Gender identification of fish species is carried out mainly by examining external morphological characteristics, which in general, it is very complex and not always a reliable approach. Electrophoresis of plasma proteins can be used as an alternative and useful molecular tool for a more precise sex determination. The presence of female specific proteins in the plasma is a starting point for the application of this technique. In this study, reducing discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was applied to analyze plasma proteins of male and female koi carp (Cyprinus carpio haematopterus). Image analyses of electrophoregrams with resolved plasma proteins by SDS-PAGE showed that it is an appropriate technique to discriminate male from female samples. It is based on the presence of apolipoprotein B-100 which can be used as a suitable marker. Further amino acid characterization of apolipoprotein B-100 confirmed that it is a specific protein for female individuals.
: Diagnosing Helminthiasis through Coprological Examination. Janssen Research Foundation , Beerse, Belgium, 34 – 36 T hompson , J.D., H iggins , D.G., G ibson , T.J. (1994). CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res ., 22: 4673 – 4680 T oledo , R., B ernal , M.D., M arcilla , A. (2011). Proteomics of foodborne trematodes, J. Proteomics, 74: 1485 – 1503. DOI: 10.1016/j. jprot.2011.03.029 U padhyay , A.K., K umar , M. (2002). SDS-PAGE
The differences in protein and enzymatic profiles of infective larvae (L3) of Trichostrongylus colubriformis, both induced and non-induced to hypobiosis, have been evaluated by means of SDS-PAGE and densitometric analysis as well as by semiquantitative micromethod API-ZYM (Bio-Mérieux, France). Quantitative differences were identified in protein levels between the induced and non-induced larvae, where the amount of two polypeptides (200–220 kDa) decreased in range 32.3–35.4 % and the amount of six polypeptides (20–28 kDa) increased in range 20.0–27.0 % in the samples of induced larvae. In contrast to non-induced larvae, on gelatin-substrate gel in L3
in vitro released (IVR) proteases from larvae conditioned to hypobiosis, zones of proteolysis were observed between 21 and 34 kDa.
Background: Vector control is a key strategy for eradication of filariasis, but it is limited, possibly due to rapid propagation from global warming. In Thailand, Mansonia mosquitoes are major vectors of filariasis caused by Brugia malayi filarial nematodes. However, little is yet known about vector biology and host-parasite relationship.
Objectives: Demonstrate the preliminary data of salivary gland morphology and protein profile of human filarial mosquitoes M. uniformis.
Methods: Morphology of M. uniformis salivary gland in both sexes was comparatively studied under a light microscope. Total protein quantization and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to compare protein profile between male and female. In addition, quantitative analysis prior to and after blood feeding was made at different times (0, 12, 24, 36, 48, 60, and 72 hours).
Results: Total salivary gland protein of males and females was 0.32±0.03 and 1.38±0.02 μg/pair gland, respectively. SDS-PAGE analysis of the female salivary gland protein prior to blood meal demonstrated twelve bands of major proteins at 21, 22, 24, 26, 37, 39, 44, 53, 55, 61, 72, and 100 kDa. Compared to female, male salivary gland was composed of seven major protein bands at 39, 44, 53, 55, 61, 83, and 100 kDa. Quantitative study after blood feeding revealed that protein of 37 kDa decreased gradually whereas proteins of 61 and 83 kDa started to increase dramatically at 24 hours. It was postulated that the 37 kDa band, found only in the female, might serve as a candidate molecule for facilitating blood feeding.
Conclusion: Morphology and protein components of M. uniformis salivary glands might relate to blood feeding process and filarial disease transmission.
N′-Nitrosodiethylamine (NDEA) is an effective hepatotoxicant, carcinogen and mutagen. NDEA-induced hepatic necrosis, through metabolic activation by CYP2E1, is an extensively used experimental model. In the present study, we analysed the dose- and time-dependent effect of NDEA on hepatic damage, RBC rheology and proteomic profile in male Wistar rats. The rats, 5–6 weeks old, were divided into four groups: Group-1 served as control and received normal saline, Group-2 received a single dose of 200 mg/kg body weight NDEA intraperitoneally (i.p.) and the animals were sacrificed after one week; the rats of Group-3 received a single dose of 100 mg/kg body weight NDEA and were sacrificed after one week; Group-4 received 100 mg/kg body weight/wk NDEA for two weeks and were then sacrificed. Various biochemical parameters such as ALT, AST, ALP and bilirubin were determined. Further, RBC rheology, histopathology (H&E staining) of liver biopsies and polypeptide profiling (SDS-PAGE) in sera and liver sections were also carried out both in control and NDEA treated groups. Our results showed a significant increase in all the biochemical parameters of the liver function test (p<0.05). In NDEA treated categories dacryocytes (tear drop cells), schistocytes (fragmented cells), codocytes (target cells), acanthocytes (spur cells) and ovalocytes (oval cells) were observed. H & E stained liver biopsies treated with NDEA showed abnormal liver architecture with severe haemorrhage, neutrophilic infiltration and dysplastic hepatocytes manifested in a dose-dependent manner. Software analysis of SDS-PAGE of control and NDEA treated rat sera and liver revealed qualitative and quantitative differences in polypeptide composition. Based on the presence/absence, polypeptides were classified in three different categories: (1) house-keeping, present in all the groups investigated; (2) novel, present in either control or NDEA treated group at any given time; (3) differential expression, showing quantitative differences. Our study indicates a dose and time-dependent hepatocellular damage and proteome profile which is likely due to NDEA-mediated oxidative stress in rats.
Echinacea purpurea (L.) Moench, a member of the Asteraceae family, is a plant rich in flavonoids, essential oils, phenolic compounds, saponins, polysaccharides and glycoproteins. The aim of the study was to evaluate the protein content in dried roots of Echinacea purpurea (L.) Moench after homogenization of roots with liquid nitrogen, extraction in 0.01 mol L-1 phosphate-buffered saline (PBS) and purification followed by fractionation of proteins using gel filtration chromatography. Total concentration of proteins was measured using the Bradford method, and evaluation of the molecular mass of proteins was accomplished by applying the SDS-PAGE gel electrophoresis. The Bradford assay revealed that the highest concentration of proteins in fractions collected after gel filtration chomatography was 4.66–6.07 mg mL-1. Glycoproteins, alkamides and polysaccharides in roots of Echinacea purpurea (L.) Moench are chemical compounds that are responsible for their immunomodulatory properties. However, information about the difference of protein contents in fresh and dried roots of E. purpurea is insufficient.
Maize (Zea mays L.) constitutes one of the most important crops worldwide with multi-billion dollar annual revenue. The plant is however a good substrate for growth, development and activity of filamentous fungi. A large number of fungal species causes spoilage and accumulation of mycotoxins. Plants restrict the hyphal growth by producing pathogenesis related proteins. So far 17 groups of such proteins are identified. PR-5 group comprises of the thaumatin-like proteins (TLPs), which have diverse modes of actions and act at various stages of fungal attack. Zeamatin-like protein (ZLP) is a member of TLPs, which is basically localized in seeds with enhanced expression during physiological growth and cellular differentiation. However a basal quantity is found in the leaves of many crop plants. Here we report the response of maize plant tissues against A. niger inoculation by measuring the variation in expression profile of a zeamatin-like gene. Conventional PCR coupled with RT-qPCR identifies a significant change in the expression magnitude of ZLP in pre- and post-inoculated plant samples. SDS-PAGE, followed by antimicrobial activities against A. niger, E.coli, P. aeruginosa, B. cereus, S. aureus and S. typhimurium, however, do not register a direct relationship with enhancement in gene expression. It is in line with the fact that response to pathogenesis in plants is a multigenic activity involving a series of responsible/induced genes. The assay developed is useful in primary sorting out of the maize hybrids with respect to their resistance against Aspergillus spp., especially in areas with high rate of incidence of fungal pathogenesis.
Allergens from Fusarium solani Identified by Immunoblotting in Asthma Patients In Iran
We extracted Fusarium solani antigens to evaluate specific anti-F. solani IgE in fifty-one patients with asthma (33 men and 18 women) and in 22 non-atopic healthy subjects (15 men and 7 women). F. solani strains were cultured in Sabouraud glucose agar and subjected to cell disruption using the freeze-and-thaw method. The obtained cytoplasmic extracts were analysed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Sensitisation to F. solani antigens has been evaluated in asthmatic patients using the immunoblotting assay. The SDS-PAGE identified 29 protein bands in the cytoplasmic extracts of F. solani isolates, with molecular weights ranging from 24 kDa to 112 kDa. Immunoblotting detected specific anti-F. solani IgE antibody in all asthma patients, but not in the control group. The predominant reactive allergens in patients corresponded to the bands with molecular weights of 24 kDa, 58.5 kDa, 64.5 kDa, 69 kDa, 72 kDa, and 97 kDa. Our results suggest that various allergenic components of F. solani may produce symptoms of asthma in susceptible individuals and they call for further research.
urinary β2-microglobulin and NAG to creatinine vary with age in children. Pediatr Int. 2015; 57: 79-84. 13. Tsukahara H, Hiraoka M, Kuriyama M, Saito M, Morikawa K, Kuroda M, Tominaga T, Sudo M. Urinary alpha 1-microglobulin as an index of proximal tubular function in early infancy. Pediatr Nephrol. 1993; 7: 199-201. 14. Grillenberger A, Weninger M, Lubec G. Determination of urinary low molecular weight proteins for the diagnosis of tubular damage. Padiatr Padol. 1987; 22: 229-34. 15. Lau YK, Woo KT. SDS-PAGE is underutilized as a tool for investigating renal patients
Flower development and senescence was studied in Narcissus tazetta ‘Kashmir Local’. Flower development was divided into six stages (I-VI), from the tight bud stage to the senescent stage. Flower fresh and dry weight increased as the flowers progressed from bud to bloom and then declined during senescence. Membrane permeability of tepal tissues increased as the flower progressed through various stages. The content of sugars (reducing and total) increased during flower development and declined thereafter during senescence. The α-amino acid content registered an increase during flower development with a concomitant decrease in soluble protein content. The SDS-PAGE of protein extracts from tepal tissues revealed a general decrease in the expression of some high molecular weight proteins and an increase in low molecular weight proteins during flower development and senescence. It may be suggested that the flower senescence of N. tazetta may be linked to the protein turnover and sugar status of the perianth tissues and that these newly synthesized proteins may be involved in proteolysis. At this stage, it is not known whether these polypeptides play an important role in senescence but revealing the nature of these proteins can provide new insights into the pathways executing senescence in this flower system.