The dynamic process of dietary soybean ß-conglycinin in digestion, absorption, and metabolism among different intestinal segments in grass carp (Ctenopharyngodon idella).

The present study aimed to investigate the dynamic process of soybean ß-conglycinin in digestion, absorption, and metabolism in the intestine of grass carp (Ctenopharyngodon idella). Fish fed with 80 g ß-conglycinin/kg diet for 7 weeks, the intestinal digestive enzyme was extracted to hydrolyze ß-conglycinin in vitro, the free amino acid and its metabolism product contents in intestinal segments were analyzed. The present study first found that ß-conglycinin cannot be thoroughly digested by fish intestine digestive enzyme and produces new products (about 60- and 55-kDa polypeptides). The indigestible ß-conglycinin further caused the free amino acid imbalance, especially caused free essential amino acid deficiency in the proximal intestine but excess in the distal intestine. Moreover, these results might be partly associated with the effect of ß-conglycinin in amino acid transporters and tight junction-regulated paracellular pathway. Finally, dietary ß-conglycinin increased the content of amino acid catabolism by-product ammonia while decreased the amino acid anabolism product carnosine content in the proximal intestine and distal intestine. Thus, the current study first and systemically explored the dynamic process of ß-conglycinin in digestion, absorption, and metabolism, which further supported our previous study that dietary ß-conglycinin suppressed fish growth and caused intestine injure.

Identification of Triticum aestivum nsLTPs and functional validation of two members in development and stress mitigation roles.

Role of plant nsLTP in biotic stress is well reported; however, their role during abiotic stress is far from clear. This study comprises genome-wide identification of LTPs and characterizes the regulation and function of two Triticum aestivum lipid transfer proteins, TaLTP40 and TaLTP75, under stresses that influence membrane fluidity. A total of 105 LTP gene family members have been identified. The selected LTPs for functional validation were highly expressed during salt, cold and drought stress. Further, selected LTPs showed differential expression thermotolerant and thermosusceptible wheat cultivars. Higher expression of many TaLTPs was observed under different abiotic stresses in thermotolerant wheat cultivars as compared to thermosusceptible cultivars. TaLTPs regulation was correlated with light energy distribution studies under similar stress conditions. Cellular localization revealed localization of different TaLTPs to the tonoplast membrane along with the organelles involved in the secretory pathway. Induction of TaLTPs was observed upon treatment with dimethylsulphoxide. TaLTP40 and TaLTP75 overexpressing transgenic Arabidopsis showed a constitutively enhanced salt tolerance. Both the TaLTP40 and TaLTP75 overexpressing lines performed better in terms of chlorophyll a fluorescence, total chlorophyll content, membrane injury index, total biomass, percentage germination, percentage survival and relative growth rate. Hence, our analyses indicate that TaLTPs expression might be driven by change in membrane fluidity and could be involved in transferring membrane lipids to the biological membranes thus imparting tolerance to various abiotic stresses.

GhLTPG1, a cotton GPI-anchored lipid transfer protein, regulates the transport of phosphatidylinositol monophosphates and cotton fiber elongation.

The cotton fibers are seed trichomes that elongate from the ovule epidermis. Polar lipids are required for the quick enlargement of cell membrane and fiber cell growth, however, how lipids are transported from the ovules into the developing fibers remains less known. Here, we reported the functional characterization of GhLTPG1, a GPI-anchored lipid transport protein, during cotton fiber elongation. GhLTPG1 was abundantly expressed in elongating cotton fibers and outer integument of the ovules, and GhLTPG1 protein was located on cell membrane. Biochemical analysis showed that GhLTPG1 specifically bound to phosphatidylinositol mono-phosphates (PtdIns3P, PtdIns4P and PtdIns5P) in vitro and transported PtdInsPs from the synthesis places to the plasma membranes in vivo. Expression of GhLTPG1 in Arabidopsis caused an increased number of trichomes, and fibers in GhLTPG1-knockdown cotton plants exhibited significantly reduced length, decreased polar lipid content, and repression of fiber elongation-related genes expression. These results suggested that GhLTPG1 protein regulates the cotton fiber elongation through mediating the transport of phosphatidylinositol monophosphates.

Elusive Structural, Functional, and Immunological Features of Act d 5, the Green Kiwifruit Kiwellin.

Kiwellin (Act d 5) is an allergenic protein contained in kiwifruit pulp in high amounts. The aim of this study was to investigate the three-dimensional structure of the natural molecule from green kiwifruit and its possible function. Kiwellin was crystallized, and its structure, including post-translational modifications, was elucidated. The molecular weight and structural features, in solution, were analyzed by gel filtration and circular dichroism, respectively. Although structurally similar to expansin, kiwellin lacks expansin activity and carbohydrate binding. A specific algorithm was applied to investigate any possible IgE reactivity correlation between kiwellin and a panel of 102 allergens, including expansins and other carbohydrate-binding allergens. The available data suggest a strong dependence of the kiwellin structure on the environmental/experimental conditions. This dependence therefore poses challenges in detecting the correlations between structural, functional, and immunological features of this protein.

Motivation enhances visual working memory capacity through the modulation of central cognitive processes.

Motivation is well known to enhance working memory (WM) capacity, but the mechanism underlying this effect remains unclear. The WM process can be divided into encoding, maintenance, and retrieval, and in a change detection visual WM paradigm, the encoding and retrieval processes can be subdivided into perceptual and central processing. To clarify which of these segments are most influenced by motivation, we measured ERPs in a change detection task with differential monetary rewards. The results showed that the enhancement of WM capacity under high motivation was accompanied by modulations of late central components but not those reflecting attentional control on perceptual inputs across all stages of WM. We conclude that the "state-dependent" shift of motivation impacted the central, rather than the perceptual functions in order to achieve better behavioral performances.

The involvement of Medicago truncatula non-specific lipid transfer protein N5 in the control of rhizobial infection.

Cysteine-rich proteins seem to play important regulatory roles in Medicago truncatula/Sinorhizobium meliloti symbiosis. In particular, a large family of nodule-specific cysteine-rich (NCR) peptides is crucial for the differentiation of nitrogen-fixing bacteroids. The Medicago truncatula N5 protein (MtN5) is currently the only reported non-specific lipid transfer protein necessary for successful rhizobial symbiosis; in addition, MtN5 shares several characteristics with NCR peptides: a small size, a conserved cysteine-rich motif, an N-terminal signal peptide for secretion and antimicrobial activity. Unlike NCR peptides, MtN5 expression is not restricted to the root nodules and is induced during the early phases of symbiosis in root hairs and nodule primordia. Recently, MtN5 was determined to be involved in the regulation of root tissue invasion; while, it was dispensable for nodule primordia formation. Here, we discuss the hypothesis that MtN5 participates in linking the progression of bacterial invasion with restricting the competence of root hairs for infection.

The rice OsDIL gene plays a role in drought tolerance at vegetative and reproductive stages.

Drought is one of the critical factors limiting reproductive yields of rice and other crops globally. However, little is known about the molecular mechanism underlying reproductive development under drought stress in rice. To explore the potential gene function for improving rice reproductive development under drought, a drought induced gene, Oryza sativa Drought-Induced LTP (OsDIL) encoding a lipid transfer protein, was identified from our microarray data and selected for further study. OsDIL was primarily expressed in the anther and mainly responsive to abiotic stresses, including drought, cold, NaCl, and stress-related plant hormone abscisic acid (ABA). Compared with wild type, the OsDIL-overexpressing transgenic rice plants were more tolerant to drought stress during vegetative development and showed less severe tapetal defects and fewer defective anther sacs when treated with drought at the reproductive stage. The expression levels of the drought-responsive genes RD22, SODA1, bZIP46 and POD, as well as the ABA synthetic gene ZEP1 were up-regulated in the OsDIL-overexpression lines but the ABA degradation gene ABAOX3 was down-regulated. Moreover, overexpression of OsDIL lessened the down-regulation by drought of anther developmental genes (OsC4, CYP704B2 and OsCP1), providing a mechanism supporting pollen fertility under drought. Overexpression of OsDIL significantly enhanced drought resistance in transgenic rice during reproductive development, while showing no phenotypic changes or yield penalty under normal conditions. Therefore, OsDIL is an excellent candidate gene for genetic improvement of crop yield in adaption to unfavorable environments.

Directed forgetting in direct and indirect tests of memory: seeking evidence of retrieval inhibition using electrophysiological measures.

We investigated whether directed forgetting as elicited by the item-cueing method results solely from differential rehearsal of to-be-remembered vs. to-be-forgotten words or, additionally, from inhibitory processes that actively impair retrieval of to-be-forgotten words. During study, participants (N=24) were instructed to remember half of a series of presented words (TBR) and to forget the other half (TBF), as indicated by an instruction cue shown shortly after each word. During test, accuracy and reaction time measures from lexical decisions (indirect memory test) followed by recognition-memory judgements (direct memory test) were supplemented with event-related potential (ERP) recordings. Results from the behavioural measures revealed directed forgetting in the recognition-memory test but not the lexical-decision test. ERPs obtained during recognition indicated that TBR words elicited a larger parietal old/new effect than TBF words overall, suggesting that remember/forget instructions impaired conscious recollection processes more severely than familiarity processes. Moreover, TBF words that were successfully forgotten elicited less parietal activity than correctly rejected new words (the reversed old/new effect; Nowicka, A., Jednorórog, K., Wypych, M., & Marchewka, A. (2009). Reversed old/new effect for intentionally forgotten words: An ERP study of directed forgetting. International Journal of Psychophysiology, 71, 97-102). This was taken to implicate that inhibitory processes likely affected these items. Enhanced negativities for successfully forgotten TBF words relative to new words were observed in the lexical-decision task at early (150-250 ms) and late (800-1000 ms) time windows, suggesting that inhibitory influences disrupt more than just conscious recollection when memory retrieval is tested indirectly.

Caveolar transport through nasal epithelium of birch pollen allergen Bet v 1 in allergic patients.

BACKGROUND: Previous work in type I pollen allergies has focused on aberrant immunoresponses. OBJECTIVE: Our systems-level analyses explore the role of epithelium in early pathogenesis of type I allergic reactions. METHODS: We began top-down analyses of differences in human nasal epithelial cells and biopsy specimens obtained from patients with birch allergy and healthy control subjects in the resting state and after intranasal in vivo birch pollen challenges. Immunohistochemistry, immunotransmission electron microscopy, mass spectrometry, transcriptomics, and integration of data to a pathway were conducted. RESULTS: Bet v 1 allergen bound to epithelium immediately after in vivo birch pollen challenge during winter only in allergic individuals. It also travelled through epithelium with caveolae to mast cells. Sixteen unique proteins were found to bind to the Bet v 1 column only in lysates from allergic epithelial cells; 6 of these were caveolar and 6 were cytoskeletal proteins. The nasal epithelial transcriptome analysis from allergic and healthy subjects differed during the winter season, and these subjects also responded differentially to birch pollen challenge. Within this pollen-induced response, the gene ontology categories of cytoskeleton and actin cytoskeleton were decreased in allergic patients, whereas the actin-binding category was enriched in healthy subjects. Integration of microscopic, mass spectrometric, and transcriptomic data to a common protein-protein binding network showed how these were connected to each other. CONCLUSION: We propose a hypothesis of caveolae-dependent uptake and transport of birch pollen allergen in the epithelium of allergic patients only. Application of discovery-driven methodologies can provide new hypotheses worth further analysis of complex multifactorial diseases, such as type I allergy.

The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs).

Plant lipid-transfer proteins (LTPs) are abundant, small, lipid binding proteins that are capable of exchanging lipids between membranes in vitro. Despite their name, a role in intracellular lipid transport is considered unlikely, based on their extracellular localization. A number of other biological roles, including antimicrobial defense, signaling, and cell wall loosening, have been proposed, but conclusive evidence is generally lacking, and these functions are not well correlated with in vitro activity or structure. A survey of sequenced plant genomes suggests that the two biochemically characterized families of LTPs are phylogenetically restricted to seed plants and are present as substantial gene families. This review aims to summarize the current understanding of LTP biochemistry, as well as the evidence supporting the proposed in vivo roles of these proteins within the emerging post-genomic framework.

Lipid transfer protein allergy: primary food allergy or pollen/food syndrome in some cases.

PURPOSE OF REVIEW: To summarize recent findings on non-specific lipid transfer proteins in food allergy, with a specific focus on the localization, stability and route of sensitization. RECENT FINDINGS: Plant non-specific lipid transfer proteins are major food allergens, especially in the Mediterranean area. They have been identified as allergens in a number of foods and the list grows ever longer. As non-specific lipid transfer proteins are considered to be "true" food allergens that sensitize directly via the gastrointestinal tract their stability during food processing and gastric digestion has been studied in more detail. In addition, several groups have tried to determine the sensitization patterns of lipid transfer protein-reactive patients, to determine and possibly clarify the observed geographical differences in sensitization. Different sensitization routes (via the respiratory tract or even transdermally) have been suggested. SUMMARY: As the structure and molecular properties of non-specific lipid transfer proteins are resolved and more purified non-specific lipid transfer proteins become available for diagnostic purposes, detailed studies on the sensitization pattern and route are becoming feasible. Continuing studies on the pattern of lipid transfer protein sensitization will give more insight into the development and possible treatment of lipid transfer protein-related food allergy.

Role of plant lipid transfer proteins in plant cell physiology-a concise review.

Plant lipid transfer proteins (LTP) are cationic peptides, subdivided into two families, which present molecular masses of around 7 and 10 kDa. The peptides were, thus, denominated due to their ability to reversibly bind and transport hydrophobic molecules in vitro. Both subfamilies possess conserved patterns of eight cysteine residues and the three-dimensional structure reveals an internal hydrophobic cavity that comprises the lipid binding site. Based on the growing knowledge regarding structure, gene expression and regulation and in vitro activity, LTPs are likely to play a role in key processes of plant physiology. Although the roles of plant LTPs have not yet been fully determined. This review aims to present comprehensive information of recent topics, cover new additional data, and present new perspectives on these families of peptides.