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Many foods including edible oils contain 2-monochloropropane-1,3-diol (2-MCPD), a processing-induced chemical contaminant. Cardiotoxic effects have been shown to result from oral 2-MCPD exposure in rodents, but the underlying mechanisms of action remain poorly understood. We undertook a comprehensive multi-omics approach to assess changes at the transcriptomic, proteomic, and oxylipin levels in heart tissues from male F344 rats that were exposed to 0 or 40 mg/kg BW/day of 2-MCPD in the diet for 90 days, in a regulatory compliant rodent bioassay. Heart tissues were collected for RNA sequencing, quantitative PCR analysis, proteomic analysis via two-dimensional gel electrophoresis and mass spectrometry, and targeted lipidomic profiling by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). Transcriptomic and proteomic data analyses revealed upregulation of immune/inflammatory response processes and downregulation of energy metabolism and cardiac structure and functions. Among differentially expressed gene–protein pairs, coronin-1A, a key leukocyte-regulating protein, emerged as markedly up-regulated. Oxylipin profiling highlighted a selective suppression of docosahexaenoic acid-derived metabolites, suggesting a disruption in cardioprotective lipid pathways. These findings suggest that 2-MCPD disrupts homeostasis through inflammatory activation and suppression of metabolic and cardiac function. This research provides insights into 2-MCPD's cardiotoxicity, emphasizing the need for further studies to support hazard characterization. [ABSTRACT FROM AUTHOR]

Inhalation of ambient particulate matter (PM) and ozone (O3) has been associated with increased cardiovascular morbidity and mortality. However, the interactive effects of PM and O3 on cardiac dysfunction and disease have not been thoroughly examined, especially at a proteomic level. The purpose of this study was to identify and compare proteome changes in spontaneously hypertensive (SH) rats co-exposed to concentrated ambient particulates (CAPs) and O3, with a focus on investigating inflammatory and metabolic pathways, which are the two major ones implicated in the pathophysiology of cardiac dysfunction. For this, we measured and compared changes in expression status of 9 critical pro- and anti-inflammatory cytokines using multiplexed ELISA and 450 metabolic proteins involved in ATP production, oxidative phosphorylation, cytoskeletal organization, and stress response using two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) in cardiac tissue of SH rats exposed to CAPs alone, O3 alone, and CAPs + O3. Proteomic expression profiling revealed that CAPs alone, O3 alone, and CAPs + O3 differentially altered protein expression patterns, and utilized divergent mechanisms to affect inflammatory and metabolic pathways and responses. Ingenuity Pathway Analysis (IPA) of the proteomic data demonstrated that the metabolic protein network centered by gap junction alpha-1 protein (GJA 1) was interconnected with the inflammatory cytokine network centered by nuclear factor kappa beta (NF-kB) potentially suggesting inflammation-induced alterations in metabolic pathways, or vice versa, collectively contributing to the development of cardiac dysfunction in response to CAPs and O3 exposure. These findings may enhance understanding of the pathophysiology of cardiac dysfunction induced by air pollution and provide testable hypotheses regarding mechanisms of action. [ABSTRACT FROM AUTHOR]

In this study, the optimal hydrogen (H 2) production conditions of the high-efficiency H 2 -producing mutant strain Ethanoligenens harbinense YR-3 (carbon-nitrogen ratio 5.5, phosphate buffer 80 mM, initial pH 6.0, biotin 1.4 mg/L) are obtained by intermittent experiments. The maximum specific H 2 production rate of YR-3 (2.85 mol H 2 /mol glucose) was 1.4 times that of the wild strain ZGX4 (2.04 mol H 2 /mol glucose). The liquid-phase products are mainly ethanol and acetic acid, indicating that the metabolic pathway has not changed. Two-dimensional electrophoresis and mass spectrometry were used to compare and analyze the protein map differences between YR-3 and ZGX4. The results show that 1,6-fructose diphosphate aldolase and the flavoprotein in hydrogenase are highly expressed. This study will provide a theoretical basis for the genetic modification of high-efficiency H 2 -producing strains and the improvement of H 2 production capacity. [ABSTRACT FROM AUTHOR]

This study investigates the changes in hippocampal proteomic profiles during demyelination and remyelination using the cuprizone model. Employing two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry for protein profiling, we observed significant alterations in the expression of ketimine reductase mu-crystallin (CRYM) and protein disulfide isomerase A3 precursor (PDIA3) following exposure to and subsequent withdrawal from cuprizone. Immunohistochemical staining validated these protein expression patterns in the hippocampus, revealing that both PDIA3 and CRYM were downregulated in the hippocampal CA1 region during demyelination and upregulated during remyelination. Additionally, we explored the potential protective effects of CRYM and PDIA3 against cuprizone-induced demyelination by synthesizing cell-permeable Tat peptide-fusion proteins (Tat-CRYM and Tat-PDIA3) to facilitate their crossing through the blood–brain barrier. Our results indicated that administering Tat-CRYM and Tat-PDIA3 mitigated the reduction in proliferating cell and differentiated neuroblast counts compared to the group receiving cuprizone alone. Notably, Tat-PDIA3 demonstrated significant effects in enhancing myelin basic protein expression alongside phosphorylation of CREB in the hippocampus, suggesting its potential therapeutic role in the prevention or treatment of demyelination, and by extension, in conditions such as multiple sclerosis. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department 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.)

The polyketide antibiotic 2,4-diacetylphloroglucinol (DAPG) is a phenolic secondary metabolite produced by Pseudomonas spp. that serves as a biocontrol agent. DAPG is known to inhibit root growth and seed germination in plants. However, the effects of DAPG on leaf growth have not been elucidated yet. In this study, we found that applying DAPG at concentration above 30 µM suppresses the development of Arabidopsis leaves. Using an integrated proteomic analysis approach, we investigated the physio-molecular responses of Arabidopsis leaves under 10–100 µM DAPG treatment. Two-dimensional electrophoresis (2-DE)-based proteomic analysis identified 46 differentially expressed spots with at least a twofold change, corresponding to 37 proteins involved in stress and defense, metabolism, photosynthesis, and carbohydrate metabolism processes. Furthermore, we found that the expressions of stress and defense-related proteins were upregulated in response to DAPG, while those of photosynthesis-related proteins were downregulated. Additionally, RT-PCR result showed that the identified DAPG-responsive proteins in the proteomic analysis were similarly regulated at the transcriptional level. Collectively, our findings provide insights into the molecular mechanism of the response to DAPG in Arabidopsis leaves. [ABSTRACT FROM AUTHOR]

Amaranth is a promising staple food that produces seeds with excellent nutritional quality. Although cultivated species intended for grain production have interesting agronomic traits, relatively little is known about wild species, which can prosper in diverse environments and could be a rich genetic source for crop improvement. This work focuses on the proteomic comparison between the seeds of wild and cultivated amaranth species using polarity-based protein extraction and two-dimensional gel electrophoresis. Differentially accumulated proteins (DAPs) showed changes in granule-bound starch synthases and a wide range of 11S globulin isoforms. The electrophoretic profile of these proteins suggests that they may contain significant phosphorylation as post-translational modifications (PTMs), which were confirmed via immunodetection. These PTMs may impact the physicochemical functionality of storage proteins, with potential implications for seed agronomic traits and food system applications. Low-abundant DAPs with highly variable accumulation patterns are also discussed; these were involved in diverse molecular processes, such as genic regulation, lipid storage, and stress response. [ABSTRACT FROM AUTHOR]

Acacia mearnsii De Wild (black wattle) is one of the most important forestry crops in South Africa. The bark of this species contains high-quality tannins, while its wood is used for wood chips, charcoal and mining timber. One of the main problems faced by black wattle farmers is frost damage, thus there is an urgent need to develop frost-tolerant black wattle varieties. A quick method for the screening of frost tolerance variation would be beneficial for black wattle breeders and would result in the need for fewer families to undergo field trials. Frost tolerance in plants is directly related to their ability to undergo cold acclimation in response to changing weather conditions. During the cold acclimation process, many physiological changes occur, including modifications in protein expression. In this study, two-dimensional gel electrophoresis was used to investigate changes in protein expression among 40 black wattle families with varying levels of frost tolerance before and after a cold-stress treatment. The proteomic data generated from this was analysed using multivariate analysis. Six proteins were observed to be upregulated in frosttolerant black wattle families. The proteins were identified and have been previously shown to contribute to the protection of cellular membranes, maintenance of photosynthetic processes and the prevention of protein misfolding and aggregation. [ABSTRACT FROM AUTHOR]

Some health disorders, such as obesity and type 2 diabetes, are associated with a poor diet and low quality of the fat in it. The type and duration of the diet have an impact on the liver. This investigation uses the proteomic approach to identify changes in the mouse liver protein profile in adaptation to high-fat diets with different saturated fatty acid contents and linoleic acid (18:2n-6) to α-linolenic acid (18:3n-3) fatty acid ratios.

The recent advances in pigeon pea genomics, including high-quality whole genome and chloroplast genome sequence information helped develop improved varieties. However, a comprehensive Cajanus proteome, including the organelle proteome, is yet to be fully mapped. The spatial delineation of pigeon pea proteins at sub-cellular levels and inter-organelle communication could offer valuable insights into its defense mechanism against various stresses. However, the major bottleneck in the proteomic study is the lack of a suitable method of protein extraction and sample preparation compatible with two-dimensional gel electrophoresis (2D-PAGE), liquid chromatography-mass spectrometry (LCMS), or matrix-assisted laser desorption ionization-time of flight (MALDi-ToF). Our study introduces two efficient methods, one for isolating total proteins and another for organelle (chloroplast) proteins from various Cajanus spp. For total protein extraction, we have optimized a protocol using phenol in combination with a reducing agent (DTT) and protease inhibitor cocktail, also washing (6–7 times) with ice-cold acetone after overnight protein precipitation of total proteins. Our modified extraction method using phenol for total leaf protein yielded approximately 2-fold more proteins than the previously reported protocols from C. cajan (3.18 ± 0.11 mg/gm) and C. scarabaeoides (2.06 ± 0.08 mg/gm). We have also optimized a protocol for plastid protein extraction, which yielded 1.33 ± 0.25 mg/10 gm plastid proteins from C. cajan and 0.88 ± 0.19 mg/10 gm plastid proteins from C. scarabaeoides. The 2D-PAGE analysis revealed 678 ± 08 reproducible total protein spots from C. cajan and 597 ± 22 protein spots from C. scarabaeoides. Similarly, we found 566 ± 10 and 486 ± 14 reproducible chloroplast protein spots in C. cajan and C. scarabaeoides, respectively. We confirmed the plastid protein fractions through immunoblot analysis using antibodies against LHCb1/LHCⅡ type Ⅰ protein. We found both methods suitable for 2D-PAGE and mass spectrometry (MS). This is the first report on developing protocols for total and chloroplastic protein extraction of Cajanus spp. suitable for advanced proteomics research. [ABSTRACT FROM AUTHOR]

Viral diseases of potatoes are among the main problems causing deterioration in the quality of tubers and loss of yield. The growth and development of potato plants largely depend on soil moisture. Prevention strategies require comprehensive protection against pathogens and abiotic stresses, including modeling the beneficial microbiome of agroecosystems combining microorganisms and immunostimulants. Chitosan and its derivatives have great potential for use in agricultural engineering due to their ability to induce plant immune responses. The effect of chitosan conjugate with caffeic acid (ChCA) in combination with Bacillus subtilis 47 on the transcriptional activity of PR protein genes and changes in the proteome of potato plants during potato virus Y (PVY) infection and drought was studied. The mechanisms of increasing the resistance of potato plants to PVY and lack of moisture are associated with the activation of transcription of genes encoding PR proteins: the main protective protein (PR-1), chitinase (PR-3), thaumatin-like protein (PR-5), protease inhibitor (PR-6), peroxidase (PR-9), and ribonuclease (PR-10), as well as qualitative and quantitative changes in the plant proteome. The revealed activation of the expression of marker genes of systemic acquired resistance and induced systemic resistance under the influence of combined treatment with B. subtilis and chitosan conjugate indicate that, in potato plants, the formation of resistance to viral infection in drought conditions proceeds synergistically. By two-dimensional electrophoresis of S. tuberosum leaf proteins followed by MALDI-TOF analysis, 10 proteins were identified, the content and composition of which differed depending on the experiment variant. In infected plants treated with ChCA, the synthesis of proteinaceous RNase P 1 and oxygen-evolving enhancer protein 2 was enhanced in conditions of normal humidity, and 20 kDa chaperonin and TMV resistance protein N-like was enhanced in conditions of lack of moisture. The virus coat proteins were detected, which intensively accumulated in the leaves of plants infected with potato Y-virus. ChCA treatment reduced the content of these proteins in the leaves, and in plants treated with ChCA in combination with Bacillus subtilis, viral proteins were not detected at all, both in conditions of normal humidity and lack of moisture, which suggests the promising use of chitosan derivatives in combination with B. subtilis bacteria in the regulation of plant resistance. [ABSTRACT FROM AUTHOR]

Although helminth parasites have different life cycles, their hosts share similar immune responses involving Th2 cell-type. Here, we extracted proteins from the larvae of Anisakis simplex complex and Trichinella spiralis to identify common and specific antigens (or allergens) associated with the Th2 immune response. We performed two-dimensional electrophoresis analysis and Matrix-assisted laser desorption ionization–time of flight/time of flight (MALDI-TOF/TOF) experiments. We found 13 potentially immunogenic proteins, which included 5 spots specific to T. spiralis and 8 common to T. spiralis and A. simplex, by tandem mass spectrometry. These molecules were identified structurally as actin, tropomyosin, col cuticle N domain-containing protein, and heat shock proteins. We also identified molecules related to parasite-host immune modulation and interactions. Our results may contribute to reveal potential roles of immunological proteins in parasite-derived immune modulation. [ABSTRACT FROM AUTHOR]

Pregnancy has its own protein dynamics, reflecting the hormonal profile. Quantitative and qualitative changes in plasma protein profile may provide useful information about this condition. Any alterations may be a signal heralding clinical or subclinical pathology. The objective of our study was to compare the plasma protein profile between selected months of pregnancy in cows for a better understanding gestation course. For this purpose, we collected blood from healthy pregnant (n = 30; n = 6 for each pregnancy stage) and non‐pregnant (C; n = 6) Holstein‐Friesian cows during a routine veterinary examination. Collected samples were selected according to pregnancy month (first, second, third, sixth, and ninth), prepared, and separated by two‐dimensional electrophoresis. The Delta‐2D program compared and statistically evaluated scanned gel images from the appropriate months. The mean volume of the spots was considered. The MALDI TOF/TOF spectrometer was used to identify statistically significant proteins. There were 11 distinct proteins found, including peptidyl‐prolyl cis‐trans isomerase F, oligoribonuclease, and PRELI domain‐containing protein 3B (all of them have the lowest abundance in the C group), alpha‐1B‐glycoprotein, L‐gulonolactone oxidase, hemopexin (first month with higher abundance than control), alpha‐2‐HS‐glycoprotein (significantly higher abundance in the first month than in remaining groups), ermin (absent in the first month and lower abundance in the third and sixth months than in the remaining groups and control), endophilin‐A2 (significant differences between the control and the second, third, sixth, and ninth months), apolipoprotein A‐I (significant difference between control and the first and sixth months), alpha‐1‐antiproteinase (significant difference between control and the ninth month). The study demonstrated the distinctions between plasma protein composition and alterations during the pregnancy course which may potentially serve as diagnostic tools. [ABSTRACT FROM AUTHOR]

Two‐dimensional electrophoresis (2DE) is a gel‐based protein separation method based on size and charge which is commonly used for the characterization of host cell proteins (HCPs) during drug development in biotech and pharmaceutical companies. HCPs are a heterogenous mixture of proteins produced by host cells during a biologics drug manufacturing process. Different gel electrophoresis methods including traditional 2D SDS‐PAGE with silver and SYPRO Ruby fluorescent dye staining as well as two‐dimensional difference gel electrophoresis (2D‐DIGE) were compared for their relative abilities to characterize HCPs. SYPRO Ruby was shown to be more sensitive than silver stain in the traditional 2D gels both with and without product protein present. Silver stain also displayed a significant preference for staining acidic proteins over basic ones while SYPRO Ruby was more consistent in imaging proteins across different isoelectric points. The non‐traditional method of 2D‐DIGE provides high resolution and reproducibility when comparing samples with similar protein profiles but was limited in imaging HCP spots due to its narrow dynamic range. Overall, 2DE is a powerful tool to separate and characterize HCPs and is optimized by choosing the best stain or method for each specific application. Using a combination of two or more different 2DE staining methods, when possible, provides the most comprehensive coverage to support the characterization of a complex mixture like HCPs. However, in instances where only one staining method can be used, SYPRO Ruby is shown to be the more reliable, more sensitive, and easier to use traditional staining method for most HCP‐based applications. [ABSTRACT FROM AUTHOR]

Toxoplasmosis, while often asymptomatic and prevalent as a foodborne disease, poses a considerable mortality risk for immunocompromised individuals during pregnancy. Point-of-care serological tests that detect specific IgG and IgM in patient sera are critical for disease management under limited resources. Despite many efforts to replace the T. gondii total lysate antigens (TLAs) by recombinant antigens (rAgs) in commercial kits, while IgG detection provides significant specificity and sensitivity, IgM detection remains comparatively low in sensitivity. In this study, we attempted to identify novel antigens targeting IgM in early infection, thereby establishing an IgM on-site detection kit. Using two-dimensional gel electrophoresis (2DE) and mouse serum immunoblotting, three novel antigens, including EF1γ, PGKI, and GAP50, were indicated to target T. gondii IgM. However, rAg EF1γ was undetectable by IgM of mice sera in Western blotting verification experiments, and ELISA coated with PGKI did not eliminate cross-reactivity, in contrast to GAP50. Subsequently, the lateral flow reaction employing a strip coated with 0.3 mg/mL purified rAg GAP50 and exhibited remarkable sensitivity compared with the conventional ELISA based on tachyzoite TLA, which successfully identified IgM in mouse sera infected with tachyzoites, ranging from 10³ to 104 at 5 dpi and 104 at 7 dpi, respectively. Furthermore, by using standard T. gondii-infected human sera from WHO, the limit of detection (LOD) for the rapid fluorescence immunochromatographic test (FICT) using GAP50 was observed at 0.65 IU (international unit). These findings underline the particular immunoreactivity of GAP50, suggesting its potential as a specific biomarker for increasing the sensitivity of the FICT in IgM detection. [ABSTRACT FROM AUTHOR]

Aims Acinetobacter baumannii is a nosocomial pathogen known to be multidrug-resistant (MDR), especially to drugs of the carbapenem class. Several factors contribute to resistance, including efflux pumps, β -lactamases, alteration of target sites, and permeability defects. In addition, outer membrane proteins (OMPs), like porins are involved in the passage of antibiotics, and their alteration could lead to resistance development. This study aimed to explore the possible involvement of porins and OMPs in developing carbapenem resistance due to differential expression. Methods and results The antibiotic-susceptible and MDR isolates of A. baumannii were first studied for differences in their transcriptional levels of OMP expression and OMP profiles. The antibiotic-susceptible isolates were further treated with imipenem, and it was found that the omp genes were differentially expressed. Six of the nine genes studied were upregulated at 1 h of exposure to imipenem. Their expression gradually decreased with time, further confirmed by their OMP profile and two-dimensional gel electrophoresis. Conclusions This study could identify OMPs that were differentially expressed on exposure to imipenem. Hence, this study provides insights into the role of specific OMPs in antibiotic resistance in A. baumannii. [ABSTRACT FROM AUTHOR]

Plants initiate specific defense responses by recognizing conserved epitope peptides within the flagellin proteins derived from bacteria. Proteolytic cleavage of epitope peptides from flagellin by plant apoplastic proteases is thought to be crucial for the perception of the epitope by the plant receptor. However, the identity of the plant proteases involved in this process remains unknown. Here, we establish an efficient identification system for the target proteases in Arabidopsis apoplastic fluid; the method employs native two-dimensional electrophoresis followed by an in-gel proteolytic assay using a fluorescence-quenching peptide substrate. We designed a substrate to specifically detect proteolytic activity at the C-terminus of the flg22 epitope in flagellin and identified two plant subtilases, SBT5.2 and SBT1.7, as specific proteases responsible for the C-terminal cleavage of flg22. In the apoplastic fluid of Arabidopsis mutant plants deficient in these two proteases, we observe a decrease in the C-terminal cleavage of the flg22 domain from flagellin, leading to a decrease in the efficiency of flg22 epitope liberation. Consequently, defensive reactive oxygen species (ROS) production is delayed in sbt5.2 sbt1.7 double-mutant leaf disks compared to wild type following flagellin exposure. Plants initiate specific defense responses by recognizing flg22 peptide epitope derived from flagellin, the major structural protein of the bacterial flagellum. Here, the authors identified two plant subtilases, SBT5.2 and SBT1.7, as specific proteases responsible for cleaving the peptide epitope. [ABSTRACT FROM AUTHOR]

Copyright of Journal of China Medical University is the property of Journal of China Medical University Editorial Office 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.)

Two-dimensional electrophoresis (2-DE) is an important method for studying urinary proteomics, playing an important role in assessing changes in protein concentrations under different physiological conditions. This study aimed to evaluate the different proteins expressed in the urine of normal cycling, healthy cows. We attained the images from 2-DE of urinary proteins in cow urine. Then, we implemented and evaluated an image analysis protocol with 2D ImageMaster software. Using ImageMaster, different spots were identified. On two-dimensional electrophoresis gels, 40 protein spots from anionic fractions and 15 spots from the cationic fractions of urine were identified. These spots corresponded to a molecular weight (MW) of 10-113 kDa for anionic fractions, and a MW of 15-51 kDa in cationic fractions. There were 12 master spot features across both the gels. This dataset can serve as a foundation for further investigations into the functional roles of specific proteins and their implications in the biological system under study. [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the proteins in canine seminal plasma with direct effect on specific sperm parameters during fertilisation. The influence of seminal plasma proteins was fo-cused on three processes: capacitation, hyperactivation and decapacitation. Size exclusion chroma-tographic fractional separation of proteins from seminal plasma based on their molecular weights was performed. In addition, two-dimensional electrophoresis of two seminal protein fractions was done. The effect of protein molecules on the motion and kinetic parameters of the male gametes in in vitro conditions was monitored by computer-assisted sperm analysis. The activity of alkaline phosphatase, creatine kinase and alanine aminotransferase in whole seminal plasma and each chromatography fraction was measured spectrophotometrically using the BA-88A semi-automatic chemistry analyzer. The protein fractions with different molecular weights affected the in vitro motility and kinetic characteristics (curvilinear velocity, straightness, linearity and amplitude of lateral head displacement) of spermatozoa, leading to hyperactivation or initiating decapacitation kinetics changes. The detailed study of the seminal plasma proteome would add fundamental information about the processes associated with physiological changes occurring in spermatozoa before fertilisation. The study of the canine seminal plasma proteome could add relevant information about its effects on the fertilising ability of the male gametes and the changes occurring in them before fertilisation.

The aim of the present study was to determine the proteins in canine seminal plasma with direct effect on specific sperm parameters during fertilisation. The influence of seminal plasma proteins was focused on three processes: capacitation, hyperactivation and decapacitation. Size exclusion chromatographic fractional separation of proteins from seminal plasma based on their molecular weights was performed. In addition, two-dimensional electrophoresis of two seminal protein fractions was done. The effect of protein molecules on the motion and kinetic parameters of the male gametes in in vitro conditions was monitored by computer-assisted sperm analysis. The activity of alkaline phosphatase, creatine kinase and alanine aminotransferase in whole seminal plasma and each chromatography fraction was measured spectrophotometrically using the BA-88A semi-automatic chemistry analyzer. The protein fractions with different molecular weights affected the in vitro motility and kinetic characteristics (curvilinear velocity, straightness, linearity and amplitude of lateral head displacement) of spermatozoa, leading to hyperactivation or initiating decapacitation kinetics changes. The detailed study of the seminal plasma proteome would add fundamental information about the processes associated with physiological changes occurring in spermatozoa before fertilisation. The study of the canine seminal plasma proteome could add relevant information about its effects on the fertilising ability of the male gametes and the changes occurring in them before fertilisation. [ABSTRACT FROM AUTHOR]

Purpose: Visceral leishmaniasis (VL) is a systemic and parasitic disease that is usually fatal if left untreated. VL is endemic in different parts of Iran and is caused mainly by Leishmania infantum. This study aimed to recognition immunoreactive proteins in amastigote-like and promastigote stages of L. infantum (Iranian strain) by antibodies present in the sera of VL patients. Methods: Total protein extract from amastigote-like and promastigote cells was separated by two-dimensional electrophoresis (2DE). To detect the immunoreactive proteins, 2DE immunoblotting method was performed using different pools of VL patients' sera. Results: Approximately 390 and 430 protein spots could be separated in 2DE profiles of L. infantum amastigote-like and promastigote stages, respectively. In immunoblotting method, approximately 295 and 135 immunoreactive proteins of amastigotes-like reacted with high antibody titer serum pool and low antibody titer serum pool, respectively. Approximately 120 and 85 immunoreactive proteins of promastigote extract were recognized using the high antibody titer sera pool and low antibody titer sera, respectively. Conclusion: The present study has recognized a number of antigenic diversity proteins based on the molecular weight and pH in amastigote-like and promastigote stages of L. infantum. These results provide us a new concept for further analysis development in the field of diagnosis biomarkers and vaccine targets. [ABSTRACT FROM AUTHOR]

Results obtained from rat studies indicate that, even at low concentrations, mercurial species cause harmful effects on the kidneys, by inducing the nephrotic oxidative stress response. In the present work, Hg-associated proteins were identified as possible mercury-exposure biomarkers in rat kidneys exposed to low mercury chloride concentrations for 30 days (Hg-30) and 60 days (Hg-60), using metalloproteomic strategies. The renal proteomic profile was fractioned by two-dimensional electrophoresis and the mercury determinations in kidney samples, protein pellets and protein spots were performed using graphite furnace atomic absorption spectrometry. The characterization of Hg-associated protein spots and the analysis of differentially expressed proteins were performed by liquid chromatography, coupled with tandem mass spectrometry. Eleven Hg-associated protein spots with a concentration range of 79 ± 1 to 750 ± 9 mg kg−1 in the Hg-60 group were identified. The characterization and expression analyses allowed the identification of 53 proteins that were expressed only in the Hg-60 group, 13 "upregulated" proteins (p > 0.95) and 47 "downregulated" proteins (p < 0.05). Actin isoforms and hemoglobin subunits were identified in protein spots of the Hg-60 group, with mercury concentrations in the range of 138 to 750 mg kg−1, which qualifies these proteins as potential mercury-exposure biomarkers. [ABSTRACT FROM AUTHOR]

Boron, in the form of potassium tetraborate, has previously been found to be effective at inhibiting mango black spot disease, caused by Alternaria alternata. However, the mechanisms involved in this inhibition are largely unknown. In this study, A. alternata was treated in vitro with potassium tetraborate at a concentration of 5–10 mM for 48 or 72 h. The intracellular and mitochondrial proteins were extracted from mycelium and separated using two‐dimensional electrophoresis (2‐DE). Differentially expressed proteins (DEPs) were identified using bioinformatics tools and differences between protein spots were derived from mass spectrometry (MS). Using matrix‐assisted laser desorption ionization time‐of‐flight tandem mass spectrometry (MALDI‐ToF‐MS/MS), 96 intracellular and 56 mitochondrial DEPs were identified. The intracellular proteins identified were found to be involved in posttranslational modifications, protein turnover and chaperones, while the mitochondrial proteins were involved in electron transport chains. Our results demonstrate that various metabolic pathways are involved in the antifungal activity of boron. The differential expression of 20 genes was also verified at the mRNA level by reverse transcription‐quantitative PCR. Our study suggests that borate could be used as a potential substitute for synthetic fungicides to control this postharvest disease of mango fruits. [ABSTRACT FROM AUTHOR]

Phaeocystis globosa is an important unicellular eukaryotic alga that can also form colonies. P. globosa can cause massive harmful algal blooms and plays an important role in the global carbon or sulfur cycling. Thus far, the ecophysiology of P. globosa has been investigated by numerous studies. However, the proteomic response of P. globosa to nitrogen depletion remains largely unknown. We compared four protein preparation methods of P. globosa for two-dimensional electrophoresis (2-DE) (Urea/Triton X-100 with trichloroacetic acid (TCA)/acetone precipitation; TCA/acetone precipitation; Radio Immuno Precipitation Assay (RIPA) with TCA/acetone precipitation; and Tris buffer). Results show that the combination of RIPA with TCA/acetone precipitation had a clear gel background and showed the best protein spot separation effect, based on which the proteomic response to nitrogen depletion was studied using 2-DE. In addition, we identified six differentially expressed proteins whose relative abundance increased or decreased more than 1.5-fold (P<0.05). Most proteins could not be identified, which might be attributed to the lack of genomic sequences of P. globosa. Under nitrogen limitation, replication protein-like, RNA ligase, and sn-glycerol-3-phosphate dehydrogenase were reduced, which may decrease the DNA replication level and ATP production in P. globosa cells. The increase of endonuclease III and transcriptional regulator enzyme may affect the metabolic and antioxidant function of P. globosa cells and induce cell apoptosis. These findings provide a basis for further proteomic study of P. globosa and the optimization of protein preparation methods of marine microalgae. [ABSTRACT FROM AUTHOR]

The resistance of silkworms to Bombyx mori nuclear polyhedrosis virus (BmNPV) is controlled by a major dominant gene and multiple modifying genes. Given the presence of modified genes, it is difficult to determine the main gene by positional cloning. In this study, the main anti-BmNPV gene of BmNPV-resistant silkworm variety N was introduced into the susceptible variety Su to breed the near-isogenic line SuN with BmNPV resistance. The infection process of BmNPV in the hemolymph of Su and SuN was analyzed using the cell analysis system TissueFAXS PLUS. According to the law of infection and proliferation, hemolymph was extracted every 6 h for two-dimensional electrophoresis (2-DE) analysis and quantitative real-time polymerase chain reaction (qRT-PCR). Seven DEPs were found in comparisons between Su and SuN by 2-DE analysis. Among them, acid phosphatase, storage protein, and phenoloxidase can prevent pathogen invasion, which may play a role against BmNPV. Polyamine oxidase plays an important role in energy metabolism, which may be indirectly involved in the process of resisting BmNPV. Most of the transcriptional expression profiles of the seven DEPs were consistent with the 2-DE results. This study can provide a reference for the identification of anti-BmNPV genes and the breeding of BmNPV-resistant silkworm varieties. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to investigate the presence of autoantigens in the gastric juices of children. Methods: Gastric juice and serum samples were obtained from 53 children <15 years of age who underwent gastric endoscopy. Among these, 8, 22, and 23 participants were in the age groups 0-5, 6-10, and 11-15 years, respectively. These samples were analyzed using two-dimensional electrophoresis (2-DE), immunoblot analysis, and matrix-assisted laser desorption ionization-time of-flight mass spectrometry. Furthermore, we reviewed the histopathological findings and urease test results and compared them with the results of 2-DE and immunoblot analysis. Results: There were no statistically significant differences in urease test positivity, grades of chronic gastritis, active gastritis, or Helicobacter pylori infiltration of the antrum and body among the three age groups. Three distinct patterns of gastric juice were observed on 2-DE. Pattern I was the most common, and pattern III was not observed below the age of 5 years. Histopathological findings were significantly different among active gastritis (p=0.037) and H. pylori infiltration (p=0.060) in the gastric body. The immunoblots showed large spots at an approximate pH of 3-4 and molecular weights of 31-45 kDa. These distinct, large positive spots were identified as gastric lipase and pepsin A and C. Conclusion: Three enzymes, which are normally secreted under acidic conditions were identified as autoantigens. Further investigation of the pathophysiology and function of autoantigens in the stomach is required. [ABSTRACT FROM AUTHOR]

Forkhead box E1 (FoxE1) protein is a transcriptional regulator known to play a major role in the development of the thyroid gland. By performing sequence alignments, we detected a deletion in FoxE1, which occurred in the evolution of mammals, near the point of divergence of placental mammals. This deletion led to the loss of the majority of the Eh1 motif, which was important for interactions with transcriptional corepressors. To investigate a potential mechanism for this deletion, we analyzed replication through the deletion area in mammalian cells with two-dimensional gel electrophoresis, and in vitro, using a primer extension reaction. We demonstrated that the area of the deletion presented an obstacle for replication in both assays. The exact position of polymerization arrest in primer extension indicated that it was most likely caused by a quadruplex DNA structure. The quadruplex structure hypothesis is also consistent with the exact borders of the deletion. The exact roles of these evolutionary changes in FoxE1 family proteins are still to be determined. [ABSTRACT FROM AUTHOR]

Adipose-derived mesenchymal stem cells (ASCs) have the potential to differentiate into bone, cartilage, fat, and neural cells and promote tissue regeneration and healing. It is known that they can have variable responses to hypoxic conditions. In the present study, we aimed to explore diverse changes in the cells and secretome of ASCs under a hypoxic environment over time and to present the possibility of ASCs as therapeutic agents from a different perspective. The expression differences of proteins between normoxic and hypoxic conditions (6, 12, or 24 h) were specifically investigated in human ASCs using 2-DE combined with MALDI-TOF MS analysis, and secreted proteins in ASC-derived conditioned media (ASC-derived CM) were examined by an adipokine array. In addition, genetic and/or proteomic interactions were assessed using a DAVID and miRNet functional annotation bioinformatics analysis. We found that 64 and 5 proteins were differentially expressed in hypoxic ASCs and in hypoxic ASC-derived CM, respectively. Moreover, 7 proteins among the 64 markedly changed spots in hypoxic ASCs were associated with bone-related diseases. We found that two proteins, cathepsin D (CTSD) and cathepsin L (CTSL), identified through an adipokine array independently exhibited significant efficacy in promoting osteocyte differentiation in bone-marrow-derived mesenchymal stem cells (BM-MSCs). This finding introduces a promising avenue for utilizing hypoxia-preconditioned ASC-derived CM as a potential therapeutic approach for bone-related diseases. [ABSTRACT FROM AUTHOR]

In the gastropod mollusk Lymnaea stagnalis, insulin-like peptides in the central nervous system (CNS) control behavioral changes associated with associative learning. Insulin administration to the Lymnaea CNS enhances the synaptic plasticity involved in this type of learning, but it has remained unclear which molecules in the insulin response cascade are involved. Here, to advance a comprehensive analysis, we used two-dimensional electrophoresis and comparative quantitative mass spectrometry to perform a protein analysis investigating the CNS molecules that respond to insulin administration. Our results revealed increased phosphorylation of AKT and RICTOR in the PI3K/AKT/mTOR signaling cascade and cytoskeleton-related proteins. Although it was expected that the molecules in the PI3K/AKT/mTOR signaling cascade were phosphorylated by insulin administration, our findings confirmed the correlation between insulin-induced phosphorylation of cytoskeletonrelated proteins strongly involved in the synaptic changes and learning and memory mechanisms. These results contribute to elucidate the relationship between the insulin response and learning and memory mechanisms not only in Lymnaea but also in various invertebrates and vertebrates. [ABSTRACT FROM AUTHOR]

Simple Summary: During the pupal stage, the midgut of Bombyx mori undergoes remarkable changes, condensing into the midgut lumen and, subsequently, undergoing decomposition. To comprehensively analyze this process of change, we utilized mass spectrometry to examine the midgut contents in the early and later stages of pupation. We detected a large number of enzymes involved in metabolism, proteases, inhibitors, chitin-binding proteins, chitinases, and storage proteins in the midgut contents during the pupal stage, indicating significant alterations within the pupal midgut lumen. Additionally, we have identified six proteases with potential gelatinolytic activity, which may be involved in the degradation of the midgut contents during the late stages of pupation. This research provides valuable insights into the mechanisms of tissue digestion and midgut remodeling in Bombyx mori pupae. The results serve as an important reference for comprehending the intricate processes taking place during this developmental stage in lepidopteran insects. The silkworm Bombyx mori, a lepidopteran insect, possesses an 8–10-day pupal stage, during which significant changes occur in the midgut, where it first condenses into the yellow body, and then undergoes decomposition. To gain insights into this transformation process, proteomics was performed on Bombyx mori midgut contents on day 2 and day 7 after pupation. The results revealed the identification of 771 proteins with more than one unique peptide. An analysis using AgriGO demonstrated that these proteins were predominantly associated with catalytic activity. Among the identified proteins, a considerable number were found to be involved in carbohydrate metabolism, amino acid metabolism, lipid metabolism, nucleic acid degradation, and energy support. Additionally, variations in the levels of certain proteases were observed between the midgut contents on day 2 and day 7 after pupation. An in-depth analysis of the two-dimensional electrophoresis of the midgut contents on day 7 after pupation led to the identification of twelve protein spots with potential gelatinolytic activity. Among these, six proteases were identified through mass spectrometry, including the p37k protease, vitellin-degrading protease, chymotrypsin-2, etc. These proteases may be responsible for the digestion of the yellow body during the later stages of pupal development. [ABSTRACT FROM AUTHOR]

Neutrophil adhesion to endothelia, entry into tissues and chemotaxis constitute essential steps in the immune response to infections that drive inflammation. Neutrophils bind to other cells and migrate via adhesion receptors, notably the αMβ2 integrin dimer (also called Mac-1, CR3 or CD11b/CD18). Here, the response of neutrophils to integrin engagement was examined by monitoring the activity of peptidylarginine deiminase 4 (PAD4). Histone H3 deimination was strongly stimulated by manganese, an integrin-activating divalent cation, even in the absence of additional inflammatory stimuli. Manganese-induced cell attachment resulted in neutrophil swarm formation that paralleled histone deimination, whereas antibodies that impair integrin binding prevented both cell adhesion and histone deimination. Manganese treatment led to putative deimination of profilin, a protein that functions as an actin-organizing hub, as detected by two-dimensional gel electrophoresis and citrulline immunoblotting. Cl-amidine, a covalent inhibitor of PAD4, and GSK484, a specific PAD4 inhibitor, blocked profilin deimination. Neutrophil migration toward leukotriene B4 and toward synovial fluid from a rheumatoid arthritis patient were inhibited by chloramidine, thus supporting the contribution of deimination to chemotaxis. The data, based on a simplified system for integrin activation, imply a mechanism whereby integrin attachment coordinates neutrophil responses to inflammation and orchestrates deimination of nuclear and cytoskeletal proteins. This article is part of the Theo Murphy meeting issue 'The virtues and vices of protein citrullination'. [ABSTRACT FROM AUTHOR]

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]

Epilepsy is one of the most common neurological diseases and affects 1–2% of the world's population. Epilepsy is associated with recurrent and unpredictable seizures. Since 20% of epileptic patients suffer from drug resistance, much research is being done to find new ways to treat epilepsy. One of the new therapies is low-frequency deep brain stimulation (L-DBS). Although this method has an anticonvulsant effect and is used as a new method for the treatment of drug-resistant epilepsy, the mechanism of its action is still unclear. Understanding the changes in the amount of expression of proteins and enzymes in the nervous system following the induction of L-DBS is helpful for understanding the mechanism of effects of L-DBS. In this study, rats were first kindled (Common laboratory model of seizures and epilepsy) by electrical kindling (kindled group) and then received the L-DBS (kindled + L-DBS group). Comparison and analysis of the proteome of kindled and L-DBS-treated rats were performed by comparative proteomics using two-dimensional gel electrophoresis. We observed that electrical kindling induced changes in a series of proteins, that normalized by L-DBS. We identified 23 of these proteins as a sample by MALDI TOF/TOF. The protein–protein interaction network was also drawn for these 23 proteins. The most important pathway attributed to these proteins is the cytoskeleton pathway. Our results showed that the cytoskeleton after destruction by electrical kindling was restored to the normal by L-DBS. [ABSTRACT FROM AUTHOR]

Ammopiptanthus mongolicus, a traditional ethnic medicinal herb, is a rare broad-leaved evergreen shrub in the arid region of central Asia that can survive under extremely low temperatures during winter. In this study, we investigated the cold stress response of A. mongolicus leaves using physiological and proteomic approaches. Cold stress treatment increased the relative electrolyte leakage, proline, and soluble sugar levels and decreased the chlorophyll content in A. mongolicus leaves. Moreover, 93 differentially abundant proteins (DAPs) were identified using two-dimensional electrophoresis, of which 79 were further confirmed via tandem mass spectrometric analysis. The predicted functions of DAPs were mainly associated with photosynthesis in chloroplasts, reactive oxygen species scavenging, defense, and protein synthesis, folding, and degradation. A. mongolicus chitinase A (AmCHIA) is a cold-induced apoplast protein whose transcription is upregulated under cold, osmotic, high-salinity, and mechanical stresses. Recombinant AmCHIA expressed in Escherichia coli exhibits chitinase activity. Here, AmCHIA expression enhanced the cold tolerance of E. coli cells, suggesting that it may contribute to the cold adaptation of A. mongolicus after cold treatment. The present study not only provides important data for understanding the cold stress responses in plants but also serves as a basis for further exploration of the biological functions of cold-induced proteins in A. mongolicus using genetic approaches. [ABSTRACT FROM AUTHOR]

GM risk assessments are crucial for determination and prevention of potential adverse effects through early detection and proper evaluation of intended and potential unintended changes in molecular breeding. In this context, the concept of 'substantial equivalence' has been suggested for the safety tests of GM products for many years. This study evaluated differences between four types of Glycine max crops; transgenic soybean, its non-transgenic counterpart, gamma induced soybean mutant and its parental line, at proteomic level in context of substantial equivalence. The results revealed the ratio of differentially expressed protein spots to total protein spots in mutants compared to their parental line was 50.7%. This ratio was 41.2% in transgenic plants compared to their counterparts. Scatter plot analysis presented those mutant plants showed a wider spread than transgenic plants in terms of distribution of proteins. It was determined that up-regulated proteins in mutant plants have various biological roles in processes such as development, stress response, photorespiration and ribosomal subunit assembly. In transgenic plants, upregulated proteins were shown to have diverse biological roles in processes such as ribosomal subunit assembly, electron transport, amino acid and nucleic acid metabolism and cell wall biogenesis. Although debate on the potential unintended effects focuses on GM organisms, our results point out that the plants that gained new characteristics by various breeding methods should be characterized in all aspects case-by-case. [ABSTRACT FROM AUTHOR]

The objectives of the study are to identify bioactive peptides from goat colostrum and use in-silico analysis to describe their physicochemical properties. Total protein fractions in colostrum (1–3 days), transitional (4–20 days), and mature (after 25 days) goat milk samples were quantified in the biochemical studies. Colostrum (27.26 mg/ml), transitional (12.01 mg/ml), and mature (6.56 mg/ml) protein concentrations were measured. In comparison to the transitional and mature stages of goat milk, the results showed that protein fractions in the colostrum stage have the highest protein concentration. The fat, protein, lactose, total solids, and immunoglobulin percentages in goat colostrum are also high. In 2D electrophoresis, a total of 81 protein spots were observed between 6 and 72 kDa marker regions. Out of these 81 protein spots, about nine prominent quantitative protein spots numbered 12, 23, 25, 30, 34, 60, 75, 77 and 79 were analyzed through MALDI-TOF/MS and they were identified as K casein, Alpha Lactalbumin, Ferritin, Calmodulin dependent protein kinase, Prolactin, Lactoferrin, Lactoperoxidase, Lysozyme and Alpha-S2-casein proteins respectively. The molecular weights of K casein, Lactoferrin, Lactoperoxidase, and Alpha-S2-casein proteins were determined using in silico physicochemical characterisation to be 69564.89 kDa, 21455.34 kDa, 80365.85 kDa, and 26388.09 kDa, respectively. These proteins contain a higher concentration of alpha helices, extended strands, beta twists, and random coils, according to their secondary structure. The findings of this study will aid future research into the isolation of bioactive peptides and their biological roles. [ABSTRACT FROM AUTHOR]

To explore the effect of a chitosan-based preservative solution (10 mg/mL chitosan + 5 mg/mL tea polyphenol + 2 000 U/mL lysozyme) on the change in the muscle proteome of grass carp during cold storage, differential proteomic analysis of fresh fish and nine-day-stored fish with or without (control) preservative treatment was conducted by two-dimensional electrophoresis-mass spectrometry (2DE-MS). The results showed that the 2DE map of the preservative-treated group was not obviously different from that of the fresh group, except for a decrease in the abundance of fast troponin T and an increase in the abundance of slow troponin T and adenylate kinase isoenzyme. However, the 2DE profile of the control group differed greatly from that of the fresh group in both the number and signal intensity of protein spots. Further analysis showed that the abundance of fast troponin T, tropomodulin 4, synaptopodin 2 and pyruvate kinase type M were down-regulated, while the abundance of myosin heavy chain, myosin light chain 1, α-actin, α-tropomyosin and intermediate light meromyosin were up-regulated. Moreover, compared with the preservative-treated group, the abundance of three troponins T were decreased, while the abundance of myosin heavy chain, intermediate light meromyosin, α-actin, α-tropomyosin and creatine kinase M-type were increased in the control group, suggesting that creatine kinase could serve as an indicator for evaluating grass carp muscle spoilage. This study demonstrates that the chitosan-based composite preservative can maintain the stability of grass carp muscle protein during cold storage.

Protein adsorption is essential for determining material biocompatibility and promoting adherent cell growth. In this study, we focused on the a-plane structure of hydroxyapatite (HAp). This a-plane structure closely resembles the crystal plane where apatite is exposed in long bones. We conducted protein adsorption experiments using HAp ceramics with a preferred orientation to a-planes (aHAp), employing bovine serum albumin (BSA), lysozyme, and fetal bovine serum (FBS) as protein models to mimic the in vivo environment. Higher zeta potential and contact angle values were found in aHAp than in HAp ceramics fabricated from commercial HAp powder (iHAp). Bradford-quantified protein adsorption revealed BSA adsorption of 212 ng·mm−2 in aHAp and 28.4 ng mm−2 in iHAp. Furthermore, the Bradford-quantified protein adsorption values for FBS were 2.07 μg mm−2 in aHAp and 1.28 µg mm−2 in iHAp. Two-dimensional electrophoresis (2D-PAGE) showed a higher number of protein-derived major spots in aHAp (37 spots) than in iHAp (12 spots). Mass spectrometry analysis of the resulting 2D-PAGE gels revealed proteins adsorbed on aHAp, including secreted frizzled-related protein 3 and vitamin K epoxide reductase complex 1, which are involved in cellular bone differentiation. Overall, these proteins are expected to promote bone differentiation, representing a characteristic property of aHAp. [ABSTRACT FROM AUTHOR]

The study of an organism's genome, often known as "genomics," has advanced quickly, producing a wealth of publicly accessible genetic data. Despite how valuable the genome is; proteins essentially control most aspects of cell function. Proteomics, or the comprehensive study of proteins, has emerged as an important technology for disease characterization, diagnosis, prognosis, drug development, and therapy. Proteomics technologies are now used to support the diagnosis and treatment of both infectious and noninfectious diseases. Nevertheless, it is more difficult to describe a proteomic profile since a single gene product may result in a number of unique proteins, and proteins have a wider range of chemical configurations. The proteome profiles of a particular organism, tissue, or cell are impacted by a variety of environmental factors, including those triggered by infectious agents. This review intends to highlight the applications of proteomics in the study of disease diagnosis and treatment. In this review, the different technologies used in proteomics studies, like two-dimensional gel electrophoresis, mass spectrometry, and protein microarray as well as biomarker discovery and drug target identification using proteomics, have also been focused on. [ABSTRACT FROM AUTHOR]

In allergic individuals, ingestion of wheat can lead to wheat-dependent exercise-induced anaphylaxis (WDEIA). Many studies have been conducted to find WDEIA allergen–deficient wheat, including by generating omega-5 gliadin antibodies. However, the reported antibodies have not been specific enough to detect omega-5 gliadins encoded on the 1B chromosome. In this study, we generated monoclonal antibodies against the major allergens causing WDEIA, omega-5 gliadins. Using these antibodies (mono-O5B-1C10), we assessed accumulation of omega-5 gliadins in wild-type and nullisomic-tetrasomic (NT) lines of the wheat (Triticum aestivum) varieties Chinese Spring (CS) by one- and two-dimensional gel electrophoresis, followed by Coomassie blue staining or immunoblotting with mono-O5B-1C10. We also tested mono-O5B-1C10 for major omega-5 gliadins in various wheat germplasms. Our results thus demonstrate the specificity of mono-O5B-1C10 for major omega-5 gliadins and potentially useful for identifying of omega-5 gliadin–deficient wheat varieties that should not cause WDEIA. [ABSTRACT FROM AUTHOR]

The number of people suffering from food allergies and intolerances has been increasing in recent years and cereal proteins are the most common food allergens. Pseudocereals represent perspective alternative in nutrition with a positive effect on the human body. The aim of the work was to analyze the proteome of selected varieties of wheat (Triticum aestivum L.), oats (Avena sativa) and buckwheat (Fagopyrum esculentum Moench.) using two-dimensional electrophoresis (2DE) and mass spectrometry in order to detect the presence of potentially allergenic proteins. Using the PDQuest program, 221 protein spots ranging from 4.13 to 9.89 µl with experimental molecular weights from 12.42 kDa to 140 kDa were quantified in 2DE gels of wheat. In the oat sample, 168 protein spots were quantified in the range pI of 4.02 to 9.93 and an experimental molecular weight of 14.81 kDa to 67.96 kDa. Buckwheat proteins were separated on a 2DE gel into 208 protein spots in the 3 to 9.83 pI region with an experimental molecular weight of 10.10 kDa to 115 kDa. By comparing the data with the Allergome database, allergens Tri and 26, Tri and 33, Tri and 36, Tri and alpha Gliadin, Tri and 20 were detected in wheat, Ave s 11S allergens in oats and Fag e 1 allergen in buckwheat. 2DE together with mass spectrometry have been shown to be suitable and sensitive methods for the detection of allergens in food crops. [ABSTRACT FROM AUTHOR]

Background: Non-muscle-invasive bladder cancer (NMIBC) has a high recurrence rate and muscle-invasive bladder cancer (MIBC) has unfavorable outcomes in urothelial bladder cancer (UBC) patients. Complex UBC-related protein biomarkers for outcome prediction may provide a more efficient management approach with an improved clinical outcome. The aim of this study is to recognize tumor-associated proteins, which are differentially expressed in different stages of UBC patients compared non-cancerous tissues. Methods: The proteome of tissue samples of 42 UBC patients (NMIBC n = 25 and MIBC n = 17) was subjected to two-dimensional electrophoresis (2-DE) combined with Liquid chromatography–mass spectrometry (LC–MS) system to identify differentially expressed proteins. The intensity of protein spots was quantified and compared with Prodigy SameSpots software. Functional, pathway, and interaction analyses of identified proteins were performed using geneontology (GO), PANTHER, Reactome, Gene MANIA, and STRING databases. Results: Twelve proteins identified by LC-MS showed differential expression (over 1.5-fold, p < 0.05) by LC-MS, including 9 up-regulated in NMIBC and 3 up-regulated in MIBC patients. Proteins involved in the detoxification of reactive oxygen species and cellular responses to oxidative stress showed the most significant changes in UBC patients. Additionally, the most potential functions related to these detected proteins were associated with peroxidase, oxidoreductase, and antioxidant activity. Conclusion: We identified several alterations in protein expression involved in canonical pathways which were correlated with the clinical outcomes suggested might be useful as promising biomarkers for early detection, monitoring, and prognosis of UBC. [ABSTRACT FROM AUTHOR]

Proteomics tools in the field of equine meat research have been very recently applied to explore the changes in the post-mortem muscle proteome and to discover biomarkers to monitor the variations in its different meat quality traits. The current advances achieved by proteomics in equine meat research are reviewed. Different proteomics techniques (sodium dodecyl sulphate-polyacrylamide gel electrophoresis; two-dimensional polyacrylamide gel electrophoresis; fluorescent two-dimensional difference gel electrophoresis; targeted proteomics; tandem-mass tag labeled proteomics; data-independent analysis proteomics) have been applied in the study of the equine muscle/meat. The studies revealed the biochemical pathways involved in the development of several donkey and horse (foal) meat quality variation. The current knowledge would be useful to develop high-quality products. [ABSTRACT FROM AUTHOR]

Abstract Background Tear proteomic analysis has become an important tool in medical and veterinary research. The tear collection method could influence the tear protein profile. This study aims to evaluate the protein profiles of dog tears collected using microcapillary tubes (MT), Schirmer tear strips (ST), and ophthalmic sponges (OS). Methods The tear samples were collected using MT, ST, and OS. Tear protein profiles were analyzed using two-dimensional electrophoresis (2-DE) and the different protein spots’ expression was compared. Fourteen protein spots were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results Tear protein concentrations ranged from 2.80 to 4.03 μg/μL, with no statistically significant differences among collection methods. Protein expression in each collection method differed in terms of both the number and intensity of the spots. There were 249, 327, and 330 protein spots found from tears collected with MT, ST, and OS, respectively. The proteins albumin, haptoglobin, and lactoferrin identified from OS were found to have higher spot intensities than other methods of collection. The use of MT demonstrated the downregulation of nine proteins. Conclusions The recent study supported that tear protein analysis is affected by different tear collection methods. Although ST is commonly used for tear collection, it provides insufficient information to study particular tear proteins.

Moniliasis caused by the fungus Moniliophthora roreri is one of the most destructive cacao diseases in tropical America. In 2021, the presence of the pathogen was confirmed in cacao plantations in Brazil. This is the first proteomic study of the M. roreri protein profile of ungerminated spores and mycelium compared by 2D SDS‐PAGE (Two‐dimensional gel electrophoresis) associated with mass spectrometry. A total of 446 spots were detected on ungerminated spores gels and 402 spots on M. roreri mycelial gels. A total of 29 proteins were identified from the ungerminated spores, 21 of them were exclusive, and 53 were identified in the mycelium, 16 of them were exclusive. Most of the identified proteins at both development stages were categorized as being involved in metabolic processes, reduction/oxidation processes, and protein synthesis and folding. The interaction networks observed expand the known interactions of these proteins, resulting in eight functional clusters for proteins identified in ungerminated spores and seven clusters for mycelial proteins. In addition, a biological model of ungerminated spores and mycelium with cell location of proteins was constructed. The results contribute to a better understanding of the metabolic mechanisms of M. roreri during dormancy and vegetative development, which can support future studies for strategies to control moniliasis. [ABSTRACT FROM AUTHOR]

The number and identity of proteins and proteoforms presented in a single human cell (a cellular proteome) are fundamental biological questions. The answers can be found with sophisticated and sensitive proteomics methods, including advanced mass spectrometry (MS) coupled with separation by gel electrophoresis and chromatography. So far, bioinformatics and experimental approaches have been applied to quantitate the complexity of the human proteome. This review analyzed the quantitative information obtained from several large-scale panoramic experiments in which high-resolution mass spectrometry-based proteomics in combination with liquid chromatography or two-dimensional gel electrophoresis (2DE) were used to evaluate the cellular proteome. It is important that even though all these experiments were performed in different labs using different equipment and calculation algorithms, the main conclusion about the distribution of proteome components (proteins or proteoforms) was basically the same for all human tissues or cells. It follows Zipf's law and has a formula N = A/x, where N is the number of proteoforms, A is a coefficient, and x is the limit of proteoform detection in terms of abundance. [ABSTRACT FROM AUTHOR]

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