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Adhesive-invasive E. coli has been suggested to be associated with the development of Crohn's disease (CD). It is assumed that they can provoke the onset of the inflammatory process as a result of the invasion of intestinal epithelial cells and then, due to survival inside macrophages and dendritic cells, stimulate chronic inflammation. In previous reports, we have shown that passage of the CD isolate ZvL2 on minimal medium M9 supplemented with sodium propionate (PA) as a carbon source stimulates and inhibits the adherent-invasive properties and the ability to survive in macrophages. This effect was reversible and not observed for the laboratory strain K12 MG1655. We were able to compare the isogenic strain AIEC in two phenotypes—virulent (ZvL2-PA) and non-virulent (ZvL2-GLU). Unlike ZvL2-GLU, ZvL2-PA activates the production of ROS and cytokines when interacting with neutrophils. The laboratory strain does not cause a similar effect. To activate neutrophils, bacterial opsonization is necessary. Differences in neutrophil NADH oxidase activation and ζ-potential for ZvL2-GLU and ZvL2-PA are associated with changes in membrane protein abundance, as demonstrated by differential 2D electrophoresis and LC-MS. The increase in ROS and cytokine production during the interaction of ZvL2-PA with neutrophils is associated with a rearrangement of the abundance of membrane proteins, which leads to the activation of Rcs and PhoP/Q signaling pathways and changes in the composition and/or modification of LPS. Certain isoforms of OmpA may play a role in the formation of the virulent phenotype of ZvL2-PA and participate in the activation of NADPH oxidase in neutrophils. [ABSTRACT FROM AUTHOR]

Background: Accelerated ageing is a method to assess seed storage performance by exposing the seeds to higher temperature and relative humidity to make seeds lose its vigour and viability quickly. The seed physiological parameters are then compared with the natural ageing to derive certain conclusions. But, the molecular pattern of seed deterioration under accelerated ageing varies compared to that of natural ageing. Methods: Chickpea, seed lots of variety JG-11 and Annigeri-1 were subjected to natural ageing in cloth bag with nine per cent initial seed moisture under ambient conditions. Simultaneously, representative sample of same seed lot were also aged at 41.C and 95±2% humidity up to 120 hours. Result: Both the ageing methods had significant negative effect on seed physiological and biochemical quality parameters. The germination potential of seeds tested after 48 hours of accelerated ageing (83.5%) was equal 10 months of natural ageing (85%) in JG-11, while also in Annigeri-1, 48 hours of accelerated ageing (78%) was nearly equal to 10 months of natural ageing (80%). The protein expression analyzed through 2D-PAGE at similar germination potential brought out by two independent ageing methods showed higher protein down regulation ratio (3.4) in accelerated ageing than in natural ageing (3.2) unveiling its rapidity in the seed deterioration process. Therefore, accelerated ageing can be used for predicting chickpea seed longevity. [ABSTRACT FROM AUTHOR]

Adhesive-invasive E. coli has been suggested to be associated with the development of Crohn’s disease (CD). It is assumed that they can provoke the onset of the inflammatory process as a result of the invasion of intestinal epithelial cells and then, due to survival inside macrophages and dendritic cells, stimulate chronic inflammation. In previous reports, we have shown that passage of the CD isolate ZvL2 on minimal medium M9 supplemented with sodium propionate (PA) as a carbon source stimulates and inhibits the adherent-invasive properties and the ability to survive in macrophages. This effect was reversible and not observed for the laboratory strain K12 MG1655. We were able to compare the isogenic strain AIEC in two phenotypes—virulent (ZvL2-PA) and non-virulent (ZvL2-GLU). Unlike ZvL2-GLU, ZvL2-PA activates the production of ROS and cytokines when interacting with neutrophils. The laboratory strain does not cause a similar effect. To activate neutrophils, bacterial opsonization is necessary. Differences in neutrophil NADH oxidase activation and ζ-potential for ZvL2-GLU and ZvL2-PA are associated with changes in membrane protein abundance, as demonstrated by differential 2D electrophoresis and LC-MS. The increase in ROS and cytokine production during the interaction of ZvL2-PA with neutrophils is associated with a rearrangement of the abundance of membrane proteins, which leads to the activation of Rcs and PhoP/Q signaling pathways and changes in the composition and/or modification of LPS. Certain isoforms of OmpA may play a role in the formation of the virulent phenotype of ZvL2-PA and participate in the activation of NADPH oxidase in neutrophils.

Lipoprotein lipase (LPL) is responsible for the intravascular catabolism of triglyceriderich lipoproteins and plays a central role in whole-body energy balance and lipid homeostasis. As such, LPL is subject to tissue-specific regulation in different physiological conditions, but the mechanisms of this regulation remain incompletely characterized. Previous work revealed that LPL comprises a set of proteoforms with different isoelectric points, but their regulation and functional significance have not been studied thus far. Here we studied the distribution of LPL proteoforms in different rat tissues and their regulation under physiological conditions. First, analysis by two-dimensional electrophoresis and Western blot showed different patterns of LPL proteoforms (i.e., different pI or relative abundance of LPL proteoforms) in different rat tissues under basal conditions, which could be related to the tissue-specific regulation of the enzyme. Next, the comparison of LPL proteoforms fromheart and brown adipose tissue between adults and 15-day-old rat pups, two conditions with minimal regulation of LPL in these tissues, yielded virtually the same tissue-specific patterns of LPL proteoforms. In contrast, the pronounced downregulation of LPL activity observed in white adipose tissue during fasting is accompanied by a prominent reconfiguration of the LPL proteoform pattern. Furthermore, refeeding reverts this downregulation of LPL activity and restores the pattern of LPL proteoforms in this tissue. Importantly, this reversible proteoform-specific regulation during fasting and refeeding indicates that LPL proteoforms are functionally diverse. Further investigation of potential differences in the functional properties of LPL proteoforms showed that all proteoforms exhibit lipolytic activity and have similar heparin-binding affinity, although other functional aspects remain to be investigated. Overall, this study demonstrates the ubiquity, differential distribution and specific regulation of LPL proteoforms in rat tissues and underscores the need to consider the existence of LPL proteoforms for a complete understanding of LPL regulation under physiological conditions. [ABSTRACT FROM AUTHOR]

Although one can trace the roots of proteomics well back into the 20th century, it is basically a discipline of the new millennium. At its outset, it was defined primarily by the technology available to analyze complex mixtures (basically 2D-gel electrophoresis, hybridizations/arrays and mass spectrometry) and what mainly set it aside from protein chemistry, that had flourished since the end of the second world war, was this use of unfractionated starting material as opposed to homogenous samples. Early on, two major new insights were quickly revealed: that the protein complement of cells was overwhelmingly involved in multiple protein–protein interactions and that it was nearly universally involved in a myriad of post-translational modifications. The revelations of the complex networks that result from these two phenomena have created a new understanding of cell biology that has affected our appreciation of such processes as transcription and translation, transmembrane signaling, differentiation, homeostasis and cell death. The development of these methods and strategies that principally characterize the field of proteomics depended heavily on the evolution of those that advanced protein chemistry, particularly during the last half of the twentieth century leading up to the elucidation of the human genome and will be briefly summarized in this article. In the last five decades of the 20th century, the original tools of protein chemistry combined and evolved to form the powerful largely mass spectrometry-based technology known as proteomics. This, together with the burgeoning science of genomics, has allowed the sensitive identification of proteins in complex samples, the principal targets of proteomics, including protein–protein interactions and post-translational modifications and their intricate roles in functional health and disease. [ABSTRACT FROM AUTHOR]

Objectives The present study aimed to evaluate the serum proteome of women with obsessive-compulsive disorder (OCD)/bipolar disorder (BP) compared to pure OCD subjects and healthy controls. Materials & Methods Serum proteome of women with OCD/BP, pure OCD individuals, and healthy controls were subjected to 2DE-based proteomics accompanied with MALDI-TOF-TOF mass spectrometry. Further evaluation of the identified protein spots with the significance of p<0.05 and fold≥1.5 was done by applying protein interaction mapping via Cytoscape v. 5.3.1 and its plugins. Results The results indicate that vitamin D binding protein (GC) and haptoglobin spots (HP) significantly changed expression in OCD and OCD/BP with different expression patterns. These identified spots may contribute to OCD/BP and act as differentially recognized biomarkers comparing pure OCD and OCD/BP. Conclusion The Findings imply that these proteins in the serum of the patients could be potential distinguishable biomarkers in clinical usage after related validation experiments. Therefore, this study provides a preliminary evaluation to understand OCD/BP proteome behavior better. [ABSTRACT FROM AUTHOR]

Oral carcinoma is one of the most vicious forms of cancer with a very low survival rate, as its patients often respond poorly to conventional chemotherapy. Presently several researchers are attempting to pursue an alternative to this therapy using natural products. Considering the promising strategy and induction of apoptosis to target the cancer cells, we evaluated the influence of a seaweed Padina gymnospora (15 µg/ml and 20 µg/ml) in enhancing apoptosis of oral cancer cells (KB-CHR-8–5) after 24-h incubation. The morphological changes indicating apoptosis were primarily assessed using a light microscope after which the apoptosis was confirmed by performing AO/EB staining method. Subsequently, MMP and ROS levels in the cells were assessed using Rh 123 and DCFH-DA staining procedures, respectively. All the above tests confirmed the ability of P. gymnospora to accelerate apoptosis in the oral cancer cells. As a next step, wide proteome analysis was performed where the proteins from P. gymnospora–treated cells were separated using the 2D electrophoresis technique and compared with that of control cells to isolate the differentially expressed proteins. This procedure resulted in the isolation of 10 proteins which were identified using MALDI-TOF/TOF MS, which established that most of the isolated proteins were part of the apoptotic process of the cell. The proteins identified are part of huge and complex pathways where it gets linked with many more genes which are also associated with apoptosis. Bioinformatics of these identified proteins was analyzed using STRING and PANTHER databases. These proteins contribute to cell apoptosis by affecting various functions, biological processes, and the synthesis of cellular components. PANTHER also demonstrated that these proteins belong to the classes of proteins that take part in several vital pathways of the cell among which the apoptotic pathway is the predominant one. [ABSTRACT FROM AUTHOR]

Simple Summary: Inulin-type fructans (ITFs), one of the best studied and most well-established prebiotic carbohydrates, have been shown to exert many health benefits to the host, including improvement in cardiovascular function. Various mechanisms are known to be involved in these processes, such as reduction of blood lipid levels by altering hepatic de novo lipogenesis, enhanced lipoprotein lipase activity in muscle, increased intestinal short-chain fatty acid production, elevated fecal excretion of bile salts and cholesterol and increased production of nitric oxide, known as endothelium-derived relaxing factor. Therefore, our overall aim was to assess the impact of complementary feeding with 4% of dried chicory root or 2% of native inulin on the aortic protein and gene changes in healthy nursery pigs. Our data clearly indicate that dietary inulin-type fructans have the potential to influence several structural and physiological alterations that are reflected both in the mRNA and protein levels in porcine aorta. In this study, two different ITFs sources were incorporated into a cereal-based diet to evaluate possible aortic protein and gene changes in nursery pigs. The animals were fed two different experimental diets from the 10th day of life, supplemented with either 4% of dried chicory root (CR) or with 2% of native inulin (IN). After a 40-day dietary intervention trial, pigs were sacrificed at day 50 and the aortas were harvested. Our data indicate that dietary ITFs have the potential to influence several structural and physiological changes that are reflected both in the mRNA and protein levels in porcine aorta. In contrast to our hypothesis, we could not show any beneficial effects of a CR diet on vascular functions. The direction of changes of several proteins and genes may indicate disrupted ECM turnover (COL6A1 and COL6A2, MMP2, TIMP3, EFEMP1), increased inflammation and lipid accumulation (FFAR2), as well as decreased activity of endothelial nitric oxide synthase (TXNDC5, ORM1). On the other hand, the IN diet may counteract a highly pro-oxidant environment through the endothelin–NO axis (CALR, TCP1, HSP8, PDIA3, RCN2), fibrinolytic activity (ANXA2), anti-atherogenic (CAVIN-1) and anti-calcification (LMNA) properties, thus contributing to the maintenance of vascular homeostasis. [ABSTRACT FROM AUTHOR]

Proteolysis is a post-translational modification (PTM) that affects the whole proteome. First regarded as only destructive, it is more precise than expected. It is finely regulated by other PTMs like phosphorylation. Aminopeptidase B (Ap-B), a M1 metallopeptidase, hydrolyses the peptide bond on the carbonyl side of basic residues at the NH 2 -terminus of peptides. 2D electrophoresis (2DE) was used to show that Ap-B is modified by phosphorylation. Detection of Ap-B by western blot after 2DE reveals several isoforms with different isoelectric points. Using alkaline phosphatase, Pro-Q Diamond phosphorylation-specific dye and kinase-specific inhibitors, we confirmed that Ap-B is phosphorylated. Phosphorylation can alter the structure of proteins leading to changes in their activity, localization, stability and association with other interacting molecules. We showed that Ap-B phosphorylation might delay its turnover. Our study illustrates the central role of the crosstalk between kinases and proteases in the regulation of many biological processes. • The metallopeptidase, hydrolyses the peptide bond on the carbonyl side of basic residues at the NH 2 -terminus of peptides. • Phosphorylation is responsible for the regulation of many proteases. • After 2D electrophoresis, Ap-B presents a "train of spots" signature which is specific of phosphorylation. • Specific dyes and kinase inhibitors were used to show that Ap-B is phosphorylated on serine, threonine and tyrosine residues. • Ap-B phosphorylation seems to delay its turnover. [ABSTRACT FROM AUTHOR]

Deep ocean water (DOW) is rich in mineral nutrients, and our previous studies have confirmed that it promotes fungal growth and metabolite production. Cordycepin is a functional metabolite of Cordyceps with anti-inflammatory and antitumor properties. Although Cordyceps militaris can produce a large amount of cordycepin when DOW is used as the culture water, no study has explored the relevant regulatory mechanism through which DOW increases the adenosine and cordycepin contents. Therefore, in this study, we used 2D electrophoresis and proteomic analysis to investigate the effects of DOW with different mineral compositions on proteins in the fruiting bodies of C. militaris and to explore how DOW regulates cordycepin biosynthesis. The results indicated that the use of DOW with high magnesium and calcium contents can increase the production of cordycepin from 6.62 mg/g to 11.40 mg/g by C. militaris fruiting bodies. According to the 2D electrophoresis and proteomic analysis results of the fruiting body proteins of C. militaris cultured with DOW with different concentrations, the expression of 17 and 9 proteins was proportional to the cordycepin content and the concentration of DOW, respectively. Magnesium, sulfate, and other minerals of DOW stimulated the production of cordycepin by promoting the glycolytic pathway, producing more ATP and adenosine. It also increased the production of nucleic acids and adenosine by promoting the pentose phosphate pathway and methionine cycle, thereby promoting the growth and cordycepin production of fruiting bodies. [ABSTRACT FROM AUTHOR]

Aim/Background: Cancer is still known to be a devastating disease condition. Even after getting advancements in multimodality therapies, the success rate with respect to patient survival is very limited. Hence there is an emerging need to find alternative medication strategies for cancer treatment. Recently seaweeds are getting their significance due to its tremendous medicinal properties. Among different category of seaweeds, Padina gymnospora which is a brown seaweed is known to possess many important bioactivities including antimicrobial activity, anti-oxidative property and anti-inflammatory. Role of this seaweed in treating oral cancer cells is not much explored in detail. Hence, this study aims to understand the significance of Padina gymnospora in oral cancer cell line -- SAS. Materials and Methods: The ethanol extract of Padina was treated to oral cancer cell line -- SAS and incubated for 24 hr. After incubation, MTT assay was performed to find the efficiency of the extract. Further the cells were taken for protein extraction and subjected to SDS PAGE, two-dimensional electrophoresis (2D) coupled with tandem mass spectrometry approach. The differentially expressed proteins were further identified and subjected to bioinformatic analysis. Results and Conclusion: It was observed that pro apoptotic proteins including BIK, BAK1, BID, BAD and CASP8 were highly upregulated in the Padina treated samples compared with untreated control samples. This data clearly demonstrates that Padina gymnospora induces the apoptosis in the oral cancer cells. This study paves us the way to explore more about the therapeutic potential of Padina gymnospora in treating the oral cancer cells. [ABSTRACT FROM AUTHOR]

The illicit use of dexamethasone and other glucocorticoids for cattle fattening in livestock production has been widely described; evidence for illegal treatments can be obtained by direct or indirect detection. In our previous study, we applied two-dimensional electrophoresis (2DE) to identify plasma protein markers of dexamethasone administration in veal calves. Comparison of 2DE maps obtained from blood samples before and after treatment showed the disappearance of two protein spots identified as serum paraoxonase/arylesterase 1 precursor (PON1). In the present study, we validated PON1 as a marker by analysing a larger number of samples treated with dexamethasone for illicit use. Analysis of samples from experimental treatment with other glucocorticoids, androgens and oestrogens confirmed that their influence on PON1 could be excluded. The specificity of the PON1 protein marker was verified on expected negative field samples to exclude interfering factors. However, there is poor statistical evidence to support a significant association between the outcome of PON1 and the considered variables. The results on field samples were compared with histological examination of the thymus as a biomarker of corticosteroid treatment monitored in the Italian histological plan for the control of growth promoters in animals. Two suspect cases were identified from two Piedmont farms where other animals had tested positive at histological examination. In conclusion, the absence of PON1 in the plasma of veal calves can indirectly reveal illicit dexamethasone treatment in individual animals and so identify suspect farms for further investigation. It is effective in a period ranging from 3 to about 10 days from illicit treatment, covering a time span that goes beyond the limits of official chemical controls and preceding histological controls on the thymus of slaughtered animals. PON1 detection in plasma can be coupled with other tests to identify illegal dexamethasone use on veal calf farms. [ABSTRACT FROM AUTHOR]

Background : Bee venom has been used as a therapeutic compound for various human diseases in oriental medicine; however, it can induce anaphylaxis in hypersensitive patients during treatment. Anaphylaxis is an acute allergic reaction that occurs after allergen exposure. IgE is released from immune-related cells such as mast cells and basophils during anaphylaxis. Various inflammatory mediators are also released into the bloodstream during the acute response. Objectives: We aimed to identify specific proteins from bee venom-hypersensitive patients. Methods : We analyzed the blood serum of control and bee venom-hypersensitive patients using two-dimensional (2D) electrophoresis. Results : An interesting protein spot with a molecular size of 10 kDa was identified at an isoelectric point (p.I.) of 5.5. Spots detected both before and after sweet bee venom therapy were not proteins induced by sweet bee venom. The 10 kDa protein was identified as the cleaved form of haptoglobin through liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Statistical analysis indicated that the presence of the spot was highly significant in the bee venom-hypersensitive group. Conclusion : The findings suggest that cleaved haptoglobin may be a significant diagnostic protein for anaphylaxis. In addition, a high incidence of bee venom hypersensitivity may be associated with the haptoglobin genotype.

Copyright of Revista Cientifica de la Facultade de Veterinaria is the property of Universidad del Zulia, Facultad de Ciencias Veterinarias and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Description of the subject. Commercialization requires the storage of meat for a period of time according to the distribution and supply chain, usually under refrigeration and vacuum packaging. During this stage, many biochemical reactions can modify the sensory and physicochemical properties of meat. Proteolysis is one of the most important of these reactions as proteins are the building blocks of muscle, and their degradation affects tenderness. Objectives. The objectives of this study were to use multiple factorial analyses to correlate the physicochemical, sensory, and proteolytic changes in ovine meat during refrigeration. Method. Each loin was separated and randomly assigned to day 3, 5, or 14. Sensory panel evaluated the meat for appearance, odor and texture parameters. Warner-Bratzler shear force and texture profile analysis were evaluated. Similarly, protease activity was evaluated. Samples for 2D-electrophoresis and western blotting were collected at days 3, 5, and 14 post mortem. The results were analyzed by multiple factorial analyses. Results. The relative protein levels of desmin, vinculin, and myosin were found to decrease with time. Multiple factor analysis showed that the Warner-Bratzler shear force (WBSF) was highly correlated with the relative desmin concentration, the relative vinculin concentration, and acidic protease activity. The activities of acidic and neutral proteases were positively correlated with myosin level on day 3, but were negatively correlated with manual tenderness and myosin level on days 5 and 14. Conclusions. Therefore, desmin, myosin and vinculin, which correlate with sensory and physicochemical parameters during refrigerated storage, may prove useful as markers for tenderness.

The aim of this study was to gain insight into pancreatic proteome changes induced by dexamethasone treatment. Sixweek-old male ICR mice were subcutaneously injected with or without dexamethasone (10 mg/kg body weight; n= 5-6 mice per group) for 4 weeks. Body weight, food intake, and fasting blood glucose were measured at baseline and at the end of 4 weeks. The mice were sacrificed and pancreases were removed, frozen, and stored at -75˚C until analysis. Proteins were extracted, twodimensional gel electrophoresis was performed and proteome profiles were examined with colloidal coomassie brilliant blue staining. Statistically significant protein spots (ANOVA, p

In this study, two different ITFs sources were incorporated into a cereal-based diet to evaluate possible aortic protein and gene changes in nursery pigs. The animals were fed two different experimental diets from the 10th day of life, supplemented with either 4% of dried chicory root (CR) or with 2% of native inulin (IN). After a 40-day dietary intervention trial, pigs were sacrificed at day 50 and the aortas were harvested. Our data indicate that dietary ITFs have the potential to influence several structural and physiological changes that are reflected both in the mRNA and protein levels in porcine aorta. In contrast to our hypothesis, we could not show any beneficial effects of a CR diet on vascular functions. The direction of changes of several proteins and genes may indicate disrupted ECM turnover (COL6A1 and COL6A2, MMP2, TIMP3, EFEMP1), increased inflammation and lipid accumulation (FFAR2), as well as decreased activity of endothelial nitric oxide synthase (TXNDC5, ORM1). On the other hand, the IN diet may counteract a highly pro-oxidant environment through the endothelin–NO axis (CALR, TCP1, HSP8, PDIA3, RCN2), fibrinolytic activity (ANXA2), anti-atherogenic (CAVIN-1) and anti-calcification (LMNA) properties, thus contributing to the maintenance of vascular homeostasis.

Deep ocean water (DOW) is rich in mineral nutrients, and our previous studies have confirmed that it promotes fungal growth and metabolite production. Cordycepin is a functional metabolite of Cordyceps with anti-inflammatory and antitumor properties. Although Cordyceps militaris can produce a large amount of cordycepin when DOW is used as the culture water, no study has explored the relevant regulatory mechanism through which DOW increases the adenosine and cordycepin contents. Therefore, in this study, we used 2D electrophoresis and proteomic analysis to investigate the effects of DOW with different mineral compositions on proteins in the fruiting bodies of C. militaris and to explore how DOW regulates cordycepin biosynthesis. The results indicated that the use of DOW with high magnesium and calcium contents can increase the production of cordycepin from 6.62 mg/g to 11.40 mg/g by C. militaris fruiting bodies. According to the 2D electrophoresis and proteomic analysis results of the fruiting body proteins of C. militaris cultured with DOW with different concentrations, the expression of 17 and 9 proteins was proportional to the cordycepin content and the concentration of DOW, respectively. Magnesium, sulfate, and other minerals of DOW stimulated the production of cordycepin by promoting the glycolytic pathway, producing more ATP and adenosine. It also increased the production of nucleic acids and adenosine by promoting the pentose phosphate pathway and methionine cycle, thereby promoting the growth and cordycepin production of fruiting bodies.

Radiated waves from mobile phones are in the Radio Frequency (RF) range, so those are unable to cause ionization and electron excitation. RF that are produced and emitted from sender antenna on the surface of mobile phone systems, ranges from 30 up to 300 GHz in electromagnetic wave spectrum. There is no complete information about the effects of RF on protein changes. For this purpose, we have examined the effects of RF waves on the changes in protein expression pattern of the brain. In this study, 10 heads of male Wistar rats, weighing between 200-250 g, were exposed to transmission antenna in 100-180 meter distance. Then, proteome of RF900 GHz exposed group and control group were extracted. Two-dimensional gel electrophoresis evaluation and proteomics analysis was performed. Results showed altered proteome pattern due to radiation. Tissue exposed to nonstandard waves showed similar pattern of changes in the expression of some proteins, which have key roles in the induction of apoptosis. The expressions of key proteins including Bcl-2-A1, Bid, Neurofilament and Cytochrome Oxidase were decreased. Expression or suppression of apoptosis related proteins such as BCL-2 in rat brain proteome exposed to standard RF900 MHz (at 180m and beyond), can serve as a biomarker of brain activity, memory and sleep. RF radiated from transmission antenna in urban and standard spaces may not be carcinogenic, but, individuals exposed in nonstandard distances to those antennas (less than 180meter) may be at risk. HIGHLIGHTS •Radio Frequency (RF) waves can affect the protein expression profile of the cell. •Exposure to nonstandard RF waves changed the expression profile of apoptosis key proteins. •Changes in the expression of Bcl-2-A1, Bid, Neurofilament and Cytochrome Oxidase under the RF was evident.

Seedlings of spring wheat (Triticum aestivum L.) cultivars, Ethos and Zebra, differing in drought tolerance were dehydrated to reach a water saturation deficit (WSD) in leaves ~15, 30, and 50 %. Ethos, the drought tolerant cultivar, dried slower in comparison with Zebra and regrew in 70 % upon rehydration. The effect of dehydration on photosystem II was evaluated by Chl a fluorescence (OJIP transients). The inflection point of double normalized curves (ΔWOJ) calculated for Ethos was negative for seedlings with 15 % WSD, nearly zero for those with 30 % WSD, and about +0.05 for those with 50 % WSD. In case of Zebra, the 15 % WSD already induced a positive ΔWOJ (+0.05) and 50 % WSD maximized it to +0.10, which is a sign of drought susceptibility. The proteomic studies revealed, that among identified 850 spots, 80 protein spots were differentially expressed during dehydration. The differentially expressed proteins of the drought tolerant cultivar indicated the protection of the photosynthetic apparatus and proteome rebuilding in response to drought. In the drought susceptible cultivar, protection of proteins and membranes and partial scavenging reactive oxygen species appeared.

Background/Aim: The E6 genotypic variants of HPV 16 identified in lesions of women with cervical cancer (CC) in Southern Mexico include the E-G350, AAa, AAc, EC188/G350, and E-A176/G350. Transcriptomic analysis cells transfected with those variants showed to induce differential expression of the host genes involved in the development of CC. The aim of this work was to understand how the over-expression of the E6 oncoprotein and its variants can induce molecular mechanisms that lead to more aggressive HPV 16 phenotypes in cervical cancer and which proteins could be associated with the process. Materials and Methods: Total extracts from C33A, C33A mock, C33A AAa, C33A E-C188/G350, C33A E-A176/G350, and C33A Eprototype cells were analyzed using 2D electrophoresis, PDQuest software and mass spectrometry. Validation of results was performed through qPCR. Results: Statistically significant differential expression of 122 spots was detected, 12 of the identified proteins were associated with metabolism and metabolic programming. Out of these CCT8, ENO and ALDH1A were further validated. Conclusion: CCT8 and ALDH1A were found to be over-expressed in C33A AAa and C33A E-A176/G350, compared to the E prototype. Both proteins could be associated with a most aggressive phenotype due to their relationship with metabolism, protein folding and stemness, mechanisms associated to E6 that could be useful in the design of new therapies. [ABSTRACT FROM AUTHOR]

Crohn’s disease (CD) is a severe chronic immune-mediated granulomatous inflammatory disease of the gastrointestinal tract. The mechanisms of CD pathogenesis remain obscure. Metagenomic analysis of samples from CD patients revealed that several of them have the elevated level of Escherichia coli with adhesive–invasive phenotype (AIEC). Previously, we isolated an E. coli strain CD isolate ZvL2 from a patient with CD, which features AIEC phenotype. Here, we demonstrate that prolonged growth on propionate containing medium stimulates virulent properties of CD isolate ZvL2, while prolonged growth on glucose reduces these properties to levels indistinguishable from laboratory strain K-12 MG1655. Propionate presence also boosts the ability of CD isolate ZvL2 to penetrate and colonize macrophages. The effect of propionate is reversible, re-passaging of CD isolate on M9 medium supplemented with glucose leads to the loss of its virulent properties. Proteome analysis of CD isolate ZvL2 growth in medium supplemented with propionate or glucose revealed that propionate induces expression porins OmpA and OmpW, transcription factors PhoP and OmpR, and universal stress protein UspE, which were previously found to be important for macrophage colonization by enteropathogenic bacteria.

For adaptation to stressful conditions, Mycobacterium tuberculosis (Mtb) is prone to transit to a dormant, non-replicative state, which is believed to be the basis of the latent form of tuberculosis infection. Dormant bacteria persist in the host for a long period without multiplication, cannot be detected from biological samples by microbiological methods, however, their “non-culturable” state is reversible. Mechanisms supporting very long capacity of mycobacteria for resuscitation and further multiplication after prolonged survival in a dormant phase remain unclear. Using methods of 2D electrophoresis and MALDI-TOF analysis, in this study we characterized changes in the proteomic profile of Mtb stored for more than a year as dormant, non-replicating cells with a negligible metabolic activity, full resistance to antibiotics, and altered morphology (ovoid forms). Despite some protein degradation, the proteome of 1-year-old dormant mycobacteria retained numerous intact proteins. Their protein profile differed profoundly from that of metabolically active cells, but was similar to the proteome of the 4-month-old dormant bacteria. Such protein stability is likely to be due to the presence of a significant number of enzymes involved in the protection from oxidative stress (katG/Rv1908, sodA/Rv3846, sodC/Rv0432, bpoC/Rv0554), as well as chaperones (dnaJ1/Rv0352, htpG/Rv2299, groEL2/Rv0440, dnaK/Rv0350, groES/Rv3418, groEL1/Rv3417, HtpG/Rv2299c, hspX/Rv2031), and DNA-stabilizing proteins. In addition, dormant cells proteome contains enzymes involved in specific metabolic pathways (glycolytic reactions, shortened TCA cycle, degradative processes) potentially providing a low-level metabolism, or these proteins could be “frozen” for usage in the reactivation process before biosynthetic processes start. The observed stability of proteins in a dormant state could be a basis for the long-term preservation of Mtb cell vitality and hence for latent tuberculosis.

Channel catfish (Ictalurus punctatus) is the top species produced in US aquaculture and motile Aeromonas septicemia, caused by virulent Aeromonas hydrophila (vAh), is one of the most severe diseases that afflict catfish farms. Previously, vaccination of fish with extracellular proteins (ECP) of vAh was shown to produce a robust antibody-mediated immune response against vAh infection. In this study, we analyzed IgM transcripts that were differentially expressed in the head kidney and liver of ECP-immunized and mock-immunized (control) fish with emphasis on a variable domain of heavy chain. Quantitative PCR analysis indicated that immunized fish produced significantly more IgM transcripts than control fish. Full-length IgM heavy chain cDNA was cloned, which encoded typical IgM peptide, including signal peptide, variable domain (VH), constant domain (CH), and carboxyl terminal peptide. Great sequence diversity was revealed in a VH segment, with the third complementarity diversity region (CDR3) being most variable. Using germline VH gene grouping method, variants (clones) of VH characterized in this study belonged to nine VH families. The most unique variants (approximately 49%) were found in the VH2 family. Vaccinated fish apparently had more unique variants than in the control fish. There were 62% and 79% of unique variants in the head kidney and liver of vaccinated fish, respectively, while 44% and 27% unique variants in the head kidney and liver of control fish, respectively. Among the unique variants in VH2 family, approximately 87% of them were found in vaccinated fish. Two-dimensional gel electrophoresis of semi-purified IgM protein confirmed that matured IgM protein was as variable as IgM transcripts identified in this study, with isoelectric points crossing from 6 to 10. Results of this study provided insight into the molecular and genetic basis of antibody diversity and enriched our knowledge of the complex interplay between antigens and antibodies in Ictalurid catfish.

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The aim of this study was to gain insight into pancreatic proteome changes induced by dexamethasone treatment. Sixweek-old male ICR mice were subcutaneously injected with or without dexamethasone (10 mg/kg body weight; n= 5-6 mice per group) for 4 weeks. Body weight, food intake, and fasting blood glucose were measured at baseline and at the end of 4 weeks. The mice were sacrificed and pancreases were removed, frozen, and stored at-75˚C until analysis. Proteins were extracted, twodimensional gel electrophoresis was performed and proteome profiles were examined with colloidal coomassie brilliant blue staining. Statistically significant protein spots (ANOVA, p<0.05) with 1.2-fold difference were excised and analyzed by LCMS/MS. Proteomic data were evaluated using Western blot analysis. High-dose dexamethasone induced hyperglycemia, reduced body weight, reduced food intake, and altered pancreatic proteins in mice. The affected proteins mainly belonged to cell redox homeostasis, metabolism, endoplasmic reticulum stress, translation, and signaling pathways. [ABSTRACT FROM AUTHOR]

For adaptation to stressful conditions, Mycobacterium tuberculosis (Mtb) is prone to transit to a dormant, non-replicative state, which is believed to be the basis of the latent form of tuberculosis infection. Dormant bacteria persist in the host for a long period without multiplication, cannot be detected from biological samples by microbiological methods, however, their "non-culturable" state is reversible. Mechanisms supporting very long capacity of mycobacteria for resuscitation and further multiplication after prolonged survival in a dormant phase remain unclear. Using methods of 2D electrophoresis and MALDI-TOF analysis, in this study we characterized changes in the proteomic profile of Mtb stored for more than a year as dormant, non-replicating cells with a negligible metabolic activity, full resistance to antibiotics, and altered morphology (ovoid forms). Despite some protein degradation, the proteome of 1-year-old dormant mycobacteria retained numerous intact proteins. Their protein profile differed profoundly from that of metabolically active cells, but was similar to the proteome of the 4-month-old dormant bacteria. Such protein stability is likely to be due to the presence of a significant number of enzymes involved in the protection from oxidative stress (katG/Rv1908, sodA/Rv3846, sodC/Rv0432, bpoC/Rv0554), as well as chaperones (dnaJ1/Rv0352, htpG/Rv2299, groEL2/Rv0440, dnaK/Rv0350, groES/Rv3418, groEL1/Rv3417, HtpG/Rv2299c, hspX/Rv2031), and DNA-stabilizing proteins. In addition, dormant cells proteome contains enzymes involved in specific metabolic pathways (glycolytic reactions, shortened TCA cycle, degradative processes) potentially providing a low-level metabolism, or these proteins could be "frozen" for usage in the reactivation process before biosynthetic processes start. The observed stability of proteins in a dormant state could be a basis for the long-term preservation of Mtb cell vitality and hence for latent tuberculosis. [ABSTRACT FROM AUTHOR]

Drought is a major abiotic stress that negatively influences crop yield. Breeding strategies for improved drought resistance require an improved knowledge of plant drought responses. We therefore applied drought to barley recombinant inbred lines and their parental genotypes shortly before tillering. A large-scale proteomic analysis of leaf and root tissue revealed proteins that respond to drought in a genotype-specific manner. Of these, Rubisco activase in chloroplast, luminal binding protein in endoplasmic reticulum, phosphoglycerate mutase, glutathione S-transferase, heat shock proteins and enzymes involved in phenylpropanoid biosynthesis showed strong genotype×environment interactions. These data were subjected to genetic linkage analysis and the identification of proteomic QTLs that have potential value in marker-assisted breeding programs. [ABSTRACT FROM AUTHOR]

Abstract Atherosclerosis and cancer are the leading causes of mortality around the world that share common pathogenic pathways. The aim of this study is the investigation of the protein profile of atherosclerotic plaque in order to find similar biomarker between cancer and atherosclerosis. The small pieces of human coronary artery containing advanced atherosclerotic plaque is obtained from patients during bypass surgery. Structural characterization of type V plaque, including fibrous connective tissue, necrotic lipid core, cholesterol clefts and calcium deposits are performed using high resolution transmission electron microscopy (HR-TEM). The protein profile of atherosclerosis plaque is also analyzed using 2-dimensional electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF). TEM analysis shows that vascular smooth muscle cells (VSMCs) exhibit different and uncommon morphologies in atherosclerotic plaque which is correlated to the proliferative state of the cells. The proteomics analysis reveals proteins related to atherosclerosis formation including Mimecan, Ras Suppressor Protein-1 (RSUP-1) and Cathepsin D which identified as biomarker of cancerous tumors. The expression of Mimecan and RSUP-1 is down-regulated in atherosclerotic plaque while the expression of Cathepsin D is up-regulated. These data support that atherosclerotic plaque presents some degree of tumorgenesis with the significant activity of VSMCs as the key player in atherogenesis. Highlights • The protein profile of human atherosclerosis plaque is a analyzed using 2D electrophoresis and MALDI-TOF. • The proteomics analysis reveals proteins related to atherosclerosis formation including Mimecan, RSUP-1 and Cathepsin D. • The expression of Mimecan and RSUP-1 is down-regulated in atherosclerotic plaque. • The expression of Cathepsin D is up-regulated in atherosclerotic plaque. • The data support similar biomarker between cancer and atherosclerosis. [ABSTRACT FROM AUTHOR]

Purpose: Uveitis is a cause for concern in the developing countries like India. Its poor diagnosis and lack of proper therapeutics often cause blindness in children and young adults. Moreover, the exact mechanism of pathogenesis of different types of uveitis is still elusive. Modern proteomic techniques are found to be advantageous for an in‐depth understanding of the ocular physiology using proteomic diversity. Our aim was to identify unique proteins involved in the pathogenesis of autoimmune or noninfectious uveitis. Methods: Vitreous fluid samples (n = 90) were obtained from infectious (N = 34) and noninfectious (N = 56) uveitis patients, and their protein profiles were compared by analysing sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE) and 2D electrophoresis. Unique proteins were identified through matrix‐assisted laser desorption ionization–time‐of‐flight mass spectrometry (MALDI‐TOF MS) and further studied for pathway analysis. Results: Protein spots having different molecular weights were observed in noninfectious vitreous fluid samples. Enzymatic digestion of these spots after MALDI‐TOF MS analysis revealed different proteins. We identified 25 different proteins through SDS‐PAGE and 22 through 2D electrophoresis. 50% of the proteins from SDS‐PAGE were associated with heterotrimeric G‐protein signalling pathway–rod outer segment phototransduction. 50% proteins from SDS‐PAGE and 20% from 2D electrophoresis revealed association with de novo purine biosynthesis. Carbonic anhydrase 1 and serpin B3 were found to be common in both analyses. Conclusion: High‐throughput proteomic and pathway analyses have exposed the potential association of these proteins with autoimmune pathogenesis in uveitis. The exact role of most of the proteins in autoimmune uveitis is yet to be unfurled. [ABSTRACT FROM AUTHOR]

Peptidase therapy is suggested to be effective to minimize gliadin toxicity in celiac disease (CD). Hence, present study deals with gliadin-hydrolysing peptidases. The efficient peptidase from the Bacillus tequilensis was purified using ammonium sulfate fractionation and preparative electrophoresis. Analysis of in-solution and in-gel hydrolysis of gliadin using one and two-dimensional SDS-PAGE revealed nearly complete hydrolysis of gliadin peptides after 180 min of incubation with B. tequilensis protease. Purified peptidase was found to be stable at acidic (pH 3.5) to neutral (pH 7.2) pH range. The molecular mass and isoelectric point of the peptidase were observed around 29 kDa and 5.2, respectively. The internal protein sequence obtained through mass spectrometric analysis suggested that peptidase might belong to peptidase S9 family known for prolyl-specific peptidases. This study recommends the possible applicability of this peptidase for elimination of immunotoxic gliadin peptides and may prove useful in CD treatment. [ABSTRACT FROM AUTHOR]

In dry savannahs of West-Africa, the malarial mosquitoes of the Anopheles gambiae sensu stricto complex annually survive the harsh desiccating conditions of the dry season. However, the physiological and biochemical mechanisms underlying how these mosquitoes survive such desiccating conditions are still undefined, and controversial. In this context, we provide the first work examining both proteomic and metabolomic changes in the two molecular forms of A. gambiae s.s (M and S forms) experimentally exposed to the rainy and dry season conditions as they experience in the field. Protein abundances of the mosquitoes were measured using a two-dimensional fluorescence difference gel electrophoresis (2D DIGE) coupled with a matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) and tandem mass spectrometry (MS) for protein identification. These assays were conducted by Applied Biomics (http://www.appliedbiomics.com, Applied Biomics, Inc. Hayward, CA, USA), and the mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository with the dataset identifier PXD000294. The metabolomic analysis was conducted using both Acquity UPLC® system (for amino acid identification), and a gas-chromatography-mass spectrometry platform (for sugars identification). Metabolomic fingerprintings were assessed in the University of Rennes 1, UMR CNRS 6553 EcoBio (France). A detailed interpretation of the obtained data can be found in Hidalgo et al. (2014) [1] (Journal of Insect Physiology (2014)).

Somatic embryogenesis (SE) is a cell differentiation process by which a somatic cell changes its genetic program and develops into an embryonic cell. Investigating this process with various explant sources in vitro has allowed us to trace somatic embryo development from germination to plantlets and has led to the generation of new technologies, including genetic transformation, endangered species conservation, and synthetic seed production. A transcriptome data comparison from different stages of the developing somatic embryo has revealed a complex network controlling the somatic cell’s fate, suggesting that an interconnected network acts at the protein level. Here, we discuss the current progress on SE using proteomic-based data, focusing on changing patterns of proteins during the establishment of the somatic embryo. Despite the advanced proteomic approaches available so far, deciphering how the somatic embryo is induced is still in its infancy. The new proteomics techniques that lead to the quantification of proteins with different abundances during the induction of SE are opening this area of study for the first time. These quantitative differences can elucidate the different pathways involved in SE induction. We envisage that the application of these proteomic technologies can be pivotal to identifying proteins critical to the process of SE, demonstrating the cellular localization, posttranslational modifications, and turnover protein events required to switch from a somatic cell to a somatic embryo cell and providing new insights into the molecular mechanisms underlying SE. This work will help to develop biotechnological strategies for mass production of quality crop material.

Mycobacteria are able to form dormant cells, which survive for a long time without multiplication. The molecular mechanisms behind prolonged survival of dormant cells are not fully described. In particular, little information is known on biochemical processes which might take place in cells under dormancy. To gain insight into this problem, Mycobacterium smegmatis cells in deep dormant state were obtained after gradual acidification of the growth medium in prolonged stationary phase followed by 1 month of storage at room temperature. Such cells were characterized by low metabolic activity, including respiration, resistance to antibiotics, and altered morphology. The protein composition of cytoplasm and membrane fractions obtained from active and dormant cells were compared by 2D electrophoresis. Almost half of the proteins found in the proteome of dormant cells were absent in that of active cells. This result differs significantly from published results obtained in other studies employing different models of mycobacterium dormancy. This discrepancy could be explained by a deeper dormancy developed in the present model. A feature of a “dormant proteome” is high representation of enzymes involved in glycolysis and defense systems that inactivate or detoxify reactive oxygen and nitrogen species, aldehydes, and oxidized lipids. Dormant mycobacteria are enriched by degradative enzymes, which could eliminate damaged molecules, or the products of such degradation could be reutilized by the cell during prolonged storage. We suggest that some enzymes in dormant cells are inactive, having been used upon transition to the dormant state, or proteins stored in dormant cells for further cell reactivation. At the same time, some proteins could be functional and play roles in maintenance of cell metabolism, albeit at a very slow rate. This study provides a clue as to which biochemical processes could be active under dormancy to ensure long-term viability of dormant mycobacteria.

Abstract We document the changes in sera and liver proteome during experimental hepatic fibrosis (HF) and its protection by resveratrol (Rsvtrl) in rats. HF was induced by N ′-Nitrosodimethylamine (NDMA) administration thrice a week for 21-days. Compared to the control group, significant (P < 0.05) elevations in ALT, AST, ALP, γGT and bilirubin occurred during fibrosis. H&E, IHC for α-SMA and reticulin stainings of liver demonstrated that collagen amassing and HF were concurrent. Rsvtrl-supplementation refurbished hepatic architecture and lessened collagen deposition. 2DE proteome analyses showed ~398 (pH, 3–10) and ~129 signatures (pH, 4–7) within 5–201 kDa range in liver and sera, respectively. Nearly, 45 spots in liver and 18 in sera were differentially expressed in fibrotic and Rsvtrl-supplemented animals. MALDI−TOF–MS/MS analysis revealed at least ten signatures as the potential biomarkers for HF, because of the significant changes (>2 fold) in their expression. Out of them, two signatures have not so far been characterized and we have accomplished it in this study. Our proteomics data provides new evidence on an overall involvement of carbohydrate, lipid, Ca2+ signaling and oxidative pathways during NDMA-induced HF. More importantly, the oxidative pathway appears to be the principal mediator in the NDMA-induced HF as well as the hepato-protection by Rsvtrl. Highlights • Proteomic study on fibrotic (HF) and resveratrol-supplemented rats is carried out. • 2D-MALDI-MS analysis revealed 10 major protein spots including 2 uncharacterized. • Carbohydrate, lipid and Ca2+ signaling pathway are inter-linked during HF. • Oxidative pathway appears principal to mediate liver damage. • Proteomic evidences suggest strong hepatoprotective potential of resveratrol. [ABSTRACT FROM AUTHOR]

Mycobacteria are able to form dormant cells, which survive for a long time without multiplication. The molecular mechanisms behind prolonged survival of dormant cells are not fully described. In particular, little information is known on biochemical processes which might take place in cells under dormancy. To gain insight into this problem, Mycobacterium smegmatis cells in deep dormant state were obtained after gradual acidification of the growth medium in prolonged stationary phase followed by 1 month of storage at room temperature. Such cells were characterized by low metabolic activity, including respiration, resistance to antibiotics, and altered morphology. The protein composition of cytoplasm and membrane fractions obtained from active and dormant cells were compared by 2D electrophoresis. Almost half of the proteins found in the proteome of dormant cells were absent in that of active cells. This result differs significantly from published results obtained in other studies employing different models of mycobacterium dormancy. This discrepancy could be explained by a deeper dormancy developed in the present model. A feature of a "dormant proteome" is high representation of enzymes involved in glycolysis and defense systems that inactivate or detoxify reactive oxygen and nitrogen species, aldehydes, and oxidized lipids. Dormant mycobacteria are enriched by degradative enzymes, which could eliminate damaged molecules, or the products of such degradation could be reutilized by the cell during prolonged storage. We suggest that some enzymes in dormant cells are inactive, having been used upon transition to the dormant state, or proteins stored in dormant cells for further cell reactivation. At the same time, some proteins could be functional and play roles in maintenance of cell metabolism, albeit at a very slow rate. This study provides a clue as to which biochemical processes could be active under dormancy to ensure long-term viability of dormant mycobacteria. [ABSTRACT FROM AUTHOR]

In the present study, 155 Nellore cattle were genotyped for the CAPN4751 and UOGCAST polymorphisms and phenotyped for shear force (SF) at 1, 7 and 14 days aging. The effects of different genotypic combinations were evaluated on the Longissimus muscle proteomic profile using 2DE and mass spectrometry. A significant association was found between genotypes for UOGCAST and CAPN4751 and meat tenderness. The CC genotype for both markers was favorable for lesser shear force than TT. A total of 40 spots showed significant differential expression profiles (P < 0.05), of which eight had a main effect for the CAPN4751 marker, 11 for UOGCAST, two for both markers, and 19 had interactions between markers, including myosin (MYL1, MYL2, MYLPF and MYL6B), actin (ACTA1 and CAPZβ), troponin (TNNT1 and TNNT3) and heat shock proteins (HSPB6, HSPB1 and HSP70-2). The results demonstrated that UOGCAST and CAPN4751 genotypes led to variability on the expression of proteins that are involved in muscle metabolism, and consequently affect meat tenderness. [ABSTRACT FROM AUTHOR]

Abstract: The protein content and allergen composition was studied of cashews from 8 different origins (Benin, Brazil, Ghana, India, Ivory Coast, Mozambique, Tanzania, Vietnam), subjected to different in‐shell heat treatments (steamed, fried, drum‐roasted). On 2D electrophoresis, 9 isoforms of Ana o 1, 29 isoforms of Ana o 2 (11 of the acidic subunit, 18 of the basic subunit), and 8 isoforms of the large subunit of Ana o 3 were tentatively identified. Based on 1D and 2D electrophoresis, no difference in allergen content (Ana o 1, 2, 3) was detected between the cashews of different origins (P > 0.5), some small but significant differences were detected in allergen solubility between differently heated cashews. No major differences in N‐ and C‐terminal microheterogeneity of Ana o 3 were detected between cashews of different origins. Between the different heat treatments, no difference was detected in glycation, pepsin digestibility, or IgE binding of the cashew proteins. [ABSTRACT FROM AUTHOR]

Mitochondria are exposed to reactive nitrogen species under physiological conditions and even more under several pathologic states. In order to reveal the mechanism of these processes we studied the effects of peroxynitrite on isolated beef heart mitochondria in vitro. Peroxynitrite has the potential to nitrate protein tyrosine moieties, break the peptide bond, and eventually release the membrane proteins into the solution. All these effects were found in our experiments. Mitochondrial proteins were resolved by 2D electrophoresis and the protein nitration was detected by immunochemical methods and by nano LC-MS/MS. Mass spectrometry confirmed nitration of ATP synthase subunit beta, pyruvate dehydrogenase E1 component subunit beta, citrate synthase and acetyl-CoA acetyltransferase. Immunoblot detection using chemiluminiscence showed possible nitration of other proteins such as cytochrome b-c1 complex subunit 1, NADH dehydrogenase [ubiquinone] ironsulfur protein 2, elongation factor Tu, NADH dehydrogenase [ubiquinone] flavoprotein 2, heat shock protein beta-1 and NADH dehydrogenase [ubiquinone] iron-sulfur protein 8. ATP synthase beta subunit was nitrated both in membrane and in fraction prepared by osmotic lysis. The high sensitivity of proteins to nitration by peroxynitrite is of potential biological importance, as these enzymes are involved in various pathways associated with energy production in the heart. [ABSTRACT FROM AUTHOR]

Arthrospira platensis is a species of filamentous cyanobacteria whose high protein content renders it suitable as a nutritive supplement for human and animal diets. Furthermore it is easy to grow as it tolerates a broad range of environmental stresses such as extremes of pH, salinity and heavy metals. In this study, changes in the growth, pigments and proteome of A. platensis were analyzed under two major abiotic stresses limiting productivity, salinity and iron. The lower concentrations of stresses showed varied effects on fresh weight, growth rate, pigment constituents and soluble protein of A. platensis . A proteomic analysis was conducted using SDS-PAGE first to obtain an overview of gross changes, followed by 2-DE and mass spectrometry to identify specific proteins whose abundance was affected by these environmental stresses. Eighteen spots were differentially expressed under the stress conditions. Of these, six were found with increased abundance responding only to Fe 2+ stress, and five as a result of NaCl stress alone. Using LC–MS/MS, 37 proteins were identified within these spots. These proteins were classified into thirteen categories according to their function annotation. The present analysis could provide important data related to stress-tolerance by microalgae. [ABSTRACT FROM AUTHOR]

Contents: Parturition is one of the most important events in reproduction. Regardless of many studies, exact time for pregnancy termination and onset of parturition is impossible to determine. The aim of this study was to describe and to compare protein profile of plasma from healthy pregnant cows (n = 6) at following five time points: 2 weeks, 1 week before, at parturition, 1 week and 2 weeks after parturition to search for possible protein markers of parturition. Plasma samples were analysed by 1D and 2D electrophoresis, and selected spots were identified by mass spectrometry. Protein profile showed no uniform pattern. Seventy spots differed at least for one sampling point from the time point 2 weeks before parturition which served as reference. Thirty spots expressed higher intensity of staining 1 week as 2 weeks before parturition while 13 showed opposite relationship. Twenty two spots expressed higher intensity of staining at parturition as 2 weeks before delivery while 15 showed opposite relationship. Eighteen spots expressed higher intensity of staining 2 weeks before parturition as 1 week post‐partum while 2 showed opposite relationship. Fifteen spots expressed higher intensity of staining 2 weeks before parturition as 2 weeks after delivery while 14 showed opposite relationship. Thirty‐five proteins, belonging to different functional groups, were identified. Of them, 15 spots differed significantly between parturition and 2 weeks before delivery. Among them were metalloproteinase inhibitor and LDH which seem to be the most promising molecules considered as parturition markers due to their functions. [ABSTRACT FROM AUTHOR]

Many clinically available anticancer compounds are designed to target DNA. This commonality of action often yields overlapping cellular response mechanisms and can thus detract from drug efficacy. New compounds are required to overcome resistance mechanisms that effectively neutralise compounds like cisplatin and those with similar chemical structures. Studies have shown that 56MESS is a novel compound which, unlike cisplatin, does not covalently bind to DNA, but is more toxic to many cell lines and active against cisplatin-resistant cells. Furthermore, a transcriptional study of 56MESS in yeast has implicated iron and copper metabolism as well as the general yeast stress response following challenge with 56MESS. Beyond this, the cytotoxicity of 56MESS remains largely uncharacterised. Here, yeast was used as a model system to facilitate a systems-level comparison between 56MESS and cisplatin. Preliminary experiments indicated that higher concentrations than seen in similar studies be used. Although a DNA interaction with 56MESS had been theorized, this work indicated that an effect on protein synthesis/ degradation was also implicated in the mechanism(s) of action of this novel anticancer compound. In contrast to cisplatin, the different mechanisms of action that are indicated for 56MESS suggest that this compound could overcome cisplatin resistance either as a stand-alone treatment or a synergistic component of therapeutics.

Crohn's disease (CD) represents a highly debilitating disease of difficult diagnosis and increasing incidence. Serum protein profiling of early stage Crohn's disease (ES) CD was investigated in order to improve the comprehension of the very early pathologic mechanisms and to support the difficult diagnostic procedures currently available. Inflammatory proteins and complement 3 chain C (C3c) were over-represented during ES CD, clusterin, retinol binding protein, α1-microglobulin and transthyretin were under-represented. A C3c isoform was found to be present only during ES CD. By now, lack of specific antibodies to detect isoforms made it impossible to perform alternative validation.

Background Bird fancier's lung (BFL) caused by repeated inhalation of avian proteins is the most common form of hypersensitivity pneumonitis. However, the exact identification of proteins involved is unknown, and serological test use for diagnosis need to be standardized. The objectives of this study were (i) to identify antigenic proteins from pigeon droppings (ii) to provide information about their location in avian matrices and (iii) to produce them in recombinant proteins to evaluate their diagnostic performances. Method Antigenic proteins of pigeon dropping extracts were investigated using 2-dimensional immunoblotting with sera from patients with BFL, asymptomatic exposed controls and healthy volunteers. We investigated the origin of these antigenic proteins by analyzing droppings, blooms and sera using a shotgun proteomic analysis. BFL-associated proteins were produced as recombinant antigens in E. coli and were assessed in ELISA with sera from patients (n = 25) and subject exposed controls (n = 30). These diagnostic performances were compared with those obtained by precipitin techniques (agar gel double diffusion, immunoelectrophoresis). Results We identified 14 antigenic proteins mainly located in droppings and blooms. These proteins were involved in either the digestive or immune systems of pigeons. Using the recombinant BFL-associated proteins: Immunoglobulin lambda-like polypeptide-1 (IGLL1: sensitivity: 76%; specificity: 100%; AUC: 0.93) and Proproteinase E (ProE: sensitivity: 84%; specificity: 80%; AUC: 0.85), the ELISA test showed better performance than precipitin assays with pigeon dropping extracts (sensitivity: 60%; specificity: 93.3%; AUC: 0.76). Conclusion IGLL1 and ProE were identified as the biomarkers of the disease. The use of these highly standardized antigens discriminates BFL cases from exposed subjects in serological assays. The results of this study offer new possibilities for the serological diagnosis of the disease. Clinical trial registration: ClinicalTrials.gov : Identifier NCT03056404 . [ABSTRACT FROM AUTHOR]