Compatible Protocol Research Articles

Comparative analysis of iPSC-derived NK cells from two differentiation strategies reveals distinct signatures and cytotoxic activities.

The ability to generate natural killer (NK) cells from induced pluripotent stem cells (iPSCs) has given rise to new possibilities for the large-scale production of homogeneous immunotherapeutic cellular products and opened new avenues towards the creation of "off-the-shelf" cancer immunotherapies. However, the differentiation of NK cells from iPSCs remains poorly understood, particularly regarding the ontogenic landscape of iPSC-derived NK (iNK) cells produced in vitro and the influence that the differentiation strategy employed may have on the iNK profile. To investigate this question, we conducted a comparative analysis of two sets of iNK cells generated from the same iPSC line using two different protocols: (i) a short-term, clinically compatible feeder-free protocol corresponding to primitive hematopoiesis, and (ii) a lymphoid-based protocol representing the definitive hematopoietic step. Our work demonstrated that both protocols are capable of producing functional iNK cells. However, the two sets of resulting iNKs exhibited distinct phenotypes and transcriptomic profiles. The lymphoid-based differentiation approach generated iNKs with a more mature and activated profile, which demonstrated higher cytotoxicity against cancer cell lines compared to iNK cells produced under short-term feeder-free conditions suggesting that the differentiation strategy must be considered when designing iNK cell-based adoptive immunotherapies.

Architecting Compatible PIM Protocol for CPU-PIM Collaboration

Architecting Compatible PIM Protocol for CPU-PIM Collaboration

Use of neurological criteria to declare death in children.

Accurate determination of death is a necessary responsibility of the medical profession. Brain death, or death by neurological criteria (DNC), can be legally declared after the determination of permanent loss of clinical brain function, including the capacity for consciousness, brainstem reflexes, and the ability to breathe spontaneously. Despite longstanding debates over the exact definition of brain death or DNC and how it is determined, most middle- and high-income countries have compatible medical protocols and legal policies for brain death or DNC. This review summarizes the 2023 updated guidelines for brain death or DNC determination, which integrate adult and pediatric diagnostic criteria. We discuss the clinical challenges related to brain death or DNC determination in infants and young children. We emphasize that physicians must follow the standardized and meticulous evaluation processes outlined in these guidelines to reduce diagnostic error and ensure no false positive determinations. An essential component of the brain death or DNC evaluation is appropriate and transparent communication with families. Ongoing efforts to promote consistency and legal uniformity in the declaration of death are needed.

Chemoenzymatic Dynamic Kinetic Resolution Protocol with an Immobilized Oxovanadium as a Racemization Catalyst.

An excellent compatible and cost-effective dynamic kinetic resolution (DKR) protocol has been developed by combining a novel immobilized oxovanadium racemization catalyst onto cheap diatomite (V-D) with an immobilized lipase LA resolution catalyst onto a macroporous resin (LA-MR). V-D was prepared via grinding immobilization, which may become a promising alternative for the immobilization of metals, especially precious metals due to its low cost, high efficiency, easy separation, and large reaction interface. The DKR afforded high yield (96.1%), e.e. (98.67%), and Sel (98.28%) under optimal conditions established using response surface methodology as follows: the amount of V-D 10.83 mg, reaction time 51.2 h, and temperature 48.1 °C, respectively, indicating that all the reactions in the DKR were coordinated very well. The DKR protocol was also found to have high stability up to six reuses. V-D exhibited excellent compatibility with LA-MR because the lipase immobilized onto MR did not physically contact with the vanadium species immobilized onto diatomite, thus avoiding inactivation. Considering that lipase, oxovanadium, diatomite, and MR used are relatively inexpensive, and the adsorption or grinding immobilization is simple, the LA-V-MD DKR by coupling LA-MR with V-D is a cost-effective and promising protocol for chiral secondary alcohols.

RETENTIVE STRENGTH OF CEMENTRETAINED IMPLANT-SUPPORTED FIXED DENTAL PROSTHESES ACCORDING TO DIFFERENT CEMENT TYPES AND CEMENTATION PROTOCOLS: AN IN VITRO STUDY.

To develop the most compatible cementation protocol for ensuring minimal residual cement and optimal retention of cement-retained implant-supported fixed dental prostheses. A total of 30 custom implant abutments and zirconia crowns with bilateral wings were prepared. Three cement types were used for cementation: noneugenol resin cement (Premier Implant Cement; Group IC), dual-polymerizing self-adhesive resin cement (SmartCem2; Group SC), and zinc oxide eugenol cement (Temp-Bond; Group TB; n = 30 per group). Three cementation methods were applied for each cement type, and the samples were divided into subgroups: (1) cement was injected using a graduated syringe (ICN, SC-N, and TB-N); (2) a cementa/on jig made with a silicone impression material and temporary resin material was used (IC-CJ, SC-CJ, and TB-CJ); (3) 3D-printed abutments were used as replicas for cementation (IC-3DP, SC-3DP, and TB-3DP). The amount of cement injected, surface area of the residual cement, and retentive strength were measured. Kruskal- Wallis and post-hoc Mann-Whitney tests were used for statistical analyses. Excess cement was not observed when cementa/on jig or 3D-printed replicas were used. For IC and SC subgroups, non-use of these auxiliary tools resulted in significantly higher amounts of injected cement. The retentive strength differed significantly among the IC subgroups but not among the SC subgroups. The retentive strength of subgroups TB-N and TB-CJ was significantly higher than that of subgroup TB-3DP. To prolong the main purpose of each cement type, a cementation jig or 3D-printed replica is highly recommended regardless of the cement type.

Recipient microbiome-related features predicting metabolic improvement following fecal microbiota transplantation in adults with severe obesity and metabolic syndrome: a secondary analysis of a phase 2 clinical trial

ABSTRACT Microbial-based therapeutics in clinical practice are of considerable interest, and a recent study demonstrated fecal microbial transplantation (FMT) followed by dietary fiber supplements improved glucose homeostasis. Previous evidence suggests that donor and recipient compatibility and FMT protocol are key determinants, but little is known about the involvement of specific recipient factors. Using data from our recent randomized placebo-control phase 2 clinical trial in adults with obesity and metabolic syndrome, we grouped participants that received FMT from one of 4 donors with either fiber supplement into HOMA-IR responders (n = 21) and HOMA-IR non-responders (n = 8). We further assessed plasma bile acids using targeted metabolomics and performed subgroup analyzes to evaluate the effects of recipient parameters and gastrointestinal factors on microbiota engraftment and homeostatic model assessment of insulin resistance (HOMA2-IR) response. The baseline fecal microbiota composition at genus level of recipients could predict the improvements in HOMA2-IR at week 6 (ROC-AUC = 0.70). Prevotella was identified as an important predictor, with responders having significantly lower relative abundance than non-responders (p = .02). In addition, recipients displayed a highly individualized degree of microbial engraftment from donors. Compared to the non-responders, the responders had significantly increased bacterial richness (Chao1) after FMT and a more consistent engraftment of donor-specific bacteria ASVs (amplicon sequence variants) such as Faecalibacillus intestinalis (ASV44), Roseburia spp. (ASV103), and Christensenellaceae spp. (ASV140) (p < .05). Microbiota engraftment was strongly associated with recipients’ factors at baseline including initial gut microbial diversity, fiber and nutrient intakes, inflammatory markers, and bile acid derivative levels. This study identified that responders to FMT therapy had a higher engraftment rate in the transplantation of specific donor-specific microbes, which were strongly correlated with insulin sensitivity improvements. Further, the recipient baseline gut microbiota and related factors were identified as the determinants for responsiveness to FMT and fiber supplementation. The findings provide a basis for the development of precision microbial therapeutics for the treatment of metabolic syndrome.

Towards developing a matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) compatible tissue expansion protocol

Mass spectrometry imaging (MSI) visualizes spatial distribution of molecules in a biological tissue. However, compared with traditional microscopy-based imaging, conventional MSI is limited to its spatial resolution, resulting in difficulties in identifying detailed tissue morphological characters, such as lesion boundary or nanoscale structures. On the other hand, expansion microscopy, a tissue expansion method widely used in optical imaging to improve morphological details, has great potential to solve insufficient spatial resolution in mass spectrometry imaging (MSI). However, expansion microscopy was not originally designed for MSI, resulting in problems while combining expansion microscopy and MSI such as expanded sample fragility, vacuum stability and molecule loss during sample preparation. In this research we developed a MALDI MSI compatible expansion protocol by adjusting sample preparation methods during tissue expansion, successfully combining expansion microscopy with MSI. After tissue expansion the expanded sample can be readily applied to MALDI MSI sample preparation and further data acquisition. The MALDI MSI compatible expansion protocol has great potential to be widely applied in MALDI MSI sample preparation to facilitate improvement of MSI spatial resolution.

Indocyanine Green-Labeled Platelets for Survival and Recovery Studies

Plain Language SummaryWe developed a simple, reproducible method according to Good Manufacturing Practice guidelines for labeling of platelets from platelet concentrates (PCs) with indocyanine green (ICG) as a non-radioactive alternative. PCs are medicinal drugs that are transfused to prevent or treat bleeding. They consist of the blood cells’ platelets which are responsible for clotting processes in the body. Manufacturing procedures of PCs are continuously refined, and for in vivo testing, these platelets have to be labeled to track and to distinguish them from proband’s or patient’s own cells. Radioactive labeling, for a long time the gold standard for cell labeling, is no longer accepted. ICG is a fluorescent dye approved by the drug authorities and already used for diagnostic purposes in humans. We used ICG to label platelets from PCs. With our method, we achieved a labeling efficiency of 99.8%. We used whole blood (WB) as an ex vivo matrix to monitor the labeling stability of ICG-labeled platelets. After the addition of ICG-labeled platelets to WB, the labeling efficiency decreased to 81% after 24 h. However, we were still able to distinguish the ICG-labeled platelets from the WB platelets. We could also show that platelet function was not impaired by the labeling processes. The good tolerance of ICG indicates a short path to clinical application in healthy volunteers and investigations of novel PC-manufacturing procedures. As a read-out system, flow cytometry systems equipped with NIR lasers and filters could offer the possibility of rapid visualization, cell tracking, re-isolation, and ex vivo studies.

Synthesis, Structural, Spectral Studies and Activity Studies of 6-Methyl-4-(3-Nitro-Phenyl)-2-Oxo-1,2,3,4 Tetrahydropyrimidine-5-Carboxylic Acid Ethyl Ester Catalyzed Using BF3-OEt2

Amide compounds exhibit tautomerism which is an important property in organic synthesis. Evidence for the occurrence of tautomeric forms for amido-imidols is interpreted via mass spectrometry techniques. The tautomeric equilibrium of pyrimidone moiety was rationalized and the existence of amido-imidol tautomerism in 6-methyl-4-(3-nitro-phenyl)-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid ethyl ester (6M3NP) was verified. The proton nuclear magnetic resonance splitting pattern shows the intramolecular hydrogen bonding between the keto-enol forms which evidences through a fluctuation in the integration value of C4 proton. Synthesis proceeds through novel BF3-OEt2 catalysed one-pot, simple, efficient, minimal catalyst usage, compatible, outstanding yield, and rapid protocol for the Tetrahydropyrimidinones via Biginelli reaction is described. The titled compound 6M3NP was characterized by infrared and nuclear magnetic resonance. The compound was screened for antimicrobial activity against 3 microbes which showed a high zone of inhibition when compared to standard streptomycin. The antioxidant activity using 2,2-diphenyl-1-picryl-hydrazyl-hydrate assay gave a promising IC50 value of 14.11 µM. The more the radical scavenging activity the greater the chance of functioning as an anticancer agent. 6M3NP was tested against breast cancer cells after scrutiny with 60 cell lines. Clonogenicity and soft agar colony formation were significantly reduced in MCF7, MDA-MB-231, and T47D breast cancer cells after treatment with 6M3NP. The in-silico dock score obtained for the title compound was 62.845 which is 40% more than the standard drug Nifedipine. In-vitro cell line analysis results showed that the title compound possesses maximum activity than commercial drug Nifedipine against cardiovascular disease as a calcium channel blocker.

Sistem Komunikasi Mikrokontroler dan PLC Berbasis Komunikasi Serial Host Link dan Protokol C-Command RS232

The communication between microcontrollers and PLC has become an important technology development that needs to be enhanced. One of the reasons for the communication between these two devices is to facilitate the input process of data or information from sensors, which are responsible for detecting or measuring specific parameters. Not all sensors can be directly read by PLC, as some sensors have their own communication protocols or specific libraries that are not directly supported by PLC. Microcontrollers are often used as intermediaries between sensors and PLC. Microcontrollers can read sensor data and then convert it into a format that can be understood by PLC. Microcontrollers can perform flat processing tasks such as conversion and status evaluation to provide more relevant data, and then send this data to the PLC through a compatible communication protocol. In this study, the communication protocol used is the Host Link serial communication using the C-mode Command method. The communication testing conducted using 10 different data samples resulted in accurate values with an error percentage of 0%.

Characterizing Intraindividual Podocyte Morphology In Vitro with Different Innovative Microscopic and Spectroscopic Techniques.

Podocytes are critical components of the glomerular filtration barrier, sitting on the outside of the glomerular basement membrane. Primary and secondary foot processes are characteristic for podocytes, but cell processes that develop in culture were not studied much in the past. Moreover, protocols for diverse visualization methods mostly can only be used for one technique, due to differences in fixation, drying and handling. However, we detected by single-cell RNA sequencing (scRNAseq) analysis that cells reveal high variability in genes involved in cell type-specific morphology, even within one cell culture dish, highlighting the need for a compatible protocol that allows measuring the same cell with different methods. Here, we developed a new serial and correlative approach by using a combination of a wide variety of microscopic and spectroscopic techniques in the same cell for a better understanding of podocyte morphology. In detail, the protocol allowed for the sequential analysis of identical cells with light microscopy (LM), Raman spectroscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). Skipping the fixation and drying process, the protocol was also compatible with scanning ion-conductance microscopy (SICM), allowing the determination of podocyte surface topography of nanometer-range in living cells. With the help of nanoGPS Oxyo®, tracking concordant regions of interest of untreated podocytes and podocytes stressed with TGF-β were analyzed with LM, SEM, Raman spectroscopy, AFM and SICM, and revealed significant morphological alterations, including retraction of podocyte process, changes in cell surface morphology and loss of cell-cell contacts, as well as variations in lipid and protein content in TGF-β treated cells. The combination of these consecutive techniques on the same cells provides a comprehensive understanding of podocyte morphology. Additionally, the results can also be used to train automated intelligence networks to predict various outcomes related to podocyte injury in the future.

Effectiveness and durability of anti-graffiti products applied on ETICS: towards a compatible and sustainable graffiti removal protocol

External Thermal Insulation Composite Systems (ETICS) are widely used constructive solutions which aim at enhancing the building thermal performance. Nevertheless, ETICS can often present anomalies (e.g., stains and microcracks) throughout their service life, and vandalism actions, as in the case of graffiti, are rather common in urban areas. The removal of undesired graffiti is generally carried out through invasive chemical-mechanical methods, which may affect the durability of the ETICS. The adoption of anti-graffiti products can be a feasible protection method; however, no comprehensive studies were already addressed on these substrates. This study aims at evaluating the effectiveness, compatibility, and durability of three anti-graffiti products (with permanent, semi-permanent, and sacrificial properties) when applied on different ETICS. The removal of aerosol graffiti paints was carried out with a low-invasive and eco-friendly removal method (i.e., low-pressure steam jet). The water transport properties, as well as color, gloss, and roughness, were evaluated before and after graffiti removal. The durability of the anti-graffiti was also assessed by artificial aging cycles. Results showed that graffiti removal was rather efficient on ETICS with acrylic-based finishing coats and when using (semi) permanent anti-graffiti products (with ΔE*ab < 5, i.e., not macroscopically visible, when comparing cleaned and reference surfaces), although these products can reduce their effectiveness after aging. Conversely, unsatisfactory graffiti cleaning was observed on ETICS with lime-based or silicate-based finishing coats (with ΔE*ab > 5), with considerable alteration also of the water transport properties (reducing water absorption and slowing down the drying kinetic).

Oxidative cyanation of tertiary amines for facile synthesis of tetrahydroisoquinolines with quaternary centers

A facile and efficient synthesis of tetrahydroisoquinolines with quaternary centers has been successfully realized by orchestrating a sequential process consisted of N-bromosuccinimide oxidation and cyanation of tertiary amines. This compatible combination protocol enables one-pot synthesis of α-cyano tetrahydroisoquinolines containing a quaternary center with up to 99% yield.

Imagery perspective in copying movement

Imagery is a cognitive process widely used for learning and performance of motor skills (Morris et al., 2005). In this study, we investigated Internal (IP) and External Imagery Perspectives (EP) and actual copying of movement to deepen understanding of the character of imagery perspectives. Research on IP and EP shows that the superiority of one perspective in terms of performance enhancement is still equivocal. Further research is required to understand the character of IP and EP, and to establish how they affect learning and performance of motor skills. We applied a Stimulus-Response (S-R) compatibility protocol to a finger-movement, choice-reaction time (CRT) task to differentiate response latencies for actual copying of the movement, and IP and EP of the movement, after watching internal and external stimulus videos of the CRT button-pressing task. We used paired t-tests to analyse the differences in decision time and movement time for external and internal stimulus videos, in the physical copying of the movement condition. Analysis of variance (ANOVA) was used to examine differences in response times for movement, IP, and EP imagery conditions for internal and external video stimuli. Results showed that the response times in both imagery perspective conditions were significantly slower than response times in the physical movement condition. It was further revealed that response times were faster for the internal view than the external view for the movement and IP conditions, whereas EP latencies were faster for external than internal video stimuli. We concluded that the S-R compatibility protocol can be used to confirm use of IP and EP, which is questionable in studies where participants are simply instructed to use IP or EP, and it is assumed they follow instructions, or even when they are asked to confirm use of IP or EP at the end of studies.

High-Throughput Cell-Free Screening of Eukaryotic Membrane Proteins in Lipidic Mimetics.

Membrane proteins (MPs) carry out important functions in the metabolism of cells, such as the detection of extracellular activities and the transport of small molecules across the plasma and organelle membranes. Expression of MPs for biochemical, biophysical, and structural analysis is in most cases achieved by overexpression of the desired target in an appropriate host, such as a bacterium. However, overexpression of MPs is usually toxic to the host cells and can lead to aggregation of target protein and to resistance to detergent extraction. An alternative to cell-based MP expression is cell-free (CF), or in vitro, expression. CF expression of MPs has several advantages over cell-based methods, including lack of toxicity issues, no requirement for detergent extraction, and direct incorporation of target proteins in various lipidic mimetics. This article describes a high-throughput method for the expression and purification of eukaryotic membrane proteins used in the author's lab. Basic Protocol 1 describes the selection and cloning of target genes into appropriate vectors for CF expression. Basic Protocol 2 describes the assembly of CF reactions for high-throughput screening. Basic Protocol 3 outlines methods for purification and detection of target proteins. Support Protocols 1-6 describe various accessory procedures: amplification of target, treatment of vectors to prepare them for ligation-independent cloning, and the preparation of S30 extract, T7 RNA polymerase, liposomes, and nanodiscs. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Target selection, construct design, and cloning into pET-based expression vectors Support Protocol 1: Amplification of target DNA Support Protocol 2: Preparation of ligation-independent cloning (LIC)-compatible vectors Basic Protocol 2: Assembly of small-scale cell-free reactions for high-throughput screening Support Protocol 3: Preparation of Escherichia coli S30 extract Support Protocol 4: Preparation of T7 RNA polymerase Support Protocol 5: Preparation of liposomes Support Protocol 6: Preparation of nanodiscs Basic Protocol 3: Purification and detection of cell-free reaction products.

Development of a vendor neutral MRI distortion quality assurance workflow.

With the utilization of magnetic resonance (MR) imaging in radiotherapy increasing, routine quality assurance (QA) of these systems is necessary. The assessment of geometric distortion in images used for radiotherapy treatment planning needs to be quantified and monitored over time. This work presents an adaptable methodology for performing routine QA for systematic MRI geometric distortion. A software tool and compatible protocol (designed to work with any CT and MR compatible phantom on any scanner) were developed to quantify geometric distortion via deformable image registration. The MR image is deformed to the CT, generating a deformation field, which is sampled, quantifying geometric distortion as a function of distance from scanner isocenter. Configurability of the QA tool was tested, and results compared to those provided from commercial solutions. Registration accuracy was investigated by repeating the deformable registration step on the initial deformed MR image to define regions with residual distortions. The geometric distortion of four clinical systems was quantified using the customisable QA method presented. Maximum measured distortions varied from 2.2 to 19.4 mm (image parameter and sampling volume dependent). The workflow was successfully customized for different phantom configurations and volunteer imaging studies. Comparison to a vendor supplied solution showed good agreement in regions where the two procedures were sampling the same imaging volume. On a large field of view phantom across various scanners, the QA tool accurately quantified geometric distortions within 17–22 cm from scanner isocenter. Beyond these regions, the geometric integrity of images in clinical applications should be considered with a higher degree of uncertainty due to increased gradient nonlinearity and B0 inhomogeneity. This tool has been successfully integrated into routine QA of the MRI scanner utilized for radiotherapy within our department. It enables any low susceptibility MR‐CT compatible phantom to quantify the geometric distortion on any MRI scanner with a configurable, user friendly interface for ease of use and consistency in data collection and analysis.

BlueSync: Time Synchronization in Bluetooth Low Energy With Energy-Efficient Calculations

Time synchronization is one of the important services in wireless sensor networks (WSN), and Bluetooth low energy (BLE) is one of the wireless protocols that has drawn a lot of attention recently, with applications in Internet of Things (IoT) and sensor fusion. Although, nonconnectable beacons are the most energy-efficient advertising mode of BLE, no synchronization service is defined for them in the BLE standard. Here, we present <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">BlueSync</i> , a BLE compatible time synchronization protocol, which uses beacons for data transfer. As part of this study, we investigate the tradeoffs between energy efficiency, synchronization error, and speed by considering different drift-management methods and various synchronization packet configurations. Additionally, we propose a Discrete Adjustment technique that can improve energy efficiency in calculating timer ticks updates up to <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$15\times $ </tex-math></inline-formula> without sacrificing accuracy. <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">BlueSync</i> is implemented on two commercial BLE system on chips and tested in 10-min long sessions without any resynchronization. Experimental results show that depending on the settings, <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">BlueSync</i> can achieve average synchronization errors as low as 320 ns per 60 s (single hop) and is the most accurate time synchronization reported on BLE to date.

Generation of Tumor-Specific Cytotoxic T Cells From Blood via InVitro Expansion Using Autologous Dendritic Cells Pulsed With Neoantigen-Coupled Microbeads.

For the past decade, adoptive cell therapy including tumor-infiltrating lymphocytes, genetically modified cytotoxic lymphocytes expressing a chimeric antigen receptor, or a novel T-cell receptor has revolutionized the treatment of many cancers. Progress within exome sequencing and neoantigen prediction technologies provides opportunities for further development of personalized immunotherapies. In this study, we present a novel strategy to deliver in silico predicted neoantigens to autologous dendritic cells (DCs) using paramagnetic beads (EpiTCer beads). DCs pulsed with EpiTCer beads are superior in enriching for healthy donor and patient blood-derived tumor-specific CD8+ T cells compared to DC loaded with whole-tumor lysate or 9mer neoantigen peptides. A dose-dependent effect was observed, with higher EpiTCer bead per DC being favorable. We concluded that CD8+ T cells enriched by DC loaded with EpiTCer beads are tumor specific with limited tumor cross-reactivity and low recognition of autologous non-activated monocytes or CD8+ T cells. Furthermore, tumor specificity and recognition were improved and preserved after additional expansion using our Good Manufacturing Process (GMP)-compatible rapid expansion protocol. Phenotypic analysis of patient-derived EpiTCer DC expanded CD8+ T cells revealed efficient maturation, with high frequencies of central memory and effector memory T cells, similar to those observed in autologous expanded tumor-infiltrating lymphocytes. These results indicate that DC pulsed with EpiTCer beads enrich for a T-cell population with high capacity of tumor recognition and elimination, which are features needed for a T-cell product to be used for personalized adoptive cell therapy.

Understanding Imitation in Papio papio: The Role of Experience and the Presence of a Conspecific Demonstrator.

What factors affect imitation performance? Varying theories of imitation stress the role of experience, but few studies have explicitly tested its role in imitative learning in non-human primates. We tested several predictions regarding the role of experience, conspecific presence, and action compatibility using a stimulus-response compatibility protocol. Nineteen baboons separated into two experimental groups learned to respond by targeting on a touch screen the same stimulus as their neighbor (compatible) or the opposite stimulus (incompatible). They first performed the task with a conspecific demonstrator (social phase) and then a computer demonstrator (ghost phase). After reaching a predetermined success threshold, they were then tested in an opposite compatibility condition (i.e., reversal learning conditions). Seven baboons performed at least two reversals during the social phase, and we found no significant difference between the compatible and incompatible conditions, although we noticed slightly faster response times (RTs) in the compatible condition that disappeared after the first reversal. During the ghost phase, monkeys showed difficulties in learning the incompatible condition, and the compatible condition RTs tended to be slower than during the social phase. Together, these results suggest that (a) there is no strong movement compatibility effect in our task and that (b) the presence of a demonstrator plays a role in eliciting correct responses but is not essential as has been previously shown in human studies.

A Roadmap for the Production of a GMP-Compatible Cell Bank of Allogeneic Bone Marrow-Derived Clonal Mesenchymal Stromal Cells for Cell Therapy Applications.

BackgroundAllogeneic mesenchymal stromal cells (MSCs) have been used extensively in various clinical trials. Nevertheless, there are concerns about their efficacy, attributed mainly to the heterogeneity of the applied populations. Therefore, producing a consistent population of MSCs is crucial to improve their therapeutic efficacy. This study presents a good manufacturing practice (GMP)-compatible and cost-effective protocol for manufacturing, banking, and lot-release of a homogeneous population of human bone marrow-derived clonal MSCs (cMSCs).MethodsHere, cMSCs were isolated based on the subfractionation culturing method. Afterward, isolated clones that could reproduce up to passage three were stored as the seed stock. To select proliferative clones, we used an innovative, cost-effective screening strategy based on lengthy serial passaging. Finally, the selected clones re-cultured from the seed stock to establish the following four-tired cell banking system: initial, master, working, and end of product cell banks (ICB, MCB, WCB, and EoPCB).ResultsThrough a rigorous screening strategy, three clones were selected from a total of 21 clones that were stored during the clonal isolation process. The selected clones met the identity, quality, and safety assessments criteria. The validated clones were stored in the four-tiered cell bank system under GMP conditions, and certificates of analysis were provided for the three-individual ready-to-release batches. Finally, a stability study validated the EoPCB, release, and transport process of the frozen final products.ConclusionCollectively, this study presents a technical and translational overview of a GMP-compatible cMSCs manufacturing technology that could lead to the development of similar products for potential therapeutic applications.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1007/s12015-022-10351-x.