Changes In Protein Patterns Research Articles

Effect of Naphthalene acetic acid and date palm pollen extraction on gene expression on fruit of Hillawi date palm cultivar during growth and ripening

This study was carried out on fruit of Hillawi date palm cultivar which pollinated by Khukri Adi andGhunami Akhder cultivars to study the effect of NAA 25 ppm and pollen extraction of Khukri Adi andGhunami Akhder 2g/L which were sprayed at 30 and 45 day after pollination on gene expression bystudying the changes in protein pattern during fruit development and ripening by gel electrophoresesmethod with sodium dodecyle sulfate (SDS-PAGE). The result showed there were many protein bandwhich were different in molecular weight and numbers on the gel with changes on fruit growth stages andtreatments, These bands of protein were similar in early stage of growth but different in the period of13,15 weeks after pollination, this difference may be correlated with ripening date. Our results showedthat we could consider the protein pattern as gene expression markers for fruit and linking them with adifferent growth and ripening stage and explanation of metaxenia phenomenon.

Methyl parathion (50% ec) induced changes in protein and DNA banding patterns in the fish Channa punctatus (bloch)

The acute toxicity tests for 24, 48, 72 and 96 h to the fish Channa punctatus were conducted with an organophosphate pesticide methyl parathion 50% EC in both static and continuous flow-through methods. The LC 50 values obtained for static test were 0.6661, 0.6473, 0.6200 and 0.5949 ppm respectively for static and 0.5736, 0.5592, 0.5364 and 0.521 ppm respectively for continuous flow-through method. The 1/10 th of static 96 h LC 50 value was taken as sub-lethal concentration and the fish were exposed to both lethal and sub-lethal concentrations for 8 days and the vital tissue of the fish viz ., gill, liver, brain, muscle and kidney were isolated and analyzed for study of changes in protein and DNA banding patterns. The Rm values were derived from the electrophoretogram along with the marker. The changes in Rm values indicate that marked variations were taken place in both the protein and DNA bands. The changes in lethal exposure were more prominent compared to sub-lethal exposure. These changes were attributed to the intoxication of the test toxicant methyl parathion. The results obtained were discussed with the available literature.

PD30-09 TRANSURETHRAL RESECTION OR INCISION OF THE PROSTATE AFTER RENAL TRANSPLANTATION: IS THERE A SAFE TIME FOR THE PROCEDURE?

improvements in graft survival following kidney transplantation. However, there are no non-invasive diagnostic parameters available that enable early and reliable detection of acute rejection. The objective of our study was to establish a urinary protein profile suitable to determine rejectionspecific changes following kidney transplantation and to predict early acute rejection at a postoperative stage. Furthermore, we focused on identification of candidate proteins for the use as biomarkers in clinical practice for diagnosis of acute rejection. METHODS: Urine samples of 116 kidney recipients were included. Rejection was proven by biopsy (n1⁄458) and stable transplant function was monitored for at least 2 years (n1⁄458). Postoperative urine samples were collected between 3rd and 10th day following transplantation. Urinary protein profiles were analyzed by SELDI-TOF-MS. Multiplex-Fluorescence-2DE and Peptide Mass Fingerprinting were used for identification of candidate proteins. Urinary concentration of candidate proteins was validated by ELISA. RESULTS: A protein signature including 4 masses differentiated acute rejection from stable transplant patients at the postoperative stage with 73% sensitivity and 88% specificity. Alpha-1-microglobulin (A1MG) and Haptoglobin (Hp) were identified as putative biomarkers for acute rejection. Protein levels were significantly higher in postoperative urine samples from patients with upcoming rejection than in stable transplant patients (A1MG: 29.13 mg/ml vs. 22.06 mg/ml, p1⁄40.001; Hp: 628.34 ng/ml vs. 248.57 ng/ml, p1⁄40.003). The combination of both proteins enabled the diagnosis of early rejection with 85% sensitivity and 80% specificity. CONCLUSIONS: Protein profiling using mass spectrometry is suitable for non-invasive detection of rejection-specific changes following kidney transplantation. A specific protein profile enables prediction of early acute allograft rejection in the immediate postoperative period. A1MG and Hp appear to be reliable rejection biomarkers. Further analyses have to show if early diagnosis is possible for patients with allograft rejection that occurs several month after transplantation by analyzing changes in urine protein pattern in regular intervals.

Potentiality of Garlic and/or Green Tea Aqueous Extracts in Attenuating Pesticides Induced Toxicity in Albino Rats

Aim: The present work aimed to evaluate the effect of each of green tea (GT) and garlic (Ga) aqueous extract and their combination on pesticide Duracine induced changes in plasma levels of some indices of oxidative stress; and in protein electrophoretic patterns, in albino rats. Study Design: Forty rats were divided into 8 groups 5rats each and treated orally for 14 days as follows: G1 (control);-G 2 (200mg GT per animal);G 3 (1ml of 50% of garlic):G4 (200mg green tea per animal and 1ml of 50% of garlic); G 5 20mg /Kg bwt [equivelant 1/14 LD50 of Duracine] ,G6 (green tea and Duracine):G7 (garlic and Duracine):G8 (green tea, garlic and Duracine). Plasma activities of acetylcholinesterase (AChE), glutathione-Stransferase (GST) and glutathione reductase (GR) activities and levels of malondialdehyde (MDA) and total reduced glutathione (GSH) were analyzed using standard methods. Results: Duracine intoxication induced significant decrease in plasma AChE, GST and Original Research Article article........ Article Annual Research & Review in Biology, 4(8): 1290-1306, 2014 1291 GR. It elevated plasma MDA, decreased plasma total GSH, as well as, changes in plasma protein electrophoretic pattern. Individual supplementation with GT induced slight improvement in AchE, GSH and MDA. On the other hand, supplementation with aqueous extract of garlic induced significant (P<0.05) enhancement in the level of GSH in the intoxicated animals. Combined supplementation with GT+Ga produced significant (P<0.05) counteracting effect in MDA,GSH,GST and GR, as well as, counteracting changes in plasma protein pattern was not remarkable this may be due to plasma protein half life. Conclusion: It seems that, combined supplementation of green tea and garlic improve the oxidative stress parameters but needa more time to improve other toxicity markers.

Changes in the serum protein electrophoretic pattern in lambs during the first month of life

Studies of the changes in serum protein pattern in the neonatal period in animals are still limited. Therefore, the objective of this study was to evaluate the changes in the concentrations of serum protein fractions in 7 clinically healthy merino lambs (4 males, 3 females) during their first month of life. The first blood sampling was performed before the colostrum intake and then at 1, 2, 7, 14 and 30 days of age. Blood serum was analysed for total serum protein concentrations and for the relative and absolute values of serum protein fractions - albumin, alpha1- (α1), alpha2- (α2), beta- (β), and gamma- (γ) globulins. The results showed a significant effect of age on the serum total protein concentrations and for all the protein fractions. The concentrations of total proteins and γ-globulins increased significantly 1 day after the colostrum intake (P&lt; 0.001); then decreased gradually till the end of the 1st month. An opposite trend was observed in the concentrations of albumin. A significant decrease of values was found 1 day after the intake of colostrum (P&lt; 0.001) with a subsequent progressive increase of values till the end of the evaluated period. The relative concentrations of α1-globulins significantly decreased during the first month of life (P&lt; 0.001). On the other hand, the relative values of α2- and β-globulins increased significantly from birth till the end of the monitored period (P&lt; 0.001). Our results suggest that the serum protein electrophoretic pattern in growing lambs is significantly influenced by the age of the evaluated animal, and this should be taken into consideration when interpreting the serum protein profile. Our findings extend existing knowledge about significant changes in the protein profile associated with the physiological adaptation process in the neonatal period in young animals.

The in vitro effects of CoCl2 as ethylene synthesis inhibitor on PI based protein pattern of potato plant (Solanum tuberosum L.)

The effect of CoCl2 as an ethylene synthesis inhibitor on changes of protein pattern was investigated in potato (Solanum tuberosum L.) plants cultivar White Desiree. In vitro grown plants were subjected to MS medium containing 0, 5, 10, 15, 20, 30, 40 and 60 mg/l CoCl2 for 4 weeks. Different concentrations of CoCl2 showed significant effect on the total soluble proteins. Among different concentrations of CoCl2, using 20 mg/l CoCl2 was the best concentration to inhibit ethylene formation and induce potato plant growth. Application of CoCl2 in the culture medium changed the total protein as well as SDS-PAGE and Iso-electric Point Electrophoresis (PI) patterns. Protein pattern in potato tuber did not show any detectable changes.

돼지에서 발정 주기 동안 자궁액 내의 단백질 패턴의 변화

An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184～187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.

소의 난소주기 동안 과립막세포와 황체조직에서 단백질 패턴의 변화

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89～93 in ovulation phase were transferrin, and spot No.94～98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC- E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

High-content cytometry and transcriptomic biomarker profiling of human B-cell activation

Primary antibody deficiencies represent the most prevalent, although very heterogeneous, group of inborn immunodeficiencies, with a puzzling complexity of cellular and molecular processes involved in disease pathogenesis. We aimed to study in detail the kinetics of CD40 ligand/IL-21-induced B-cell differentiation to define new biomarker sets for further research into primary antibody deficiencies. We applied high-content screening methods to monitor B-cell activation on the cellular (chip cytometry) and transcriptomic (RNA microarray) levels. The complete activation process, including stepwise changes in protein and RNA expression patterns, entry into the cell cycle, proliferation and expression of activation-induced cytidine deaminase (AID), DNA repair enzymes, and post-class-switch expression of IgA and IgG, was successfully monitored during in vitro differentiation. We identified a number of unknown pathways engaged during B-cell activation, such as CXCL9/CXCL10 secretion by B cells. Finally, we evaluated a deduced set of biomarkers on a group of 18 patients with putative or proved intrinsic B-cell defects recruited from the European Society for Immunodeficiencies database and successfully predicted 2 AID defects and 1 DNA repair defect. Complete absence of class-switched B cells was a sensitive predictor of AID deficiency and should be further evaluated as a diagnostic biomarker. The biomarkers found in this study could be used to further study the complex process of B-cell activation and to understand conditions that lead to the development of primary antibody deficiencies.

Effects of a highly resistant rice starch and pre-incubation temperatures on the physicochemical properties of surimi gel from grass carp (Ctenopharyn Odon Idellus)

The effects of a specific rice starch (SRS), containing highly resistant starch (RSIII), on gel properties of grass carp (Ctenopharyngodon idella) and the influence of five levels of SRS (0%, 2%, 4%, 6%, and 8%w/w) on gel physicochemical properties at three different pre-incubation temperatures (4°C, 25°C, and 40°C) were investigated. Gels with increasing amounts of SRS addition showed lower expressible water contents and cooking loss values than did control gels. SDS gel electrophoresis revealed no changes in protein patterns, regardless of different SRS-added levels at the same pre-incubation temperature, but an evident decrease in the MHC when the pre-incubation temperature increased. The texture properties, colour attributes, SEM were optimal in the treatments containing 4%w/w SRS at the pre-incubation temperature 25°C. Thus, the optimum SRS addition level and pre-incubation temperature are proposed to be 4%w/w and 25°C, respectively.

From Gene Engineering to Gene Modulation and Manipulation: Can We Prevent or Detect Gene Doping in Sports?

During the last 2 decades, progress in deciphering the human gene map as well as the discovery of specific defective genes encoding particular proteins in some serious human diseases have resulted in attempts to treat sick patients with gene therapy. There has been considerable focus on human recombinant proteins which were gene-engineered and produced in vitro (insulin, growth hormone, insulin-like growth factor-1, erythropoietin). Unfortunately, these substances and methods also became improper tools for unscrupulous athletes. Biomedical research has focused on the possible direct insertion of gene material into the body, in order to replace some defective genes in vivo and/or to promote long-lasting endogenous synthesis of deficient proteins. Theoretically, diabetes, anaemia, muscular dystrophies, immune deficiency, cardiovascular diseases and numerous other illnesses could benefit from such innovative biomedical research, though much work remains to be done. Considering recent findings linking specific genotypes and physical performance, it is tempting to submit the young athletic population to genetic screening or, alternatively, to artificial gene expression modulation. Much research is already being conducted in order to achieve a safe transfer of genetic material to humans. This is of critical importance since uncontrolled production of the specifically coded protein, with serious secondary adverse effects (polycythaemia, acute cardiovascular problems, cancer, etc.), could occur. Other unpredictable reactions (immunogenicity of vectors or DNA-vector complex, autoimmune anaemia, production of wild genetic material) also remain possible at the individual level. Some new substances (myostatin blockers or anti-myostatin antibodies), although not gene material, might represent a useful and well-tolerated treatment to prevent progression of muscular dystrophies. Similarly, other molecules, in the roles of gene or metabolic activators [5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR), GW1516], might concomitantly improve endurance exercise capacity in ischaemic conditions but also in normal conditions. Undoubtedly, some athletes will attempt to take advantage of these new molecules to increase strength or endurance. Antidoping laboratories are improving detection methods. These are based both on direct identification of new substances or their metabolites and on indirect evaluation of changes in gene, protein or metabolite patterns (genomics, proteomics or metabolomics).

Hepatic DNA Damage and Abnormality in Serum Protein Pattern Due to Long Term Use of Tramadol in Rats

Tramadol is a widely used analgesic that stimulates the μ opioid receptor and inhibits serotonin and noradrenalin reuptake. In this study, we investigate the effect of chronic administration of 200 and 400mg of tramal for 15,30 ,45 days followed by withdrawal periods(w15) on protein activity in male rats as manifested by changes in electrophoretic serum protein patterns and gene expression manifested by DNA damage, measured by Comet Assay . Materials and methods: 100 male Wistar rats (100-150 g) were included and divided into three groups, control group (n = 20), Tramal(I) group (n = 40), received the drug orally at doses of 200mg/kg/day for 15,30,45 days of the study, respectively(10 rats for each subgroup).TR45 group was followed by 15 withdrawal period(W15) ,Tramal(II) group (n = 40), received the drug orally at doses of400mg/kg/day for 15,30,45 days of the study, respectively. TR45 group was followed by 15 withdrawal period (W15). Results: Results exhibited major changes in the protein pattern which included changes in the molecular weight of the control bands and the relative percentage of protein fraction as well as the total number of bands, as a result of disappearance of some original bands and appearance of other new one. Serum protein fraction revealed an increase in total number of protein fractions being in Tr200mg (14 and 16 bands), whileTR400mg revealed a decrease in total number of bands (except TR15 which exhibited an increase in bands (14). The changes were observed all over the treated groups as well as in the withdrawal groups In this study, the alkaline version of the comet assay has been used to determine the effect of tramal administration (200 and 400mg/Kg) on peroxide-initiated free radical-mediated DNA damage in rat liver cells. Indeed, levels of strand breaks in rat liver cell exposed to tramal400mg/Kg were significantly higher than in cells exposed to 200mg/Kg, especially after a long administration period (TR45 days). The intensity of the comet tail relative to the head reflects the number of DNA breaks. The rates of tailed cells detected by the comet assay increased significantly when the rats were exposed to 200 and 400mg/kg of tramal compared with control (however, the tail length did not differ significantly between the same groups). The intensity of the comet tail relative to the head reflects the number of DNA breaks. Conclusions: Our findings pointed out the risk of increased lipid peroxidation, hepatic DNA damage and abnormality in serum protein pattern due to long term use of tramadol, although opioids are reported to be effective in pain management, their toxic effects should be kept in mind.

인공위액조건과 예열처리에 따른 GM 콩 도입단백질(CP4EPSPS)의 소화성 평가

Gastrointestinal digestibility of new proteins inserted in the food supply is a significant parameter for assessing the safety of GM foods based on the assumption that digestive stability is undesirable. In this study, we performed in vitro digestion of CP4EPSPS, a new protein, expressed in genetically modified (GM) soybean in order to evaluate its digestibility in three different ratios of simulated gastric fluid with preheating. Ratios of GM soybean to simulated gastric fluid were 2:2, 2.5:1.5, and 1.5:2 and preheating was conducted at 100oC for 5 min. Electrophoresis and Western blotting were used to confirm changes in soybean protein patterns and CP4EPSPS gene expression after in vitro digestion. At ratios in which the amount of gastric fluid was equal to (2:2) or relatively higher than that of soybean (1.5:2), no CP4EPSPS (47.4 kDa) protein was detected after 15 seconds of simulated gastric fluid incubation, the earliest time interval evaluated. However, when the ratio of GM soybean to gastric fluid was 2.5:1.5, CP4EPSPS was detected in 5 min and gradually decreased according to time. After preheating, no CP4EPSPS protein was detected after 15 seconds under all conditions. From these results, we concluded that the digestibility of CP4EPSPS in simulated gastric fluid increased upon preheating. Accordingly, we suggest that it is important to account for the ratio of gastric fluid to GM food in in vitro digestibility assessment models of GM food.

Cadmium and zinc induced similar changes in protein and glycoprotein patterns in tobacco (nicotiana tabacum l.) seedlings and plants

The effects of 10 μmol L-1 and 15 μmol L-1 cadmium (Cd), a nonessential toxic element and 25 μmol L-1 and 50 μmol L-1 zinc (Zn), an essential micronutrient, on proteins and glycoproteins of Nicotiana tabacum L. seedlings and plants were investigated after exposure to each metal alone or to their combinations. Changes in only few polypeptides related to heavy metal treatments were observed in tobacco seedlings and leaves of adult plants, while the greatest change in total soluble protein pattern was observed in plant roots. Differences between control and treated tobacco tissues were more pronounced in the glycoprotein pattern, which was analysed by application of different lectins. The majority of the detected glycoproteins in leaves and roots of adult plants can be considered as a result of enhanced glycosylation due to heavy metal stress. The difference in glycoproteins between Cd and Zn application on tobacco seedlings and adult plants could not be determined since enhanced glycosylation was noticed after treatment with either metal alone or in combination. Therefore, it can be concluded that both metals induced N- and Oglycosylation as a result of changed environmental conditions.

Growth Inhibition by miR-519 via Multiple p21-Inducing Pathways

The microRNA miR-519 robustly inhibits cell proliferation, in turn triggering senescence and decreasing tumor growth. However, the molecular mediators of miR-519-elicited growth inhibition are unknown. Here, we systematically investigated the influence of miR-519 on gene expression profiles leading to growth cessation in HeLa human cervical carcinoma cells. By analyzing miR-519-triggered changes in protein and mRNA expression patterns and by identifying mRNAs associated with biotinylated miR-519, we uncovered two prominent subsets of miR-519-regulated mRNAs. One subset of miR-519 target mRNAs encoded DNA maintenance proteins (including DUT1, EXO1, RPA2, and POLE4); miR-519 repressed their expression and increased DNA damage, in turn raising the levels of the cyclin-dependent kinase (cdk) inhibitor p21. The other subset of miR-519 target mRNAs encoded proteins that control intracellular calcium levels (notably, ATP2C1 and ORAI1); their downregulation by miR-519 aberrantly elevated levels of cytosolic [Ca(2+)] storage in HeLa cells, similarly increasing p21 levels in a manner dependent on the Ca(2+)-activated kinases CaMKII and GSK3β. The rises in levels of DNA damage, the Ca(2+) concentration, and p21 levels stimulated an autophagic phenotype in HeLa and other human carcinoma cell lines. As a consequence, ATP levels increased, and the level of activity of the AMP-activated protein kinase (AMPK) declined, further contributing to the elevation in the abundance of p21. Our results indicate that miR-519 promotes DNA damage, alters Ca(2+) homeostasis, and enhances energy production; together, these processes elevate the expression level of p21, promoting growth inhibition and cell survival.

Retardation of post-mortem changes of freshwater prawn (Macrobrachium rosenbergii) stored in ice by legume seed extracts

Meat quality of freshwater prawn (Macrobrachium rosenbergii) treated with soybean and bambara groundnut extracts at different concentrations was monitored during 10days of iced storage. During storage, the control sample (without treatment) had a higher pH, TCA-soluble peptide content, heat soluble collagen content, proteolytic activities and psychrophilic bacterial count than did samples treated with soybean and bambara groundnut extracts. Conversely, shear force value and likeness scores of the control sample decreased (p<0.05), more likely associated with softening of muscle. The decrease in myosin heavy chain in the control sample was found after 6days of storage. However, no changes in protein patterns of samples treated with soybean extracts at 2.5mg/mL were found after 10days of storage. Therefore, the injections of legume seed extracts, especially soybean extract, at a sufficient concentration, could be a means to retard muscle softening and maintain the qualities of freshwater prawn during iced storage.

Changes in apoplast protein pattern suggest an early role of cell wall structure remodelling in flagellin-triggered basal immunity

The leaf apoplast is a dynamic compartment in contact with plant pathogenic bacteria after infection. Among the very first interaction events is the receptor-mediated perception of bacterial surface molecules such as flagellin or other conserved microbe-associated molecular patterns (MAMPs). Apoplast proteins likely play a role in basal resistance (BR) or pattern-triggered immunity (PTI). Here, a proteomic approach was carried out on water soluble - potentially the most mobile - apoplast proteins from flagellin-treated tobacco (Nicotiana tabacum) leaves. As the quickness of BR/PTI seems crucial for its efficacy, samples were taken as early as 2.5 and 7 h post inoculation. Proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and identified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Forty-nine different proteins from 28 protein spots changed in their density compared to the water-inoculated control. Eleven protein spots appeared de novo in response to EBR induction. There are glycohydrolases and redox-active proteins besides pathogenesis-related proteins among them, predicting plant cell wall structural modifications and more direct antimicrobial effectors as earliest changes related to BR/PTI.

Changes in protein pattern during different developmental stages of somatic embryos in chickpea

Mature embryonic axes were used for chickpea (Cicer arietinum L.) regeneration via somatic embryogenesis. Qualitative and quantitative estimation of protein profile during somatic embryogenesis by SDS-PAGE and densitometric analysis showed differential expression of various storage proteins at different stages of somatic embryo development, which was compared with the profile of developing seeds. Total protein content in somatic embryos of chickpea increased from globular stage [2.9 &mu;g mg-1(f.m.)] to cotyledonary stage [4.8 &mu;g mg-1(f.m.)] and then started decreasing during onset of maturation and germination [up to 1.5 &mu;g mg-1(f.m.)]. Differential expression of seed storage proteins, late embryogenesis abundant (LEA) proteins and proteins related with stress response were documented at different stages of somatic embryogenesis. Germinating somatic embryos showed degradation products of several seed storage proteins and the appearance of new polypeptides (76.8, 67.6, 49.9 and 34.2 kDa), which were absent during differentiation of somatic embryos. A low molecular mass (17.7 kDa) polypeptide was uniformly present during all stages of somatic embryogenesis and it may belong to a group of stress-related proteins. This study describes the expression of true seed storage proteins like legumin, vicilin, convicilin and their subunits at different stages of somatic embryogenesis, which may serve as excellent markers for embryogenic pathway of regeneration in chickpea.

Proteomic characterization by 2-DE in bovine serum and whey from healthy and mastitis affected farm animals

Acute phase proteins (APP) have been identified in whey and sera from healthy and mastitis cows through the proteomic analysis using two-dimensional electrophoresis (2-DE) coupled with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Although normal and mastitis serum samples show relatively similar protein composition, marked differences in expression levels and patterns can be observed. Conversely, normal and mastitis whey showed a very different composition, likely due to extravasation of blood proteins to the mammary gland. Different isoforms from the most abundant protein in milk, casein, were detected in both normal and mastitis whey. Other proteins, such as lactotransferrin, were only detected in the inflamed animal samples. Immunoglobulins showed different patterns but not increased levels in the inflamed whey. Also, many cellular proteins in mastitis cow's whey, that were absent from healthy cow's milk. They are responsible for the great change in composition between normal and mastitis whey, especially those which exert a biological function related to immune defense. Data collected in this work are of interest for gaining information about physiological changes in protein patterns in different fluids and, the correspondent modifications as result of an acute phase process in farm. This article is part of a Special Issue entitled: Proteomics: The clinical link.

Microparticle enlargement and altered surface proteins after air decompression are associated with inflammatory vascular injuries

Studies in a murine model have shown that decompression stress triggers a progressive elevation in the number of circulating annexin V-coated microparticles derived from leukocytes, erythrocytes, platelets, and endothelial cells. We noted that some particles appeared to be larger than anticipated, and size continued to increase for ≥24 h postdecompression. These observations led to the hypothesis that inert gas bubbles caused the enlargement and particle size could be reduced by hydrostatic pressure. After demonstrating pressure-induced particle size reduction, we hypothesized that annexin V-positive particle changes associated with decompression contributed to their proinflammatory potential. Intravenous injection of naive mice with particles isolated from decompressed mice, but not control mice, caused intravascular neutrophil activation; perivascular neutrophil sequestration and tissue injuries were documented as elevations of vascular permeability and activated caspase-3. These changes were not observed if mice were injected with particles that had been subjected to hydrostatic recompression or particles that had been emulsified by incubation with polyethylene glycol telomere B surfactant. Hydrostatic pressure and surfactant incubation also altered the pattern of proteins expressed on the surface of particles. We conclude that proinflammatory events and vascular damage are due to enlargement of annexin V-coated particles and/or changes in surface marker protein pattern associated with provocative decompression. Injection of annexin V-coated particles from decompressed mice will recapitulate the pathophysiological vascular changes observed following decompression stress.