Search

Six pairs of near isogenic wheat lines with and without the 1BL.1RS wheat-rye chromosome translocation originating from Austria and Hungary were grown in eleven environments and analysed for agronomic and quality traits. Stability across environments, estimated by ecovalence and mean deviation from regression, was calculated for yield, test weight, protein and wet gluten content, and SDS-protein gel test. Yield stability parameters were found to be similar for both translocated and non-translocated lines, whereas for the other investigated traits clear differences were found. Higher stability, however, could not generally be attributed to translocated or non-translocated lines. Thus, for environmental stability the genotype of the wheat background appears to be more important than the presence of the translocation. Combining the means for yield, test weight, protein and wet gluten content and their environmental stabilities revealed four genotypes performing better than average. These included two genotypes carrying the 1BL.1RS translocation and two without the translocation. However, when gluten quality is additionally considered, as measured by the SDS-protein gel test, the two translocation lines fall back below average. These observations and the results of a literature survey led us to conclude, that the value of the 1BL.1RS translocation for wheat breeding in its present form is indeed questionable, especially if the often associated unfavourable effects of 1RS on gluten and bread making quality are considered. [Copyright &y& Elsevier]

Isoprenoids are synthesised either through the classical, mevalonate pathway, or the alternative, non-mevalonate, 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. The latter is found in many microbial pathogens and proceeds via (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP), a potent activator of human Vγ9/Vδ2 T cells. Listeria monocytogenes is the only pathogenic bacterium known to contain both pathways concurrently. Strategic gene knockouts demonstrate that either pathway is functional but dispensable for viability. Yet, disrupting the mevalonate pathway results in a complementary upregulation of the MEP pathway. Vγ9/Vδ2 T cell bioactivity is increased in ΔlytB mutants where HMB-PP accumulation is expected, and lost in ΔgcpE mutants which fail to produce HMB-PP. [Copyright &y& Elsevier]

The extension testing of wheat-flour dough has become one of the key cereal chemistry links to end product quality assessment, because of its perceived relevance to baking performance, and because of the various correlations that have been inferred and assumed since the extensibility of a dough was first investigated in the late 19th century. As a consequence, from a plant breeding perspective, there is a need to understand and interpret the extensibility of a dough in terms of the molecular dynamics occurring during its extension, not only as it relates to its baking, but also to the HMW/LMW glutenin and gliadin composition of the wheat. The focus of this paper is the direct measurement of extensibility on a micro-extension tester. The theoretical justification is that the resulting extensogram represents an encapsulation of the extensional rheology of the dough. The paper reports on a graphical and statistical analysis of extension tests performed on eight representative flours. They yield validation for the conclusion that the historic assessment of extensibility as ExtRupture should be replaced by ExtRmax and the difference ExtRupture−ExtRmax. It is relatively easy to obtain accurate estimates of Rmax, ExtRmax and ExtRupture from electronically recorded extensograms. Among other things, it is established that, in terms of the glutenin classification of flours based on electrophoresis, HPLC or DNA markers, Rmax yields a far better differentiation than is achieved with either ExtRupture or ExtRmax. It follows, as a corollary to such results, that the extensions to the earlier rheological phases in an extensogram are just as, if not more, important in identifying connections between the gluten composition of wheat varieties and the traditional ExtRupture and Rmax. In particular, it is shown that, like Rmax, the scaled difference Rmax/(ExtRupture−ExtRmax) represents an alternative strategy to relate quality to the glutenin composition of wheat varieties, determined by electrophoresis, HPLC and DNA markers. [Copyright &y& Elsevier]

We have used two fluorescent probes, NBD and dansyl, attached site-specifically to the serpin plasminogen activator inhibitor-1 (PAI-1) to address the question of whether a common mechanism of proteinase translocation and full insertion of the reactive center loop is used by PAI-1 when it forms covalent SDS-stable complexes with four arginine-specific proteinases, which differ markedly in size and domain composition. Single-cysteine residues were incorporated at position 119 or 302 as sites for specific reporter labeling. These are positions ∼30 Å apart that allow discrimination between different types of complex structure. Fluorescent derivatives were prepared for each of these variants using both NBD and dansyl as reporters of local perturbations. Spectra of native and cleaved forms also allowed discrimination between direct proteinase-induced changes and effects solely due to conformational change within the serpin. Covalent complexes of these derivatized PAI-1 species were made with the proteinases trypsin, LMW u-PA, HMW u-PA, and t-PA. Whereas only minor perturbations of either NBD and dansyl were found for almost all complexes when label was at position 119, major perturbations in both wavelength maximum (blue shifts) and quantum yield (both increases and decreases) were found for all complexes for both NBD and dansyl at position 302. This is consistent with all four complexes having similar location of the proteinase catalytic domain and hence with all four using the same mechanism of full-loop insertion with consequent distortion of the proteinase wedged in at the bottom of the serpin. [ABSTRACT FROM AUTHOR]

Background/aims: GAPD has been exhaustively investigated as a key cytosolic enzyme in glycolysis. In recent years GAPD has also been implicated in many cellular activities unrelated to glycolysis. However, although various functions have been ascribed to GAPD from rabbit muscle, human blood and rat tissues, no information is available on human liver GAPD. We have recently demonstrated that, as a cellular kinase, GAPD might interfere in the life-cycle of hepatitis B virus. We therefore investigated the enzymatic activities and subcellular localization of GAPD in normal human liver. Methods: GAPD and hepatocyte membranes were isolated from human liver homogenates to study the subcellular localization and enzymatic activities of GADP (kinase and ADP-ribosyltransferase). Results: (i) GAPD was recovered from the plasma-membrane-enriched fraction, in internal membranes, and in the cytosol; (ii) GAPD could be phosphorylated, a phenomenon inhibited by both GAP and NADH; and (iii) GAPD exhibits ADP-ribosyltransferase activity, which is stimulated by nitric oxide in a concentration-dependent manner. Conclusions: Human liver GAPD may play significant biological roles in addition to glycolysis, especially in signal transduction and in intracellular processes possibly involved in HBV infection. [ABSTRACT FROM AUTHOR]

The effects of various purification techniques on kiwifruit enzyme characteristics (protease activity, kinetic parameters, and protein patterns) and production of wheat gluten bio-active peptides were investigated. The enzyme extract purified by ammonium sulfate precipitation method exhibited the highest protease activity (26), K m (0.04 ± 0.002 mM), K cat / K m (40), and yield (96%). Using actinidin, the hard and soft wheat gluten subunit proteins produced antidiabetic inhibitory (α-glucosidase and α-amylase) peptides. The smallest M w fraction of soft wheat gliadin peptide (<1 kDa) showed the highest inhibitory capacity against α-glucosidase (18.4 ± 0.7%) and α-amylase (53.3 ± 1.9%). The presence of high levels of amino acids with hydroxyl groups and proline in P3 sub-fraction had a critical role on α-glucosidase (47.2%) and α-amylase (71.2%) inhibitory activities. In conclusion, wheat gluten subunit peptides showed significant metabolic effects relevant to glucose and insulin control in vitro., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Marine ecosystems are highly dependent on macroalgea in providing food and shelter for aquatic organisms, interacting with many bacteria and mostly producing secondary metabolites of potent therapeutic antibacterial property. Screening of marine microbial secondary metabolites of valuable biotechnological and therapeutical applications are now extensively studied. In this study, Bacillus spp. identified by DNA sequencing and found associated with Turbinaria ornata , was screened and characterized for its cell free supernatant (CFS) possible antimicrobial and antibiofilm applications. Among the 7 microbial isolates tested, CFS greatly affected Bacillus subitilis (12 mm) and inhibited equally the yeast isolates Candida albicans , Candida tropicalis and Candida glabrata (10 mm) and had no or negligible effect on S.aureus, E.coli, P. aeruginosa. As for the CFS antibiofilm activity, no difference was revealed from the positive control. Algal crude extracts (methanol, acetone and aqueous), on the other hand, were similarly tested for their antimicrobial activity against the seven microbial isolates, where highest activity was observed with the aqueous crude extract against Staphylococcus aureus (10 mm) and Pseudomonas aeruginosa (9 mm) compared to the negligible effects of methanol and acetone crude extracts. Chemical analysis was performed to reveal the major constituents of both crude algal extracts and Bacillus spp. CFS. FTIR spectrum of the bacterial CFS indicated the presence of bacteriocin as the major lipopeptide responsible for its biological activity. Whereas, methanol and water crude algal extract GC-MS spectra revealed different chemical groups of various combined therapeutical activity mainly Naphthalene, amino ethane-sulfonic acid, pyrlene, Biotin and mercury chloromethyl correspondingly. Thus, the present study, demonstrated the moderate activity of both crude algal extract and the bacterial CFS, however, further investigations are needed for a better biological activity., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

The starchy endosperm of the mature wheat grain comprises three major cell types, namely sub-aleurone cells, prismatic cells and central cells, which differ in their contents of functional components: gluten proteins, starch, cell wall polysaccharides (dietary fibre) and lipids. Gradients are established during grain development but may be modified during grain maturation and are affected by plant nutrition, particularly nitrogen application, and environmental factors. Although the molecular controls of their formation are unknown, the high content of protein and low content of starch of sub-aleurone cells, compared to the other starchy endosperm cells types, may result from differences in developmental programming related to the cells having a separate origin (from anticlinal division of the aleurone cells). The gradients within the grain may be reflected in differences in the compositions of mill streams, particularly those streams enriched in the central and outer cells of the starchy endosperm, respectively, allowing the production of specialist flours for specific end uses., (© 2019 The Authors.)

Objectives: Low-molecular-weight (LMW) substances are known to be causative agents of occupational asthma (OA) and occupational rhinitis (OR). Although most LMW substances are irritants or allergens, some can cause immediate type immunoglobulin E (IgE)-mediated allergic reactions. Trimellitic anhydride (TMA) is one such LMW substance, which is known as an immunological sensitizer. However, the exact molecular biological details of the effects of TMA remain unclear., Methods: We measured the β-hexosaminidase release from mast cells after directly exposing the cells to various LMW substances. The tyrosine phosphorylation of whole cellular molecules and the phosphorylation of extracellular signal-regulated kinase (ERK) were assessed by immunoblot assay., Results: Among the LMW substances tested, only TMA induced β-hexosaminidase release. However, the mast cell degranulation induced by TMA was lower than that induced by an antigen or a calcium ionophore. Moreover, the pattern of tyrosine phosphorylation of whole cellular molecules was quite different between IgE-mediated antigen stimulation and TMA exposure. The TMA effect on mast cells was independent of not only IgE but also Ca 2+ influx. ERK phosphorylation was not detected in mast cells exposed to TMA., Conclusions: TMA induced mild degranulation of mast cells without IgE, even though the phosphorylation of ERK was not detected. This reaction suggests that TMA affects humans even upon first exposure. Therefore, it is imperative to avoid human exposure to high concentrations of TMA. In order to stop the development of severe asthma in individuals with OR, we need to be able to identify cases of OR caused by TMA as soon as possible.

Whey-acidic-protein (WAP) four-disulphide core (WFDC) proteins have important roles in the regulation of innate immunity, anti-microbial function, and the inhibition of inflammatory proteases at mucosal surfaces. It was recently demonstrated that the WFDC protein, prostate stromal 20 (ps20), encoded by the WFDC1 gene, is a potent growth inhibitory factor, and shares with other WFDC proteins the ability to modulate wound healing processes and immune responses to viral infections. However, ps20 remains relatively uncharacterised at the protein level. Using a panel of ps20 antibodies for western-blotting (WB), ELISA and immunoaffinity purification, we isolated, biochemically characterised and tested ps20 preparations for three biological properties: (i) interactions with glycosaminoglycans (GAG) (ii) inhibition of cell proliferation, and (iii) transglutaminase2 (TG2) mediated crosslinking of ps20 to fibronectin, a process implicated in wound healing. We show herein that ps20 preparations contain multiple molecular forms including full-length ps20 (resolving at ≈27 kDa), an exon 3 truncated form (≈22 kDa) that lacks aa113-140, and variable amounts of a putatively cleaved lower MW (≈15-17 kDa) species. Untagged purified ps20 preparations containing a mixture of these forms are biologically active in significantly suppressing prostate cell proliferation. We show that one mechanism by which lower LMW forms of ps20 arise is through cathepsin L (CL) cleavage, and confirm that CL cleaves ps20 at the C-terminus, but this does not inhibit its growth inhibitory function. However, CL cleavage abrogated the interaction between ps20 and solid-phase fibronectin. Therefore, we demonstrate for the first time that LMW forms of ps20 that lack a C-terminal immunogenic epitope can arise through CL cleavage and this cleavage impairs multimerisation and potential capacity to cross-link to ECM, but not the capacity of ps20 to inhibit cell proliferation. We propose that ps20 like other WFDC proteins can become associated with GAGs and the ECM. Furthermore, we suggest post-translational processing and cleavage of ps20 is required to generate functional protein species, and TG2 mediated crosslinking and CL cleavage form components of a ps20 regulatory apparatus.

Objective: Plasma total adiponectin reveals a sexual dimorphism indicating that gonadal steroids may be involved in its secretion and/or metabolism. However, results from previous reports are conflicting and data regarding the influence of ovarian steroids on adiponectin's multimer forms are scarce. The objective of the study was to assess if total adiponectin and its isoforms are affected by the changes of estradiol and progesterone during the normal menstrual cycle and the association of total adiponectin and its isoforms with the gonadal steroid levels., Materials/methods: Quantitative determination of plasma adiponectin and its multimers was conducted in the three phases of an ovulatory cycle in 13 premenopausal women, in the follicular phase of 10 more premenopausal women, in 20 postmenopausal women and in 21 men. Moreover, serum levels of FSH, LH, prolactin, estradiol, progesterone, and testosterone, sex hormone binding globulin, glucose, and insulin were measured., Results: The circulating levels of total adiponectin and its multimers were not affected by the normal variation of estradiol and progesterone across the ovulatory menstrual cycle. In the whole number of participants, the total adiponectin and high molecular weight adiponectin levels were significantly different between genders and associated positively with age and sex hormone binding globulin levels, and negatively with testosterone and progesterone levels and the waist/hip ratio. In the multiple logistic regression analysis, after adjustment for age, gender, and sex hormone binding globulin and progesterone levels, significant predictors of total adiponectin levels were the waist/hip ratio and testosterone levels, and of high molecular weight adiponectin the testosterone levels., Conclusions: Normal menstrual cycle ovarian steroids are not involved directly in the regulation of secretion and/or metabolism of total adiponectin and its multimers. Testosterone seems to be responsible for the adiponectin's sexual dimorphism.

The worldwide emergence of epidemic strains of Clostridium difficile linked to increased disease severity and mortality has resulted in greater research efforts toward determining the virulence factors and pathogenesis mechanisms used by this organism to cause disease. C. difficile is an opportunist pathogen that employs many factors to infect and damage the host, often with devastating consequences. This review will focus on the role of the 2 major virulence factors, toxin A (TcdA) and toxin B (TcdB), as well as the role of other putative virulence factors, such as binary toxin, in C. difficile-mediated infection. Consideration is given to the importance of spores in both the initiation of disease and disease recurrence and also to the role that surface proteins play in host interactions.