Variation In Protein Profiles Research Articles

Sodium Dodecyl Sulphate-polyacrylamide Gel Electrophoresis (SDS-PAGE) Characterization of Protein Fractions in Ten Peanut Bunch Type Varieties

Aim: To isolate these fractions, a progressive centrifugation technique was employed, utilizing water, sodium chloride, phosphate buffer, and alcohol. Place and Duration of Study: A study conducted in department of biochemistry Junagadh Agricultural University, Junagadh for analysis seeds of the bunch-type groundnut were supplied by the Main Oilseeds Research Station in Junagadh during 2020-2022. The Methodology Used: Using SDS-PAGE separation method, four key protein fractions: albumin, prolamin, globulin, and glutelin were analyzed. Results: The results revealed the highest albumin and globulin contents ranging from 15.05-20.81% and 69.91-77.99%, respectively. The globulin fraction was found to be significantly higher in proportion compared to the other three fractions. Glutelin and prolamin were found to be very low percentage-wise in bunch varieties but it was found to be in the range of 1.49-3.42 % in glutelin as well as prolamins were found to be in the range of 1.5-3.95%. The SDS-PAGE analysis of the ten-bunch type varieties revealed significant variations in protein profiles, as evidenced by the total number of bands and their respective molecular weight-retention factor (MW-Rf) values. The albumin and globulin fractions exhibited the highest MW-Rf values collectively, whereas glutelin and prolamin demonstrated minimal banding patterns. Conclusion: This study concludes that the observed qualitative and quantitative differences in seed protein profiles are invaluable for varietal identification.

An in silico Approach For Identification of Potential Therapeutic Targets For Cancer Treatment From Celastrus hindsii Benth

Celastrus hindsii Benth., a medicinal plant celebrated for its traditionally medicinal and practically therapeutic properties, has been used for generations in Vietnam to support the treatment of ulcers, tumors, and inflammation. The difference between several phenotypes, primarily identified as Broad Leaf (BL) and Narrow Leaf (NL), has been clarified by convincing scientific evidence through our previous proteomics study, which also revealed several bioactive proteins and peptides. Therefore, based on the findings, this study further investigated their therapeutic properties using a bioinformatics tool (BLASTP) and analyzing literature data. The results showed the distinguished variations in protein profile between the NL and BL proteomes and revealed five significant proteins with therapeutic properties. Of these, three proteins can have anti-tumor and anti-inflammatory activity and have been proven effective in cancer treatment. Therefore, C. hindsii, particularly the BL phenotype with elevated levels of therapeutic proteins, could be a promising plant candidate for future intensive research and applications for cancer treatment.

Parasitic infections, hematological and biochemical parameters suggest appropriate health status of wild coati populations in anthropic Atlantic Forest remnants.

Coatis are hosts of a great diversity of parasites, that due to anthropic pressures in forest fragments, like changes in landscapes and ecosystems, can influence the dynamics and physiological responses to those parasite infections, affecting the animal's health and fitness. This is the first study about health parameters and parasitic infections of wild coati (Nasua nasua) populations in the Atlantic Forest (Pernambuco Center of Endemism). The following hypotheses were evaluated: (i) infections and co-infections by gastrointestinal parasites and ectoparasites can generate changes in the health parameters of coatis such as the body condition score (BCS), packed cell volume (PCV), leukogram, and serum protein profile; (ii) biological aspects (sex and age) or fragment they inhabit, can influence changes in the health parameters (BCS, PCV, leukogram and serum protein profile). Were studied 55 free-living coatis in three anthropized forest remnants in the Metropolitan Region of Recife. After chemical containment, the animals were submitted to physical examination and collection of biological samples (blood, feces, and ectoparasites). On the physical examination, 23.6% of coatis had a low BCS and 5.4% were overweighted. Amblyomma spp. ticks were found in 83.6% of the animals of all studied remnants, A. sculptumAmblyomma sculptum in 12.7% and A. ovale in 1.8%. Regarding gastrointestinal parasites, Ancylostoma sp. was the most prevalent (80.4%) and most animals (66.7%) had co-infection with Ancylostoma sp. and Capillaria sp., Strongyloides sp., Acanthocephala, Cestoda, and Coccidia. The 76.5% of the coatis presented co-infections with Ancylostoma spp. + Amblyomma spp. Principal coordinates analyses (PCoA) scores of health parameters were used as dependent variables and fragment, sex, age, Ancylostoma sp. infection, gastrointestinal parasites co-infection, Amblyomma spp. infestation and co-infection of Ancylostoma sp. + Amblyomma spp. as a predictor variable in the linear models. Parasites did not influence the PCV of the individuals, but a decrease was evident in adult animals. Variations in protein profile, neutrophils, and lymphocytes, without leaving the normal range for the species, but WBC were predicted by age group, and infections by Ancylostoma or Amblyomma spp., but not their co-infections. The free-living coati populations of the anthropized remnants in the Atlantic Forest of northeastern Brazil proved to be healthy and seem to be adapted to face the challenges of anthropization and parasitic infections.

The effect of thermal pasteurization, freeze-drying, and gamma irradiation on donor human milk

The availability of donor human milk (DHM) is currently limited by the volumes that can be thermally pasteurized and kept in long-term cold storage. This study assesses the application of freeze-drying followed by low-dose gamma irradiation of DHM for simplified, safe long-term storage. Solid-phase microextraction (SPME) GC–MS, SDS and native PAGE gel electrophoresis demonstrated that the overall changes in volatile and protein profiles in Holder pasteurized and freeze-dried DHM was negligible compared to the natural variations in DHM. Freeze-dried DHM samples (moisture < 2.2 %) processed with 2 kGy gamma irradiation did not show any significant lipid oxidation end-products and variation in protein profile. Therefore, freeze-drying followed by in-packaging gamma irradiation could be a safe method for pasteurization, convenient storage and delivery of DHM at ambient temperature. These methods may generate a means to create a reserve stock of DHM for emergencies and humanitarian aid.

Protein profiling of chicken breast muscles from different slaughter houses in relation to meat quality

Introduction: Chicken meat is a source of protein in the human diet. Protein content and values define the quality of chicken meat. This research aimed to analyze variations of protein profiles in chicken breast muscles from different slaughtering houses by using proteomic strategies. Methods: Total proteins of chicken breast muscles from three different slaughtering houses (Sample A, Sample B and Sample C) were extracted and quantified by using Bradford assay. Then, the proteins were separated by SDS-PAGE to monitor the quality of extracted proteins. Protein profiles in different samples were compared by 2D-GE analysis. Results: The most highly expressed protein band was located between the molecular size of 37-50 kD in all samples and it was expected to be betaactin. While resolved in 2D-PAGE, differences in protein expression were observed between samples. There were three spots expressed with highest intensity in Sample B compared to others. The protein spot detected at pH 5.28 and the size range between 50- 75 kD was predicted to be NADPH-cytochrome P450 reductase (CPR), at pH 6.45 and molecular weight between 37-50 kD was expected to be creatine kinase M-type (M-CK) while at pH 6.78 and molecular size nearly 25 kD was expected to be αB-crystallin. Conclusions: It can be mentioned that these proteins could play a vital role in mechanisms that contribute to the poor quality of chicken meat.

Effect of Alternative Pasteurization Techniques on Human Milk's Bioactive Proteins.

Human milk (HM) feeding leads to improved outcome for preterm infants. When mother's milk is unavailable, pasteurized donor HM (DHM) is the recommended alternative over formula. The Holder pasteurization (HoP) method is universally performed in HM banks; however, it is known to impair several functional HM components. The aim of this study was to compare the efficacy of HoP with 2 innovative processing methods (high-temperature short-time [HTST] pasteurization and high-pressure processing [HPP]) in preserving some bioactive HM protein components. HM samples from donors of the Bologna HM bank were collected and divided into 4 subsamples: 1 was kept raw, and each of the others was processed using a different technique (HoP, HTST, and HPP at 600 MPa for 3 minutes). Total protein content, secretory immunoglobulin A (sIgA), and lactoferrin contents were compared. Both HM lactoferrin and sIgA content were negatively affected, but to a different extent, by each method: sIgA was preserved by HTST, with only HPP leading to a significant reduction (-38.8%); lactoferrin content was strongly reduced by HoP (-87.5%) and HTST (-83.5%), and preserved by HPP. Variations in protein profile were seen for all processing methods, being more relevant for HoP, followed by HTST and, finally, by HPP. All the 3 methods lowered the untreated HM microbial counts to undetectable levels, in accordance with national guidelines. Both HTST and HPP better preserved the original HM protein profile, compared to HoP. They, however, affected differently some bioactive HM components involved in immune response and antibacterial activity.

Smell of Infection: A Novel, Noninvasive Method for Detection of Fish Excretory-Secretory Proteins.

Chemical signals are produced by aquatic organisms following predatory attacks or perturbations such as parasitic infection. Ectoparasites feeding on fish hosts are likely to cause release of similar alarm cues into the environment due to the stress, wounding, and immune response stimulated upon infection. Alarm cues are often released in the form of proteins, antimicrobial peptides, and immunoglobulins that provide important insights into bodily function and infection status. Here we outline a noninvasive method to identify potential chemical cues associated with infection in fish by extracting, purifying, and characterizing proteins from water samples from cultured fish. Gel free proteomic methods were deemed the most suitable for protein detection in saline water samples. It was confirmed that teleost proteins can be characterized from water and that variation in protein profiles could be detected between infected and uninfected individuals and fish and parasite only water samples. Our novel assay provides a noninvasive method for assessing the health condition of both wild and farmed aquatic organisms. Similar to environmental DNA monitoring methods, these proteomic techniques could provide an important tool in applied ecology and aquatic biology.

Protein Levels and Microstructural Changes in Localized Regions of Early Cartilage Degeneration Compared with Adjacent Intact Cartilage.

It was hypothesized that the respective protein profiles of bovine cartilage from sites of localized mild to moderate (GI to GII) degeneration versus adjacent sites of intact tissue would vary in accordance with the tissue microstructural changes associated with a pre-osteoarthritic state. A total of 15 bovine patellae were obtained for this study. Paired samples of tissue were collected from the lateral region of each patella. If the patella contained a site of degeneration, a paired tissue set involved taking one sample each from the degenerated site and the intact tissue adjacent to it. Sufficient tissue was collected to facilitate 2 arms of investigation: microstructural imaging and proteome analysis. The microstructural analysis used a bespoke tissue preparation technique imaged with differential interference contrast optical microscopy to assess fibrillar scale destructuring and underlying bone spicule formation. An iTRAQ-based proteome analysis was performed using liquid chromatography-tandem mass spectrometry to identify the differential levels of proteins across the intact and degenerated cartilage and further, the results were validated with multiple reaction monitoring assay. In the healthy cartilage pairs, there was no significant variation in protein profiles between 2 adjacent sample sites. In pairs of tissue that contained a sample of GI/GII tissue, there were both significant microstructural changes as well as the difference in abundance levels of 24 proteins. From the known functions of the 24 proteins, found to be strongly aligned with the specific microstructural changes observed, a unique "proteins ensemble" involved in the initiation and progression of early cartilage degeneration is proposed.

Levels of human proteins in plasma associated with acute paediatric malaria

BackgroundThe intimate interaction between the pathophysiology of the human host and the biology of the Plasmodium falciparum parasite results in a wide spectrum of disease outcomes in malaria. Development of severe disease is associated with a progressively augmented imbalance in pro- and anti-inflammatory responses to high parasite loads and sequestration of parasitized erythrocytes. Although these phenomena collectively constitute common denominators for the wide variety of discrete severe malaria manifestations, the mechanistic rationales behind discrepancies in outcome are poorly understood. Exploration of the human pathophysiological response by variations in protein profiles in plasma presents an excellent opportunity to increase the understanding. This is ultimately required for better prediction, prevention and treatment of malaria, which is essential for ongoing elimination and eradication efforts.ResultsAn affinity proteomics approach was used to analyse 541 paediatric plasma samples collected from community controls and patients with mild or severe malaria in Rwanda. Protein profiles were generated with an antibody-based suspension bead array containing 255 antibodies targetting 115 human proteins. Here, 57 proteins were identified with significantly altered levels (adjusted p-values < 0.001) in patients with malaria compared to controls. From these, the 27 most significant proteins (adjusted p-values < 10−14) were selected for a stringent analysis approach. Here, 24 proteins showed elevated levels in malaria patients and included proteins involved in acute inflammatory response as well as cell adhesion. The remaining three proteins, also implicated in immune regulation and cellular adhesivity, displayed lower abundance in malaria patients. In addition, 37 proteins (adjusted p-values < 0.05) were identified with increased levels in patients with severe compared to mild malaria. This set includes, proteins involved in tissue remodelling and erythrocyte membrane proteins. Collectively, this approach has been successfully used to identify proteins both with known and unknown association with different stages of malaria.ConclusionIn this study, a high-throughput affinity proteomics approach was used to find protein profiles in plasma linked to P. falciparum infection and malaria disease progression. The proteins presented herein are mainly involved in inflammatory response, cellular adhesion and as constituents of erythrocyte membrane. These findings have a great potential to provide increased conceptual understanding of host-parasite interaction and malaria pathogenesis.

Biological and Proteolytic Variation in the Venom of Crotalus scutulatus scutulatus from Mexico.

Rattlesnake venoms may be classified according to the presence/absence and relative abundance of the neurotoxic phospholipases As (PLAs), such as Mojave toxin, and snake venom metalloproteinases (SVMPs). In Mexico, studies to determine venom variation in Mojave Rattlesnakes (Crotalus scutulatus scutulatus) are limited and little is known about the biological and proteolytic activities in this species. Tissue (34) and venom (29) samples were obtained from C. s. scutulatus from different locations within their distribution in Mexico. Mojave toxin detection was carried out at the genomic (by PCR) and protein (by ELISA) levels for all tissue and venom samples. Biological activity was tested on representative venoms by measuring LD and hemorrhagic activity. To determine the approximate amount of SVMPs, 15 venoms were separated by RP-HPLC and variation in protein profile and proteolytic activity was evaluated by SDS-PAGE (n = 28) and Hide Powder Azure proteolytic analysis (n = 27). Three types of venom were identified in Mexico which is comparable to the intraspecific venom diversity observed in the Sonoran Desert of Arizona, USA: Venom Type A (∼Type II), with Mojave toxin, highly toxic, lacking hemorrhagic activity, and with scarce proteolytic activity; Type B (∼Type I), without Mojave toxin, less toxic than Type A, highly hemorrhagic and proteolytic; and Type A + B, containing Mojave toxin, as toxic as venom Type A, variable in hemorrhagic activity and with intermediate proteolytic activity. We also detected a positive correlation between SVMP abundance and hemorrhagic and proteolytic activities. Although more sampling is necessary, our results suggest that venoms containing Mojave toxin and venom lacking this toxin are distributed in the northwest and southeast portions of the distribution in Mexico, respectively, while an intergradation in the middle of both zones is present.

High C-reactive proteins levels, rheumatoid anemia, alpha-1 globulin deficiency, and hypergammaglobulinemia in rheumatoid arthritis patients from yaounde, Cameroon

Background: Rheumatoid arthritis (RA) is an autoantibodies-mediated disease affecting 0.5%–1% of the population worldwide. The present study was designed to investigate variations of protein profiles and hematological parameters of patients in Yaounde Central Hospital. Methods: The study was conducted from January to November 2017. RA cases were recruited based on the identification of established diagnosis from registers of patients of participating rheumatologists. Healthy volunteers visiting the hospital were also included as a control population. All participants were aged 15 years and above. Whole blood collected from each participant was assayed for hematological parameters. In addition, serum from each participant was assayed for the level of C-reactive proteins (CRPs) using a plate agglutination technique whereas plasma was used for protein profile through a conventional electrophoretic migration of proteins on cellulose acetate gel. Results: Overall, 22 RA patients and 10 controls were recruited and consisted, respectively, in 5 males and 17 females and 2 males and 8 females. Our study revealed that 59.09% (13/22) of patients presented high levels of CRP compared to 10% in the control group. The mean value of red blood cells was 4.38 × 106/μl and 4.698 × 106/μl, respectively, in the RA and the control groups. The mean hemoglobin value was significantly reduced in the RA group (11.07 mg/dl vs. 12.73 mg/dl, P = 0.0192), thus revealing anemia in patients. Nearly 54.5% of patients presented hypoalbuminemia compared to 20% in the control group (P = 0, 1241). A deficiency of α1-globulin was observed in 54.5% of patients while that of control group was 30%. Furthermore, 72% of the patient presented hypergammaglobulinemia compared to 30% in the control group (P = 0.0494). Conclusion: This study revealed that RA patients present a normochromic and normocytic anemia, a decreased albumin and alpha 1 globulin, a significant elevation of gamma-globulins compared to the control population.

Quantitative proteomic analysis of host responses triggered by Mycobacterium tuberculosis infection in human macrophage cells.

Macrophages are primary host of Mycobacterium tuberculosis (M.tb) and the central effector of in vivo innate immune responses against bacteria. Though the interaction between macrophages and mycobacteria has been widely investigated, the molecular mechanisms of M.tb pathogenesis in macrophages are still not clear. In this work, we investigated the altered protein expression profiles of macrophages after virulent H37Rv strain and avirulent H37Ra strain infection by tandem mass tag-based quantitative proteomics. Among 6762 identified proteins of macrophages, the expression levels of 235 proteins were significantly altered, which is supposed to be related to the infection of different strains. By bioinformatics analysis at systems level, we found that these proteins are mainly involved in the biological process of apoptosis, blood coagulation, oxidative phosphorylation, and others. The enormous variation in protein profiles between macrophages infected with H37Ra and H37Rv suggests the existence of four different immunity mechanisms that decide the fates of macrophages and M.tb. These data may provide a better understanding of M.tb pathogenesis within the host, which contributes to the prevention and clinical treatment of tuberculosis.

Endurance assessment of Eichhornia crassipes (Mart.) Solms, in heavy metal contaminated site–A case study

AbstractIn this study, the ability to hyper accumulate heavy metals from contaminated site by water hyacinth (Eichhornia crassipes) was monitored. The heavy metal-contaminated habitat was validated by X-ray diffraction and energy dispersive X-ray elemental spectrometry analysis of the soil samples. Heavy metal contamination in soil and water; accumulation in foliar, root and bulb tissue samples were determined by atomic absorption spectroscopy and were monitored as a function of accumulation in different tissues. Significant differences were recorded in the bioaccumulation capability of heavy metals by different tissue. Discrete variation in protein profile of leaves and high expression of alcohol dehydrogenase (ADH), peroxidase (POX) and altered regulation of esterase (EST) in root tissue was observed in contaminated site grown Eichhornia. The high metal accumulation efficiency of water hyacinth due to the biomass production suggests this species as reliable organic biomarker for heavy metal contamination.

English

Korean ginseng is a traditional medicine that is widely used in Korea. In this study, a proteomic approach was used to investigate variations in Korean ginseng isolates that are associated with ecologic and geographic differences. Ginseng samples were collected from four geographically isolated locations in Korea: North gyeonggi, Gochang, Geumsan and Kanghwa. Two-dimensional gel electrophoresis (2-DE) and peptide fingerprinting of tryptic digests by mass spectrometry (MALDI-TOF) revealed primary ginseng root region-specific variations in protein profiles in these distinct areas. Thirty seven (37) major proteins that are common to the main root of ginseng at all four geographic sites and six proteins that are specific to the main root of a local ginseng (Kanghwa) were identified. Most of the major common proteins identified could be classified into the following functional categories: (i) stress response; (ii) transcription and translation; (iii) nucleotide metabolism; (iv) plant hormone response; (v) signal transduction; (vi) protein degradation; (vii) protein destination and storage; and (viii) unassigned. The results show that Korean ginseng species can be distinguished on the basis of classical proteomics. Key words: Panax ginseng C.A. Meyer, 2-DE, peptide fingerprinting, classical proteomics.

Genomic Analysis of Storage Protein Deficiency in Genetically Related Lines of Common Bean (Phaseolus vulgaris).

A series of genetically related lines of common bean (Phaseolus vulgaris L.) integrate a progressive deficiency in major storage proteins, the 7S globulin phaseolin and lectins. SARC1 integrates a lectin-like protein, arcelin-1 from a wild common bean accession. SMARC1N-PN1 is deficient in major lectins, including erythroagglutinating phytohemagglutinin (PHA-E) but not α-amylase inhibitor, and incorporates also a deficiency in phaseolin. SMARC1-PN1 is intermediate and shares the phaseolin deficiency. Sanilac is the parental background. To understand the genomic basis for variations in protein profiles previously determined by proteomics, the genotypes were submitted to short-fragment genome sequencing using an Illumina HiSeq 2000/2500 platform. Reads were aligned to reference sequences and subjected to de novo assembly. The results of the analyses identified polymorphisms responsible for the lack of specific storage proteins, as well as those associated with large differences in storage protein expression. SMARC1N-PN1 lacks the lectin genes pha-E and lec4-B17, and has the pseudogene pdlec1 in place of the functional pha-L gene. While the α-phaseolin gene appears absent, an approximately 20-fold decrease in β-phaseolin accumulation is associated with a single nucleotide polymorphism converting a G-box to an ACGT motif in the proximal promoter. Among residual lectins compensating for storage protein deficiency, mannose lectin FRIL and α-amylase inhibitor 1 genes are uniquely present in SMARC1N-PN1. An approximately 50-fold increase in α-amylase inhibitor like protein accumulation is associated with multiple polymorphisms introducing up to eight potential positive cis-regulatory elements in the proximal promoter specific to SMARC1N-PN1. An approximately 7-fold increase in accumulation of 11S globulin legumin is not associated with variation in proximal promoter sequence, suggesting that the identity of individual proteins involved in proteome rebalancing might also be determined at the translational level.

Common and metal-specific proteomic responses to cadmium and zinc in the metal tolerant ericoid mycorrhizal fungus Oidiodendron maius Zn.

Although adaptive metal tolerance may arise in fungal populations in polluted soils, the mechanisms underlying metal-specific tolerance are poorly understood. Comparative proteomics is a powerful tool to identify variation in protein profiles caused by changing environmental conditions, and was used to investigate protein accumulation in a metal tolerant isolate of the ericoid mycorrhizal fungus Oidiodendron maius exposed to zinc and cadmium. Two-dimensional gel electrophoresis and shotgun proteomics followed by mass spectrometry lead to the identification of common and metal-specific proteins and pathways. Proteins selectively induced by cadmium exposure were molecular chaperons of the Hsp90 family, cytoskeletal proteins and components of the translation machinery. Zinc significantly up-regulated metabolic pathways related to energy production and carbohydrates metabolism, likely mirroring zinc adaptation of this fungal isolate. Common proteins induced by the two metal ions were the antioxidant enzyme Cu/Zn superoxide dismutase and ubiquitin. In mycelia exposed to zinc and cadmium, both proteomic techniques also identified agmatinase, an enzyme involved in polyamine biosynthesis. This novel finding suggests that, like plants, polyamines may have important functions in response to abiotic environmental stress in fungi. Genetic evidence also suggests that the biosynthesis of polyamines via an alternative metabolic pathway may be widespread in fungi.

Unconditioned commercial embryo culture media contain a large variety of non-declared proteins: a comprehensive proteomics analysis.

Which non-declared proteins (proteins not listed on the composition list of the product data sheet) are present in unconditioned commercial embryo culture media? A total of 110 non-declared proteins were identified in unconditioned media and between 6 and 8 of these were quantifiable and therefore represent the majority of the total protein in the media samples. There are no data in the literature on what non-declared proteins are present in unconditioned (fresh media in which no embryos have been cultured) commercial embryo media. The following eight commercial embryo culture media were included in this study: G-1 PLUS and G-2 PLUS G5 Series from Vitrolife, Sydney IVF Cleavage Medium and Sydney IVF Blastocyst Medium from Cook Medical and EmbryoAssist, BlastAssist, Sequential Cleav and Sequential Blast from ORIGIO. Two batches were analyzed from each of the Sydney IVF media and one batch from each of the other media. All embryo culture media are supplemented by the manufacturers with purified human serum albumin (HSA 5 mg/ml). The purified HSA (HSA-solution from Vitrolife) and the recombinant human albumin supplement (G-MM from Vitrolife) were also analyzed. For protein quantification, media samples were in-solution digested with trypsin and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). For in-depth protein identification, media were albumin depleted, dialyzed and concentrated before sodium dodecyl sulfate polyacrylamide gel electrophoresis. The gel was cut into 14 slices followed by in-gel trypsin digestion, and analysis by LC-MS/MS. Proteins were further investigated using gene ontology (GO) terms analysis. Using advanced mass spectrometry and high confidence criteria for accepting proteins (P < 0.01), a total of 110 proteins other than HSA were identified. The average HSA content was found to be 94% (92-97%) of total protein. Other individual proteins accounted for up to 4.7% of the total protein. Analysis of purified HSA strongly suggests that these non-declared proteins are introduced to the media when the albumin is added. GO analysis showed that many of these proteins have roles in defence pathways, for example 18 were associated with the innate immune response and 17 with inflammatory responses. Eight proteins have been reported previously as secreted embryo proteins. For six of the commercial embryo culture media only one batch was analyzed. However, this does not affect the overall conclusions. The results showed that the HSA added to IVF media contained many other proteins and that the amount varies from batch to batch. These variations in protein profiles are problematic when attempting to identify proteins derived from the embryos. Therefore, when studying the embryo secretome and analyzing conditioned media with the aim of finding potential biomarkers that can distinguish normal and abnormal embryo development, it is important that the medium used in the experimental and control groups is from the same batch. Furthermore, the proteins present in unconditioned media could potentially influence embryonic development, gestation age, birthweight and perhaps have subsequent effects on health of the offspring. The study was supported by the Danish Agency for Science, Technology and Innovation. Research at the Fertility Clinic, Aarhus University Hospital is supported by an unrestricted grant from Merck Sharp & Dohme Corp and Ferring. The authors declare no conflicts of interest.

Efeito da temperatura das amostras de leite na concentração de cálcio solúvel e de beta-caseína: interferência no teste de estabilidade frente ao etanol

O objetivo deste trabalho foi avaliar a variação do perfil proteico e do cálcio solúvel na coagulação do leite pelo etanol nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Amostras de leite de 61 animais foram avaliadas quanto à estabilidade ao etanol nas concentrações de 66 a 92% (v/v) nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Três amostras, após 24 horas de armazenamento a 4ºC, foram ultracentrifugadas em quadruplicata (40.000 x g) a 4ºC e a 20ºC, respectivamente, por 60 minutos. Em seguida, o sobrenadante foi retirado e submetido à análise do cálcio solúvel pela técnica via úmida (digestão nitroperclórica) e leitura em espectrofotômetro de absorção atômica. O perfil proteico foi analisado pela técnica de eletroforese capilar empregando kit específico para determinação proteica. Os resultados mostraram uma correlação positiva entre o aumento da temperatura das amostras e a estabilidade do leite frente às diferentes concentrações de etanol. A porcentagem de cálcio solúvel no sobrenadante após ultracentrifugação foi maior nas amostras tratadas a 4ºC (P&lt;0,05). As amostras ultracentrifugadas na temperatura de 4ºC apresentaram quantidades superiores de β-caseína no sobrenadante em comparação com as amostras tratadas a 20ºC. O abaixamento da temperatura favoreceu a migração da β-caseína e do cálcio coloidal para a fase solúvel do leite, o que possivelmente favoreceu o aumento da instabilidade das amostras no teste do etanol. Os resultados sugerem que a temperatura ideal para a realização de teste de estabilidade do leite frente ao etanol deveria ser de 21ºC.

Differential proteomics of dehydration and rehydration in bryophytes: evidence towards a common desiccation tolerance mechanism

All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms.

Quantitative Assessment of Regulatory Proteins in Blood as Markers of Radiation Effects in the Late Period After Occupational Exposure

The objective of this research was quantitative assessment of serum and membrane regulatory proteins in blood from nuclear workers as markers of radiation-induced alterations in immune homeostasis in the late period after protracted exposure of nuclear workers with different doses. The effector and regulatory lymphocytes were measured using a flow cytofluorometer in workers from the main facilities of the Mayak PA (aged ∼60 y up to 80 y) in the late period after combined exposure to external gamma-rays and internal alpha-radiation from incorporated 239Pu. The control group included non-occupationally exposed members of the Ozyorsk population matched by gender and age to the group of Mayak workers. Thirty serum proteins involved in regulation of immune homeostasis, such as growth factors, multifunctional interleukins, pro- and anti-inflammatory cytokines, and their receptors, were measured using ELISA in blood serum specimens from the Radiobiology Human Tissue Repository. The dosimetry estimates were obtained using Doses-2005. The correlation analysis revealed a statistically significant direct relationship of T-killers and plutonium body burden and a decreasing level of T-helpers with accumulated external dose in exposed individuals. There were differences in expression of membrane markers in young regulatory cells (double null T-lymphocytes, NKT-lymphocytes, regulatory T-cells, and an increase of activated forms of T-lymphocytes), which indicated an active role of regulatory cells in maintaining immune homeostasis in terms of protracted exposure. The assessment of regulatory proteins in blood indicated that growth factors (EGF, TGF-β1, PDGF), multifunctional interleukins (IL-17A, IL-18), and pro-inflammatory cytokines (IL-1β and INF-γ) could be potential markers of radiation-induced alterations in protein status. An imbalance of pro- and antiinflammatory proteins in blood and variations of protein profiles at the lower exposure levels (gamma-ray dose <1 Gy, plutonium body burden <0.74 kBq) in the late period after protracted exposure were less pronounced than at the higher exposure levels, which was probably explained by compensatory-adaptive responses in the late period among senile individuals with polypathology.