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Additional file 2. Figure S2. Positive and negative control of 1D S-glutathionylation analysis. 1D gel analysis of relative fluorescence intensity corresponding to the levels of protein S-glutathionylation of bull sperm with negative and positive controls. Lane 1—blocking control, sample without substrate specific reduction with GRX1. Lane 2—experimental sample, with GRX1 reduction step. Lane 3—positive control, sample treated with GSSG, an oxidized glutathione donor.

Additional file 4. Figure S3. Representative flow cytometry graphs showing mean fluorescence of Fluo 3-AM corresponding to the level of intracellular calcium in non-capacitated sperm (Non-Cap) (A).

Additional file 1. Figure S1. Positive and negative control of 1D S-nitrosylation analysis. 1D gel analysis of relative fluorescence intensity corresponding to the levels of protein S-nitrosylation of bull sperm with negative and positive controls. Lane 1—blocking control, sample without substrate specific reduction with ascorbate. Lane 2—experimental sample, with ascorbate reduction step. Lane 3—positive control, sample treated with NONOate, a nitric oxide donor.

Additional file 9. Figure S8. Representative histograms showing sperm flagella fluorescence intensity corresponding to the level of tyrosine phosphorylation in non-capacitated sperm (Non-Cap) (A), capacitated sperm (Cap PRDX+) (B) and capacitated sperm with PRDX inhibition (Cap PRDX-) (C).

Additional file 6. Figure S5. Representative flow cytometry graphs showing nitric oxide-positive and negative sperm populations in non-capacitated sperm (Non-Cap) (A), capacitated sperm (Cap PRDX+) (B) and capacitated sperm with PRDX inhibition (Cap PRDX−) (C).

The specific chemical reactivity of thiol groups makes protein cysteines susceptible to reactions with reactive oxygen species (ROS) and reactive nitrogen species (RNS) resulting in the formation of various reversible and irreversible oxidative post-translational modifications (oxPTMs). This review highlights a number of gel-based redox proteomic approaches to detect protein oxPTMs, with particular emphasis on S-nitrosylation, which we believe are currently one of the most accurate way to analyze changes in the redox status of proteins. The information collected in this review relates to the recent progress regarding methods for the enrichment and identification of redox-modified proteins, with an emphasis on fluorescent gel proteomics. Gel-based fluorescent proteomic strategies are low-cost and easy-to-use tools for investigating the thiol proteome and can provide substantial information on redox signaling.

Renewable energy sources and their part in the global energy mix are beneficial to energy diversification and environment protection. However, raw biomass is characterized by low heating value, hydrophilic properties, various mechanical durability, and the logistic challenges related to transportation and storage. One frequently used process of combined biomass valorization is torrefaction and pelletization, which increase the heating value, homogeneity, and hydrophobicity of the fuel. However, industrial clients need fuel characterized by favorable grindability, whereas, the individual clients (householders) need fuel with high mechanical durability. Due to the different expectations of final customers regarding biomass fuel properties, it is necessary to investigate the influence of the torrefaction on the mechanical durability of the pellets. In this paper, five various types of pellets and their torreficates (obtained at a temperature of 200 and 300 °C) were examined. Then the mechanical durability index DU and the grindability of the untreated and torrefied pellets were determined. The results indicated that the mechanical durability of untorrefied pellets is significantly greater than torrefied pellets. Interestingly, no significant differences in mechanical durability between torrefied pellets at 200 and 300 °C were observed, For sunflower husk pellets, the DU index amounted to 95.28 ± 0.72 (untorrefied), 47.22% ± 0.28% (torrefied at 200 °C), and 46.34% ± 0.72% (torrefied at 300 °C). Considering the grindability, as the treatment temperature increased the energy demand for grindability decreased. For example, the grindability of pine tree pellets was 15.96 ± 3.07 Wh·kg−1 (untreated), 1.86 ± 0.31 Wh·kg−1 (torrefied at 200 °C), and 0.99 ± 0.17 Wh·kg−1 (torrefied at 300 °C). The highest difference between raw and torrefied pellets was determined for beetroot pomace pellet: 36.31 ± 2.06 Wh·kg−1 (untreated), 3.85 ± 0.47 Wh·kg−1 (torrefied at 200 °C), and 1.03 ± 0.12 Wh·kg−1 (torrefied at 300 °C).

The aim of our study was to compare quality of fresh and cryopreserved semen of sex-reversed female rainbow trout and sex-reversed female brook trout. The effect of different final sperm concentrations in straws with a constant extender concentration and final glucose concentration on the sperm motility parameters of cryopreserved semen was tested. Furthermore, we examined the effect of post-thaw storage time on sperm motility parameters at optimal sperm and glucose concentrations. The effects of semen cryopreservation from sex-reversed female rainbow trout and sex-reversed female brook trout on sperm fertilizing ability at the sperm-to-egg ratios 500,000:1 and 1,000,000:1 after 0 and 60 min of post-thaw storage of semen were investigated. We have demonstrated that final sperm concentrations in straws as well as final glucose concentrations in extended semen influenced post-thaw sperm motility parameters in both species. The high post-thaw sperm motility was recorded for sperm concentrations within the range of 1.0–4.0 × 109 spermatozoa ml−1 for both species. Furthermore, the glucose concentration appeared to be very important for cryopreservation success and its effect was species-specific. The final glucose concentrations of 0.15 M and 0.19 M, for sex-reversed female rainbow trout and sex-reversed female brook trout, respectively, produced the highest results for sperm motility after cryopreservation. The post-thaw sperm motility was unaffected by 60 and 360 min after thawing for sex-reversed female rainbow trout and sex-reversed female brook trout, respectively. The fertilization rates of cryopreserved semen of sex-reversed female rainbow trout were high and did not differ between the investigated sperm:egg ratios after 0 and 60 min of post-thaw storage. However, fertilization rates of cryopreserved semen of sex-reversed females brook trout were lower and decreased after 60 min of post-thaw storage of semen at sperm: egg ratio 1,000,000:1. In conclusion, our results demonstrated differences in freezability between the semen of sex-reversed female rainbow trout and sex-reversed female brook trout. The semen of both species can be cryopreserved at the final sperm concentration of 3.0 × 109 spermatozoa ml−1. However, the influence of final glucose concentration on cryopreservation success, as well as the duration of post-thaw storage time and fertilization rates, are species-specific. In our opinion, standardizing the procedure of semen cryopreservation presented in this study is a prerequisite for the development of repeatable procedures and the future implementation of cryopreserved semen from sex-reversed females into hatchery practice.

It is widely recognized that quality of spermatozoa in sexed semen (SS) samples is not as great as for conventional, non-sexed semen (NS). There are differences in qualitative and biochemical variables between spermatozoa in NS and SS. Information, however, is lacking on molecular differences, especially concerning spermatozoa proteomic differences is SS and NS. The objective of this study was to compare commercially available NS and SS bull semen by evaluating sperm quality variables in conjunction with a proteomics approach. Results from flow cytometry and computer-assisted sperm analyses indicated there was less sperm motility, viability, mitochondrial potential and acrosome integrity in sperm from SS. Results from proteomic analysis indicated sperm from NS and SS samples were characterized by different protein profiles. There was identification of 70 sperm proteins that differed in abundance and six protein spots that were different in extent of carbonylation. Sperm from SS had altered structures of enzymes involved in glycolysis, oxidative phosphorylation and maintenance of a constant adenylate energy charge. Furthermore, sperm from SS had alterations of several flagella substructures, especially outer dense fiber proteins, which were less abundant and more carbonylated than in sperm from NS. In sperm of SS, compared with NS, there were differences in abundance of proteins involved in capacitation, acrosome reaction and sperm-egg fusion as well as lesser abundances of sperm surface proteins. Results enable a greater understanding of differences between sperm from NS and SS samples, thereby contributing to development of improved protocols for more effective protection of sexed spermatozoa during processing.

Artificial insemination with cryopreserved semen enables affordable, large-scale dissemination of gametes with superior genetics. However, cryopreservation can cause functional and structural damage to spermatozoa that is associated with reactive oxygen species (ROS) production, impairment of sperm motility and decreased fertilizing potential, but little attention has been paid to protein changes. The goal of this study was to investigate the oxidative modifications (measured as carbonylation level changes) of bull spermatozoa proteins triggered by the cryopreservation process. Flow cytometry and computer-assisted sperm analysis were used to evaluate changes in viability, ROS level and motility of spermatozoa. Western blotting, in conjunction with two-dimensional electrophoresis (2D-oxyblot) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight spectrometry, was employed to identify and quantify the specifically carbonylated spermatozoa proteins. Cryopreservation decreased motility and viability but increased the number of ROS-positive cells. We identified 11 proteins (ropporin-1, outer dense fiber protein 2, glutathione S-transferase, triosephosphate isomerase, capping protein beta 3 isoform, actin-related protein M1, actin-related protein T2, NADH dehydrogenase, isocitrate dehydrogenase, cilia- and flagella-associated protein 161, phosphatidylethanolamine-binding protein 4) showing differences in protein carbonylation in response to cryopreservation. The identified proteins are associated with cytoskeleton and flagella organization, detoxification and energy metabolism. Moreover, almost all of the identified carbonylated proteins are involved in capacitation. Our results indicate for the first time that cryopreservation induces oxidation of selected sperm proteins via carbonylation. We suggest that carbonylation of sperm proteins could be a direct result of oxidative stress and potentially lead to disturbances of capacitation-involved proteins or could indicate cryopreservation-induced premature capacitation.

The existing knowledge on the bull seminal vesicle proteome, a major seminal plasma constituent, and its relationship with seminal plasma is limited. This knowledge is prerequisite for a better understanding of seminal plasma variability, which is linked to semen quality. The objective of this study was to characterize the proteomes of seminal vesicle fluid and seminal plasma and to compare them to better understand the origin of seminal plasma proteins. We collected ejaculates and seminal vesicle fluid postmortem from 6 mature Holstein Friesian bulls. We performed the analysis and identification of proteins using 2-dimensional electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. We identified 105 proteins in bull seminal vesicle fluid and 88 proteins in seminal plasma. For both seminal vesicles and seminal plasma proteins described in our study, top biological functions were cellular movement, cell death and survival, and cellular growth and proliferation. Additionally, seminal vesicle fluid proteins were involved in protein degradation and synthesis. Seminal plasma proteins were also involved in cellular assembly and organization and cell-to-cell signaling and interactions. Proteins of both fluids were involved in the following canonical pathways: glycolysis, gluconeogenesis, liver X receptor/farnesoid X receptor, and farnesoid X receptor/retinoid X receptor activation. Additionally, seminal vesicle fluid proteins appeared to be involved in oxidative stress response mediated by nuclear factor E2-related factor 2. Our results described the bull seminal vesicle fluid proteome for the first time and allowed for significant expansion of the current knowledge on the bull seminal plasma proteome. Moreover, analysis indicated that both bull seminal vesicle fluid and seminal plasma proteomes contained interconnected protein groups related to protective functions, glycolysis, and the morphology and physiology of the spermatozoa. These proteins and their interactions could be targeted in future research.

W artykule przedstawiono budowę i parametry techniczne sekcji obudowy zmechanizowanej HYDROMEL-16/35-POz. Omówiono szczegóły i powody przeprowadzonych modernizacji. Przedstawiono najważniejsze informacje dotyczące warunków geologiczno-górniczych w kolejnych lokalizacjach sekcji obudowy.

Additional file 2: Figure S1. Search Tool for the retrieval of Interacting Genes/Proteins (STRING) analysis of identified proteins (A) at 24 h and (B) 48 h compared to 8 h of the cultivation. Full proteins name can be found in the UniProtKB database.

Nanoporous anodic aluminum oxide was formed in sulfuric acid with addition of indigo carmine. During anodizing, the organic dye was incorporated into the porous oxide walls. X-ray photoelectron spectroscopy revealed the presence of nitrogen and sulfur in the anodic aluminum oxide. Two types of incorporated sulfur were found: belonging to the sulfate anions SO42 − of the electrolyte and belonging to the C-SO3− side groups of the indigo carmine. Raman spectroscopy confirmed the incorporation and showed that the inorganic–organic hybrid material inherited optical properties from the indigo carmine. Typical modes from pyrrolidone rings, unique for indigo carmine in the investigated system (650 and 1585 cm− 1), were found to be the strongest for the greatest anodizing voltages used. Despite the indigo carmine incorporation, the morphology of the oxide is still nanoporous and its geometry is still tuned by the voltage applied during aluminum anodization. This work presents an inexpensive and facile approach to doping an inorganic oxide material with organic compounds.

During semen cryopreservation, spermatozoa are exposed to physical and chemical stressors that result in their functional and structural damage. Growing evidence suggests that most cryoinjuries result from oxidative stress accompanying sperm cryopreservation. Elevated amounts of reactive oxygen species (ROS) generated during cryopreservation can react with sperm macromolecules, including proteins. The goal of this study was to investigate the oxidative modifications (measured as carbonylation level changes) of carp spermatozoa proteins triggered by the cryopreservation process. Flow cytometry and computer-assisted sperm analysis were used to evaluate changes in viability, ROS level, and motility of spermatozoa. The spermatozoa proteins that were specifically carbonylated were identified and quantified by Western blotting, in conjunction with 2-dimensional electrophoresis (2D-oxyblot) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Cryopreservation decreased spermatozoa motility (P < 0.01) and viability (P < 0.0001) and significantly increased (P < 0.0001) the number of ROS-positive cells. We identified 25 protein spots, corresponding to 19 proteins, with increases (P < 0.05) in carbonylation level due to freezing/thawing. The identified proteins are involved in motility, metabolism, calcium-ion binding, signal transduction, protein folding, and intracellular transport. The results suggest that carbonylation of flagellar proteins can result in motility disorders and may contribute to the reduced percentage of motile spermatozoa and disturbances in movement trajectory after sperm cryopreservation. Moreover, cryopreservation may contribute to impaired cellular respiration, ATP regeneration, disturbances of Ca(2+) turnover, unfolding of cytoplasmic or histone proteins, disturbances of cell signaling and intracellular transport, and reduced membrane stability. Our results contribute to the knowledge concerning cryoinjury and to further development of a modified cryopreservation procedure aimed at minimizing oxidative damage of carp sperm proteins.