| 1. |
- Börlin, Christoph Sebastian, 1989, et al.
(författare)
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A bioinformatic pipeline to analyze ChIP-exo datasets
- 2019
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Ingår i: Biology Methods and Protocols. - : Oxford University Press (OUP). - 2396-8923. ; 4:1
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Tidskriftsartikel (refereegranskat)abstract
- The decrease of sequencing cost in the recent years has made genome-wide studies of transcription factor (TF) binding through chromatin immunoprecipitation methods like ChIP-seq and chromatin immunoprecipitation with lambda exonuclease (ChIP-exo) more accessible to a broader group of users. Especially with ChIP-exo, it is now possible to map TF binding sites in more detail and with less noise than previously possible. These improvements came at the cost of making the analysis of the data more challenging, which is further complicated by the fact that to this date no complete pipeline is publicly available. Here we present a workflow developed specifically for ChIP-exo data and demonstrate its capabilities for data analysis. The pipeline, which is completely publicly available on GitHub, includes all necessary analytical steps to obtain a high confidence list of TF targets starting from raw sequencing reads. During the pipeline development, we emphasized the inclusion of different quality control measurements and we show how to use these so users can have confidence in their obtained results.
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| 2. |
- Goswami, Arpita, et al.
(författare)
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De novo design of anti-variant COVID-19 vaccine
- 2023
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Ingår i: Biology Methods and Protocols. - : Oxford University Press. - 2396-8923. ; 8:1
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Tidskriftsartikel (refereegranskat)abstract
- Recent studies highlight the effectiveness of hybrid Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) vaccines combining wild-type nucleocapsid and Spike proteins. We have further enhanced this strategy by incorporating delta and omicron variants' spike protein mutations. Both delta and omicron mark the shifts in viral transmissibility and severity in unvaccinated and vaccinated patients. So their mutations are highly crucial for future viral variants also. Omicron is particularly adept at immune evasion by mutating spike epitopes. The rapid adaptations of Omicron and sub-variants to spike-based vaccines and simultaneous transmissibility underline the urgency for new vaccines in the continuous battle against SARS-CoV-2. Therefore, we have added three persistent T-cell-stimulating nucleocapsid peptides similar to homologous sequences from seasonal Human Coronaviruses (HuCoV) and an envelope peptide that elicits a strong T-cell immune response. These peptides are clustered in the hybrid spike's cytoplasmic region with non-immunogenic linkers, enabling systematic arrangement. AlphaFold (Artificial intelligence-based model building) analysis suggests omitting the transmembrane domain enhances these cytoplasmic epitopes' folding efficiency which can ensure persistent immunity for CD4(+) structural epitopes. Further molecular dynamics simulations validate the compact conformation of the modeled structures and a flexible C-terminus region. Overall, the structures show stability and less conformational fluctuation throughout the simulation. Also, the AlphaFold predicted structural epitopes maintained their folds during simulation to ensure the specificity of CD4(+) T-cell response after vaccination. Our proposed approach may provide options for incorporating diverse anti-viral T-cell peptides, similar to HuCoV, into linker regions. This versatility can be promising to address outbreaks and challenges posed by various viruses for effective management in this era of innovative vaccines.
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| 3. |
- López Clinton, Samantha, 1994-
(författare)
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Microcosmos explorers : foldscope workshop for science outreach in Mexican schools
- 2023
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Ingår i: Biology Methods and Protocols. - 2396-8923. ; 8:1
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Tidskriftsartikel (refereegranskat)abstract
- Foldscopes are ultra-low-cost paper microscopes invented by Manu Prakash and Jim Cybulski at Stanford University. They are about as light as a pencil and waterproof, all whilst offering similar optic quality to traditional microscopes. Foldscopes do not require electricity or glass slides to be used, which increases the possibilities of their use in education and outreach activities with children or people with disabilities. In 2019, thanks to a material grant of 100 foldscopes from One World Science and additional purchased foldscopes, I designed and implemented a science workshop called Exploradores del Microcosmos, or Explorers of Microcosmos in English. The aim of the workshop was to help make microscopy more accessible, in particular at underfunded schools, and stimulate active learning about ecosystems and evolution in the participants. Within this article, I describe the workshop and relay my personal insights and reflections on its execution across multiple schools and groups in Mexico.
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| 4. |
- Siddiqui, Amna Jabbar, et al.
(författare)
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Serum metallomics reveals insights into the associations of elements with the progression of preleukemic diseases toward acute leukemia
- 2024
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Ingår i: Biology Methods and Protocols. - : Oxford University Press. - 2396-8923. ; 9:1
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Tidskriftsartikel (refereegranskat)abstract
- Acute leukemia (AL) is a critical neoplasm of white blood cells with two main subtypes: acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). This study is focused on understanding the association of the preleukemic disease aplastic anemia (APA) with ALL and AML at metallomic level, using healthy subjects as a control. In this study, a validated and efficient inductively coupled plasma-mass spectrometry/MS-based workflow was employed to profile a total of 13 metallomic features. The study encompassed 41 patients with AML, 62 patients with ALL, 46 patients with APA, and 55 age-matched healthy controls. The metallomic features consisted of eight essential elements (Ca, Co, Cu, Fe, Mg, Mn, Se, and Zn) and five non-essential/toxic elements (Ag, Cd, Cr, Ni, and Pb). Six out of the 13 elements were found to be substantially different (P < .05) using absolute concentrations between serum samples of AL (ALL and AML) and preleukemia (APA) patients in comparison with healthy subjects. Elements including magnesium, calcium, iron, copper, and zinc were upregulated and only one element (chromium) was downregulated in serum samples of disease when compared with healthy subjects. Through the utilization of both univariate tests and multivariate classification modeling, it was determined that chromium exhibited a progressive behavior among the studied elements. Specifically, chromium displayed a sequential upregulation from healthy individuals to preleukemic disease (APA), and ultimately in patients diagnosed with ALL. Overall, metallomic-based biomarkers may have the utility to predict the association of APA with ALL.
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| 5. |
- Vihinen, Mauno
(författare)
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Guidelines for systematic reporting of sequence alignments
- 2020
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Ingår i: Biology Methods and Protocols. - : Oxford University Press (OUP). - 2396-8923. ; 5:1
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Tidskriftsartikel (refereegranskat)abstract
- Bioinformatics methods are increasingly needed and used to analyze and interpret extensive datasets many of which are produced by diverse high-throughput technologies. Unfortunately, it is quite common that published articles do not contain sufficient information to allow the reader to fully comprehend and repeat computational and other studies. Guidelines were developed for reporting studies and results from sequence alignment. Brief and concise checklist of required data items was compiled making it easy to provide necessary details. Implementation of the guidelines requires similar meticulous attitude toward details as other parts of publications. If the journal does not allow reporting full details in the main article, it can be provided in supplementary material. It is important to make the alignments available. Systematic and detailed description of bioinformatics analyses adds to the value of papers and makes it easier for the scientific community to evaluate, understand, verify, and extend the published articles and their results.
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| 6. |
- Wang, Liya
(författare)
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Quantitative solid-phase assay to measure deoxynucleoside triphosphate pools
- 2018
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Ingår i: Biology methods & protocols. - : Oxford University Press (OUP). - 2396-8923. ; 3
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Tidskriftsartikel (refereegranskat)abstract
- deoxynucleoside triphosphate (dNTPs) are the reduced nucleotides used as the building blocks and energy source for deoxyribonucleic acid (DNA) replication and maintenance in all living systems. They are present in highly regulated amounts and ratios in the cell, and their balance has been implicated in the most important cell processes, from determining the fidelity of DNA replication to affecting cell fate. Furthermore, many cancer drugs target biosynthetic enzymes in dNTP metabolism, and mutations in genes directly or indirectly affecting these pathways that are the cause of devastating diseases. The accurate and systematic measurement of these pools is key to understanding the mechanisms behind these diseases and their treatment. We present a new method for measuring dNTP pools from biological samples, utilizing the current state-of-the-art polymerase method, modified to a solid-phase setting and optimized for larger scale measurements.
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