Comprehensive approach to in silico identification and in vitro validation of anti-filarial hit molecules targeting the dimer interface of thioredoxin peroxidase 1 in Wuchereria bancrofti: a progress in anti-filariasis drug development.

Lymphatic filariasis (LF) remains a significant health challenge for populations in developing countries. LF is a parasitic disease transmitted by mosquitoes, mainly caused by the filarial nematode, Wuchereria bancrofti, prevalent in tropical and subtropical regions. Since the present drugs develop complications, including adverse side effects, lack of specificity, and development of drug resistance, the present study focused on developing the potential anti-filariasis drugs targeting crucial proteins for the nematode life cycle. We have identified the therapeutic compounds by targeting the enzyme thioredoxin peroxidase 1 (WbTPx1), which facilitates the conversion of hydrogen peroxide into water, an essential mechanism by which the nematode survives against oxidative stress in the host. This approach might resolve treatment efficacy and activity difficulties at various stages of filarial parasitic worms. We modeled the structure of WbTPx1 and employed the structure-based virtual screening approach, focusing on the dimer interface region of the protein. ADMET prediction profiles of the non-toxic, top-ranked hits with higher docking scores demonstrate higher affinity to the nematode protein than its human homolog. The molecular dynamic simulation studies show WbTPx1-hit complexes' stability and the intactness of hits in the binding site. Further, in vitro validation of identified hits using Setaria digitata, a cattle nematode, showed better IC50 and higher inhibition than the standard drug ivermectin, indicating the potential to inhibit enzyme activity and the development of drug candidates for controlling LF.

Enhancing hydrological insight: isotopic methods revealing groundwater-surface water interactions in the Lower Quang Tri River Group, Vietnam.

The Lower Quang Tri River Group, situated in central Vietnam, faces a myriad of challenges, notably the decline in groundwater levels and the salinisation of both groundwater and surface water, significantly impacting water availability for domestic, agricultural, and industrial purposes. To address these pressing concerns, this study adopts a comprehensive methodology integrating hydrogeological measurements, isotopic techniques, and chemical analyses of various water sources, including local precipitation, surface water bodies, reservoirs, and groundwater samples. Utilising the deuterium and oxygen-18 signatures (δ2H and δ18O) in water molecules as environmental tracers for the assessment of base flow and water sources enables a nuanced understanding of the intricate interaction between surface water and groundwater. Research findings elucidate that during the dry season, groundwater recharge primarily stems from water in the reservoirs over approximately seven months. Base flow contributes between 80 and 85 % of streamflow during the rainy season, escalating to 100 % during the dry season. The mean travelling time of the base flow is estimated at 120 ± 10 days using the sine curve model developed by Rodgers et al. The insights gleaned from this study are poised to play a pivotal role in guiding the local water resources managers in licensing for the exploitation of a right quantities of groundwater as sustainable management strategies in the region.

Ruthenium p-Cymene Complexes Incorporating Substituted Pyridine-Quinoline-Based Ligands: Synthesis, Characterization, and Cytotoxic Properties.

Organometallic complexes of the formula [Ru(N^N)(p-cymene)Cl][X] (N^N = bidentate polypyridyl ligands, p-cymene = 1-methyl-4-(1-methylethyl)-benzene, X = counter anion), are currently studied as possible candidates for the potential treatment of cancer. Searching for new organometallic compounds with good to moderate cytotoxic activities, a series of mononuclear water-soluble ruthenium(II)-arene complexes incorporating substituted pyridine-quinoline ligands, with pending -CH2OH, -CO2H and -CO2Me groups in the 4-position of quinoline ring, were synthesized, for the first time, to study their possible effect to modulate the activity of the ruthenium p-cymene complexes. These include the [Ru(η6-p-cymene)(pqhyme)Cl][X] (X = Cl- (1-Cl), PF6- (1-PF6), pqhyme = 4-hydroxymethyl-2-(pyridin-2-yl)quinoline), [Ru(η6-p-cymene)(pqca)Cl][Cl] ((2-Cl), pqca = 4-carboxy-2-(pyridin-2-yl)quinoline), and [Ru(η6-p-cymene)(pqcame)Cl][X] (X = Cl- (3-Cl), PF6- (3-PF6), pqcame = 4-carboxymethyl-2-(pyridin-2-yl)quinoline) complexes, respectively. Identification of the complexes was based on multinuclear NMR and ATR-IR spectroscopic methods, elemental analysis, conductivity measurements, UV-Vis spectroscopic, and ESI-HRMS techniques. The solid-state structures of 1-PF6 and 3-PF6 have been elucidated by single-crystal X-ray diffraction revealing a three-legged piano stool geometry. This is the first time that the in vitro cytotoxic activities of these complexes are studied. These were conducted in HEK293T (human embryonic kidney cells) and HeLa cells (cervical cancer cells) via the MTT assay. The results show poor in vitro anticancer activities for the HeLa cancer cell lines and 3-Cl proved to be the most potent (IC50 > 80 µΜ). In both cell lines, the cytotoxicity of the ligand precursor pqhyme is significantly higher than that of cisplatin.

Antimicrobial, Antioxidant and Cytotoxic Effects of Essential Oil, Fatty Acids and Bioactive Compounds of Beta vulgaris var. crassa (Fodder Beet).

Beta vulgaris var. crassa is undoubtedly a very important plant that is not used enough in the world. In this study, it was aimed to determine the cytotoxic activities of the components (essential oils, fatty acids, total phenol and flavonoid) found in the leaf parts of Beta vulgaris var. crassa against PC-3, MCF-7 and HeLa cancer cell lines. In addition, the effectiveness of these ingredients against bacteria and fungi that can cause serious health problems in humans was tested. In experiments, three tumor cell lines were exposed to various plant extract concentrations (31.25, 62.5, 125, 250, 500 and 1000 µg/mL) for 72 h. It was found that plant extracts showed high (SI: 2.14 > 2) cytotoxicity to PC-3 cells, moderate (SI: 1.62 < 2) to HeLa cells, and low (SI: 0.93 < 2) cytotoxicity to MCF-7 cells. Also, different plant extract concentrations were found to cause an inhibition rate of 16.3-22.3% in Staphylococcus aureus, 16.8-23.5% in Streptococcus pyogenes and 12-16.2% in Cutibacterium acnes. Similarly, inhibition rates were determined between 9.5-20.7% for Candida albicans, 3.5-7.7% for Candida auris, and 5.5-15.1% for Candida glabrata. The results showed that the plant extract exhibited a concentration-dependent cytotoxic and antimicrobial effect against both cancer cell lines and microbial pathogens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01269-8.

Comparative in-vitro analysis of amniotic Fluid's efficacy in sustaining viability and regulating apoptosis of periodontal fibroblasts versus HBSS.

BACKGROUND: Tooth avulsion necessitates swift replantation, for which the preservation of periodontal ligament (PDL) cell viability is paramount. Various storage media have been explored, yet a comparison between amniotic fluid (AF) obtained at different gestational stages (amniocentesis and full-term) and HBSS is lacking. AIM: This study aims to evaluate AF (amniocentesis and full-term) against HBSS in sustaining PDL cell viability and regulating apoptosis at different time points. MATERIAL AND METHODS: Periodontal fibroblasts cultured in α-MEM were treated with 100% AF (amniocentesis), 100% AF (full-term), and HBSS, incubated for 1, 3, 24, and 48 h at 37°C, and assessed using the MTT assay for viability and AO/EB staining for apoptosis, which was analyzed via fluorescent microscopy after 24 h. Statistical analysis was conducted using one-way ANOVA, multivariate ANOVA, and post hoc Tukey's multiple comparison tests (p < .05). RESULTS: Amniotic fluid (amniocentesis) exhibited the highest optical density (OD), which implies the highest cell viability across time intervals, followed by AF (full-term) and HBSS. While HBSS maintained PDL morphology, both AF groups showed altered morphology. No cell death was observed after 24 h. CONCLUSIONS: Within the limitations of this study, both AF groups showed the potential to sustain PDL cell viability after 1, 3, 24, and 48 h of storage. However, further investigation is warranted regarding their suitability as storage media.

Biological and Physicochemical Analysis of Sr-Doped Hydroxyapatite/Chitosan Composite Layers.

In this work results are presented on the evaluation of HAp, HApSr, HAp_CS, and HApSr_CS layers deposited on Ti substrates regarding L929 cell viability and cytotoxicity as well as antimicrobial activity against Staphylococcus aureus, in connection with their physicochemical properties. The HAp and HApSr layers generated by radio-frequency magnetron sputtering technique were further covered with chitosan by a matrix-assisted pulsed laser evaporation technique. During the plasma depositions, the Ti substrates were heated externally by a home-made oven above 100 °C. The HApSr_CS layers generated on the unpolished Ti substrates at 100 °C and 400 °C showed the highest biocompatibility properties and antimicrobial activity against Staphylococcus aureus. The morphology of the layer surfaces, revealed by scanning electron microscopy, is dependent on substrate temperature and substrate surface roughness. The optically polished surfaces of Ti substrates revealed grain-like and microchannel structure morphologies of the layers deposited at 25 °C substrate temperature and 400 °C, respectively. Chitosan has no major influence on HAp and HApSr layer surface morphologies. X-ray photoelectron spectroscopy indicated the presence of Ca 2p3/2 peak characteristic of the HAp structure even in the case of the HApSr_CS samples generated at a 400 °C substrate temperature. Fourier transform infrared spectroscopy investigations showed shifts in the wavenumber positions of the P-O absorption bands as a function of Sr or chitosan presence in the HAp layers generated at 25, 100, and 400 °C substrate temperatures.

6-Bromo quinazoline derivatives as cytotoxic agents: design, synthesis, molecular docking and MD simulation.

Based on unselectively, several side effects and drug resistance of available anticancer agents, the development and research for novel anticancer agents is necessary. In this study, a new series of quinazoline-4(3H)-one derivatives having a thiol group at position 2 of the quinazoline ring (8a-8 h) were designed and synthesized as potential anticancer agents. The Chemical structures of all compounds were characterized by 1H-NMR, 13C-NMR, and Mass spectroscopy. The antiproliferative activity of all derivatives were determined against two cancer cell lines (MCF-7 and SW480) and one normal cell lines (MRC-5) by the MTT method. Cisplatin, Erlotinib and Doxorubicin were used as positive controls. The results of in vitro screening showed that 8a with an aliphatic linker to SH group was the most potent compound with IC50 values of 15.85 ± 3.32 and 17.85 ± 0.92 µM against MCF-7 and SW480 cell lines, respectively. 8a indicated significantly better potency compared to Erlotinib in the MCF-7 cell line. The cytotoxic results obtained from testing compound 8a on the normal cell line, revealing an IC50 value of 84.20 ± 1.72 µM, provide compelling evidence of its selectivity in distinguishing between tumorigenic and non-tumorigenic cell lines. Structure-activity relationship indicated that the variation in the anticancer activities of quinazoline-4(3H)-one derivatives was affected by different substitutions on the SH position. Molecular docking and MD simulation were carried out for consideration of the binding affinity of compounds against EGFR and EGFR-mutated. The binding energy of compounds 8a and 8c were calculated at -6.7 and - 5.3 kcal.mol- 1, respectively. Compounds 8a and 8c were found to establish hydrogen bonds and some other important interactions with key residue. The DFT analysis was also performed at the B3LYP/6-31 + G(d, p) level for compounds 8a, 8c and Erlotinib. Compound 8a was thermodynamically more stable than 8c. Also, the calculated theoretical and experimental data for the IR spectrum were in agreement. The obtained results delineated that the 8a can be considered an appropriate pharmacophore to develop as an anti-proliferative agent.

Soluble and insoluble expression of recombinant human interleukin-2 protein using pET expression vector in Escherichia coli.

Interleukin-2 has emerged as a potent protein-based drug to treat various cancers, AIDS, and autoimmune diseases. Despite its immense requirement, the production procedures are inefficient to meet the demand. Therefore, efficient production procedures must be adopted to improve protein yield and decrease procedural loss. This study analyzed cytoplasmic and periplasmic IL-2 expression for increased protein yield and significant biological activity. The study is focused on cloning IL-2 into a pET-SUMO and pET-28a vector that expresses IL-2 in soluble form and inclusion bodies, respectively. Both constructs were expressed into different E. coli expression strains, but the periplasmic and cytoplasmic expression of IL-2 was highest in overnight culture in Rosetta 2 (DE3). Therefore, E. coli Rosetta 2 (DE3) was selected for large-scale production and purification. Purified IL-2 was characterized by SDS-PAGE and western blotting, while its biological activity was determined using MTT bioassay. The results depict that the periplasmic and cytoplasmic IL-2 achieved adequate purification, yielding 0.86 and 0.51 mg/mL, respectively, with significant cytotoxic activity of periplasmic and cytoplasmic IL-2. Periplasmic IL-2 has shown better yield and significant biological activity in vitro which describes its attainment of native protein structure and function.

New benzimidazole-oxadiazole derivatives as potent VEGFR-2 inhibitors: Synthesis, anticancer evaluation, and docking study.

We report herein, the design and synthesis of benzimidazole-oxadiazole derivatives as new inhibitors for vascular endothelial growth factor receptor-2 (VEGFR-2). The designed members were assessed for their in vitro anticancer activity against three cancer cell lines and two normal cell lines; A549, MCF-7, PANC-1, hTERT-HPNE and CCD-19Lu. Compounds 4c and 4d were found to be the most effective compounds against three cancer cell lines. Compounds 4c and 4d were then tested for their in vitro VEGFR-2 inhibitory activity, safety profiles, and selectivity indices using the normal hTERT-HPNE and CCD-19Lu cell lines. It was determined that compound 4c was the most effective and safe member of the produced chemical family. Vascular endothelial growth factor A (VEGFA) immunolocalizations of compounds 4c and 4d were evaluated relative to control by VEGFA immunofluorescence staining. Compounds 4c and 4d inhibited VEGFR-2 enzyme with half-maximal inhibitory concentration values of 0.475 ± 0.021 and 0.618 ± 0.028 µM, respectively. Molecular docking of the target compounds was carried out in the active site of VEGFR-2 (Protein Data Bank: 4ASD).

Single-Cell MTT: A Simple and Sensitive Assay for Determining the Viability and Metabolic Activity of Polyploid Giant Cancer Cells (PGCCs).

Solid tumors and tumor-derived cell lines commonly contain highly enlarged (giant) cancer cells that enter a state of transient dormancy (active sleep) after they are formed, but retain viability, secrete growth promoting factors, and exhibit the ability to generate rapidly proliferating progeny with stem cell-like properties. Giant cells with a highly enlarged nucleus or multiple nuclei are often called polyploid giant cancer cells (PGCCs). Although PGCCs constitute only a subset of cells within a solid tumor/tumor-derived cell line, their frequency can increase markedly following exposure to ionizing radiation or chemotherapeutic drugs. In this chapter we outline a simple and yet highly sensitive cell-based assay, called single-cell MTT, that we have optimized for determining the viability and metabolic activity of PGCCs before and after exposure to anticancer agents. The assay measures the ability of individual PGCCs to convert the MTT tetrazolium salt to its water insoluble formazan metabolite. In addition to evaluating PGCCs, this assay is also a powerful tool for determining the viability and metabolic activity of cancer cells undergoing premature senescence following treatment with anticancer agents, as well as for distinguishing dead cancer cells and dying cells (e.g., exhibiting features of apoptosis, ferroptosis, etc.) that have the potential to resume proliferation through a process called anastasis.

Dual function surfactants for pharmaceutical formulations: The case of surface active and antibacterial 1-tolyl alkyl biguanide derivatives.

One interesting field of research in the view of developing novel surfactants for pharmaceutical and cosmetic applications is the design of amphiphiles showing further bioactive properties in addition to those commonly displayed by surface-active compounds. We propose here the chemical synthesis, and characterization of 1-o-tolyl alkyl biguanide derivatives, having different lengths of the hydrocarbon chain (C3, C6, and C10), and showing surface active and antibacterial/disinfectant activities toward both Gram-positive and Gram-negative bacteria. Both surface active properties in terms of critical micelle concentration (CMC) and surface tension at CMC (γCMC), as well as the antimicrobial activity in terms of minimum inhibitory concentrations (MICs), were strongly dependent on the length of the hydrocarbon chain. Particularly, the C6 and C10 derivatives have a good ability to decrease surface tension (γCMC <40 mN/m) at low concentrations (CMC < 12 mM) and a satisfactory antibacterial effect (MIC values between 0.230 and 0.012 mM against S. aureus strains and between 0.910 and 0.190 against P.aeruginosa strains). Interestingly, these compounds showed a disinfectant activity at the tested concentrations that was comparable to that of the reference compound chlorhexidine digluconate. All these results support the possible use of these amphiphilic compounds as antibacterial agents and disinfectants in pharmaceutical or cosmetic formulations.

A Color Indicator Based on 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) and a Biodegradable Poly(ester amide) for Detecting Bacterial Contamination.

Bacterial contamination is a hazard in many industries, including food, pharmaceuticals, and healthcare. The availability of a rapid and simple method for detecting this type of contamination in sterile areas enables immediate intervention to avoid or reduce detrimental effects. Among these methods, colorimetric indicators are becoming increasingly popular due to their affordability, ease of use, and quick visual interpretation of the signal. In this article, a bacterial contamination indicator system was designed by incorporating MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) into an electrospun PADAS matrix, which is a biodegradable poly(ester amide) synthesized from L-alanine, 1,12-dodecanediol, and sebacic acid. Uniaxial stress testing, thermogravimetric analysis and scanning electron microscopy were used to examine the mechanical properties, thermal stability, and morphology of the mats, respectively. The capacity for bacterial detection was not only analyzed with agar and broth assays but also by replicating important environmental conditions. Among the MTT concentrations tested in this study (0.2%, 2%, and 5%), it was found that only with a 2% MTT content the designed system produced a color response visible to the naked eye with optimal intensity, a sensitivity limit of 104 CFU/mL, and 86% cell viability, which showed the great potential for its use to detect bacterial contamination. In summary, by means of the process described in this work, it was possible to obtain a simple, low-cost and fast-response bacterial contamination indicator that can be used in mask filters, air filters, or protective clothing.

Effect of 2-hydroxyethylammonium carboxylate protic ionic liquids on the solubility and cytotoxicity of indomethacin.

Recently, there is a particular interest to utilize protic ionic liquids (PILs) in drug solubility. This study is exploring the effect of three protic ionic liquids (PILs) based on 2-hydroxyethylammonium carboxylate [2-hydroxyethylammonium acetate (MEAA), 2-hydroxyethylammonium lactate (MEAL), and 2-hydroxyethylammonium propionate (MEAP)] on the solubility of the very poorly soluble drug in water, indomethacin (IMC). The shake flask method was used to measure the experimental solubility of IMC at the different temperatures range (298.15-313.15) K. The results demonstrate significantly enhancment the solubility of IMC in PILs compared to pure water, with an approximate increase of 200 times. The experimental solubility data have been correlated using the empirical models which showed the performance as the order: Modified Apelblat-Jouyban-Acree > Van't Hoff-Jouyban-Acree > Modified Apelblat equations and also the performance for the Wilson model indicated as the order (absolute relative deviation): 2-hydroxyethylammonium acetate (3.030) > 2-hydroxyethylammonium propionate (3.239) > 2-hydroxyethylammonium lactate (7.665). Then the thermodynamic dissolution properties were obtained by usage of Gibbs and Van't Hoff equations to investigate the thermodynamic behavior of the IMC in the aqueous solution PILs. Eventually, the cytotoxicity of the co-solvents (PILs) under study was evaluated using a standard MTT assay. The results showed that the cell viability percentage increased in the following order: MEAA < MEAP < MEAL. These findings indicated that these PILs had low to moderate toxicity. It is noteworthy that the functional groups of the anions were not the only determinant factor of the cytotoxicity. Other factors encompassing concentration, exposure time, and cell line characteristics also had significant effects.

Graphene-Functionalized Titanium Carbide Synthesis and Characterization and Its Cytotoxic Effect on Cancer Cell Lines.

Background Graphene is a versatile material with promising applications in various fields such as electronics, energy, biomedicine, and the environment due to its exceptional mechanical strength, thermal and electrical conductivity, transparency, and chemical stability. Graphene has been extensively used in biological and medical settings. MXene is a two-dimensional (2D) material that exhibits a strong affinity for water and electrical conductivity because of its surface terminations (oxygen {-O}, fluorine {-F}, and hydroxyl {-OH}) and transition metal carbide or nitride. MXene has attracted significant attention recently for its wide range of applications and unique properties. This study focuses on the synthesis and characterization of graphene-functionalized MXene. Furthermore, we investigated its cytotoxic effects on cancer cell lines. The characterization of graphene-functionalized MXene is carried out using scanning electron microscopy (SEM), X-ray diffraction(XRD), and Fourier transform infrared spectroscopy (FTIR) assays. Materials and methods Graphene powder was finely ground in isopropyl alcohol and then sonicated for two hours to produce solution A. MXene was synthesized by reacting titanium aluminum carbide (Ti3AlC3) with hydrofluoric acid (HF). A mixture of Ti3AlC3 and HF was heated to 40°C with continuous stirring for 24 hours to form solution B. Subsequently, solutions A and B were combined and stirred for 30 minutes. The resulting mixture was transferred to a hydrothermal reactor and maintained at 180°C for 12 hours. After the completion of the reaction, the resulting material was cooled to room temperature and purified through washing with distilled water, ethanol, and acetone. The sample was then dried at 80°C for 12 hours. Results The X-ray diffraction (XRD) study confirms the formation of graphene-functionalized titanium carbide (Ti3C2). The sharp peaks indicate a highly crystalline nature. Graphene is a sheet-like structure with numerous gaps. Particles exhibit a multitude of voids and pores on their surfaces. Upon incorporation, graphene displays a small sheet-like structure. Graphene-functionalized titanium carbide confirms the presence of distinct layered or sheet-like structures stacked together. Following the addition of the material, some cancer cells are eradicated, and they exhibit increased biocompatibility, demonstrating anticancer activity. Conclusion Graphene-functionalized titanium carbide has been successfully synthesized and characterized, as evidenced by various analytical methods such as X-ray diffraction (XRD), scanning electron microscopy (SEM), and methyl-thiazoldiphenyl-tetrazolium (MTT) assays. The cytotoxic impact of the synthesized graphene-functionalized titanium carbide on cancer cell lines was examined. The findings reveal a notable cytotoxic effect, indicating its potential as an anticancer agent. Further research in collaboration with experts from diverse fields will be crucial to advance and translate this technology into practical applications for cancer patients. Future scope Graphene and titanium carbide are promising materials for cancer research, biomedical applications, and imaging. Nevertheless, additional research is required to comprehend their mechanisms, enhance their properties, assess their safety and efficacy, and conduct clinical trials.

Psidium guajav-mediated zinc oxide nanoparticles as a multifunctional, microbicidal, antioxidant and antiproliferative agent against destructive pathogens.

Bio-inspired zinc oxide nanoparticles are gaining immense interest due to their safety, low cost, biocompatibility, and broad biological properties. In recent years, much research has been focused on plant-based nanoparticles, mainly for their eco-friendly, facile, and non-toxic character. Hence, the current study emphasized a bottom-up synthesis of zinc oxide nanoparticles (ZnO NPs) from Psidium guajava aqueous leaf extract and evaluation of its biological properties. The structural characteristic features of biosynthesized ZnO NPs were confirmed using various analytical methods, such as UV-Vis spectroscopy, X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDX), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), Scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HR-TEM). The synthesized ZnO NPs exhibited a hydrodynamic shape with an average particle size of 11.6-80.2 nm. A significant antimicrobial efficiency with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 40 and 27 µg/ml for Enterococcus faecalis, followed by 30 and 40 µg/ml for Staphylococcus aureus, 20 and 30 µg/ml for Staphylococcus mutans, 30 µg/ml for Candida albicans was observed by ZnO NPs. Additionally, they showed significant breakdown of biofilms of Streptococcus mutans and Candida albicans indicating their future value in drug-resistance research. Furthermore, an excellent dose-dependent activity of antioxidant property was noticed with an IC50 of 9.89 µg/ml. The antiproliferative potential of the ZnO NPs was indicated by the viability of MDA MB 231 cells, which showed a drastic decrease in response to increased concentrations of biosynthesized ZnO NPs. Thus, the present results open up vistas to explore their pharmaceutical potential for the development of targeted anticancer drugs in the future.

Studies on Anticancer Effect of Methanolic Leaf Extract of Justicia gendarussa on Lung Cancer Cell Line.

Objective: The aim of study's goal was to look into the anticancer efficacy of a methanolic extract of Justicia gendarussa against a lung cancer cell line. Materials and Methods: Cell viability assays and cell and nuclear morphology examinations were used to evaluate the anticancer efficacy against methanolic extract of Justicia gendarussa on lung cancer cell lines. The IC50 doses were calculated using different concentrations of Justicia gendarussa extract (0, 10, 20, 40, 60, and 80 µg/mL). Results: The results of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay revealed that the percentage of viability in treated cells was significantly lower as compared with untreated control groups, which represented as 100%, and an inhibitory concentration of 40 µg/mL was observed. Under a phase-contrast microscope, morphological changes revealed cell shrinkage and cytoplasmic membrane blebbing. The apoptotic nuclei (intensely colored, broken nuclei, and compacted chromatin) were examined under a fluorescence microscope. Conclusions: The outcome of the research work on Justicia gendarussa was investigated for anticancer properties. The results revealed the proapoptotic and cytotoxic effects of Justicia gendarussa extract on lung cancer cell lines. From the above results and findings, it could be concluded that the Justicia gendarussa methanolic leaf extract exhibited potent anticancer activity against a lung cancer cell line. Further study needs to be conducted to investigate the active chemicals in the extract as well as the molecular mechanisms underlying its anticancer benefits.

Evaluation of In vitro Anti-Cancer Activity of Methanolic Leaf Extract of Phoenix pusilla on Colon Cancer Cell Line.

Background: Cancer rates continue to climb, owing largely to the world population's aging and growth, as well as economically developing countries, a surge in cancer-causing behavior, particularly smoking. The third or fourth most prevalent type of cancer is colon cancer. Cancer of the large intestine (colon) is one of the primary causes of death from cancer. Colorectal cancer prevention is mostly based on adenomatous disease screening approaches. The cytotoxic and pharmacological properties of Phoenix pusilla are widely documented. As a result, there is little recorded evidence of its cytotoxic activity against colon cancer cells. Therefore, we planned to study the efficacy of a methanolic leaf extract of Phoenix pusilla against in vitro colon cancer cells. Aim: To evaluate the anti-cancer effects of the methanolic leaf extract of Phoenix pusilla on colon cancer cell lines. Materials and Methods: In vitro screening and anti-cancer effects of the methanolic effect of Phoenix pusilla on colon cancer cell lines were assessed by cell viability assays and cell and nuclear morphological studies. For the in vitro cell culture study, different concentrations of Phoenix pusilla leaf extract (0, 25, 50, 75, 100, 150 µg/ml) were used, and IC50 doses were calculated. Results: The results of the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that the fraction of viability cells significantly decreased in treated cells when compared to untreated control groups, was expressed as 100%, and an inhibitory concentration of µg/ml was identified. A phase-contrast microscope was used to observe cell shrinkage and cytoplasmic membrane blebbing. A fluorescent microscope was used to examine the apoptotic nuclei (internally dyed nuclei, shattered nuclei, and condensed chromatin). Conclusion: In conclusion, the present study results showed that the leaf extracts of Phoenix pusilla had a strong cytotoxic effect and induced significant apoptosis in the colon cancer cell lines at a concentration of 75 µg/ml in the 24 h incubation period. More research is needed to investigate the extract's active components as well as the molecular mechanisms underlying its anti-cancer properties.

Cyclopeptide RA-V from Rubia yunnanensis restores activity of Adagrasib against colorectal cancer by reducing the expression of Nrf2.

Adagrasib (MRTX849), an approved and promising KRAS G12C inhibitor, has shown the promising results for treating patients with advanced non-small cell lung cancer (NSCLC) or colorectal cancer (CRC) harboring KRAS-activating mutations. However, emergence of the acquired resistance limits its long-term efficacy and clinical application. Further understanding of the mechanism of the acquired resistance is crucial for developing more new effective therapeutic strategies. Herein, we firstly found a new connection between the acquired resistance to MRTX849 and nuclear factor erythroid 2-related factor 2 (Nrf2). The expression levels of Nrf2 and GLS1 proteins were substantially elevated in different CRC cell lines with the acquired resistance to MRTX849 in comparison with their corresponding parental cell lines. Next, we discovered that RA-V, one of natural cyclopeptides isolated from the roots of Rubia yunnanensis, could restore the response of resistant CRC cells to MRTX849. The results of molecular mechanisms showed that RA-V suppressed Nrf2 protein through the ubiquitin-proteasome-dependent degradation, leading to the induction of oxidative and ER stress, and DNA damage in CRC cell lines. Consequently, RA-V reverses the resistance to MRTX849 by inhibiting the Nrf2/GLS1 axis, which shows the potential for further developing into one of novel adjuvant therapies of MRTX849.

Biosafety measures for Alicyclobacillus spp. strains across various levels of biohazard.

Alicyclobacillus bacteria are important contaminants in the beverage industry because their spores remain in the product after usual pasteurization. At the same time, their impact on human health has yet to be characterized, as it is generally assumed to be low or non-existent. However, these bacteria are causing quality concerns mainly due to odor and taste changes of the product. Since potential health effects are not precisely known, an experimental assessment was performed, including a biosafety assessment of six viable and non-viable vegetative and spore forms of Alicyclobacillus spp. strains using cell cultures and rodent study. The monolayer of Caco-2 (Cancer coli-2) cells was investigated for its adsorption effect on the epithelium of the small intestine of mice. Lactate dehydrogenase leakage (LDH) and transepithelial electrical resistance (TEER) tests were used to ensure the integrity of the cell membrane and tight junctions. The methylthiazole tetrazolium bromide (MTT) assay examined in vitro cytotoxicity in Caco-2 and HepG2 cell lines. The hemolysis of erythrocytes was spectrophotometrically measured. The results showed negligible cytotoxicity or non-toxic response in mice. In conclusion, Alicyclobacillus spp. exhibited biocompatibility with negligible cytotoxicity and minimal safety concerns.

Development of a Convenient and Quantitative Method for Evaluating Photosensitizing Activity Using Thiazolyl Blue Formazan Dye.

Photosensitizers cause oxidative damages in various biological systems under light. In this study, the method for analyzing photosensitizing activity of various dietary and medicinal sources was developed using 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (thiazolyl blue formazan; MTT-F) as a probe. Significant and quantitative decolorization of MTT-F was observed in the presence of photosensitizers used in this study under light but not under dark conditions. The decolorization of MTT-F occurred irradiation time-, light intensity-, and photosensitizer concentration-dependently. The decolorized MTT-F was reversibly reduced by living cells; the LC-MS/MS results indicated the formation of oxidized products with -1 m/z of base peak from MTT-F, suggesting that MTT-F decolorized by photosensitizers was its corresponding tetrazolium. The present results indicate that MTT-F is a reliable probe for the quantitative analysis of photosensitizing activities, and the MTT-F-based method can be an useful tool for screening and evaluating photosensitizing properties of various compounds used in many industrial purposes.