scholarly journals Quantitative Proteome Reveals Variation in the Condition Factor of Sea Urchin Strongylocentrotus nudus during the Fishing Season Using an iTRAQ-based Approach

Marine Drugs ◽  
2019 ◽  
Vol 17 (7) ◽  
pp. 397
Author(s):  
Wen-Hui Shang ◽  
Jia-Run Han ◽  
Jia-Nan Yan ◽  
Yi-Nan Du ◽  
Yun-Sheng Xu ◽  
...  
Keyword(s):  
Protein Content ◽  
Sea Urchin ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Condition Factor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gonad Index ◽  
Multifunctional Protein ◽  
Strongylocentrotus Nudus

To investigate the variation in the condition factor of the sea urchin Strongylocentrotus nudus (S. nudus), gonads were collected in May (MAY), June (JUN), and July (JUL), at the beginning (AUG-b) and end of August (AUG-e). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) detection of the gonads revealed an obvious enhancement of the band at about 37 kDa from July, which was identified as transforming growth factor-beta-induced protein ig-h3 (TGFBI) by nanoLC-ESI-MS/MS. Gonadal proteins were identified by isobaric tagging for relative and absolute quantitation (iTRAQ), and regulation of the identified proteins in pairs of the collected groups was observed. A total of 174 differentially expressed proteins (DEPs) were identified. Seven of the DEPs showed significant correlations with both the gonad index (GI) and protein content. These correlations included 6-phosphogluconate dehydrogenase, decarboxylating isoform X2 (6PGD), CAD protein, myoferlin isoform X8, ribosomal protein L36 (RL36), isocitrate dehydrogenase [NADP], mitochondrial isoform X2 (IDH), multifunctional protein ADE2 isoform X3, sperm-activating peptides (SAPs) and aldehyde dehydrogenase, and mitochondrial (ALDH). However, TGFBI had no correlation with gonad index (GI) or protein content. 6PGD, IDH, multifunctional protein ADE2 isoform X3, and ALDH were shown to interact with each other and might play key roles in changing the condition factor of S. nudus gonads.

X-linked inhibitor of apoptosis protein (XIAP) inhibition in systemic sclerosis (SSc)

2021 ◽  
pp. annrheumdis-2020-219822
Author(s):  
Christina Bergmann ◽  
Ludwig Hallenberger ◽  
Sara Chenguiti Fakhouri ◽  
Benita Merlevede ◽  
Amelie Brandt ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Transforming Growth Factor Beta ◽  
Topoisomerase I ◽  
Transforming Growth Factor ◽  
Inhibitor Of Apoptosis ◽  
Dependent Manner ◽  
Cultured Fibroblasts ◽  
Multifunctional Protein ◽  
Inhibitor Of Apoptosis Protein ◽  
Apoptosis Protein

ObjectiveX-linked inhibitor of apoptosis protein (XIAP) is a multifunctional protein with important functions in apoptosis, cellular differentiation and cytoskeletal organisation and is emerging as potential target for the treatment of various cancers. The aim of the current study was to investigate the role of XIAP in the pathogenesis of systemic sclerosis (SSc).MethodsThe expression of XIAP in human skin samples of patients with SSc and chronic graft versus host disease (cGvHD) and healthy individuals was analysed by quantitative PCR, immunofluorescence (IF) and western blot. XIAP was inactivated by siRNA-mediated knockdown and pharmacological inhibition. The effects of XIAP inactivation were analysed in cultured fibroblasts and in the fibrosis models bleomycin-induced and topoisomerase-I-(topoI)-induced fibrosis and in Wnt10b-transgenic mice.ResultsThe expression of XIAP, but not of other inhibitor of apoptosis protein family members, was increased in fibroblasts in SSc and sclerodermatous cGvHD. Transforming growth factor beta (TGF-β) induced the expression of XIAP in a SMAD3-dependent manner. Inactivation of XIAP reduced WNT-induced fibroblast activation and collagen release. Inhibition of XIAP also ameliorated fibrosis induced by bleomycin, topoI and overexpression of Wnt10b in well-tolerated doses. The profibrotic effects of XIAP were mediated via WNT/β-catenin signalling. Inactivation of XIAP reduces binding of β-catenin to TCF to in a TLE-dependent manner to block WNT/β-catenin-dependent transcription.ConclusionsOur data characterise XIAP as a novel link between two core pathways of fibrosis. XIAP is overexpressed in SSc and cGvHD in a TGF-β/SMAD3-dependent manner and in turn amplifies the profibrotic effects of WNT/β-catenin signalling on fibroblasts via transducin-like enhancer of split 3. Targeted inactivation of XIAP inhibits the aberrant activation of fibroblasts in murine models of SSc.

15 SPERM STORAGE IN FEMALE REPRODUCTIVE TRACT: STUDY OF MOLECULES INVOLVED

2015 ◽  
Vol 27 (1) ◽  
pp. 100
Author(s):  
C. Riou ◽  
A. Gargaros ◽  
G. Harichaux ◽  
A. Brionne ◽  
J. Gautron ◽  
...  
Keyword(s):  
Protein Content ◽  
Quantitative Methods ◽  
Reproductive Tract ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Protein ◽  
Sperm Storage ◽  
Female Reproductive Tract ◽  
Storage Duration ◽  
Uterine Fluid

Because of prolonged sperm storage in their oviduct, domestic hens can produce fertile eggs for up to 3 weeks following a single AI. The oviduct secretions may have an effect on sperm survival, but its composition during fertilization is unknown. In the present study, we compared the proteomic content of uterine fluid collected from two lines of hens divergent by their duration of fertility period (DFP), which defined sperm-storage duration. The first line displays a shorter period of sperm storage (10 days, line DFP–), whereas the second displays a longer period of sperm storage (21 days, DFP+). The aim was to identify proteins or peptides that may be involved in spermatozoa survival. Uterine fluid was collected 10 h after oviposition either before (n = 5/line) or 24 h after (n = 5/line) AI. Samples were pooled by condition: DFP+ before AI, DFP+ after AI, DFP– before AI, and DFP– after AI. Bottom-up approach using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nano LC-MS/MS was performed (3 replicates). Data were matched against the NCBInr database (2014) using Mascot, and identifications were validated by the peptide and protein Prophet algorithm using Scaffold software. To determine the differences in protein expression, spectral counting and XIC quantitative methods were employed using Scaffold Q+ (P < 0.05, ratio > 2). Two proteins were up-regulated and one was down-regulated in oviductal secretion of both lines in response to AI. However, AI induced a significantly different abundance between protein content of DFP– and DFP+ fluids. A panel of 8 proteins, included one DFP+-specific protein, was more abundant in DFP+ line than in DFP–. Only one protein was less abundant in DFP+ line than in DFP–. In conclusion, the presence of sperm in the genital tract induced quantitative differences of the protein content of the uterine fluid in DFP– and DFP+ hen lines. These differences imply proteins that are known as male proteins (sperm, seminal plasma, testis). Analysis of sperm protein modifications after storage will help us to understand the functional implication of these candidates.

Purification and partial characterization of a mitogenic protein released from preadipocytes of massively obese subjects

1990 ◽  
Vol 68 (4) ◽  
pp. 764-768 ◽  
Author(s):  
Daniel A. K. Roncari ◽  
Coral E. Thompson
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Radioreceptor Assay ◽  
Massive Obesity ◽  
Epidermal Growth ◽  
Obese Subjects ◽  
High Degree

A protein released into the culture medium by omental preadipocytes of massively obese persons, which stimulates the replication of rat perirenal preadipocytes, has been purified to a high degree. By gel filtration chromatography, the molecular mass of the mitogenic protein was ~66 000 daltons (Da), while on sodium dodecyl sulfate – polyacrylamide gel electrophoresis, two subunits were obtained, relative masses (Mr) of ~31 000 and ~35 000. The isoelectric point of the ~66 000 Da entity was 5.6 ± 0.2. By specific radioreceptor assay, the purified protein was related to epidermal growth factor and transforming growth factor alpha. It was not related to insulin-like growth factors I and II by radioimmunoassay and radioreceptor assay. We propose that the ~66 000 Mr protein, and other mitogenic proteins released by preadipocytes from massively obese persons, act through paracrine–autocrine mechanisms and may play a role in the development of the hyperplasia of enlarged fat cells characteristic of massive corpulence.Key words: preadipocyte, massive obesity, mitogenic, paracrine.

Effect of sodium azide on Bambara groundnut (Vigna subterranean (L.) verdc.) as revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (sds-page)

2021 ◽  
Vol 20 (1) ◽  
pp. 183-194
Author(s):  
Odunayo Joseph Olawuyi ◽  
Juliet Ese Naworu ◽  
Roseline Tolulope Feyisola
Keyword(s):  
Protein Content ◽  
Sodium Azide ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Lethal Effect ◽  
Bambara Groundnut ◽  
International Institute ◽  
Yield Traits ◽  
North East ◽  
Sds Page

This study investigated the mutagenic effects of Sodium Azide (NaN3) on the agromorphological and protein content of eight Bambara groundnut genotypes. The seeds of six genotypes; TVSu-86, TVSu-91, TVSu-186, TVSu-235, TVSu-242, TVSu-350 were collected fromthe International Institute of Tropical Agriculture (IITA) and two landraces from Abia State and Enugu State North East, Nigeria local markets. The seeds were treated with five concentrations: 0.00%(control), 0.01%, 0.03%, 0.05% and 0.07% of NaN3 after pre-soakingfor 6hrs in distilled water and sown in pots arranged in a Complete Randomized Design with three replicates. There was reduction in germination percentage and growth characters as concentrations of NaN3 increases. Early flowering was recorded at 37 days mutated with 0.07% of NaN3 compared to control which flowered late at 42 days. NaN3 (0.07%) caused lethal effect on Abia and Enugu landraces. There was no significant (P>0.05) difference in yield traits among mutants and control. Mutant seeds significantly (P<0.05) increased protein content (19.12%) at 0.05% of NaN3 compared to control (18.5%). The number of seeds (0.99), seed yield (0.89) and pod yield (0.96) strongly correlated with seeds per pod (0.85). The SDS-PAGE revealed the presence of polypeptide bands in mutants compared to control. TVSu-235 and TVSu-350 genotypes had higher tolerance and yield traits to 0.01% concentration of NaN3, thuscould be further improved in subsequent breeding. Keywords: Bambara groundnut, Sodium azide, SDS-PAGE, polypeptide bands.

Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin

1996 ◽  
Vol 134 (4) ◽  
pp. 481-489 ◽  
Author(s):  
James R McFarlane ◽  
Lynda M Foulds ◽  
Angelique Pisciotta ◽  
David M Robertson ◽  
David M de Kretser
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Biological Fluids ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Male Rat ◽  
Tween 20 ◽  
Rat Serum ◽  
Level 3

McFarlane JR, Foulds LM, Pisciotta A, Robertson DM, de Kretser DM. Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin. Eur J Endocrinol 1996;134:481–9. ISSN 0804–4643 Activin, a dimer of the β-subunits of inhibin, is a member of the transforming growth factor beta (TGF-β) superfamily of growth factors and has a widespread range of actions in a variety of tissues. The investigation of the physiology of activin action has been facilitated in recent years by the availability of immunoassays in addition to bioassays. Follistatin has been shown to bind to activin with a high affinity and therefore interferes in both radioimmunoassays and enzyme-linked immunosorbent assays (ELISAs). In this study we examined the effect of various surfactants and 1.4-dioxane on the measurement of activin in the presence of follistatin by radioimmunoassay. The addition of a combination of sodium deoxycholate, Tween 20 and sodium dodecyl sulphate removed the interference of follistatin in the radioimmunoassay. The measured content of activin in male rat serum, human male serum, human female serum and bovine follicular fluid rose from 3.29 to 4.15, < 0.48 to 2.87, 2.42 to 4.17 and 30.9 to 85.6 ng/ml, respectively, when assayed in the presence of the dissociating reagents. It was unclear whether the altered potencies were due to a dissociation of the follistatin/activin complex rather than the exposure of the epitope on activin recognized by the antiserum. Serum concentrations of activin were lower than those found in testicular cytosols, and after castration no change in serum activin levels was observed, suggesting that the testis does not contribute significantly to circulating activin levels. The use of the dissociating reagents in the radioimmunoassay will enable studies to be carried out that more accurately measure the activin content of various biological fluids, and thus lead to a greater understanding of the physiology of this growth factor. James McFarlane, Institute of Reproduction and Development, Level 3, Block E, Monash Medical Centre, 246 Clayton Road, Clayton, Victoria 3168, Australia

Hormonally regulated proteins in cultured human fetal lung: analysis by two-dimensional gel electrophoresis

1990 ◽  
Vol 259 (4) ◽  
pp. L283-L293
Author(s):  
M. W. Odom ◽  
R. Ertsey ◽  
P. L. Ballard
Keyword(s):  
Transforming Growth Factor Beta ◽  
Gel Electrophoresis ◽  
Transforming Growth Factor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Fetal Lung ◽  
Gamma Interferon ◽  
Computer Assisted ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Human Fetal Lung

We used high-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) to identify hormonally regulated proteins in cultured human fetal lung. Proteins labeled with [35S]methionine were separated by 2-D PAGE, and fluorograms were analyzed by computer-assisted analysis of densitometric scans. Dexamethasone (10 nM) and gamma-interferon (10 ng/ml) induced (2- to 22-fold vs. control) distinct sets of proteins (comprising approximately 2% of approximately 1,000 resolved proteins). Treatment with forskolin (10 microM) plus 3-isobutyl-1-methylxanthine (IBMX, 100 microM), which increases intracellular adenosine 3',53'-cyclic monophosphate (cAMP), induced both unique proteins and several proteins induced by dexamethasone. One protein (Mr 40,000, pI 4.4) was induced only with combined dexamethasone and cAMP treatment. Dexamethasone repressed four proteins, but inhibition was not observed with other hormones. Some of the regulated proteins were enriched in either fibroblasts or type II cells isolated from lung explants. We found no proteins that were consistently regulated by triiodothyronine (T3) (2 nM) or transforming growth factor-beta (10 ng/ml). Additionally, none of the hormonal treatments substantially altered the rate of methionine incorporation into total protein. Thus we have identified separate subsets of proteins that are regulated by glucocorticoids, gamma-interferon, and cAMP; these proteins may be important mediators of hormonal effects in the developing fetal lung.

scholarly journals TGF-β prevents the denervation-induced reduction of bone formation and promotes the bone regeneration through inhibiting ubiquitin-proteasome pathway

2019 ◽  
Vol 39 (5) ◽  
Author(s):  
Zhen Yu ◽  
Ye Li ◽  
Yining Wang ◽  
Yuting Chen ◽  
Mengfan Wu ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Regeneration ◽  
Transforming Growth Factor Beta ◽  
Bone Growth ◽  
Transforming Growth Factor ◽  
Multifunctional Protein ◽  
Ubiquitin Proteasome Pathway ◽  
Ubiquitin Proteasome ◽  
Proteasome Pathway ◽  
Tgf Β1

Abstract Background: Transforming growth factor beta (TGF-β) can stimulate osteogenesis as a multifunctional protein. The present study was to explore if TGF-β can prevent denervation-induced reduction of bone formation. Materials & methods: The 6-week-old male mice were treated with recombinant human TGF-β1 (rhTGF-β1). Bone formation, endochondral bone growth rates, and gene expression of osteoblast markers were measured in the skeletal tissue by real-time PCR. Results: RhTGF-β1 treatment prevented the denervation-induced decrease in bone formation rates, endochondral growth, and expression of Cbfa1/Runx2 (runt-related transcription factor 2), Ostecalcin (OC), and ColIA1. TGF-β1 partially inhibited the denervation-induced ubiquitination of Cbfa1/Runx2 in mouse cancellous bones via ubiquitin-proteasome pathway. Conclusion: TGF-β prevents denervation-induced reduction of bone formation and promotes the bone regeneration through inhibiting ubiquitin-proteasome pathway at least partially.

scholarly journals Isolation and characterization of the vitelline layer of sea urchin eggs.

1977 ◽  
Vol 75 (2) ◽  
pp. 410-421 ◽  
Author(s):  
C G Glabe ◽  
V D Vacquier
Keyword(s):  
Sea Urchin ◽  
External Surface ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Internal Surface ◽  
Single Amino Acid ◽  
Sea Urchin Eggs ◽  
Isolation And Characterization ◽  
Triton X

The vitelline layers (VLs) of unfertilized sea urchin eggs were isolated by homogenization in a hypotonic medium containing Triton X-100 and EDTA. The surface topography of the VL is not changed by isolation. The thickness of the isolated VLs (300-400 A) is greater than that reported for VLs on intact eggs (100-200 A). Sperm adhere to the isolated VLs. When both internal and external VL surfaces are accessible to sperm, the sperm attach only to the external surface, suggesting that the external surface may carry sperm receptor proteins not present on the internal surface. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis shows that isolated VLs are composed of numerous proteins ranging from greater than 213,000 to 25,000 daltons. Lactoperoxidase-catalyzed 125I-iodination of unfertilized eggs labels two high molecular weight bands that stain faintly for carbohydrate. VLs are 90% protein and 3.5% carbohydrate. No predominance of a single amino acid or class of amino acids was found. Carbohydrate analysis yields fucose, mannose, galactose, glucose, xylose, glucosamine, galactosamine, and sialic acid. Controls for purity indicate that isolated VLs contain 2% protein of cytoplasmic origin and no more than 2.5% egg jelly.

scholarly journals Protein Changes in Peach Seeds during Chilling Are Not Associated with Breaking Dormancy

1994 ◽  
Vol 119 (1) ◽  
pp. 131-135 ◽  
Author(s):  
Ahmed Mahhou ◽  
Frank G. Dennis
Keyword(s):  
Protein Content ◽  
Prunus Persica ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Germination Capacity ◽  
Protein Profiles ◽  
Soluble Protein Content ◽  
Breaking Dormancy ◽  
Seed Extracts

Siberian C peach (Prunus persica L.) seeds were stratified at 5 and 20C. DWs and soluble protein content remained constant regardless of stratification temperature and duration. Seed extracts subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a decrease in the intensity of nine polypeptides in the cotyledons of seeds held at 5C during weeks 5 through 8, coinciding with an increase in germination capacity. These changes were confined to cotyledons held at 5C, and were observed only when the seeds were able to germinate. The effects of stratification and the imbibition degree on changes in the protein content of seeds of two additional peach biotypes (`Farouki' and `Maloussi') were also evaluated. Germination of fully imbibed seeds at 20C increased steadily as stratification time at 5C increased. Partially imbibed seeds (25 % or 50% of full imbibition) did not germinate regardless of stratification time. However, when these seeds were soaked in water after stratification, their germination paralleled that of fully imbibed seeds. Thus, dormancy was broken, even though the seeds could not germinate. Changes in protein profiles in fully imbibed seeds confirmed those previously reported for Siberian C seeds. Similar changes occurred in cotyledons of partially imbibed seeds during stratification at 5C, but at a slower rate. Those changes were, however, delayed by partial imbibition, whereas germination capacity (ability to germinate when fully imbibed) was not. Changes in cotyledon protein profiles were not affected by removing the embryonic axis before stratification, a result indicating that such changes are not controlled by the axis. Gibberellic acid (GA3 induced 35 % to 40% germination of nonchilled seeds. It hastened the loss of protein band intensity in `Farouki' but not in `Maloussi'. However, GA3-treated seeds germinated before any visible changes occurred in protein profiles. We conclude that the effects of chilling on breaking dormancy are independent of its effects on the protein changes observed in this study.

Differences in structure, allergenic protein content and pectate lyase enzyme activity of some Cupressaceae pollen

2018 ◽  
Vol 43 (4) ◽  
pp. 435-446
Author(s):  
Aydan Acar Şahin ◽  
Belma Aslım ◽  
Sema Tan ◽  
Şenol Alan ◽  
Nur Münevver Pınar
Keyword(s):  
Protein Content ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Pectate Lyase ◽  
Polyacrylamide Gel ◽  
Pollen Allergen ◽  
Allergenic Protein ◽  
Diagnosis And Therapy ◽  
Cupressus Arizonica

Abstract Objective Cupressaceae pollen has commonly been reported to be an important aeroallergen and causal factor of spring, autumn and winter pollinosis in many countries. The aim of this study was to compare of the structure and allergenic protein content of Cupressus arizonica Greene., Cupressus sempervirens L. and Juniperus oxycedrus L. pollen in detail and contribute to Cupressaceae pollen allergen diagnosis and therapy studies in Turkey. Methods The pollen structure were examined by LM and SEM. Pollen protein content was investigated by Bradford protein assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot analysis and two-dimensional polyacrylamide gel electrophoresis (2DE PAGE), respectively. Pectate lyase (PL) enzyme activities were compared. Immunoblotting was carried out by using extracts of the three taxa pollen collected from Turkey. Results All three taxa was found very similar in terms of pollen morphology however, intine thickness was prominently different. Cupressus arizonica pollen extracts showed the lowest PL activity. Five sera specific IgE of all allergic subjects showed reaction with only C. arizonica pollen extracts. Conclusions As a conclusion, the pollen structure, protein function or protein structure and isoforms of allergens could affects allergenic properties of the pollen. This study also may help to improve the Cupressaceae pollen allergen diagnosis and therapy.

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