scholarly journals The Potential of Candida Biofilm Protein as Bioreceptor for Candidiasis Immunoassay

2018 ◽  
Vol 14 (1) ◽  
pp. 47 ◽  
Author(s):  
Masfufatun Masfufatun ◽  
Loo Haryanto ◽  
Harsono Harsono

Abstract: Candidiasis or infection that is caused by Candida has become a new list of the therapeutical problems recently. The difficulties in diagnosing are the main cause of the unsatisfactory results from common therapies and diagnosis methods. This has urged researchers to find alternative ways in candidiasis diagnosis such as serology-based detection using antigen or antibody development. The aim of this study was to evaluate the potential of protein derived from Candida albicans biofilm as bioreceptor on candidiasis immunoassay through Dot Blot method. The research method used descriptive method with the following stages: (1) preparation of Candida albicans biofilm (2) extraction of Candida albicans protein through enzymatic and mechanical methods, (3) determination of protein molecular weight with SDS-PAGE (4) production of polyclonal anti- candida and (5) analysis of protein extract as bioreeceptor on dot blot. Profile of biofilm proteins on SDS-PAGE analysis were shown on molecular weight 27,42; 29,89; 38,10; 44,90; 48,75; 52,92; 55,14; 59,86; 70,56; 87,36; 102,54;115,05; 130,14;143,14;181,53 kD. There were differences in the intensity of dots in the control group (44070) and treatment groups (63170.5). It is noticeable that biofilm protein extract of C. albicans can be used for induction of anti-Candida polyclonal antibody production as the potential candidate of bioreceptor in candidiasis immunoassay. Keywords: SDS-PAGE, polyclonal antibody, immunoassay, dot blot, biofilm

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.


el–Hayah ◽  
2016 ◽  
Vol 5 (4) ◽  
pp. 169
Author(s):  
Sri Puji Astuti Wahyuningsih ◽  
Virid Gibson ◽  
Alfiah Hayati

<em>The purpose of this research was to determine the effect of polysaccharide krestin</em> (<em>PSK) </em><em>on the testicular protein profiles and testosterone levels of Mus musculus with variety of dosages. This research used a completely randomized design. It were devide into four treatment group i.e. control group, PSK treatment at a dose of  15, 30, 60 mg/kgBW. Each group had six replications. Testicular proteins were isolated by flushing technique and analized by SDS-PAGE. Testosterone levels were analized using ELISA technique at wavelength 450 nm. Protein bands analysis showed that there were no diversification between four treatments. Molecular weight of protein bands were 87, 63, 57, 35, and 30 kDa. The results of research showed that the testosterone levels at dosage 60 mg/kgBW had significanly different with control, PSK treatment of 15 and 30 mg/kgBW. PSK treatment of  60 mg/kgBW had lowest level at dosage, i.e. 25946.8 ρg/mL. It can be concluded that giving variety of dosages of polysaccharide krestin did not affect to testicular protein profiles but giving effect to testosterone levels of Mus musculus.</em>


2017 ◽  
Vol 18 (1) ◽  
pp. 25
Author(s):  
Mahful Septiawan ◽  
Budayatin Budayatin ◽  
Hidayat Teguh Wiyono ◽  
Kartika Senjarini

Although malaria had ever been virtually eradicated from Indonesia but currently malaria is recognized as a serious re-emerging threat to public health. This disease is caused by malaria parasite which is transmitted to human host by Anopheles mosquitoes as main vector. It has been widely observed that saliva of mosquito that transmits disease contains several factors that could enhance pathogen infection. Therefore, it should be possible to control pathogen transmission by vaccinating the host against the molecule(s) in saliva that potentiate the infection. However, immunogenic specific component in mosquitoes vectors of Malaria has not yet been identified so far. The objective of this study are to analyze protein profile of SDS-PAGE and to know the immunogity the protein extract of salivary gland from potential vector of Malaria i.e. An. aconitus We used immunogenic reaction between salivary gland extract of these vectors against pool of human sera which were collected from endemic area. The reaction conducted by the dot-blot analyze. SDS-PAGE studies showed 15 major polypeptide bands of 284, 100, 84, 75, 66, 57, 53, 48, 45, 38, 33, 29, 15, 14, and 11 kDa. The dot-blot studies showed that the protein extract of salivary gland from An. aconitus are immunogenic.


Author(s):  
Fahimeh ABDOLMALEKI ◽  
Zahra ZAMANI ◽  
Somayeh TALEBI

Background: Egg yolk is a rich and accessible source of yolk immunoglobulin (Y immunoglobulin). Presently, polyclonal antibodies from mammalian sources are used for diagnosis. Antibody production from egg yolk gives a higher yield and turnover than that from lab animals, and invasive methods such as phlebotomy and causing stress to the animals are not required. Due to the issues regarding mammalian antibodies, we aimed to evaluate the human anti-IgG polyclonal antibody production conjugated with peroxidase in egg yolk.Methods: Population of laying hens reared in Agriculture/Isfahan University of Technology were used in 2017. After immunizing hen against pure human IgG, specific IgY (yolk immunoglobulin) was purified from the yolk by sedimentation with polyethylene glycol (PEG6000). To assess the molecular weight and activity of the product, SDS-PAGE and ELISA-test were used, respectively.Results: The complete molecular weight of IgY was 180 kDa and the molecular weight of its light and heavy chains were 27 and 67 kDa, respectively.Conclusion: Antihuman IgG IgY had a purity above 90%. The product of this study can be used to measure IgG class antibodies in order to diagnose different diseases.


DENTA ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 48
Author(s):  
Anindita Apsari ◽  
Vivin Ariestania

<p><strong><em>Background: </em></strong><em>Heat cure acrylic and nylon thermoplastic valplast and lucitone-FRS brands are the three materials most often used as denture bases. Candida albicans is the dominant microorganism that can cause denture stomatitis. Cleaning denture can be done in two ways, namely the mechanical way with a toothbrush or an ultrasonic cleanser and a chemical method by immersing the denture into the cleaning solution. Chitosan solution is antibacterial and anti fungal which can inhibit the growth of Candida albicans colonies on heat cured acrylic plates, valplast and lucitone-FRS. <strong>Purpose:</strong> To determine the concentration of chitosan solution and the type of denture plate that most effectively inhibited the growth of Streptococcus mutans Candida albicans colonies on heat cured acrylic, valplast and lucitone-FRS plates. <strong>Materials and Methods: </strong>21 heat cured acrylic plate samples, 21 valplast plate samples, 21 lucitone-FRS plate samples measuring 10x10x2 mm divided by 9 groups. Heat cured, valplast and lucitone-FRS acrylic plate samples were contaminated with Candida albicans then immersed using the concentration of 0.25%, 0.5% chitosan solution and sterile aquades as the control group with 90 minutes immersion time. The entire study sample was calculated using Candida albicans colonies on Sabourroud's Dextrose Agar media. <strong>Results: </strong>The Kruskall Wallis test showed a significant difference (p &lt;0.05) in all treatment groups. Mann-Whitney showed a significant difference (p &lt;0.05) in all groups in the Candida albicans study. <strong>Conclusion: </strong>Soaking the lucitone-FRS plate in 0.5% chitosan solution for 90 minutes was the most effective in inhibiting the growth of Candida albicans colonies.</em></p><p><strong> </strong></p><p><strong><em>Keywords:</em></strong><em>Kitosan, Candida albicans, akrilik heat cured, valplast, lucitone-FRS</em></p><p><strong><em> </em></strong></p><p><strong><em>Correspondence:  </em></strong><em>Anindita Apsari, </em><em>Department of Prosthodontics, Faculty of Dentistry, Hang Tuah University,  Arif Rahman Hakim 150, Surabaya</em><em>, Email : </em><a href="mailto:[email protected]"><em>[email protected]</em></a><em>/</em><a href="mailto:[email protected]"><em>[email protected]</em></a><em></em></p>


2016 ◽  
Vol 49 (3) ◽  
pp. 148
Author(s):  
Muhammad Luthfi ◽  
Ira Arundina ◽  
Nizamiar Hamni

Background: Candida albicans (C. albicans) are dimorphic fungi in oral cavity, considered not only as normal flora, but also as pathogens. C. albicans have an ability to grow biofilm, which has a thick layer of outer skin structure, called as extracellular matrix. Jengkol leaves (Pithecellobium jiringa) contain alkaloids, flavonoids, terpenoids, and lectins, which have an ability as antifungal agent Purpose: This study aimed to analyze the optimum dose of jengkol leaf extract as antibiofilm against C. albicans biofilms. Method: C. albicans were cultured on yeast peptone dextrosa (YPD) media in 96-well microtiter plate flat bottom plates. There were one control group (without treatment) and three treatment groups. The first treatment group was given jengkol leaf extract at a dose of 100 mg/ ml. The second treatment group was given jengkol leaf extract at a dose of 200 mg/ ml. And, the third treatment group was given jengkol leaf extract at a dose of 400 mg/ ml. Semi quantitative method was applied to determine C. albicans biofilmsis using Crystal Violet staining technique. The absorbance of the cells then was calculated using a spectrophotometer with a wavelength of 570 nm. Result: The mean value of optical density in the control group was 1.23. The mean value of optical density in the treatment group with a dose of 100 mg/ ml was 0.2. Meanwhile, the mean value of optical density in the treatment group with a dose of 200 mg/ ml was 0.2, and 0.21 in the treatment group with a dose of 400 mg/ ml. The results also showed that there were significant differences between the control group and all of the treatment groups (p<0.05), but there was no significant difference between the treatment groups (p>0.05). Conclusion: The optimum dose of jengkol leaf extract used as antibiofilm against C. albicans biofilms is 100 mg /ml with an inhibitory percentage of 83.7%.


2019 ◽  
Vol 37 (1) ◽  
pp. 90
Author(s):  
Agung Janika Sitasiwi ◽  
Sri Isdadiyanto ◽  
Siti Muflichatun Mardiati

Abstract              Azadirachta indica (Neem) has been shown to affect the fertility of mice by interfering with the synthesis of testosterone in mice. The aim of this study was to detect the testes protein expression of mice after exposure to the ethanolic Neem leaf extract. The laboratory animals of this study were 20 male Swiss Webster mice with three months in age and body weight ranging from 27.5 grams. The mice were divided into two treatment groups, namely K (control group, exposed with distilled water) and P (treatment group, exposed to etahnolic Neem leaf  extract with 14 mg/animal /day). The treated were given for 21 days and the testicular protein was carried out on the 22nd day. The variables observed were testes weight, concentration and expression of proteins isolated from the testes. The protein concentration is determined by a spectrophotometer at a wavelength of 450nm. The protein expression was observed and determined based on the results of protein electrophoresis (SDS-PAGE). The results showed that protein expression in the treatment group has a lower concentration compared to the control group. Those results  was confirmed by thinner bands in SDS-PAGE result. Those proteins thought to be a fertility determinant in mammals. Keywords : anti-fertility; Neem; protein expression Abstrak  Azadirachta indica (Mimba) telah terbukti mempengaruhi fertilitas mencit dengan cara mengganggu sintesis hormon testoteron pada mencit. Penelitian ini bertujuan untuk melacak ekspresi protein pada testis mencit setelah paparan ekstrak etanol daun Mimba. Hewan uji penelitian ini adalah 20 ekor mencit Swiss Webster jantan dengan umur tiga bulan dan bobot badan berkisar 27.5 gram. Hewan uji dibagi menjadi 2 kelompok perlakuan, yaitu K (kelompok kontrol, dipapar akuades) dan  P (kelompok perlakuan, dipapar dengan ekstrak etanol daun Mimba dengan dosis 14 mg/ekor/hari). Pemberian bahan uji dilakukan  secara oral selama 21 hari. Variabel yang diamati adalah bobot testes, konsentrasi serta eskpresi protein yang diisolasi dari testis. Isolasi protein testis dilakukan pada hari ke-22. Konsentrasi protein ditentukan dengan spectrofotometer pada panjang gelombang 450nm. Ekspresi protein diamati dan ditentukan berdasar hasil elektroforesa protein. Hasil penelitian menunjukkan bahwa ekspresi protein pada kelompok perlakuan menunjukkan konsentrasi yang lebih rendah dengan pita yang lebih tipis jika dibandingkan dengan kelompok kontrol. Kesimpulan penelitian ini adalah paparan ekstrak etanol daun Nimba menyebabkan gangguan ekspresi protein yang diduga berperan dalam menentukan fertilitas mamalia. Kata kunci : Mimba; anti-fertilitas; eskpresi protein;   


2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Enny Suswati ◽  
Dian H. Purnamasari ◽  
Desie D. Wisudanti ◽  
Diana C ◽  
Mufida Mufida

Abstrak. Tujuan penelitian ini adalah membuktikan bahwa protein pili Shigella dysenteriae  dengan berat molekul 42 kDa merupakan protein adhesi dari S. dysenteriae pada enterosit mencit galur BALB/c. Penelitian ini merupakan penelitian eksperimental laboratoris yang dilaksanakan di Laboratorium Mikrobiologi Fakultas Kedokteran Universitas Jember, melalui beberapa tahap yaitu isolasi dan identifikasi S. dysenteriae, kultur S. dysenteriae, isolasi pili, SDS-PAGE, pemurnian protein pili, uji hemaglutinasi,isolasi enterosit mencit galur BALB/c, dan uji adhesi. Pada penelitian ini dibentuk 6 kelompok perlakuan dan 1 kontrol negatif. Keenam kelompok perlakuan tersebut meliputi konsentrasi protein pili 1, ½, ¼, 1/8, 1/16, dan 1/32. Data dianalisis menggunakan program statistik SPSS (Statistical Product and Service Solution) versi 16, jenis regresi linier sederhana dan one way Anov. Hasil penelitian menunjukkan bahwa protein pili S. dysenteriae 42 kDa mampu menghambat perlekatan S. dysenteriae terhadap enterosit mencit galur BALB/c. Pada uji regresi linier sederhana diperoleh nilai R square 0,897 yang berarti nilai indeks adhesi dipengaruhi oleh konsentrasi protein pili sebesar 89,7%, dan 10,3% dipengaruhi oleh faktor lain. Hasil uji one way Anova menunjukkan bahwa perbedaan konsentrasi protein pili berpengaruh terhadap indeks adhesi bakteri dengan nilai Sig. = 0,000 (p value 0,05). Kesimpulan yang diperoleh dari penelitian ini sesuai dengan hipotesis, yaitu protein pili dengan berat molekul 42 kDa merupakan protein adhesi dari Shigella dysenteriae pada enterosit mencit galur BALB/c. Kata kunci: Shigella dysenteriae,  protein pili  42 kDa, indeks adhesi Abstract. The aim of this study was to prove that the protein pili Shigella dysenteriae with a molecular weight of 42 kDa is an adhesion protein from S. dysenteriae in enterocytes of BALB / c mice. This research is a laboratory experimental research carried out at the Microbiology Laboratory of the Faculty of Medicine, University of Jember, through several stages, namely the isolation and identification of S. dysenteriae, culture of S. dysenteriae, isolation of pili, SDS-PAGE, purification of pili protein, hemagglutination test, isolation of enterocyte strain mice. BALB / c, and adhesion test. In this study, 6 treatment groups and 1 negative control were formed. The six treatment groups included protein concentrations of pili 1, ½, ¼, 1/8, 1/16, and 1/32. The data obtained were analyzed using the statistical program SPSS (Statistical Product and Service Solution) version 16, simple linear regression and one way Anov. The results showed that the 42 kDa S. dysenteriae pili protein was able to inhibit the attachment of S. dysenteriae to enterocytes of the BALB / c mice. In the simple linear regression test, it was obtained that the R square value was 0.897, which means that the adhesion index value was influenced by the pili protein concentration of 89.7%, and 10.3% was influenced by other factors. The one way Anova test results showed that the difference in pili protein concentration had an effect on the bacterial adhesion index with the Sig. = 0.000 (p value 0.05). The conclusion obtained from this study is in accordance with the hypothesis, namely pili protein with a molecular weight of 42 kDa is the adhesion protein of Shigella dysenteriae in enterocytes of BALB / c mice. Key words: Shigella dysenteriae, 42 kDa pili protein, adhesion index


2020 ◽  
Vol 72 (4) ◽  
pp. 1185-1196
Author(s):  
L.T. Patitucci ◽  
M.V. Azeredo ◽  
M.A. Verícimo ◽  
N.R.P. Almosny ◽  
M.C.N. Castro

ABSTRACT Glomerular proteinuria is characterized by the loss of high-molecular-weight proteins (HMWPs), while tubulointerstitial proteinuria is characterized by the loss of low-molecular-weight proteins (LMWPs). The objective was to assess the molecular weight of urinary proteins (MWUP) in dogs with naturally acquired CKD and determine the proportion of HMWPs and LMWPs according to CKD stage. Twenty-eight dogs with CKD were recruited and divided into 4 groups based on serum creatinine (Cr) levels (group1: Cr<1,4, n=8; group2: 1,4<Cr<2,0, n=6; group3: 2,1<Cr<5, n=9; group4: Cr>5,0, n=5). The control group consisted of 5 healthy dogs. The MWUP was determined by SDS-PAGE. The urinary protein-to-creatinine ratio (UP/C) was used to quantitatively assess proteinuria. The electrophoresis pattern revealed a proportionally greater loss of HMWPthan of LMWP in all groups with CKD and an increased loss of LMWP in group 4 (P<0.05). These results suggest a predominance of glomerular injuries throughout all stages of CKD in these dogs and an increase in tubulointerstitial injury towards the end-stage of the disease. The results of the present study support the recommendation of SDS-PAGE as an effective technique for the qualitative assessment of proteinuria, as well as a method for assessing the severity and location of renal injury.


e-GIGI ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 159
Author(s):  
Yuhi Syaula ◽  
Arlita L. Antari ◽  
Diah A. Purbaningrum

Abstract: Denture plate materials such as acrylic resin can induce adhesion of Candida albicans. Therefore, acrylic resin needs to be immersed in disinfectant. However, disinfectant can change its physical and mechanical properties, hence an alternative material is needed, such as hibiscus flower (Hibiscus rosa sinensis L.) due to its antifungal activity. This study was aimed to identify the effects of hibiscus flower extract and its concentrations towards the growth of C. albicans on acrylic resin plates. This was an experimental and laboratory study using the post-test only with control group design.  Acrylic resins were immersed in suspension of C. albicans, then were divided into four groups, as follows: 62.5% and 75% hibiscus flower extract (group I and II), positive control (sodium hypochlorite), and negative control (sterile aquadest). Acrylic resins were cultured and incubated on SDA media for 24 hours then the number of colonies were calculated. The results showed that C. albicans colonies in the treatment groups I, II, negative control, and positive control were 495 CFU/ml, 571.25 CFU/mL, 1175 CFU/mL, 23.125 CFU/mL respective-ly. The Kruskal-Wallis test showed significant differences in number of colonies of C. albicans (p<0.05) among all groups The post hoc Mann-Whitney test showed that all groups were significantly different, except for treatment groups I towards II. In conclusion, extract of hibiscus flower (H. rosa sinensis L.) affected the growth of C. albicans on acrylic resin plates.Keywords: hibiscus flower; Hibiscus rosa sinensis L.; Candida albicans; acrylic resin  Abstrak: Adanya bahan plat basis gigi tiruan seperti resin akrilik dapat memicu perlekatan C. albicans; oleh karena itu, resin akrilik perlu direndam dalam larutan desinfektan. Namun, larutan desinfektan dapat mengubah sifat fisik dan mekanik dari akrilik sehingga diperlukan adanya bahan alternatif, antara lain bunga sepatu (Hibiscus rosa sinensis L.) yang memiliki aktivitas antifungal. Penelitian ini bertujuan untuk mengetahui pengaruh perendaman ekstrak dan konsentrasi bunga sepatu terhadap pertumbuhan C. albicans pada plat resin akrilik. Jenis penelitian ialah eksperimental laboratorik dengan post-test only with control group design. Resin akrilik direndam dalam suspensi C. albicans, Terdapat empat kelompok perlakuan yaitu ekstrak bunga sepatu 62,5% dan 75%, kontrol positif (sodium hipoklorit), dan kontrol negatif (akuades steril). Resin akrilik dikultur dan diinkubasi pada media SDA selama 24 jam, kemudian jumlah koloni C. albicans dihitung. Hasil penelitian ini menunjukkan jumlah koloni C. albicans kelompok perlakuan I, II, kontrol negatif, dan positif sebanyak 495 CFU/ml, 571.25 CFU/mL, 1175 CFU/mL, 23.125 CFU/mL secara berurut. Uji Kruskal-Wallis menunjukkan perbedaan jumlah koloni C. albicans yang bermakna (p<0.05) antar semua kelompok. Uji post hoc Mann-Whitney menunjukkan semua kelompok berbeda bermakna, kecuali kelompok perlakuan I dengan II. Simpulan penelitian ini ialah ekstrak bunga sepatu (H. rosa sinensis L.) berpengaruh terhadap pertumbuhan C. albicans pada plat resin akrilik.Kata kunci: bunga sepatu; Hibiscus rosa sinensis L.; Candida albicans; resin akrilik


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