LungLife AI (United States)

Abstract Background Flow cytometry is a cornerstone of immunological diagnostics, enabling precise assessment and monitoring of immunological diseases in clinical laboratories. The high throughput demands of modern diagnostics, however, often introduce challenges related to efficiency, reproducibility, and operator fatigue. The Curiox C-FREE™ Pluto System addresses these limitations by introducing a fully automated workflow, significantly reducing operator involvement. Methods Core features of the Curiox C-FREE™ Pluto System include non-disruptive sample handling, transferable protocols, and the elimination of aerosol generation and cross-contamination risks. These advancements enhance workflow reproducibility and operational safety. The integrated antibody cocktailing functionality further minimizes preparation errors and ensures staining consistency. Results In a representative clinical workflow of whole blood immunophenotyping, automation on Pluto eliminated 88 minutes of hands-on time per batch of 24 samples, resulting in a total workflow duration of 172 minutes for automation compared to 138 minutes for the conventional method. Comparative data analyses reveal that the system achieved population frequencies on par with traditional methods, while maintaining a robust 90% cell retention rate. Additionally, the system demonstrated superior debris removal, facilitating higher-quality data acquisition. Conclusion By combining efficiency, reproducibility, and safety, the Curiox C-FREE™ Pluto System represents a paradigm shift in clinical laboratory automation, supporting high-throughput diagnostics with precision and reliability.

Abstract Cancers, for example, non-small cell lung cancer (NSCLC) and colorectal cancer, are frequently associated with specific genetic mutations that serving as biomarkers to guide more specific treatments (targeted therapy). The most frequently mutated genes include EGFR, KRAS, NRAS, BRAF, HER2, PIK3CA, MET, etc. Unfortunately, screening and monitoring the most common cancer related mutations and identifying the breadth of the most common drug-resistant mutations in MRD monitoring has previously required NGS - but cost, and time for execution, limits testing frequency. Furthermore, sensitivity is typically 1-5% VAF, thus limiting early detection. While digital PCR (dPCR) can be run at a fraction of the cost, and with higher sensitivity (VAFs of 0.1 to 0.01%), due to the lack of high level of multiplexing, it was historically impractical for routine screening. Here, we show that the use 6-color naica® system (Stilla Technologies), incorporating a series of high multiplex panels developed by Atila BioSystems, is able to group and efficiently detect the most frequently druggable mutations in a single reaction within three hours. Furthermore, we are demonstrating the coverage, precision, and sensitivity necessary to make high-plex digital PCR a viable option for future liquid biopsy testing. Citation Format: Michael Granitsiotis, Kimberly Gutierrez, Youxiang Wang, Jialuo Li, Zhijie Yang. Enabling higher sensitivity liquid biopsies with multiplex digital PCR NSCLC-relevant drug resistance panels [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1048.

BackgroundThere is currently a need for additional diagnostic information to help guide treatment decisions and to properly determine the best treatment pathway for patients identified with indeterminate pulmonary nodules (IPNs). The aim of this study was to demonstrate the incremental cost-effectiveness of LungLB compared to the current clinical diagnostic pathway (CDP) in the management of patients with IPNs, from a US payer’s perspective.MethodsA decision tree and Markov model hybrid was chosen from a payer perspective in the US setting, based on published literature, to assess the incremental cost-effectiveness of LungLB compared to the current CDP in the management of patients with IPNs. Primary endpoints of the analysis include expected costs, life years (LYs), and quality-adjusted life years (QALYs) for each arm of the model, as well as an incremental cost-effectiveness ratio (ICER), which is calculated as the incremental costs per QALY, and net monetary benefit (NMB).ResultsWe find that, with the inclusion of LungLB to the current CDP diagnostic pathway, expected LYs over the typical patient’s lifespan increase by 0.07 years and QALYs increase by 0.06. The average patient in the CDP arm will pay approximately $44,310 over their lifespan, while a patient in the LungLB arm will pay $48,492, resulting in a difference of $4,182. The differentials between the CDP and LungLB arms of the model in costs and QALYs yield an ICER of $75,740 per QALY and an incremental NMB of $1,339.ConclusionThis analysis provides evidence that LungLB, in conjunction with CDP, is a cost-effective alternative compared to the current CDP alone in a US setting for individuals with IPNs.

Topical drug administration in the treatment of skin disorders is known to have high efficacy with a favorable safety profile in clinical settings. Disposition of the active pharmaceutical ingredient (API) in the skin layers plays a critical role for the dose regimen calculator required for desired therapy efficiency. Experimental measurements of the API skin concentration are complicated and inaccurate and can be replaced with physiologically based pharmacokinetic modeling predicting local and systemic drug exposure. Opioid analgesics are commonly used in treating severe pain and it is frequently associated with putative adverse effects. Topical applications of opioids were shown to have high efficacy with a favorable safety profile in clinical settings. Clinical investigations of transdermal drug formulations, especially in patients with skin lesions, are often hindered due to the patient intolerance, high cost of the studies, and ethical aspects.BIOiSIM, a hybrid artificial intelligence computational platform integrating machine learning optimization with mechanistic modeling was implemented to accurately simulate in vivo plasma concentration profile and predict partition coefficients and API skin retention for two opioid analgesics in healthy skin and simulated skin lesions. Publicly available drug parameters and physiological data were used for the model building.The platform successfully predicted API systemic exposure and skin concentration following the epicutaneous application of central opioid agonist buprenorphine and peripheral opioid agonist oxycodone in healthy human skin and simulated subjects impaired skin. Experimentally observed plasma concentration for both drugs demonstrated validity of the in‐silico methods. The results indicated that buprenorphine permeates rapidly through the stratum corneum and partitions into the dermis, where its concentration slowly declines as the reservoir in the compound vehicle is depleted. Thus, the impact of lesions is not as significant. In contrast, the hydrophilic oxycodone, where the rate‐limiting stratum corneum layer prevents a significant portion of the compound from permeating deeper in healthy patients. Hence, concentration of oxycodone sharply declines over the first hours of exposure in deeper indicating poorer accumulation capacity in the skin in the presence of lesions.An AI/ML‐driven PBPK model was applied for accurate simulation skin concentrations of two opioid analgesics – buprenorphine and oxycodone. The results have confirmed that both these drugs have good skin retention and can stimulate opioid receptors located in skin. The results suggest that buprenorphine showing substantial skin retention capacity can be applied topically for the treatment of skin disorders in in patients.

Glucose transporter type 1 deficiency syndrome (Glut1DS) is a treatable genetic neurometabolic disorder because of a variant in the Glut1 transporter, encoded by the SLC2A1 gene (OMIM*138140), resulting in impaired glucose transport across the blood–brain barrier. Glut1DS was initially reported in infants with progressive encephalopathy, acquired microcephaly, and drug-resistant epilepsy and can be treated with the ketogenic diet.1 With increased awareness and progress in diagnosis, the phenotype has expanded to include milder presentations of epilepsy, movement disorders, and cognitive impairment.2 Glut1DS is estimated to affect 1 in every 24,000 live births.3 A 5-year-old Spanish girl presented with paroxysmal movement disorder, characterized by abnormal leg posture, unsteady gait, and frequent falls by 2 years and 8 months of age. Five months later, she had an episode that lasted 4 hours. Subsequently, frequent episodes were reported to occur on a weekly basis. One of those episodes included dysarthria without loss of consciousness (Video 1). Given her age, her family has been unable to identify a specific trigger for these episodes. She was born to term after a normal pregnancy to healthy non-consanguineous Caucasian parents with a positive family history of epilepsy (maternal grandmother). Her early neurodevelopment was described as normal, with a slight delay in independent gait acquisition at 18 months. A neurological examination at 5 years and 7 months of age showed microcephaly (cranial circumference of 48 cm, 2nd percentile); although no other neurological signs were observed. During hospitalization, a brain MRI was performed with normal results. Video electroencephalogram (EEG) showed electroclinical myoclonic seizures. These epileptic myoclonus occurred infrequently and were not identified by the family as seizures. She had no other types of seizures. Extensive biochemical and metabolic tests showed normal results. Lumbar puncture showed hypoglycorrhachia of 32 mg/dL and a cerebrospinal fluid (CSF)/serum glucose ratio of 0.5 (reference value >0.60) and flow cytometric analysis of the surface expression of Glut1 in circulating erythrocytes was reduced to 36% compared to the general population according to the METAglut1 test (Fig. 1). Both tests were helpful in confirming the diagnostic suspicion of Glut1DS. Following that, whole exome sequencing revealed a previously reported heterozygous de novo variant on the SLC2A1 gene, p.Arg333Gln/c.998G>A. A ketogenic diet with a fat-to-protein plus carbohydrate ratio of 4:1 was started with good tolerance. In clinical follow-up after 24 months of ketogenic diet treatment, no further episodes of paroxysmal movement disorder or myoclonic seizures were reported. The video EEG did not report any epileptic abnormalities or seizures. Her neuropsychological evaluation is described in Appendix S1. The legal guardians gave their consent for publication and the study was approved by the Ethics Committee (ART-07-21). Glut1DS is a well-known clinical entity comprising a classical (90%) and a non-classical (10%) phenotype. Non-epileptic paroxysmal events, such as those presented by our patient, have been detected in 28% of a series of 57 patients with Glut1DS.4 Although paroxysmal exercise-induced dyskinesia is the most common type of paroxysmal dyskinesia in Glut1DS, other non-kinesigenic dyskinesia, episodic ataxia, or a combination of subtypes can also be present (Video 1).5 The diagnosis of Glut1DS is established with a CSF glucose concentration of <60 mg/dL in combination with a normal blood glucose concentration and with the identification of a heterozygous pathogenic variant in the SLC2A1 gene by molecular genetic testing. However, the decrease of more than 20% in Glut1 expression on the surface of the erythrocytes detected by the METAglut1 test compared to the general population is also highly suggestive.6 Gras et al,6 first introduced the METAglut1 as a simple diagnostic blood test for Glut1DS using flow cytometry, with a detection rate of 78% in patients with proven Glut1DS. An EDTA tube is used to obtain whole blood for the METAglut1 test. The patient does not need to fast before the blood draw. The sample is sent to the central lab at 4°C and results are available in 48 hours. The assay takes approximately 1 hour to run and analyze 1 to 18 samples. Glut1 expression is measured directly on erythrocytes using a simple flow cytometer.6 Currently, the test is only available in France but it is expected to be implemented soon in a number of European countries with the CE mark (Spain, Italy, and Germany). The fact that METAglut1 can be performed using blood analysis rather than a lumbar puncture and that it takes only a short time to perform compared to months for a genetic study in a hospital setting, clearly demonstrates its use in screening for Glut1DS. The latest international Glut1DS study group review concluded that the METAglut1 test is not yet a standard diagnostic tool for Glut1DS,7 whereas ongoing studies should provide actionable results soon (NCT 03722212). In conclusion, our report describes a case of non-kinesigenic paroxysmal dyskinesias associated with Glut1DS and highlights METAglut1 test use for rapid disease diagnosis. The authors thank the patient and her family. (1) Research project: A. Conception, B. Organization, C. Execution; (2) Statistical Analysis: A. Design, B. Execution, C. Review and Critique; (3) Manuscript: A. Writing of the First Draft, B. Review and Critique. L.S.: 3A, 3B L.M.: 1C, 3B D.Y.: 1C, 3B C.V.: 1C, 3B V.P.: 1C, 2A, 3B J.D.O.E.: 1A, 1B, 1C, 3B The legal guardians gave their written consent to the recording of the patient for publication, and the study received ethical approval by the Ethics Committee (ART-07-21). We confirm that we have read the Journal's position on issues involved in ethical publication and affirm that this work is consistent with those guidelines. No specific funding was received for this work. V.P. is the cofounder and CEO of Metafora Biosystems. The authors declare that there are no other conflicts of interest relevant to this work. The authors declare that there are no additional disclosures to report. Appendix S1. Neuropsychological evaluation. (A) Kaufman K-BIT Brief Intelligence Test. The global composite IQ score is 94, placing the intellectual capabilities of the patient at a medium level rather than the expected level for her age. (B) Vineland-II Interview. The patient achieves appropriate sociability scores, but deviates from normal communication, daily skills, and psychomotor development. (C) Navarra oral language test (PLON). The PLON results suggest that the linguistic content and use are within normal limits. The findings of the language form section demonstrate that certain features of morphology and syntax do not develop with age. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Purpose: The neonatal period represents a unique stage of life when critical foundations of lifelong health are established. Throughout this period, the gut microbiome provides protection against infections and is crucial for the proper development of the immune system. Deviations from optimal microbiome compositions during the postnatal stage, conceptually referred to as dysbiosis, are associated with the absence of key infant-associated Bifidobacterium known to facilitate the utilization of human milk oligosaccharides (HMOs), increased enteric inflammation, and an increase in potentially pathogenic taxa. Although previous studies have revealed a marked dysbiosis …

EndoBI-1 and EndoBI-2 are two endo- β-N- acetylglucosaminidase isoenzymes that cleave N-N’- diacetylchitobiosyl moieties found in various types of native N -glycans. These N -glycans are indigestible by human infants and adults due to the lack of responsible glycosyl hydrolases and they act as selective prebiotics for a probiotic microorganism, Bifidobacterium longum subsp . infantis , in the large intestine. The selectivity and the thermostability of EndoBI-1 and EndoBI-2 suggest that these enzymes may be useful for many scientific and industrial applications. In this study, the growing numbers of homologous sequences in different databases were exploited in a comparative approach to investigate structural properties of EndoBI-1 and EndoBI-2 enzymes. Moreover, the complete and partial homology models of these two enzymes were generated and evaluated. Selected models were used for docking studies of the plus subsite ligand of these enzymes for further understanding on the substrate selectivity of EndoBI enzymes.

SUMMARYImmune-microbe interactions early in life influence an individual’s risk of developing allergies, asthma and some autoimmune disorders. Breastfeeding helps guide the development of healthy immune-microbe relationships, in part by providing nutrients to specialized microbes that in turn benefit the host and its developing immune system. Such bacteria having co-evolved with humans are associated with reduced risks of immune mediated diseases but are increasingly rare in modern societies. Here we map an immunological sequence of events, triggered by microbial colonization that distinguish children with different gut bacterial composition. Lack of bifidobacterial species is associated with elevated markers of intestinal inflammation and immune dysregulation and in a randomized trial of breastfed infants, the infant-adaptedBifidobacterium infantisEVC001 silenced intestinal Th2 and Th17 immune responses, while inducing IFNβ, and its metabolites skew T-cell polarizationin vitro, from Th2 towards Th1, suggesting a healthier immune imprinting during the first critical months of life.HIGHLIGHTSAn ordered sequence of immune changes after birth, driven by microbial interactionsLow gutBifidobacteriumabundance is associated with markers of intestinal inflammationFeedingB. infantisEVC001 silenced intestinal Th2 and Th17 but upregulates IFNβB. infantisEVC001 metabolites and/or enteric cytokines skew naïve T-cell polarization towards Th1 and away from Th2

Cross-feeding based on the metabolite 1,2-propanediol has been proposed to have an important role in the establishment of trophic interactions among gut symbionts, but its ecological importance has not been empirically established. Here, we show that in vitro growth of Lactobacillus reuteri (syn. Limosilactobacillus reuteri) ATCC PTA 6475 is enhanced through 1,2-propanediol produced by Bifidobacterium breve UCC2003 and Escherichia coli MG1655 from the metabolization of fucose and rhamnose, respectively. Work with isogenic mutants showed that the trophic interaction is dependent on the pduCDE operon in L. reuteri, which encodes the ability to use 1,2-propanediol, and the l-fucose permease (fucP) gene in B. breve, which is required for 1,2-propanediol formation from fucose. Experiments in gnotobiotic mice revealed that, although the pduCDE operon bestows a fitness burden on L. reuteri ATCC PTA 6475 in the mouse digestive tract, the ecological performance of the strain was enhanced in the presence of B. breve UCC2003 and the mucus-degrading species Bifidobacterium bifidum The use of the respective pduCDE and fucP mutants of L. reuteri and B. breve in the mouse experiments indicated that the trophic interaction was specifically based on 1,2-propanediol. Overall, our work established the ecological importance of cross-feeding relationships based on 1,2-propanediol for the fitness of a bacterial symbiont in the vertebrate gut.IMPORTANCE Through experiments in gnotobiotic mice that employed isogenic mutants of bacterial strains that produce (Bifidobacterium breve) and utilize (Lactobacillus reuteri) 1,2-propanediol, this study provides mechanistic insight into the ecological ramifications of a trophic interaction between gut symbionts. The findings improve our understanding on how cross-feeding influences the competitive fitness of L. reuteri in the vertebrate gut and revealed a putative selective force that shaped the evolution of the species. The findings are relevant since they provide a basis to design rational microbial-based strategies to modulate gut ecosystems, which could employ mixtures of bacterial strains that establish trophic interactions or a personalized approach based on the ability of a resident microbiota to provide resources for the incoming microbe.