Article
Chemistry, Analytical
Linlin Zhuang, Jianbo Yang, Chunlei Song, Li Sun, Bin Zhao, Qiuping Shen, Xiyan Ren, Hongjing Shi, Yu Zhang, Mengling Zhu
Summary: An accurate and practical graphene oxide-based method, called GO-ASEA, was developed for rapid and highly sensitive detection of African swine fever virus (ASFV). This novel assay showed excellent specificity and selectivity, with a low detection limit, and demonstrated good clinical applicability. The GO-ASEA method has the potential to serve as a robust tool in epidemic prevention and point-of-care diagnosis.
ANALYTICAL METHODS
(2022)
Article
Chemistry, Analytical
Kangning Wang, Limin He, Andreas Manz, Wenming Wu
Summary: Traditional methods for manufacturing microfluidic chips face challenges in producing complex topologies and three-dimensional microchannels, relying on expensive equipment and complex processes. The proposed low-cost approach of sewing chip pipes to substrates with an embroidery machine offers a more efficient alternative, eliminating the need for specialized equipment and skills.
ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Analytical
Pieter Berden, Rodrigo S. Wiederkehr, Liesbet Lagae, Jan Michiels, Tim Stakenborg, Maarten Fauvart, Willem Van Roy
Summary: We developed an analytical model that explains the relationship between amplification efficiency, template accessibility, and rain in dPCR assays. Through Monte Carlo simulations, we found that reduced amplification efficiency leads to broader threshold cycle distributions, while template inaccessibility results in an exponential tail rain pattern.
ANALYTICAL CHEMISTRY
(2022)
Article
Biotechnology & Applied Microbiology
A. Troscianczyk, A. Nowakiewicz, S. Gnat, D. Lagowski, M. Osinska
Summary: This study aimed to assess resistance and virulence of Enterococcus faecalis isolated from the gastrointestinal tract of dogs and cats, analyze their genotypic variability, and estimate the correlation between antimicrobial resistance, virulence determinants, and genotypic profiles. The results showed a high percentage of resistant strains to various antibiotics, high heterogeneity of genotypic profiles among E. faecalis, and the presence of common virulence genes such as efaAfs, cpd, ccf, and cob. Companion animals were confirmed to be a reservoir of E. faecalis carrying resistance and virulence determinants.
JOURNAL OF APPLIED MICROBIOLOGY
(2021)
Article
Chemistry, Analytical
Xinglu Jiang, Beibei Xie, Kangjing Li, Fengyuan Zhou
Summary: This study achieved controlled release of Taq DNA polymerase in qPCR by introducing BSA hydrogel, resulting in over 20 cycles of linear amplification. The slope of linear amplification was found to have a significant negative correlation with Ct values, and an exponential equation between Ct values and the slopes was fitted by varying primer concentrations. The slopes and Ct values of each qPCR system were corrected using this equation to ensure consistency in amplification efficiency.
ANALYTICAL CHEMISTRY
(2023)
Article
Multidisciplinary Sciences
Muhammad Asif, Hamid Anees Siddiqui, Rubab Zahra Naqvi, Imran Amin, Shaheen Asad, Zahid Mukhtar, Aftab Bashir, Shahid Mansoor
Summary: The article discusses the cultivation of Bt cotton expressing Cry1Ac in Pakistan and the development of insect resistant NIBGE-1601 cotton harboring double gene Cry1Ac-Cry2Ab construct. Event characterization of NIBGE-1601 was conducted using PCR technology. The development of highly specific event detection primers for NIBGE-1601 will aid in breeding new cotton varieties resistant to cotton bollworms.
SCIENTIFIC REPORTS
(2021)
Article
Biochemical Research Methods
Jinrong Shen, Jihong Zheng, Zhenqing Li, Yourong Liu, Fengxiang Jing, Xinjun Wan, Yoshinori Yamaguchi, Songlin Zhuang
Summary: The study introduces an effective technique for the absolute quantification of target nucleic acids, with self-developed software controlling functional units to achieve high-resolution one-time gene chip imaging, speeding up the lab-to-application process.
Article
Infectious Diseases
Axel Ursenbach, Frederic Schramm, Francois Severac, Yves Hansmann, Nicolas Lefebvre, Yvon Ruch, Xavier Argemi
Summary: 16S PCR sequencing has a significant clinical impact on patient management, particularly for suspected CSF infections, patients with culture-negative samples, and those with previous antibiotic treatments.
BMC INFECTIOUS DISEASES
(2021)
Article
Chemistry, Analytical
Bobin Lee, Yeonghun Lee, Seung-Mo Kim, Kihyeun Kim, Min-Gon Kim
Summary: The incidence of deadly infectious diseases has been increasing worldwide in recent years, leading to a high demand for rapid and accurate nucleic acid diagnostic systems. In this study, a rapid membrane-based photothermal polymerase chain reaction (PCR) system was developed, allowing for quantitative fluorescence detection. This system overcomes the limitations of conventional PCR and offers user-friendly micro-scale solution control and sample loading. The reaction pad is coated with an Au nanostructure that acts as a light-to-heat converter for thermocycling. The reaction solution is heated using a 785 nm light source, and the temperature is cycled 25 times between 63°C and 95°C within 6 minutes. The amplified samples on the membrane are then quantitatively analyzed by measuring the fluorescence signal. This membrane-based PCR approach can be easily integrated with various sample pre-treatment processes, making it a promising all-in-one molecular diagnosis platform.
SENSORS AND ACTUATORS B-CHEMICAL
(2022)
Article
Biochemical Research Methods
Weidu Song, Chuanhao Zhang, Huichao Lin, Taiyi Zhang, Haixia Liu, Xiaowen Huang
Summary: The proposed real-time rotary PCR reaction system (RRP) meets the requirements of low cost, low-power consumption, potential for miniaturization, and portability. Experimental and theoretical results demonstrate its feasibility in the rapid quantitative detection of Pseudomonas aeruginosa and have broad application prospects in bacteria detection and food safety.
Article
Biochemical Research Methods
Shaolei Huang, Yiquan An, Bangchao Xi, Xianglian Gong, Zhongfu Chen, Shan Shao, Shengxiang Ge, Jun Zhang, Dongxu Zhang, Ningshao Xia
Summary: Nucleic acid detection is a method that directly identifies the presence of pathogenic microorganisms and has advantages of high sensitivity, commendable specificity, and a short window period. A real-time PCR system based on multiple temperature zones was proposed to solve the time-consuming problem, enabling fast detection speed and high sensitivity under extreme conditions.
Article
Food Science & Technology
Mohd Izhar Ariff Mohd Kashim, Alia Aryssa Abdul Haris, Nur Asmadayana Hasim, Sahilah Abd Mutalib, Nurina Anuar
Summary: The aim of this research was to determine the halal status of cultured meat by detecting species-specific DNA of bovine serum. The results revealed that bovine DNA was detected in all samples tested, indicating that perfect transformation is not possible and is prohibited according to Shariah law.
Article
Infectious Diseases
Vanesa Anton-Vazquez, Rafal Dworakowski, Antonio Cannata, George Amin-Youssef, Margaret Gunning, Alexandros Papachristidis, Phil MacCarthy, Max Baghai, Ranjit Deshpande, Habib Khan, Jonathan Byrne, Amanda Fife
Summary: This study assessed the impact of 16S rDNA PCR in the aetiological diagnosis of culture-negative infective endocarditis (IE) patients. The results showed that 16S rDNA PCR significantly improved the detection and identification of bacterial nucleic acid in heart valve tissue compared to valve culture alone. Therefore, 16S rDNA PCR may be a useful diagnostic tool for culture-negative IE, but further studies are needed to evaluate its clinical impact.
Article
Multidisciplinary Sciences
Dina M. Metwally, Reem Alajmi, Muslimah N. Alsulami, Isra M. Al-Turaiki, Rewaida Abdel-Gaber, Afrah F. Alkhuriji, Haleema H. Albohiri, Khalil Mohamed, Hanadi B. Baghdadi, Manal F. El-Khadragy, Guillermo T. Isaias, Saeed El-Ashram
Summary: The study found that Theileria spp. were detected in 57.8% of sheep and 51.9% of goats in Jeddah, Saudi Arabia. Phylogenetic analysis of the sequence data revealed that the T. ovis identified in this study were closely related to an isolate from Turkey, with pairwise identity ranging from 84.4% to 99.8% and coverage ranging from 52.35% to 99.79%.
Review
Medicine, Legal
Kamayani Vajpayee, Hirak Ranjan Dash, Prakshal B. Parekh, Ritesh K. Shukla
Summary: DNA typing technology is widely used in criminal investigations, often utilizing STR profiles to identify suspects. In some cases, mtDNA and Y STR analyses are also considered. Forensic scientists provide inclusion, exclusion, and inconclusive opinions based on the DNA profiles generated. The presence of PCR inhibitors in the sample leads to inconclusive results, causing difficulties in achieving justice in trials. Researchers have been studying the sources and mechanisms of PCR inhibitors and developing mitigation strategies to facilitate DNA amplification in compromised samples.
FORENSIC SCIENCE INTERNATIONAL
(2023)
Article
Medicine, Research & Experimental
Francesco Nannini, Lenart Senicar, Farhaan Parekh, Khai J. Kong, Alexander Kinna, Reyisa Bughda, James Sillibourne, Xihao Hu, Biao Ma, Yuchen Bai, Mathieu Ferrari, Martin A. Pule, Shimobi C. Onuoha
Summary: Phage display technology combined with next-generation sequencing is a cutting-edge method for monoclonal antibody isolation. Utilizing third-generation sequencing, this study successfully identified functional scFv antibodies from diverse libraries against CD160 and CD123 that were previously undetected by conventional screening methods.
Article
Multidisciplinary Sciences
Giulia Agliardi, Anna Rita Liuzzi, Alastair Hotblack, Donatella De Feo, Nicolas Nunez, Cassandra L. Stowe, Ekaterina Friebel, Francesco Nannini, Lukas Rindlisbacher, Thomas A. Roberts, Rajiv Ramasawmy, Iwan P. Williams, Bernard M. Siow, Mark F. Lythgoe, Tammy L. Kalber, Sergio A. Quezada, Martin A. Pule, Sonia Tugues, Karin Straathof, Burkhard Becher
Summary: Research shows that local delivery of IL-12 combined with CAR-T cell therapy can achieve durable anti-tumor responses by enhancing cytotoxicity of CAR-T cells, reshaping the tumor microenvironment, promoting immune cell infiltration, and activating the myeloid compartment.
NATURE COMMUNICATIONS
(2021)
Letter
Oncology
P. A. Wawrzyniecka, L. Ibrahim, G. Gritti, M. A. Pule, P. M. Maciocia
Article
Immunology
Alexandre Klopp, Sophia Schreiber, Anna D. Kosinska, Martin Pule, Ulrike Protzer, Karin Wisskirchen
Summary: In this study, T cells were engineered to express HBV-specific receptors and inducible caspase 9 (iC9) as a safety switch, allowing for efficient recognition and killing of HBV-infected cells while also effectively depleting transferred T cells with unwanted hepatotoxicity both in vitro and in vivo.
FRONTIERS IN IMMUNOLOGY
(2021)
Article
Biochemical Research Methods
James E. Sillibourne, Giulia Agliardi, Matteo Righi, Katerina Smetanova, Grant Rowley, Simon Speller, Abigail Dolor, Katarina Lamb, Christopher Allen, Rajeev Karattil, Farhaan Parekh, Frederick Arce Vargas, Simon Thomas, Shaun Cordoba, Martin Pule
Summary: This study describes a simple method of achieving differential transgene expression by inserting stop codons and translational readthrough motifs (TRMs). TRMs can reduce downstream transgene expression and facilitate the controlled secretion of cytokines at therapeutically beneficial levels to prevent tumor growth. Due to their compact size and ease of introduction, TRMs are expected to be widely adopted in recombinant DNA engineering.
Article
Hematology
Paul M. Maciocia, Patrycja A. Wawrzyniecka, Nicola C. Maciocia, Amy Burley, Thaneswari Karpanasamy, Sam Devereaux, Malika Hoekx, David O'Connor, Theresa Leon, Tanya Rapoz-D'Silva, Rachael Pocock, Sunniyat Rahman, Giuseppe Gritti, Diana C. Yanez, Susan Ross, Tessa Crompton, Owen Williams, Lydia Lee, Martin A. Pule, Marc R. Mansour
Summary: CCR9 expression is found in the majority of T-ALL cases, making it a potential target for CAR-T cell therapy. CAR-T cells targeting CCR9 can effectively inhibit leukemia cell proliferation without harming healthy T cells, providing a promising treatment strategy for T-ALL.
Article
Hematology
Ilaria M. Michelozzi, Eduardo Gomez-Castaneda, Ruben V. C. Pohle, Ferran Cardoso Rodriguez, Jahangir Sufi, Pau Puigdevall Costa, Meera Subramaniyam, Efstratios Kirtsios, Ayad Eddaoudi, Si Wei Wu, Aleks Guvenel, Jonathan Fisher, Sara Ghorashian, Martin A. Pule, Christopher J. Tape, Sergi Castellano, Persis J. Amrolia, Alice Giustacchini
Summary: We have described a low-affinity second-generation CD19 chimeric antigen receptor (CAR) CAT that exhibits enhanced activation and cytokine polyfunctionality compared with high-affinity FMC63 CAR. This enhanced functionality is a result of antigen-dependent priming induced by residual CD19-expressing B cells. CAT CAR shows excellent toxicity profile, enhanced in vivo expansion, and long-term persistence in a phase 1 clinical study.
Article
Biotechnology & Applied Microbiology
Evangelia Kokalaki, Biao Ma, Mathieu Ferrari, Thomas Grothier, Warren Hazelton, Somayya Manzoor, Eren Costu, Julia Taylor, Anna Bulek, Saket Srivastava, Isaac Gannon, Ram Jha, Rosalind Gealy, Lukas Stanczuk, Tatiana Rizou, Mathew Robson, Mohamed El-Kholy, Vania Baldan, Matteo Righi, James Sillibourne, Simon Thomas, Shimobi Onuoha, Shaun Cordoba, Martin Pule
Summary: CART cells recognizing CD19 effectively treat relapsed and refractory B-ALL and DLBCL, but CD19 loss often leads to relapse. Targeting a second antigen, CD22, can reduce antigen escape, but it is challenging due to low CD22 density and large structure. The characteristics and optimal strategy for CD22 CAR are not well-studied. Co-administration of CD19 and CD22 CARs is costly, and constructing single CARs targeting both antigens is difficult. A dual CART product, CAT/9A8 CART, is being tested in a phase I clinical study as a potential solution.
Article
Medicine, Research & Experimental
Callum McKenzie, Mohamed El-Kholy, Farhaan Parekh, Mathew Robson, Katarina Lamb, Christopher Allen, James Sillibourne, Shaun Cordoba, Simon Thomas, Martin Pule
Summary: The hostile tumor microenvironment hampers the effectiveness of adoptive cell therapies. Disrupting the Fas death receptor, which initiates apoptosis, could enhance CAR T cell efficacy. By screening a library of Fas-TNFR proteins, researchers identified a novel chimera called Fas-CD40 that not only prevented Fas ligands from killing cells, but also synergistically enhanced CAR T cell efficacy.
MOLECULAR THERAPY-NUCLEIC ACIDS
(2023)
Article
Medicine, Research & Experimental
Leila Mekkaoui, Jose G. Tejerizo, Sara Abreu, Lydie Rubat, Aleksandra Nikoniuk, William Macmorland, Claire Horlock, Sofia Matsumoto, Sarah Williams, Koval Smith, Juliet Price, Saket Srivastava, Rehan Hussain, Mohammad Amin Banani, William Day, Elena Stevenson, Meghan Madigan, Jie Chen, Ravin Khinder, Shahed Miah, Simon Walker, Michael Ade-Onojobi, Sabine Domining, James Sillibourne, Marianna Sabatino, Vladimir Slepushkin, Farzin Farzaneh, Martin Pule
Summary: A simple one-step purification method using high-speed centrifugation was developed to purify transiently produced ?-RV for clinical application. The method achieved concentration of viral titers in the range of 10(7)-10(8) TU/mL with >80% overall recovery. Purification of ?-RV using this method resulted in significantly lower impurities compared to stable producer cell line vectors approved for clinical application.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2023)
Article
Biochemistry & Molecular Biology
Nathaniel D. Anderson, Jack Birch, Theo Accogli, Ignacio Criado, Eleonora Khabirova, Conor Parks, Yvette Wood, Matthew D. Young, Tarryn Porter, Rachel Richardson, Sarah J. Albon, Bilyana Popova, Andre Lopes, Robert Wynn, Rachael Hough, Satyen H. Gohil, Martin Pule, Persis J. Amrolia, Sam Behjati, Sara Ghorashian
Summary: In this study, the persistence of CD19 CAR-T cells in children with R/R B-ALL was analyzed, revealing a specific gene signature associated with long-lived CAR-T cells. This signature was found to be present in both children and adults with persistent CD19 CAR-T cells, indicating its potential as a universal transcriptional signature.
Article
Oncology
Ernesto Lopez, Rajesh Karattil, Francesco Nannini, Gordon Weng-Kit Cheung, Lilian Denzler, Felipe Galvez-Cancino, Sergio Quezada, Martin A. Pule
Summary: The combination of CAR T-cell therapy and MCT-1 blockade demonstrates improved cytotoxicity and antitumoral control in B-cell lymphoma, indicating the potential of targeting lactate metabolism in CAR T-cell therapies.
JOURNAL FOR IMMUNOTHERAPY OF CANCER
(2023)
Article
Oncology
Lydia Lee, Wen Chean Lim, Daria Galas-Filipowicz, Kent Fung, Julia Taylor, Dominic Patel, Zulaikha Akbar, Elena Alvarez Mediavilla, Patrycja Wawrzyniecka, Debarati Shome, Rogier M. Reijmers, Trillian Gregg, Leigh Wood, William Day, Virginie Cerec, Mathieu Ferrari, Simon Thomas, Shaun Cordoba, Shimobi Onuoha, Nushmia Khokhar, Vijay Peddareddigari, Muhammad Al-Hajj, Jim Cavet, Sonja Zweegman, Manuel Rodriguez-Justo, Kwee Yong, Martin Pule, Rakesh Popat
Summary: In this study, a ligand-based CAR targeting two myeloma antigens was constructed and evaluated in a clinical trial. The results showed that the APRIL CAR was well tolerated, but the clinical responses were disappointing. Comparing with other BCMA CARs, the APRIL CAR exhibited functional deficiencies due to reduced target binding by cell-expressed ligand.
JOURNAL FOR IMMUNOTHERAPY OF CANCER
(2023)
Article
Hematology
Ilaria M. Michelozzi, Eduardo Gomez-Castaneda, Ruben V. C. Pohle, Ferran Cardoso Rodriguez, Jahangir Sufi, Pau Puigdevall Costa, Meera Subramaniyam, Efstratios Kirtsios, Ayad Eddaoudi, Si Wei Wu, Aleks Guvenel, Jonathan Fisher, Sara Ghorashian, Martin A. Pule, Christopher J. Tape, Sergi Castellano, Persis J. Amroia, Alice Giustacchini
Summary: A low-affinity second-generation CD19 CAR showed improved expansion, cytotoxicity, and antitumor efficacy compared to a high-affinity CAR in preclinical models. In a phase 1 clinical study, the low-affinity CAR demonstrated an excellent toxicity profile and long-term persistence. In vitro characterization revealed that low-affinity CAR T cells had enhanced activation and a distinct transcriptomic and protein profile, with increased activation and cytokine polyfunctionality compared to high-affinity CAR T cells.
Article
Hematology
Claire Roddie, Lazaros J. Lekakis, Maria A. V. Marzolini, Aravind Ramakrishnan, Yiyun Zhang, Yanqing Hu, Vijay G. R. Peddareddigari, Nushmia Khokhar, Robert Chen, Silvia Basilico, Meera Raymond, Frederick Arce Vargas, Kevin Duffy, Wolfram Brugger, Maeve A. O'Reilly, Leigh Wood, David C. Linch, Karl S. Peggs, Carlos Bachier, Elizabeth Lihua Budde, Connie Lee Batlevi, Nancy Bartlett, David Irvine, Eleni Tholouli, Wendy Osborne, Kirit M. Ardeshna, Martin A. Pule
Summary: This study evaluated the efficacy of CD19/22 dual-targeting CAR-T (AUTO3) plus pembrolizumab in relapsed/refractory large B-cell lymphoma. The results showed that the AUTO3 plus pembrolizumab treatment regimen was safe for patients and provided durable remissions in complete responders.
