Diana Frimpong, Angela C Shore, Ben Gardner, Claire Newton, Joya Pawade, Jonathan Frost, Laura Atherton, Nick Stone
During post chemotherapy surgery for ovarian cancer, it is important to ensure that any residual disease is carefully assessed and removed. The assessment remains subjective, despite clear evidence of the benefits of complete macroscopic resection. In this work, we have considered Raman spectroscopy as a possible tool for residual disease assessment by exploring its ability to correctly classify ovarian cancer from benign and borderline tissues. Samples from seventy-three participants were analysed (n=20 benign, n=11 borderline and n=42 cancer) using a multivariate analysis model. All models shown utilised validation with leave one participant out cross-validation. In ovarian tissue this model achieved 94% sensitivity and 98% specificity for prediction of cancer from benign and 98% sensitivity and 89% specificity for prediction of cancer from borderline. Thorough assessment of the surrounding peritoneal tissues is extremely important. For these peritoneal tissues taken from participants with advanced ovarian cancer, the model achieved 78% sensitivity and 84% specificity for prediction of cancerous peritoneum from benign peritoneum in participants who had primary surgery and 68% sensitivity and 81% specificity in participants who had post chemotherapy surgery. This demonstrates viability of Raman spectroscopy for assessment of ovarian cancer.
{"title":"Raman spectroscopy of ovarian and peritoneal tissue in the assessment of ovarian cancer","authors":"Diana Frimpong, Angela C Shore, Ben Gardner, Claire Newton, Joya Pawade, Jonathan Frost, Laura Atherton, Nick Stone","doi":"10.1039/d4an01293c","DOIUrl":"https://doi.org/10.1039/d4an01293c","url":null,"abstract":"During post chemotherapy surgery for ovarian cancer, it is important to ensure that any residual disease is carefully assessed and removed. The assessment remains subjective, despite clear evidence of the benefits of complete macroscopic resection. In this work, we have considered Raman spectroscopy as a possible tool for residual disease assessment by exploring its ability to correctly classify ovarian cancer from benign and borderline tissues. Samples from seventy-three participants were analysed (n=20 benign, n=11 borderline and n=42 cancer) using a multivariate analysis model. All models shown utilised validation with leave one participant out cross-validation. In ovarian tissue this model achieved 94% sensitivity and 98% specificity for prediction of cancer from benign and 98% sensitivity and 89% specificity for prediction of cancer from borderline. Thorough assessment of the surrounding peritoneal tissues is extremely important. For these peritoneal tissues taken from participants with advanced ovarian cancer, the model achieved 78% sensitivity and 84% specificity for prediction of cancerous peritoneum from benign peritoneum in participants who had primary surgery and 68% sensitivity and 81% specificity in participants who had post chemotherapy surgery. This demonstrates viability of Raman spectroscopy for assessment of ovarian cancer.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"85 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Elie SARKEES, Vincent Vuiblet, Fayek Taha, Olivier Piot
The global rise of end-stage renal disease is leading to an increase in kidney transplants. Transplants survival are accompanied by more inflammation and rejection cases. Traditional laboratory analyses often lack accuracy, and graft biopsies - the current gold standard - are considered invasive and risky means. This highlights an unmet need for innovative diagnostic and monitoring methods of graft rejection and inflammation. This study explores the potential of Fourier-Transform Infrared spectroscopy of fresh urine for diagnosing kidney transplant inflammation. Urine samples were collected from kidney transplant patients who are under regular surveillance. An unsupervised method of spectral data analysis, especially Uniform Manifold Approximation and Projection (UMAP), was initially employed. However, it was unable to reveal a clear distinction between control and pathological conditions. Subsequently, two machine learning models - SVM and Gradient Boosting - were employed to categorise participants into pathologic or control groups; achieving a diagnostic accuracy of 77.78%. The study also evaluated other factors that could affect model performance, including urine biochemical composition, type of inflammation, and patient's medication history. The inherent variability of the urine, attributed to factors such as diet and medications, poses challenges to identifying robust spectroscopic markers. Nevertheless, mid-infrared spectroscopy offers a promising, non-invasive approach for diagnosing kidney transplant disorders. Further research is essential to provide more advanced prediction models and meet the criteria for potential clinical deployment.
{"title":"Exploring the Potential of Fourier Transform-infrared Spectroscopy of Urine for Non-invasive Monitoring of Inflammation associated with Kidney Transplant","authors":"Elie SARKEES, Vincent Vuiblet, Fayek Taha, Olivier Piot","doi":"10.1039/d4an01459f","DOIUrl":"https://doi.org/10.1039/d4an01459f","url":null,"abstract":"The global rise of end-stage renal disease is leading to an increase in kidney transplants. Transplants survival are accompanied by more inflammation and rejection cases. Traditional laboratory analyses often lack accuracy, and graft biopsies - the current gold standard - are considered invasive and risky means. This highlights an unmet need for innovative diagnostic and monitoring methods of graft rejection and inflammation. This study explores the potential of Fourier-Transform Infrared spectroscopy of fresh urine for diagnosing kidney transplant inflammation. Urine samples were collected from kidney transplant patients who are under regular surveillance. An unsupervised method of spectral data analysis, especially Uniform Manifold Approximation and Projection (UMAP), was initially employed. However, it was unable to reveal a clear distinction between control and pathological conditions. Subsequently, two machine learning models - SVM and Gradient Boosting - were employed to categorise participants into pathologic or control groups; achieving a diagnostic accuracy of 77.78%. The study also evaluated other factors that could affect model performance, including urine biochemical composition, type of inflammation, and patient's medication history. The inherent variability of the urine, attributed to factors such as diet and medications, poses challenges to identifying robust spectroscopic markers. Nevertheless, mid-infrared spectroscopy offers a promising, non-invasive approach for diagnosing kidney transplant disorders. Further research is essential to provide more advanced prediction models and meet the criteria for potential clinical deployment.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"28 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143518288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yan Ge, Xingxing Huang, Baojian Zhang, Zhixiong Song, Xusheng Tang, Shuai Shao, Lujiale Guo, Peng Liang, Bei Li
Viscosity is an important characteristic of fluids. Microfluidics has shown significant advantages in the viscosity measurement of biopharmaceuticals, especially in meeting the needs of low sample volumes and accurately controlling microscale fluids. However, the viscosity chip of the traditional straight channel structure has limitations, and the processing technology is also facing challenges. In this study, a variable cross-section microfluidic chip structure was designed and successfully manufactured by photocuring 3D printing technology. A digital-printed (DP) microfluidic viscometer was realized by a pressure-driven flow combined with optical imaging. The device measures the change in sample viscosity with shear rate by recording the change in pressure and flow velocity with time. The whole experiment requires only 25 μl of reagents per time, and the single experiment time is less than 2 minutes, which not only reduces the consumption of samples dramatically, but also improves the efficiency of the experiment significantly. Compared with commercial viscometers, our measurements are accurate and capable of supporting non-Newtonian fluids. The proposed platform provides good cost-effectiveness and operational simplicity and lays the methodological foundation for viscosity measurements of more complex properties of fluids.
{"title":"Measurement of fluid viscosity based on pressure-driven flow digital-printed microfluidics","authors":"Yan Ge, Xingxing Huang, Baojian Zhang, Zhixiong Song, Xusheng Tang, Shuai Shao, Lujiale Guo, Peng Liang, Bei Li","doi":"10.1039/d4an01550a","DOIUrl":"https://doi.org/10.1039/d4an01550a","url":null,"abstract":"Viscosity is an important characteristic of fluids. Microfluidics has shown significant advantages in the viscosity measurement of biopharmaceuticals, especially in meeting the needs of low sample volumes and accurately controlling microscale fluids. However, the viscosity chip of the traditional straight channel structure has limitations, and the processing technology is also facing challenges. In this study, a variable cross-section microfluidic chip structure was designed and successfully manufactured by photocuring 3D printing technology. A digital-printed (DP) microfluidic viscometer was realized by a pressure-driven flow combined with optical imaging. The device measures the change in sample viscosity with shear rate by recording the change in pressure and flow velocity with time. The whole experiment requires only 25 μl of reagents per time, and the single experiment time is less than 2 minutes, which not only reduces the consumption of samples dramatically, but also improves the efficiency of the experiment significantly. Compared with commercial viscometers, our measurements are accurate and capable of supporting non-Newtonian fluids. The proposed platform provides good cost-effectiveness and operational simplicity and lays the methodological foundation for viscosity measurements of more complex properties of fluids.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"164 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Timely and accurate diagnosis of RNA viruses, represented by the SARS-CoV-2, is the foundation for protecting people from health threats. Currently, direct detection of viral RNA genes remains the most accurate detection method. Herein, a rapid, ultrasensitive, no heating equipment required and quantitative detection CRISPR/Cas13a based evanescent wave fluorescence biosensing platform for detection of SARS-CoV-2 gene is reported. The collateral effect of CRISPR/Cas13a for RNA is combined with a self-driven enzyme-free hybridization chain reaction (HCR) as a signal amplification step. When the initiator RNA strand is cleaved by Cas13a, the downstream signal amplification induced by HCR will be blocked, and a multiple crRNA strategy is used to enhance cleavage efficiency. The newly designed HCR assemblies will be captured by the cDNA modified optical fiber and generate a higher-intensity fluorescence signal induced by the evanescent field. The CRISPR/Cas13a-HCR evanescent wave fluorescence biosensing platform is capable of detection of SARS-CoV-2 with LOD of 0.47 copies/μL and the detection time is within 35 min. The spike recovery tests in saliva and high specific capability have demonstrated the potential of this method for point-of-care diagnosis. This method is also suitable for the detection of other RNA viruses, without the need to alter any design of the HCR component, only the corresponding crRNA needs to be replaced.
{"title":"A CRISPR/Cas13a-based and hybridization chain reaction coupled evanescent wave biosensor for SARS-CoV-2 gene detection","authors":"Yang Li, Yikan Zhao, Zhihao Yi, Shitong Han","doi":"10.1039/d4an01584c","DOIUrl":"https://doi.org/10.1039/d4an01584c","url":null,"abstract":"Timely and accurate diagnosis of RNA viruses, represented by the SARS-CoV-2, is the foundation for protecting people from health threats. Currently, direct detection of viral RNA genes remains the most accurate detection method. Herein, a rapid, ultrasensitive, no heating equipment required and quantitative detection CRISPR/Cas13a based evanescent wave fluorescence biosensing platform for detection of SARS-CoV-2 gene is reported. The collateral effect of CRISPR/Cas13a for RNA is combined with a self-driven enzyme-free hybridization chain reaction (HCR) as a signal amplification step. When the initiator RNA strand is cleaved by Cas13a, the downstream signal amplification induced by HCR will be blocked, and a multiple crRNA strategy is used to enhance cleavage efficiency. The newly designed HCR assemblies will be captured by the cDNA modified optical fiber and generate a higher-intensity fluorescence signal induced by the evanescent field. The CRISPR/Cas13a-HCR evanescent wave fluorescence biosensing platform is capable of detection of SARS-CoV-2 with LOD of 0.47 copies/μL and the detection time is within 35 min. The spike recovery tests in saliva and high specific capability have demonstrated the potential of this method for point-of-care diagnosis. This method is also suitable for the detection of other RNA viruses, without the need to alter any design of the HCR component, only the corresponding crRNA needs to be replaced.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"12 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ajay Kumar Sharma, Poonam Sharma, Pushkar Mehara, Pralay Das
In this study, the application of alkanolamines as an optical CO2-responsive medium in combination with galvinol (GALH) as a reversible quantitative colorimetric sensor is proposed. Monoethanolamine (MEA) and diethanolamine (DEA), as the simplest primary (1°) and secondary (2°) alkanolamines, were explored for the colorimetric CO2 response attributed to their well-known CO2 absorption ability on an industrial scale from more than six decades. Further, the anionic form of GALH (GAL−) generates an intense purple color that allows its application for colorimetric measurement which has been very less explored for optical sensing purposes. A reversible, convenient, and simplified colorimetric detection of CO2 gas was performed using GALH in combination with MEA and DEA in CH3CN solvent, which showed limits of detection (LODs) for CO2 gas as low as ∼19 and ∼31 ppm respectively with recyclabilities. The main feature of the developed CO2 sensor is its non-aqueous medium application, which makes it ideal for producing error-free results under different humidity conditions.
{"title":"Molecular combination between alkanolamines and galvinol for ratiometric colorimetric sensing of CO2 gas","authors":"Ajay Kumar Sharma, Poonam Sharma, Pushkar Mehara, Pralay Das","doi":"10.1039/d4an01557f","DOIUrl":"https://doi.org/10.1039/d4an01557f","url":null,"abstract":"In this study, the application of alkanolamines as an optical CO<small><sub>2</sub></small>-responsive medium in combination with galvinol (GALH) as a reversible quantitative colorimetric sensor is proposed. Monoethanolamine (MEA) and diethanolamine (DEA), as the simplest primary (1°) and secondary (2°) alkanolamines, were explored for the colorimetric CO<small><sub>2</sub></small> response attributed to their well-known CO<small><sub>2</sub></small> absorption ability on an industrial scale from more than six decades. Further, the anionic form of GALH (GAL<small><sup>−</sup></small>) generates an intense purple color that allows its application for colorimetric measurement which has been very less explored for optical sensing purposes. A reversible, convenient, and simplified colorimetric detection of CO<small><sub>2</sub></small> gas was performed using GALH in combination with MEA and DEA in CH<small><sub>3</sub></small>CN solvent, which showed limits of detection (LODs) for CO<small><sub>2</sub></small> gas as low as ∼19 and ∼31 ppm respectively with recyclabilities. The main feature of the developed CO<small><sub>2</sub></small> sensor is its non-aqueous medium application, which makes it ideal for producing error-free results under different humidity conditions.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"28 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143496163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The overuse and misuse of antibiotics have caused the development of antimicrobial resistance (AMR), which poses a significant threat to human health. Antimicrobial susceptibility testing (AST) serves as an effective tool for assessing the susceptibility of pathogens infecting patients and guiding the precise use of antibiotics. The conventional AST method, however, is limited by prolonged incubation times and high reagent consumption. In this study, we introduce an integrated microfluidic platform, enabling multiple AST and minimum inhibitory concentration (MIC) determinations after 2 hours incubation. Sample loading is achieved using a self-priming and vacuum-driven approach, enhancing operational feasibility and preventing cross-contamination during reagent pre-coating. Moreover, the use of chips with pre-coated antibiotics minimizes the need for reagent handling off-chip, thereby enhancing the flexibility of the microfluidic device and making the platform easy-to-use. The AST on-chip results for Klebsiella pneumoniae (K. pneumoniae) S1 correlate well with broth dilution methods. This integrated microfluidic platform offers a novel approach for rapid AST, demonstrating improved customization and efficiency for AST assays. Its holds potential for addressing multi-drug resistant bacterial strains and accommodating diverse screening scenarios in modern clinical diagnostics.
{"title":"An Integrated Microfluidic Chip for Rapid and Multiple Antimicrobial Susceptibility Testing","authors":"Zirui Pang, Lulu Shi, Mingyu Wang, Jifang Tao","doi":"10.1039/d4an01430h","DOIUrl":"https://doi.org/10.1039/d4an01430h","url":null,"abstract":"The overuse and misuse of antibiotics have caused the development of antimicrobial resistance (AMR), which poses a significant threat to human health. Antimicrobial susceptibility testing (AST) serves as an effective tool for assessing the susceptibility of pathogens infecting patients and guiding the precise use of antibiotics. The conventional AST method, however, is limited by prolonged incubation times and high reagent consumption. In this study, we introduce an integrated microfluidic platform, enabling multiple AST and minimum inhibitory concentration (MIC) determinations after 2 hours incubation. Sample loading is achieved using a self-priming and vacuum-driven approach, enhancing operational feasibility and preventing cross-contamination during reagent pre-coating. Moreover, the use of chips with pre-coated antibiotics minimizes the need for reagent handling off-chip, thereby enhancing the flexibility of the microfluidic device and making the platform easy-to-use. The AST on-chip results for Klebsiella pneumoniae (K. pneumoniae) S1 correlate well with broth dilution methods. This integrated microfluidic platform offers a novel approach for rapid AST, demonstrating improved customization and efficiency for AST assays. Its holds potential for addressing multi-drug resistant bacterial strains and accommodating diverse screening scenarios in modern clinical diagnostics.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"6 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143496157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Oluseyi Olawuyi, James Stewart, Md. Minhas Hossain Hossain Sakib, William Bryant, Mary-Kate Wewers, Noam Lewit, Md Ackas Ali, Md Sajjadur Rahman, Mohammad A Halim
The therapeutic deep eutectic solvent is a new class of deep eutectic solvent (DES), which includes at least an active pharmaceutical ingredient (API) as one of its components. Therapeutic DESs are emerging alternatives that improve the bioavailability, solubility, delivery, and pharmacokinetics properties of drugs. DESs comprise two components, generally hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD), with varying ratios. The interaction chemistry between HBA:HBD in DESs is complex. Moreover, stoichiometry and cluster formation of DES at the molecular level received little attention. Mass spectrometry (MS) is an attractive technique for studying isolated gas phase molecules; however, such investigations have not been implemented for DESs. Compared to other techniques, MS is unique to provide the gas phase stoichiometry, cluster formation, and interaction network between two components of DESs. In addition, computational modeling assists in visualizing the isolated DES clusters and unraveling a deeper understanding of the structure-property relationship. In this study, multi-technique approaches including thermogravimetric (TGA), colorimetric (DSC), spectroscopic (IR and Raman), emerging mass spectrometry, and computational were employed to characterize the menthol:ibuprofen based therapeutics DES. The thermal, colorimetric, and spectroscopies studies showed that hydrogen bonding is the primary factor contributing to DES formation. This study also reported the stable gas phase cluster structure of a menthol: ibuprofen DES using electrospray ionization (ESI) and direct analysis in real time (DART) coupled with mass spectrometry. Subsequently, temperature-dependent DART-MS investigation shows that different temperature conditions impact the formation, intensity of clusters, and the presence of ester impurities. The most intense peak in the ESI-MS and DART-MS spectrum was detected at m/z 363.1, corresponding to the hetero molecular cluster of 1:1 menthol: ibuprofen complex. In addition to the hetero cluster, homo clusters of two-menthol and two-ibuprofen are also detected. Density functional theory (DFT) was employed to investigate the possible gas phase structures of the selected clusters obtained from MS. The DFT results show that hydrogen bonds between the constituents stabilize most of the clusters. MS-guided computational model visualized detailed microstructures and provided insights into the formation mechanism and intermolecular interaction of therapeutic DES.
{"title":"Elucidating Gas Phase Microstructures of Therapeutic Deep Eutectic System","authors":"Oluseyi Olawuyi, James Stewart, Md. Minhas Hossain Hossain Sakib, William Bryant, Mary-Kate Wewers, Noam Lewit, Md Ackas Ali, Md Sajjadur Rahman, Mohammad A Halim","doi":"10.1039/d4an00645c","DOIUrl":"https://doi.org/10.1039/d4an00645c","url":null,"abstract":"The therapeutic deep eutectic solvent is a new class of deep eutectic solvent (DES), which includes at least an active pharmaceutical ingredient (API) as one of its components. Therapeutic DESs are emerging alternatives that improve the bioavailability, solubility, delivery, and pharmacokinetics properties of drugs. DESs comprise two components, generally hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD), with varying ratios. The interaction chemistry between HBA:HBD in DESs is complex. Moreover, stoichiometry and cluster formation of DES at the molecular level received little attention. Mass spectrometry (MS) is an attractive technique for studying isolated gas phase molecules; however, such investigations have not been implemented for DESs. Compared to other techniques, MS is unique to provide the gas phase stoichiometry, cluster formation, and interaction network between two components of DESs. In addition, computational modeling assists in visualizing the isolated DES clusters and unraveling a deeper understanding of the structure-property relationship. In this study, multi-technique approaches including thermogravimetric (TGA), colorimetric (DSC), spectroscopic (IR and Raman), emerging mass spectrometry, and computational were employed to characterize the menthol:ibuprofen based therapeutics DES. The thermal, colorimetric, and spectroscopies studies showed that hydrogen bonding is the primary factor contributing to DES formation. This study also reported the stable gas phase cluster structure of a menthol: ibuprofen DES using electrospray ionization (ESI) and direct analysis in real time (DART) coupled with mass spectrometry. Subsequently, temperature-dependent DART-MS investigation shows that different temperature conditions impact the formation, intensity of clusters, and the presence of ester impurities. The most intense peak in the ESI-MS and DART-MS spectrum was detected at m/z 363.1, corresponding to the hetero molecular cluster of 1:1 menthol: ibuprofen complex. In addition to the hetero cluster, homo clusters of two-menthol and two-ibuprofen are also detected. Density functional theory (DFT) was employed to investigate the possible gas phase structures of the selected clusters obtained from MS. The DFT results show that hydrogen bonds between the constituents stabilize most of the clusters. MS-guided computational model visualized detailed microstructures and provided insights into the formation mechanism and intermolecular interaction of therapeutic DES.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"12 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143486598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kissan D. Achary, Satheesh Natarajan, Aashish Priye
Approximately 9% of the global population lacks access to reliable electricity, and the absence of affordable, electricity-free diagnostic tools hinders early detection of infectious diseases, exacerbating public health burdens in resource-limited settings. We introduce SteamFloc-LAMP – an electricity-free molecular diagnostic platform engineered for instrument-free detection of pathogenic nucleic acid targets. The platform leverages steam-mediated heating from boiling water to sustain the isothermal conditions required for Loop-Mediated Isothermal Amplification (LAMP). Thermal characterization of the steam-mediated heating system identified parameters that enable the consistent maintenance of optimal temperatures for LAMP reactions with minimal fluctuations. Visual end-point detection was achieved through a bridging flocculation mechanism, which exploits the interaction between LAMP amplicons, spermine, and charcoal particles, leading to visible aggregation in positive samples, thus enabling naked-eye detection without the need for specialized equipment or expensive reagents like fluorophores or colorimetric dyes. The SteamFloc-LAMP assay targeted the lipL32 gene, recognized for its exclusivity to pathogenic Leptospira strains. The assay achieved a detection limit of 100 fg of genomic DNA per reaction (∼90 genome copies). Specificity tests using lipL32-specific primers demonstrated the assay's ability to distinguish pathogenic Leptospira accurately, with no cross-reactivity with ligB, ligA, or lipL41 genes found in nonpathogenic strains. A blind test involving DNA extracted from Leptospira reference cultures further validated the assay's diagnostic accuracy, aligning with PCR results. These findings demonstrate the SteamFloc-LAMP assay as a reliable, simple, and cost-effective field deployable diagnostic tool with significant implications for point-of-care detection.
{"title":"A steam-mediated isothermal amplification and flocculation-based detection platform for electricity-free point of care diagnostics","authors":"Kissan D. Achary, Satheesh Natarajan, Aashish Priye","doi":"10.1039/d4an01526f","DOIUrl":"https://doi.org/10.1039/d4an01526f","url":null,"abstract":"Approximately 9% of the global population lacks access to reliable electricity, and the absence of affordable, electricity-free diagnostic tools hinders early detection of infectious diseases, exacerbating public health burdens in resource-limited settings. We introduce SteamFloc-LAMP – an electricity-free molecular diagnostic platform engineered for instrument-free detection of pathogenic nucleic acid targets. The platform leverages steam-mediated heating from boiling water to sustain the isothermal conditions required for Loop-Mediated Isothermal Amplification (LAMP). Thermal characterization of the steam-mediated heating system identified parameters that enable the consistent maintenance of optimal temperatures for LAMP reactions with minimal fluctuations. Visual end-point detection was achieved through a bridging flocculation mechanism, which exploits the interaction between LAMP amplicons, spermine, and charcoal particles, leading to visible aggregation in positive samples, thus enabling naked-eye detection without the need for specialized equipment or expensive reagents like fluorophores or colorimetric dyes. The SteamFloc-LAMP assay targeted the <em>lip</em>L32 gene, recognized for its exclusivity to pathogenic <em>Leptospira</em> strains. The assay achieved a detection limit of 100 fg of genomic DNA per reaction (∼90 genome copies). Specificity tests using <em>lip</em>L32-specific primers demonstrated the assay's ability to distinguish pathogenic <em>Leptospira</em> accurately, with no cross-reactivity with <em>lig</em>B, <em>lig</em>A, or <em>lip</em>L41 genes found in nonpathogenic strains. A blind test involving DNA extracted from <em>Leptospira</em> reference cultures further validated the assay's diagnostic accuracy, aligning with PCR results. These findings demonstrate the SteamFloc-LAMP assay as a reliable, simple, and cost-effective field deployable diagnostic tool with significant implications for point-of-care detection.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"26 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143477477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We reported a novel DNA lesion-modified nanotweezer that underwent conformational switch upon ALKBH enzymes-mediated lesion repair. We demonstrated that the DNA lesion-modified nanotweezer enabled specifically and rapidly monitoring ALKBH2 and ALKBH3 activities and screening inhibitors. Important, this strategy allowed evaluating ALKBH2-associated drug resistance in cells.
{"title":"“Repair and Fold” DNA Nanotweezers for Measuring DNA Alkylation Repair Mediated by ALKBH","authors":"Fengze Jiang, Cui Zhang, Xiangnan Wang, Yuan Yin, Jieling Hong, Hao Tang, Lijuan Tang, Zhenkun Wu","doi":"10.1039/d5an00099h","DOIUrl":"https://doi.org/10.1039/d5an00099h","url":null,"abstract":"We reported a novel DNA lesion-modified nanotweezer that underwent conformational switch upon ALKBH enzymes-mediated lesion repair. We demonstrated that the DNA lesion-modified nanotweezer enabled specifically and rapidly monitoring ALKBH2 and ALKBH3 activities and screening inhibitors. Important, this strategy allowed evaluating ALKBH2-associated drug resistance in cells.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"74 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143477480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Laccase is a kind of copper-containing polyphenol oxidase with similar properties of different sources, of which catalytic reaction requires only oxygen and releases only water. The most widely used method for laccase detection is spectrophotometric assay with limited sensitivity. Herein, we report a highly sensitive fluorescent method for detecting the activity of laccase. This probe shows high selectivity, reliable accuracy as well as a good linear relationship toward different concentrations of laccase. Upon reaction, a new absorption peak appears around 420 nm accompanied by a significant fluorescence increase at 470 nm, which was consistent with the conjugation extension after the polyphenol structure was oxidized to quinone. The effects of different inhibitors and metal ions on the activity of laccase were assessed and this probe was compared with a commercial laccase detection kit showing higher sensitivity. Further, this method was successfully used for the determination of laccase in grape musts with different degrees of fermentation as well as in three kinds of mushroom extracts.
{"title":"A Highly Sensitive Caffeic Acid Fluorescent Probe for Detecting Laccase in Grape Juice and Mushrooms","authors":"Haolin Zhang, Zixu He, Xiaofan Zhang, Xiaohua Li","doi":"10.1039/d5an00071h","DOIUrl":"https://doi.org/10.1039/d5an00071h","url":null,"abstract":"Laccase is a kind of copper-containing polyphenol oxidase with similar properties of different sources, of which catalytic reaction requires only oxygen and releases only water. The most widely used method for laccase detection is spectrophotometric assay with limited sensitivity. Herein, we report a highly sensitive fluorescent method for detecting the activity of laccase. This probe shows high selectivity, reliable accuracy as well as a good linear relationship toward different concentrations of laccase. Upon reaction, a new absorption peak appears around 420 nm accompanied by a significant fluorescence increase at 470 nm, which was consistent with the conjugation extension after the polyphenol structure was oxidized to quinone. The effects of different inhibitors and metal ions on the activity of laccase were assessed and this probe was compared with a commercial laccase detection kit showing higher sensitivity. Further, this method was successfully used for the determination of laccase in grape musts with different degrees of fermentation as well as in three kinds of mushroom extracts.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"5 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143477481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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