Characterisation of oyster allergens for improved diagnosis of mollusc allergy

Discovery and characterisation of allergens in various food and inhalant sources is central to our understanding of the molecular mechanisms of allergic reactions. Allergen characterization is the most important underlying factor for better patient management with improved diagnostics, and the design and development of novel immunotherapeutics. Of the 'Big eight' allergen food groups, shellfish presents a unique challenge in terms of allergen discovery due to the large number and diversity of consumed species, leading to heterogeneity of allergen structure and cross-reactivity among various sources. The group of 'shellfish' comprises of two invertebrate phyla; arthropods and molluscs. Although all shellfish are invertebrate animals, these two groups are very distinct in evolutionary terms and subsequently contain different molecular repertoires of allergenic proteins. Co-sensitisation of patients with crustacean and mollusc allergy is often described, however, the current diagnostic approaches to manage patients is not based on sufficient molecular knowledge of these shellfish allergens. Consequently, mollusc allergy is clinically underreported and allergens are ill-defined. To date, only five mollusc allergens are listed in the WHO/International Union of Immunological Societies (IUIS) Allergen Nomenclature SubCommittee database, all of which are different tropomyosin's (http://www.allergen.org/index.php). Additional mollusc allergens have been reported, but not yet fully characterised. A detailed review of the current status and the diagnosis of mollusc allergy is provided in Chapter 1. Current strategies for allergen identification are time- and resource-consuming, which are highly prone to missing hidden allergens present in low concentrations. Allergenic proteins are traditionally identified based on their serum-specific IgE antibody recognition. Soluble proteins derived from whole protein preparations of a suspected allergen source are screened for IgE antibody binding proteins using sera from individuals with clinically confirmed allergy. Although this approach is the current standard for allergen identification, there are three major drawbacks. Firstly, this approach often does not detect allergenic proteins present in low abundance. Secondly, cross-reactive allergens are not easily identified due to their possible presence in unrelated allergen sources. Thirdly, the IgE recognition patterns are highly dependent on the demographics of the particular allergic patient cohort under investigation. This PhD thesis presents a comprehensive study on the improvement of allergen discovery from Pacific oyster, the most widely consumed mollusc species and immunological characterisation of the major allergen, tropomyosin, using a cohort of mollusc-sensitised patients in Australia. Furthermore, novel ways to diagnose cross-reactivity between crustacean and mollusc species were developed. Chapter 2 describes a comprehensive discovery pipeline, for allergenic proteins, that accounts for biological and molecular variability using allergenomics, high-throughput screening of genomic databases and high-resolution mass spectrometry. This methodological approach was successful in identifying 24 previously unreported allergens from over 25,000 proteins from the Pacific oyster. This is the first study to demonstrate the presence of 24 hidden allergens, also found in very different allergen sources from animals, including fish and mites, as well as plant allergens from pollen, latex and fungi. Importantly all of these allergenic proteins identified are reactive with shellfish allergic patients' IgE antibodies. However, it was demonstrated in chapter 2 that not all allergens present in the genome and transcriptome of oyster are also detected in the extracted proteome. Allergens are often overlooked during the extraction process due to the use of inappropriate buffers which might affect the in vitro diagnostic methods that use whole proteins extracts. In Chapter 3 various buffer compositions covering a wide range of pH were evaluated to improve the detection of the unreported allergens described in chapter 2. The IgE-reactivity of protein extracts from each buffer was determined against a pool of serum from five shellfish allergic patients. In addition, the protein composition of the Pacific oyster was analysed using high-resolution mass spectrometry. High concentrations of protein were recovered after extraction using high salt content or high pH buffers, subsequently revealing more IgE-reactive bands on the immunoblotting. Low pH buffers, however, resulted in poor protein recovery and affected negatively patient IgE-reactivity. Mass spectrometry analysis discovered that the novel IgE-reactive proteins, particularly of high molecular weight, emerged due to an increased abundance in the allergen extract. Overall, increasing the ionic strength and pH of the extraction buffers improves the solubility of allergenic proteins. In Chapter 4, a detailed analysis of immunological characteristics of the Pacific oyster extracts and the major allergen, tropomyosin were conducted. Twentyone oyster-sensitised patients were analysed to determine the prevalence of each allergen in the patient cohort. Eighteen out of 21 patients showed reactivity to tropomyosin although the binding intensity varied between patients. Patients who lacked IgE-binding to tropomyosin were shown to be sensitised to other oyster allergens. These allergenic proteins were preliminary abundant in the raw extract of the Pacific oyster. Further investigation was carried out with the Pacific oyster tropomyosin. The natural wild type and recombinant tropomyosin were successfully purified, and their structural properties observed. Both purified natural and recombinant tropomyosin had very similar structural and immunological properties. Cross-reactivity analysis using ELISA demonstrated patients who demonstrated IgE reactivity to the Pacific oyster tropomyosin were also reactive to other tropomyosin's from Black tiger prawn as well as the House dust mite. The degree of crossreactivity correlated well with the tropomyosin amino acid sequence identity. Sequence alignment of tropomyosin from those three species revealed five protein regions containing predicted IgE-binding epitopes responsible for the strong cross-reactivity observed. Finally, in Chapter 5 to improve the prediction of clinical cross-reactivity between crustaceans and molluscs as well as other invertebrate species, conservation analysis of IgE-binding epitopes of four shrimp allergens were carried out. The results demonstrated that within a large directory of shrimp IgE-binding epitopes there are a substantial fraction of epitopes that are highly conserved across various invertebrate species. Shrimp TM and AK shared a higher number of conserved epitopes compared to shrimp SCP and MLC; in fact, no conserved epitope could be found for SCP, while MLC only shared two epitopes in one region with cockroach MLC. These results suggest that TM and AK are the major contributing proteins in immunological and clinical cross-reactivity between crustacean and other invertebrate groups. Furthermore, comparative evaluation of the number of conserved epitopes in TM and AK revealed a clear cross-reactive hierarchy where cockroach has the highest number followed by mite, and molluscs are on the bottom of this hierarchy. In conclusion, the outcomes of this thesis have demonstrated that many unreported allergens have been overlooked due to the limitations of the current allergen discovery methodology. The utilisation of transcriptome data and proteomic techniques in addition to the well-established allergenomic approach improves discovery of unreported allergens. While tropomyosin seems to be a clinically relevant cross-reactive major allergen, the presence of additional allergens that has never been reported in mollusc species, suggests mollusc species contain different molecular repertoires of allergens. Supported by the findings of the in-depth bioinformatics analysis of IgE-binding epitopes, component-resolved diagnostics for mollusc allergy could be developed, enabling precise identification of patients sensitised to a specific mollusc group and distinguish from patients with extensive cross-reactivity to ingested and inhaled allergens from other invertebrate sources

MEANING FIELD THEORY IN CELEBGRAM ENDORSEMENT PRODUCT CAPTIONS OF IKA PURPITASARI

Manufacturers of a product definitely want their products to be bought and used by the public. Everything is done by the marketing department of the product, including online marketing through social media. It is considered more effective and efficient. In Indonesia, the development of the online world continues to grow rapidly until now. Many online social media are used by Indonesian people, one of which is Instagram. Instagram is one of the favorite social media, sometimes termed celebgram or celebrities instagram.  Celebrities are chosen by product manufacturers to boost sales of their products in society. Celebgram with a fairly large number of followers are considered capable of becoming an online marketing agent. In this study, the researcher analyzes the caption of the endorsement of photo product from a celebrity named Ika Purpitasari. When analyzing the captions, the researcher uses the field of meaning theory derives from the semantic discipline. The photo captions analyzed by the researcher are those that contain endorsement products in the field of womanhood, such as women's fashion, women's beauty, and women's accessories. The  research method is descriptive analysis using the data collection techniques of documentation. The results of this study show that the field of meaning theory can be used outside of spoken language, namely when analyzing the captions in the photos of the celebgram.

THE USE OF BLENDED LEARNING THEORY IN NEGOTIATION TEXT MATERIAL IN HIGH SCHOOL

The background of this research is because I was inspired by the pandemic conditions that hit the country. Students often confide in the researcher. They want material that is contemporary,contextual, not merely textual. Therefore, the researcher took one of the materials about the negotiation text combined with the blended learning theory. The negotiation text that the researcher used was a text about conversation in one hospital, which was sourced from an online news site.The method that researcher use in this research is descriptive analysis. The researcher took the blended learning literature by Wasis D Dwiyogo. The results of this study indicate that blended learning is beneficial when it is translated into negotiation text material. Last but not least, the conclusion of this research shows that blended learning is beneficial when combined with negotiation text material

Reformist Muslim Discourse in the Sundanese Commentary of the Qur’ān: E. Abdurrahman’s Commentary on QS. Al-Hujurāt

This article focuses on reformist Muslim discourse in the Sundanese commentary of the Qur’ān or tafsīr, entitle Sababaraha Naséhat Tina Qur’an Surat al-Hudjurot, some advice of sūrah Al-Hujurāt (1971). The book was written by E. Abdurrahman (1912-1983), an activist of a reformist Islamic organization, Islamic Union (Persatuan Islam or Persis). He has been a chairman of Persis from 1962-1983. They have a slogan “back to the Qur’ān and Hadīth” as well as to oppose the taqlīd and heresy. E. Abdurrahman explained Q. 49 or sūrah Al-Hujurāt in the frame of his ideology as an activist of Persis. Following his commentary of Q. 49:1, the prohibition against practicing Islam before find dalīl or textual instructions of the Qur’ān and Hadīth. He also interpreted Q. 49:6 on recheck the news as a form of prohibition taqlīd. Meanwhile, Q. 49:9-10 was interpreted by him to emphasizes the strengthening of reformist followers. Using Fairclough’s critical discourse analysis, this study confirms that E. Abdurrahman’s commentary cannot be separated from his reformist ideology. He used Sundanese Quranic commentary to convey the reformist ideas against traditionalist Muslims in West Java

Implementasi Sistem Check and Balances dalam Keadaan Bahaya Menurut Sistem Ketatanegaraan Republik Indonesia

Keadaan bahaya di beberapa wilayah Indonesia sering terjadi, seperti pemberontakan, perang sipil, bencana alam, konflik sosial, ekonomi, pertahanan, dan keamanan. Masing-masing keadaan bahaya terjadi dengan jenis keadaan yang berbeda serta skala yang besar, sehingga menimbulkan banyak korban jiwa. Dalam penanganan suatu keadaan bahaya negara Indonesia harus dilaksanakan dengan sistem check and balances. Tujuan dari penelitian ini adalah mengetahui dan menganalisis pengaturan keadaan bahaya dalam sistem ketatanegaraan Republik Indonesia dan implementasi sistem check and balances dalam keadaan bahaya menurut sistem ketatanegraan Republik Indonesia. Penelitian ini menggunakan metode pendekatan yuridis normatif dengan spesifikasi penelitian deskriptif. Sumber data dari data sekunder berupa studi pustaka. Penelitian ini diuraikan dengan teks naratif yang dianalisis dengan metode analisis kualitatif. Berdasarkan hasil penelitian dan pembahasan maka dapat disimpulkan bahwa bahwa dalam pengaturan mengenai keadaan bahaya bentuk pembelaan diri negara lebih menonjol segi kemiliteran. Pihak militer sebagai stabilisator maju ke depan untuk menghadapi keadaan bahaya dan ancaman-ancaman untuk memulihkan keadaan bahaya menjadi normal kembali. Sistem check and balances dalam keadaan bahaya tidak diterapkan dengan optimal, kewenangan presiden yang sangat luar biasa tanpa ada pembatasan tanpa ada pengawasan langsung dari satu cabang pemerintahan terhadap cabang pemerintahan lainnya

Analisis Proses Morfologis Afiksasi Pada Berita Media Online Tribunnews

Penelitian ini dilaksanakan karena ditemukan artikel berita menarik di Tribunnews. Penelitian ini adalah penelitian dengan menggunakan pendekatan kualitatif untuk mengolah data yang akan dianalisis berdasarkan teori yang menjadi rujukan. Permasalahan penelitian berada pada media Tribunnews dengan berfokus pada morfologi dalam proses afiksasi. Tujuan penelitian ini adalah untuk mengetahui deskripsi proses morfologis afiksasi dalam berita media online Tribunnews. Metode yang digunakan dalam penelitian ini menggunakan metode analisis isi. Teknik analisis data pada penelitian ini, yaitu: (1) Identifikasi data, ini dilakukan untuk mengetahui pembentukan kata yang ada dalam berita media online Tribunnews. (2) Setelah diidentifikasi data selanjutnya mengklasifikasi setiap bentuk penggunaan kata berdasarkan jenisnya. (3) Kemudian data yang sudah diklasifikasi selanjutnya dideskripsikan sesuai dengan fungsi dan makna proses morfologis afiksasi. Adapun judul berita, yaitu: “Fenomena Parade di Langit Bakal Terjadi Besok, Bisa Disaksikan dari Wilayah Indonesia”, “Sejarah Kapitan Pattimura alias Thomas Matulessy, Pahlawan Maluku di Era Penjajahan Belanda”, “Perubahan Angin Taifun Menjadi Siklon Ekstratropis, Tokyo Jepang Diprediksikan Hujan Deras Malam Ini”, “Terungkap! Sampah Sesat paling Banyak Cemari Perairan Bali, Produsen Diminta Lakukan Ini”, dan “Gempa Magnitundo 5,3 Guncang Bengkulu Siang Ini”

SISTEM INFORMASI UJIAN SEKOLAH ONLINE BERBASIS DATA WAREHOUSE (STUDY KASUS : SMP NEGERI 2 LURAGUNG)

Web based information system is in great demand in various circles, including in educational institutions, especially in institutions that have not been computerized, written test system that is often used in academic activities often have many obstacles. The cheating factor and the habit of cheating is the biggest obstacle, besides the factor of the slow assessment process in written test due to the many participants that can hamper the teacher in putting the value into the report card, waste paper and ink cost in making problem and doubling the problem including into negative factor in written exam, this study aims to create an online Data Warehouse based test system that can be utilized in educational institutions to change the current written exam into an online exam system. Therefore, in line with the rapid development of information technology, it is necessary to design and build an Online Data Warehouse based test system to replace the manual written test system. Where this system is expected to reduce the negative factors that often occur, as well as facilitate the teachers in SMP Negeri 2 Luragung in input data into the system, and can minimize operational costs in conducting exams in schools, as well as systems that will be made capable of storing data in large amounts and more secure in data storage, such as teacher data, principal data, student data, subject data, value data and data question.The method used is Data Warehouse, software development procedure using Waterfall method. The research respondents are the students of class IX in SMP Negeri 2 Luragung. Online test system is created by using PHP programming language, MySQL as database and XAMPP web serve

Analisis Tindak Tutur Perlokusi di Toko Alat Rumah Tangga Pasar Cibarusah Kabupaten Bekas

Penelitian ini berkenaan dengan tindak tutur perlokusi pedagang di toko Aneka Ilham Cibarusah dalam tindak tutur perlokusi yang dibagi menjadi dua jenis, diantaranya tindak tutur perlokusi verbal dan tindak tutur perlokusi verbal nonverbal. Tujuan penelitian untuk mendeskripsikan wujud tuturan, efek, dan fungsi tindak tutur perlokusi. Metode penelitian ini berpijak pada metode penelitian deskriptif, data yang diperoleh bukanlah berupa angka melainkan kata-kata dan kalimat.  Hasil penelitian bentuk tindak tutur perlokusi verbal  yang terdapat pada tuturan transaksi jual beli di Aneka Ilham. Informan Ida Deriyanti menuturkan lima tindak tutur perlokusi verbal. Informan Ujang Juhaedi menuturkan enam tindak tutur perlokusi verbal. Bentuk tindak tutur perlokusi verbal nonverbal yang terdapat pada tuturan transaksi jual beli di toko Aneka Ilham. Informan Ida Deriyanti menuturkan sepuluh tindak tutur perlokusi verbal nonverbal. Informan Ujang Juhaedi menuturkan enam tindak tutur perlokusi verbal nonverbal. Hasil penelitian ini dapat digunakan untuk menambah wawasan pengetahuan atau referensi untuk orang lain

THE EXPRESSION OF PIETY THROUGH THE RATIK TOGAK IN THE TAREKAT SURAU SULUK IN RIAU

Tarekat is piety expression based on spirituality encouragement as Islamic identity in Indonesia. Studies about piety expression is often connected to redicalism or violent movement. In fact, piety expression of Muslims in Indonesia as described in this article can accomodate with local culture. This study examines the existence of Surau Suluk and the tradition of Ratik Togak, which is one of the practices of the Naqsabandiyah tarekat in Rokan Hulu. It is known ‘Negeri Seribu Suluk’, having approximately 122 Surau Suluk. Surau Suluk is a place to undergo religious activities to get closer to God. This study uses qualitative descriptive method employing observation and interview. A phenomenological concept is used to see the religious traditions of followers of the Surau Suluk congregation. The results showed that Surau Suluk in Rokan Hulu is developed by Sheikh Abdul Wahab Rokan. A practice that has become a tradition in the Naqsabandiyah tariqah is Ratik Togak. Ratik means dhikr, a practice in the form of dhikr and forgiveness which is done to find the peak of enjoyment of dhikr. The Togak Ratik is performed at certain religious events in public spaces as an important medium for them to get closer to God. Ratik Togak is a form of social Islamic religious worship. This tradition is special because it has become an icon for Surau Suluk which involves ordinary people. It also gets support from local government. This tradition is a practice for followers of the Naqsabandiyah as a form of their piety with God

PENGARUH PENGGUNAAN INTERNET TERHADAP KESADARAN POLITIK (KASUS PILKADES 2019 DI DESA CICADAS, KACAMATAN GUNUNGPUTRI, KABUPATEN BOGOR)

This research was conducted to analyze about the effect of internet use on political awareness. This study uses one independent variable, namely internet use and one dependent variable, political awareness. The sampling technique used was Non-probability sampling with accidental sampling technique. The data analysis method used is a quantitative analysis of the validity and reliability test, the classic assumption test, simple linear regression analysis, T test and F test and the coefficient of determination using SPPS software version 22 for windows. The results showed the points of the indicator variables X are intensity, usefulness, effectiveness and variable Y which consists of indicators of knowledge, understanding, attitudes, and patterns of behavior (actions) the results are valid and reliable. In the classical assumption test data is normally distributed, heteroscedasticity symptoms occur. In the hypothesis test the use of the internet has a not positive and not significant influence on political awareness