57 research outputs found
CRISPR-Cas13a-powered electrochemical biosensors for RNA-based disease diagnostic and monitoring
No full textNucleic acids serve as specific, selective, and sensitive components in molecular diagnostics, offering efficient and high-precision results. Unlike DNA, RNA expression reflects real-time cellular activity, allowing for the monitoring of disease progression, treatment response, or environmental influences. This makes RNA a superior biomarker due to its ability to enable early disease detection, provide higher specificity, allow non-invasive sampling, and offer high sensitivity for low-abundance targets. RNA-based biosensor innovations hold significant potential for detecting genetic diseases, such as cancer, and preventing viral infections. Electrochemical biosensors have become a fast and efficient alternative to gold-standard diagnostic methods, offering simplicity, rapid response, and suitability for clinical use, including point-of-care applications. Recent advancements have integrated the CRISPR-Cas13a system with electrochemical biosensors to enhance RNA detection sensitivity and specificity. The CRISPR-Cas system, an adaptive immune mechanism in bacteria, has been widely utilized for diagnostics. Cas13a is superior to other Cas proteins for RNA detection due to its high specificity, inherent signal amplification, and ability to detect low-abundance RNA without requiring reverse transcription or amplification steps. This review summarized recent progression of CRISPR/Cas 13a and its combination with electrochemical technique, including electrochemiluminescence (ECL) and photoelectrochemical (PEC) methods. The principles and advantages of CRISPR/Cas13a, electrochemical, ECL, and PEC technique for RNA detection are described. In electrochemical-based biosensors, Cas13a recognizes and binds to the target ssRNA, triggering its trans-cleavage activity, which indiscriminately cuts nearby RNA reporters. This process alters the electrochemical signal, enabling selective and sensitive RNA detection. Finally, several examples of CRISPR/Cas13a-based electrochemical biosensors are discussed, highlighting their potential as molecular diagnostic tools for RNA detection and emphasizing their advantages in sensitivity, specificity, and rapid detection capabilities. © 2012 Published by Elsevier Ltd. Selection and/or peer-review under responsibility of Global Science and Technology Forum Pte Lt
Fatty-Acid-Binding Proteins: From Lipid Transporters to Disease Biomarkers
No full textFatty-acid-binding proteins (FABPs) serve a crucial role in the metabolism and transport of fatty acids and other hydrophobic ligands as an intracellular protein family. They are also recognized as a critical mediator in the inflammatory and ischemic pathways. FABPs are found in a wide range of tissues and organs, allowing them to contribute to various disease/injury developments that have not been widely discussed. We have collected and analyzed research journals that have investigated the role of FABPs in various diseases. Through this review, we discuss the findings on the potential of FABPs as biomarkers for various diseases in different tissues and organs, looking at their expression levels and their roles in related diseases according to available literature data. FABPs have been reported to show significantly increased expression levels in various tissues and organs associated with metabolic and inflammatory diseases. Therefore, FABPs are a promising novel biomarker that needs further development to optimize disease diagnosis and prognosis methods along with previously discovered markers
DNA biosensor for detection of Salmonella typhi from blood sample of typhoid fever patient using gold electrode modified by self-assembled monolayers of thiols
No full textIDENTIFIKASI POPULASI BAKTERI DALAM SPONS PENCUCI PIRING DENGAN METODE PCR-RFLP
No full textSpons pencuci piring 200.000 kali lebih kotor dibanding dudukan toilet. Berbagai bakteri penyebab penyakit seperti Eschericia coli, Pseudomonas dan Staphylococcus, berkembang biak di permukaan yang basah. Selain itu, 500 ribu bakteri hidup di dalam saluran pembuangan bak cuci piring. Jika spons dalam kondisi tidak kering (lembab karena direndam), maka akan menjadi markas semua bakteri. Penelitian ini bertujuan untuk menentukan populasi bakteri yang terdapat pada spons cuci piring yang basah. Identifikasi dilakukan dengan metoda molekular berbasis DNA yaitu teknik PCR-RFLP gen 16s rDNA. Metoda PCR-RFLP merupakan genetik fingerprinting yang akurat, cepat dan sederhana. Koloni bakteri yang ditumbuhkan dari spons pencuci piring di lisis dan digunakan sebagai templat untuk PCR. Hasil PCR gen 16s rDNA yang berukuran 1400 pb, dipotong dengan enzim restriksi, Hinf1. Hasil penelitian menunjukkan terdapat empat pola pemotongan yang berbeda-beda. Pola yang berbeda ini menunjukkan terdapat empat populasi yang berbeda hidup pada spons basah. Kami mengidentifikasi bahwa salah satu mikroba yang tumbuh adalah E. coli yang merupakan bakteri patogen
Mutation and Phylogenetic Analysis of Spike Glycoprotein of Indonesian Isolates of Severe-Acute-Respiratory-Syndrome-Coronavirus-2 (SARS-CoV-2)
No full textCoronavirus disease-2019 (COVID-19) is an infectious acute respiratory disease caused by SARS-CoV-2. The protein that plays a role in the entry of SARS-CoV-2 into human cells is the surface protein, or the Spike, which is thought to be the effective vaccine target to prevent SARS-CoV-2 infection. Until December 2020, Indonesia has reported 106 SARS-CoV-2 genome sequences identified from COVID-19 positive patients. The purpose of this study was to analyze the phylogenetic relationship of the Spike protein of the Indonesian isolates of SARS-CoV-2 Indonesian, as well as the virus mutations and their effects on changes in the amino acid. The 106 Indonesian SARS-CoV-2 genomes were downloaded from GISAID and the Spike nucleotide and amino acid sequences were analyzed by multiple sequence alignment (MSA) and mutation analysis using the ClustalW method. Phylogenetic trees were created using the Neighbor-Joining method in MEGA-X software. The results showed that 30 of the 106 Indonesian isolate SARS-CoV-2 Spike were 100% identical to the Wuhan-Hu-1, while the remaining 76 had experienced mutations at 1-4 sites. There were 43-point mutations in the Spike gene, 27 of which led to amino acid changes and four had not been reported in other countries. The global mutation D614G was found in 60 Indonesian isolates , of which West Java was the province with the most reports. The phylogenetic of Spike showed that the Indonesian samples have been divided into several branches that are far from Wuhan-Hu-1. This study indicates the possibility of differences in the protein structure of Indonesian isolate SARS-CoV-2 Spike that need to be further studied to manufacture a vaccine against the Indonesian strain of SARS-CoV-2
Ethyl Acetate Fraction of Moringa oleifera Leaves Induces Cell Cycle Arrest on T47D Breast Cancer Cell via G0/G1 through Cyclin D1 Expression
No full textCancer is a disease that is the leading cause of death worlwide. Exploration of a natural product containing anticancer agent provides a promising line for research on cancer. One of plant that used in traditional medicine for the cancer treatment is Moringa oleifera. The previous study has reported that the n-hexane fraction of M. oleifera leaves induce apoptosis and cell cycle arrest in T47D cells. Based on the preliminary result, ethyl acetate fraction of M. oleifera (EMO) leaves showed medium cytotoxic activity on T47D cells with IC50 values of 243.58 μg/mL and induced apoptosis cell death. This study was carried out to investigate the anticancer activity mechanism on the cellular and molecular basis of EMO against T47D breast cancer cell line by observing cell cycle progression and Cyclin D1 expression level. Analysis of the cell cycle was performed using flow cytometry and cyclin D1 expression was analyzed using the immunocytochemistry method. The result shows that EMO induced cell cycle arrest in G0/G1 phases. Immunocytochemistry assay showed that the EMO decreased expression of cyclin D1. This result indicates that EMO has a potential compound to be explored as chemo preventive agent for breast cancer
AKTIVITAS SITOTOKSIK FRAKSI ETIL ASETAT DAUN KELOR (MORINGA OLEIFERA) DAN PENGARUHNYA TERHADAP INDUKSI APOPTOSIS PADA SEL KANKER PAYUDARA T47D
No full textTanaman kelor (Moringa oleifera) adalah salah satu tanaman yang telah banyak digunakan dalam industri pengobatan tradisional. Beberapa penelitian telah melaporkan bahwa ekstrak daun M. oleifera memiliki aktivitas antiproliferatif pada beberapa sel kanker, di antaranya terhadap sel kanker hati HepG2, sel kanker paru A549, sel kanker kolon Caco-2 dan sel kanker payudara MDA-MB-231, namun penelitian terhadap sel kanker payudara T47D belum dilakukan. Oleh karena itu, penelitian ini bertujuan untuk mengetahui aktivitas antikanker daun M. oleifera terhadap sel kanker payudara T47D. Ekstrak daun M. oleifera diperoleh dengan cara ekstraksi menggunakan pelarut etanol, kemudian difraksionasi dengan pelarut n-heksana dan etil asetat. Uji sitotoksik yang dilakukan pada penelitian ini menggunakan metode MTT assay. Pengujian terhadap induksi apoptosis dilakukan dengan metode pewarnaan Annexin V-PI dan dianalisis dengan flow cytometry. Kajian molekular juga dilakukan melalui pengamatan protein regulator apoptosis, yaitu Bcl-2 dengan metode imunositokimia. Berdasarkan hasil dari MTT assay, sel T47D yang diberi perlakuan dengan fraksi etil asetat daun M. oleifera menunjukkan aktivitas medium sebagai antikanker dengan nilai IC50 sebesar 243,58 μg/mL. Pengujian terhadap apoptosis menunjukkan bahwa fraksi etil asetat daun M. oleifera dapat menginduksi apoptosis. Hasil pengamatan dengan imunositokimia menunjukkan bahwa pemberian fraksi etil asetat daun M. oleifera mampu menurunkan ekspresi protein Bcl-2.
Kata kunci: Apoptosis, Moringa oleifera, kanker payudara, sel T47
n-Hexane fraction of Moringa oleifera Lam. leaves induces apoptosis and cell cycle arrest on T47D breast cancer cell line
No full textContext: Moringa plant (Moringa oleifera) is one of the medicinal plants used as traditional medicine for the treatment of the variety of diseases including cancer. Recently, extensive research has been conducted on leaf extracts of M. oleifera to evaluate their potential cytotoxic effects. Several studies have reported anti proliferative activity of water and alcoholic extracts of leaf, bark and seed of M. oleifera on some cancer cell line, including HepG2 liver cancer cell, A549 lung cancer cell, Caco-2 colon cancer cell, MCF7 and MDA-MB-231 breast cancer cells.
Aim: To evaluate the cytotoxic effect of n-hexane fraction of M. oleifera (hMO) leaves on T47D breast cancer cells.
Methods: The M. oleifera leaves were extracted using ethanol, then fractionated with n-hexane. The cytotoxic activity was determined using MTT assay. Apoptosis and cell cycle arrest were analyzed using flow-cytometry and expression of Bcl-2 and cyclin D1 were analyzed using immunocytochemistry.
Results: Based on preliminary MTT assay, T47D treated with hMO demonstrated a medium cytotoxic effect and inhibited cell proliferation with an IC50 value of 235.58 µg/mL. Detection of apoptosis showed that hMO induced apoptosis mediated cell death in slow manner. In addition, hMO was also induce cell cycle arrest on G0-G1 and G2-M phase. Immunocytochemistry assay showed that the hMO decreased expression of anti-apoptosis protein, Bcl-2, and cell cycle regulator protein, cyclin D1, in concentration dependent manner.
Conclusions: hMO has properties to induce apoptosis and cell cycle arrest on T47D cells. hMO has a potential compound to be explored and developed as a chemo preventive agent for breast cancer. These results provide evidence for anticancer activity of M. oleifera leaves extract since it cause growth inhibition, induced apoptosis and cell cycle arrest
STUDI IN SILICO SINGLE CHAIN VARIABLE FRAGMENT (SCFV) SELEKTIF TERHADAP HORMON BASIC NATRIURETIC PEPTIDE (BNP)
No full textBasic natriuretic peptide (BNP) merupakan polipeptida yang terdiri dari 32 asam amino yang disekresikan oleh bilik jantung untuk merespon peregangan yang berlebihan pada sel otot jantung. Pengeluaran BNP dimodulasi oleh ion kalsium. BNP berpotensi untuk digunakan sebagai marker untuk meramalkan pasien yang mengalami gagal jantung. Anti BNP single chain variable fragment (Anti BNP SCFV) merupakan gabungan polipeptida antara daerah yang bervariasi pada rantai heavy (VH) dan rantai light (VL) dari immunoglobulin. Anti BNP SCFV akan berikatan dengan BNP melalui pengenalan antigen-antibodi yang biasanya berada pada daerah CDR (Complementary Determining Region) yang merupakan bagian dari SCFV. Tujuan penelitian ini adalah untuk mempelajari selektivitas docking SCFV yang diperoleh dari Protein Data Bank (PDB) dengan BNP berdasarkan parameter energi intermolekular secara in silico. SCFV terpilih dimodifikasi melalui penggantian asam amino yang berperan pada interaksi untuk mendapatkan SCFV yang lebih selektif terhadap BNP dengan energi interaksi intermolekular yang lebih rendah. Docking dilakukan menggunakan program Autodock 4.2.3. Visualisasi dilakukan menggunakan program Molegro Virtual Viewer. Hasil penelitian menunjukkan SCFV dengan ID-PDB 4OUO merupakan SCFV yang selektif terhadap BNP dengan energi intermolekular -12,81 kkal/mol, Ki 0,9712 M, namun energi ikatan yang positif: 17,33 kkal/mol. Penggantian asam amino arginin 116 menjadi histidin pada SCFV 4OUO memperlihatkan pengikatan yang lebih selektif terhadap BNP dengan energi intermolekul -13,66 kkal/mol, energi ikatan 16,47 kkal/mol, dan Ki 0,9726 M. Bagaimanapun, metode prediksi interaksi antara BNP dan SCFV perlu dikembangkan lebih lanjut untuk mendapatkan hasil yang lebih baik
Voltammetric DNA Biosensor using Gold Electrode Modified by Self Assembled Monolayer of Thiol for Detection of Mycobacterium Tuberculosis
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