantly different between strains. Iron absorption results were adjusted to 40% absorption of ferrous ascorbate. Iron absorption was 49% greater from LPM (8.2% of intake) than from WTM (5.5% of intake) tortillas (P < 0.001, repeated-measures analysis of variance). CONCLUSION: Consumption of genetically modified, low-phytic acid strains of maize may improve iron absorption in human populations that consume maize-based diets.

Adult↗

Pseudomonas lipases: molecular genetics and potential industrial applications.

Lipases are esterases able to hydrolyze water-insoluble esters such as long-chain triglycerides. These enzymes also catalyze the formation of esters (esterification) and the exchange of ester bonds (transesterification) when present in nonaqueous media. Lipases display a high degree of specificity and enantioselectivity for esterification and transesterification reactions, and thus their potential uses in industry are very wide. These potential industrial applications have been an important driving force for lipase research during the last several years, and in particular for the study of lipases produced by microorganisms. Pseudomonas lipases are very interesting because they display special biochemical characteristics not common among the lipases produced by other microorganisms, such as their thermoresistance and activity at alkaline pHs. Recently, several Pseudomonas genes have been cloned and sequenced, and the regulation of their expression is beginning to be understood. The molecular genetic approach to the study of Pseudomonas lipases will permit the construction of recombinant strains with increased lipase productivity and will provide the opportunity to modify these enzymes to suit particular industrial applications.

Biotechnology↗

Drosophila short neuropeptide F regulates food intake and body size.

Neuropeptides regulate a wide range of animal behavior including food consumption, circadian rhythms, and anxiety. Recently, Drosophila neuropeptide F, which is the homolog of the vertebrate neuropeptide Y, was cloned, and the function of Drosophila neuropeptide F in feeding behaviors was well characterized. However, the function of the structurally related short neuropeptide F (sNPF) was unknown. Here, we report the cloning, RNA, and peptide localizations, and functional characterizations of the Drosophila sNPF gene. The sNPF gene encodes the preprotein containing putative RLRF amide peptides and was expressed in the nervous system of late stage embryos and larvae. The embryonic and larval localization of the sNPF peptide in the nervous systems revealed the larval central nervous system neural circuit from the neurons in the brain to thoracic axons and to connective axons in the ventral ganglion. In the adult brain, the sNPF peptide was localized in the medulla and the mushroom body. However, the sNPF peptide was not detected in the gut. The sNPF mRNA and the peptide were expressed during all developmental stages from embryo to adult. From the feeding assay, the gain-of-function sNPF mutants expressed in nervous systems promoted food intake, whereas the loss-of-function mutants suppressed food intake. Also, sNPF overexpression in nervous systems produced bigger and heavier flies. These findings indicate that the sNPF is expressed in the nervous systems to control food intake and regulate body size in Drosophila melanogaster.

Amides↗

Transgenic-Bt potato plant resistance to the colorado potato beetle affect the aphid parasitoid Aphidius nigripes.

Using the biotechnological plant resistance for herbivore control with less reliance on chemicals in integrated pest management (IPM) programs critically depends on predictable interactions with no-target organisms of various trophic levels. Plant resistance to insect pests based on recombinant Bacillus thuringiensis could interfere with natural enemies of non target pests. Performance of the potato aphid parasitoid Aphidius nigripes was studied on the 'Superior-BT line transgenic for the CryllIA toxin of B. thuringiensis, resistance to the Colorado potato beetle; and none transformed 'Superior' line which served as control. Parasitoid survival was significantly lower on the 'Superior-BT' line compared to control. Adult females were largest on 'Superior' and smallest on BT potatoes. This difference was reflected on parasitoid fecundity, which was lowest on 'Superior-BT', and highest on Superior. The results indicate that factor of potato resistance to the Colorado potato beetle affected the fitness of a parasitold of the aphid Macrosiphum euphorbiae, a secondary pest of potato. The effects on the parasitoid were complex but were generally interpretable in terms of host aphid quality variation among potato lines used as food by the aphids during parasitoid development.

Animals↗

High level expression of a recombinant acid phytase gene in Pichia pastoris.

AIMS: To achieve high phytase yield with improved enzymatic activity in Pichia pastoris. METHODS AND RESULTS: The 1347-bp phytase gene of Aspergillus niger SK-57 was synthesized using a successive polymerase chain reaction and was altered by deleting intronic sequences, optimizing codon usage and replacing its original signal sequence with a synthetic signal peptide (designated MF4I) that is a codon-modified Saccharomyces cerevisiae mating factor alpha-prepro-leader sequence. The gene constructs containing wild type or modified phytase gene coding sequences under the control of the highly-inducible alcohol oxidase gene promoter with the MF4I- or wild type alpha-signal sequence were used to transform Pichia pastoris. The P. pastoris strain that expressed the modified phytase gene (phyA-sh) with MF4I sequence produced 6.1 g purified phytase per litre of culture fluid, with the phytase activity of 865 U ml(-1). The expressed phytase varied in size (64, 67, 87, 110 and 120 kDa), but could be deglycosylated to produce a homogeneous 64 kDa protein. The recombinant phytase had two pH optima (pH 2.5 and pH 5.5) and an optimum temperature of 60 degrees C. CONCLUSIONS: The P. pastoris strain with the genetically engineered phytase gene produced 6.1 g l(-1) of phytase or 865 U ml(-1) phytase activity, a 14.5-fold increase compared with the P. pastoris strain with the wild type phytase gene. SIGNIFICANCE AND IMPACT OF THE STUDY: The P. pastoris strain expressing the modified phytase gene with the MF4I signal peptide showed great potential as a commercial phytase production system.

6-Phytase↗

New variant Creutzfeldt-Jakob disease and bovine spongiform encephalopathy.

New variant Creutzfeldt-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE) are invariably fatal, subacute degenerative diseases of the brain that are classified as transmissible spongiform encephalopathies. BSE was first diagnosed in 1986 as part of an ongoing epizootic in the United Kingdom that was amplified by the feeding of rendered bovine meat-and-bone meal to young calves. As of June 1997, a total of 17 cases of new variant CJD have been reported among residents of the United Kingdom, an increase of seven cases since March 1996, when concern was first expressed about the possible emergence of new variant CJD as a novel epidemic caused by the spread of BSE to humans. Accumulating experimental and epidemiologic data support this concern and have led the CDC to support the Food and Drug Administration's efforts to modify the protein content of ruminant feed in the United States as a prudent measure to protect the health of animals and the public.

Animals↗

Transgenic plants as vaccine production systems.

Transgenic plants that express foreign proteins with industrial or pharmaceutical value represent an economical alternative to fermentation-based production systems. Specific vaccines have been produced in plants as a result of the transient or stable expression of foreign genes. It has recently been shown that genes encoding antigens of bacterial and viral pathogens can be expressed in plants in a form in which they retain native immunogenic properties. Transgenic potato tubers expressing a bacterial antigen stimulated humoral and mucosal immune responses when they were provided as food. These results provide 'proof of concept' for the use of plants as a vehicle to produce vaccines.

Animals↗

Biochemical safety evaluation of transgenic rice seeds expressing T cell epitopes of Japanese cedar pollen allergens.

UNLABELLED: Transgenic rice seeds, which express a hybrid peptide comprising seven predominant human T cell epitopes (7Crp) derived from Japanese cedar pollen allergens, have been shown to function as an effective edible vaccine for the control of pollen allergen-induced responses. In this study, we characterized biochemical properties of transgenic seeds expressing the 7Crp peptide. The levels of chemical compositions, such as carbohydrate, protein, lipid, amino acid, fatty acid, mineral, and vitamin, were substantially equivalent between transgenic 7Crp and its nontransgenic counterpart seeds. The contents of three major allergenic proteins in transgenic seeds were not enhanced by expression of the 7Crp peptide when compared with those of nontransgenic seeds. The 7Crp peptide expressed in seeds was susceptible to simulated gastric/intestinal fluids. N-Glycosylation was not observed in the 7Crp peptide sequence. These results indicate that transgenic 7Crp seeds are substantially equivalent to nontransgenic parental seeds except for the presence of the 7Crp peptide. KEYWORDS: Food safety assessment; transgenic rice seed; edible vaccine; peptide-based immunotherapy; Japanese cedar pollinosis.

Allergens↗

Effect of selection for improved body composition on muscle and meat characteristics of broilers from experimental and commercial lines.

Meat quality (pH, color, and drip loss) and muscle characteristics (composition and metabolic pattern) were compared among four broiler lines: an experimental line, a commercial line selected for increased body weight and breast yield, and the respective unselected control lines. By comparison to their controls, the commercially selected birds exhibited higher body weight and breast yield (127 and 61%, respectively), whereas only breast yield was increased (21%) in the experimental selected line. Commercial selection resulted in higher protein content and lower moisture in the breast muscle. Experimental and commercial selection decreased the heme pigment content, which could explain why breast meat of the selected birds was more pale and less red. This change in the color did not correspond to a pale, soft, and exudative (PSE) defect, as the selected birds did not exhibit excessive drip loss. By comparison with their controls, both selected lines showed a lower rate and extent of pH decline postmortem, which was consistent with the lower glycolytic potential they also exhibited. However, no significant changes in the metabolic pathways of breast muscle, as measured by enzyme activities, could be found to explain differences in rates of pH decline among lines. This study did not support the idea that selection had a negative impact on meat quality, despite evidence of modified breast metabolism.

Animals↗

Attentional set-shifting in mice: modification of a rat paradigm, and evidence for strain-dependent variation.

Increasingly precise molecular genetic tools are available to study in mice the cellular mechanisms underlying complex brain functions, but the behavioural paradigms to assess these functions often lack the required specificity. In this study, an attentional set-shifting paradigm to assess medial frontal cortex functions in rats was modified for use in mice and variation between two relevant mouse strains assessed. Male 129/SvEv and C57BL/6J mice and their F1 intercross (n=8 per genotype) were trained to dig in bowls for a food reward. On four consecutive days, mice performed a series of discriminations to criterion (six consecutive correct choices) between pairs of food bowls that differed along two dimensions (odour, digging medium), including a reversal, an intra-dimensional shift, and an extra-dimensional shift. Mice from the 129 strain performed significantly better than C57 mice in the initial acquisition of a simple discrimination and in the final extra-dimensional shift test, with no difference in the reversal and intra-dimensional shift. Performance of the F1 mice was intermediate or similar to that of the 129 mice. These results indicate a selective difference between these two strains in attentional selection processes that have been shown in humans, monkeys and rats to be mediated by prefrontal cortex.

Animals↗

[The risk of transmission of salmonellae in poultry farming: detection and prevention in Europe].

While salmonellas can cause disease problems among poultry, they remain essentially a concern for public health, as a cause of outbreaks of food poisoning. The principal site of multiplication of these bacteria is the digestive tract, particularly the caecum, which may result in widespread contamination of the environment. The pathogenicity of salmonellas depends on the invasive properties and the ability of the bacteria to survive and multiply within cells, particularly macrophages. These properties are the source of vertical transmission which, in the case of survival of the embryo, can result in contamination of a flock or, in the case of embryonic mortality, can result in an explosion of contaminated eggs. Salmonella infection can be diagnosed by isolating the bacteria and/or serological testing of the flock. European Union Directive 92/117/EC, modified by Directive 97/22/EC, stipulates either the destruction of infected flocks of breeding birds, or decontamination of the flock in an effective way, before normal trade in products can be resumed. Noteworthy examples of effective measures suitable for prophylaxis of Salmonella infection in poultry flocks include the slaughter of infected breeding stock, the creation of sanitary barriers at building entrances, heat treatment of feed, the use of competitive exclusion, selection of breeds genetically resistant to Salmonella, and occasional vaccination and antibiotic treatment. However, the most effective means of reducing food poisoning remains adequate cooking of food and maintenance of the cold chain.

Animals↗

The deadly sins and diabetes.

Type 2 diabetes is a spectacularly large and growing health problem. Patients with diabetes have healthcare expenditures that are twice those of the average citizen. We do not clearly understand the genetic predisposition that allows type 2 diabetes to develop, but we do know that lifestyle factors are the most important modifiable factors leading to that disorder. A proper diet and regular exercise (manifested by the presence of a healthy body mass index) can prevent development of latent diabetes, or reverse the manifestations of overt diabetes. It is unlikely that the food industry-already bloated by the high profits from enticing people to buy and consume more and more calories-will voluntarily decide to limit its income. So we will need a number of initiatives from healthcare professionals and from governments at all levels of society to turn this situation around. Some of the measures will require enormous political courage and foresight on the part of our elected officials. Without that, we will hardly need to worry about defense from external enemies. As a nation, we will soon be so fat that we will not be able to waddle to work or to war; those who can will be trying to earn enough money to pay for all the glucose meters and insulin needles and hypoglycemic, antihypertensive, and cholesterol-lowering drugs "required" for the evidence-based treatment of hundreds, even thousands, of blind, neuropathic amputees crowding our cardiac care and dialysis units.

Behavior Therapy↗

Quality and safety evaluation of genetically modified potatoes spunta with Cry V gene: compositional analysis, determination of some toxins, antinutrients compounds and feeding study in rats.

The aim of this study was to evaluate the composition, nutritional and toxicology safety of GM potato Spunta lines compared to that of conventional potato Spunta. Compositional analyses were conducted to measure the proximate chemical composition with references to 14 components, total solid, protein, lipid, crude fibre, ash, carbohydrate, starch, reducing sugar, nonreducing sugar, sodium, calcium, potassium, phosphorus, and ascorbic acid. Some toxins and anti-nutrients compounds were determined. Feeding study of GM potatoes line (G2 and G3) in rats were done for 30 days. Four groups of albino rats were used for studying the effect and the safety assessment of GM potatoes Spunta G2 and G3. Group (I) was fed on control basal diet, group (II) was fed on control diet plus 30% freeze-dried nongenetically modified potato Spunta, group (III) was fed on control diet plus 30% freeze-dried genetically modified potato Spunta, and group (IV) was fed on control diet plus 30% freeze-dried genetically modified potato Spunta GMO G3. There were no significant differences between GM potatoes G2, G3, and Spunta control potato line in the proximate chemical composition. The levels of glycoalkaloids in transgenic potato tubers and nontransgenic were determined and there were also no significant differences between the GM potatoes and conventional potato line, the levels were in agreement with a safety level recommended by FAO/WHO (200 mg/ kg) for acute toxicity. Protease inhibitor activity and total phenol were estimated and no significant differences between the GM potatoes line and conventional potato Spunta line were found. During the period tested, rats in each group (I, II, III, IV) grew well without marked differences in appearance. No statistical difference were found in food intake, daily body weight gain and feed efficiency. But there is a slightly significant difference in finally body weight between the control group and experimental groups. No significant difference were found in serum biochemical value between each groups, and also between relative organs weight (liver, spleen, heart, kidney, testes). From these results, it can be concluded that the GM potatoes Spunta line (G2 and G3) with Cry V gene are confirmed to have nearly the composition and biochemical characteristics as non-GM potato Spunta.

Amino Acids↗

Role of antimutagens/anticarcinogens in cancer prevention.

Recently it has become increasingly clear that chemicals found in our foods and beverages can prevent the genetic damage that leads to cancer initiation. Such substances may also affect subsequent events in the pathways that lead to cancer, and may have the potential to inhibit the mutations that allow tumor cells to become resistant to antitumor agents. We describe here the antimutagenic potential of Glabrene analogs against EMS-induced mutations utilizing modified Ames tests in S. typhimurium TA 100 and E. coli JC 5088. Results of studies of the ability of well-known antioxidants such as EGCG and related compounds to prevent drug resistance mutations in microorganisms are described, and their possible significance in the prevention of chemotherapeutic drug-resistance in tumor cells is discussed.

Anticarcinogenic Agents↗

Novel lantibiotics and their pre-peptides.

In recent years there has been a considerable increase in studies of bactericidal peptides produced by Gram-positive bacteria, with particular emphasis upon their potential application as food preservatives. A number of these peptides contain lanthionine and other post-translationally modified amino acid residues. The lanthionine-containing molecules (lantibiotic) appear to have evolved in two quite different lineages, type A and type B. This mini-review introduces the reader to several of the more recently described type A lantibiotics for which relatively detailed biochemical and/or genetic data has already been gathered. A wider diversity of compounds of type A lantibiotics has been described in the recent years. Novel features of some of the more recently described type A lantibiotics to be reported in this review include: a) New modifications such as D-Ala and 2-hydroxypropionyl residues, both derived from serine. b) Different types of pre-lantibiotic leader sequences. c) The apparent requirement for different numbers and types of genes for synthesis of some active type A lantibiotics. d) Cytolysin functions as both a hemolysin and a bacteriocin. e) One of the newly-described lantibiotics (lactocin S) does not have any net charge at neutral pH another (carnocin UI49) is the largest of the lantibiotics discovered and the killing action of another (cytolysin) has been shown to be depend on the interaction of two peptides.

Alanine↗