Identification and Structure Prediction of Human Septin-4 as a Biomarker for Diagnosis of Asthenozoospermic Infertile Patients—Critical Finding Toward Personalized Medicine

Semen parameters are been found as a key factor to evaluate the count and morphology in the given semen sample. The deep knowledge of male infertility will unravel with semen parameters correlated with molecular and biochemical parameters. The current research study is to identify the motility associated protein and its structure through the in-silico approach. Semen samples were collected and initial analysis including semen parameters was analyzed by using the World Health Organization protocol. Semen biochemical parameters, namely, seminal plasma protein concentration, fructose content, and glucosidase content were calculated and evaluated for correlation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) were carried out for identification of Septin-4 presence in the semen sample. Mascot search was done for protein conformation and in-silico characterization of Septin-4 by structural modeling in Iterative Threading Assembly Refinement (I-TASSER). Twenty-five nanoseconds molecular dynamics (MD) simulations results showed the stable nature of Septin-4 in the dynamic system. Overall, our results showed the presence of motility-associated protein in normospermia and control samples and not in the case of asthenospermia and oligoasthenospermia. Molecular techniques characterized the presence of Septin-4 and as a novel biomarker for infertility diagnosis.

Download Full-text

Surface coat proteins of the pine wood nematode, Bursaphelenchus xylophilus: profiles of stage- and isolate-specific characters

Nematology ◽

10.1163/156854109x447006 ◽

2009 ◽

Vol 11 (3) ◽

pp. 429-438 ◽

Cited By ~ 9

Author(s):

Yuko Takeuchi ◽

Ryoji Shinya ◽

Kouichi Kuroda ◽

Natsuko Miura ◽

Kazuyoshi Futai ◽

...

Keyword(s):

Life Stage ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Bursaphelenchus Xylophilus ◽

Molecular Techniques ◽

Surface Coat ◽

Coat Proteins ◽

Pine Wood Nematode ◽

Protein Patterns ◽

Pine Wood

AbstractThe present study was made to determine the binding patterns of several lectins to the surface coat (SC) proteins of various isolates and developmental stages of the pine wood nematode (PWN), Bursaphelenchus xylophilus. Also, the detailed characteristics of the SC proteins were profiled by using molecular techniques. The lectin-binding study demonstrated the stage-specific characters of SC in binding to the lectin, wheat germ agglutinin (WGA). WGA binding was observed only to the outer surfaces of third-stage propagative juveniles and to the egg shells, and this occurred more frequently in virulent than in avirulent PWN isolates. A greater variety of lectins bound to eggs than to any other life stage. For characterisation, the SC proteins extracted were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and analysed by lectin blotting. The results showed that the carbohydrate and protein patterns of the SC of the PWN changed during nematode development.

Download Full-text

Use of Seed Protein Profiles to Characterize Peanut Cultivars1

Peanut Science ◽

10.3146/i0095-3679-21-2-18 ◽

1994 ◽

Vol 21 (2) ◽

pp. 152-159 ◽

Cited By ~ 6

Author(s):

C. M. Bianchi-Hall ◽

R. D. Keys ◽

H. T. Stalker

Keyword(s):

Seed Protein ◽

Cultivar Identification ◽

Storage Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Genetic Material ◽

Seed Storage Proteins ◽

Molecular Techniques ◽

Protein Profiles ◽

Sds Page

Abstract In the last 10 to 15 yr, the development of biotechnology and molecular techniques has allowed great advancements toward the identification of cultivars among plant species. In legumes, the success of cultivar identification depends on the species under investigation, the type and variability of genetic material found in cultivars, and the technology used for investigations. In this study, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to assess diversity of peanut (Arachis hypogaea L.) seed protein profiles. The objectives of this investigation were a) to assess diversity of protein profiles in peanuts for cultivar identification using SDS-PAGE and b) to determine the extent of variability of seed storage proteins (SSP) among samples of cultivars originating from different locations. The first study included 34 cultivars grown at Lewiston, NC and the second one included nine cultivars grown at six locations. The results of both studies indicated that it is possible to differentiate between subspecies but not to associate a particular profile with only one specific cultivar. Within subspecies, cultivars clustered in more than one group and most cultivars that grouped together were genetically related.

Download Full-text

Protein Profiles from Intact Cells as a Tool in Bifidobacterium Characteristics

Polish Journal of Microbiology ◽

10.33073/pjm-2012-041 ◽

2012 ◽

Vol 61 (4) ◽

pp. 305-310 ◽

Cited By ~ 1

Author(s):

ADAM WAŚKO ◽

MAGDALENA POLAK-BERECKA ◽

MICHAŁ KALITA

Keyword(s):

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Molecular Techniques ◽

Group Method ◽

Intact Cells ◽

Arithmetic Average ◽

Pair Group ◽

Sds Page ◽

Liquid Chromatography Tandem Mass ◽

Electrophoresis Separation

In this study sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) profiles were analysed and differences were confirmed by a unweighted pair group method with arithmetic average (UPGMA) analysis between bifidobacterial species, such as B. infanis ATCC1567, B. bifidum Bb-12, B. longum KN29, B. catenulatum KD14, and B. animalis BI30. Two dimensional electrophoresis separation profiles were compared, and the most characteristic spots were characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We propose proteins extracted from intact cells as an additional trait for bifidobacteria characterization, together with molecular techniques, which can be used to analyze bacterial protein polymorphism and to distinguish among species.

Download Full-text

Plasma protein concentrations of the young and adult Amazona brasiliensis parrots

Veterinarski glasnik ◽

10.2298/vetgl170117008s ◽

2017 ◽

Vol 71 (1) ◽

pp. 44-51

Author(s):

Elizabeth Schmidt ◽

Patricia Serafini ◽

Elenise Sipinski ◽

Antonio Paulillo

Keyword(s):

Plasma Protein ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Clinical Pathology ◽

Specific Protein ◽

Major Protein ◽

Plasma Protein Concentration ◽

Protein Gel ◽

Sds Page

Introduction. The Red-tailed Amazon parrot (Amazona brasiliensis) is an endangered species of the Psittacine family, and for which various data are important for a comprehensive preservation plan. Data about plasma protein gel electrophoresis of Amazon parrot blood are scarce. The purpose of this study was to determine plasma protein concentrations and concentrations of major protein bands in blood of young and adult Red-tailed Amazon parrot (Amazona brasiliensis). Materials and Methods. Blood samples from eight young and eight adult healthy free-living parrots were obtained. Plasma protein concentration and fractions were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Mann-Whitney U test was used to compare variables. Results and Conclusions. Six major protein bands with the following molecular weights were identified by SDS-PAGE: 170 kDa, 117 kDa, 85 kDa (putative ovotransferrin), 60 kDa, 45 kDa and 23 kDa. Adult parrots had significantly higher concentrations of total proteins, albumin and other proteins with similar mobility (around 60 kDa). Young birds had significantly higher levels of 23kDa proteins. The concentration of putative ovotransferrin (85 kDa) was not different between young and adult parrots. Plasma protein gel electrophoresis patterns in Red-tailed Amazon parrots are similar between young and adult animals, but specific protein bands differ in their absolute concentrations. This finding should be taken into consideration when clinical pathology data are analysed.

Download Full-text

Liver proteome alterations in psychologically distressed rats and a nootropic drug

PeerJ ◽

10.7717/peerj.11483 ◽

2021 ◽

Vol 9 ◽

pp. e11483

Author(s):

Raquel González-Fernández ◽

Mariana Grigoruţă ◽

Sarahi Chávez-Martínez ◽

Eliel Ruiz-May ◽

José Miguel Elizalde-Contreras ◽

...

Keyword(s):

Liver Disease ◽

Psychological Stress ◽

Biochemical Parameters ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

The Body ◽

Alcoholic Fatty Liver ◽

Proteomic Approach ◽

Wide Range ◽

Liver Proteome

Background Chronic psychological distress is considered today a pandemic due to the modern lifestyle and has been associated with various neurodegenerative, autoimmune, or systemic inflammation-related diseases. Stress is closely related to liver disease exacerbation through the high activity of the endocrine and autonomic nervous systems, and the connection between the development of these pathologies and the physiological effects induced by oxidative stress is not yet completely understood. The use of nootropics, as the cognitive enhancer and antioxidant piracetam, is attractive to repair the oxidative damage. A proteomic approach provides the possibility to obtain an in-depth comprehension of the affected cellular processes and the possible consequences for the body. Therefore, we considered to describe the effect of distress and piracetam on the liver proteome. Methods We used a murine model of psychological stress by predatory odor as a distress paradigm. Female Sprague-Dawley rats were distributed into four experimental groups (n = 6 − 7/group) and were exposed or not to the stressor for five days and treated or not with piracetam (600 mg/kg) for six days. We evaluated the liver proteome by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D-SDS-PAGE) followed by liquid chromatography-tandem mass spectrometry (GeLC-MS/MS). Besides, we analyzed the activity of liver antioxidant enzymes, the biochemical parameters in plasma and rat behavior. Results Our results showed that distress altered a wide range of proteins involved in amino acids metabolism, glucose, and fatty acid mobilization and degradation on the way to produce energy, protein folding, trafficking and degradation, redox metabolism, and its implications in the development of the non-alcoholic fatty liver disease (NAFLD). Piracetam reverted the changes in metabolism caused by distress exposure, and, under physiological conditions, it increased catabolism rate directed towards energy production. These results confirm the possible relationship between chronic psychological stress and the progression of NAFLD, as well as we newly evidenced the controversial beneficial effects of piracetam. Finally, we propose new distress biomarkers in the liver as the protein DJ-1 (PARK7), glutathione peroxidase 1 (GPX), peroxiredoxin-5 (PRDX5), glutaredoxin 5 (GLRX5), and thioredoxin reductase 1 (TXNDR1), and in plasma as biochemical parameters related to kidney function such as urea and blood urea nitrogen (BUN) levels.

Download Full-text

Relationship between asthenozoospermia and selected macroscopic, microscopic and biochemical semen parameters

Diagnostyka Laboratoryjna ◽

10.5604/01.3001.0013.7965 ◽

2017 ◽

Vol 53 (2) ◽

pp. 71-78

Author(s):

Anna Świerczyńska-Ciepłucha ◽

Katarzyna Marchlewska ◽

Renata Walczak-Jędrzejowska ◽

Eliza Filipiak ◽

Jolanta Słowikowska-Hilczer

Keyword(s):

Citric Acid ◽

Biochemical Parameters ◽

Sperm Count ◽

World Health ◽

Semen Parameters ◽

Lower Percentage ◽

Sperm Number ◽

Normal Morphology ◽

Sperm Analysis ◽

Activity Marker

Asthenozoospermia is a sperm motility disorder in which <32% of spermatozoa show progressive motility, according to the World Health Organization definition (WHO, 2010). Among causes of male infertility asthenozoospermia accounts for nearly 19%. The aim of the study was to determine the relationship between asthenozoospermia and selected macroscopic, microscopic and biochemical parameters of semen. The semen of 112 males from infertile couples was studied. Basic sperm analysis was performed by manual method according to the WHO 2010 guidelines. Macroscopic parameters (volume, pH) and microscopic (total sperm number and concentration, percentage of vital spermatozoa and with normal morphology) were evaluated. Biochemical parameters (neutral α-glucosidase activity – epididymis activity marker, fructose concentration – seminal vesicles activity marker, and citric acid – prostatic activity marker) were evaluated by spectrophotometric method. Patients with asthenozoospermia had a statistically significant decrease in the ejaculate volume, lower total sperm number and lower percentage of vital sperms, as well as lower fructose and citric acid concentrations in comparison to those without asthenozoospermia. There were statistically significant positive correlations between the percentage of spermatozoa showing progressive movement and the ejaculate volume, total number and concentration of spermatozoa, percentage of with normal vitality and morphology, as well as total fructose concentration in the ejaculate. Conclusions: Asthenozoospermia may be associated with abnormal macroscopic and microscopic semen parameters such as reduced ejaculate volume, reduced sperm count, reduced spermatozoa and normal morphology, and reduced biochemical parameters. Co-occurrence of abnormal macroscopic, microscopic and biochemical parameters of semen may indicate a common etiological factor for these disorders.

Download Full-text

Cytogenetic and molecular characterization of a durum alien disomic addition line with enhanced tolerance to Fusarium head blight

Genome ◽

10.1139/g09-014 ◽

2009 ◽

Vol 52 (5) ◽

pp. 467-483 ◽

Cited By ~ 49

Author(s):

Prem P. Jauhar ◽

Terrance S. Peterson ◽

Steven S. Xu

Keyword(s):

Fusarium Head Blight ◽

Chromosome Pair ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Molecular Techniques ◽

Intergeneric Hybridization ◽

Specific Marker ◽

Addition Line ◽

Head Blight ◽

Fhb Resistance

Current durum wheat ( Triticum turgidum L. subsp. durum (Desf.)) cultivars have little or no resistance to Fusarium head blight (FHB), a ravaging disease of cereal crops. A diploid wheatgrass, Lophopyrum elongatum (Host) Á. Löve (2n = 2x = 14, EE genome), is an excellent source of FHB resistance. Through an extensive intergeneric hybridization using durum cultivar Langdon, we have developed a disomic alien addition line, named DGE-1 (2n = 28 + 2), with a wheatgrass chromosome pair. We used a unique method for isolating the addition line taking advantage of unreduced gametes functioning in Langdon × L. elongatum F1 hybrids in their first backcross to the Langdon parent, resulting in 35-chromosome plants from which we derived DGE-1. The addition line DGE-1 has a plant type similar to its Langdon parent, although it is shorter in height with narrower leaves and shorter spikes. It is meiotically and reproductively stable, generally forming 15 bivalents with two chiasmata each. The alien chromosome pair from the grass confers FHB resistance to the addition line, which has less than 21% infection on the visual scale, mean = 6.5%. Using various biochemical and molecular techniques (Giemsa C-banding, fluorescent genomic in situ hybridization (fl-GISH), chromosome-specific simple sequence repeat (SSR) markers, targeted region amplified polymorphism (TRAP) markers, and sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE)), we have shown that the extra chromosome involved is 1E of L. elongatum. This is the first time that FHB resistance has been discovered on chromosome 1E. We have established a chromosome-specific marker for 1E that may be used to screen fertile hybrid derivatives and durum addition lines for this chromosome that confers FHB resistance.

Download Full-text

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646313 ◽

1992 ◽

Vol 68 (05) ◽

pp. 534-538 ◽

Cited By ~ 5

Author(s):

Nobuhiko Yoshida ◽

Shingi Imaoka ◽

Hajime Hirata ◽

Michio Matsuda ◽

Shinji Asakura

Keyword(s):

Molecular Weight ◽

Sodium Dodecyl Sulfate ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Tetraacetic Acid ◽

Fibrin Monomer ◽

Sds Page ◽

Thrombotic Tendency ◽

Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

Download Full-text

The Recovery of Factor VIII from Fresh-Frozen, Indated and Outdated Human Plasma

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648010 ◽

1976 ◽

Vol 36 (01) ◽

pp. 071-077 ◽

Cited By ~ 2

Author(s):

Daniel E. Whitman ◽

Mary Ellen Switzer ◽

Patrick A. McKee

Keyword(s):

Factor Viii ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

The United States ◽

Fresh Frozen Plasma ◽

Red Cross ◽

Random Samples ◽

Fresh Frozen ◽

Factor Viii Concentrates ◽

Frozen Plasma

SummaryThe availability of factor VIII concentrates is frequently a limitation in the management of classical hemophilia. Such concentrates are prepared from fresh or fresh-frozen plasma. A significant volume of plasma in the United States becomes “indated”, i. e., in contact with red blood cells for 24 hours at 4°, and is therefore not used to prepare factor VIII concentrates. To evaluate this possible resource, partially purified factor VIII was prepared from random samples of fresh-frozen, indated and outdated plasma. The yield of factor VIII protein and procoagulant activity from indated plasma was about the same as that from fresh-frozen plasma. The yield from outdated plasma was substantially less. After further purification, factor VIII from the three sources gave a single subunit band when reduced and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These results indicate that the approximately 287,000 liters of indated plasma processed annually by the American National Red Cross (ANRC) could be used to prepare factor VIII concentrates of good quality. This resource alone could quadruple the supply of factor VIII available for therapy.

Download Full-text

Low Molecular Weight Trypsin-Plasmin Inhibitors Isolated from Papain Treated Urinary Trypsin Inhibitor

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657115 ◽

1982 ◽

Vol 47 (01) ◽

pp. 014-018 ◽

Cited By ~ 13

Author(s):

H Sumi ◽

N Toki ◽

S Takasugi ◽

S Maehara ◽

M Maruyama ◽

...

Keyword(s):

Trypsin Inhibitor ◽

Gel Electrophoresis ◽

Specific Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Chromatography ◽

Urinary Trypsin Inhibitor ◽

Plasmin Activity ◽

Molecular Weights ◽

Plasmin Inhibitors

SummaryPapain treatment of human urinary trypsin inhibitor (UTI67; mol. wt. 43,000 by SDS-polyacrylamide gel electrophoresis, specific activity 1,897 U/mg protein) produced four new protease inhibitors, which were highly purified by gel chromatography on Sephadex G-100 and isoelectric focusing. The purified inhibitors (UTI26, UTI9-I, UTI9-II, and UTI9-III) were shown to be homogeneous by polyacrylamide disc gel electrophoresis, and had apparent molecular weights of 26,000, 9,000, 9,000, and 9,800, respectively, by sodium dodecyl sulfate gel electrophoresis. During enzymatic degradation of UTI67, the amino acid compositions changed to more basic, and the isoelectric point increased from pH 2.0 (UTI67) to pHs 4.4, 5.2, 6.6, and 8.3 (UTI26, UTI9-I, UTI9-II, and UTI9-III), respectively. Both the parent and degraded inhibitors had anti-plasmin activity as well as antitrypsin and anti-chymotrypsin activities. Much higher anti-plasmin/anti-trypsin and anti-plasmin/anti-chymotrypsin activities were observed in the degraded inhibitors than in the parent UTI67. They competitively inhibited human plasmin with Ki values of 1.13 X 10-7 - 2.12 X 10-6 M (H-D-Val-Leu-Lys-pNA substrate). The reactions were very fast and the active site of the inhibitors to plasmin was thought to be different from that to trypsin or chymotrypsin.

Download Full-text