scholarly journals Characterization of Streptococcus sanguis molecular receptors for Streptococcus mutans binding molecules

2016 ◽  
Vol 49 (4) ◽  
pp. 213
Author(s):  
Deby Kania Tri Putri ◽  
Indah Listiana Kriswandini ◽  
Muhammad Luthfi
Keyword(s):  
Molecular Weight ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Profile Analysis ◽  
Protein Profile ◽  
Tooth Surface ◽  
Molecular Receptors ◽  
Streptococcus Sanguis ◽  
Sds Page

Background: Dental caries is a major problem in oral cavity. If dental caries causes cavity, the structure of dental hard tissue will not be reversible because of damage in the structure of the hard tissue. The early pathogenesis mechanism of dental caries is an adhesion interaction between cariogenic Streptococcus mutans microorganisms and tooth surface pellicles. The attachment involves a specific molecular component interaction between the bacterial complement molecules and the surface of the host. Streptococcus sanguis as a dominant ecology at the beginning of bacterial plaque aggregation will colonize the tooth surface earlier than S. mutans. The surface of bacterial cells can express some adesin. The bacteria also can express receptors for adhesins of other bacteria. Specific receptors for adhesions of S. Mutans bacteria are not only found in the pellicles, but also present in pioneer bacteria, such as S. sanguis. Adhesion between those bacteria is called as coagregation. Purpose: This study aimed to analyze the characterization of Streptococcus sanguis molecular receptors for Streptococcus mutans binding molecules. Method: This study used a sonication method for protein isolation of S. mutans and S. sanguis bacterial biofilms, as well as electrophoresis method using 12 % SDS-PAGE gel and Western Blot analysis. Result: Results of the protein profile analysis of S. mutans biofilms using 12% SDS-PAGE showed that there were 17 bands, each of which molecular weights was 212, 140, 81, 65, 61, 48, 45, 44, 40, 39, 33 , 25, 23, 19, 17, 12, and 11 kDa. On the other hand, results of the protein profile analysis of S. sanguis biofilms using 12% SDS-PAGE showed that there were 15 bands, each of which molecular weight was 130, 85, 65, 61, 48, 46, 40, 37, 29, 25, 23, 21, 17, 15, and 12 kDa. And, results of the analysis of S. sanguis receptor molecules using Western blot showed that there were three bands, each of which molecular weight was 130, 85, and 40 kDa. Conclusion: S. sanguis bacteria have specific receptor molecules for S. mutans bacteria with the molecular weight of 130, 85, and 40 kDa.

scholarly journals Cloning, Expression, Purification, and Preliminary Characterization of Single-Crystal X-Ray Diffraction of Glucosyltransferase B of Streptococcus mutans

2019 ◽  
Vol 14 (5) ◽  
pp. 1934578X1984933
Author(s):  
Joshua L. Mieher ◽  
Norbert Schormann ◽  
Manisha Patel ◽  
Hui Wu ◽  
Champion Deivanayagam
Keyword(s):  
Dental Caries ◽  
Oral Cavity ◽  
Single Crystal ◽  
Streptococcus Mutans ◽  
Tooth Enamel ◽  
Tooth Surface ◽  
X Ray Diffraction ◽  
Persistent Disease ◽  
X Ray

Dental caries characterized by acid damage of tooth enamel is a persistent disease that begins with the formation of biofilms on the tooth surface. The secreted glucosyltransferases enable Streptococcus mutans to synthesize extracellular glucan polymers using ingested starch within the oral cavity, which eventually results in the production of acid, a contributing factor to cariogenesis. In this paper, we report the cloning, expression, purification, crystallization, and preliminary X-ray diffraction characterization of glucosyltransferase B.

scholarly journals Morphology, Protein Profile and the Level of Infestation of Bedbugs (Hempitera: Cimicidae) in the Halls of Residence of a Higher Institution in Lagos.

2019 ◽  
Vol 1 ◽  
pp. 96-104
Author(s):  
K A Kemabonta ◽  
M O Ajiboye
Keyword(s):  
Staining Method ◽  
Profile Analysis ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Morphological Characterization ◽  
Sds Page ◽  
Higher Institution ◽  
Cimex Hemipterus ◽  
Very High

Morphology, protein profile and the level of Bedbug infestations were carried out at a University in Lagos, Nigeria after an outcry of high infestation by the students in halls of residence. Three bedbugswere each collected from nine halls of residence for morphological characterization of adults and to carry out protein profile analysis usingSodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Coomassie staining method. Level of infestation of Bedbugs was carried out in two female halls A and Busing blood smeared on the walls. All bedbugs found were Cimex hemipterus. 29.8% of the rooms in Hall Bhad no bedbug infestation, 23.9%, 20.2%, 15.4% and 10.6% had low, average, high and very high levels of bedbug infestation respectively while in A hall, it was 21.4%, 30.5%, 24.4%, 14.5% and 9.2% respectively. In A hall, 4.6%, 16.8% and 0.8% had one, two and three out of the four mattresses in the room infested with bedbugs respectively while 56.5% had all the four mattresses infested with bedbugs. In B hall, 1.6%, 16.0% and 0% had one, two and three mattresses infested with bedbugs respectively while the remaining 52.7% had all the four mattresses in the room infested with bedbugs. There was a significant relationship between the level of infestation and the number of mattresses infested in each hall (P < 0.01).The protein profile analysis of bedbugs did not show the protein bands clearly because of the low soluble protein content of Cimex hemipterus and the detection limit of Coomassie stain.

scholarly journals Biological and Immunogenicity Property of IgY Anti S. mutans ComD

2016 ◽  
Vol 10 (1) ◽  
pp. 308-314 ◽  
Author(s):  
E.W. Bachtiar ◽  
B.M. Bachtiar ◽  
R.D. Soejoedono ◽  
I.W. Wibawan ◽  
A. Afdhal
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Dna Vaccine ◽  
Biofilm Formation ◽  
Protein Profile ◽  
Egg Yolk ◽  
Biological Properties ◽  
Coding Region ◽  
Dna Coding ◽  
Sds Page

Objective:This study aims to elucidate the effect of IgY anti ComD on the biological properties ofStreptococcus mutans. (S. mutans)ComD is an interspecies quorum-sensing signaling receptor that plays an important role in biofilm formation byS. mutans.Materials and Methodology:Egg yolk IgY was produced by the immunization of chickens with a DNA vaccine containing the ComD DNA coding region. We evaluated the effect of the antibody on biofilm formation byS. mutansisolated from subjects with or without dental caries. We also assessed the immunoreactivity of the antibody against all isolates, and analyzed the protein profile ofS. mutansby SDS-PAGE.Results:The ComD antibody was successfully induced in the hens’ eggs. It inhibited biofilm formation by allS. mutansisolates. In addition, the expression of some protein bands was affected after exposure to the antibody.Conclusion:IgYanti-S. mutansComD reduces biofilm formation by this bacterium and alters the protein profile ofS. mutans.

scholarly journals Immunohistochemical and immunochemical characterization of a new endothelial cell-specific antigen.

1986 ◽  
Vol 34 (2) ◽  
pp. 209-214 ◽  
Author(s):  
J U Alles ◽  
K Bosslet
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Specific Antigen ◽  
Similar Molecular Weight ◽  
Immunochemical Characterization ◽  
Sds Page

A new monoclonal antibody (moab BW 200) of IgG3 kappa-isotype was generated which recognizes an epitope located on an antigen molecule restricted to human neoplastic and non-neoplastic endothelial cells. The molecular weight of the antigen was determined using immunoprecipitation analysis followed by SDS-PAGE. Despite its similar molecular weight to FVIII-RAG, the antigen detected by moab BW 200 was shown to be different from FVIII-RAG.

scholarly journals Essential Roles of thesppRAFructose-Phosphate Phosphohydrolase Operon in Carbohydrate Metabolism and Virulence Expression byStreptococcus mutans

2018 ◽  
Vol 201 (2) ◽  
Author(s):  
Lin Zeng ◽  
Robert A. Burne
Keyword(s):  
Dental Caries ◽  
Organic Acids ◽  
Streptococcus Mutans ◽  
Biochemical Characterization ◽  
Constitutive Expression ◽  
Extracellular Dna ◽  
Tooth Surface ◽  
Oral Diseases ◽  
Sugar Phosphate ◽  
Content Type

ABSTRACTThe dental caries pathogenStreptococcus mutanscan ferment a variety of sugars to produce organic acids. Exposure ofS. mutansto certain nonmetabolizable carbohydrates, such as xylitol, impairs growth and can cause cell death. Recently, the presence of a sugar-phosphate stress inS. mutanswas demonstrated using a mutant lacking 1-phosphofructokinase (FruK) that accumulates fructose-1-phosphate (F-1-P). Here, we studied an operon inS. mutans,sppRA, which was highly expressed in thefruKmutant. Biochemical characterization of a recombinant SppA protein indicated that it possessed hexose-phosphate phosphohydrolase activity, with preferences for F-1-P and, to a lesser degree, fructose-6-phosphate (F-6-P). SppA activity was stimulated by Mg2+and Mn2+but inhibited by NaF. SppR, a DeoR family regulator, repressed the expression of thesppRAoperon to minimum levels in the absence of the fructose-derived metabolite F-1-P and likely also F-6-P. The accumulation of F-1-P, as a result of growth on fructose, not only inducedsppAexpression, but it significantly altered biofilm maturation through increased cell lysis and enhanced extracellular DNA release. Constitutive expression ofsppA, via a plasmid or by deletingsppR, greatly alleviated fructose-induced stress in afruKmutant, enhanced resistance to xylitol, and reversed the effects of fructose on biofilm formation. Finally, by identifying three additional putative phosphatases that are capable of promoting sugar-phosphate tolerance, we show thatS. mutansis capable of mounting a sugar-phosphate stress response by modulating the levels of certain glycolytic intermediates, functions that are interconnected with the ability of the organism to manifest key virulence behaviors.IMPORTANCEStreptococcus mutansis a major etiologic agent for dental caries, primarily due to its ability to form biofilms on the tooth surface and to convert carbohydrates into organic acids. We have discovered a two-gene operon inS. mutansthat regulates fructose metabolism by controlling the levels of fructose-1-phosphate, a potential signaling compound that affects bacterial behaviors. With fructose becoming increasingly common and abundant in the human diet, we reveal the ways that fructose may alter bacterial development, stress tolerance, and microbial ecology in the oral cavity to promote oral diseases.

scholarly journals Antibiofilm Activity of Mangosteen (Garcinia mangostana L.) Flavonoids against Streptococcus mutans Bacteria

2020 ◽  
Vol 10 (2) ◽  
pp. 48
Author(s):  
Sri Kunarti ◽  
Aulia Ramadhani ◽  
Laskmiari Setyowati
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Tooth Enamel ◽  
Tooth Surface ◽  
Control Group ◽  
Antibiofilm Activity ◽  
Garcinia Mangostana ◽  
Control Group Design ◽  
Biofilm Inhibition ◽  
Significant Difference

Background: Dental caries is one of the most common infectious diseases and often occurs in the community caused by bacteria. Attached bacteria in the tooth surface for a long time will form a biofilm and will lead to demineralization characterized by damage in the structure of the tooth enamel. The bacteria that cause dental caries and can form biofilms is Streptococcus mutans. The bacteria inside biofilms are more resistant to antibacterial agents. Flavonoids in mangosteen pericarp extract can be a cleaner alternative for the anti-biofilm cavity that has properties against Streptococcus mutans. Purpose: To determine the activity of flavonoids in mangosteen pericarp extract at a certain concentration against Streptococcus mutans bacteria. Methods: This study was a laboratory experimental study with a post-test only control group design. Streptococcus mutans were diluted according to the Mc Farland dilution standard 106 in Tryptic Soy Broth (TSB) medium and put in a flexible U-bottom microtiter plate. Then it was incubated for 5x24 hours and checked using crystal violet simple staining to see the formation of biofilms. Flavonoid extract of mangosteen pericarp performed serial dilution in a concentration of 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, 1.56%, and 0.78% was added, and the incubation process were conducted for 1x24 hours. OD (Optical Density) readings were done with a wavelength of 595 nm. Results: There was a significant difference between the test groups and the positive control group. The concentration of 100% had the anti-biofilm activity and showed the value of the highest percentage of inhibition, whilst the concentration of 0.78% showed a minimum biofilm inhibition concentration. The results were demonstrated by a statistical analysis test. Conclusion: Flavonoid extract of mangosteen pericarp at a certain concentration has anti-biofilm activity against Streptococcus mutans biofilm.

Characterization Of The ADP-Induced Fibrinogen Btnd1Ng Sites On Human Platelets Using Photoaffinity Techniques: Some Preliminary Findings

1981 ◽  
Author(s):  
Ellinor I Peerschke ◽  
Mariorie B Zucker ◽  
Avner Rotman
Keyword(s):  
Molecular Weight ◽  
Apparent Molecular Weight ◽  
Human Platelets ◽  
Platelet Membrane ◽  
Membrane Fragment ◽  
Fibrinogen Receptor ◽  
Congenital Deficiency ◽  
Sds Page ◽  
Platelet Membrane Receptor

The interaction of fibrinogen with its, platelet membrane receptor was investigated using 125-labeled fibrinogen which was photoaffinity labeled with a light-sensitive azide. This photoreactive material (125I-NPA-fibr) was indistinguishable from unlabeled fibrinogen as well as from iodinated fibrinogen on SDS-PAGE. It bound specifically to platelets stimulated with ADP and was crosslinked to the platelet membrane after exposure to light ( λ >300 nm) for 4 min. No crosslinking was observed in the presence of EDTA or with platelets that failed to aggregate with ADP either due to the congenital deficiency thrombasthenia or following incubation with EDTA for 8 min at 37° , pH 7.8 and recalcification. SDS-PAGE of platelets bearing crosslinked 125I-NPA-fibr revealed a radiolabeled band of about 450,000 daltons in addition to the 340,000 dalton radioactive band of fibrinogen, suggesting that fibrinogen had been covalently bound to a platelet membrane component with an intact apparent molecular weight of approximately 110,000 daltons. Following reduction, an extra radioactive band was noted at 80,000 daltons. As the A∝-chains of fibrinogen were too weakly labeled to be detected by autoradiography, this indicated that either the Bβ or γchain of fibrinogen was attached to a 25,000-35,000 molecular weight platelet membrane fragment. We conclude that the additional radioactive bands observed after electrophoresis of platelets bearing specifically bound-photoaffinity labeled 125I-fibrinogen most likely represent the binding of the B β or γ chains of fibrinogen to the platelet fibrinogen receptor which may be GPIIb.

Isolation and Partial Characterization of Equine Antithrombin III.

1979 ◽  
Author(s):  
D.W. Estry ◽  
T.G. Bell ◽  
G.H. Tishkoff ◽  
J.C. Mattson ◽  
S.C. Estry
Keyword(s):  
Molecular Weight ◽  
Antithrombin Iii ◽  
Stable Complex ◽  
Sds Page ◽  
At Iii ◽  
Human Thrombin ◽  
Horse Plasma ◽  
Stoichiometric Complex

A protein analogous to human antithrombin III was isolated from fresh horse plasma. The procedure for purification was a modification of Thaler and Schmer’s two-step isolation procedure. The horse protein was homogeneous on 7.5% SDS-PAGE gels and had a molecular weight of 62,000 to 64,000 daltons in both reducing and non-reducing systems (human; 62,300). Rabbit anti-human antithrombin III was used to demonstrate a line of partial identity by Immunoelectrophoresis between the horse and human protein. The horse protein rapidly neutralizes human thrombin (34,000 daltons) and the reaction appears to be greatly potentiated by heparin. In order to establish the formation of 1:1 covalent stoichiometric complex between horse AT III and thrombin (IIa), time studies were run in the presence and absence of heparin. AT III (62,000) at 15 seconds, 2, 5, 10 and 60 minutes formed a stable complex with thrombin (32,000) having a molecular weight of 86,000 daltons. Additional bands developing with time are due to the autolytic capabilities of the uncomplexed IIa. The major autolytic band had a molecular weight of 70,000 daltons. Addition of heparin potentiated the interaction although it did not change the stoichio-metry of the complexes formed. The data accumulated to date demonstrates the similarities between the human and horse protein and the possibilities of using the horse as a model system for the evaluation of AT III replacement therapy in vivo.

scholarly journals Purification and parcial characterization of a hemagglutinating factor (HAF): a possible adhesive factor of the diffuse adherent of Escherichia coli (DAEC)

1996 ◽  
Vol 38 (6) ◽  
pp. 401-406 ◽  
Author(s):  
Yano Tomomasa ◽  
Cleide Ferreira Catani ◽  
Michiko Arita ◽  
Takeshi Honda ◽  
Toshio Miwatani
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Hela Cells ◽  
Immunofluorescence Test ◽  
Native Form ◽  
Bacterial Surface ◽  
E Coli ◽  
Sds Page ◽  
Adhesive Factor

The mannose-resistant hemagglutinating factor (HAF) was extracted and purified from a diffuse adherent Escherichia coli (DAEC) strain belonging to the classic enteropathogenic E. coli (EPEC) serotype (0128). The molecular weight of HAF was estimated to be 18 KDa by SDS-PAGE and 66 KDa by Sephadex G100, suggesting that the native form of HAF consists of 3-4 monomeric HAF. Gold immunolabeling with specific HAF antiserum revealed that the HAF is not a rigid structure like fimbriae on the bacterial surface. The immunofluorescence test using purified HAF on HeLa cells, in addition to the fact that the HAF is distributed among serotypes of EPEC, suggests that HAF is a possible adhesive factor of DAEC strains

scholarly journals Evaluation of biochemical and serological methods to identify and clustering yeast cells of oral Candida species by CHROMagar test, SDS-PAGE and ELISA

2004 ◽  
Vol 64 (2) ◽  
pp. 317-326 ◽  
Author(s):  
J. A. de O. Rodrigues ◽  
J. F. Höfling ◽  
F. C. A. Tavares ◽  
K. M. R. Duarte ◽  
R. B. Gonçalves ◽  
...  
Keyword(s):  
Candida Species ◽  
Profile Analysis ◽  
Protein Profile ◽  
Negative Control ◽  
Yeast Cells ◽  
Sds Page ◽  
Differential Medium ◽  
Oral Yeasts ◽  
Species Specific ◽  
High Level

The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA), as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA), using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. Krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of cross-reaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S SM = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.

Sign in / Sign up

Export Citation Format

Share Document