MTS Test Research Articles

The Pruritus Mediator Recombinant Mucunain Induced Apoptosis Through Caspase 8 Signaling Pathway in the Skin Epithelial Cells

Background Mucunain, a cysteine protease extracted from the spicules of cowhage, can induce intense itching as a pruritus mediator independent of histamine. It is well known that erythema, edema and vesicles on skin may occur immediately following contact with Mucunain. A recent study has demonstrated that Mucunain may trigger pruritus and inflammation on the left or right groin of each Maltese–Beagle atopic dog. The aim of this study was to investigate the cytotoxic effect of Mucunain on a cocultured skin epithelial cell model in vitro. Methods Functional recombination Mucunain (rtMucunain) were obtained following induction, purification, and renaturation. Human keratinocytes and dermal fibroblasts were cocultured to construct a cocultured cell model skin epithelium cell model. The growth inhibition was measured using an MTS test. Microscopy was used to observe morphological alterations in cell architecture. The DCFH-DA assay was used to evaluate reactive oxygen species production, the cells death type was identified using a Flow cytometry assay, and the pro-inflammatory cytokines IL-6 and −8 were detected using an ELISA assay. The ability to form an apoptosis network was investigated by a Western blot assay to assess apoptosis-related protein expression. Results Following purification and renaturation, rtMucunain was obtained with a yield of up to 50 mg/L. In the cocultured skin epithelial cells treated with rtMucunain, cell growth was inhibited in a dose- and time-dependent manner, with doses of 2.5 µM or higher resulting in significant cytotoxicity. Morphological analysis revealed increased cell dispersion and abnormal morphology, particularly following prolonged exposure. Levels of reactive oxygen species were elevated, indicating oxidative stress. Flow cytometry and Western-blot data demonstrated that the cells underwent apoptosis, with extrinsic apoptosis pathways (NF-κB and Caspase-8) being activated by rtMucunain. Furthermore, treatment with rtMucunain markedly enhanced levels of IL-6 and IL-8, suggesting a robust pro-inflammatory response. Conclusion rtMucunain showed a strong inflammation and apoptosis effect in a cocultured skin epithelium cell model. This effect may be due to the activated Caspase 8 signaling pathway, on which NF-κB might play an important role. It was the first time to clarify the cytotoxic effect of rtMucunain on the skin epithelial cell model, and the comprehensive understanding the damaged mechanisms of rtMucunain on normal skin cells can provide a data basis for the skin irritated experiment of rtMucunain and its analogues.

P-2171. Comparing Saliva and Nasal Mid-turbinate Swabs for Detection of Respiratory Syncytial Virus (RSV), February 1–April 15, 2024, New Vaccine Surveillance Network

Abstract Background Current diagnostic methods for RSV and other respiratory viruses primarily rely on reverse transcription polymerase chain reaction (RT-PCR) testing of nasopharyngeal (NPS) or nasal mid-turbinate (MTS) swabs. Recent data in adults suggest saliva increases diagnostic yield of RSV. However, the potential of saliva as a specimen type for detecting RSV in children remains understudied. We compared the diagnostic yield of saliva and paired MTS samples for RSV detection in children. Methods Between February 1 to April 15, 2024, a total of 206 paired MTS and saliva samples obtained using the SalivaBio Children’s swab were collected on the same day from subjects under the age of 18 years presenting with acute respiratory illness symptoms. All MTS samples underwent testing using the QIAstat-Dx Respiratory SARS-CoV-2 Panel, which identifies RSV A+B cases. Subsequently, saliva samples from RSV-positive cases identified by MTS testing were extracted using the Kingfisher Apex platform. Positive RSV cases were subtyped by RT-PCR. Results Among 206 paired MTS and saliva specimens tested, RSV A/B was detected in 8 MTS and 7 saliva specimens. Saliva detected RSV A/B in 7 out of 8 (87.5%) samples that tested positive by MTS swabs. Both MTS and saliva tested negative for RSV in 197/206 specimens tested. The median (IQR) Ct values for paired MTS swabs and saliva samples were 25.05 (21.82 – 32.72) and 31.9 (27.45 – 34.48), respectively. The sample missed by saliva had a high Ct value of 36.6. Saliva detected an additional RSV-A positive case with a Ct value of 33.03, which was negative by the MTS swab. Overall concordance of saliva with MTS was 99% (204/206). Conclusion Although the number of RSV detections was low, preliminary findings from this analysis suggest high concordance between MTS and saliva for detection of RSV but that the addition of saliva does not increase diagnostic yield in children. Disclosures Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Grant/Research Support|Abbott: Honoraria|BioMerieux: Grant/Research Support|Cepheid: Grant/Research Support|Diasorin: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Luminex: Grant/Research Support|Qiagen: Grant/Research Support

A new approach to assessing the efficacy of anthelmintic agents in vitro

Aim. To develop a new method to determine the viability of Opisthorchis felineus in vitro using the MTS reagent and to evaluate its applicability for analyzing the efficacy of anthelmintic agents in the treatment of opisthorchiasis.Materials and methods. Golden hamsters were used to create a model of O. felineus infection. The animals were infected with metacercariae obtained from fish of the Cyprinidae family. Three months after infection, adult parasites were extracted from the hepatobiliary system. Their viability was assessed using the motility scale and a new method based on the modified MTS test protocol. To account for differences between the size and number of adult parasite cells, the results were normalized with respect to protein content. To evaluate the feasibility of the new approach in the study of pharmacological activity against opisthorchiasis, the viability of adult parasites in the presence of praziquantel was tested.Results. During incubation of adult flukes in a medium with the addition of the MTS reagent, colored watersoluble formazan was accumulated. Thermal inactivation of parasites significantly decreased the production of this compound. Since the studied adult parasites differed in size and number of cells, the obtained data on their viability were normalized to protein content. The results correlated with the data on parasite viability obtained by the traditional method using the motility scale. Evaluation of praziquantel efficacy at different concentrations using two independent methods (the MTS test and the motility scale) showed that the results of the MTS test were consistent with literature data and comparable with the results obtained using the motility scale.Conclusion. A new method for in vitro evaluation of anti-opisthorchiasis activity of drugs was developed. It is based on the assessment of water-soluble formazan production by adult O. felineus flukes in the culture medium using the MTS reagent for screening anti-opisthorchiasis activity of new anthelmintic drugs.

Evaluation of the Antioxidant Properties and Biological Effects of a Novel Combined Barberry Root-Propolis Extract on HEK293T Cells.

Background/Objectives: The health benefits of honeybee products and herbs are well known, and their appropriate combination may enhance their biological efficacy. This study investigated the biological properties of a combined barberry root and propolis extract (PBE) in comparison to a propolis extract (PE), a barberry root extract (BE), and pure berberine (BN). Methods: The antioxidant properties were evaluated using DPPH and FRAP methods and total phenolic contents (TPC) were assessed by the Folin-Ciocalteu method. HPTLC was used to quantify the BE in the tested samples. Their effect on HEK293T cells was monitored in real-time by using the xCELLigence system which recorded changes in the proliferative activity (PA). The metabolic activity (MA) was evaluated using an MTS test and cell migration was analyzed via a scratch assay. Results: The PE exhibited a higher TPC (198.67 mg/g) than the BE (119.3 mg/g). The PBE exhibited a comparable antioxidant effect to that of the PE. In the cell assays, the PE, the BE, and BN significantly reduced the proliferative activity at higher concentrations (p < 0.0001) while the PBE demonstrated a lower cytotoxicity and proved to be safer for the tested cells. The highest IC50 value was determined for the PBE (130 µg/mL), suggesting that this combination has a reduced cytotoxicity. However, the scratch test did not confirm a significant supportive effect of the PBE on cell migration. Conclusions: Although the PBE did not show enhanced antioxidant properties, it may mitigate cytotoxicity and support proliferation at lower concentrations. This suggests that extraction of raw propolis with a previously prepared barberry extract results in a safer preparation, but its therapeutic potential requires further studies using biological models.

Безразборная диагностика технического состояния модельного силиконового демпфера крутильных колебаний по результатам лабораторных исследований

The results of laboratory studies on the non-dismountable assessment of the technical condition of a model torsional vibration damper conducted at the MTS testing center of the Astrakhan State Technical University are presented. The relevance lies in the practical application of the obtained research results for a non-dismountable assessment of the technical condition of silicone dampers of marine diesel engines and improving the effectiveness of the existing methodology. The assessment is proposed to be carried out using an in-depth analysis of the torsional oscillation function using wavelet analysis and window Fourier transform methods for dynamic modes of operation of a marine power plant with a rapid change in the rotational speed of the shaft line – start, set the rotational speed and stop. These modes can be dangerous for marine propulsion systems, which is confirmed by a number of facts of shaft destruction and loss of propellers during start-up and reverse of the main power plant. As criteria for evaluating the technical condition of the damper are adopted both traditional ones (the magnitude of stresses in the shafts, the amplitude and frequency of torsional vibrations) and the author's ones (the characteristics of the waveform analysis and windowed Fourier transform methods of the torsional vibration function) as well as the magnitude of vibration along three measuring axes. The data were obtained as a result of the use of the author's experimental samples of the torsional vibration monitoring system and a measuring device based on accelerometers in research. As a result of the study, the dependence of the voltage changes in the shafts, the frequency of vibrations, vibration and the characteristics of the waveform of the wavelet analysis of the Morlet function and windowed Fourier transform diagrams of the torsional vibration function on three simulated situations of the technical condition of the silicone damper – during silicone leakage, flywheel mass sticking and normal operation.

PLLA honeycombs activated by plasma and high-energy excimer laser for stem cell support

In this study, we constructed and activated honeycomb structures on perfluorinated substrates subjected to KrF laser treatment (wavelength 248 nm). We selected the biopolymer poly L-lactic acid (PLLA) as the honeycomb material, which was dissolved in a mixture of chloroform/methanol. A micropattern of a plasma-treated perfluorethyleneperopylene (FEP) substrate was prepared by improved phase separation during dip-coating. The PLLA micropattern was subsequently treated with an excimer laser with a laser fluence of 10 mJ.cm-2 and with a different number of laser pulses. Alternatively, plasma exposure can be used as a secondary treatment. The surface morphologies of the pristine and laser-treated PLLA patterns were studied using atomic force and scanning electron microscopy. The surface chemistry was analyzed using energy-dispersive spectroscopy and X-ray photoelectron spectroscopy. In addition, the metabolic activity of adipose stem cells was evaluated using the MTS test, and cell numbers in selected samples were determined. The morphology of cells growing in the honeycomb-like pattern was studied in detail using fluorescence microscopy. In all, we used a combination of honeycomb pattern (HCP) laser treatment and plasma treatment to construct an optimal scaffold for adipose stem cell culture.

Distinct profile of antiviral drugs effects in aortic and pulmonary endothelial cells revealed by high-content microscopy and cell painting assays

Antiretroviral therapy have significantly improved the treatment of viral infections and reduced the associated mortality and morbidity rates. However, highly effective antiretroviral therapy (HAART) may lead to an increased risk of cardiovascular diseases, which could be related to endothelial toxicity. Here, seven antiviral drugs (remdesivir, PF-00835231, ritonavir, lopinavir, efavirenz, zidovudine and abacavir) were characterized against aortic (HAEC) and pulmonary (hLMVEC) endothelial cells, using high-content microscopy.The colourimetric study (MTS test) revealed similar toxicity profiles of all antiviral drugs tested in the concentration range of 1 nM–50 μM in aortic and pulmonary endothelial cells. Conversely, the drugs' effects on morphological parameters were more pronounced in HAECs as compared with hLMVECs. Based on the antiviral drugs' effects on the cytoplasmic and nuclei architecture (analyzed by multiple pre-defined parameters including SER texture and STAR morphology), the studied compounds were classified into five distinct morphological subgroups, each linked to a specific cellular response profile. In relation to morphological subgroup classification, antiviral drugs induced a loss of mitochondrial membrane potential, elevated ROS, changed lipid droplets/lysosomal content, decreased von Willebrand factor expression and micronuclei formation or dysregulated cellular autophagy.In conclusion, based on specific changes in endothelial cytoplasm, nuclei and subcellular morphology, the distinct endothelial response was identified for remdesivir, ritonavir, lopinavir, efavirenz, zidovudine and abacavir treatments. The effects detected in aortic endothelial cells were not detected in pulmonary endothelial cells. Taken together, high-content microscopy has proven to be a robust and informative method for endothelial drug profiling that may prove useful in predicting the organ-specific endothelial toxicity of various drugs.

Polyhydroxylated Spirostanol Saponins from the Rhizomes of Paris dulongensis.

Three new polyhydroxylated spirostanol steroidal saponins, dulongenosides B-D (2-4), along with 14 known compounds, dulongenoside A (1), padelaoside B (5), parisyunnanoside G (6), polyphyllin D (7), ophiopogonin C' (8), formosanin C (9), dioscin (10), paris saponin VII (11), paris H (12), parisyunnanoside I (13), protodioscin (14), proprotogracillin (15), crustecdysone (16), and stigmasterol-3-O-β-d-glucopyranoside (17), were isolated from the rhizomes of Paris dulongensis (Melanthiaceae). Their chemical structures were elucidated based on extensive analyses of NMR and MS data and acidic hydrolyses. The isolates were evaluated for their cytotoxicity to five human cancer cell lines (HL-60, SW480, MDA-MB-231, A549, and A549/Taxol) and the normal human bronchial epithelial cell line BEAS-2B by the MTS test. Compounds 7-12 and 14 showed cytotoxic activity, with IC50 values ranging from 0.20 to 4.35 μM. Proprotogracillin selectively inhibited A549 (IC50=0.58 μM) and A549/Taxol (IC50=0.74 μM) cells, with no significant cytotoxic activity against HL-60, SW480, MDA-MB-231, or BEAS-2B cells, with IC50 values greater than 40 μM.

Investigation of the immunological effects of escitalopram oxalate in the breast cancer co-culture model.

During breast cancer treatment, approximately half of the patients are prescribed psychotropic medication, such as selective serotonin reuptake inhibitors (SSRIs). Escitalopram oxalate is an SSRI used as an antidepressant. In this study, by creating a breast cancer microenvironment with THP-1, MCF-7 and MDA-MB-231 breast cancer co-culture models were created. MCF-7, MDA-MB-231, and THP-1 cell lines to determine the concentration range of the cytotoxic effect of escitalopram oxalate MTS and MTT test were used. IC50 values were determined by the xCELLigence real-time cell analysis (RTCA) system. Apoptotic activities and cytokine levels were determined by flow cytometry. In the xCELLigence real-time analysis made according to the results, the IC50 value of escitalopram oxalate was measured as 13.7 μM for MCF-7 and 10.9 μM for MDA-MB-231. The IC50 value was measured as 54.6 μM for MCF-7 and 58.4 μM for MDA-MB-231 in xCELLigence analysis with tamoxifen. According to the MTS test results, the IC50 value of tamoxifen for THP-1 was 92.03 μM and the IC50 value for escitalopram oxalate was 95.32 μM. In the co-culture model, the immunological effects of escitalopram oxalate on MCF-7 cells were 2.8%, 11.1%, 15.6%, 10.6%, and 12.1% for interleukin (IL)-1β, IL-6, IL-8, IL-10, and TNF-α, respectively, while MDA effects on MB-231 cells, respectively, were 2.1%, 15.9%, 16.2%, 8.8%, and 11.8%. According to the results obtained, it was concluded that the immunological effects of escitalopram oxalate are more effective than tamoxifen and that it can be used as an adjunctive agent in breast cancer treatment.

Mechanical strength change and coal damage analysis of frozen saturated bitumite after cryogenic freezing

The changes in mechanical strength and damage analysis of bitumite under the action of liquid nitrogen (LN2) freezing are critical issues that need to be addressed in the application of LN2 in coal seam permeability enhancement. Using an MTS testing machine, strain gauges, and ultrasonic detection instruments, experiments were conducted to obtain stress-strain curves and ultrasonic time-domain images of coal samples after single freezing and cyclic freeze-thaw processes. These data were analyzed to understand the mechanical performance and internal fracture evolution of coal samples subjected to ultra-low temperatures of LN2. The experimental results indicate that: (1) the uniaxial compressive strength and elastic modulus of coal samples subjected to a single LN2 freeze are positively correlated with the freezing time from 0 to 50 min, and negatively correlated with the freezing time from 50 to 180 min. (2) The uniaxial compressive strength and elastic modulus of coal samples subjected to cyclic freezing and thawing with LN2 decrease as the number of freeze-thaw cycles increases. (3) The Poisson's ratio of coal samples subjected to single freezing and cyclic freezing-thawing is negatively correlated with the absolute freezing time. Additionally, the decrease in Poisson's ratio is greater in coal samples subjected to cyclic freezing-thawing compared to those subjected to single freezing. (4) As the absolute freezing time increases, the ultrasonic waveforms of the coal samples begin to become disordered, which is manifested by a decrease in amplitude and a delay in the arrival time of the first wave. Under ultra-low temperature conditions, LN2 can deteriorate the mechanical properties of coal samples, with the degree of deterioration being greater under cyclic freezing-thawing than single freezing. This study can provide theoretical guidance for increasing the permeability of coal seams.

Failure analysis and heat treatment process optimization of NOS525 rotary flat binaural hot forging die

A NOS525 steel rotary flat binaural hot forging die frequently failed too early due to collapse and wear in service. In this paper, the rotating planar binaural failure die was taken as the research object. Firstly, the sample design and sample processing of the failure die with different depths and directions were carried out. The material composition, hardness and metallographic structure of the samples were analyzed, and the tensile and impact properties were tested by direct-reading spectrometer, Rockwell hardness tester, optical microscope, scanning electron microscope, MTS testing machine and impact testing machine, etc. Meanwhile, with the help of Deform-HT software and JMatPro software, the coupling simulation of hardness, temperature and structure of the heat treatment of NOS525 steel was realized, and the hardness change of the material after heat treatment was predicted and verified by heat treatment test. The research results show that the composition of the die material is abnormal, especially the excessive P and C elements, at the same time, there is a phenomenon of quenching overheating and insufficient tempering, resulting in insufficient strength, toughness and hardness of the die material, in addition, the die surface is subjected to the cold and hot alternating load, and finally, the die surface collapses and wear too soon, resulting in its failure. Through the combination of heat treatment simulation optimization and test, the optimal heat treatment temperature of NOS525 steel was finally determined, that is, the quenching temperature of 1090℃, the first tempering temperature of 560℃, the second tempering temperature of 600℃, the third tempering temperature of 600℃, so that the mechanical properties of the material have been significantly improved.

Effects of heat treatment on tensile properties of 3D-printed short fiber-reinforced composites

3D-printed carbon fibre-reinforced composites (CFRC) using fused deposition modelling (FDM) offer the potential for building complex geometries and low waste. However, these composites have weaker interlaminar bonding and higher void content than traditional composites. This paper explores the effect of temperature on improving the mechanical properties of 3D-printed short carbon fibre- reinforced polyamide (PACF). Two types of printed structures were tested: one with a 0° build orientation (parallel to extruder movement) and the other with a 90° build orientation (perpendicular to extruder movement). Both samples were heat-treated at 150°C. Force vs. displacement data was obtained from the MTS testing machine. After that, the tested samples were viewed under an optical microscope. Images obtained from the optical microscope are then analyzed to see how the samples failed by checking the microstructure. The average ultimate strength, ultimate strain, and elastic modulus were used for the analysis. The sample follows a trend where the strength and elastic modulus increase after heat treatment. The result also showed that the 0° build orientation samples have higher mechanical performance than the 90° build orientation sample. Also, from the ultimate strain values, it was evident that samples printed in the 0° absorbed more energy, exhibiting 83% higher resistance before final failure. Lastly, an optical microscope was used to investigate the failure mechanisms of the samples.

Cancer Drug Resistance: Targeting Proliferation or Programmed Cell Death.

The development of resistance to chemotherapy is one of the main problems for effective cancer treatment. Drug resistance may result from disturbances in two important physiological processes-cell proliferation and cell death. Importantly, both processes characterize alterations in cell metabolism, the level of which is often measured using MTT/MTS assays. To examine resistance to chemotherapy, different cancer cell lines are usually used for the in vitro modulation of developing resistance. However, after the creation of resistant cell lines, researchers often have difficulty in starting investigations of the mechanisms of insensitivity. In the first stage, researchers should address the question of whether the drug resistance results from a depression of cell proliferation or an inhibition of cell death. To simplify the choice of research strategy, we have suggested a combination of different approaches which reveal the actual mechanism. This combination includes rapid and high-throughput methods such as the MTS test, the LIVE/DEAD assay, real-time cell metabolic analysis, and Western blotting. To create chemoresistant tumor cells, we used four different cancer cell lines of various origins and utilized the most clinically relevant pulse-selection approach. Applying a set of methodological approaches, we demonstrated that three of them were more capable of modulating proliferation to avoid the cytostatic effects of anti-cancer drugs. At the same time, one of the studied cell lines developed resistance to cell death, overcoming the cytotoxic action.

Fatigue Studies of Welded Stainless Steel 304L: Effect of Manufacturability and Weld Type, Choice of Fatigue Model, and Comparison to Pristine Material

Abstract This study investigates fatigue life and fracture in welded stainless steel 304L (SS 304L) resulting from load-controlled fatigue. This was done for specimens from two welding manufacturers. Mechanical testing was carried out using a MTS test frame in uniaxial tension. Specimens fabricated in 3 weld configurations (butt, lap, butt-with-backer) were cycled at stress ratio R = 0.1, frequency f = 5 Hz, and experienced varying mean tensile stress. A comparison of the fatigue life of welded and unwelded (or pristine) specimens determined that the fatigue life of welded specimens is considerably shorter. The microstructure in the parent material (PM), heat affected (HAZ) and fusion zones (FZ) was analyzed using SEM, and results suggest that, for a limited number of specimens (backer welds), fracture in the (FZ) was primarily the result of Zener-Stroh cracks. Most welded specimens fractured at the weld-HAZ interface (weld toe), and we postulate that stress concentrations primarily due to abrupt geometrical change at the weld interface led to crack initiation there. Additional stress concentrations at welding induced defects (resulting from high temperature) such as second phase particles, fatigue induced defects, and pre-existing impurities are responsible for crack initiation, crack growth, and eventual fracture. Comparison of unwelded and welded SN curve data using numerical curve fitting methods reveal that the fatigue life of pristine SS 304L follows the expected sigmoid shape found in literature (specifically the Chandran fatigue model), while that of welded SS 304L deviates considerably in low cycle fatigue (LCF) and follows the Basquin power law equation.

Assessment of the Possibility of Validating the Durability Testing Method for Large-Sized Vehicles Based on Changes in Shock Absorber Characteristics—Preliminary Tests

The use of road simulators in vehicle durability tests enables a significant reduction in time related to the implementation of a new product. However, there is still a need to improve the methods of these tests in order to obtain a full correlation between the time of the tests on the simulator with respect to the mileage of the vehicle in real road conditions. The aim of this work is a validation and improvement of the durability testing method for large-sized vehicles. Comparative tests of shock absorbers dismantled from semi-trailers with different mileage and shock absorbers dismantled from semi-trailers after durability tests conducted at the MTS test bench are presented. The relationship between the maximum damping force and the displacement speed of the shock absorber piston was examined. Additionally, the influence of the shock absorber temperature on the damping force was analyzed. As a result of the tests, the dynamic characteristics of the shock absorbers were determined and the relationship between the maximum damping force and the mileage of the trailers was determined. It was found that the load on the trailers during their operation has a significant impact on the change in the characteristics of the shock absorbers. What is new in this publication is the validation of the methodology for conducting durability tests previously proposed by the authors based on the selected component, which is the shock absorber. The authors have not yet encountered in the literature any attempt to validate the methodology for conducting durability tests for large-sized vehicles.

Thiazole Derivatives as Modulators of GluA2 AMPA Receptors: Potent Allosteric Effects and Neuroprotective Potential.

Thiazole carboxamide derivatives were synthesized in this investigation, with a subsequent examination of their impact on GluA2 AMPA receptors. The synthesized compounds, namely MMH-1-5, were subjected to characterization using high-resolution mass spectrometry (HRMS), proton nuclear magnetic resonance (1H-NMR), and carbon-13 nuclear magnetic resonance (13C-NMR). The present work thoroughly investigates the impact of five thiazole derivatives on GluA2 AMPA receptors. This investigation examined their effects on both whole-cell currents and receptor kinetics. In addition, the cytotoxicity of the samples was assessed using the MTS test. The compound MMH-5 had the highest effect level, resulting in a notable drop in current amplitude by a factor of six. Similarly, MMH-4 and MMH-3 also caused major reductions in the current amplitude. The compounds mentioned above also influenced the rates of deactivation and desensitization. MMH-5 and MMH-4 exhibited an increase in deactivation, while MMH-5 showed reduced desensitization. Our research findings highlight the efficacy of MMH-5 as a negative allosteric modulator of GluA2 AMPA receptors, exerting substantial effects on both the magnitude and time course of receptor activity. Significantly, the compound MMH-2 demonstrated noteworthy cytotoxic effects, as evidenced by cell viability rates dropping below 6.79% for all cancer cell lines and 17.52% for the normal cell line (LX-2). Of particular interest is the pronounced cytotoxicity observed in MMH-5, suggesting its potential as a safe neuroprotective agent targeting the AMPA receptor, as indicated by cell viability percentages exceeding 85.44% across all cancer and normal cell lines. Docking simulations were performed to determine possible modes of interaction between MMH5 and the GluA2-AMPA receptor (PDB:7RZ5). The abovementioned facts and the well-documented effects of further thiazole derivatives provide a strong foundation for future research endeavors to enhance tailored treatments for neurological disorders that rely heavily on GluA2 signaling. The present study elucidates the intricate association between thiazole derivatives and GluA2 receptors, providing valuable perspectives on the prospects of enhanced and specific therapeutic interventions for diverse neurological conditions.

Heterologous Expression of Codon-Optimized Azurin Transferred by Magnetofection Method in MCF-10A Cells.

Transfection efficiency of the immortalized human breast epithelial cell line MCF-10A remains an issue that needs to be resolved. In this study, it was aimed to deliver a recombinant DNA (pCMV-Azu-GFP) to the MCF-10A cells by the magnetofection method using magnetic nanoparticles (MNPs) and a simple magnet to accelerate the DNA delivery. Surface positively modified silica-coated iron oxide MNPs (MSNP-NH2) were produced and characterized via TEM, FTIR, and DLS analyses. The recombinant DNA (rDNA) was obtained by the integration of codon-optimized azurin to produce a fusion protein. Then, rDNA cloned in Escherichia coli cells was validated by sequence analysis. The electrostatically conjugated rDNA on MSNP-NH2 with an enhancer polyethyleneimine (PEI) was studied by agarose gel electrophoresis and the optimum conditions were determined to apply to the cell. A dose-dependent statistical difference was observed on treated cells based on the MTS test. The expression of the fusion protein after magnetofection was determined using laser scanning confocal microscope imaging and western blot analysis. It was observed that the azurin gene could be transferred to MCF-10A cells by magnetofection. Thus, when the azurin gene is used as a breast cancer treatment agent, it can be expressed in healthy cells without toxic effects.

Therapeutic effects of guanidine hydrochloride on breast cancer through targeting KCNG1 gene

BackgroundTriple-negative breast cancer (TNBC) is one of the subtypes of breast cancer (BC) that is associated with poor survival rates and failure to respond to hormonal and targeted therapies. ObjectiveThe aim of this study was to identify a specific gene at the expression level for TNBC and targeting of this type of breast cancer based on it. Using TCGA database, genes that are particularly high expression in TNBC subtypes compared to other BC subtypes (in terms of receptor status) and normal samples were identified and their sensitivity and specificity were evaluated. Using PharmacoGX and Drug Bank data, drug sensitivity and drug-appropriate genes were identified, respectively. The effects of the identified drug on triple-negative cell lines (MDA-MB-468) were evaluated in comparison with the cell line of other subtypes (MCF7) by apoptosis and MTS tests. ResultsData analyzes showed that the expression level of KCNG1 gene in the TNBC subgroup was significantly higher compared to other BC subtypes from the KCN gene family and ROC results showed that this gene had highest sensitivity and specificity in TNBC subtype. The results of drug resistance and sensitivity showed that an increase in the expression level of KCNG1 was associated with sensitivity to Cisplatin and Oxaliplatin. Moreover, Drug Bank results showed that Guanidine hydrochloride (GuHCl) was a suitable inhibitor for KCNG1. In vitro results showed that the expression level of KCNG1 was higher in MDA-MB-468 compared to MCF7. In addition, the rate of apoptosis in response to GuHCl treatment in MDA-MB-468 cell line as TNBC cell model was higher than MCF7 in the same concentration. ConclusionThis study revealed that GuHCl could be a suitable treatment for TNBC subtype by targeting of KCNG1.

Poly(vinyl alcohol)/Silk Fibroin/Ag-NPs Composite Nanofibers as a Substrate for MG-63 Cells' Growth.

Nanofiber scaffolds of polyvinyl alcohol, silk fibroin from Bombyx mori cocoons, and silver nanoparticles were developed as a substrate for MG-63 growth. The fiber morphology, mechanical properties, thermal degradation, chemical composition, and water contact angle were investigated. In vitro tests were performed by the cell viability MTS test of MG-63 cells on electrospun PVA scaffolds, mineralization was analyzed by alizarin red, and the alkaline phosphatase (ALP) assay was evaluated. At higher PVA concentrations, Young's modulus (E) increased. The addition of fibroin and silver nanoparticles improved the thermal stability of PVA scaffolds. FTIR spectra indicated characteristic absorption peaks related to the chemical structures of PVA, fibroin, and Ag-NPs, demonstrating good interactions between them. The contact angle of the PVA scaffolds decreased with the incorporation of fibroin and showed hydrophilic characteristics. In all concentrations, MG-63 cells on PVA/fibroin/Ag-NPs scaffolds had higher cell viability than PVA pristine. On day ten of culture, PVA18/SF/Ag-NPs showed the highest mineralization, observed by the alizarin red test. PVA10/SF/Ag-NPs presented the highest alkaline phosphatase activity after an incubation time of 37 h. The achievements indicate the potential of the nanofibers of PVA18/SF/Ag-NPs as a possible substitute for bone tissue engineering (BTE).

Effect of Commonly Used Cosmetic Preservatives on Healthy Human Skin Cells

Cosmetic products contain preservatives to prevent microbial growth. The various types of preservatives present in skincare products applied on the skin induce many side effects. We tested several types of preservatives such as phenoxyethanol, methyl paraben, propyl paraben, imidazolidinyl urea (IU), the composition of gluconolactone and sodium benzoate (GSB), diazolidinyl urea (DU), and two grapefruit essential oils, one of which was industrially produced and a second which was freshly distilled from fresh grapefruit peels. This study aimed to find the relationship between preservative concentration, cell growth, collagen secretion, and cell viability. We hypothesized that these products induced a decrease in collagen secretion from human dermal fibroblasts. Our research, for the first time, addressed the overall effect of other preservatives on skin extracellular matrix (ECM) by studying their effect on metalloproteinase-2 (MMP-2) activity. Except for cytotoxicity and contact sensitivity tests, there are no studies of their effect on skin ECM in the available literature. These studies show potential antimicrobial activity, especially from the compounds IU and DU towards reference bacteria and the compounds methyl paraben and propyl paraben against reference fungi. The MTS test showed that fibroblasts are more sensitive to the tested group of preservatives than keratinocytes, which could be caused by the differences between the cells' structures. The grapefruit oils exhibited the most cytotoxicity to both tested cell lines compared to all considered preservatives. The most destructive influence of preservatives on collagen synthesis was observed in the case of IU and DU. In this case, the homemade grapefruit oil turned out to be the mildest one. The results from a diverse group of preservatives show that whether they are natural or synthesized compounds, they require controlled use. Appropriate dosages and evaluation of preservative efficacy should not be the only aspects considered. The complex effect of preservatives on skin processes and cytotoxicity is an important topic for modern people.