Plate Adhesion Research Articles

Microcavity-assisted cloning (MAC) of hard-to-clone HepG2 cell lines: cloning made easy

Cloning is a key molecular biology procedure for obtaining a genetically homogenous population of organisms or cell lines. It requires the expansion of new cell populations starting from single genetically modified cells. Despite the technical progress, cloning of many cell lines remains difficult. Cloning often fails either due to the strenuous conditions associated with manipulating cells or because many cells don’t tolerate a single-cell state. Here we describe a new cloning method utilizing low adhesion microcavity plates. This new technique, named microcavity-assisted cloning (MAC) was developed to clone difficult-to-clone HepG2 cells. The clones were produced following CRISPR/Cas9 knockout of the GSTA1 gene by a random distribution of 200, 400, and 800 cells into 550 microcavities of a 24-well low adhesion plate originally designed for the culture of spheroids. The knockout of GSTA1 was verified at the protein level using Western blotting. The advantages of the MAC method are its low cost, ease of the procedure, and the possibility of scaling up the throughput and automatization.

Demonstration of potential DNA contamination introduced by laboratory consumables using Fluorescein

The development of increasingly efficient DNA extraction and profiling kits has increased the amount of allelic information obtained from trace DNA samples, but also inadvertently, increased the detection of DNA contamination. This study aimed to evaluate the potential of DNA transfer using fluorescein, fluorescent under an alternate light source, in the use of a range of forensically relevant DNA profiling consumables. An evaluation of two pre-lysis methods adopting three different sample tubes, some with deliberate seal damage, showed the PrepFiler™ Automated Forensic DNA Extraction Kit caused leakage and crusting when the rim of the PrepFiler™ LySep column was compromised, but no leakage was observed under the same conditions using the Investigator STAR Lyse&Prep kit. The AutoLys tube showed minimal leakage using the PrepFiler™ chemistry. A DNA extract tube with an external thread, similar to the AutoLys tube, showed no leakage after fridge or freezer storage. However, it highlighted that a centrifugal spin does not guarantee all the DNA will pool at the base of the tube. A comparison of adhesive plate sealing films to 8-well strip caps for sealing 96-well PCR plates showed the adhesive plate sealing films presented a lower risk of DNA transfer, largely due to the adhesion of dispersed liquid on the sticky surface of the film. Overall, this study highlighted a number of variables that may be considered in the development of more refined contamination minimisation protocols in respect to increased sensitivities of DNA profiling.

Stability impact on wind turbine blades trailing edge from bonding zone modelling methods

Abstract The trailing edge of wind turbine blade will undergo a large deformation under the action of load, resulting in a higher risk of damage and failure. The finite element analysis modeling method directly affects the simulation result, furthermore impacts the result of blade structure design. Therefore, it is necessary to find out a modeling method, which is capable to reflect the stability of bonding regions more accurately. This paper aims to discuss the influences of different finite element modeling methods on the buckling stability of blade trailing edge. The influences of shell element, tetrahedron element and hexahedron element on the blade stiffness are considered. The bonding region were simulated separately by combining shell element with bonding web, shell element with tetrahedron element and shell element with hexahedron. The influences of different bonding region modeling methods on the trailing edge buckling factor were analyzed by using the eigenvalue buckling analysis method of finite element. The simulation results provided data support for the future validation of the blade trailing edge bonding area modeling method. The results show that, the tetrahedral mesh provides a higher stiffness, followed by the hexahedral mesh, and the adhesive web form has the smallest stiffness. In order to simulate the more realistic stiffness of the adhesive area at the trailing edge of the blade, adhesive web plates can be built in the flange area of the blade root and the core material area of the trial adhesive mold; The direct bonding area can be simulated employing the hexahedron mesh; The tetrahedral mesh has high stiffness and is not suitable for modeling the adhesive area at the trailing edge. Blade simulation modeling can flexibly use different adhesive forms according to actual situations, ensuring the reliability of the simulation model.

3D Modeling: Insights into the Metabolic Reprogramming of Cholangiocarcinoma Cells.

Developing accurate in vitro models that replicate the in vivo tumor environment is essential for advancing cancer research and therapeutic development. Traditional 2D cell cultures often fail to capture the complex structural and functional heterogeneity of tumors, limiting the translational relevance of findings. In contrast, 3D culture systems, such as spheroids, provide a more physiologically relevant context by replicating key aspects of the tumor microenvironment. This study aimed to compare the metabolism of three intrahepatic cholangiocarcinoma cell lines in 2D and 3D cultures to identify metabolic shifts associated with spheroid formation. Cells were cultured in 2D on adhesion plates and in 3D using ultra-low attachment plates. Metabolic exchange rates were measured using NMR, and intracellular metabolites were analyzed using LC-MS. Significant metabolic differences were observed between 2D and 3D cultures, with notable changes in central carbon and glutathione metabolism in 3D spheroids. The results suggest that 3D cultures, which more closely mimic the in vivo environment, may offer a more accurate platform for cancer research and drug testing.

Hygrothermal effect of bio-inspired helicoid laminate plate for strengthening damaged RC beam

This study investigates the influence of moisture and Thermo-hygroscopic influence on bio-inspired helicoidally laminated composite plates (BHLC) for strengthening damaged reinforced concrete (RC) beams. Borrowing inspiration from nature’s design principles, BHLC plates represent a novel approach to rehabilitation. An analytical approach based on deformation compatibility is employed, assuming constant shear and normal stresses across the adhesive layer within the framework of linear elasticity. BHLC plates are implemented to restrict crack propagation within the RC beam. The analytical methodology considers equilibrium and deformation compatibility across all components: the concrete beam, BHLC plate, and adhesive layer. A parametric study explores the sensitivity of interfacial stresses to parameters such as laminate and adhesive stiffness, plate thickness, and helicoidal layup configuration. The results reveal a significant influence of these factors on the magnitude of maximum interfacial shear and normal stresses. This study establishes a foundation for future analyses of damaged RC beams strengthened with BHLC plates, potentially leading to advancements in rehabilitation methodologies and improved crack propagation modeling in RC beams.

Flexural Behavior Evaluation for Seismic, Durability and Structure Performance Improvement of Aged Bridge According to Reinforcement Methods

Among infrastructure, concrete bridges are the most exposed to various environmental effects. Structural degradation occurs due to natural and artificial influences shortening the lifespan of the structure. Therefore, bridges need to be reinforced over time. The structures used in this study are re-formed using aged bridge floor decks that have been used for 50 years, approximately. The fiber-reinforced polymer (FRP) adhesion method, using sheets and plate forms, was selected among various reinforcement methods to investigate the performance of reinforced structures. We have tested various reinforcement scenarios including one and two layers FRP sheets and FRP plates. The mechanical properties of the reinforced structures were evaluated experimentally through tensile strength and flexural test experiments. In contrast to most available literature focused on model-based studies, our present work represents an experimental test validation of structural reinforcement on an actual bridge. Our results indicate that fiber-based reinforcement in sheet form exhibits higher performances of the reinforced structure compared to reinforcement using the plate form. This study is intended to provide sufficient data for reinforcing bridge floors that could be used for reference at future construction sites.

Stress wave propagation law in CFRP under laser impact conditions

The paper analyzes the propagation law of stress waves in CFRP unidirectional plate specimens and adhesive plate specimens under laser shock conditions using a combination of experimentation and simulation. The finite element model is corrected by using the back particle velocity curve obtained from the experiment. Subsequently, the spatiotemporal stress distribution is extracted from the finite element model. The results indicate an overall attenuation trend of the stress wave propagating in the unidirectional plate. In the adhesive plate, the peak value of the stress wave increases first at the adhesive layer position due to the acoustic impedance mismatch and energy absorption characteristics of the adhesive layer material and CFRP. Then it tends to increase after decay. Moreover, the behavior characteristics of the stress wave differ significantly when it propagates from high-impedance material to low-impedance material compared to when it propagates from low-impedance material to high-impedance material.

Fabrication of High-Performance Insulated Metal Substrates Employing h-BN Mixture/Epoxy Composite Coated on Roughened Copper Plate

In this work, an Insulated Metal Substrate (IMS) for packaging high-power electronic devices was created using hexagonal Boron Nitride (h-BN) mixture-based Thermally Conductive Adhesive (TCA) and copper plate with a matte side. Mechanical exfoliation of micron-scale BN particles results in a BN mixture containing h-BN nanosheets, micron particles, and nanoparticles. TCA was synthesized by dispersing a 40 % BN mixture modified by γ-aminopropyltriethoxysilane (KH550) into epoxy resin. The TCA has a relatively high thermal conductivity of 4.48 W·m-1·K-1 and a breakdown strength of 15.32 kV·mm-1. As the TCA film was coated on the matte side of the copper plate, the peel strength can reach 1.63 N·mm-1, indicating excellent practicality in the field of electronic packaging.

Effect of UV-C disinfection and copper plating on healthcare-associated infections in a NICU with high ESBL infections

IntroductionHealthcare-associated infections (HCAIs) in neonates are frequent and highly lethal, in particular those caused by extended spectrum beta-lactamase (ESBL) producing bacteria. We evaluated the beneficial effects of ultraviolet C (UV-C) disinfection and copper adhesive plating on HCAIs in the Neonatal Intensive Care Unit (NICU) of a third level paediatric hospital in Mexico City, both in combination of hand-hygiene (HH) and prevention bundles. MethodsAll NICU patients were included. There were 4 periods (P): P1: HH monitoring and prevention bundles; P2: P1+UV-C disinfection; P3: P2+Copper adhesive plating on frequent-contact surfaces and P4: Monitoring of P3 actions. Results552 neonates were monitored during 15,467 patient days (PD). HCAI rates decreased from 11.03/1000 PD in P1 to 5.35/1000 PD in P4 (p=0.006). HCAIs with bacterial isolates dropped from 5.39/1000 PD in PI to 1.79/1000 PD in P4 (p=0.011). UV-C and copper were associated with significant HCAI prevention (RR 0.49, CI95% 0.30–0.81, p=0.005) and with lesser HCAIs with bacterial isolates (RR 0.33, CI95% 0.14–0.77, p=0.011). ConclusionsCopper adhesive plating combined with UV-C disinfection were associated with a drop in HCAI rates and with the elimination of ESBL-caused HCAIs. Hence, we propose that these strategies be considered in MDRO proliferation preventions.

Experimental Study on Repairing/Restoration and Reinforcement Methods of the Reinforced Concrete Structures Damaged by Earthquakes

For earthquake-damaged reinforced concrete structures, the static loading test method is adopted to carry out loading tests under cyclic loading on reinforced concrete beams with different reinforcement rates and columns with different axial compression ratios, and the effect of reinforcing and repairing the damaged reinforced concrete structure by using adhesive steel plates and carbon fiber cloth is investigated. Through comparative studies of structural hysteresis curves and skeleton curves under different reinforcement methods, it is concluded that reinforcement has significantly improved the hysteresis characteristics and ductility of members, and the seismic performance of the carbon fiber reinforcement method is better than that of the steel plate reinforcement. This study provides valuable references and suggestions for practical earthquake-damaged building reinforcement and repair works and seismic reinforcement works.

Study on Chemical Activity of Electroless Copper Plating

Electroless copper plating of insulating substrates typically requires the pre-deposition of Pd/Sn catalyst layer onto the surface to initiate the chemical reactions; Accordingly, it’s high-profile process to achieve efficient Pd/Sn cluster adhesion and good copper plating performance. This paper showed copper thickness and dimple hole depth of the electroless plating were affected by chemicals following different activation times. The composition, microstructure, size and roughness evolution were characterized using Transmission Electron Microscope (TEM). The structural studies revealed that copper particle size depends not only on the size and coverage of the Pd/Sn cluster, but also on the degree of activation of the chemicals. In order to make deposition copper more compact and uniform to decrease dimple hole depth, dummy boards are generally used to enhance the potion activity before electroless copper plating. This paper will provide chemical activity and plating efficiency relationship to avoid poor activity impact plating quality and process event.

Staphylococcus capitis Bloodstream Isolates: Investigation of Clonal Relationship, Resistance Profile, Virulence and Biofilm Formation.

Staphylococcus capitis has been recognized as a relevant opportunistic pathogen, particularly its persistence in neonatal ICUs around the world. Therefore, the aim of this study was to describe the epidemiological profile of clinical isolates of S. capitis and to characterize the factors involved in the persistence and pathogenesis of these strains isolated from blood cultures collected in a hospital in the interior of the state of São Paulo, Brazil. A total of 141 S. capitis strains were submitted to detection of the mecA gene and SCCmec typing by multiplex PCR. Genes involved in biofilm production and genes encoding enterotoxins and hemolysins were detected by conventional PCR. Biofilm formation was evaluated by the polystyrene plate adherence test and phenotypic resistance was investigated by the disk diffusion method. Finally, pulsed-field gel electrophoresis (PFGE) was used to analyze the clonal relationship between isolates. The mecA gene was detected in 99 (70.2%) isolates, with this percentage reaching 100% in the neonatal ICU. SCCmec type III was the most prevalent type, detected in 31 (31.3%) isolates and co-occurrence of SCCmec was also observed. In vitro biofilm formation was detected in 46 (32.6%) isolates but was not correlated with the presence of the ica operon genes. Furthermore, biofilm production in ICU isolates was favored by hyperosmotic conditions, which are common in ICUs because of the frequent parenteral nutrition. Analysis of the clonal relationship between the isolates investigated in the present study confirms a homogeneous profile of S. capitis and the persistence of clones that are prevalent in the neonatal ICU and disseminated across the hospital. This study highlights the adaptation of isolates to specific hospital environments and their high clonality.

Experimental and numerical investigation on failure behavior of hybrid bonded/bolted GFRP single-lap joints under static shear loading

The mechanical performance of the GFRP (Glass Fiber Reinforced Polymer) hybrid bonded/bolted single-lap joints under static shear loading is investigated. Static loading tests considering the hybrid joints with one bolt, four bolts, and nine bolts were first conducted to study the failure mechanism of hybrid joints, where failure process, rigidity, failure load, and failure modes were focused. Finite element (FE) analysis involving the material degradation and failure laws was also carried out for exploring the internal mechanism of joints’ failure. Based on the verified FE model, the effects of plate thickness and bolt edge distance on the failure capacity and strength of the hybrid joints were further investigated by performing a numerical parametric analysis. Results show that, the failure process of the hybrid joints can be divided into two stages which correspond to the failures of adhesive layer and GFRP plates, respectively. The joint rigidity of joints increases by 98.9 %∼133.1 % comparing the second loading stage to the first stage, and it is found that the rigidity in the first stage is hardly affected by the number of fasten bolts, while it decreases with more bolts in the second stage due to the severe incompleteness of GFRP plates. The failure patterns of the hybrid joints mainly include the slippage between connected plates, the compressive failure and shear failure of GFRP material, and the inclination of bolts. A proper arrangement of bolts can lead to shear failure of GFRP which makes full use of the material property. The final failure loads of four-bolt and nine-bolt joints are respectively 14.5 % and 22.9 % higher than the single-bolt specimen. As the plate thickness getting greater, the shear bearing capacity of hybrid joint increases almost linearly, while it would increase first and then descend with increasing bolt edge distance. In general, the achievement of this work can provide references for the design and further study of the GFRP hybrid joints.

Novel CDK4/6 lnhibitor, WXJ-8 -Exerts Anti-effects Through Cell Cycle in Breast Cancer

The novel Abemaciclib derivative WXJ-8 may be a promising anti-tumour drug; however, its mechanism of action and clinical value need to be further verified in research. Cell cycle regulating systems are critical in parthenogenesis and progression. Because of its involvement in regulating cell cycle checkpoints, the CDK4/6-CyclinD1-Rb-E2F1 signalling pathway has been proposed as a possible therapeutic target for breast cancer. As a result, WXJ-8, a new and highly specific CDK4/6 inhibitor, was developed and synthesized in this investigation. The activity of WXJ-8 against breast cancer cells in vitro was detected by MTT method. The main subjects were MDA-MB-231 and MCF-7 cells. Plate cloning, wound healing, invasion and adhesion experiments revealed that WXJ-8 had strong anti-proliferation, migration, invasion, and adhesion effects on breast cancer cells. Protein blotting revealed significant down regulation of CDK4, CDK6, p-Rb, E2F1 and other cell cycle factors in the CDK4/6-CyclinD1-Rb-E2F1 pathway, as well as activation of Bcl-2, Caspase-3, and Cleaved Caspase-3 apoptotic factor expression, resulting in MDA-MB-231 cell cycle arrest and apoptosis. Finally, our research implies that WXJ-8 might be a potential anti-triple-negative breast cancer medication candidate.

Artificial intelligence-based analysis of time-lapse images of sphere formation and process of plate adhesion and spread of pancreatic cancer cells.

Background: Most pancreatic cancers are pancreatic ductal adenocarcinomas (PDAC). Spherical morphology formed in three-dimensional (3D) cultures and the effects of anticancer drugs differ between epithelial and mesenchymal PDAC cell lines. In the human pancreas, cancer cells form 3D tumors, migrate to adjacent tissues, and metastasize to other organs. However, no effective methods exist to examine the ability of the tumor mass to migrate to surrounding tissues in vitro. We used spheres formed in 3D culture to investigate whether the migratory ability of tumors of PDAC cell lines, including epithelial and mesenchymal cell lines, varies. Methods: Sphere formation and adhesion and spread on culture plates were examined by artificial intelligence-based analysis of time-lapse imaging using five epithelial and three mesenchymal PDAC cell lines. Fused and non-fused areas of the sphere surface during sphere formation on low-attachment plates, the adhesion area to normal culture plates, and the sphere area maintaining its original form during adhesion to plates were measured. Results: Immunocytochemical staining confirmed that E-cadherin was highly expressed in epithelial PDAC spheres, as was vimentin in mesenchymal PDAC spheres, in 2D culture. When forming spheres using low-attachment plates, most epithelial PDAC cell lines initially showed decreased sphere area, and then the covering cells fused to form a smooth surface on the sphere. Mesenchymal PANC-1 and MIA PaCa-2 cells showed little reduction in sphere area and few areas of sphere surface fusion. When formed PDAC spheres were seeded onto normal culture plates, spheres of epithelial PK-8 cells-which have the highest E-cadherin expression, form numerous cysts, and have smooth sphere surfaces-did not adhere to normal plates even after 60h, and epithelial PK45-P and T3M-4 spheres hardly adhered. Conversely, the area of adhesion and spread of mesenchymal PANC-1 and KP4 cell spheres on normal plates markedly increased from early on, forming large areas of attachment to plates. Conclusion: Seeding spheres formed in 3D culture onto culture plates can clarify differences in tumor migration potential to surrounding areas. The masses formed by each PDAC cell line varied in migratory ability, with mesenchymal PDAC masses being more migratory than epithelial PDAC masses.

Comparison of Mechanical Properties of Three Tissue Conditioners: An Evaluation In Vitro Study.

Introduction: Tissue conditioners have been widely used in various clinical applications in dentistry, such as treating inflamed alveolar ridges, temporarily relining partial and complete dentures, and the acquisition of functional impressions for denture fabrication. This study aimed to investigate the mechanical properties of the most prevalent tissue conditioner materials on the market, including Tissue Conditioners (TC), Visco Gel (VG), and FITT (F). Materials and Methods: The three tissue conditioners, TC, VG, and F, were assessed based on the parameters mentioned above. The following tests were performed based on the ISO 10139-1 and ISO 10139-2 requirements: Shore A hardness, denture plate adhesion, sorption, water solubility, and contraction after 1 and 3 days in water. Additional tests are described in the literature, such as ethanol content and gelling time. The tests were carried out by storing the materials in water at 37 °C for 7 days. Results: The gel times of all tested materials exceeded 5 min (TC = 300 [s], VG = 350 [s]). In vitro, phthalate-free materials exhibited higher dissolution in water after 14 days (VG = -260.78 ± 11.31 µg/mm2) compared to F (-76.12 ± 7.11 µg/mm2) and experienced faster hardening when stored in distilled water (F = 33.4 ± 0.30 Sh. A, VG = 59.2 ± 0.60 Sh. A). They also showed greater contractions. The connection of all materials to the prosthesis plate was consistent at 0.11 MPa. The highest counterbalance after 3 days was observed in TC = 3.53 ± 1.12%. Conclusions: Materials containing plasticizers that are not phthalates have worse mechanical properties than products containing these substances. Since phthalates are not allowed to be used indefinitely in medical devices, additional research is necessary, especially in vivo, to develop safe materials with superior functional properties to newer-generation alternatives. In vitro results often do not agree fully with those of in vivo outcomes.

Nonsurgical attachment possibilities for bone conduction hearing aids : Comparison of coupling using external pressure versus an adhesive plate

Bone conduction hearing systems are used for patients with conductive or combined hearing loss who cannot be adequately treated with air conduction hearing aids or surgery. These hearing systems can be surgically implanted or reversibly attached using bone conduction eyeglasses or a rigid or soft headband. Anonsurgical and pressure-free alternative is fixation via an adhesive plate. The aim of this study was to compare the energy transferred from the hearing aid to the mastoid when attached via anew adhesive plate versus via asoft headband. In addition, the comfort and the durability of the adhesive plate was evaluated. Atotal of 30subjects were tested. The transferred energy was measured by accelerometer, which recorded the sound energy at the maxillary teeth. Wearing comfort, fixation duration (until plate becomes loose), and the skin reaction were examined using aquestionnaire after the subjects had worn the adhesive plate with and without hearing aid for amaximum of 7days. The skin reaction was also assessed clinically. There was asignificant difference in the transferred energy in favor of the soft headband at 0.5, 1, and 2 kHz. On the other hand, there was ahigh level of satisfaction and acceptance regarding the aesthetics and the wearing time of the adhesive plate, without skin irritations. The difference in the transferred energy up to 2 kHz is probably due to the lack of pressure from the adhesive plate. This could possibly be compensated after appropriate adjustment of the speech processor. Based on the positive properties of the adhesive plate in terms of comfort, it could be used as an alternative to the soft headband.

Analysis of the Mechanical Performance and Durability of Adhesively Bonded Joints Used in the Milling Tool Industry

Epoxy adhesives, widely used in multiple structural applications, are used in the milling tool industry to replace brazing and mechanical fastening when joining the cutting bits to the tool body; though their durability is still a concern. This work aims to evaluate and characterise the effect of environmental factors associated with a tool’s life cycle on the performance of these bonded joints. A gravimetric analysis was conducted on bulk adhesive plates for water, cutting emulsion and dielectric fluid to obtain diffusion and relaxation rates. Novel real joint shear specimens were developed to enable strength testing on joints which are comparable with the final application. These specimens were immersed in fluids and subjected to thermal cycles or a corrosive finishing surface treatment to simulate the tool’s life cycle. The joint’s resistance was then benchmarked. Lastly, a dimensional variation test was carried out on tool prototypes before and after ageing, showing no significant dimensional variation which could compromise the cutting performance of the tool. Overall, though it was possible to identify a decrease in strength of around 20% in most tests, joint strength was still within the values necessary for safe operation, with a large safety factor still being retained.

Abstract 1749: The synergic effect of cisplatin and a chemokine receptor antagonist sensitizes drug resistant gastric cancer cells and inhibits tumoroid formation

Abstract Introduction: Gastric cancer (GC) is an important cause of death worldwide. Cisplatin (CDDP) is the most used drug in the chemotherapy of advanced GC. However, CDDP resistance reduces GC survival. The CCR5/CCL5 axis has been associated with GC development, but its role in CDDP resistance in GC has not been elucidated. The aim of this study was to determine the effects of CCR5/CCL5 axis blockade by a chemokine receptor antagonist (CRA), alone and in combination with CDDP, on in vitro and 3D tumorid models from AGS R-CDDP cells. Methodology: AGS R-CDDP cells (CDDP-resistant human gastric adenocarcinoma cells) were previously established using a stepwise drug dosing protocol and characterized. CCL5 was selected through transcriptomic analysis and its expression level was validated by qRT-PCR. Cytotoxicity assays were determined by using MTT. Apoptosis and cell cycle assays were evaluated by flow cytometry. Tumoroid formation was performed from AGS R-CDDP cells in low adhesion plates (NunclonTMSpheraTM). After 10 days in culture, tumoroid >150 µm in diameter were scored and pharmacological stimuli were added on days 14, 17 and 20. CRA was used alone and in combination with CDDP in all assays. Results: CRA/CDDP combination triggered a re-sensitized phenotype in AGS R-CDDP cells, decreasing cell viability but not increasing apoptosis compared to independent treatments. AGS R-CDDP cells treated with CRA/CDDP were mainly arrested in S phase. CCL5 showed a decrease in mRNA levels after the CRA/CDDP combination compared to separate treatments. AGS R-CDDP cells showed a higher potential in the ability to form tumoroids compared to AGS WT cells. Finally, CRA/CDDP combination inhibited tumoroid formation compared to independent treatments. Discussion and Conclusion: CCL5 showed a decrease in mRNA levels after the CRA/CDDP combination, possibly correlating with allosteric inhibition of the CCR5 receptor. CRA/CDDP combination has a synergic effect that sensitizes AGS R-CDDP cells and inhibits tumoroid formation. This combination could be used as a potential coadjuvant in GC therapy allowing to reduce the doses of CDDP and therefore to reduce side effects. Citation Format: Bárbara Mora-Lagos, María Elena Reyes, Lorena Lobos-Gonzalez, Matías del Campo, Carmen Gloria Ili, Kurt Buchegger, Yuselin Mora, Louise Zanella, Ismael Riquelme, Priscilla Brebi. The synergic effect of cisplatin and a chemokine receptor antagonist sensitizes drug resistant gastric cancer cells and inhibits tumoroid formation [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 1749.

Chemical reprogramming of human embryonic fibroblasts into neural progenitor cells in vitro

To establish a protocol for reprogramming human embryonic fibroblasts (HEFs) into chemically induced neural progenitor cells (ciNPCs). In the two-staged reprogramming of HEFs, the intermediate compact cell colonies were first chemically induced in KSR medium containing small-molecule compounds (VCR) for 15 days in normoxia, followed by the lineage-specific induction stage, in which the compact cell colonies were digested with 0.25% trypsin and the cells were cultured in low adhesion plates. After formation of a large number of free-floating neurospheres 2 days later, the ciNPCs were labeled with CM-DiI and transplanted into rat models of Parkinson's disease (PD)to observe the survival, migration and differentiation of the cells in PD brain. After induction with VCR for 10 days under normoxic condition, compact cell colonies occurred in HEF cultures (approximately 40 colonies in each well containing 1×105 HEFs), and most of the colonies expressed high levels of alkaline phosphatase. A large number of free-floating neurospheres formed 2 days after passage and were defined as P1 ciNPCs. These ciNPCs exhibited typical neurosphere-like structures and expressed NPC-specific markers (nestin, Sox2, and Pax6). Under neuronal or glial differentiation condition, the ciNPCs expressed the neuron-specific marker Tuj1 and the astrocyte-specific marker GFAP. These ciNPCs could differentiate into Tuj1+, GFAP+, TH+ and GABA+ cells 4 weeks after transplantation into the brain of PD rats. HEFs can be directly reprogrammed into ciNPCs using smallmolecule compounds without the need of introducing exogenous genes. This success may provide a solution to the shortage of donor materials for neuroscience research and treatment of neurodegenerative diseases.