Collagen is the most abundant protein in the animal and human bodies, and it is not exempt from this aging phenomenon. Some age-related changes may appear on collagen sequences, such as increased surface hydrophobicity, the appearance of post-translational modifications, and amino acids racemization. This study has shown that the protein hydrolysis under deuterium conditions is privileged to limit the natural racemization during the hydrolysis. Indeed, under the deuterium condition, the homochirality of recent collagens is preserved whose amino acids are found in their L-form. However, in aging collagen, a natural amino acid racemization was observed. These results confirmed that the % d-amino acids are progressive according to age. The collagen sequence is degraded over time, and a fifth of the sequence information is lost during aging. Post-translational modifications (PTMs) in aging collagens can be a hypothesis to explain the modification of the hydrophobicity of the protein with the decrease of hydrophilic groups and the increase of hydrophobic groups. Finally, the exact positions of d-amino acids and PTMs have been correlated and elucidated.
- MeSH
- aminokyseliny * chemie MeSH
- chromatografie kapalinová MeSH
- deuterium chemie MeSH
- kolagen MeSH
- lidé MeSH
- posttranslační úpravy proteinů MeSH
- proteomika * MeSH
- stárnutí MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Actin is a fundamental member of an ancient superfamily of structural intracellular proteins and plays a crucial role in cytoskeleton dynamics, ciliogenesis, phagocytosis, and force generation in both prokaryotes and eukaryotes. It is shown that actin has another function in metazoans: patterning biosilica deposition, a role that has spanned over 500 million years. Species of glass sponges (Hexactinellida) and demosponges (Demospongiae), representatives of the first metazoans, with a broad diversity of skeletal structures with hierarchical architecture unchanged since the late Precambrian, are studied. By etching their skeletons, organic templates dominated by individual F-actin filaments, including branched fibers and the longest, thickest actin fiber bundles ever reported, are isolated. It is proposed that these actin-rich filaments are not the primary site of biosilicification, but this highly sophisticated and multi-scale form of biomineralization in metazoans is ptterned.
- MeSH
- aktiny * MeSH
- kostra MeSH
- oxid křemičitý * chemie MeSH
- sklo MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Saliva contains possible biomarkers that are associated with dental caries. The present study aimed to analyse differences in the abundance of proteins in the saliva between caries-positive (CP; N = 15) and caries-free (CF; N = 12) males and to compare differences in the abundance of proteins between two saliva sample fractions (supernatant and pellet). We found 14 differently significantly expressed proteins in the CF group when comparing the supernatant fractions of the CP and CF groups, and three proteins in the pellet fractions had significantly higher expression in the CP group. Our results indicate very specific protein compositions of the saliva in relation to dental caries resistance (the saliva of the CP group mainly contained pellet proteins and the saliva of the CF group mainly contained supernatant proteins). This was the first time that the saliva pellet fraction was analysed in relation to the dental caries status. We detected specific calcium-binding proteins that could have decalcified enamel in the saliva pellet of the CP group. We also observed significantly up-regulated immune proteins in the saliva supernatant of the CF group that could play an important role in the caries prevention. The particular protein compositions of the saliva pellet and supernatant in the groups with different susceptibilities to tooth decay is a promising finding for future research.
- MeSH
- lidé MeSH
- náchylnost k zubnímu kazu * MeSH
- proteomika * MeSH
- slinné proteiny a peptidy analýza imunologie metabolismus MeSH
- sliny chemie imunologie metabolismus MeSH
- zubní kaz imunologie metabolismus prevence a kontrola MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Lithium (Li) is a typical mood stabilizer and the first choice for treatment of bipolar disorder (BD). Despite an extensive clinical use of Li, its mechanisms of action remain widely different and debated. In this work, we studied the time-course of the therapeutic Li effects on ouabain-sensitive Na+/K+-ATPase in forebrain cortex and hippocampus of rats exposed to 3-day sleep deprivation (SD). We also monitored lipid peroxidation as malondialdehyde (MDA) production. In samples of plasma collected from all experimental groups of animals, Li concentrations were followed by ICP-MS. The acute (1 day), short-term (7 days) and chronic (28 days) treatment of rats with Li resulted in large decrease of Na+/K+-ATPase activity in both brain parts. At the same time, SD of control, Li-untreated rats increased Na+/K+-ATPase along with increased production of MDA. The SD-induced increase of Na+/K+-ATPase and MDA was attenuated in Li-treated rats. While SD results in a positive change of Na+/K+-ATPase, the inhibitory effect of Li treatment may be interpreted as a pharmacological mechanism causing a normalization of the stress-induced shift and return the Na+/K+-ATPase back to control level. We conclude that SD alone up-regulates Na+/K+-ATPase together with increased peroxidative damage of lipids. Chronic treatment of rats with Li before SD, protects the brain tissue against this type of damage and decreases Na+/K+-ATPase level back to control level.
- MeSH
- antimanika farmakologie terapeutické užití MeSH
- hipokampus účinky léků metabolismus MeSH
- kompetitivní vazba účinky léků MeSH
- krysa rodu rattus MeSH
- lithiumkarbonát farmakologie MeSH
- malondialdehyd metabolismus MeSH
- ouabain metabolismus MeSH
- peroxidace lipidů účinky léků MeSH
- potkani Wistar MeSH
- přední mozek účinky léků enzymologie metabolismus MeSH
- sodíko-draslíková ATPasa metabolismus MeSH
- spánková deprivace farmakoterapie enzymologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Eggshell colour, quality, and biosafety of table eggs are of significant commercial interest. To date, there have been few studies investigating the relationship between eggshell pigmentation and internal egg quality in commercially bred birds. Moreover, the genetic basis and mechanisms behind the effects of extrinsic factors on deposition of antimicrobial compounds in egg white and eggshell pigments are not fully understood. In the present study, we evaluate the effect of chicken breed identity, eggshell pigmentation and the role of extrinsic factors (year and breeder identity) on variability in the concentrations of 2 major egg white antimicrobial proteins (AMPs), lysozyme (LSM), and ovotransferrin (OVOTR), across 23 traditional chicken breeds. We found that chicken breed identity and eggshell pigmentation explained most variability in the concentration of egg white LSM and OVOTR. Year and breeder identity were also significant predictors of egg white LSM and OVOTR variability, and showed selective effects on the deposition of both AMPs in egg white. We also documented a positive correlation between concentration of egg white LSM and eggshell cuticle protoporphyrin in tinted and dark brown eggs, but not in brown, white, and blue eggs. We assume that a combination of both intrinsic genetic and hormonally regulated extrinsic factors is responsible for this relationship and for the variability in egg white AMPs. In this study, we demonstrate the existence of a relationship between eggshell pigmentation and egg white AMPs content in the eggs of traditional chicken breeds that may advertise the egg's antimicrobial potential and biosafety. These findings provide novel insights into the relationship between eggshell pigmentation and egg internal quality and may stimulate the recovery and exploitation of traditional chicken breeds for egg production, where the demands for egg quality and biosafety, in conjunction with animal welfare, are a priority.
- MeSH
- imunomodulace genetika MeSH
- kationické antimikrobiální peptidy genetika metabolismus MeSH
- kur domácí genetika imunologie metabolismus MeSH
- pigmentace MeSH
- ptačí proteiny genetika metabolismus MeSH
- vaječná skořápka chemie MeSH
- vaječné proteiny genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The dentin-enamel junction (DEJ) is the border where two different mineralized structures - enamel and dentin - meet. The protein-rich DEJ, together with the inner enamel region of mature teeth, is known to exhibit higher fracture toughness and crack growth resistance than bulk phase enamel. However, an explanation for this behavior has been hampered by the lack of compositional information for the DEJ and the adjacent enamel organic matrix (EOM). We studied proteomes of the DEJ and EOM of healthy human molars and compared them with dentin and enamel proteomes from the same teeth. These tissues were cut out of tooth sections by laser capture microdissection, proteins were extracted and cleaved by trypsin, then processed by liquid chromatography coupled to tandem mass spectrometry to analyze the proteome profiles of these tissues. This study identified 46 proteins in DEJ and EOM. The proteins identified have a variety of functions, including calcium ion-binding, formation of extracellular matrix, formation of cytoskeleton, cytoskeletal protein binding, cell adhesion, and transport. Collagens were identified as the most dominant proteins. Tissue-specific proteins, such as ameloblastin and amelogenin, were also detected. Our findings reveal new insight into proteomics of DEJ and EOM, highly mineralized tissues that are obviously difficult to analyze.
- MeSH
- chromatografie kapalinová MeSH
- dentin * MeSH
- lidé MeSH
- mikrodisekce MeSH
- moláry * MeSH
- proteiny metabolismus MeSH
- proteom analýza MeSH
- proteomika metody MeSH
- tandemová hmotnostní spektrometrie MeSH
- zubní sklovina * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The review is focused on the latest developments in the analysis of proteins and peptides by capillary electrophoresis techniques coupled to mass spectrometry. First, the methodology and instrumentation are overviewed. In this section, recent progress in capillary electrophoresis with mass spectrometry interfaces and capillary electrophoresis with matrix-assisted laser desorption/ionization is mentioned, as well as separation tasks. The second part is devoted to applications-mainly bottom-up and top-down proteomics. It is obvious that capillary electrophoresis with mass spectrometry methods are well suited for peptide and protein analysis (proteomic research) and it is described how these techniques are complementary and not competitive with the often used liquid chromatography with mass spectrometry methods.
Idiopathic pes equinovarus (clubfoot) is a congenital deformity of the feet and lower legs. Clubfoot belongs to a group of fibro-proliferative disorders but its origin remains unknown. Our study aimed to achieve the first complex proteomic comparison of clubfoot contracted tissue of the foot (medial side; n = 16), with non-contracted tissue (lateral side; n = 13). We used label-free mass spectrometry quantification and immunohistochemistry. Seven proteins were observed to be significantly upregulated in the medial side (asporin, collagen type III, V, and VI, versican, tenascin-C, and transforming growth factor beta induced protein) and four in the lateral side (collagen types XII and XIV, fibromodulin, and cartilage intermediate layer protein 2) of the clubfoot. Comparison of control samples from cadavers brought only two different protein concentrations (collagen types I and VI). We also revealed pathological calcification and intracellular positivity of transforming growth factor beta only in the contracted tissue of clubfoot. Most of the 11 differently expressed proteins are strongly related to the extracellular matrix architecture and we assume that they may play specific roles in the pathogenesis of this deformity. These proteins seem to be promising targets for future investigations and treatment of this disease. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.
- MeSH
- dítě MeSH
- extracelulární matrix - proteiny metabolismus MeSH
- hmotnostní spektrometrie MeSH
- kalcinóza MeSH
- lidé MeSH
- pes equinovarus etiologie metabolismus MeSH
- předškolní dítě MeSH
- prospektivní studie MeSH
- proteom MeSH
- transformující růstový faktor beta metabolismus MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Saiga horn extracts were analyzed with the goal of obtaining new information about compounds present in it. The purpose of this study is to find synthetic alternatives to Saiga horn extract, which is used in traditional Chinese medicine, by identifying potentially biologically active compounds in the extracts. Using high-performance liquid chromatography coupled with high-resolution mass spectrometry, we have been able to identify a series of short-chain polyhydroxybutyrates in alcoholic extracts of Saiga horn. Optimized high-performance liquid chromatography coupled with tandem mass spectrometry methods for analysis of short-chain poly-3-hydroxybutyrates were developed and subsequently applied to investigate Saiga horn extract for the presence of these compounds, which might explain its biological actions, particularly for its antipyretic and procoagulant properties.
The proteins and pigment of the eggshell of the Siamese crocodile (Crocodylus siamensis) were analysed. For proteomic analysis, various decalcification methods were used when the two main surface layers were analyzed. These layers are important for antimicrobial defense of egg (particularly the cuticle). We found 58 proteins in both layers, of which 4 were specific for the cuticle and 26 for the palisade (honeycomb) layer. Substantial differences between proteins in the eggshell of crocodile and previously described birds' eggshells exist (both in terms of quality and quantity), however, the entire proteome of Crocodilians has not been described yet. The most abundant protein was thyroglobulin. The role of determined proteins in the eggshell of the Siamese crocodile is discussed. For the first time, the presence of porphyrin pigment is reported in a crocodilian eggshell, albeit in a small amount (about 2 to 3 orders of magnitude lower than white avian eggs).
