Nanobody-Based Interactomic Studies of Single Transcripts During mRNA Maturation

During and after transcription in the nucleus, messenger RNAs (mRNAs) undergo a variety of processing events before being exported to the cytoplasm through the nuclear pore complex. mRNA processing and nuclear export require a wide range of protein factors, which interact with maturing transcripts and each other to form dynamic mRNP complexes. While there are many core, essential mRNP factors, the pathways governing mRNA maturation are not uniform, and different transcripts can be associated with mRNP complexes of dramatically different composition or kinetics. To date though, it has been difficult to study RNP complexes specific to any single mRNA species, as each transcript is relatively unabundant in the cell, and few robust techniques exist to specifically purify a particular mRNP for proteomic analysis. We thus sought to develop a method to isolate mRNPs from a single transcript, allowing us to study the dynamic RNP compositions of individual mRNA maturation pathways. To optimize purifications of the protein tags required for RNP isolations, we first generated high affinity reagents targeting key tags like GFP and mCherry. Instead of traditional antibodies, we chose to use nanobodies: recombinant single domain derivatives of a heavy chain-only antibody variant found in camelids. The recombinant nature and small size of nanobodies make them ideal reagents for affinity isolations. We developed an improved pipeline for the identification of nanobody repertoires against any antigen of interest, which provided us with 25 nanobodies against GFP, the most common and robust protein tag in use. This pipeline has also allowed us to develop nanobodies against a variety of other antigens of biomedical interest. With the help of optimized reagents, we developed a two-step purification method allowing highly targeted isolations of mRNPs, starting in a budding yeast model system. In our approach, a single target transcript is tagged with MS2 hairpin sequences – these hairpins are bound specifically and with high affinity by the bacteriophage MS2 coat protein (MS2CP). In the first purification step, a chosen RNP protein known to be associated with a particular mRNA processing step of interest is Protein A-tagged and affinity isolated. From this material, anti-GFP nanobodies are used in the second step to isolate the MS2-tagged transcript of interest, through purification of MS2CP-GFP fusion proteins bound to the tag. This approach is able to efficiently and cleanly isolate a particular transcript at a chosen step of mRNP maturation. The use of an RNP factor as a separate purification target both improves overall purity and simplifies analysis by limiting heterogeneity of the mRNP mixture. Using this novel method for single mRNP isolations, we have performed a preliminary survey of transcripts with distinct sequence elements suspected to be associated with unique processing machinery. Mass spectrometric (MS) analysis of RNPs co-purified with these transcripts revealed several RNA-specific changes in composition. Most notably, introns from either a house keeping ACT1 gene or the RPS30b ribosomal protein gene led to dramatically different levels of various splicing-related proteins. These differences provide mechanistic insight into changes in the kinetics of spliceosome assembly determined by intron sequence

A Child Growth and Development Evaluation Using Weighted Product Method

The growth and development of children is one of the factors that must be considered in improving the education of a country. Because the maturity level of human resources can be maximized starting from the age of the children. According to the guidelines from the Ministry of Education of the Republic of Indonesia in 2007, there are six indicators to assess children's learning abilities. Namely moral, social, language, cognitive, motor, and also art. In this study, these indicators were used as an evaluation of the growth and development of children. The evaluation uses the weighted product (WP) method. The weighted product method is used because this method can provide a ranking of the final value of the evaluation. In addition, the WP method also has an assessment of benefits and costs as a more relevant assessment between indicators. Data collection was carried out using a questionnaire at kindergartens with an average age between 5-6 years. The results of the questionnaire will be calculated with indicators that have been given weight criteria. The results of this study were to determine students in the top five below with results between 0.65 to 0.62 with the order on behalf of Mahmud, Diko, Cindy, Denny, Riko. They will later carry out further learning actions to improve their abilities

SISTEM PAKAR IDENTIFIKASI TUNAGRAHITA DENGAN MENGGUNAKAN METODE FUZZY SUGENO

An expert system for Identification of mentally impaired using fuzzy Sugeno method mental retardation is a state of mental retardation, this condition is also known as mental retardation. Mentally retarded children are children where mental development does not take place normally, so as a result there is an inability in intellectual field, will, taste, social adjustment and so on. Identification of mental retardation is usually carried out by an expert or expert in the field of child growth and development, but actually parents can also carry out early identification of possibility mental retardation in children by observing children's behavior in everyday life, especially from how to communicate, interact socially with their peers, and the ability to imagine children. For this reason, in this study an expert system was created which can later be used as a solution to assist parents in early identification of possibility of mental retardation in children. In designing the expert system it will go through several stages, started initial stage is analyzing needs, namely by finding and collecting symptom data information and its classification obtained from an expert or an expert. The next stage is to process data information that has been obtained by building and designing a system with the proposed method using fuzzy Sugeno method. With the development of this expert system and packaged with a good display, it can greatly assist parents in predicting and recognizing whether child is mentally retarded or not. Keywords: expert system, identification, retardation, fuzzy sugeno.   ABSTRAK Sistem pakar identifikasi gangguan jiwa dengan metode fuzzy Sugeno Retardasi mental adalah suatu keadaan keterbelakangan mental, kondisi ini disebut juga dengan keterbelakangan mental. Anak tunagrahita adalah anak yang perkembangan mentalnya tidak berlangsung secara normal, sehingga mengakibatkan ketidakmampuan dalam bidang intelektual, kemauan, rasa, penyesuaian sosial dan sebagainya. Identifikasi keterbelakangan mental biasanya dilakukan oleh seorang ahli atau ahli di bidang tumbuh kembang anak, namun sebenarnya orang tua juga dapat melakukan identifikasi dini kemungkinan keterbelakangan mental pada anak dengan mengamati perilaku anak dalam kehidupan sehari-hari terutama dari cara berkomunikasi. , berinteraksi sosial dengan teman sebayanya, dan kemampuan berimajinasi anak. Untuk itu pada penelitian ini dibuat sistem pakar yang nantinya dapat digunakan sebagai solusi untuk membantu orang tua dalam mengidentifikasi secara dini kemungkinan terjadinya retardasi mental pada anak. Dalam merancang sistem pakar akan melalui beberapa tahapan, dimulai tahap awal adalah menganalisis kebutuhan yaitu dengan mencari dan mengumpulkan informasi data gejala dan klasifikasinya yang diperoleh dari seorang pakar atau pakar. Tahap selanjutnya adalah mengolah informasi data yang telah diperoleh dengan membangun dan merancang sistem dengan metode usulan menggunakan metode fuzzy Sugeno. Dengan dikembangkannya sistem pakar ini dan dikemas dengan tampilan yang baik dapat sangat membantu orang tua dalam memprediksi dan mengenali apakah anak tunagrahita atau tidak.  Kata Kunci: sistem pakar, identifikasi, tunagrahita, fuzzy sugeno

Limit points of subsequences

Let $x$ be a sequence taking values in a separable metric space and $\mathcal{I}$ be a generalized density ideal or an $F_\sigma$-ideal on the positive integers (in particular, $\mathcal{I}$ can be any Erd{\H o}s--Ulam ideal or any summable ideal). It is shown that the collection of subsequences of $x$ which preserve the set of $\mathcal{I}$-cluster points of $x$ [respectively, $\mathcal{I}$-limit points] is of second category if and only if the set of $\mathcal{I}$-cluster points of $x$ [resp., $\mathcal{I}$-limit points] coincides with the set of ordinary limit points of $x$; moreover, in this case, it is comeager. In particular, it follows that the collection of subsequences of $x$ which preserve the set of ordinary limit points of $x$ is comeager.Comment: To appear in Topology Appl. arXiv admin note: substantial text overlap with arXiv:1711.0426

PERENCANAAN PENINGKATAN KEMATANGAN TEKNOLOGI INFORMASI MENGGUNAKAN ACMM DAN TOGAF PADA POLITEKNIK XYZ

Salah satu manfaat yang dirasakan dari penggunaan teknologi informasi adalah peningkatan akurasi dan kecepatan informasi yang sangat membantu kegiatan operasional suatu lembaga atau organisasi. Politeknik XYZ sebagai salah satu organisasi / lembaga di bidang pendidikan, saat ini perlu memperhatikan peran teknologi informasi yang strategis dalam menunjang kegiatan akademik dan mengembangkan berbagai layanan akademik, khususnya bagi mahasiswa. Berdasarkan hal tersebut, dalam studi ini telah dilakukan perencanaan teknologi informasi yang sudah dipergunakan oleh Politeknik XYZ berdasarkan standar TOGAF dan melakukan analisis kematangan menggunakan ACMM., Hasil akhir dari perencanaan arsitektur yang dikembangkan dengan TOGAF dan ACMM ini dapat dijadikan acuan dalam memperbaiki peran sistem dan teknologi Informasi dalam mendukung proses bisnis organisasi di Politeknik XYZ, berupa meningkatnya awareness tentang pentingnya enterprise architecture oleh manajemen dalam tata kelola teknologi informasi dan langkah-langkah migrasi teknologi informasi

Some Tauberian theorems for Euler and Borel summability

The well-known summability methods of Euler and Borel are studied as mappings from ℓ1 into ℓ1. In this ℓ−ℓ setting, the following Tauberian results are proved: if x is a sequence that is mapped into ℓ1 by the Euler-Knopp method Er with r>0 (or the Borel matrix method) and x satisfies ∑n=0∞|xn−xn+1|n<∞, then x itself is in ℓ1

UA68/8/2 Evelyn Hooser Oral History

An interview on February 3, 1977 with Robert Penn Warren\u27s teacher Elizabeth Hooser conducted by Will Fridy

SIMULASI PENCARIAN GOLONGAN DARAH BERBASIS WEB DENGAN MENGGUNAKAN FRAMEWORK BOOTSTRAP DAN CODEIGNITER DI KABUPATAEN SIDOARJO

Website adalah suatu halaman web berisikan kumpulan informasi yang disediakan baik untuk perorangan, kelompok, atau organisasi. Sebuah webite membutuhkan sebuah server yang dapat diakses melalui jaringan online seperti internet, ataupun jaringan lokal (LAN) melalui alama yang lebih dikenal sebagai URL. Tujuan dari penelitian ini untuk mempelajari, menganalisis, dan merancang serta mengimplementasikan Simulasi Pencarian Darah Berbasis Web Menggunakan Framework Bootstrap dan Codeigniter dengan menggunakan PHP, CSS dan MySQL.Aplikasi yang dikembangkan untuk penelitian ini menggunakan Hypertext Prepocessor (PHP) yang merupakan bahasa pemrograman umum dan juga untuk pengembangan web. Selain itu juga digunakan CodeIgniter (Framework PHP), Bootstrap (Framework CSS) untuk membuat web dinamis dan responsif dengan cepat tanpa harus membuat dari awal. Hasil akhir dari penelitian ini adalah program untuk Simulasi Pencarian Darah Berbasis Web Menggunakan Framework Bootstrap dan Codeigniter yang dapat menampilkan informasi tentang profil Rumah Sakit dan PMI yang menyediakan darah dan karena ini simulasi jadi data stok darah yang tertampil pada web merupakan data buatan peneliti sendiri / tidak valid