Keeping an "eye" on the experiment: computer vision for real-time monitoring and control.

This work presents a generalizable computer vision (CV) and machine learning model that is used for automated real-time monitoring and control of a diverse array of workup processes. Our system simultaneously monitors multiple physical outputs (e.g., liquid level, homogeneity, turbidity, solid, residue, and color), offering a method for rapid data acquisition and deeper analysis from multiple visual cues. We demonstrate a single platform (consisting of CV, machine learning, real-time monitoring techniques, and flexible hardware) to monitor and control vision-based experimental techniques, including solvent exchange distillation, antisolvent crystallization, evaporative crystallization, cooling crystallization, solid-liquid mixing, and liquid-liquid extraction. Both qualitative (video capturing) and quantitative data (visual outputs measurement) were obtained which provided a method for data cross-validation. Our CV model's ease of use, generalizability, and non-invasiveness make it an appealing complementary option to in situ and real-time analytical monitoring tools and mathematical modeling. Additionally, our platform is integrated with Mettler-Toledo's iControl software, which acts as a centralized system for real-time data collection, visualization, and storage. With consistent data representation and infrastructure, we were able to efficiently transfer the technology and reproduce results between different labs. This ability to easily monitor and respond to the dynamic situational changes of the experiments is pivotal to enabling future flexible automation workflows.

Therapeutic Antibodies in Medicine.

Antibody engineering has developed into a wide-reaching field, impacting a multitude of industries, most notably healthcare and diagnostics. The seminal work on developing the first monoclonal antibody four decades ago has witnessed exponential growth in the last 10-15 years, where regulators have approved monoclonal antibodies as therapeutics and for several diagnostic applications, including the remarkable attention it garnered during the pandemic. In recent years, antibodies have become the fastest-growing class of biological drugs approved for the treatment of a wide range of diseases, from cancer to autoimmune conditions. This review discusses the field of therapeutic antibodies as it stands today. It summarizes and outlines the clinical relevance and application of therapeutic antibodies in treating a landscape of diseases in different disciplines of medicine. It discusses the nomenclature, various approaches to antibody therapies, and the evolution of antibody therapeutics. It also discusses the risk profile and adverse immune reactions associated with the antibodies and sheds light on future applications and perspectives in antibody drug discovery.

CRISPR-Directed Therapeutic Correction at the NCF1 Locus Is Challenged by Frequent Incidence of Chromosomal Deletions.

Resurrection of non-processed pseudogenes may increase the efficacy of therapeutic gene editing, upon simultaneous targeting of a mutated gene and its highly homologous pseudogenes. To investigate the potency of this approach for clinical gene therapy of human diseases, we corrected a pseudogene-associated disorder, the immunodeficiency p47 phox -deficient chronic granulomatous disease (p47 phox CGD), using clustered regularly interspaced short palindromic repeats-associated nuclease Cas9 (CRISPR-Cas9) to target mutated neutrophil cytosolic factor 1 (NCF1). Being separated by less than two million base pairs, NCF1 and two pseudogenes are closely co-localized on chromosome 7. In healthy people, a two-nucleotide GT deletion (ΔGT) is present in the NCF1B and NCF1C pseudogenes only. In the majority of patients with p47 phox CGD, the NCF1 gene is inactivated due to a ΔGT transfer from one of the two non-processed pseudogenes. Here we demonstrate that concurrent targeting and correction of mutated NCF1 and its pseudogenes results in therapeutic CGD phenotype correction, but also causes potentially harmful chromosomal deletions between the targeted loci in a p47 phox -deficient CGD cell line model. Therefore, development of genome-editing-based treatment of pseudogene-related disorders mandates thorough safety examination, as well as technological advances, limiting concurrent induction of multiple double-strand breaks on a single chromosome.

Systemic administration of an alpha-7 nicotinic acetylcholine agonist reverses neuropathic pain in male Sprague Dawley rats.

UNLABELLED: Alpha-7 nicotinic acetylcholine receptor (α7 nAChR) agonists attenuate pain and inflammation in preclinical models. This study tested whether systemic delivery of an α7 nAChR agonist attenuates neuropathic pain and associated immune-mediated pro-inflammation. Hind paw response thresholds to mechanical stimuli in male Sprague Dawley rats were assessed before and after sciatic chronic constriction injury (CCI) or sham surgery. Osmotic mini-pumps containing TC-7020, an α7 nAChR selective agonist, were implanted 10 to 14 days after surgery. TC-7020 (1, 3, and 10 mg/kg/d; s.c.) significantly attenuated CCI-induced allodynia, which lasted through 2 weeks of test compound administration. Spinal cords were collected after 2 weeks and processed for microglial and astrocyte activation markers within the ipsilateral L4-L6 dorsal horn. In addition, ipsilateral L4-5 dorsal root ganglia (DRGs) were processed for neuronal injury and satellite cell activation markers. CCI-induced central glial cell activation markers were not suppressed by TC-7020, even though TC-7020 is mildly blood-brain barrier permeable. However, TC-7020 downregulated the integrated density of activation transcription factor 3 (ATF3) but not the number of ATF positive cells. TC-7020 also downregulated phosphorylated extracellular signal kinase (p-ERK) and satellite cell activation in the CCI-affected DRGs. Therefore, systemic α7 nAChR agonist may be effective in treating neuropathic pain via reducing neuronal injury and immune cells activation occurring in the periphery. PERSPECTIVE: These studies demonstrated that TC-7020, an alpha7 nicotinic acetylcholine receptor agonist with partial blood-brain barrier permeability, reversed neuropathic pain in rats, likely via attenuation of inflammation in the DRG and/or the site of sciatic injury.

The productivity measurement and enhancement system: a meta-analysis.

Meta-analytic procedures were used to examine data from 83 field studies of the Productivity Measurement and Enhancement System (ProMES). The article expands the evidence on effectiveness of the intervention, examines where it has been successful, and explores moderators related to its success. Four research questions were explored and results indicate that (a) ProMES results in large improvements in productivity; (b) these effects last over time, in some cases years; (c) the intervention results in productivity improvements in many different types of settings (i.e., type of organization, type of work, type of worker, country); and (d) moderator variables are related to the degree of productivity improvement. These moderator variables include how closely the study followed the original ProMES methodology, the quality of feedback given, whether changes were made in the feedback system, the degree of interdependence of the work group, and centralization of the organization. Implications based on these findings are discussed for future use of this intervention, and the system is discussed as an example for evidence-based management.

Phaeohyphomycotic soft tissue infections caused by the coelomycetous fungus Microsphaeropsis arundinis.

Microsphaeropsis arundinis is an anamorphic fungal plant inhabitant belonging to the form class Coelomycetes. We describe two cases of M. arundinis soft tissue infections in immunosuppressed patients. This organism has not previously been described as causing disease in humans. It was identified on the basis of its typical ostiolate pycnidial conidiomata, ampulliform conidiogenous cells, and small, smooth-walled, brown, cylindrical conidia.

Isolation and identification of Burkholderia cepacia by participants in an external Quality Assurance Program (QAP) between 1994 and 1999.

AIM: External quality assurance programs (QAPs) provide an opportunity to benchmark laboratory performance according to the profile of specimens received. Participant confidentiality is maintained within each group of laboratories whose performance is measured using similar, repetitive exercises. Isolation and identification of Burkholderia cepacia from simulated cystic fibrosis (CF) sputa was a clinically relevant exercise that provided a model for this analytical approach. METHODS: Between 1994 and 1999, six Royal College of Pathologists of Australasia (RCPA) Microbiology QAPs included four simulated CF sputa and two panels of oxidative Gram-negative bacilli. Laboratories were grouped according to experience with CF sputa disclosed by two questionnaires. Data were analysed by laboratory group for ability to isolate and identify B. cepacia. RESULTS: Three laboratory groups annually received >100 CF sputa (CF>100), 100 CF sputa or fewer, or did not regularly receive CF sputa. CF>100 laboratories inoculated more isolation media, were more likely to use selective media and were less likely to misidentify B. cepacia than the other groups. Improved performance by CF>100 laboratories was marked after the first exercise and remained at a high level compared with the other two groups. This trend in performance was also apparent for Pseudomonas aeruginosa although the numbers of errors were less than for B. cepacia. CONCLUSIONS: These exercises demonstrated consistently improved performance only among CF>100 laboratories. The future criteria for laboratory accreditation may include performance as well as participation in QAPs, placing additional burdens on organisers and participants.