Screening Step Research Articles

A sequential explanatory mixed method study of maternal and fetal outcome in gestational diabetes mellitus using Diabetes in Pregnancy Study Group India (DIPSI) test in Puducherry

ABSTRACT Background: Gestational diabetes mellitus in pregnancy is associated with polyhydramnios, macrosomia, and shoulder dystocia, and it also increases maternal and perinatal mortality. Methods: This sequential explanatory mixed-method study was conducted for six months. All the pregnant women attending the outpatient department of the Obstetrics and Gynaecology Department at 24-28 weeks of gestation were subjected to universal screening with 75 gms of glucose and 2 hours of plasma glucose >140 mgs% is taken for diagnosis (according to DIPSI guidelines). After diagnosis, they were subjected to an HbA1c test. Women with HbA1c is >6.5% were excluded from the study. If pregnant women are screened negative by the DIPSI test, the test was repeated in the third trimester (32-34 weeks of gestation). Chi-square tests were used to find out the test of association for quantitative data and manual content analysis was performed for qualitative data. Results: DIPSI test was found to decrease the adverse maternal and neonatal outcome by early screening and management. The stakeholders’ perspectives identified by key informant interview were improper knowledge and awareness about the testing and others were anxiety and fear associated with the testing procedure. Conclusions: As DIPSI test is an effective single step in screening and diagnostic test, hence all pregnant mothers should undergo this glucose challenge test in their antenatal visits.

RRBM-YOLO: Research on Efficient and Lightweight Convolutional Neural Networks for Underground Coal Gangue Identification.

Coal gangue identification is the primary step in coal flow initial screening, which mainly faces problems such as low identification efficiency, complex algorithms, and high hardware requirements. In response to the above, this article proposes a new "hardware friendly" coal gangue image recognition algorithm, RRBM-YOLO, which is combined with dark light enhancement. Specifically, coal gangue image samples were customized in two scenarios: normal lighting and simulated underground lighting with poor lighting conditions. The images were preprocessed using the dim light enhancement algorithm Retinexformer, with YOLOv8 as the backbone network. The lightweight module RepGhost, the repeated weighted bi-directional feature extraction module BiFPN, and the multi-dimensional attention mechanism MCA were integrated, and different datasets were replaced to enhance the adaptability of the model and improve its generalization ability. The findings from the experiment indicate that the precision of the proposed model is as high as 0.988, the mAP@0.5(%) value and mAP@0.5:0.95(%) values increased by 10.49% and 36.62% compared to the original YOLOv8 model, and the inference speed reached 8.1GFLOPS. This indicates that RRBM-YOLO can attain an optimal equilibrium between detection precision and inference velocity, with excellent accuracy, robustness, and industrial application potential.

Multimodal human-computer interaction in interventional radiology and surgery: a systematic literature review.

As technology advances, more research dedicated to medical interactive systems emphasizes the integration of touchless and multimodal interaction (MMI). Particularly in surgical and interventional settings, this approach is advantageous because it maintains sterility and promotes a natural interaction. Past reviews have focused on investigating MMI in terms of technology and interaction with robots. However, none has put particular emphasis on analyzing these kind of interactions for surgical and interventional scenarios. Two databases were included in the query to search for relevant publications within the past 10 years. After identification, two screening steps followed which included eligibility criteria. A forward/backward search was added to identify more relevant publications. The analysis incorporated the clustering of references in terms of addressed medical field, input and output modalities, and challenges regarding the development and evaluation. A sample of 31 references was obtained (16 journal articles, 15 conference papers). MMI was predominantly developed for laparoscopy and radiology and interaction with image viewers. The majority implemented two input modalities, with voice-hand interaction being the most common combination-voice for discrete and hand for continuous navigation tasks. The application of gaze, body, and facial control is minimal, primarily because of ergonomic concerns. Feedback was included in 81% publications, of which visual cues were most often applied. This work systematically reviews MMI for surgical and interventional scenarios over the past decade. In future research endeavors, we propose an enhanced focus on conducting in-depth analyses of the considered use cases and the application of standardized evaluation methods. Moreover, insights from various sectors, including but not limited to the gaming sector, should be exploited.

Analyzing the existing programs on promoting women scientists in chemistry

Abstract Gender imbalance in science has been well documented. Possible explanations include a lack of systematic strategies or policies to support women in science in general and chemistry in particular. This article aims to analyze selected programs that advocate gender equity and inclusion in chemistry. Forty-eight initiatives were selected via several steps of screening. Following a pilot study using systematic review as a research method, the named initiatives were analyzed considering their strategies, goals, and impact. The findings reveal that acknowledging women scientists and providing funding to enable and sustain networking (such as through conferences) are crucial for women scientists to become visible and recognized in academia. In this paper, we offer recommendations for other entities who would like to develop initiatives to encourage suitable environments for gender equity.

Engaging Diverse African American/Black and Latine Youth and Emerging Adults Living with HIV into Research: Description of Recruitment Strategies and Lessons Learned.

Improving engagement along the HIV care continuum and reducing racial/ethnic disparities are necessary to end the HIV epidemic. Research on African American/Black and Latine (AABL) younger people living with HIV (LWH) is essential to this goal. However, a number of key subgroups are challenging to locate and engage, and are therefore under-represented in research. Primary among these are persons with non-suppressed HIV viral load, severe socioeconomic disadvantage, transgender/gender expansive identities, and refugee/migrant/immigrant populations. Research in community settings is needed to complement studies conducted in medical institutions. The present study describes the efficiency of recruitment strategies used in the community to enroll AABL young and emerging adults LWH ages 19-28 years. Strategies were designed to be culturally responsive and structurally salient. They were: peer-to-peer, social media, classified advertisements (newspaper, craigslist), subway ads, dating apps (Jack'd, Positive Singles), and direct recruitment in community-based organizations. Data were analyzed using mainly descriptive statistics and interpreted using a consensus building approach. We screened 575 individuals in a first step, 409 were eligible (71%), of these 297 presented to the second screening step (73%), but 112 were lost. Almost all presenting at the second step were eligible (98%, 291/297) and 94% enrolled (274/291). Peer-to-peer, dating app (Jack'd), direct recruitment, and craigslist were the most efficient strategies. Recruitment on dating apps was superior to the peer-to-peer approach in yielding eligible participants (OR = 1.5; 95% CI: 0.98-2.3; p = 0.06). The sample enrolled was diverse with respect to HIV viral suppression, gender identify, sexual orientation, immigration status, and barriers to HIV care engagement. We discuss the advantages and disadvantages of each strategy. Recruitment is a vital aspect of research and warrants attention in the empirical literature.

Investigating Full-Length circRNA Transcripts to Reveal circRNAMediated Regulation of Competing Endogenous RNAs in Gastric Cancer

Background: Circular RNAs (circRNAs) play important regulatory roles in the progression of gastric cancer (GC), but the exact mechanisms governing their regulation remain incompletely understood. Prior studies typically used back-spliced junctions (BSJs) to represent a range of circRNA isoforms, overlooking the prevalence of alternative splicing (AS) events within circRNAs, which could lead to unreliable or even incorrect conclusions in subsequent analyses, hindering our comprehension of the specific functions of circRNAs in GC. Objective: This study aimed to explore the potential functional roles of the dysregulated circRNA transcripts in GC and provide new biomarkers and effective novel therapeutic strategies for GC treatment. Methods: RNA-seq data with rRNA depletion and RNase R treatment was employed to characterize the expression profiles of circRNAs in GC, and RNA-seq data only with rRNA depletion was employed to identify differentially expressed mRNAs in GC. Based on the full-sequence information and accurate isoform-level quantification of circRNA transcripts calculated by the CircAST tool, we performed a series of bioinformatic analyses. A circRNA-miRNA-hub gene regulatory network was constructed to reveal the circRNA-mediated regulation of competing endogenous RNAs in GC, and then the protein-protein interaction (PPI) network was built to identify hub genes. Results: A total of 18,398 circular transcripts were successfully reconstructed in the samples. Herein, 351 upregulated and 177 downregulated circRNA transcripts were identified. Functional enrichment analysis revealed that their parental genes were strongly associated with GC. After several screening steps, 19 dysregulated circRNA transcripts, 40 related miRNAs, and 65 target genes (mRNAs) were selected to construct the ceRNA network. Through PPI analysis, five hub genes (COL5A2, PDGFRB, SPARC, COL1A2, and COL4A1) were excavated. All these hub genes may play vital roles in gastric cancer cell proliferation and invasion. Conclusion: Our study revealed a comprehensive profile of full-length circRNA transcripts in GC, which could provide potential prognostic biomarkers and targets for GC treatment. The results would be helpful for further studies on the biological roles of circRNAs in GC and offer new mechanistic insights into the pathogenesis of GC.

The Sansure® Human Papillomavirus DNA Diagnostic Kit offers excellent reproducibility performance for the detection of high-risk HPV.

Cervical cancer screening is a cornerstone of cervical cancer elimination. Detection of high-risk human papillomavirus (hrHPV) is recommended as the first step in screening provided that the assay used has been adequately validated. The Sansure® Human Papillomavirus DNA Diagnostic Kit is a new assay designed to detect HPV16, HPV18 and 13 other HPV in aggregate. The study aimed to evaluate the intra- and interlaboratory reproducibility of the assay according to international guidelines. Five hundred and fifty cervical residual cell samples from women attending cervical cancer screening were selected from the biobank of the HPV National Reference Centre in Belgium and used in this study. After DNA extraction, HPV was tested using the Sansure® Human Papillomavirus DNA Diagnostic Kit. The lower 95% confidence limit around the general reproducibility of this assay should be greater than or equal to 87%, with κ ≥ 0.50. Five hundred and thirty-three samples had valid results. The Sansure® Human Papillomavirus DNA Diagnostic Kit demonstrated an excellent intra-laboratory reproducibility of 93.8% (95% confidence interval [CI]: 91.4-95.7, κ = 0.85). The interlaboratory reproducibility was 93.4 (95% CI: 91.0-95.4, κ = 0.84). Intra and interlaboratory reproducibility were also excellent at the genotype level. Excluding HPV53 single infection samples from the analyses also resulted in excellent agreement. These data show that the Sansure® Human Papillomavirus DNA Diagnostic Kit is highly reproducible.

Parental involvement in students’ mathematics activities: A bibliometric analysis

This study is focused on investigating parental involvement in students’ mathematical activities. It presents a comprehensive bibliometric analysis that examines new areas of research to discover current trends and future research opportunities related to parental involvement in their children’s mathematics activities. This study intends to provide a meaningful contribution toward enhancing educational interventions in the future. The metadata of the publications included in this analysis was extracted from the Scopus and ERIC databases using the search criteria “(parent OR parent) AND (involvement OR support) AND (math or math) AND student.” After the screening step, a comprehensive analysis was conducted on a total of 944 articles using the tools Harzing’s Publish or Perish 8, Biblioshiny, and VOSviewer 1.6.20. The findings indicated that studies on this topic covered 42 years and accumulated a total of 17,392 citations. The most prolific year in terms of publications was 2023, with a peak of 80 studies published. The leading contributors to this body of research were Jianzhong Xu and James Reed Campbell, each authoring six papers. The keyword mapping results revealed several interesting research avenues for future investigation, such as the incorporation of psychological interventions, longitudinal studies, multidisciplinary approaches, and the utilization of technology.

Pharmacy stakeholders' views and experiences of the credentialing of advanced or specialist pharmacist practice: A mixed methods systematic review

BackgroundCredentialing of advanced and specialist pharmacist practice (ASPP) provides essential quality assurance for ASPP, which is significantly different to entry-level practice and is developing worldwide. Several credentialing models are in place or under development internationally. Synthesis of the views and experiences of pharmacists and other relevant stakeholders on credentialing is an important research gap. Objective(s)To determine pharmacy stakeholders' views and experiences of ASPP credentialing and to explore facilitators and barriers to credentialing implementation and uptake. MethodsThe Joanna Briggs Institute convergent integrated approach for mixed methods systematic reviews was followed. The review protocol was pre-registered with PROSPERO. Five electronic databases (Medline, CINAHL, Embase, Web of Science, Google Scholar) were searched from inception to August 2022 for qualitative, quantitative, or mixed methods studies that reported the views and experiences of pharmacy stakeholders on ASPP credentialing. The Mixed Methods Appraisal Tool was used to quality appraise included studies. Thematic synthesis was undertaken to analyse and integrate data from included studies. All screening and data analysis steps were performed by two reviewers independently, with additional author input where required. ResultsSixty studies were included, after screening titles and abstracts (n = 9055) and full texts (n = 228). Studies represented the views of pharmacists, pharmacy managers/employers and professional/representative bodies in hospital, community pharmacy, general practice/primary care, and academic sectors, from 40 countries. Four analytical themes were generated describing the factors, including facilitators and barriers, to be considered when developing or optimising ASPP credentialing: I. Drivers of credentialing, II. Developing ASPP competence, III. Optimising credentialing implementation, and IV. Enhancing credentialing uptake. ConclusionsThis systematic review is the first to synthesise pharmacy stakeholders' views and experiences of ASPP credentialing. A conceptual framework highlights contextual factors, facilitators, barriers, and inter-relationships which should be considered by pharmacists, policymakers, and other key stakeholders when implementing ASPP credentialing.

Pre-marital HIV screening in Jayapura Regency: A phenomenological study

Background: Premarital screening is a series of examinations that prospective bridal couples must undergo. Infectious disease screening is one of the important screening steps that prospective brides and grooms must carry out, including examination for hepatitis B, hepatitis C, sexually transmitted infections (STIs), and HIV-AIDS. Premarital HIV screening is carried out to prepare for a healthy and planned pregnancy and prevent HIV transmission in the family. Purpose: To explore phenomenologically the efforts to implement premarital screening in Jayapura Regency Method: The research approach is qualitative with a phenomenological research paradigm. Participants in this research were couples who were getting married, people in charge of the Infectious Disease Management program at the Community Health Center, and Persons in Charge of Infectious Disease Management at the Jayapura Regency Health Service. The sampling technique used a purposive sampling technique of 6 informants. Result: This research produced 3 themes, namely: premarital HIV screening in perception, the urgency of implementing premarital HIV screening and factors influencing the implementation of premarital screening. Perceptions of premarital screening were obtained from informants' statements regarding the understanding and purpose of the examination. From several categories, 3 themes were obtained, one of which was policy. Policies are obtained from 9 categories and 2 sub-themes schematically. Conclusion: The concept map resulting from the exploration of the premarital screening program in Jayapura Regency can be a reference or indicator in formulating a premarital screening program. Premarital screening program design can be carried out by analyzing the situation of need or urgency in Jayapura Regency. Suggestion: Support or strong reasons based on data are needed so that this program can be a recommendation for stakeholders.

Deep Learning-Based Blood Abnormalities Detection as a Tool for VEXAS Syndrome Screening.

VEXAS is a syndrome described in 2020, caused by mutations of the UBA1 gene, and displaying a large pleomorphic array of clinical and hematological features. Nevertheless, these criteria lack significance to discriminate VEXAS from other inflammatory conditions at the screening step. This work hence first focused on singling out dysplastic features indicative of the syndrome among peripheral blood (PB) polymorphonuclears (PMN). A deep learning algorithm is then proposed for automatic detection of these features. A multicentric dataset, comprising 9514 annotated PMN images was gathered, including UBA1 mutated VEXAS (n = 25), UBA1 wildtype myelodysplastic (n = 14), and UBA1 wildtype cytopenic patients (n = 25). Statistical analysis on a subset of patients was performed to screen for significant abnormalities. Detection of these features on PB was then automated with a convolutional neural network (CNN) for multilabel classification. Significant differences were observed in the proportions of PMNs with pseudo-Pelger, nuclear spikes, vacuoles, and hypogranularity between patients with VEXAS and both cytopenic and myelodysplastic controls. Automatic detection of these abnormalities yielded AUCs in the range [0.85-0.97] and a F1-score of 0.70 on the test set. A VEXAS screening score was proposed, leveraging the model outputs and predicting the UBA1 mutational status with 0.82 sensitivity and 0.71 specificity on the test patients. This study suggests that computer-assisted analysis of PB smears, focusing on suspected VEXAS cases, can provide valuable insights for determining which patients should undergo molecular testing. The presented deep learning approach can help hematologists direct their suspicions before initiating further analyses.

Rapid discovery and evolution of nanosensors containing fluorogenic amino acids

Binding-activated optical sensors are powerful tools for imaging, diagnostics, and biomolecular sensing. However, biosensor discovery is slow and requires tedious steps in rational design, screening, and characterization. Here we report on a platform that streamlines biosensor discovery and unlocks directed nanosensor evolution through genetically encodable fluorogenic amino acids (FgAAs). Building on the classical knowledge-based semisynthetic approach, we engineer ~15 kDa nanosensors that recognize specific proteins, peptides, and small molecules with up to 100-fold fluorescence increases and subsecond kinetics, allowing real-time and wash-free target sensing and live-cell bioimaging. An optimized genetic code expansion chemistry with FgAAs further enables rapid (~3 h) ribosomal nanosensor discovery via the cell-free translation of hundreds of candidates in parallel and directed nanosensor evolution with improved variant-specific sensitivities (up to ~250-fold) for SARS-CoV-2 antigens. Altogether, this platform could accelerate the discovery of fluorogenic nanosensors and pave the way to modify proteins with other non-standard functionalities for diverse applications.

Functional Aptamers In Vitro Evolution for Intranuclear Blockage of RNA-Protein Interaction.

Over the last 30 years, despite considerable research and endeavors aimed at harnessing aptamers as pharmaceutical molecules, the progress in developing aptamer-based drugs has been falling short of expectations. Sequential steps of affinity molecule acquisition and functional screening are typically required for discovering affinity-based macromolecule therapeutics, which can be time-consuming and limiting in candidate selection. Additionally, aptamers often necessitate tedious postselection modifications to overcome pharmacokinetic limitations, which usually impede the binding affinity. Herein, we propose a novel in vitro screening platform termed Functional Aptamers in vitro Evolution (FAIVE), which integrates affinity molecule acquisition with functional screening and introduces chemical diversity during the process. This platform aims to rapidly generate functional aptamers capable of binding to target proteins and regulating their functions. Illustrated by targeting intranuclear RNA-protein interactions involving HIV-1 Tat protein and TAR RNA, FAIVE demonstrates a selection of functional aptamers with significant intracellular blocking effects. The study also explores lipid nanoparticle delivery systems to enhance intracellular delivery efficiency, expanding aptamer targeting potential to broader intracellular and intranuclear domains. This study emphasizes the potential of FAIVE to expedite the development of aptamer-based drugs and facilitate the creation of more versatile and effective therapeutics.

Applications of cell free protein synthesis in protein design.

In protein design, the ultimate test of success is that the designs function as desired. Here, we discuss the utility of cell free protein synthesis (CFPS) as a rapid, convenient and versatile method to screen for activity. We champion the use of CFPS in screening potential designs. Compared to in vivo protein screening, a wider range of different activities can be evaluated using CFPS, and the scale on which it can easily be used-screening tens to hundreds of designed proteins-is ideally suited to current needs. Protein design using physics-based strategies tended to have a relatively low success rate, compared with current machine-learning based methods. Screening steps (such as yeast display) were often used to identify proteins that displayed the desired activity from many designs that were highly ranked computationally. We also describe how CFPS is well-suited to identify the reasons designs fail, which may include problems with transcription, translation, and solubility, in addition to not achieving the desired structure and function.

Towards Autonomous Living Meta-Analyses: A Framework for Automation of Systematic Review and Meta-Analyses.

Systematic review and meta-analysis constitute a staple of evidence-based medicine, an obligatory step in developing the guideline and recommendation document. It is a formalized process aiming at extracting and summarizing knowledge from the published work, grading, and considering the quality of the included studies. It is very laborious and time-consuming. Therefore, the meta-analyses are rarely updated and seldom living, decreasing their utility with time. Here, we present a framework for integrating the large language models and natural language processing techniques applied to the previously published systematic review and meta-analysis of the diagnostic test accuracy of the point of care tests. We show that the framework can be used to automate the screening step of the existing meta-analyses with minimal costs to quality and, to a large extent, the extraction step while maintaining the strict nature of the systematic review process.

Exploration of moisture activated dry granulation for the development of gastroretentive tablets aided by SeDeM diagram

Moisture activated dry granulation (MADG) is an attractive granulation process. However, only a few works have explored modified drug release achieved by MADG, and to the best of the authors knowledge, none of them have explored gastroretention. The aim of this study was to explore the applicability of MADG process for developing gastroretentive placebo tablets, aided by SeDeM diagram. Floating and swelling capacities have been identified as critical quality attributes (CQAs). After a formulation screening step, the type and concentration of floating matrix formers and of binders were identified as the most relevant critical material attributes (CMAs) to investigate in ten formulations. A multiple linear regression analysis (MLRA) was applied against the factors that were varied to find the design space. An optimized product based on principal component analysis (PCA) results and MLRA was prepared and characterized. The granulate was also assessed by SeDeM.In conclusion, granulates lead to floating tablets with short floating lag time (<2 min), long floating duration (>4 h), and showing good swelling characteristics. The results obtained so far are promising enough to consider MADG as an advantageous granulation method to obtain gastroretentive tablets or even other controlled delivery systems requiring a relatively high content of absorbent materials in their composition.

Ecotoxicological risk assessment on coagulation-flocculation in water/wastewater treatment: a systematic review.

It is undeniable that removal efficiency is the main factor in coagulation-flocculation (C-F) process for wastewater treatment. However, as far as environmental safety is concerned, the ecotoxicological aspect of the C-F process needs to be examined further. In this study, a systematic review was performed based on publications related to the toxicity research in C-F technology for wastewater treatment. Through a series of screening steps, available toxicity studies were categorized into four themes, namely acute toxicity, phytotoxicity, cytotoxicity, and genotoxicity, which comprised 48 articles. A compilation of the methodologies executed for each theme was also outlined. The findings show that conventional metallic coagulants (e.g., alum, iron chloride, and iron sulfate) were less toxic when tested on test species such as Daphnia magna (water flea), Lattuca sativa (lettuce), and animal cells compared to synthetic polymers. Natural coagulants such as chitosan or Moringa oleifera were less toxic compared to metallic coagulants; however, inconsistent results were observed. Moreover, an advanced C-F (electrocoagulation) as well as integration between C-F and Fenton, adsorption, and photocatalytic does not significantly change the toxicological profile of the system. It was found that diverse coagulants and flocculants, species sensitivity, complexity in toxicity testing, and dynamic environmental conditions were some key challenges faced in this field. Finally, it was expected that advances in technology, interdisciplinary collaboration, and a growing awareness of environmental sustainability will drive efforts to develop more effective and eco-friendly coagulants and flocculants, improve toxicity testing methodologies, and enhance the overall efficiency and safety of water and wastewater treatment processes.

Genomic Landscape of Branchio-Oto-Renal Syndrome through Whole-Genome Sequencing: A Single Rare Disease Center Experience in South Korea.

Branchio-oto-renal (BOR) and branchio-otic (BO) syndromes are characterized by anomalies affecting the ears, often accompanied by hearing loss, as well as abnormalities in the branchial arches and renal system. These syndromes exhibit a broad spectrum of phenotypes and a complex genomic landscape, with significant contributions from the EYA1 gene and the SIX gene family, including SIX1 and SIX5. Due to their diverse phenotypic presentations, which can overlap with other genetic syndromes, molecular genetic confirmation is essential. As sequencing technologies advance, whole-genome sequencing (WGS) is increasingly used in rare disease diagnostics. We explored the genomic landscape of 23 unrelated Korean families with typical or atypical BOR/BO syndrome using a stepwise approach: targeted panel sequencing and exome sequencing (Step 1), multiplex ligation-dependent probe amplification (MLPA) with copy number variation screening (Step 2), and WGS (Step 3). Integrating WGS into our diagnostic pipeline detected structure variations, including cryptic inversion and complex genomic rearrangement, eventually enhancing the diagnostic yield to 91%. Our findings expand the genomic architecture of BOR/BO syndrome and highlight the need for WGS to address the genetic diagnosis of clinically heterogeneous rare diseases.

Palliative care in patients with diabetes mellitus: integrative review

Objective: To synthesize scientific publications related to palliative care for patients with Diabetes Mellitus. Method: Integrative Review guided by the question “‘what is the influence of palliative care on the comfort, quality of life and glycemic control of patients with DM1 and DM2?”. Articles published in the last ten years (2013-2023), in 8 libraries and databases, were reviewed. Results: After searching and applying the inclusion and exclusion criteria, 6,443 were found and, after the screening and eligibility steps, 3 were included for analysis. Four thematic classes emerged: Reluctance and taboo in Palliative Care; Glycemic control and quality of life; Family and professional involvement; Reception and Health Care Networks in the context of Diabetes Mellitus. Conclusion: It is emphasized how beneficial early adherence to palliative care can be for a diabetic patient, aiming for better comfort and quality of life. Some factors limited the referral of these patients to palliative care: patient and family acceptance, resource limitations in palliative care programs and difficulties in pathological prognosis. Encouraging research related to this topic would significantly strengthen the debate on effective reception and better monitoring in palliative care.

Engineered reversible inhibition of SpyCatcher reactivity enables rapid generation of bispecific antibodies

The precise regulation of protein function is essential in biological systems and a key goal in chemical biology and protein engineering. Here, we describe a straightforward method to engineer functional control into the isopeptide bond-forming SpyTag/SpyCatcher protein ligation system. First, we perform a cysteine scan of the structured region of SpyCatcher. Except for two known reactive and catalytic residues, none of these mutations abolish reactivity. In a second screening step, we modify the cysteines with disulfide bond-forming small molecules. Here we identify 8 positions at which modifications strongly inhibit reactivity. This inhibition can be reversed by reducing agents. We call such a reversibly inhibitable SpyCatcher “SpyLock”. Using “BiLockCatcher”, a genetic fusion of wild-type SpyCatcher and SpyLock, and SpyTagged antibody fragments, we generate bispecific antibodies in a single, scalable format, facilitating the screening of a large number of antibody combinations. We demonstrate this approach by screening anti-PD-1/anti-PD-L1 bispecific antibodies using a cellular reporter assay.