Synthetic biology has evolved from a set of engineering aspirations to an operationally sophisticated discipline, and artificial intelligence (AI) is its fastest-growing accelerant. This review traces that convergence across six interlocking domains: systems-level biological modeling, de novo protein engineering, metabolic and microbial programming, multi-omics data integration, regulatory element design, and clinical translation. For each domain, we survey established results, integrate findings from 2010–2026 literature, and articulate the trajectories that will define the next decade. Emerging themes include physics-informed neural networks for mechanistically constrained biological modeling, drug design, and federated learning architectures that allow global omics collaboration without centralizing sensitive data, self-driving laboratories that close the Design-Build-Test-Learn loop with minimal human intervention, and large language models that accelerate hypothesis generation from scientific literature. Alongside these opportunities, the review gives equal weight to the governance challenges they create: dual-use risks amplified by generative sequence design, the reproducibility crisis in AI-driven biodesign, and the equitable distribution of autonomous experimentation capacity. The overarching argument is that the promise of synthetic biology, making biological design as deliberate and reliable as any mature engineering discipline, is closer than ever, but will only be realized if technical ambition is matched by scientific rigor, transparent governance, and inclusive access.

Therapeutic monoclonal antibodies can elicit anti-drug antibodies (ADAs) in patients, which impact drug pharmacokinetics, reduce efficacy, and occasionally cause hypersensitivity reactions. Growing evidence indicates that early detection of ADA responses improves therapy outcomes, yet their monitoring has remained underappreciated. Conventional assays are complex, time-consuming, and confined to centralized laboratories, thereby delaying clinical decisions. In contrast, emerging biosensor technologies represent a promising alternative for timely, on-site ADA detection. This review examines advancements in ADA detection and provides a critical overview of its clinical relevance based on contemporary findings. Furthermore, the challenges and complexities of accurately biosensing ADAs are discussed. Finally, we explore the prospects of biosensors for on-site analysis and their integration into proactive therapeutic drug monitoring.

Wildfires that destroy residential infrastructure can generate chemically complex soil contamination; however, post-fire screening is often limited and does not directly reflect biological hazards. Herein, we integrated a multi-stress lux-based whole-cell bioreporter panel of genetically engineered Escherichia coli strains with non-targeted LC-MS profiling to obtain a mechanism-informed assessment of soils collected from a residential property impacted by the January 2025 Los Angeles wildfires. The bioreporter panel resolved heterogeneous and statistically significant stress signatures across the analyzed samples. In particular, extracts from U3-U5 produced selective suppression of the membrane and fatty acid biosynthesis bioreporters, along with reduced growth. In contrast, extract U5 induced a proteotoxic heat-shock response signature. In parallel, non-targeted LC-MS detected 1813 chemical features and enabled the putative annotation of a subset of signals consistent with combustion-derived organics and reactive electrophiles, providing a chemical context for the observed bioassay fingerprints. The integrated workflow provides mechanism-resolved hazard triage within 48 h, as implemented herein (24 h elutriate preparation plus up to 20 h microplate kinetics), supporting the prioritization of hotspots for confirmatory analysis, remediation, and risk assessment.

The transmission history and adaptive evolution of Mycobacterium tuberculosis complex (MTBC) in China remain underexplored despite its remarkably low diversity and enduring public health burden. Here, we analyzed 23,873 whole genome sequences of MTBC to reconstruct its spread timeline and routes, population dynamics, and host adaptation patterns in China. The Bayesian coalescent models revealed the recurrent introductions of four MTBC sub-lineages (L2.2, L4.2, L4.4, and L4.5) during 1000-400 years ago, by European-Chinese transmission networks, which might have triggered the formation of their local genetic clades in China. These local clades underwent three rapid population expansions that temporally aligned with historical climate cooling events and warfare, and displayed convergent adaptation, including shared mutations and structural variations in macrophage-resistance genes and enhanced genetic diversity in T cell epitopes. The historical mutation rate estimations and neutral mutation simulations indicated that these local clades experienced pronounced macrophage-induced pressures, likely operative over the past two centuries. The DNA information entropy analysis further revealed their adaptive evolution signatures clustered within macrophage-resistance pathways. The gene Rv0801, computationally predicted to exhibit the most prominent adaptive signatures despite its uncharacterized function, was confirmed through recombinant strain infection to enhance intracellular survival in human macrophages. This integrative approach reveals MTBC's evolutionary trajectory in China, provides novel methods for quantifying selection pressures and detecting adaptive evolution signals in prokaryotes, and constructs a comprehensive framework for investigating pathogen transmission dynamics and host adaptation mechanisms.

β-lactams are an important family of antibiotics that are prone to undergo resistance inhibition though the production of β-lactamases by some microorganisms. To combat this resistance and preserve the efficacy of β-lactam antibiotics, we developed a strategy for the discovery of such β-lactamase inhibitors. When combined with β-lactams, these inhibitors allow the antibiotics to be effective and prevent resistance. To date, the development of such combinatory drugs is limited due to the complexity of screening for new β-lactamase inhibitors. Therefore, to facilitate this development, it was essential to find sensitive assays to effectively screen for lactamase inhibitory compounds. To this end, a novel bioassay utilizing bioluminescent indicator bacteria as bioreporters was developed. The assay was first optimized using commercial antibiotics together with known β-lactamase inhibitors. Using this bioassay, we then screened for novel natural β-lactamase inhibitors derived from coral-associated fungi. We showed that the fungus Penicillium spinulosum, originating from the coral Pocillopora sp. from the Gulf of Aqaba Eilat, produced compounds with anti-β-lactamase activity. We further demonstrated that the bioreporter bacteria used here responded to the combined antibiotics and β-lactamase inhibitors in a concentration-dependent manner, indicating their usefulness for β-lactamase-inhibiting compound discovery. Future structural identification will promote the validation of this assay’s usefulness.

Administration of mAb therapeutics often elicits the production of neutralizing anti-drug antibodies (nADA) in patients. Current nADA assays are drug-sensitive and detect only free nADA, thereby overlooking drug-bound antibodies and reducing sensitivity. An extra preparatory step of acid dissociation is often applied to resolve the immunocomplexes and subsequently detect the resultant free nADA separately. This tedious sample preparation limits them to lab settings and exposes the antibodies to harsh conditions. Here, a modified lateral flow assay platform, namely sequential binding flow immunoassay (SBFIA) is presented for nADA determination that circumvents the drug's interference through a distinctive approach. Drawing inspiration from chromatography, sequential binding events (competitive and direct) are exploited to eliminate drugs' interference, enabling sensitive detection of drug-bound nADA. The platform is optimized using Infliximab (IFX) as a drug and neutralizing anti-Infliximab antibodies (n alpha IFX) as a model analyte. After optimization, a minimum concentration of 103 ng mL(-1) n alpha IFX is detected. The assay shows remarkable drug tolerance and specificity to n alpha IFX. Based on its point-of-care (POC) advantages while avoiding harsh, acidic sample pretreatment, the assay can assist physicians in rapid and accurate decision-making. By tweaking the bioreceptors, the configuration can adapt well to other biologics and respective nADA.

Hybridoma technology remains a cornerstone of monoclonal antibody (mAb) discovery. Classical screening practices such as ELISA, western blot, and dot blot require laborious, time-consuming procedures, rendering them inefficient in time-restricted decision-making. Additionally, due to these assays' practical and technical limitations, specificity testing of the mAbs is usually omitted during the primary screening of hybridoma libraries. Herein, we present a rapid, dipstick immunoassay (DIA) designed for mAbs screening in cell culture supernatant. The integrated proprietary setup is based on antibody capture and comprises accessible materials such as conjugate pad, nitrocellulose membrane, and absorbent pad. The critical element of this technology is the design of the assay. During the assay run, dipsticks are inserted into supernatant-containing microtiter wells, initiating capillary flow of mAbs towards the conjugate pad where a pre-dried HRP-labeled anti-host species antibody is embedded. This interaction forms an immunocomplex, which migrates to the nitrocellulose membrane and binds to the pre-immobilized target antigen while simultaneously encountering immobilized putative cross-reacting proteins in a multiplex-line fashion. Upon addition of HRP-oxidizable substrate, signal acquisition is performed using a simple camera (colorimetry) or a CCD sensor (chemiluminescence). The system was quantitative up to 2500 ng mL- 1 and showed a minimum detectable concentration of 0.61 ng mL- 1. Compared to the gold-standard ELISA, the sensitivity was 8-fold higher, while the dynamic range was similar. Critically, the assay allows for the concurrent assessment of antibody specificity in one run. This immunoassay's unique configuration, simplicity, and rapidity can facilitate antibody discovery.

Cigarette smoking is known to be an unhealthy activity that can cause a number of human diseases, including chronic obstructive pulmonary disease (COPD) and lung cancer. It was further reported that even being exposed to secondhand cigarette smoke can affect human health. To assess the toxicity of the smoke from different cigarette brands, an artificial smoking device was developed, and three fractions designated, Filter Fraction, Smoke Fraction and Tar Fraction, were prepared from the smoke of each brand. Then, to elucidate possible effects of some of the toxins found in cigarette smoke, we investigated their effects in vitro using a bioluminescent bacterial array that comprises three bacterial strains. Using this array, we compare smoke from three cigarette brands, each with different tar and nicotine contents. GC-MS analysis showed that the cigarette smoke extracts (fractions) from different brands differed in their compositions and chemical concentrations. The results further showed that, in general, cigarette smoke triggered mainly an oxidative stress reaction in our bacterial models. The Smoke Fraction was tested for sequential smoking rounds and found to produce cumulative effects following each subsequent smoking cycle for all three cigarette brands. Finally, it was found that cigarette smoke and its specific components are toxic at various degrees with the Smoke Fraction, acting as oxidative stressors, and that this can be effectively analyzed using bioreporter panel arrays.

Electrically conductive hydrogels are gaining attention owing to their applications in biosensing, cellular interfaces, and tissue engineering. However, conventional hydrogels often lack adequate electrical conductivities. Here, we present two novel conductive alginate-based hydrogels designed for extrusion-based 3D bioprinting: (i) covalently synthesized alginate–polypyrrole (alginate–PPy) via EDC/NHS-mediated conjugation with 3-aminopropyl pyrrole, and (ii) nanoparticle-reinforced alginate blended with polypyrrole nanoparticles (alginate@PPy-NP). Both systems exhibited shear-thinning behavior, tunable viscoelasticity, and excellent printability. Alginate@PPy-NP demonstrated superior compressive strength and shape fidelity, whereas alginate–PPy showed enhanced elastic moduli (G′/G″), reflecting a more uniform gel network. Electrical conductivity increased with increasing pyrrole content in both formulations. Optimization of the composition and printing conditions enabled the fabrication of fibroblast-laden constructs with high structural integrity. This work highlights the potential of alginate–polypyrrole hydrogels as customizable, conductive bioinks for 3D bioprinting in regenerative medicine.

Analytical instruments used for scientific research are often expensive. This limitation poses a challenge for researchers in low-income laboratories. To address this problem, the improvization of analytical instruments has emerged as a viable solution. Herein, we review and discuss, step-by-step, the approach of using a smartphone for colorimetric analysis, known as smartphone digital image colorimetry (SDIC). With its practicality and accessibility, SDIC utilizes smartphone digital cameras for image acquisition and free image processing programs, and smartphone Apps, for analysis. By leveraging the ubiquity and affordability of smartphone technology, SDIC offers an accessible and cost-effective approach to researchers in resource-constrained settings.

Sediments are key players in the optimum functioning of ecosystems; however, they also represent the largest known repository of harmful contaminants. The vast variety of these sediment-associated contaminants may exert harmful effects on marine communities and can impair ecosystem functioning. Whole-cell biosensors are a rapid and biologically relevant tool for assessing environmental toxicity. Therefore, in this study, we developed a bioassay-based toxicity measurement system using genetically modified bacteria to create a whole-cell optical biosensor. Briefly, reporter bacteria were integrated and immobilized using a calcium alginate matrix on fiber-optic tips connected to a photon counter placed inside a light-proof, portable case. The calcium alginate matrix acts as a semi-permeable membrane that protects the reporter-encapsulated optical fiber tips and allows the inward passage of toxicant(s) to induce a dose-dependent response in the bioreporter. The samples were tested by directly submerging the fiber tip with immobilized bacteria into vials containing either water or suspended sediment samples, and the subsequent bioluminescent responses were acquired. In addition to bioavailable sediment toxicity assessments, conventional chemical methods, such as liquid chromatography–mass spectroscopy (LC-MS) and inductively coupled plasma optical emission spectroscopy (ICP-OES), were used for comprehensive evaluation. The results demonstrated the efficacy of the biosensor in detecting various toxicity levels corresponding to identified contaminants, highlighting its potential integration into environmental monitoring frameworks for enhanced sediment and water quality assessments. Despite its utility, this study notes the system’s operational challenges in field conditions, recommending future enhancements for improved portability and usability in remote locations.

Biofilms cling to surfaces to form complex architectures allowing their bacterial creators to acquire multidrug resistance and claiming countless lives worldwide. Therefore, finding novel compounds that affect virulence and biofilm-forming capacity of resistant pathogenic bacteria is imperative. Recently, we identified indole-based compounds that possess anti-biofilm properties in coral-associated bacteria. We succeeded in efficiently synthesizing two of these compounds, 1,1’-bisindole (NN) and 2,3-dihydro-2,2’-bisindole (DIV). They were found to attenuate biofilms of gram-negative bacterial pathogens, including Pseudomonas aeruginosa and Acinetobacter baumannii. Combining these compounds with the antibiotic tobramycin resulted in significant biofilm inhibition, particularly in the eradication of mature P. aeruginosa biofilms. Both of the bisindole derivatives, suppressed a number of bacterial virulence factors, reduced bacterial adhesion, and improved survival rates in infected Caenorhabditis elegans and human lung epithelial cell models. Transcriptome analyses of the bacteria treated with these compounds revealed that NN repressed or upregulated 307 genes when compared to untreated P. aeruginosa. These bacteria-derived molecules act in resistance-quenching and are potentially important candidates for inclusion in treatment protocols. The use of compounds that prevent the biofilm from accumulating the high cell densities critical to its structural and functional maintenance represents significant progress in the management of bacterial persistence. Therefore, a possible clinical implementation of these innovative compounds holds a promising future.

Metastatic cervical lymph nodes (LN) are detected in 20-30% of patients with differentiated thyroid cancer (DTC). Current guidelines recommend that once a cervical LN is suspected to be DTC metastasis during a neck ultrasound (US) procedure, it should be investigated via a fine needle aspiration (FNA) biopsy for cytological evaluation and saline washout of the needle for thyroglobulin (Tg) measurement (FNA-Tg). Since Tg is a protein produced exclusively by thyroid follicular cells, a positive FNA-Tg result establishes the diagnosis of metastatic DTC irrespective of cytology. The conventional, immunoassay-based, FNA-Tg washout requires a laboratory and skilled personnel. We developed a semi-quantitative, lateral flow-based method which was shown to detect at the point-of-care (POC), within 10 minutes, positive Tg samples in needle washouts of a suspicious LN at the site of FNA biopsy. In the pre-clinical phase, the POC-Tg limit of detection was determined to be at a concentration equal to 5 ng/mL, after a 1 mL dilution with normal saline. Our prototype was optimized by evaluating different components: types of membranes, pads, antibodies, and gold conjugates. We evaluated our POC-Tg kits on thirty clinical samples: 16 were found positive while the other 14 were seen as negative. All the negative and positive results were further validated by the attending clinical labs, resulting in 100% compatibility compared to the standard procedure. The proof-of-value of our POC-Tg test lies in its ability to significantly reduce the time to results, thus enhancing clinical decision-making, and saving time and valuable resources. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This study was funded by the Israel Innovation Authority [Kmin grant, number 64994], and the Israel Cancer Association [grant number 20230021]. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethical approval for this study was obtained by the Helsinki committee of Soroka University Medical Center (SUMC), approval number 190-17-SOR. All patients provided written informed consent before the FNA procedure was initiated. Patients 18-year-old and older, able to understand and sign the informed consent form, who were evaluated for cervical LN suspected as DTC metastases, were offered to participate. Pregnant women were excluded. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors

The present study offers a new method for analyzing the bioluminescence kinetics generated by the toxicity-triggered bioluminescent bacterial mutant strain TV1061 to detect and differentiate heavy metals in water samples. Specifically, as a proof-of-concept, copper and mercury were used, in single and binary mixtures, to induce the grpE heatshock promoter generating a measurable signal. The research employed the Dynamic Time Warping (DTW) algorithm to analyze the overall bioluminescence signal generated by the bacteria, rather than focusing on specific signal components, such as the response ratio. The method demonstrated high accuracy in classifying the samples (94 % accuracy). This technique is a first stepping stone in creating a database that will enable to accuractly identify mixtures of toxicants and predict their concentration in the sample. This approach is reliable, cost-effective, and rapid for monitoring and assessing toxic contaminants in water, including deciphering distinct recognition patterns, such as for copper and mercury (our chosen case specimens), including in binary mixtures, highlighting its potential for the precise identification and quantification of heavy metals in complex mixtures. Our findings support the possibility of developing a larger dataset of multiple references for multiple heavy metals and other toxicants, that will improve the overall detection capabilities of our system, all this thanks to the combined use of data mining and machine learning techniques.

A novel porous metal-organic framework (MOF), pCu-BDC-NH2, with hierarchical porosity was synthesized using cetyltrimethylammonium bromide (CTAB) as a pore-generation agent. In addition to its common functions including structure-directing ligands or soft micelle templates, the judicious use of CTAB effectively modulated pore architecture in Cu-BDC-NH2 MOFs. With additional mesopores generated during the synthesis process, the intrinsic MOF scaffolds further obtained pore hierarchies and interconnectivity, enabling efficient substrate access to the active metal centers, and thus significantly facilitated catalytic performance. As a proof of concept, we applied the finely engineered porous MOF pCu-BDC-NH2 in a cascaded enzymatic system for xanthine sensing. This colorimetric biosensor exhibited a low detection limit of 0.11 μM, and a wide linear range of 1–120 μM. Furthermore, the sensor demonstrated exceptional stability, reproducibility, and was independent of interferences. Our simple yet effective method may find broader applications in tailoring pore architecture, enabling finer engineered structures to improve catalytic activities of nanomaterials.

Liver congestion is increasingly encountered in clinical practice and presents diagnostic pitfalls of which radiologists must be aware. The complex altered hemodynamics associated with liver congestion leads to diffuse parenchymal changes and the development of benign and malignant nodules. Distinguishing commonly encountered benign hypervascular lesions, such as focal nodular hyperplasia (FNH)-like nodules, from hepatocellular carcinoma (HCC) can be challenging due to overlapping imaging features. FNH-like lesions enhance during the hepatic arterial phase and remain isoenhancing relative to the background liver parenchyma but infrequently appear to wash out at delayed phase imaging, similar to what might be seen with HCC. Heterogeneity, presence of an enhancing capsule, washout during the portal venous phase, intermediate signal intensity at T2-weighted imaging, restricted diffusion, and lack of uptake at hepatobiliary phase imaging point toward the diagnosis of HCC, although these features are not sensitive individually. It is important to emphasize that the Liver Imaging Reporting and Data System (LI-RADS) algorithm cannot be applied in congested livers since major LI-RADS features lack specificity in distinguishing HCC from benign hypervascular lesions in this population. Also, the morphologic changes and increased liver stiffness caused by congestion make the imaging diagnosis of cirrhosis difficult. The authors discuss the complex liver macro- and microhemodynamics underlying liver congestion; propose a more inclusive approach to and conceptualization of liver congestion; describe the pathophysiology of liver congestion, hepatocellular injury, and the development of benign and malignant nodules; review the imaging findings and mimics of liver congestion and hypervascular lesions; and present a diagnostic algorithm for approaching hypervascular liver lesions.

Three-dimensional (3D) printing technology, also known as additive manufacturing (AM), has emerged as an attractive state-of-the-art tool for precisely fabricating functional materials with complex geometries, championing several advancements in tissue engineering, regenerative medicine, and therapeutics. However, this technology has an untapped potential for biotechnological applications, such as sensor and biosensor development. By exploring these avenues, the scope of 3D printing technology can be expanded and pave the way for groundbreaking innovations in the biotechnology field. Indeed, new printing materials and printers would offer new possibilities for seamlessly incorporating biological functionalities within the growing 3D scaffolds. Herein, we review the additive manufacturing applications in biosensor technologies with a particular emphasis on extrusion-based 3D printing modalities. We highlight the application of natural, synthetic, and composite biomaterials as 3D-printed soft hydrogels. Emphasis is placed on the approach by which the sensing molecules are introduced during the fabrication process. Finally, future perspectives are provided.

Selective and sensitive detection of nitrite has important medical and biological implications. In the present work, to obtain an enhanced electrochemiluminescence (ECL) determination of nitrite, a novel nano-ECL emitter CoBIM/cetyltrimethylammonium bromide (CTAB) was prepared via a micelle-assisted, energy-saving, and ecofriendly method based on benzimidazole (BIM) and CTAB. Unlike conventional micelle assistance, the deprotonated BIM (BIM-) preferential placement was in the palisade layer of cationic CTAB-based micelles. Enriching the original CTAB micelle with BIM- disrupted its stability and resulted in the formation of considerably smaller BIM/CTAB-based micelles, providing a confined coordination environment for BIM- and Co2+. As a result, the growth of CoBIM/CTAB was also limited. Owing to the unusual nitration reaction between BIM and nitrite, the prepared CoBIM/CTAB was successfully applied as a novel ECL probe for the detection of nitrite with a wide linear range of 1-1500 mu M and a low detection limit of 0.67 mu M. This work also provides a promising ECL platform for ultrasensitive monitoring of nitrite and it was applied with sausages and pickled vegetables.

The aim of this study is to prospectively evaluate whether women with copper-containing intrauterine devices (Cu-IUD), currently listed as MR conditional, can safely undergo 3.0 Tesla (3 T) magnetic resonance imaging (MRI). 73 women, age 18–54 years old, with a Cu-IUD who were undergoing MRI for any reason were included consecutively. Pre- and post-MRI standard pelvic ultrasound examinations were completed to determine the appropriate pre- and post-MRI positioning of the Cu-IUD. Displaced IUDs were defined by IUD crossbars not in the fundal portion of the endometrial cavity, a visualized tip in the mid or lower uterus, any part of the device located in the cervical canal or outside of the endometrial canal, a fractured device, or a non-visualized IUD. Additionally, a questionnaire was completed by participants to determine the level of pre- and post-MRI pelvic pain. There were zero observed displaced Cu-IUDs on post-MRI pelvic ultrasounds (p = 0/70, 95

The design of Prussian blue nanoparticles (PB NPs) with satisfied electrocatalytic ability and stability is still challenging. Herein, a hierarchical confinement strategy of PB NPs is proposed using NH2MIL-88B(Fe) (denoted herein as NH2MIL) as both self-confined template and Fe3+ sources, K-3[Fe(CN)(3)] as precursor. The catalytic synthesis of PB NPs is further achieved during the hydrothermal process at 100 C-degrees for 1 h. The as-prepared PB@NH2MIL exhibits the morphology of "sea cucumber", retaining the intrinsic skeleton of NH2MIL dotted with lots of PB NPs (<40 nm). In addition, the initial coordination of [Fe(CN)(3)](3-) with the active free coordination sites of NH2MIL results in the coatom phenomenon between NH2MIL and PB NPs. Thus, the as-prepared PB@NH2MIL possesses the efficient electronic transmission, the enhanced electrocatalytic properties as well as stability under prolonged electrochemical measurements. Finally, the as-prepared PB@NH2MIL can be used as artificial peroxidase for the electrochemical detection of H2O2 with a wider linear range (0.01-1000 mu M) and low detection limit (3 nM).