This study explores the influence of solar irradiance (Ir) and ambient temperature (T) on photovoltaic (PV) production (P) by combining statistical analysis and deep learning techniques. A strong positive correlation was found between irradiance and PV output with Pearson coefficient (R=0.9211), while temperature exhibited a moderate effect (R=0.4477). A two-way analyze of variance (ANOVA) confirmed the statistical significance of these environmental factors. Furthermore, an autoencoder-based model was developed to capture complex nonlinear relationships and outperformed classical regression models in terms of accuracy and generalization. These results highlight the potential of machine learning methods for improving the understanding and optimization of PV systems in variable climatic conditions. Combines statistical analysis and deep learning to assess PV production factors Employs an autoencoder model to capture nonlinear relationships more effectively The model integrates a new interaction term that increases sensitivity to joint environmental variations.
{"title":"Analysis of the impact of irradiance and temperature on photovoltaic production: A statistical and machine learning approach.","authors":"El Hadji Mbaye Ndiaye, Alphousseyni Ndiaye, Mactar Faye, Daouda Gueye, Amadou Ba, Mamadou Traore","doi":"10.1016/j.mex.2025.103716","DOIUrl":"10.1016/j.mex.2025.103716","url":null,"abstract":"<p><p>This study explores the influence of solar irradiance (Ir) and ambient temperature (T) on photovoltaic (PV) production (P) by combining statistical analysis and deep learning techniques. A strong positive correlation was found between irradiance and PV output with Pearson coefficient (R=0.9211), while temperature exhibited a moderate effect (R=0.4477). A two-way analyze of variance (ANOVA) confirmed the statistical significance of these environmental factors. Furthermore, an autoencoder-based model was developed to capture complex nonlinear relationships and outperformed classical regression models in terms of accuracy and generalization. These results highlight the potential of machine learning methods for improving the understanding and optimization of PV systems in variable climatic conditions. Combines statistical analysis and deep learning to assess PV production factors Employs an autoencoder model to capture nonlinear relationships more effectively The model integrates a new interaction term <math> <mrow><msub><mi>I</mi> <mi>r</mi></msub> <mo>.</mo> <mi>T</mi></mrow> </math> that increases sensitivity to joint environmental variations.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103716"},"PeriodicalIF":1.9,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12664067/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145648957","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}
Pub Date : 2025-11-07DOI: 10.1016/j.mex.2025.103715
Arato Takedachi, Mika Hayashida, Isao Kuraoka
Mismatch repair (MMR) contributes to accurate DNA replication by eliminating mismatched bases during DNA synthesis. Its importance is underscored by Lynch syndrome, a common hereditary colorectal cancer syndrome caused by MMR gene mutations. Therefore, quantification of MMR activity in human cells is important for diagnostic and therapeutic purposes.
To monitor MMR, we established a novel DNA plasmid, pBluescript II NLS-MC-EGFP-tdTomato (pBET2). After processing using our protocol called the oligo swapping method, the mismatch included in the pBET2 enables us to evaluate the MMR proficiency of living cells by detecting specific fluorescent markers. Since the method simply “swaps” short single-stranded DNA to create a single mismatch on a specific site of the plasmid, it is significantly easier and more user-friendly. Briefly, the nicking endonuclease Nt.BbvCI produces a short single-stranded DNA region in the pBET2. Swapping is achieved by filling the region with a mismatched oligonucleotide, and T4 DNA ligase seals the gap. To isolate the mismatch substrate, the restriction enzyme SpeI-HF and T5 exonuclease clean up all contaminants, such as non-mismatch and/or non-covalently closed circular DNA substrates. Finally, the nicking endonuclease Nt.BspQI induces a nick on the final mismatch substrates that enhances the MMR reaction specifically on the target strand.
● This experimental method allows for the easy preparation of mismatched DNA substrates, in contrast to conventionally complex methods.
● This method enables the specific and efficient evaluation of MMR activity in living cells by using the mismatch substrate, a fluorescent marker.
● This method is applicable not only to MMR but also to the preparation of substrates for other DNA repair pathways, making it a valuable resource for both in vitro and in vivo experiments.
错配修复(MMR)通过消除DNA合成过程中不匹配的碱基,有助于准确的DNA复制。Lynch综合征,一种由MMR基因突变引起的常见遗传性结直肠癌综合征,强调了其重要性。因此,人类细胞中MMR活性的定量对诊断和治疗具有重要意义。为了监测MMR,我们建立了一个新的DNA质粒pBluescript II NLS-MC-EGFP-tdTomato (pBET2)。在使用我们称为寡核苷酸交换方法的协议处理后,pBET2中包含的不匹配使我们能够通过检测特定的荧光标记来评估活细胞的MMR熟练程度。由于该方法只是简单地“交换”短单链DNA,在质粒的特定位点上产生单个不匹配,因此它明显更容易和更用户友好。简单地说,核酸内切酶Nt.BbvCI在pBET2中产生短的单链DNA区域。交换是通过用不匹配的寡核苷酸填充该区域来实现的,T4 DNA连接酶密封了间隙。为了分离错配底物,限制酶SpeI-HF和T5外切酶清除所有污染物,如非错配和/或非共价闭合的环状DNA底物。最后,缺口内切酶Nt.BspQI在最终的错配底物上诱导缺口,从而特异性地增强靶链上的MMR反应。●与传统复杂的方法相比,该实验方法可以轻松制备不匹配的DNA底物。●该方法通过使用失配底物(一种荧光标记物),能够对活细胞中的MMR活性进行特异性和高效的评估。●该方法不仅适用于MMR,也适用于其他DNA修复途径的底物制备,是体外和体内实验的宝贵资源。
{"title":"An oligo-swapping method: preparation of mismatch repair-monitoring substrate using a nicking endonuclease","authors":"Arato Takedachi, Mika Hayashida, Isao Kuraoka","doi":"10.1016/j.mex.2025.103715","DOIUrl":"10.1016/j.mex.2025.103715","url":null,"abstract":"<div><div>Mismatch repair (MMR) contributes to accurate DNA replication by eliminating mismatched bases during DNA synthesis. Its importance is underscored by Lynch syndrome, a common hereditary colorectal cancer syndrome caused by MMR gene mutations. Therefore, quantification of MMR activity in human cells is important for diagnostic and therapeutic purposes.</div><div>To monitor MMR, we established a novel DNA plasmid, pBluescript II NLS-MC-EGFP-tdTomato (pBET2). After processing using our protocol called the oligo swapping method, the mismatch included in the pBET2 enables us to evaluate the MMR proficiency of living cells by detecting specific fluorescent markers. Since the method simply “swaps” short single-stranded DNA to create a single mismatch on a specific site of the plasmid, it is significantly easier and more user-friendly. Briefly, the nicking endonuclease Nt.<em>Bbv</em>CI produces a short single-stranded DNA region in the pBET2. Swapping is achieved by filling the region with a mismatched oligonucleotide, and T4 DNA ligase seals the gap. To isolate the mismatch substrate, the restriction enzyme <em>Spe</em>I-HF and T5 exonuclease clean up all contaminants, such as non-mismatch and/or non-covalently closed circular DNA substrates. Finally, the nicking endonuclease Nt.<em>Bsp</em>QI induces a nick on the final mismatch substrates that enhances the MMR reaction specifically on the target strand.</div><div>● This experimental method allows for the easy preparation of mismatched DNA substrates, in contrast to conventionally complex methods.</div><div>● This method enables the specific and efficient evaluation of MMR activity in living cells by using the mismatch substrate, a fluorescent marker.</div><div>● This method is applicable not only to MMR but also to the preparation of substrates for other DNA repair pathways, making it a valuable resource for both <em>in vitro</em> and <em>in vivo</em> experiments.</div></div>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"Article 103715"},"PeriodicalIF":1.9,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145516627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07eCollection Date: 2025-12-01DOI: 10.1016/j.mex.2025.103708
Zsófia Ruszkiczay-Rüdiger, Gábor Csillag, Alexander Wieser, Oscar Marchhart, Mihály Braun, László Fodor
In this study a new approach of cosmogenic radionuclide (CRN) isochron burial dating was developed for locations where the maximum clast size of the sediment does not reach the size of using single cobble samples. This modification makes use of the presumption that different grain size fractions have diverse pre-burial histories and thus are deposited with diverse amount of CRNs. Accordingly, a bulk sample is taken at a certain sediment depth, from which several grain size fractions are selected. These are treated as individual samples for isochron burial age determination. To test our hypothesis, a fluvial sediment succession of the Paleo-Danube River in the Western Pannonian Basin, with an assumed Pliocene age was sampled at two sample depths. The different grain size fractions (0.25-0.5; 1-2; 4-8; 10-20; and 40-60 mm; n=7) provided variable 26Al and 10Be concentrations, supporting the validity of our assumption. The difference between the lowest and largest concentrations was limited making age determination and outlier identification more challenging compared to an isochron with more variable CRN concentrations. After a careful outlier identification, χ2 minimisation inverse modelling yielded 3.9 ± 0.7 Ma as the age of fluvial sedimentation. The offered sampling strategy opens a window towards further application of the CRN isochron burial dating. The burial age of fluvial sediments that were hitherto undatable due to their limited thickness and small clast size can be determined. After the presented adjustment of the sampling strategy the age of deposition can be calculated by the standard ways of isochron burial dating.
{"title":"Testing grain size dependent variability of cosmogenic nuclide concentrations for isochron burial dating of fluvial sediment (Pannonian Basin, Hungary).","authors":"Zsófia Ruszkiczay-Rüdiger, Gábor Csillag, Alexander Wieser, Oscar Marchhart, Mihály Braun, László Fodor","doi":"10.1016/j.mex.2025.103708","DOIUrl":"10.1016/j.mex.2025.103708","url":null,"abstract":"<p><p>In this study a new approach of cosmogenic radionuclide (CRN) isochron burial dating was developed for locations where the maximum clast size of the sediment does not reach the size of using single cobble samples. This modification makes use of the presumption that different grain size fractions have diverse pre-burial histories and thus are deposited with diverse amount of CRNs. Accordingly, a bulk sample is taken at a certain sediment depth, from which several grain size fractions are selected. These are treated as individual samples for isochron burial age determination. To test our hypothesis, a fluvial sediment succession of the Paleo-Danube River in the Western Pannonian Basin, with an assumed Pliocene age was sampled at two sample depths. The different grain size fractions (0.25-0.5; 1-2; 4-8; 10-20; and 40-60 mm; n=7) provided variable <sup>26</sup>Al and <sup>10</sup>Be concentrations, supporting the validity of our assumption. The difference between the lowest and largest concentrations was limited making age determination and outlier identification more challenging compared to an isochron with more variable CRN concentrations. After a careful outlier identification, χ<sup>2</sup> minimisation inverse modelling yielded 3.9 ± 0.7 Ma as the age of fluvial sedimentation. The offered sampling strategy opens a window towards further application of the CRN isochron burial dating. The burial age of fluvial sediments that were hitherto undatable due to their limited thickness and small clast size can be determined. After the presented adjustment of the sampling strategy the age of deposition can be calculated by the standard ways of isochron burial dating.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103708"},"PeriodicalIF":1.9,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12663639/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145648971","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}
Pub Date : 2025-11-07eCollection Date: 2025-12-01DOI: 10.1016/j.mex.2025.103712
Frederik Schaltz-Buchholzer, Ole Bæk, Amrit Singh, Elsi Cá, Isaquel da Silva, Peter Aaby, Christine Stabell Benn, Tobias R Kollmann, Nelly Amenyogbe
The use of Bacillus Calmette-Guérin (BCG) vaccine against tuberculosis (TB) spans more than a century. Besides protection against severe paediatric TB, randomized trials and novel advances within innate immunology documented that BCG has beneficial non-specific effects, providing protection against non-TB infections. Since paediatric intradermal BCG vaccination has proved unable to contain adult pulmonary tuberculosis, several novel TB vaccines are under development, most of which build upon BCG. BCG's status as an essential remedy against TB will therefore be maintained, but despite many decades of near-universal intradermal use, the local responses to BCG in the skin have not been thoroughly elucidated. We therefore developed appropriate methods to capture the localised skin events at the cellular and molecular level after intradermal BCG vaccination. This work informs future studies to identify the immunological events induced following administration of BCG to accelerate development of improved or new vaccines against Mycobacterium tuberculosis (Mtb). We employed advanced technologies such as spatial transcriptomics on skin tissue punch biopsies and cell-free plasma transcriptomics (liquid biopsies) to characterize both the local (in situ) skin and systemic (peripheral blood) response to BCG via:•Whole blood transcriptomics and epigenetic analysis•Blood immune cell characterization•Plasma proteomic and metabolomic analysis.
{"title":"The BCG <i>in situ</i> study - novel techniques applied to a 100-year-old vaccine.","authors":"Frederik Schaltz-Buchholzer, Ole Bæk, Amrit Singh, Elsi Cá, Isaquel da Silva, Peter Aaby, Christine Stabell Benn, Tobias R Kollmann, Nelly Amenyogbe","doi":"10.1016/j.mex.2025.103712","DOIUrl":"10.1016/j.mex.2025.103712","url":null,"abstract":"<p><p>The use of Bacillus Calmette-Guérin (BCG) vaccine against tuberculosis (TB) spans more than a century. Besides protection against severe paediatric TB, randomized trials and novel advances within innate immunology documented that BCG has beneficial non-specific effects, providing protection against non-TB infections. Since paediatric intradermal BCG vaccination has proved unable to contain adult pulmonary tuberculosis, several novel TB vaccines are under development, most of which build upon BCG. BCG's status as an essential remedy against TB will therefore be maintained, but despite many decades of near-universal intradermal use, the local responses to BCG in the skin have not been thoroughly elucidated. We therefore developed appropriate methods to capture the localised skin events at the cellular and molecular level after intradermal BCG vaccination. This work informs future studies to identify the immunological events induced following administration of BCG to accelerate development of improved or new vaccines against <i>Mycobacterium tuberculosis (Mtb)</i>. We employed advanced technologies such as spatial transcriptomics on skin tissue punch biopsies and cell-free plasma transcriptomics (liquid biopsies) to characterize both the local (<i>in situ</i>) skin and systemic (peripheral blood) response to BCG via:•Whole blood transcriptomics and epigenetic analysis•Blood immune cell characterization•Plasma proteomic and metabolomic analysis.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103712"},"PeriodicalIF":1.9,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12681442/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145708018","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}
Pub Date : 2025-11-05eCollection Date: 2025-12-01DOI: 10.1016/j.mex.2025.103711
Elham Faraji, Patricia Cabedo-Sanz, Ajit K Sarmah
Microplastic (MP) contamination in soil media presents growing analytical challenges. We introduce a validated method for MP mass quantification using pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), targeting polyethylene (PE), polypropylene (PP), and polystyrene (PS) in synthetic and environmental soils. The method achieved low limits of detection (0.02-0.44 µg), strong linearity (R² > 0.995), and high recovery rates-86.1% (sandy), 90.7 % (loamy), and 99.6 % (sandy-loam). Cryomilling improved sample homogeneity and quantification accuracy (+3.2 %). Fourier transform infrared spectroscopy (FTIR) was used to confirm polymer identity with >85% match. The protocol was successfully applied to environmental samples from urban and agricultural soils in Auckland, New Zealand, demonstrating its robustness and field applicability. This practical workflow offers a reproducible, high-sensitivity approach suitable for routine microplastics monitoring across diverse soil matrices. Py-GC/MS method achieved over 90 % accuracy for microplastic quantification. Extraction protocols demonstrated recovery efficiencies of up to 99.6 % FTIR complemented Py-GC/MS, confirming polymer identification with >85 % accuracy.
{"title":"A practical method for mass quantification of microplastics in soil media using pyrolysis gas chromatography-mass spectrometry.","authors":"Elham Faraji, Patricia Cabedo-Sanz, Ajit K Sarmah","doi":"10.1016/j.mex.2025.103711","DOIUrl":"10.1016/j.mex.2025.103711","url":null,"abstract":"<p><p>Microplastic (MP) contamination in soil media presents growing analytical challenges. We introduce a validated method for MP mass quantification using pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), targeting polyethylene (PE), polypropylene (PP), and polystyrene (PS) in synthetic and environmental soils. The method achieved low limits of detection (0.02-0.44 µg), strong linearity (R² > 0.995), and high recovery rates-86.1% (sandy), 90.7 % (loamy), and 99.6 % (sandy-loam). Cryomilling improved sample homogeneity and quantification accuracy (+3.2 %). Fourier transform infrared spectroscopy (FTIR) was used to confirm polymer identity with >85% match. The protocol was successfully applied to environmental samples from urban and agricultural soils in Auckland, New Zealand, demonstrating its robustness and field applicability. This practical workflow offers a reproducible, high-sensitivity approach suitable for routine microplastics monitoring across diverse soil matrices. Py-GC/MS method achieved over 90 % accuracy for microplastic quantification. Extraction protocols demonstrated recovery efficiencies of up to 99.6 % FTIR complemented Py-GC/MS, confirming polymer identification with >85 % accuracy.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103711"},"PeriodicalIF":1.9,"publicationDate":"2025-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12664816/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145654482","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}
Pub Date : 2025-11-04eCollection Date: 2025-12-01DOI: 10.1016/j.mex.2025.103709
Seung-Gu Lee
This article provides an update to a previously published data article and associated data set regarding Eu isotope ratio of NIST3117a Eu chemical solution, determined over the period 2017-2024. The Eu isotope ratios were measured via MC-ICP-MS using various Sm isotope pairs -147Sm-149Sm, 147Sm-154Sm, 149Sm-154Sm and 150Sm-154Sm-based on the exponential law over the eight-year period. Measurements were conducted under both wet plasma conditions (using spray chamber) and dry plasma conditions (using Aridus II system). The mean δ¹⁵³Eu values for all Sm isotope pairs are close to 0 ‰, and their 2SD intervals include zero, indicating no systematic bias from the choice of Sm pair or the sample-introduction system. While the precision of δ¹⁵³Eu values for NIST 3117a varies slightly among different Sm isotope pairs, it remains generally consistent. These results demonstrate that Sm-based internal normalization yields robust and reproducible Eu isotope ratios by MC-ICP-MS. The dataset can serve as reference for studying Eu isotope fractionation in geological rocks and meteorites. 36 • No measurable bias in the Eu isotopic composition of the NIST 3117a standard solution is observed with respect to the selection of Sm isotope pairs for normalization or the sample introduction technique.
{"title":"Long term dataset of Eu isotope ratio of NIST3117a Eu standard solution determined via multi-collector inductively coupled plasma mass spectrometry during 2017-2024.","authors":"Seung-Gu Lee","doi":"10.1016/j.mex.2025.103709","DOIUrl":"https://doi.org/10.1016/j.mex.2025.103709","url":null,"abstract":"<p><p>This article provides an update to a previously published data article and associated data set regarding Eu isotope ratio of NIST3117a Eu chemical solution, determined over the period 2017-2024. The Eu isotope ratios were measured via MC-ICP-MS using various Sm isotope pairs -<sup>147</sup>Sm-<sup>149</sup>Sm, <sup>147</sup>Sm-<sup>154</sup>Sm, <sup>149</sup>Sm-<sup>154</sup>Sm and <sup>150</sup>Sm-<sup>154</sup>Sm-based on the exponential law over the eight-year period. Measurements were conducted under both wet plasma conditions (using spray chamber) and dry plasma conditions (using Aridus II system). The mean δ¹⁵³Eu values for all Sm isotope pairs are close to 0 ‰, and their 2SD intervals include zero, indicating no systematic bias from the choice of Sm pair or the sample-introduction system. While the precision of δ¹⁵³Eu values for NIST 3117a varies slightly among different Sm isotope pairs, it remains generally consistent. These results demonstrate that Sm-based internal normalization yields robust and reproducible Eu isotope ratios by MC-ICP-MS. The dataset can serve as reference for studying Eu isotope fractionation in geological rocks and meteorites. 36 • No measurable bias in the Eu isotopic composition of the NIST 3117a standard solution is observed with respect to the selection of Sm isotope pairs for normalization or the sample introduction technique.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103709"},"PeriodicalIF":1.9,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12648585/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145635603","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}
Pub Date : 2025-11-04eCollection Date: 2025-12-01DOI: 10.1016/j.mex.2025.103705
Ntoko M Leakwe, Werner Welthagen, Patricia B C Forbes
Volatile organic compounds (VOCs) are a significant class of air pollutants, as they pose a risk to human health and the environment. Precise quantification of VOCs sampled onto sorbent tubes requires gas phase standards at suitable concentrations. Herein, we present a: - gas standard preparation method for benzene, toluene, ethylbenzene and xylene (BTEX) prepared from liquid standards by means of evaporation and dynamic dilution with nitrogen - demonstration of the applicability of the LiqMix Cascade gas mix system. Repeatability and reproducibility were first successfully demonstrated via direct GC-MS analysis of generated gas mixtures. Then µg levels of generated gas standards were adsorbed onto sorbent tubes employed in active sampling and ng levels were collected onto Radiello cartridges used in passive sampling, followed by liquid extraction or thermal desorption and GC-MS analysis. Accuracy was verified by a BTEX certified reference material (99 % recovery). The LiqMix Cascade gas mix system was shown to produce precise and repeatable concentrations of BTEX gas phase standards for sorbent tube sampling applications for the first time.
{"title":"Production of <i>in situ</i> mixed gas phase volatile organic compound standards for sorbent tube applications.","authors":"Ntoko M Leakwe, Werner Welthagen, Patricia B C Forbes","doi":"10.1016/j.mex.2025.103705","DOIUrl":"https://doi.org/10.1016/j.mex.2025.103705","url":null,"abstract":"<p><p>Volatile organic compounds (VOCs) are a significant class of air pollutants, as they pose a risk to human health and the environment. Precise quantification of VOCs sampled onto sorbent tubes requires gas phase standards at suitable concentrations. Herein, we present a: - gas standard preparation method for benzene, toluene, ethylbenzene and xylene (BTEX) prepared from liquid standards by means of evaporation and dynamic dilution with nitrogen - demonstration of the applicability of the LiqMix Cascade gas mix system. Repeatability and reproducibility were first successfully demonstrated via direct GC-MS analysis of generated gas mixtures. Then µg levels of generated gas standards were adsorbed onto sorbent tubes employed in active sampling and ng levels were collected onto Radiello cartridges used in passive sampling, followed by liquid extraction or thermal desorption and GC-MS analysis. Accuracy was verified by a BTEX certified reference material (99 % recovery). The LiqMix Cascade gas mix system was shown to produce precise and repeatable concentrations of BTEX gas phase standards for sorbent tube sampling applications for the first time.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103705"},"PeriodicalIF":1.9,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12651840/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145635601","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}
This study proposes a real-time tool wear monitoring approach for dry milling of AISI 304 stainless steel using a Microflown PU sensor and the I-Kaz™ statistical feature. A Taguchi L18 orthogonal array was adopted to optimize cutting speed, feed rate, depth of cut and tool type (uncoated and AlCrN-coated WC-Co inserts) based on surface roughness and flank wear responses. ANOVA revealed that tool type was the most significant factor, contributing approximately 85 % to flank wear variation, followed by depth of cut and cutting speed. The optimal combination: AlCrN-coated insert, cutting speed of 1250 rpm, depth of cut of 0.50 mm and feed rate of 0.04 mm/rev minimized both responses. The proposed I-Kaz-based monitoring approach established a strong inverse power-law relationship between the I-Kaz coefficient and flank wear Z2α =a(VB)-n, achieving R2>0.96, indicating high accuracy and stability across repetitions.•This study introduces the first application of a Microflown PU sensor for near-field acoustic monitoring in milling operations.•The I-Kaz™ feature is demonstrated as a computationally efficient and accurate method for real-time tool wear prediction.•The research integrates cutting parameter optimization and predictive monitoring within a single experimental framework.
{"title":"I-Kaz methodology for predicting tool life of AlCrN-Coated WC-Co inserts in the machining of AISI 304 steel.","authors":"Mogana Priya Chinnasamy, Rajasekar Rathanasamy, Swetha R Kumar, Sathish Kumar Palaniappan","doi":"10.1016/j.mex.2025.103706","DOIUrl":"https://doi.org/10.1016/j.mex.2025.103706","url":null,"abstract":"<p><p>This study proposes a real-time tool wear monitoring approach for dry milling of AISI 304 stainless steel using a Microflown PU sensor and the I-Kaz™ statistical feature. A Taguchi L18 orthogonal array was adopted to optimize cutting speed, feed rate, depth of cut and tool type (uncoated and AlCrN-coated WC-Co inserts) based on surface roughness and flank wear responses. ANOVA revealed that tool type was the most significant factor, contributing approximately 85 % to flank wear variation, followed by depth of cut and cutting speed. The optimal combination: AlCrN-coated insert, cutting speed of 1250 rpm, depth of cut of 0.50 mm and feed rate of 0.04 mm/rev minimized both responses. The proposed I-Kaz-based monitoring approach established a strong inverse power-law relationship between the I-Kaz coefficient and flank wear Z<sub>2</sub> <sup>α</sup> =<i>a</i>(VB)<sup>-n</sup>, achieving R<sup>2</sup>>0.96, indicating high accuracy and stability across repetitions.•This study introduces the first application of a Microflown PU sensor for near-field acoustic monitoring in milling operations.•The I-Kaz™ feature is demonstrated as a computationally efficient and accurate method for real-time tool wear prediction.•The research integrates cutting parameter optimization and predictive monitoring within a single experimental framework.</p>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"103706"},"PeriodicalIF":1.9,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12651424/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145635685","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}
The aim of the present study was to identify the native Spirulina species from Caspian Sea and optimize the growth conditions of this microalgae in order to reduce costs and increase the benefits of its production. In this study, water sampling was carried out from a depth of 5 meters on the coast of south Caspian Sea (Iran) in 2023–2024. After observing the Spirulina genus, its microalgae were isolated and purified using the dilution method, and finally pure single colonies were obtained. Initial identification was carried out using a light microscope and valid identification keys. Molecular identification and 16 SrRNA sequencing were used to confirm and verify it. The results showed that the studied species has the least genetic distance from the microalgae Spirulina laxissima. Then, the effect of three physical factors, temperature (27–29 °C), light (3000–3500 lx) and pH (9–11), on its biomass production rate was investigated using the response surface methodology. The results obtained from the design of this experiment showed that the biomass amount on the seventh day was 0.382 (g/L) with pH=10, light 3250 lx and temperature 29 °C and on the fourteenth day the biomass amount reached 0.839 (g/L) and pH=10, light 3250 lx, temperature 29 °C.
•
Isolation of microalgae Spirulina laxissima from Caspian Sea.
•
Molecular identification Spirulina laxissima.
•
Optimizing of Spirulina laxissima culture with physical changes (temperature, light, pH).
{"title":"Isolation, identification and optimization of biomass production of native Spirulina species from South of the Caspian Sea","authors":"Matin Shakoori , Reza Safari , Asieh Makhlough , Tahereh Bagheri","doi":"10.1016/j.mex.2025.103707","DOIUrl":"10.1016/j.mex.2025.103707","url":null,"abstract":"<div><div>The aim of the present study was to identify the native Spirulina species from Caspian Sea and optimize the growth conditions of this microalgae in order to reduce costs and increase the benefits of its production. In this study, water sampling was carried out from a depth of 5 meters on the coast of south Caspian Sea (Iran) in 2023–2024. After observing the Spirulina genus, its microalgae were isolated and purified using the dilution method, and finally pure single colonies were obtained. Initial identification was carried out using a light microscope and valid identification keys. Molecular identification and 16 SrRNA sequencing were used to confirm and verify it. The results showed that the studied species has the least genetic distance from the microalgae <em>Spirulina laxissima</em>. Then, the effect of three physical factors, temperature (27–29 °C), light (3000–3500 lx) and pH (9–11), on its biomass production rate was investigated using the response surface methodology. The results obtained from the design of this experiment showed that the biomass amount on the seventh day was 0.382 (g/L) with pH=10, light 3250 lx and temperature 29 °C and on the fourteenth day the biomass amount reached 0.839 (g/L) and pH=10, light 3250 lx, temperature 29 °C.<ul><li><span>•</span><span><div>Isolation of microalgae <em>Spirulina laxissima</em> from Caspian Sea.</div></span></li><li><span>•</span><span><div>Molecular identification <em>Spirulina laxissima</em>.</div></span></li><li><span>•</span><span><div>Optimizing of <em>Spirulina laxissima</em> culture with physical changes (temperature, light, pH).</div></span></li></ul></div></div>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"Article 103707"},"PeriodicalIF":1.9,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145516624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-01DOI: 10.1016/j.mex.2025.103703
Hui Zhi Low, Christina Böhnlein, Charles M.A.P. Franz
One-step, reverse transcriptase-quantitative PCR (RT-qPCR) is the primary method for detecting foodborne viruses in food matrices. The ISO 15216-2:2019 serves as the international standard for detecting human norovirus GI, GII, and hepatitis A virus. Some food matrices, such as berries, tend to co-purify PCR inhibitors with viral RNA, which can lead to false-negative results. To prevent this, the protocol includes extensive control approaches. However, the high cost of commercial RT-qPCR kits makes large-scale virus testing expensive and inaccessible. To address this, we developed an in-house, one-step RT-qPCR mix using commercial, next-generation enzymes with improved resistance to PCR inhibitors and with enhanced performance. The in-house mix offers a more cost-effective alternative to expensive and outdated commercial mixes. In this paper, we describe:
• the development of an in-house, one-step multiplexable RT-qPCR protocol and optimization process as a reference for laboratories seeking to develop their own in-house protocols.
• altered and optimized, previously described primers for the MS2 virus, further improving the efficiency of its detection and its reliability as a process control virus.
{"title":"Development of an in-house, one-step RT-qPCR mix and optimized MS2 detection primers for hepatitis A virus and norovirus detection in berries","authors":"Hui Zhi Low, Christina Böhnlein, Charles M.A.P. Franz","doi":"10.1016/j.mex.2025.103703","DOIUrl":"10.1016/j.mex.2025.103703","url":null,"abstract":"<div><div>One-step, reverse transcriptase-quantitative PCR (RT-qPCR) is the primary method for detecting foodborne viruses in food matrices. The ISO 15216-2:2019 serves as the international standard for detecting human norovirus GI, GII, and hepatitis A virus. Some food matrices, such as berries, tend to co-purify PCR inhibitors with viral RNA, which can lead to false-negative results. To prevent this, the protocol includes extensive control approaches. However, the high cost of commercial RT-qPCR kits makes large-scale virus testing expensive and inaccessible. To address this, we developed an in-house, one-step RT-qPCR mix using commercial, next-generation enzymes with improved resistance to PCR inhibitors and with enhanced performance. The in-house mix offers a more cost-effective alternative to expensive and outdated commercial mixes. In this paper, we describe:</div><div>• the development of an in-house, one-step multiplexable RT-qPCR protocol and optimization process as a reference for laboratories seeking to develop their own in-house protocols.</div><div>• altered and optimized, previously described primers for the MS2 virus, further improving the efficiency of its detection and its reliability as a process control virus.</div></div>","PeriodicalId":18446,"journal":{"name":"MethodsX","volume":"15 ","pages":"Article 103703"},"PeriodicalIF":1.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145516626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}