Biology Methods and Protocols最新文献

{"title":"Advancing age grading techniques for <i>Glossina morsitans morsitans</i>, vectors of African trypanosomiasis, through mid-infrared spectroscopy and machine learning.","authors":"Mauro Pazmiño-Betancourth, Ivan Casas Gómez-Uribarri, Karina Mondragon-Shem, Simon A Babayan, Francesco Baldini, Lee Rafuse Haines","doi":"10.1093/biomethods/bpae058","DOIUrl":"10.1093/biomethods/bpae058","url":null,"abstract":"<p><p>Tsetse are the insects responsible for transmitting African trypanosomes, which cause sleeping sickness in humans and animal trypanosomiasis in wildlife and livestock. Knowing the age of these flies is important when assessing the effectiveness of vector control programs and modelling disease risk. Current methods to assess fly age are, however, labour-intensive, slow, and often inaccurate as skilled personnel are in short supply. Mid-infrared spectroscopy (MIRS), a fast and cost-effective tool to accurately estimate several biological traits of insects, offers a promising alternative. This is achieved by characterising the biochemical composition of the insect cuticle using infrared light coupled with machine-learning (ML) algorithms to estimate the traits of interest. We tested the performance of MIRS in estimating tsetse sex and age for the first-time using spectra obtained from their cuticle. We used 541 insectary-reared <i>Glossina m. morsitans</i> of two different age groups for males (5 and 7 weeks) and three age groups for females (3 days, 5 weeks, and 7 weeks). Spectra were collected from the head, thorax, and abdomen of each sample. ML models differentiated between male and female flies with a 96% accuracy and predicted the age group with 94% and 87% accuracy for males and females, respectively. The key infrared regions important for discriminating sex and age classification were characteristic of lipid and protein content. Our results support the use of MIRS as a rapid and accurate way to identify tsetse sex and age with minimal pre-processing. Further validation using wild-caught tsetse could pave the way for this technique to be implemented as a routine surveillance tool in vector control programmes.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae058"},"PeriodicalIF":2.5,"publicationDate":"2024-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11407438/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142297453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NanoMGT: Marker gene typing of low complexity mono-species metagenomic samples using noisy long reads.","authors":"Malte B Hallgren, Philip T L C Clausen, Frank M Aarestrup","doi":"10.1093/biomethods/bpae057","DOIUrl":"https://doi.org/10.1093/biomethods/bpae057","url":null,"abstract":"<p><p>Rapid advancements in sequencing technologies have led to significant progress in microbial genomics, yet challenges persist in accurately identifying microbial strain diversity in metagenomic samples, especially when working with noisy long-read data from platforms like Oxford Nanopore Technologies (ONT). In this article, we introduce NanoMGT, a tool designed to enhance marker gene typing in low-complexity mono-species samples, leveraging the unique properties of long reads. NanoMGT excels in its ability to accurately identify mutations amidst high error rates, ensuring the reliable detection of multiple strain-specific marker genes. Our tool implements a novel scoring system that rewards mutations co-occurring across different reads and penalizes densely grouped, likely erroneous variants, thereby achieving a good balance between sensitivity and precision. A comparative evaluation of NanoMGT, using a simulated multi-strain sample of seven bacterial species, demonstrated superior performance relative to existing tools and the advantages of using a threshold-based filtering approach to calling minority variants in ONT's sequencing data. NanoMGT's potential as a post-binning tool in metagenomic pipelines is particularly notable, enabling researchers to more accurately determine specific alleles and understand strain diversity in microbial communities. Our findings have significant implications for clinical diagnostics, environmental microbiology, and the broader field of genomics. The findings offer a reliable and efficient approach to marker gene typing in complex metagenomic samples.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae057"},"PeriodicalIF":2.5,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11387619/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142297456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Automatic detection of fish scale circuli using deep learning.","authors":"Nora N Hanson, James P Ounsley, Jason Henry, Kasim Terzić, Bruno Caneco","doi":"10.1093/biomethods/bpae056","DOIUrl":"10.1093/biomethods/bpae056","url":null,"abstract":"<p><p>Teleost fish scales form distinct growth rings deposited in proportion to somatic growth in length, and are routinely used in fish ageing and growth analyses. Extraction of incremental growth data from scales is labour intensive. We present a fully automated method to retrieve this data from fish scale images using Convolutional Neural Networks (CNNs). Our pipeline of two CNNs automatically detects the centre of the scale and individual growth rings (circuli) along multiple radial transect emanating from the centre. The focus detector was trained on 725 scale images and achieved an average precision of 99%; the circuli detector was trained on 40 678 circuli annotations and achieved an average precision of 95.1%. Circuli detections were made with less confidence in the freshwater zone of the scale image where the growth bands are most narrowly spaced. However, the performance of the circuli detector was similar to that of another human labeller, highlighting the inherent ambiguity of the labelling process. The system predicts the location of scale growth rings rapidly and with high accuracy, enabling the calculation of spacings and thereby growth inferences from salmon scales. The success of our method suggests its potential for expansion to other species.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae056"},"PeriodicalIF":2.5,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11330318/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142000884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harmonizing immune cell sequences for computational analysis with large language models.","authors":"Areej Alsaafin, Hamid R Tizhoosh","doi":"10.1093/biomethods/bpae055","DOIUrl":"https://doi.org/10.1093/biomethods/bpae055","url":null,"abstract":"<p><p>We present SEQuence Weighted Alignment for Sorting and Harmonization (Seqwash), an algorithm designed to process sequencing profiles utilizing large language models. Seqwash <i>harmonizes</i> immune cell sequences into a unified representation, empowering LLMs to embed meaningful patterns while eliminating irrelevant information. Evaluations using immune cell sequencing data showcase Seqwash's efficacy in standardizing profiles, leading to improved feature quality and enhanced performance in both supervised and unsupervised downstream tasks for sequencing data.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae055"},"PeriodicalIF":2.5,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11407694/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142297455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simple protocol for combined extraction of exocrine secretions and RNA in small arthropods.","authors":"David Fröhlich, Michaela Bodner, Günther Raspotnig, Christoph Hahn","doi":"10.1093/biomethods/bpae054","DOIUrl":"10.1093/biomethods/bpae054","url":null,"abstract":"<p><p>The integration of data from multiple sources and analytical techniques to obtain novel insights and answer challenging questions is a hallmark of modern science. In arthropods, exocrine secretions may act as pheromones, defensive substances, antibiotics, as well as surface protectants, and as such they play a crucial role in ecology and evolution. Exocrine chemical compounds are frequently characterized by gas chromatography-mass spectrometry. Technological advances of recent years now allow us to routinely characterize the total gene complement transcribed in a particular biological tissue, often in the context of experimental treatment, via RNAseq. We here introduce a novel methodological approach to successfully characterize exocrine secretions <i>and</i> full transcriptomes of one and the same individual of oribatid mites. We found that chemical extraction prior to RNA extraction had only minor effects on the total RNA integrity. De novo transcriptomes obtained from such combined extractions were of comparable quality to those assembled for samples that were subject to RNA extraction only, indicating that combined chemical/RNA extraction is perfectly suitable for phylotranscriptomic studies. However, in-depth analysis of RNA expression analysis indicates that chemical extraction prior to RNAseq may affect transcript degradation rates, similar to the effects reported in previous studies comparing RNA extraction protocols. With this pilot study, we demonstrate that profiling chemical secretions and RNA expression levels from the same individual is methodologically feasible, paving the way for future research to understand the genes and pathways underlying the syntheses of biogenic chemical compounds. Our approach should be applicable broadly to most arachnids, insects, and other arthropods.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae054"},"PeriodicalIF":2.5,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11316613/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141917564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deep learning pipeline reveals key moments in human embryonic development predictive of live birth after in vitro fertilization.","authors":"Camilla Mapstone, Helen Hunter, Daniel Brison, Julia Handl, Berenika Plusa","doi":"10.1093/biomethods/bpae052","DOIUrl":"10.1093/biomethods/bpae052","url":null,"abstract":"<p><p>Demand for in vitro fertilization (IVF) treatment is growing; however, success rates remain low partly due to difficulty in selecting the best embryo to be transferred. Current manual assessments are subjective and may not take advantage of the most informative moments in embryo development. Here, we apply convolutional neural networks (CNNs) to identify key windows in pre-implantation human development that can be linked to embryo viability and are therefore suitable for the early grading of IVF embryos. We show how machine learning models trained at these developmental time points can be used to refine overall embryo viability assessment. Exploiting the well-known capabilities of transfer learning, we illustrate the performance of CNN models for very limited datasets, paving the way for the use on a clinic-by-clinic basis, catering for local data heterogeneity.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae052"},"PeriodicalIF":2.5,"publicationDate":"2024-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11305813/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141903164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The landscape of RNA 3D structure modeling with transformer networks.","authors":"Sumit Tarafder, Rahmatullah Roche, Debswapna Bhattacharya","doi":"10.1093/biomethods/bpae047","DOIUrl":"10.1093/biomethods/bpae047","url":null,"abstract":"<p><p>Transformers are a powerful subclass of neural networks catalyzing the development of a growing number of computational methods for RNA structure modeling. Here, we conduct an objective and empirical study of the predictive modeling accuracy of the emerging transformer-based methods for RNA structure prediction. Our study reveals multi-faceted complementarity between the methods and underscores some key aspects that affect the prediction accuracy.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae047"},"PeriodicalIF":2.5,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11244692/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141617300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine learning of cellular metabolic rewiring.","authors":"Joao B Xavier","doi":"10.1093/biomethods/bpae048","DOIUrl":"10.1093/biomethods/bpae048","url":null,"abstract":"<p><p>Metabolic rewiring allows cells to adapt their metabolism in response to evolving environmental conditions. Traditional metabolomics techniques, whether targeted or untargeted, often struggle to interpret these adaptive shifts. Here, we introduce <i>MetaboLiteLearner</i>, a lightweight machine learning framework that harnesses the detailed fragmentation patterns from electron ionization (EI) collected in scan mode during gas chromatography/mass spectrometry to predict changes in the metabolite composition of metabolically adapted cells. When tested on breast cancer cells with different preferences to metastasize to specific organs, <i>MetaboLiteLearner</i> predicted the impact of metabolic rewiring on metabolites withheld from the training dataset using only the EI spectra, without metabolite identification or pre-existing knowledge of metabolic networks. Despite its simplicity, the model learned captured shared and unique metabolomic shifts between brain- and lung-homing metastatic lineages, suggesting cellular adaptations associated with metastasis to specific organs. Integrating machine learning and metabolomics paves the way for new insights into complex cellular adaptations.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae048"},"PeriodicalIF":2.5,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11249387/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141621138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-throughput evaluation of hemolytic activity through precise measurement of colony and hemolytic zone sizes of engineered <i>Bacillus subtilis</i> on blood agar.","authors":"Takahiro Bamba, Rina Aoki, Yoshimi Hori, Shu Ishikawa, Ken-Ichi Yoshida, Naoaki Taoka, Shingo Kobayashi, Hisashi Yasueda, Akihiko Kondo, Tomohisa Hasunuma","doi":"10.1093/biomethods/bpae044","DOIUrl":"10.1093/biomethods/bpae044","url":null,"abstract":"<p><p>Biosurfactants have remarkable characteristics, such as environmental friendliness, high safety, and excellent biodegradability. Surfactin is one of the best-known biosurfactants produced by <i>Bacillus subtilis</i>. Because the biosynthetic pathways of biosurfactants, such as surfactin, are complex, mutagenesis is a useful alternative to typical metabolic engineering approaches for developing high-yield strains. Therefore, there is a need for high-throughput and accurate screening methods for high-yield strains derived from mutant libraries. The blood agar lysis method, which takes advantage of the hemolytic activity of biosurfactants, is one way of determining their concentration. This method includes inoculating microbial cells onto blood-containing agar plates, and biosurfactant production is assessed based on the size of the hemolytic zone formed around each colony. Challenges with the blood agar lysis method include low experimental reproducibility and a lack of established protocols for high-throughput screening. Therefore, in this study, we investigated the effects of the inoculation procedure and media composition on the formation of hemolytic zones. We also developed a workflow to evaluate the number of colonies using robotics. The results revealed that by arranging colonies at appropriate intervals and measuring the areas of colonies and hemolytic rings using image analysis software, it was possible to accurately compare the hemolytic activity among several colonies. Although the use of the blood agar lysis method for screening is limited to surfactants exhibiting hemolytic activity, it is believed that by considering the insights gained from this study, it can contribute to the accurate screening of strains with high productivity.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae044"},"PeriodicalIF":2.5,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11219306/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141499181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"KCL TEST: an open-source inspired asymptomatic SARS-CoV-2 surveillance programme in an academic institution.","authors":"Joana Reis de Andrade, Edward Scourfield, Shilpa Lekhraj Peswani-Sajnani, Kate Poulton, Thomas Ap Rees, Paniz Khooshemehri, George Doherty, Stephanie Ong, Iustina-Francisca Ivan, Negin Goudarzi, Isaac Gardiner, Estelle Caine, Thomas J A Maguire, Daniel Leightley, Luis Torrico, Alex Gasulla, Angel Menendez-Vazquez, Ana Maria Ortega-Prieto, Suzanne Pickering, Jose M Jimenez-Guardeño, Rahul Batra, Sona Rubinchik, Aaron V F Tan, Amy Griffin, David Sherrin, Stelios Papaioannou, Celine Trouillet, Hannah E Mischo, Victoriano Giralt, Samantha Wilson, Martin Kirk, Stuart J D Neil, Rui Pedro Galao, Jo Martindale, Charles Curtis, Mark Zuckerman, Reza Razavi, Michael H Malim, Rocio T Martinez-Nunez","doi":"10.1093/biomethods/bpae046","DOIUrl":"10.1093/biomethods/bpae046","url":null,"abstract":"<p><p>Rapid and accessible testing was paramount in the management of the COVID-19 pandemic. Our university established KCL TEST: a SARS-CoV-2 asymptomatic testing programme that enabled sensitive and accessible PCR testing of SARS-CoV-2 RNA in saliva. Here, we describe our learnings and provide our blueprint for launching diagnostic laboratories, particularly in low-resource settings. Between December 2020 and July 2022, we performed 158277 PCRs for our staff, students, and their household contacts, free of charge. Our average turnaround time was 16 h and 37 min from user registration to result delivery. KCL TEST combined open-source automation and in-house non-commercial reagents, which allows for rapid implementation and repurposing. Importantly, our data parallel those of the UK Office for National Statistics, though we detected a lower positive rate and virtually no delta wave. Our observations strongly support regular asymptomatic community testing as an important measure for decreasing outbreaks and providing safe working spaces. Universities can therefore provide agile, resilient, and accurate testing that reflects the infection rate and trend of the general population. Our findings call for the early integration of academic institutions in pandemic preparedness, with capabilities to rapidly deploy highly skilled staff, as well as develop, test, and accommodate efficient low-cost pipelines.</p>","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":"9 1","pages":"bpae046"},"PeriodicalIF":2.5,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11238426/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141591614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}