Pentingnya Vaksinasi Influenza Rutin: Sebuah Pelajaran dari Data Evolusi Virus H3n2 di Indonesia Tahun 2005-2019

Pentingnya Vaksinasi Influenza Rutin: Sebuah Pelajaran dari Data Evolusi Virus H3N2 di Indonesia Tahun 2005 – 2019. Virus H3N2 menyebabkan terjadinya salah satu pandemi sejak kemunculannya pada tahun 1968 yang mengakibatkan lebih dari satu juta kematian di seluruh dunia. Hal ini disebabkan oleh adanya evolusi virus, di mana pada evolusi tersebut terjadi perubahan pada gen hemaglutinin (HA) dan neuraminidase (NA) sebagai dua glikoprotein permukaan yang berperan penting sebagai target utama dari sistem imun pejamu. Penelitian ini bertujuan untuk mengetahui pentingnya melakukan vaksinasi influenza rutin yang dibuktikan dari data evolusi virus H3N2 di Indonesia dengan melihat perubahan HA NA dan efeknya terhadap antigenisitas, di mana antigenisitas mempengaruhi kemampuan antibodi untuk mengenali virus influenza yang telah bermutasi. Terdapat 133 data HA dan 130 NA yang memenuhi kriteria inklusi dan eksklusi penelitian yang dikumpulkan dari bank data GISAID, lalu diolah dengan metode in silico dengan pembuatan pohon filogenetik menggunakan software MEGA-X dan prediksi antigenisitas menggunakan server IEDB dan Vaxijen 2.0. Pada penelitian ini terdapat evolusi pada gen HA dan NA dari virus H3N2 di Indonesia dari tahun 2005 sampai 2019 yang mengakibatkan munculnya berbagai varian epitop yang berbeda dari epitop sekuens ancestor. Di mana terdapat penurunan antigenisitas pada data gen HA dan NA tahun 2019 jika dibandingkan dengan sekuens ancestor. Penelitian ini membuktikan bahwa penting untuk melakukan vaksinasi influenza secara berkala yang dibuktikan dari adanya evolusi virus H3N2  berdasarkan adanya perubahan pada HA dan NA yang mengakibatkan adanya penurunan antigenisitas, sehingga memungkinkan virus masih terus berevolusi

Pengujian Dan Perbandingan Efektifitas Antimikroba Dari Hand Sanitizer In-House (Tahap Optimasi Kadar Etanol Terbaik untuk Membunuh Bakteri E. Coli.)

ABSTRACT Hand hygiene is an easy way to prevent the transmission of various infectious diseases. The presence of hand sanitizer has a major impact on the ease of accessing hand hygiene. Hand sanitizer is an alternative to washing hands when water and soap are not available. However, it is important to evaluate the antimicrobial effect of hand sanitizers in the market and in-house products. This study conducted a comparative research by comparing different formulations of ingredients on bactericidal effect. This study focuses on hand sanitizer and the comparative concentration of ethanol and tea tree oil to eliminate E. coli bacteria. The best alcohol concentration was 70%, compared to absolute, 80%, and 60% and there was no difference in the effectiveness of the tested hand sanitizers. Hand sanitizers manufacturing is highly dependent on active ingredients and concentrations. It is important to be thoughtfull about choosing and making hand sanitizer, so that the product can have adequate function. Hand sanitizer with optimal content and effect can reduce the incidence of infectious diseases. Keywords: Antimicrobial, Ethanol, Hand Hygine, Hand Sanitizer  ABSTRAK Hand hygine merupakan cara mudah untuk mecegah transmisi berbagai macam penyakit menular. Hadirnya hand sanitizer memberikan dampak besar dalam kemudahan akses hand hygine. Hand sanitizer menjadi alternatif mencuci tangan saat tidak tersedianya air dan sabun. Meski demikian pentingnya untuk mengevaluasi efek antimikroba yang terkandung dalam hand sanitizer yang beredar di pasar maupun produk sendiri. Studi ini melakukan penelitian komparatif dengan membandingkan formulasi bahan yang berbeda terhadap efek bakterisidal. Penelitian ini berfokus pada hand sanitizer dan tahap perbandingan kadar etanol dan konsentrasi tea tree oil untuk membunuh bakteri E. coli. Konsentrasi alkohol terbaik berdara di angka 70%, dibandingkan kengan kadar absolut, 80%, dan 60% serta tidak didapatkan perbedaan efektivitas terhadap produk hand sanitizer yang diuji. Pembuatan Hand sanitizer sangat bergantung pada bahan aktif dan konsentasi. Pentingnya untuk memperhatikan aturan dalam memilih dan membuat hand sanitizer, agar produk bisa memiliki fungsi yang adekuat. Hand sanitizer dengan kandungan dan efek yang optimal mampu menurunkan kejadian infeksi menular.Kata Kunci: Antimikroba, Etanol, Hand Hygine, Hand Sanitize

Optimasi Gelred Sebagai Pewarna Dna Dalam Biologi Molekuler

Pewarnaan DNA merupakan sebuah proses penting dalam elektroforesis gel agarosa. Proses ini memungkinkan penggunanya untuk melihat pita asam deoksiribonukleotida (DNA) hasil dari proses amplifikasi oleh teknik polymerase chain reaction (PCR) ataupun modifikasinya seperti metode restriction fragment length polymorphism (RFLP). Pewarna DNA yang umum digunakan dalam proses tersebut adalah ethidium bromide yang memiliki bahaya karena bersifat mutagenik. Pewarna DNA Gelred merupakan alternatif untuk ethidium bromide yang dinilai memiliki tingkat keamanan yang lebih baik untuk penggunanya. Penelitian ini bertujuan untuk melihat kemampuan Gelred sebagai pewarna DNA, cara terbaik untuk menggunakannya serta melihat sensitivitasnya. Pada penelitian ini 100-1000 bp DNA Ladder dengan konsentrasi berbeda digunakan sebagai penanda untuk membandingkan metode terbaik (pre-cast, post-staining dan pre-loading/pre-staining) untuk menggunakan Gelred. Selain itu dua buffer elektroforesis umum (TAE dan TBE) digunakan untuk membandingkan hasil pewarnaan DNA. Hasil dari penelitian ini didapatkan bahwa Gelred mempengaruhi migrasi DNA pada gel agarosa. Migrasi tersebut juga dipengaruhi oleh jenis buffer yang digunakan, konsentrasi agarosa, ukuran DNA, konsentrasi DNA dan metode yang digunakan (pre-cast, post-staining atau pre-loading/pre-staining). Metode terbaik untuk mendapatkan hasil visualisasi yang tajam adalah metode pre-cast dengan buffer TAE, dengan kelemahannya adalah konsentrasi dan panjang DNA sangat mempengaruhi visualisasi. Metode yang paling stabil untuk digunakan adalah metode pre-loading/pre-staining dengan buffer TAE dengan kelemahan visualisasi pita DNA yang dihasilkan tidak setajam metode pre-cast. Menurut hasil penelitian ini peneliti menyarankan penggunaan metode pre-loading/pre-staining dengan buffer TAE apabila menggunakan Gelred yang disebabkan oleh efisiensi dan kecepatan proses

Development of Amplification Refractory Mutation System PCR to Detect Androgen Receptor Gene G1773A (rs6152) Polymorphism

Androgen receptor (AR) is a ubiquitous receptor responsible for responses by androgen stimulus. Androgen, a hormone which will bind to the AR, is essentials for normal male sexual development. Nevertheless, one of the polymorphisms in the AR gene, G1733A (rs6152) have been associated with numerous clinical risks such as cardiovascular disease (CD), androgenetic alopecia, high prostate-specific antigen (PSA) levels, male infertility, recurrent spontaneous abortions and prostate cancer. This study aims to develop an alternative and cost-effective method to detect G1773A (rs6152) polymorphism. In this study, amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) using two pair of primers was used to detect the G1733A (rs6152) polymorphism. Primer design was done using http://primer1.soton.ac.uk/primer1.html online tools and then manually adjusted to increase the specificity. A total of 54 samples were screened using ARMS PCR and 2 representative samples of each allele from previous screening were used to validate the results using Sanger DNA sequencing. Among 54 subjects screened, we found 52 (96.3%) subjects carry G allele and 2 (3.7%) subjects carry A allele. No heterozygote was found in this study. The frequency of G allele was 96.97% and the frequency of A allele was 3.03%. Result validation using DNA sequencing was in agreement with ARMS-PCR method results. ARMS-PCR can be used as efficient alternative for genotyping of G1733A (rs6152) AR gene polymorphism

HBV sequences used in this study.

<p>^† Details of the GenBank Accession Numbers for all HBV sequences are provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132533#pone.0132533.s004" target="_blank">S1 Table</a>.</p><p>^‡ Five HBV/C sequences initially used in phylogenetic tree construction representing HBV/C7, C9, C10, C15, and C16 were not used in nucleotide divergence analysis because only single complete genome isolates were available for each of these subgenotypes.</p><p>^§ Core immune epitope analysis used sequences that cover the C gene region. Compared to the sequences used in the surface immune epitope analysis, only 16 HBV/C Asia sequences were used again in the core immune epitope analysis, while all S gene sequences from HBV/C Papua Pacific and the 87 HBV/C Indonesia of this study did not qualify for the core immune epitope analysis.</p><p>HBV sequences used in this study.</p

Distribution of HBsAg subtypes and HBV genotypes/subgenotypes of 271 HBV/C isolates according to their country/geographical origins in East/Southeast Asia and Papua-Pacific.

<p>^{# including 37 published complete genome sequences and 87 newly generated in this study; N. Caledonia: New Caledonia; PNG: Papua New Guinea.}</p><p>Distribution of HBsAg subtypes and HBV genotypes/subgenotypes of 271 HBV/C isolates according to their country/geographical origins in East/Southeast Asia and Papua-Pacific.</p

Phylogenetic tree of HBV/C isolates from different countries in East and Southeast Asia, and Papua-Pacific.

<p>A Bayesian phylogenetic tree analysis based on complete genome sequences showed that isolates from various subgenotypes (C1-C16) are clearly grouped into two major clusters, consistent with their geographical origins. Seven HBV/C subgenotypes (C1, C2, C5, C7, C8, C9, and C10) from East and Southeast Asia, and one (C14) from Papua (<i>light highlight</i>) were well-separated from those six subgenotypes (C6, C11, C12, C13, C15, and C16) from Papua, and from one subgenotype (C3) from Pacific, the more east region of the Papua (<i>dark highlight</i>). Although the root of subgenotype C3 phylogenetically is distanced from the subgenotypes of Papua, the isolate geographic origin, the immune epitope characteristics of surface and core proteins, and the HBsAg subtype gradient distribution showed these HBV/C3 isolates to be close to Papua subgenotypes. Therefore, the Papua and the Pacific subgenotypes are classified together into Papua-Pacific type. The diversification of the Asian type from the Papua-Pacific type started from Papua of Indonesia to the east. The other subgenotype, HBV/C4, was distanced from other subgenotypes. In this analysis, one strain (GQ358157) from Papua reported as C6 in our previous study [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132533#pone.0132533.ref023" target="_blank">23</a>] grouped into C12. We redefine this strain as a member of HBV/C12.</p

HBcAg amino acid motifs in B and T-cell epitopes of Asia and Papua-Pacific HBV/C isolates.

<p>For the sake of clarity, this figure was not drawn to scale. Among 15 amino acid positions examined within HBcAg immune epitopes of 143 isolates, we identified I/V at position c59 as the essential variation that classified HBV/C subgenotypes into two major clusters, the Asian and the Papua-Pacific (p-value <0.001; data not shown). HBV/C4 and C14 showed similar variation in most amino acids examined, with C4 and C14 having cI59 and cV59, respectively.</p

Distribution of HBV/C subtypes in the East and Southeast Asia and the Papua-Pacific.

<p>This study identified a west-to-east gradient in the distribution of HBsAg subtypes with <i>adrq+</i> (<i>red</i>) prominent in East-Southeast Asia and <i>adrq-</i> (<i>pink</i>) in the Pacific region (Vanuatu, Fiji, Tonga, and Kiribati). Interestingly, together with <i>adrq+</i>, <i>adrq-</i>indeterminate sA159/sA177 and a new pattern of <i>adrq-</i>indeterminate sV159/sV177 identified in this study were found in Papua and PNG, respectively, suggesting that the molecular admixture of HBV/C, particularly for subtype evolution, occurred in Papua and PNG with both <i>adrq-</i>indeterminate forms (<i>yellow</i>) as the transitional patterns.</p

Mean percentage nucleotide divergence of the complete genome between HBV/C subgenotypes.

<p>The total number of HBV/C isolates examined for each subgenotype is shown in bracket. Other existing subgenotypes (C7, C9, C10, C15, and C16) were not included in the genetic distance calculation since only single isolate was available for each subgenotype. Intrasubgenotype divergences are shown in bold.</p