{"title":"Predictive performance of a centrosome-associated prognostic model in prognosis and immunotherapy of lung adenocarcinoma.","authors":"Feng Yan, Qian Guo, Rongbing Zheng, Jiongming Ying","doi":"10.1016/j.ab.2024.115731","DOIUrl":"10.1016/j.ab.2024.115731","url":null,"abstract":"<p><p>In recent years, mounting investigations have highlighted the pivotal role of centrosomes in cancer progression. In this study, we employed bioinformatics and statistics to establish a 13-centrosome-associated gene prognostic model for lung adenocarcinoma (LUAD) utilizing transcriptomic data from TCGA. Based on the Riskscore, patients were stratified into high- and low-risk groups. Through survival analysis and receiver operating characteristic curve analysis, our model demonstrated a consistent and robust prognostic capacity, which was further validated using the GEO database. Univariate/multivariate Cox regression analyses identified our model as an independent prognostic factor for LUAD patients. Subsequently, immunoinfiltration analysis showed that immune cell infiltration levels of aDCs, iDCs, Mast cells, and Neutrophils, as well as immune functionalities such as HLA, Type I IFN Response and Type II IFN Response, were markedly reduced in the high-risk group compared to the low-risk group. Finally, we conducted a drug screening to identify potential treatments for patients with different prognoses. We utilized the GDSC database and molecular docking techniques to identify small molecule compounds targeting the prognostic genes. In conclusion, our prognostic model exhibits robust and reliable predictive capability, and it may have important clinical implications in guiding treatment decisions for LUAD patients.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115731"},"PeriodicalIF":2.6,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chengxin Su , Xiaoling Liu , Ke Zhang , Bing Jiang , Jiashuai Hu , Mei Li , Lin Cheng , Hongbing Luo , Wanchen Xie , Cheng Liu , Liangqian Fan , Wei Chen , Xiaohong Zhang
{"title":"A molecularly imprinted electrochemical sensor based on in-situ polymerization for rapid and selective detection of tonalide in aqueous environment","authors":"Chengxin Su , Xiaoling Liu , Ke Zhang , Bing Jiang , Jiashuai Hu , Mei Li , Lin Cheng , Hongbing Luo , Wanchen Xie , Cheng Liu , Liangqian Fan , Wei Chen , Xiaohong Zhang","doi":"10.1016/j.ab.2024.115730","DOIUrl":"10.1016/j.ab.2024.115730","url":null,"abstract":"<div><div>Given the adverse effects of tonalide (AHTN) on aquatic organisms and humans, coupled with the limitations of current detection methods, which are time-consuming, require expensive equipment and complicated sample preparation procedures, there is a clear need to develop a new technique for detecting AHTN that is highly sensitive, rapid, cost-effective and efficient. In this study, a new simple electrochemical sensor for the determination of AHTN in aqueous environments was developed for the first time through the in-situ polymerization of an AHTN-imprinted polymer on the surface of a graphene (G)-modified carbon electrode (GCE). Following a series of comparative tests, including cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM), the novel AHTN molecularly imprinted sensor (AHTN-MIP/G/GCE) has been demonstrated to be an effective tool for monitoring AHTN. The results demonstrate that the linear detection range of the current response of the AHTN-MIP/G/GCE <sup>1</sup>electrode to AHTN was 0.01 μM–4 μM (i.e., 2.584 μg/L-1033.6 μg/L), with a detection limit of 2.3 × 10⁻⁹ M (i.e., 594.32 ng/L), following the optimization of the experimental conditions. Furthermore, the new sensor was successfully employed for the detection of AHTN in water samples, with recoveries of 97.1%–108.2 % with the added standards. Consequently, the new electrochemical sensor demonstrated good stability and acceptable reproducibility. This study provides a new method for the future detection of AHTN in the aqueous environment.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115730"},"PeriodicalIF":2.6,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142758892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Survey of chemical unfolding complexity as a unique stability assessment assay for monoclonal antibodies.","authors":"J Alaina Floyd, Jeremy M Shaver","doi":"10.1016/j.ab.2024.115729","DOIUrl":"10.1016/j.ab.2024.115729","url":null,"abstract":"<p><p>Seventy-two intentionally sequence-diverse antibody variable regions were selected, expressed as IgG1 antibodies, and evaluated by chemical unfolding to survey the complexities of denaturant induced unfolding behavior. A two-transition fit well described the curves and uncovered a wide range of sensitivities to denaturant. Four general types of unfolding curves were observed: balanced traces (each transition responsible for half of the total unfolding curve), low-unfolding traces (first transition is a majority of the unfolding curve), high-unfolding traces (the second transition is the majority of the unfolding curve), and coincident traces (the two transitions are found close to each other, approximating a single transition). The complexity of the data from this survey indicates that focusing on the first inflection point or fitting a single transition model is likely an over-simplistic method for measuring stability by the chemical unfolding assay. Additionally, other conformational assays, such as thermal and low pH unfolding, showed no correlation with the chemical unfolding results, indicating that each of these assays provide alternate information on the different pathways of antibody conformational stability. These results provide a basis for beginning to better connect unfolding behavior to other physical phenotypic behaviors and production process behaviors.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115729"},"PeriodicalIF":2.6,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Electrochemical sensing of dopamine using nanostructured silver chromate: Development of an IoT-integrated sensor.","authors":"Lungelo Mgenge, Chandan Saha, Pooja Kumari, Sarit K Ghosh, Harishchandra Singh, Kaushik Mallick","doi":"10.1016/j.ab.2024.115726","DOIUrl":"10.1016/j.ab.2024.115726","url":null,"abstract":"<p><p>Dopamine, one of the most important neurotransmitters, plays a crucial role in the functions of human metabolism, as well as the cardiovascular, central nervous and hormonal systems. This study focuses on the synthesis of nanostructured silver chromate and their application in dopamine sensing. The nanoparticles were synthesized using a complexation-mediated route using aminosalicylic acid as a stabilizer, resulting in uniform particles ranging from 3 to 15 nm in size. The synthesized silver chromate was characterized using X-ray diffraction, transmission electron microscopy and X-ray photoelectron spectroscopy techniques. Electrochemical studies revealed that the silver chromate exhibit excellent catalytic activity for the detection of dopamine. The electroanalysis provided the selective recognition of dopamine with the limit of detection of 1.05 μM and sensitivity of 2.68 μA μM<sup>-1</sup> cm<sup>-2</sup> in a linear range of 5-45 μM. Additionally, a portable, IoT (internet of things)-integrated sensor based on the synthesized silver chromate was developed using Arduino Uno R4 Wi-Fi module, enabling real-time monitoring of dopamine with data transmission to a cloud platform.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115726"},"PeriodicalIF":2.6,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Keyang Lai , Jun Xu , Kai Luo , Min Xie , Yuan Chen , Fan Li , Yaomin Zhou , Lihui Gong , Yonghua Xiong , Weihua Lai
{"title":"Lateral flow immunoassay based on aggregation induced emission nanobeads for the sensitive and accurate detection of chloramphenicol in pig hair","authors":"Keyang Lai , Jun Xu , Kai Luo , Min Xie , Yuan Chen , Fan Li , Yaomin Zhou , Lihui Gong , Yonghua Xiong , Weihua Lai","doi":"10.1016/j.ab.2024.115728","DOIUrl":"10.1016/j.ab.2024.115728","url":null,"abstract":"<div><div>Chloramphenicol (CAP) is a once widely used antibiotic, which is able to cause great harm to human health, is banned in some countries or organizations such as China, USA and the European Union for animal breeding. Because CAP in pig hair degraded slower and had amount of residues, pig hair could be used as the target to detect CAP residues. In this study, a competitive lateral flow immunoassay (LFIA), whose label was aggregation induced emission fluorescent nanobeads (AIEFN), was firstly developed for the detection of CAP in pig hair samples. It exhibited a low limit of detection (0.001 μg/kg) and a broad linear range (0.0025–0.32 μg/kg). The recovery rate and coefficient variation with spiked pig hair samples were 88.50%–106.17 % and 1.01%–7.37 %, which indicated good accuracy of this assay. The result of AIEFN-LFIA for the detection of CAP in pig hair was consistent with that of liquid chromatography-mass spectrometry. This assay was rapid with the total detection duration of about 20 min. AIEFN-LFIA was able to be used for rapid, sensitive, accurate and convenient detection of CAP residues in pig hair samples.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"698 ","pages":"Article 115728"},"PeriodicalIF":2.6,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A sandwich ELISA for the quantification of the anticancer peptide CIGB-552 in human plasma.","authors":"Nivaldo Angel Gómez Hernández, Gilda Lemos Pérez, Amalia Vazquez Arteaga, Hilda Elisa Garay Pérez, Brizaida Oliva Arguellez, Ania Cabrales Rico, Airela Llamo Guardia, Julio Raúl Fernández Massó","doi":"10.1016/j.ab.2024.115725","DOIUrl":"10.1016/j.ab.2024.115725","url":null,"abstract":"<p><p>CIGB-552 is a synthetic anticancer peptide that has been evaluated in vitro and in vivo in lung and colon cancer models. To optimize therapy in the clinic, pharmacokinetic studies are necessary. Previously, a sandwich-type enzyme-linked immunosorbent assay (ELISA) had been developed by our working group for the quantification of CIGB-552 in biological matrices. The objective of this work was to carry out the full validation of the ELISA to support its application in clinical trials. First, we obtained a polyclonal antibody specific for CIGB-552 and with purity greater than 95 %. The lower limit of quantification and the upper limit of quantification were 3125 ng/ml and 200 ng/ml, respectively. The method is exact and precise in the quantification of the peptide with relative error and coefficient of variation values less than 20 %. The ELISA is specific in the presence of CIGB-552 metabolites in the sample, and also presents robustness to certain protocol variations. In summary, the validated ELISA meets the requirements for its application in upcoming clinical trials as part of pharmacokinetic studies.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115725"},"PeriodicalIF":2.6,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Seiji Kubo, Keito Amai, Fumitaka Nakano, Jin Tanaka, Hideki Niimi
{"title":"High-throughput sperm screening using one-step RT-qPCR: Improvement and re-evaluation.","authors":"Seiji Kubo, Keito Amai, Fumitaka Nakano, Jin Tanaka, Hideki Niimi","doi":"10.1016/j.ab.2024.115727","DOIUrl":"10.1016/j.ab.2024.115727","url":null,"abstract":"<p><p>Sperm identification is crucial in sexual assault cases. While microscopic analysis is the gold standard for sperm detection, it is a laborious procedure even for trained personnel. Reverse transcription-quantitative real-time PCR (RT-qPCR) can enhance the screening by detecting sperm-specific mRNA markers, such as protamine 2 (PRM2). This study aimed to develop a one-step RT-qPCR assay targeting PRM2 mRNA. Our assay was capable of detecting as low as 0.01 μL of semen with high specificity and demonstrated successful detection of PRM2 mRNA in simulated-case samples. Owing to the simple workflow involved, our assay requires <30 min for RNA extraction and <60 min for RT-qPCR. Our assay enables high-throughput sperm screening and offers a promising strategy for enhancing the workflow of sexual assault cases.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115727"},"PeriodicalIF":2.6,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142749598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Comparative validation of UV-spectrophotometry and RP-HPLC methods for cefixime and moxifloxacin analysis","authors":"Mahesh Chaudhari , Parul K. Parmar , Kiran Dudhat","doi":"10.1016/j.ab.2024.115724","DOIUrl":"10.1016/j.ab.2024.115724","url":null,"abstract":"<div><h3>Aim</h3><div>This study presents the development and validation of UV-spectrophotometry and RP-HPLC methods for the simultaneous quantification of Cefixime Trihydrate (CEFI) and Moxifloxacin Hydrochloride (MOXI) in pharmaceutical formulations.</div></div><div><h3>Methodology</h3><div>Two UV-spectrophotometric methods, including the absorbance ratio (Q-Absorption) and First Order Derivative Spectroscopy, were developed and validated for their linearity, precision, accuracy, and sensitivity. Additionally, a robust RP-HPLC method using a C18 column and optimized mobile phase was employed for efficient separation and simultaneous estimation of CEFI and MOXI. All methods were validated in accordance with ICH guidelines, with system suitability parameters confirming the reliability of the RP-HPLC method for routine analysis.</div></div><div><h3>Results</h3><div>The absorbance ratio and First Order Derivative methods demonstrated low %R.S.D values, high accuracy, and satisfactory sensitivity for both drugs. Similarly, the RP-HPLC method achieved high resolution, precision, and robustness. Statistical analysis through ANOVA revealed no significant differences between the methods in terms of accuracy and precision. The methods were applied to analyze marketed formulations, further confirming their applicability in routine quality control.</div></div><div><h3>Conclusion</h3><div>In conclusion, the validated methods provide accurate, precise, and sensitive techniques for the simultaneous estimation of CEFI and MOXI, making them suitable for pharmaceutical quality control and regulatory compliance.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115724"},"PeriodicalIF":2.6,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142738136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Validation of assay for measuring acetyl-coenzyme a carboxylase activity in grasses using malachite green","authors":"Yoshinobu Jin","doi":"10.1016/j.ab.2024.115723","DOIUrl":"10.1016/j.ab.2024.115723","url":null,"abstract":"<div><div>Acetyl-CoA carboxylase (ACCase) is one of the most important enzymes as a herbicide target in gramineous plant species, however, assay methods for the enzyme are primarily limited to those using radioisotopes (RI). Typically, the measurement method that uses RI necessitates specialized facilities and equipment, and involves complex procedures throughout the experiment. As another method for detecting ACCase activity, the colorimetric method using malachite green (MG) is also known. However, reports on this method are limited, and information regarding the simplicity of the procedure and the scope of its application remains unclear. To better understand the method using MG and to develop a simpler assay method, crude enzymes extracted from various target-site resistant (TSR) biotypes of blackgrass (<em>Alopecurus myosuroides</em>) were examined in enzyme inhibition tests. As a result, this method was able to accurately detect the relationship between the chemical classes of ACC inhibitors and cross-resistance to specific TSRs. Moreover, the ACCase activity of other grass species was also examined using this method. By using crude enzymes from various species and a commercially available phosphatase kit containing MG, ACCase activity was detectable with good accuracy. In addition, enzyme inhibition studies using ACCase inhibiting herbicides revealed that this method reproduced results similar to those obtained with the RI method. The Z′-factor, an index of high-throughput screening (HTS), was around 0.7, indicating that it is an excellent screening system. These results suggest that the assay method using MG is very simple, labor-saving, and accurate with a throughput much higher than that of the existing RI method. Therefore, it is strongly suggested that the method could replace the RI method in most cases. These results indicate that it is applicable to HTS for ACCase inhibitors.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"697 ","pages":"Article 115723"},"PeriodicalIF":2.6,"publicationDate":"2024-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142715228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"In vitro selection of dye-fluorescence-enhancing peptide aptamer by cDNA display.","authors":"Takashi Kubo, Tomoyuki Koike, Tomoki Ouchi, Nayab Khaliq, Eita Sasaki, Kouichi Kuroda, Mitsuyoshi Ueda, Kenjiro Hanaoka, Naoto Nemoto","doi":"10.1016/j.ab.2024.115722","DOIUrl":"10.1016/j.ab.2024.115722","url":null,"abstract":"<p><p>Although Green Fluorescent Protein (GFP) is useful and most widely used, steric hindrance due to its size and the time required for chromophore formation are complications. However, it is difficult to form chromophores with peptides to reduce the molecular weight. Therefore, we focused on peptides that can become fluorescent by binding to dyes. In this study, a novel dye-fluorescence-enhancing peptide aptamer was selected by the cDNA display method, which was confirmed by the yeast surface display method. This peptide aptamer binds to the non-fluorescent dye QSY®9 and enhances its fluorescence by preventing rotation of its benzene sulfone group. The method described in this paper should enable the development of new cell imaging methods using non-fluorescent dyes and peptides.</p>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":" ","pages":"115722"},"PeriodicalIF":2.6,"publicationDate":"2024-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142709031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}