Immunogenity of Protein Extract from Salivary Gland of Anopheles aconitus in Malaria Endemic Area

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.

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INDIVIDUAL HUMAN SERA RESPONSE AGAINST PROTEIN EXTRACTS FROM SALIVARY GLAND OF Aedes aegypti

KnE Life Sciences ◽

10.18502/kls.v2i1.121 ◽

2015 ◽

Vol 2 (1) ◽

pp. 86

Author(s):

Rike Oktarianti ◽

Kartika Senjarini ◽

Fatchiyah . ◽

Aulani’am .

Keyword(s):

Salivary Gland ◽

Aedes Aegypti ◽

Complex Mixture ◽

Biologically Active ◽

Dengue Viruses ◽

Sds Page ◽

Human Sera ◽

Immunogenic Proteins ◽

Hematophagous Arthropods ◽

Individual Human

The saliva of hematophagous arthropods contains a complex mixture of biologically active proteins. These proteins may modify hemostatic responses and induce both cellular immunity and the production of specific antibodies, and thus influence the transmission of its pathogens from arthropods vector to human host. Aedes aegypti is the main vector for transmission of dengue viruses into human. The objective of this study was to examine individual human sera response against protein extracts from salivary gland of Ae aegypti that mediate the infection of dengue viruses. We did a cross reaction test of human sera from healthy people in endemic and non-endemic area, and dengue patients againts SGE of Ae. aegypti to distinguish and to identify the immunogenic proteins using Western Blot Analysis. About 15 protein bands of SGE from Ae. aegypti ranging from 15 kDa up to 255 kDa were identified on 12% SDS-PAGE. Seven dominant bands were detected, i.e ~255, 56, 42, 31, 27, 26 and 15 kDa. Two immunogenic proteins, as represented by two bands, i.e. ~31 and 56 kDa were found only in samples from people who were previously exposed to mosquitoes bites, and not in people who had not been exposed. Therefore, these immunogenic salivary proteins may serve as indicators for the immune response in human against protein from salivary gland of Ae. aegypti.Keywords: immunogenic proteins, salivary gland, Aedes aegypti

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The Potential of Candida Biofilm Protein as Bioreceptor for Candidiasis Immunoassay

Berkala Kedokteran ◽

10.20527/jbk.v14i1.4539 ◽

2018 ◽

Vol 14 (1) ◽

pp. 47 ◽

Cited By ~ 1

Author(s):

Masfufatun Masfufatun ◽

Loo Haryanto ◽

Harsono Harsono

Keyword(s):

Molecular Weight ◽

Candida Albicans ◽

Polyclonal Antibody ◽

Control Group ◽

Potential Candidate ◽

Dot Blot ◽

Protein Extract ◽

Treatment Groups ◽

Sds Page ◽

Mechanical Methods

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

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PURIFIKASI PROTEIN IMUNOGENIK 31 DAN 56 kDa DARI KELENJAR SALIVA Aedes aegypti

Jurnal Bioteknologi & Biosains Indonesia (JBBI) ◽

10.29122/jbbi.v7i1.3931 ◽

2020 ◽

Vol 7 (1) ◽

Author(s):

Syubbanul Wathon ◽

Fitria Mutiah ◽

Rike Oktarianti ◽

Kartika Senjarini

Keyword(s):

Salivary Gland ◽

Aedes Aegypti ◽

Positive Control ◽

Protein Fractions ◽

Immunogenic Protein ◽

Dot Blot ◽

Molecular Weights ◽

Sds Page ◽

Salivary Gland Protein ◽

Immunogenic Proteins

Purification of 31 and 56 kDa Immunogenic ProteinsÂ from the Salivary Glands of Aedes aegyptiThe salivary gland of arthropod vector contains various bioactive compounds and plays a role in the transmission of pathogens to the host. The host develops anti-salivary antibodies against vector saliva exposure. Our previous research has identified two immunogenic proteins with molecular weights of 31 and 56 kDa from the Aedes aegypti salivary gland protein extract. However, the role of the 31 and 56 kDa immunogenic proteins from saliva Ae. aegypti is not fully known, so it is necessary to purify two immunogenic protein fractions to better specify the target of developing a dengue vaccine. This study aimed to purify the 31 and 56 kDa immunogenic protein fractions by electroelution and dialysis methods. The purification of the two protein fractions has been successful which were confirmed by the SDS-PAGE by the existence of single-band parallel to the positive control. These results were further supported by the dot blot analysis which showed a positive reaction in the form of dark spots in the two protein fractions which were reacted with dengue patients' serum, endemic healthy people, and neonates. These results indicated that the purified 31 and 56 kDa immunogenic protein fraction can be identified by specific antibodies.Keywords: dialysis, electroelution, immunogenic, purification, saliva Â ABSTRAKKelenjar saliva vektor arthropoda mengandung berbagai senyawa bioaktif dan berperan dalam transmisi patogen ke tubuh inang. Tubuh inang mengembangkan antibodi anti-saliva terhadap paparan saliva vektor. Penelitian kami sebelumnya telah mengidentifikasi dua protein imunogenik dengan berat molekul 31 dan 56 kDa dari ekstrak protein kelenjar saliva Aedes aegypti. Namun demikian, peranan protein imunogenik 31 dan 56 kDa dari saliva Ae. aegypti belum diketahui sepenuhnya sehingga perlu dilakukan purifikasi dua fraksi protein imunogenik untuk lebih menspesifikkan target pengembangan vaksin dengue. Tujuan penelitian ini untuk melakukan purifikasi fraksi protein imunogenik 31 dan 56 kDa melalui metode elektroelusi dan dialisis. Keberhasilan purifikasi dua fraksi protein 31 dan 56 kDa terbukti dari hasil konfirmasi SDS-PAGE dengan terbentuknya pita tunggal sejajar dengan kontrol positif. Hasil tersebut diperkuat dengan analisis dot blot yang menunjukkan reaksi positif dengan munculnya noktah gelap pada dua fraksi protein tersebut ketika direaksikan dengan serum pasien DBD, penduduk sehat endemik dan neonatus. Hasil ini mengindikasikan bahwa fraksi protein imunogenik 31 dan 56 kDa hasil purifikasi dapat dikenali oleh antibodi spesifik.

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In vitro analysis of human immune response (IgG) against salivary gland extract of dengue vector from dengue hemorrhagic fever (DHF) endemic area in Jember, Indonesia

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/913/1/012090 ◽

2021 ◽

Vol 913 (1) ◽

pp. 012090

Author(s):

R Oktarianti ◽

D R Damara ◽

S U R Qudsiyah ◽

S Wathon ◽

K Senjarini

Keyword(s):

Immune Response ◽

Salivary Gland ◽

Endemic Area ◽

Mosquito Species ◽

Host Immune System ◽

Salivary Gland Extract ◽

Gland Extract ◽

Human Immune Response ◽

Human Immune

Abstract The mosquito species Ae. aegyptiand Ae. albopictusare two potential vectors of dengue fever. The salivary glands of these species contain substances that play a role in the transmission of pathogens. These include vasodilators and immunomodulatory compounds. Immunomodulatory components can modulate the host immune system by producing specific antibodies (IgG). This study aims to investigate the human immune response (IgG) against the salivary gland extract of Ae. aegyptiand Ae. albopictus. Samples were collected from individuals who were Dengue patients, as well as healthy individuals and neonates from the Jember endemic area. Results show that the levels of IgG response vary across the individual. Generally, Dengue patients and healthy people in the DHF-endemic area had higher levels of IgG. The highest immune response was found in DHF patients, followed by healthy persons, and finally the neonate samples, respectively.

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Revealing Antioxidant and Antidiabetic Potency of Melinjo (Gnetum Gnemon) Seed Protein Hydrolysate at Different Stages of Seed Maturation

Current Research in Nutrition and Food Science Journal ◽

10.12944/crnfsj.7.2.17 ◽

2019 ◽

Vol 7 (2) ◽

pp. 479-487 ◽

Cited By ~ 2

Author(s):

Anang Supriyadi ◽

Laras Sekar Arum ◽

Ari Satia Nugraha ◽

Anak Agung Istri Ratnadewi ◽

Tri Agus Siswoyo

Keyword(s):

Seed Protein ◽

Protein Hydrolysate ◽

Protein Profile ◽

Radical Scavenging ◽

Free Radical Scavenging ◽

Antidiabetic Activity ◽

Degree Of Hydrolysis ◽

Protein Extract ◽

Sds Page ◽

Gnetum Gnemon

roteins hydrolyzed from melinjo seeds (Gnetum gnemon) at green (GM), yellow (YM) and red (RM) stages of maturity were studied for their effectiveness in antioxidant and antidiabetic activities. The seed protein extract was hydrolyzed using alcalase 2.4L, and the resulting hydrolysates with the highest degree of hydrolysis, protein profile, and the most potent contributors to antioxidant and invitro antidiabetic activities were identified. The degree of hydrolysis value of hydrolysates ranged from 52-84%, and the SDS-PAGE protein profile showed two distinct bands in which the band with molecular weight of 30 kDa degraded more intensively. Antioxidant capacity was measured using different standard methods, including radical cation 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS•+) assay, hydroxyl radical (OH•), and superoxide anion (O2•-) scavenging. The green hydrolysate (GMH) had significantly higher (p<0.05) free radical scavenging (ABTS•+, OH•, and O2•-) activities than that of the yellow hydrolysate (YMH) and red hydrolysate (RMH). However, invitro antidiabetic testing was performed based on the inhibitory activity of α-amylase and α-glucosidase. GMH was found to be more effective than YMH and RMH. These results showed that the antioxidant and antidiabetic activity in hydrolyzed GM protein has high potential to be utilized as natural nutraceuticals.

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Successful interrupted feeding of adult Rhipicephalus appendiculatus (Ixodidae) is accompanied by re-programming of salivary gland protein expression

Parasitology ◽

10.1017/s0031182099004540 ◽

1999 ◽

Vol 119 (2) ◽

pp. 143-149 ◽

Cited By ~ 22

Author(s):

H. WANG ◽

P. J. HENBEST ◽

P. A. NUTTALL

Keyword(s):

Salivary Gland ◽

Protein Expression ◽

Protein Profile ◽

Rhipicephalus Appendiculatus ◽

Pathogen Transmission ◽

Ixodid Ticks ◽

Egg Mass ◽

New Host ◽

Body Weights ◽

Salivary Gland Protein

Ixodid female ticks take one comparatively large bloodmeal which they convert to a single large egg mass and then they die. To examine the outcome of interrupted feeding, equal numbers of male and female Rhipicephalus appendiculatus adult ticks were fed on guinea pigs (host 1) for either 2, 4, or 6 days, or to engorgement (8 days). All of the fully engorged (D8) females laid a single large egg mass (80–160 mg/tick), while 85% of the day 6-fed (D6) female ticks (n=20) each laid a small egg mass (6·1 mg/tick). None of the females that had fed for 2 or 4 days oviposited. Ninety percent (n=20) of the day 2-fed (D2) females survived for 4 weeks after their feeding was interrupted, whereas 65% (n=20) of the day 4-fed (D4) females survived. All of the surviving partially fed female ticks (D2 and D4) attached to a second guinea pig (host 2) and attained engorged body weights that were not significantly different from those of the control females (P<0·05). Female ticks that engorged following interrupted feeding layed egg masses comparable to the controls, indicating that engorgement on host 2 was successful. The salivary gland protein profile of female ticks changed constantly during feeding. However, when feeding was interrupted, the protein expression pattern switched back to that of the non-parasitic state, presumably to enable the partially fed ticks to survive and reattach on the new host. This observation indicates that female ixodid ticks have a natural ability to survive and re-establish successful feeding on a new host if the first attempt at feeding is unsuccessful. Such an interrupted feeding mechanism supports the hypothesis that partially engorged ticks may play a role in tick-borne pathogen transmission.

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Isolation and Structural Charaderization of a Potent Inhibitor of Coagulation Factor Xa from the Leech Haementeria ghilianii

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646610 ◽

1989 ◽

Vol 61 (03) ◽

pp. 437-441 ◽

Cited By ~ 28

Author(s):

Cindra Condra ◽

Elka Nutt ◽

Christopher J Petroski ◽

Ellen Simpson ◽

P A Friedman ◽

...

Keyword(s):

Molecular Weight ◽

Salivary Gland ◽

Amino Acid ◽

Amino Acid Sequence ◽

Western Blot ◽

Potent Inhibitor ◽

Factor Xa ◽

Coagulation Factor ◽

Sds Page ◽

Bovine Factor

SummaryThe present work reports the discovery and charactenzation of an anticoagulant protein in the salivary gland of the giant bloodsucking leech, H. ghilianii, which is a specific and potent inhibitor of coagulation factor Xa. The inhibitor, purified to homogeneity, displayed subnanomolar inhibition of bovine factor Xa and had a molecular weight of approximately 15,000 as deduced by denaturing SDS-PAGE. The amino acid sequence of the first 43 residues of the H. ghilianii derived inhibitor displayed a striking homology to antistasin, the recently described subnanomolar inhibitor of factor Xa isolated from the Mexican leech, H. officinalis. Antisera prepared to antistasin cross-reacted with the H. ghilianii protein in Western Blot analysis. These data indicate that the giant Amazonian leech, H. ghilianii, and the smaller Mexican leech, H. officinalrs, have similar proteins which disrupt the normal hemostatic clotting mechanisms in their mammalian host’s blood.

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Rhipicephalus sanguineus salivary gland extract as a source of immunomodulatory molecules

Experimental and Applied Acarology ◽

10.1007/s10493-021-00591-w ◽

2021 ◽

Author(s):

Melissa Carolina Pereira ◽

Elen Fernanda Nodari ◽

Marina Rodrigues de Abreu ◽

Lisiery Negrini Paiatto ◽

Patrícia Ucelli Simioni ◽

...

Keyword(s):

Salivary Gland ◽

Rhipicephalus Sanguineus ◽

Salivary Gland Extract ◽

Gland Extract

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Comparative Analysis of Electrophoretic Profile of Major Proteins of Milk from Alpine and Carpathian Goats

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Veterinary Medicine ◽

10.15835/buasvmcn-vm:12447 ◽

2017 ◽

Vol 74 (1) ◽

pp. 20 ◽

Cited By ~ 1

Author(s):

Alina NASALEAN ◽

Laurentiu OGNEAN ◽

Sergiu MUNTEAN ◽

Stefana BALICI ◽

Horea MATEI

Keyword(s):

Protein Profile ◽

Milk Proteins ◽

Goat Milk ◽

Protein Composition ◽

Raw Milk ◽

Biologically Active ◽

Protein Fractions ◽

Animal Nutrition ◽

Sds Page ◽

Percentage Data

The milkâ€™s proteins provide nutritional and biologically active values, essential in human and animal nutrition. In the case of goat milk, the proteinsâ€™ concentration and quality represent basic indices for the evaluation of the nutritional and biologically active values. The proposal is to comparatively analyse the protein profile of milk. The milk was collected from two different breeds: French Alpine and Romanian Carpathian. During March and April 2016 there were collected samples of raw milk in hygienic and sanitation conditions. There were two lots: first lot has 10 Carpathian goats and the second lot has 10 Alpine goats. The protein composition of goat milk was established with SDS-PAGE, after the evaluation of the total proteinsâ€™ concentration with the Bradford method. The quantitative and percentage data obtained with electrophoresis revealed few differences between those 8 identified protein fractions. Between those two lots, regarding the levels of Î²-CN, k-CN and Î²-lactoglobulines there were significant differences. The other protein fractions have values almost identical. Statistical analysis of obtained data shaped the differences in the protein profile at those two breeds. Based on those differences it is to note the superior potential of the Alpine breed regarding the content in biologically active milk proteins. Regarding the obtained data, this study brings new contributions for the evaluation and analysis of protein profile as a nutritive and biologically active component of goat milk, confirming its character as a functional aliment.

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