Proteomic profile of seminal plasma from Pêga donkeys (Equus asinus) with high sperm motility and vigor: Implications for assisted reproduction.

Donkeys of the Pêga breed (Equus asinus) have been used for two centuries to produce breeding stock and create hybrids for labor and transport in southeast Brazil, and for exporting meat and milk to other countries. Furthermore, they are used in competitions, as they are docile and easy to handle. However, assisted reproduction success rates for frozen donkey semen are remarkably low, with no standardized method for cryopreserving sperm after removal of seminal plasma. This work aims to reveal the biological involvement of seminal plasma proteins from Pêga donkeys in aiding the development of assisted reproduction. This study was carried out with 14 ejaculates collected every eight days, throughout the breeding season, from three healthy fertile Pêga donkeys, with an average age of four years. After confirming the high freezability of fresh semen by evaluating quality parameters, the seminal plasma was separated by centrifugation and an aliquot from each collection was microfiltered and frozen. A label-free technique followed by LC-MS/MS analysis applied to pools of seminal plasma samples from each animal revealed 522 proteins in the proteomic profile, of which 49.8 % (260 proteins) are related to cellular energy transformation, and many proteins involved in reproduction (76), spermatogenesis (38), fertilization (29), among other biological process. By comparison with literature, Pêga donkeys share many proteins with donkeys of Dezhou breed that present great potential as fertility biomarkers. Our results showed proteins positively related to fertilization for different breeds of donkeys around the world, helping to enhance the assisted reproduction of Pêga donkeys.

Human milk proteins differentiate over the sex of newborns and across stages of lactation.

BACKGROUND & AIMS: Human milk (HM) is a complete food that meets the nutritional and energy demands of the newborns. It contains numerous bioactive components, including functional proteins. Variations in HM energy and lipid content have already been reported related to the newborn's sex, but differences between protein profiles are still scarce. This work aimed to identify differences between HM proteins produced by mothers of female and male newborns, in the lactation stages of colostrum and mature milk, and the metabolic pathways involved. METHODS: A total of 98 HM samples were collected from 39 lactating women and classified according to the newborn's sex, stages of lactation, and three mothers' age groups, and evaluated about protein concentration and one-dimensional electrophoretic profile. Next, to assess samples with the greatest differences, the HM proteins regarding the newborn's sex and the stages of lactation were compared using nano-LC-MS/MS, in 24 HM samples randomly rearranged into four groups: female and male infants, and colostrum and mature milk. Functional classification, metabolic pathways, and protein interaction networks were analyzed by Gene Ontology, KEGG, and STRING, respectively. RESULTS: The soluble protein content of HM decreased throughout lactation, with differences regarding isolated factors, such as mothers' age group, child's sex and stages of lactation, and also in terms of their interactions. A total of 146 proteins were identified, 42 of which showed different abundances over the sexes of newborns and 53 between the stages of lactation. In general, proteins related to metabolic processes were up-regulated for mothers of male infants and in the mature stage of lactation, while proteins related to defense were up-regulated in mothers of female infants and in the colostrum phase. CONCLUSION: This study indicated that there are differentiated and specific nutritional and defense needs of newborns, by sex and by lactation phase, which is highly relevant for a more appropriate supply of food to infants receiving HM from donor mothers.

Dataset of seminal plasma proteome of Nellore bulls with high and low percentage of abnormalities sperm.

OBJECTIVES: Bovine seminal plasma proteins perform several functions related to sperm function. Changes in the expression pattern or abundance of seminal proteins are related to changes in the fertilizing capacity of bulls. Considering the role of seminal plasma proteins in sperm function and animal reproduction, we investigated changes in the protein abundance profile in response to sperm morphological changes using a proteomic approach. DATADESCRIPTION: In our present investigation, we employed liquid chromatography coupled with mass spectrometry to elucidate the proteomic composition of seminal plasma obtained from Nellore bulls exhibiting varying percentages of sperm abnormalities. Following semen collection, seminal plasma was promptly isolated from sperm, and proteins were subsequently precipitated, enzymatically digested using porcine trypsin, and subjected to analysis utilizing the Acquity nano UHPLC System in conjunction with a mass spectrometer. This dataset encompasses a total of 297 proteins, marking the inaugural instance in which a comparative profile of seminal plasma proteins in young Nellore bulls, categorized by their sperm abnormality percentages, has been delineated using LC-MS/MS. The comprehensive nature of this dataset contributes pivotal proteomic insights, representing a noteworthy advancement in our understanding of the reproductive biology of the Nellore breed.

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense.

Background: Plant protease inhibitors play a crucial role in inhibiting proteases produced by phytopathogens and exhibiting inhibitory effects on nematodes, fungi, and insects, making them promising candidates for crop protection. Specifically, carboxypeptidase inhibitors, a subset of proteinase inhibitors, have been extensively studied in potato and tomato of Solanaceae plant family. However, further research is needed to fully understand the functions and biotechnological potential of those inhibitors in plants. This work aimed to in silico characterize carboxypeptidase inhibitors from Solanaceae as potential antimicrobial and defense agents focused on biotechnological targets. Methods: The methodology employed involved search in UniProt, PDB, KNOTTIN, NCBI, and MEROPS databases for solanaceous carboxypeptidase inhibitors, phylogenetic relationships and conservation patterns analyzes using MEGA-X software and Clustal Omega/MView tools, physicochemical properties and antimicrobial potential prediction using ProtParam, ToxinPred, iAMPred, and APD3 tools, and structural features prediction using PSIPRED. Results and discussion: A systematic literature search was conducted to identify relevant studies on Solanaceae carboxypeptidase inhibitors and their activities against pathogens. The selected studies were reviewed and the main findings compiled. The characterization of Solanaceae carboxypeptidase inhibitors proposed for the first time the global sequence consensus motif CXXXCXXXXDCXXXXXCXXC, shedding light on carboxypeptidase inhibitors distribution, sequence variability, and conservation patterns. Phylogenetic analysis showed evolutionary relationships within the Solanaceae family, particularly in Capsicum, Nicotiana, and Solanum genera. Physicochemical characteristics of those peptides indicated their similarity to antimicrobial peptides. Predicted secondary structures exhibited variations, suggesting a broad spectrum of action, and studies had been demonstrated their activities against various pathogens. Conclusion: Carboxypeptidase inhibitors are being proposed here as a new subclass of PR-6 pathogenesis-related proteins, which will aid in a focused understanding of their functional roles in plant defense mechanisms. These findings confirm the Solanaceae carboxypeptidase inhibitors potential as defense agents and highlight opportunities for their biotechnological applications in pathogen control.

Relative Abundance of Spermadhesin-1 in the Seminal Plasma of Young Nellore Bulls Is in Agreement with Reproductive Parameters.

This study aimed to evaluate the proteomic profile of seminal plasma from young Nellore bulls. We used 20 bulls aged between 19.8 and 22.7 months, divided into two groups according to the results of the Breeding Soundness Evaluation (BSE): approved (FIT n = 10) and not approved (UNFIT n = 10). The scrotal perimeter was measured and a semen collection was performed through electroejaculation. The percentage of sperm motility, mass motility, and sperm vigor were calculated using conventional microscopy, and the percentage of sperm abnormalities was calculated using phase-contrast microscopy of all ejaculates. Seminal plasma was separated from spermatozoa using centrifugation and processed for proteomic analysis by LC-MS/MS. Seminal plasma proteins were identified using MASCOT Daemon software v.2.4.0 and label-free quantification analysis was carried out by SCAFFOLD Q+ software v.4.0 using the Exponentially Modified Protein Abundance Index (emPAI) method. Functional classification of proteins was performed based on their genetic ontology terms using KOG. Functional cluster analysis was performed on DAVID. There were no differences in scrotal perimeter and physical semen characteristics between FIT and UNFIT groups of bulls. The percentage of sperm abnormalities was higher (p < 0.05) in the UNFIT group of bulls. A total of 297 proteins were identified for the two groups. There were a total of 11 differentially abundant proteins (p < 0.05), two of them more abundant in FIT bulls (Spermadhesin-1 and Ig gamma-1 chain C region) and nine in UNFIT bulls (Vasoactive intestinal peptide, Metalloproteinase inhibitor 2, Ig lambda-1 chain C regions, Protein FAM3C, Hemoglobin beta, Seminal ribonuclease, Spermadhesin 2, Seminal plasma protein BSP-30kDa, and Spermadhesin Z13). Spermadhesin-1 was the protein with the highest relative abundance (36.7%) in the seminal plasma among all bulls, corresponding to 47.7% for the FIT bulls and 25,7% for the UNFIT bulls. Posttranslational modification, protein turnover, and chaperones were the functional categories with the highest number of classified proteins. Protein functional annotation clusters were related to Phospholipid efflux, ATP binding, and chaperonin-containing T-complex. The differentially abundant proteins in the group of FIT bulls were related to sperm capacitation and protection against reactive species of oxygen. In contrast, differentially expressed proteins in the group of UNFIT bulls were related to motility inhibition, intramembrane cholesterol removal and oxidative stress. In conclusion, the proteomic profile of the seminal plasma of FIT bulls presents proteins with participation in several biological processes favorable to fertilization, while the proteins of the seminal plasma of UNFIT bulls indicate a series of alterations that can compromise the fertilizing capacity of the spermatozoa. In addition, the relative abundance of spermadhesin-1 found in the seminal plasma of young Nellore bulls could be studied as a reproductive parameter for selection.

Oxidative stress associated with proteomic and fatty acid profiles of sperm from Nellore bulls at rest†.

Sexual rest is a transient condition, which compromises conception rates, characterized by large volumes of ejaculate with high percentages of dead sperm observed in bulls. The biochemical mechanisms leading to this ejaculate pattern are not fully understood. Six adult resting Nellore bulls were submitted to Breeding Soundness Evaluation by four consecutive semen collections through the electroejaculation method during a 30 min period. Each ejaculate had its semen phenotypic parameters; morphology and physical aspects were evaluated. To assess enzymatic activity (superoxide dismutase, catalase, and glutathione S-transferase), lipid peroxidation (concentrations of malondialdehyde and nitric oxide), fatty acid, and proteomic profile aliquots of spermatozoa from the first and fourth ejaculates were used. All sperm parameters differed between the first and fourth ejaculates. Spermatozoa from the first ejaculate showed lower enzymatic activity and a higher concentration of lipid peroxidation markers. Among the 19 identified fatty acids, 52.7% are polyunsaturated. Relative abundance analysis showed that C12:0 and C18:0 fatty acids differed between the first and fourth ejaculates, being the fourth ejaculate richer in spermatozoa. The proteomics analysis identified a total of 974 proteins in both sample groups (first and fourth ejaculates). The majority of identified proteins are related to cellular processes and signaling. Quantitative proteomics showed 36 differentially abundant proteins, 6 up-regulated proteins in the first ejaculate, and 30 up-regulated proteins in the fourth ejaculate. Spermatozoa from bulls at sexual rest have less antioxidant capacity, causing changes in their fatty acid composition and protein profile, which generates the observed sperm pattern and lower fertilization capacity.

Proteoforms of ß-Casein, α-s1-Casein, α-Lactalbumin, Serum Albumin, and Lactotransferrin Identified in Different Lactation Phases of Human Milk by Proteomics Approach.

SCOPE: Human milk (HM) has a wide range of proteins with biological and nutritional functions, essential for newborns. The roles of proteins and their proteoforms in HM are not fully understood. This study aims to assess, by 2-DE proteomics, the differential proteoforms in HM, present in colostrum (COL), transition (TRA), and mature milk (MAT), aiming to contribute to understanding neonates' protein needs. METHODS AND RESULTS: HM samples are collected from 39 healthy lactating women. COL presents the higher concentration of essential amino acids. After MALDI-MS/MS and bioinformatics analysis, proteoforms are differentially detected. Abundances of ß-casein (CSN2), α-s1 casein, and α-lactalbumin (LALBA) are higher in MAT; CSN2s are found in 11 spots and the isoforms increase in size as the pI becomes more acidic; regarding LALBA, two variant forms are found with different abundances in TRA and MAT; CSN2, LALBA, lactotransferrin (LTF), and serum albumin forms are present in all lactation phases. CONCLUSION: This study reveals differential proteoforms in COL involved in tissue growth and body development, besides essential amino acids, and, in MAT, involved in muscle mass gain, strengthening of the immune system, and energy production. The results provide new insight about proteoforms involved in maturation of the newborn's organs and systems.

Kallikrein proteoforms and reproductive parameters in stallion are conditioned by climate.

Horses are seasonal polyoestrous animals, and the photoperiod is the main factor modulating their reproductive activity. There is no consensus on the andrological and biochemical factors that influence breeding seasonality. To assess the involvement of climate in reproduction, Mangalarga Marchador stallions were monitored over 1 year regarding semen quality and seminal plasma proteome. Here, we show that kallikrein (KLKs) proteoforms in seminal plasma are involved in climate conditioning of reproduction. During the breeding season, greater abundance and different types of KLKs occurred simultaneously to lower sperm motility, greater semen volumes and higher concentrations of glucose and cholesterol. Considering that vasodilation due to activation of the kallikrein-kinin system and the consequent inhibition of the renin-angiotensin system may be associated with lower sperm motility, unravelling the involvement of KLK proteoforms in reproductive seasonality is a priority in horse breeding.

Protein expression profiling of Staphylococcus aureus in response to the bacteriocin bovicin HC5.

Alternative strategies to antibiotic treatment are required to inhibit pathogens, including Staphylococcus aureus. Bacteriocins, such as the lantibiotic bovicin HC5, have shown potential to control pathogens. This study aims to evaluate the stress response of S. aureus to bovicin HC5 using a proteomic approach. Sublethal concentrations of the bacteriocin repressed the synthesis of 62 cytoplasmic proteins, whereas 42 proteins were induced in S. aureus COL. Specifically, synthesis of several proteins involved in amino acid biosynthesis, mainly products of ilv-leu operon, and DNA metabolism, such as DNA polymerase I, decreased following bovicin treatment while proteins involved in catabolism, mainly tricarboxylic acid cycle metabolism, and chaperones were over-expressed. The levels of CodY and CcpA, important regulators involved in the stationary phase adaptation and catabolite repression, respectively, also increased in the presence of the bacteriocin. These results indicate that stress caused by the sublethal concentration of bovicin HC5 in the cell membrane results in growth reduction, reduced protein synthesis, and, at the same time, enhanced the levels of chaperones and enzymes involved in energy-efficient catabolism in an attempt to restore energy and cell homeostasis. These results bring relevant information to amplify the knowledge concerning the bacterial physiological changes in response to the stress caused by the cell exposition to bovicin HC5. New potential targets for controlling this pathogen can also be determined from the new protein expression pattern presented. KEY POINTS: • Bovicin HC5 changed the synthesis of cytoplasmic proteins of S. aureus. • Bovicin HC5 interfered in the synthesis of proteins of amino acids biosynthesis. • Synthesis of chaperones enhanced in the presence of sublethal dosage of bovicin HC5.

Movimentos de profissionalização histórica: a relação entre as catástrofes sociais e a enfermagem moderna / Historical professionalization movements: the relationship between social disasters and modern nursing / Movimientos de profesionalización histórica: la relación entre las catástrofes sociales y la enfermería moderna

Resumo Contexto: É em contexto de catástrofes sociais que a afirmação socioprofissional do enfermeiro se constitui uma referência. Objetivo: Dimensionar a relação entre as catástrofes sociais e a relevância do trabalho realizado por determinadas enfermeiras, no final do século XIX, com contributo para a construção/evolução da enfermagem enquanto profissão. Metodologia: Embora as opções metodológicas na validação do caráter científico deste estudo histórico assentem na historiografia, os factos narrados e interpretados foram contextualizados em função da época histórica, permitindo o emergir de novos significados que possibilitem definir no próprio tecido documental, unidades, conjuntos, séries, relações inovadoras. Resultados: Foram várias as enfermeiras que contribuíram para a evolução da enfermagem enquanto ciência. Experienciaram a sua profissão em contextos com um denominador comum: os conflitos armados que se traduziram num cenário de catástrofe social, onde imperava a necessidade de cuidar do Outro. Conclusão: É inegável que os contextos de catástrofes sociais permitiram um campo de atuação que conferiu visibilidade à enfermagem moderna e possibilitou um caminho, traçado no sentido de construir a enfermagem enquanto disciplina.

Abstract Background: Nurses' socio-professional affirmation becomes a reference in contexts of social disasters. Objective: To analyze the relationship between social disasters and the importance of the contribution of some nurses to designing/developing nursing as a profession at the end of the 19th century. Methodology: Although the methodological choices in the validation of the scientific nature of this historical study are based on historiography, the facts narrated and interpreted in this study were contextualized according to the historical period, which allowed the development of new meanings that define unities, totalities, series, and innovative relations within the documentary material itself. Results: Several nurses contributed to the development of nursing as a science. They practiced their profession in contexts with a common denominator: armed conflicts that translated into a scenario of social disaster, where the need to care for others was paramount. Conclusion: Contexts of social disasters unquestionably provided modern nursing with a field of action that brought it visibility and allowed its development as a discipline.

Resumen Contexto: En el contexto de las catástrofes sociales es donde la afirmación socioprofesional del enfermero constituye un referente. Objetivo: Dimensionar la relación entre las catástrofes sociales y la relevancia del trabajo realizado por determinadas enfermeras a finales del siglo XIX, que contribuyeron a la construcción/evolución de la enfermería como profesión. Metodología: Aunque las opciones metodológicas para validar el carácter científico de este estudio histórico se basan en la historiografía, los hechos narrados e interpretados se contextualizaron según el período histórico, lo que permite que surjan nuevos significados que hacen posible definir, en el propio tejido documental, unidades, conjuntos, series, relaciones innovadoras. Resultados: Varias enfermeras contribuyeron a la evolución de la enfermería como ciencia. Vivieron su profesión en contextos con un denominador común, conflictos armados que derivaron en un escenario de catástrofe social, donde primaba la necesidad de cuidar al Otro. Conclusión: Es innegable que los contextos de las catástrofes sociales propiciaron un campo de acción que dio visibilidad a la enfermería moderna y habilitó un camino hacia la construcción de la enfermería como disciplina.

Melhoria da qualidade da água de rios urbanos: novos paradigmas a explorar - Bacia hidrográfica do rio Pinheiros em São Paulo / Improvement of water quality in urban rivers: new paradigms to explore - Pinheiros river basin, São Paulo, Brazil

RESUMO A questão da qualidade da água nos corpos d'água urbanos é complexa, pois passa pelo controle de cargas poluidoras pontuais e difusas. As cargas pontuais ainda são um problema recorrente no Brasil, porém são mais facilmente identificáveis. As cargas difusas não têm um ponto de lançamento específico, o que torna seu controle mais difícil, pois ocorrem principalmente em função da lavagem das superfícies durante as chuvas. Os corpos d'água em áreas urbanas são severamente afetados por tais descargas, que se refletem na qualidade das águas e em seus usos. Nesse contexto, este trabalho apresenta elementos do monitoramento da qualidade da água do rio Pinheiros nos últimos dez anos; analisa os resultados do monitoramento da Companhia Ambiental do Estado de São Paulo (Cetesb); verifica se há relação com resultados do monitoramento diário do projeto Avaliação da Qualidade das Águas do Sistema Pinheiros-Billings com o Protótipo Flotação; e, por fim, avalia o uso de sistema de biorretenção, que apresenta resultados significativos de reduções da carga poluidora do escoamento superficial chegando, em alguns casos, a mais de 90%. Essa solução é uma alternativa para a redução da poluição difusa na bacia do rio Pinheiros.

Abstract The issue of water quality in urban water bodies is complicated, as it involves controlling on point and diffuse pollution loads. Point loads are still a problem in Brazil, although more easily identifiable. Diffuse loads do not have a specific discharge point, which makes their control more difficult, and they happen mainly due to stormwater surface runoff. Urban water bodies are exposed to the diffuse pollution, which affects their water quality and multiple uses. Thus, this article presents the Pinheiros River's water quality monitoring over the last ten years, analyzes the monitoring results of São Paulo State Environmental Company (Cetesb), and verifies the existence of a relationship with these results with the daily monitoring of the project "Avaliação da Qualidade das Águas do Sistema Pinheiros-Billings com o Protótipo Flotação". Finally, it evaluates the use of a bioretention system with average reductions of runoff pollutants loads, in some cases, more than 90%, being an alternative for diffuse pollution control in the Pinheiros River basin.

Psychological well-being of palliative care professionals: Who cares?

BACKGROUND: Traditionally, the psychological well-being of healthcare workers has been taken for granted - it has even been considered a part of the requirements that were demanded of them. When these professionals have experienced suffering and psychological depletion, they have been held accountable for this suffering, adopting an individualistic and reductionist viewpoint focused only on the professional. This approach has become obsolete due to its proven ineffectiveness, especially from an ethics of responsibility and organization viewpoint. CONTEXT: The psychological well-being of the healthcare worker (and its opposites: suffering, exhaustion, and disenchantment) is advantageous to the professional's commitment to the institution, to their work performance, and to their personal life. OBJECTIVE: The objective of this paper is to reflect on the psychological suffering of the palliative care professional. METHOD: We will reflect on the three levels of responsibility that influence such suffering (micro-meso-macro-ethical; worker-environment-institution). RESULTS: We will propose a global strategy for the care of psychological well-being supported by scientific evidence and key references. SIGNIFICANCE OF RESULTS: We conclude with some contributions on what we have learned and still have to learn on this topic.

Identification and characterization of a polyurethanase with lipase activity from Serratia liquefaciens isolated from cold raw cow's milk.

Lipases are associated with food spoilage and are also used in various biotechnological applications. In this study, we sought to purify, identify, and characterize a lipase from S. liquefaciens isolated from cold raw cow's milk. The lipase partially purified by ultrafiltration and gel filtration showed a specific activity of 2793 U/mg. By zymography, the enzyme presented approximately 65 kDa, and LC-MS/MS allowed the identification of a polyurethanase with a conserved domain of family I.3 lipase. The modeled and validated structure of polyurethanase was able to bind to different fatty acids and urethane by molecular docking. The polyurethanase showed optimum activity at pH 8.0 and 30 °C. In the presence of ions, activity was decreased, except for Ca2+, Mg2+, and Ba2+. Reducing agents did not alter the activity, while amino acid modifiers reduced enzyme activity. It is concluded that polyurethanase with lipase activity represents a potential enzyme for the deterioration of milk and dairy products, as well as a candidate for industrial applications.

Nisin Z Production by Wild Strains of Lactococcus lactis Isolated from Brazilian (Italian Type) Fermented Sausage.

In this study, five bacteriocin-producing Lactococcus lactis strains were identified from different naturally fermented Brazilian sausages. Ion exchange and reversed-phase chromatographies were used to purify the bacteriocins from culture supernatant of the five strains. Mass spectrometry (MALDI-TOF/TOF) showed that the molecular masses of the bactericoins from L. lactis ID1.5, ID3.1, ID8.5, PD4.7, and PR3.1 were 3330.567 Da, 3330.514 Da, 3329.985 Da, 3329.561 Da, and 3329.591 Da, respectively. PCR product sequence analysis confirmed that the structural genes of bacteriocins produced by the five isolates are identical to the lantibiotic nisin Z. Optimal nisin Z production was achieved in tryptone and casein peptone, at pH 6.0 or 6.5. The most favorable temperatures for nisin Z production were 25°C and 30°C, and its production was better under aerobic than anaerobic condition. The type of carbon source appeared to be an important factor for nisin Z production. While sucrose was found to be the most efficient carbon source for nisin Z production by four L. lactis isolates, fructose was the best for one isolate. Lactose was also a good energy source for nisin Z production. Surprisingly, glucose was clearly the poorest carbon source for nisin Z production. The five isolates produced different amounts of the bacteriocin, L. lactis ID1.5 and ID8.5 isolates being the best nisin Z producers. DNA sequence analysis did not reveal any sequence differences in the nisZ and nisF promoter regions that could explain the differences in nisin Z production, suggesting that there should be other factors responsible for differential nisin Z production by the isolates.

Proteomic and phosphoproteomic analyses reveal several events involved in the early stages of bovine herpesvirus 1 infection.

Herpesviruses are predicted to express more than 80 proteins during their infection cycle. The proteins synthesized by the immediate early genes and early genes target signaling pathways in host cells that are essential for the successful initiation of a productive infection and for latency. In this study, proteomic and phosphoproteomic tools showed the occurrence of changes in Madin-Darby bovine kidney cells at the early stage of the infection by bovine herpesvirus 1 (BoHV-1). Proteins that had already been described in the early stage of infection for other herpesviruses but not for BoHV-1 were found. For example, stathmin phosphorylation at the initial stage of infection is described for the first time. In addition, two proteins that had not been described yet in the early stages of herpesvirus infections in general were ribonuclease/angiogenin inhibitor and Rab GDP dissociation inhibitor beta. The biological processes involved in these cellular responses were repair and replication of DNA, splicing, microtubule dynamics, and inflammatory responses. These results reveal pathways that might be used as targets for designing antiviral molecules against BoHV-1 infection.

Extraction and identification of antimicrobial peptides from the Canastra artisanal minas cheese.

Canastra artisanal Minas cheese samples were collected in Minas Gerais - Brazil. The samples were evaluated in order to observe the presence of antimicrobial peptides during 30 days of ripening. Soluble peptides extracted from the cheeses were fractionated by reverse phase liquid chromatography and their fractions evaluated for inhibitory action of E. coli. Fractions containing antimicrobial activity were analyzed by MALDI-TOF/TOF and then peptides were sequenced and identified using MASCOT Daemon coupled with UniProt database. The identified peptides were then validated by SCAFFOLD application. The peptides present in fractions with antimicrobial activity were RPKHPIKHQ, RPKHPIKHQG, RPKHPIKHQGLPQ and RPKHPIKHQGLPQE, HQPHQPLPPT and MHQPHQPLPPT. Peptide sequences PKHPIKHQ, RPKHPIKHQG, RPKHPIKHQGLPQ and RPKHPIKHQGLPQE were originated from αs1-casein and are their fragments belonging to Isracidine, which in turn is a well known antimicrobial peptide. The HQPHQPLPPT and MHQPHQPLPPT peptides were related to ß-casein and were isolated in other studies, but their biological activities are still unknown.

Anti-Hypertensive Peptides Derived from Caseins: Mechanism of Physiological Action, Production Bioprocesses, and Challenges for Food Applications.

This review is focused on the state-of-art of peptides with inhibitory activity towards angiotensin I-converting enzyme (ACE) - thus, with anti-hypertensive potential - derived from enzymatic hydrolysis of caseins. Firstly, molecular characteristics of caseins relevant to a better understanding of this subject were concisely commented. Next, a brief description of the pathophysiology of hypertension was explained, focusing on the ACE role in regulation of blood pressure in human body. Then, casein-derived peptides with ACE inhibitory capacity were specifically addressed. The main in vitro and in vivo bioassays often reported in literature to assess the anti-hypertensive potential of peptides were presented, illustrated with recently published studies, and discussed in terms of advantages and limitations of both approaches. Characteristics related to amino acid composition and sequence of peptides with high ACE-inhibitory potential were also commented. Process parameters of enzymatic hydrolysis (types and origins of casein substrates, types of enzymes, pH, temperature, and times of reactions) were discussed. Patents dealing with casein-derived anti-hypertensive peptides were examined not only in terms of amino acid sequences, but also regarding their novelty claims in hydrolysis process parameters. Finally, some trends, challenges, and opportunities inferred from this literature analysis were commented, emphasizing the importance of this research topic in food products development.

Comparative analysis of constitutive proteome between resistant and susceptible tomato genotypes regarding to late blight.

Late blight is one of the most destructive diseases of the tomato, resulting in substantial economic losses. There is difficulty in controlling this disease, so the molecular characterization of tomato genotypes may help in the selection of higher resistance tomato plants against Phytophthora infestans, late blight's pathogen. The objective was to analyze the differences with regard to the constitutive proteome between the access Vegetable Germplasm Bank (BGH)-2127, resistant genotype, and Santa Clara-susceptible genotype to late blight. Proteomic analysis of leaf samples by two-dimensional electrophoresis (2-DE) followed by identification by mass spectrometry (MALDI TOF/TOF) was performed. Nineteen proteins were identified, which were then related to metabolism and energy, photosynthesis, transcription, stress, and defenses. Approximately 90% of these proteins were more abundant in Santa Clara, a susceptible cultivar. Acidic 26 kDa endochitinase and ribonuclease T2 proteins were more abundant in BGH-2127 access. The enzymatic activity confirmed a greater abundance of chitinase in the BGH-2127 access as compared to the cultivar Santa Clara. Gene expression analyses by real-time PCR demonstrated that the mRNA levels were not correlated with the respective protein levels. Abundance of the acidic 26 kDa endochitinase and ribonuclease T2 proteins in the constitutive proteomes of BGH-2127 may be associated with the answer to the resistance of this access.

Proteomic analysis of the venom of the predatory ant Pachycondyla striata (Hymenoptera: Formicidae).

The ants use their venom for predation, defense, and communication. The venom of these insects is rich in peptides and proteins, and compared with other animal venoms, ant venoms remain poorly explored. The objective of this study was to evaluate the protein content of the venom in the Ponerinae ant Pachycondyla striata. Venom samples were collected by manual gland reservoir dissection, and samples were submitted to two-dimensional gel electrophoresis and separation by ion-exchange and reverse-phase high-performance liquid chromatography followed by mass spectrometry using tanden matrix-assisted laser desorption/ionization with time-of-flight (MALDI-TOF/TOF) mass spectrometry and electrospray ionization-quadrupole with time-of-flight (ESI-Q/TOF) mass spectrometry for obtaining amino acid sequence. Spectra obtained were searched against the NCBInr and SwissProt database. Additional analysis was performed using PEAKS Studio 7.0 (Sequencing de novo). The venom of P. striata has a complex mixture of proteins from which 43 were identified. Within the identified proteins are classical venom proteins (phospholipase A, hyaluronidase, and aminopeptidase N), allergenic proteins (different venom allergens), and bioactive peptides (U10-ctenitoxin Pn1a). Venom allergens are among the most expressed proteins, suggesting that P. striata venom has high allergenic potential. This study discusses the possible functions of the proteins identified in the venom of P. striata.

Purification and characterization of two new cell-bound bioactive compounds produced by wild Lactococcus lactis strain.

Novel compounds and innovative methods are required considering that antibiotic resistance has reached a crisis point. In the study, two cell-bound antimicrobial compounds produced by Lactococcus lactis ID1.5 were isolated and partially characterized. Following purification by cationic exchange and a solid-phase C18 column, antimicrobial activity was recovered after three runs of RPC using 60% (v/v) and 100% (v/v) of 2-propanol for elution, suggesting that more than one antimicrobial compound were produced by L. lactis ID1.5, which were in this study called compounds AI and AII. The mass spectrum of AI and AII showed major intensity ions at m/z 1070.05 and 955.9 Da, respectively. The compound AI showed a spectrum of antimicrobial activity mainly against L. lactis species, while the organisms most sensitive to compound AII were Bacillus subtilis, Listeria innocua, Streptococcus pneumoniae and Pseudomonas aeruginosa. The antimicrobial activity of both compounds was suppressed by treatment with Tween 80. Nevertheless, both compounds showed high stability to heat and proteases treatments. The isolated compounds, AI and AII, showed distinct properties from other antimicrobial substances already reported as produced by L. lactis, and have a significant inhibitory effect against two clinically important respiratory pathogens.