Detection of anisakids in fish and seafood products by real-time PCR

Anisakids are a group of widely distributed nematodes which have acquired high social relevance due to their involvement in foodborne infections caused by consumption of raw or undercooked seafood. A TaqMan^®-LNA probe real-time assay targeting the cytochrome oxidase subunit I (COI) was developed allowing the simultaneous detection of the most important anisakids species present in fish and seafood products.The determination of the detection limit in terms of ppm was 1 ppmFor the validation of method developed, twenty fish and cephalopod samples were experimentally contaminated with anisakid. It was checked that in cases in any anisakids species was present, it was detected because the Ct was always less than 35 and did not produce any case false negatives. The main novelty of this work lies in the fact that it can be applied to all kinds of processed products, including those undergoing intensive processes of transformation, as for instance canned foods. The proposed methodology is rapid, robust, highly sensitive and readily adaptable in routine molecular diagnostic laboratories, and can be employed as molecular screening method in order to assess the food security.     

Invertase activity in fresh and processed honeys

The enzymatic activities of 147 samples of commercially produced Spanish polyfloral and monofloral honeys are reported. Important variations in invertase activity and peroxide accumulation were determined to obtain objective information related to honey quality. Variations from 4.04 (Robinia pseudoacacia) to 25.61 g sucrose hydrolysed per 100 g h⁻¹ (SN) (Castanea sativa) and from 11.31 (Erica vagans) to 45.25 µg H₂O₂ g⁻¹ h⁻¹ (Hedysarum coronarium) were detected, with a range from 10.57 to 46.2 SN in polyfloral honeys. Honeys from Erica spp had different invertase activities—high for E cinerea (18.3 SN) and low for E vagans (8.36 SN). The different heating processes of honey have been evaluated to determine the best treatment for preserving invertase activity. The hydroxymethylfurfural content is not a sufficiently good indicator of the degree of deterioration of honey. Another objective was to assess the use of invertase activity in honey quality legislation. © 2000 Society of Chemical Industry     

A case-based approach for coordinated action selection in robot soccer

Designing coordinated robot behaviors in uncertain, dynamic, real-time, adversarial environments, such as in robot soccer, is very challenging. In this work we present a case-based reasoning approach for cooperative action selection, which relies on the storage, retrieval, and adaptation of example cases. We focus on cases of coordinated attacking passes between robots in the presence of the defending opponent robots. We present the case representation explicitly distinguishing between controllable and uncontrollable indexing features, corresponding to the positions of the team members and opponent robots, respectively. We use the symmetric properties of the domain to automatically augment the case library. We introduce a retrieval technique that weights the similarity of a situation in terms of the continuous ball positional features, the uncontrollable features, and the cost of moving the robots from the current situation to match the case controllable features. The case adaptation includes a best match between the positions of the robots in the past case and in the new situation. The robots are assigned an adapted position to which they move to maximize the match to the retrieved case. Case retrieval and reuse are achieved within the distributed team of robots through communication and sharing of own internal states and actions. We evaluate our approach, both in simulation and with real robots, in laboratory scenarios with two attacking robots versus two defending robots as well as versus a defender and a goalie. We show that we achieve the desired coordinated passing behavior, and also outperform a reactive action selection approach.     

Unstructured kinetic model for reuterin and 1,3‐propanediol production by Lactobacillus reuteri from glycerol/glucose cofermentation

BACKGROUND: Lactobacillus reuteri is unable to grow on glycerol as sole carbon and energy source hence, glycerol is used as an alternative hydrogen acceptor during growth on available carbohydrates. Thus, glycerol is converted to reuterin and 1,3‐propanediol (1,3‐PDL), both products with interesting industrial applications. These compounds are commonly produced by using resting cells in two‐step fermentation processes. RESULTS: The glycerol/glucose cofermentation by L. reuteri yields reuterin and 1,3‐PDL at a glycerol concentration higher than 100 mmol L⁻¹. An increase of glycerol concentration from 200 to 400 mmol L⁻¹ showed no additional stimulatory effect on ethanol and acetate production but consistently reduced the lactate concentration. It was also found that reuterin concentration reached a maximum value and subsequently decreased due to its conversion to 1,3‐PDL. An unstructured kinetic model was proposed to describe simultaneously microbial growth, substrates consumption and products formation. A multi‐response nonlinear regression analysis based on Marquardt algorithm combined with a Runge‐Kutta integration method was used to obtain the values of the fitting parameters. CONCLUSIONS: The optimum concentration of glycerol for maximum reuterin and 1,3‐PDL production was 200 mmol L⁻¹. The complete process was satisfactorily described by the kinetic model proposed. Copyright © 2008 Society of Chemical Industry     

Vinylic graft copolymers of cellulose. I. Effects of monomer concentrations and crystallinity index of cellulosic substrate on grafting vinyl acetate onto cotton

Graft copolymerization of vinyl acetate (VA) and methyl acrylate (MA) on cotton cellulose was initiated by the Ce (IV) ion, and ungrafted vinylic polymer was separated from the graft copolymer by acetone extraction. The influence of the ratio aqueous initiator solution volume/monomeric volume (V_aq/V_mon), vinyl acetate volume/methyl acrylate volume (V_VA/V_MA), and the cellulose crystallinity index (CI) on the grafting reaction were studied. To modify the crystallinity of cellulose, native cotton was treated with NaOH in the concentrations 10, 15, and 20% (mercerized). The viscosimetric average molecular weight (M_v), the polymerization degree (PD), and the crystallinity index proposed by Nelson and O'Connor (CI) were determined for native and NaOH-treated cotton. The polymeric side chains grafted were separated from the cellulose backbone by acid hydrolysis in 72% H₂SO₄. The viscosimetric average molecular weight (M_v) was determined, and the number of vinylic chains per cellulosic chain (graft frequency, GF) were calculated. The grafting percentage, %G, was higher for most amorphous cellulose and for a higher methyl acrylate percentage (%MA) in monomeric reaction mixtures (VA-MA). The V_aq/V_mon ratio that yields the highest %G was 70/30. The increase of the %G with the %MA in the VA–MA monomeric mixture seems to be due to both an increase in the length of vinylic grafted chains (as shown by its M_v) and the number of grafted chains (GF). The increase in the %G when the crystallinity index (CI) of the cellulosic substrate decreases seems to be due to an increase in the length of the vinylic grafted chains, but not to an increase in the number of grafted chains, since the M_v increases and GF decreases when the CI of cellulose decreases.     

Viscoelastic behaviour of vinylic copolymers of cellulose and their glass transition temperature by DSC

The viscoelastic response of some vinylic copolymers of cellulose prepared with vinyl acetatemethyl acrylate mixtures and with Ce(IV) ion as initiator, and native cellulose, were studied at 110 Hz in a range of temperatures from ?120–100°C. The viscoelastic spectrum of cellulose shows the β-relaxation that is not shown in its vinylic copolymers. We observed the same effect in the dielectric β-relaxation. For the vinylic copolymers of cellulose, one viscoelastic relaxation attributed to the α-relaxation of the grafted vinylic chains is observed. Some differences in the characteristics of this relaxation may be related to the composition of PVA/PMA vinylic side chains and to the ratio of cellulose in the copolymer. The plots of the Argand diagrams give us a better understanding of the viscoelastic behaviour of these materials. The results seem to indicate that the cellulose hinders the large-scale motions of the vinylic chains grafted onto it. The glass transition temperature (T_g) determined by differential scanning calorimetry (DSC) also shows the same fact: the T_g of the vinylic copolymers of cellulose are higher than both the T_g of polyvinyl acetate–polymethyl acrylate copolymers (PVA–PMA) without cellulose and the T_g of some blends of cellulose and the PVA–PMA whose composition was as similar as possible to the cellulosic copolymers. The importance of the covalent bonds between cellulose and the vinylic side chains in the structural transitions are revealed. The present results are compared with the dielectric α-relaxation that we described elsewhere. © 1993 John Wiley & Sons, Inc.     

Response surface optimisation of germination conditions to improve the accumulation of bioactive compounds and the antioxidant activity in quinoa

Germination has been proposed as an economic approach to improve the content of bioactive compounds in pseudocereals. In this work, response surface methodology (RSM) was employed to investigate the impact of germination conditions on the phytochemical content and antioxidant activity of quinoa. The use of desirability methodology showed that the optimum conditions to maximise the content of total phenolic content (TPC) and antioxidant activity in sprouted quinoa were 20 °C for 42 h. Sprouts produced under these conditions exhibited increases of 80% and 30% in TPC and antioxidant activity, respectively, compared to un‐germinated seeds, and contained high γ‐aminobutyric acid (GABA) concentration. The nonsignificant lack‐of‐fit and high determination coefficients obtained confirmed the suitability of the predictive models developed for TPC and antioxidant activity, whilst the one obtained for GABA was not significant (R² < 0.75) within the conditions studied. Sprouting under optimum conditions enhanced the content of both flavonoid and nonflavonoid compounds, being the increase in flavonoids more pronounced. Kaempferol‐O‐dirhamnosyl‐galactopyranose and quercetin‐O‐glucuronide were the compounds that experienced the most noticeable increase in quinoa after germination. In conclusion, this study provides useful information on the optimum germination conditions to improve the levels of health‐promoting compounds in quinoa.     

Computerised glow curve deconvolution using general and mixed order kinetics

Some accurate glow curve fitting functions for general and mixed order kinetics glow peaks are proposed and discussed. These mathematical expressions are used together with peak search and non-linear minimisation algorithms in order to provide a fast glow curve deconvolution for those materials which cannot be well fitted using first order kinetics. To test the accuracy of the proposed method. the result of the fitting of synthetic glow curves is compared with the original data giving negligible errors for values of parameters currently found in TL materials.     

Forthcoming Contributions

Table of Contents

Forthcoming Contributions     

Pitx2 participates in the late phase of the pathway controlling left-right asymmetry

BACKGROUND: We have studied the role of the different MHC (RT1) subregions in acute natural killer (NK) cell-mediated bone marrow allograft rejection in lethally irradiated, bone marrow cell (BMC) reconstituted rats. METHODS: We employed a series of MHC congenic and intra-MHC recombinant rat strains so that effects of mismatches in defined RT1 subregions could be studied systematically. BMC allograft survival was measured as 125IUdR uptake in the spleen between day 5 and day 7 after irradiation and BMC reconstitution. RESULTS: We found that in certain RT1 haplotype combinations, nonclassical RT1.C disparities by themselves could determine graft rejection (i.e., in the u/av1 recombinant haplotypes), whereas in another combination (between the av1 and c haplotypes) a mismatch for an isolated classical RT1.A region was decisive for engraftment. Thus, PVG.R1 BMC failed to proliferate in PVG rats, differing in the RT1.A region only, whereas in PVG.1U rats rejection could be determined by isolated differences in the RT1.C region (LEW.1WR1). Also, RT1 homozygous rats (RT1.U) rejected semi-allogeneic F1 hybrid BMC. The acute rejection of BMC was mediated by NK cells, as athymic nude rats, lacking alloreactive T cells but with normal alloreactive NK cells, showed the same patterns of rejection as did normal rats. Nude rats also rejected allogeneic lymphocytes, a previously documented NK-mediated phenomenon, with identical requirements of MHC disparity. CONCLUSIONS: This investigation shows that rat effector NK cells are radioresistant, independent of the thymus, and capable of recognizing and rejecting MHC mismatched transplanted BMC on the basis of mismatches in both classical and nonclassical class I regions in vivo. The studies underline the importance also of NK cells in determining BMC allograft survival.