Reg-1α, a New Substrate of Calpain-2 Depending on Its Glycosylation Status

Reg-1α/lithostathine, a protein mainly associated with the digestive system, was previously shown to be overexpressed in the pre-clinical stages of Alzheimer’s disease. In vitro, the glycosylated protein was reported to form fibrils at physiological pH following the proteolytic action of trypsin. However, the nature of the protease able to act in the central nervous system is unknown. In the present study, we showed that Reg-1α can be cleaved in vitro by calpain-2, the calcium activated neutral protease, overexpressed in neurodegenerative diseases. Using chemical crosslinking experiments, we found that the two proteins can interact with each other. Identification of the cleavage site using mass spectrometry, between Gln⁴ and Thr⁵, was found in agreement with the in silico prediction of the calpain cleavage site, in a position different from the one reported for trypsin, i.e., Arg¹¹-Ile¹² peptide bond. We showed that the cleavage was impeded by the presence of the neighboring glycosylation of Thr⁵. Moreover, in vitro studies using electron microscopy showed that calpain-cleaved protein does not form fibrils as observed after trypsin cleavage. Collectively, our results show that calpain-2 cleaves Reg-1α in vitro, and that this action is not associated with fibril formation.     

Studies on curcumin and curcuminoids XXI. Variation in the content of curcuminoids in Curcuma longa L. and Curcuma aromatica Salisb. from India during one season

Summary Three varieties ofCurcuma longa andC. aromatica were investigated during a growth period of 17 weeks with respect to the content of curcuminoids in the bulbs and fingers. A decrease in pigment production as a function of maturity of the rhizomes was observed. The total content of curcuminoids was equal inC. longa andC. aromatica, but with a slight difference in distribution between the fingers and bulbs in the two species. Taking the biomass into account, the fingers will be more beneficial for the isolation of curcuminoids than the respective bulbs.

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Studien über Curcumin und Curcuminoide XXI. Unterschiede des Gehaltes an Curcuminoiden bei Curcuma longa L. und Curcuma aromatica Salisb. aus Indien während einer Saison

Zusammenfassung Drei Typen vonCurcuma longa undC. aromatica wurden während einer Wachstumsperiode von 17 Wochen auf ihren Curcuminoidgehalt in den Knollen und im Wurzelstock untersucht. Eine Abnahme der Pigmentproduktion in Abhängigkeit von dem Entwicklungsstadium der Rhizome wurde festgestellt. Der Gesamtgehalt an Curcuminoiden beiC. longa undC. aromatica war der gleiche, doch bestand ein geringer Unterschied in der Verteilung dieser zwischen Knolle und Wurzelstock bei den beiden Spezies. Bei Betrachtung der Biomasse ist der Wurzelstock für die Isolation von Curcuminoiden besser geeignet als die Knollen.

     

Echium oil is better than rapeseed oil in improving the response of barramundi to a disease challenge

Pathogen infection stimulates the fatty acid (FA) metabolism and the production of pro-inflammatory derivatives of FA. Barramundi, Lates calcarifer, was fed on a diet rich in preformed long-chain (?C₂₀) polyunsaturated fatty acids (LC-PUFA) from fish oil (FO), to compare with diets containing high levels of C₁₈ precursors for LC-PUFA – stearidonic (SDA) and γ-linolenic acid (GLA) – from Echium plantagineum (EO), or rapeseed oil (RO) rich in α-linolenic acid (ALA), but a poor source of LC-PUFA and their precursors. After 6 weeks, when growth rates were similar amongst the dietary treatments, a sub-lethal dose of Streptococcus iniae was administered to half of the fish, while the other half were maintained unchallenged and were pair-fed with the infected fish. Under a disease challenge situation, the tissue FA depots depleted at 3 days post-infection (DPI) and were then restored to their previous concentrations at 7 DPI. During the infection period, EO fish had a higher content of n3 and n6 PUFA in their tissues, higher n3:n6 PUFA ratio and reduced levels of the eicosanoids, TXB₂ and 6-keto-PGF_1α, in their plasma compared with RO fish. Fish fed on FO and EO had a longer lasting and enduring response in their FA and eicosanoid concentrations, following a week of bacterial infection, compared with those fed on RO. EO, containing SDA and GLA and with a comparatively higher n3:n6 PUFA ratio, proved more effective than RO in compensating for immunity stress.     

Design space exploration for real-time embedded stream processors

We explore tradeoffs between organization and number of ALUs and clock frequency in a stream processor. The tool provides candidate low-power configurations and estimates of their real-time performance. The tool relates instruction-level, subword, and data parallelism to functional units' organization and utilization. The exploration methodology is applicable to all embedded-processor designs in signal and media processing.     

Non-targeted screening for contaminants in paper and board food-contact materials using effect-directed analysis and accurate mass spectrometry

Due to large knowledge gaps in chemical composition and toxicological data for substances involved, paper and board food-contact materials (P&B FCM) have been emerging as a FCM type of particular concern for consumer safety. This study describes the development of a step-by-step strategy, including extraction, high-performance liquid chromatography (HPLC) fractionation, tentative identification of relevant substances and in vitro testing of selected tentatively identified substances. As a case study, we used two fractions from a recycled pizza box sample which exhibited aryl hydrocarbon receptor (AhR) activity. These fractions were analysed by gas chromatography (GC) and ultra-HPLC (UHPLC) coupled to quadrupole time-of-flight mass spectrometers (QTOF MS) in order tentatively to identify substances. The elemental composition was determined for peaks above a threshold, and compared with entries in a commercial mass spectral library for GC-MS (GC-EI-QTOF MS) analysis and an in-house built library of accurate masses for substances known to be used in P&B packaging for UHPLC-QTOF analysis. Of 75 tentatively identified substances, 15 were initially selected for further testing in vitro; however, only seven were commercially available and subsequently tested in vitro and quantified. Of these seven, the identities of three pigments found in printing inks were confirmed by UHPLC tandem mass spectrometry (QqQ MS/MS). Two pigments had entries in the database, meaning that a material relevant accurate mass database can provide a fast tentative identification. Pure standards of the seven tentatively identified substances were tested in vitro but could not explain a significant proportion of the AhR-response in the extract. Targeted analyses of dioxins and PCBs, both well-known AhR agonists, was performed. However, the dioxins could explain approximately 3% of the activity observed in the pizza box extract indicating that some very AhR active substance(s) still remain to be identified in recycled low quality P&B.     

The effect of breed,parity, and stage of lactation on conjugated linoleic acid (CLA) in milk fat from dairy cows

Dairy products are the main source of conjugated linoleic acid (CLA), a functional food component with health benefits. The major source of cis-9, trans-11 CLA in milk fat is endogenous synthesis via delta9-desaturase from trans-11 18:1, with the remainder from incomplete rumen biohydrogenation of linoleic acid. Diet has a major influence on milk fat CLA; however, effects of physiological factors have received little attention. Our objectives were to examine milk fat content of CLA and the CLA-desaturase index with regard to: 1) effect of breed, parity, and stage of lactation, and 2) variation among individuals and the relationship to milk and milk fat. Holstein (n = 113) and Brown Swiss (n = 106) cows were fed a single diet and milk sampled on the same day to avoid confounding effects of diet and season. Frequency distributions demonstrated that milk fat content of CLA and CLA-desaturase index varied over threefold among individuals, and this needs to be considered in the design of experiments. Holsteins had a higher milk fat content of CLA and CLA-desaturase index, but breed differences were minor. Parity and days in milk also had little or no relationship to the individual variation for these two CLA variables. Breed, parity, and days in milk accounted for < 0.1, < 0.3, and < 2.0% of total variation in CLA concentration in milk fat, respectively. Milk fat content of CLA and CLA-desaturase index were essentially independent of milk yield, milk fat percent, and milk fat yield. We speculate that the basis for the genetic variation among individuals is related to rumen output of trans-11 18:1 and to a lesser extent cis-9, trans-11 CLA, and to the tissue amount and activity of delta9-desaturase.