Ginkgo biloba L. exocarp petroleum ether extract inhibits methicillin-resistant Staphylococcus aureus by modulating ion transport, virulence, and biofilm formation in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: As reported in the Ancient Chinese Medicinal Books, Ginkgo biloba L. fruit has been used as a traditional Chinese medicine for the treatment asthma and cough or as a disinfectant. Our previous study demonstrated that G. biloba exocarp extract (GBEE), an extract of a traditional Chinese herb, inhibits the formation of methicillin-resistant Staphylococcus aureus (MRSA) biofilms. However, GBEE is a crude extract that contains many components, and the underlying mechanisms of purified GBEE fractions extracted with solvents of different polarities are unknown. AIM OF THE STUDY: This study aimed to investigate the different components in GBEE fractions extracted with solvents of different polarities and their antibacterial effects and mechanisms against MRSA and Staphylococcus haemolyticus biofilms both in vitro and in vivo. METHODS: The components in different fractions were detected by high-performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS). Microbroth dilution assays and time growth curves were used to determine the antibacterial effects of the fractions on 15 clinical bacterial isolates. Crystal violet staining, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were utilized to identify the fractions that affected bacterial biofilm formation. The potential MRSA targets of the GBEE fraction obtained with petroleum ether (PE), denoted GBEE-PE, were screened by transcriptome sequencing, and the gene expression profile was verified by quantitative polymerase chain reaction (qPCR). RESULTS: HPLC-HRMS analysis revealed that the four GBEE fractions (extracted with petroleum ether, ethyl acetate, n-butanol, and water) contained different ginkgo components, and the antibacterial effects decreased as the polarity of the extraction solvent increased. The antibacterial activity of GBEE-PE was greater than that of the GBEE fraction extracted with ethyl acetate (EA). GBEE-PE improved H. illucens survival and reduced MRSA colonization in model mouse organs. Crystal violet staining and SEM and TEM analyses revealed that GBEE-PE inhibited MRSA and S. haemolyticus biofilm formation. Transcriptional analysis revealed that GBEE-PE inhibits MRSA biofilms by altering ion transport, cell wall metabolism and virulence-related gene expression. In addition, the LO2 cell viability and H. illucens toxicity assay data showed that GBEE-PE at 20 mg/kg was nontoxic. CONCLUSION: The GBEE fractions contained different components, and their antibacterial effects decreased with increases in the polarity of the extraction solvent. GBEE-PE limited MRSA growth and biofilm formation by affecting ion transport, cell wall synthesis, and virulence-related pathways. This research provides a more detailed overview of the mechanism by which GBEE-PE inhibits MRSA both in vitro and in vivo and suggests that GBEE-PE is a new prospective antimicrobial with the potential to be used in MRSA therapeutics in the future.

Polypyrrole-decorated bentonite magnetic nanocomposite: A green approach for adsorption of anionic methyl orange and cationic crystal violet dyes from contaminated water.

In the presented study, a novel polypyrrole-decorated bentonite magnetic nanocomposite (MBnPPy) was synthesized for efficient removal of both anionic methyl orange (MO) and cationic crystal violet (CV) dyes from contaminated water. The synthesis of this novel adsorbent involved a two-step process: the magnetization of bentonite followed by its modification through in-situ chemical polymerization. The adsorbent was characterized by SEM/EDX, TEM/SAED, BET, TGA/DTA-DTG, FTIR, VSM, and XRD studies. The investigation of the adsorption properties of MBnPPy was focused on optimizing various parameters, such as dye concentration, medium pH, dosage, contact time, and temperature. The optimal conditions were established as follows: dye concentration of Co (CV/MO) at 100 mg/L, MBnPPy dosage at 2.0 g/L, equilibrium time set at 105 min for MO and 120 min for CV, medium pH adjusted to 5.0 for MO dye and 8.0 for CV dye, and a constant temperature of 303.15 K. The different kinetic and isotherm models were applied to fit the experimental results, and it was observed that the Pseudo-2nd-order kinetics and Langmuir adsorption isotherm were the best-fitted models. The maximal monolayer adsorption capacities of the adsorbent were found to be 78.74 mg/g and 98.04 mg/g (at 303.15 K) for CV and MO, respectively. The adsorption process for both dyes was exothermic and spontaneous. Furthermore, a reasonably good regeneration ability of MBnPPy (>83.45%/82.65% for CV/MO) was noted for up to 5 adsorption-desorption cycles with little degradation. The advantages of facile synthesis, cost-effectiveness, non-toxicity, strong adsorption capabilities for both anionic and cationic dyes, and easy separability with an external magnetic field make MBnPPy novel.

Think before throw: waste chili stalk powder for facile scavenging of cationic dyes from water.

Chili stalk powder (CS), a non-conventional adsorbent, has been exercised for facile removal of cationic dyes from simulated and wastewater by batch technique. The prepared material has been characterized by Fourier-transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (FESEM), Brunauer-Emmett-Teller analysis (BET), powder X-ray diffraction (powder XRD), and pHZPC and tested best with methylene blue and crystal violet under ambient conditions. FTIR denotes the presence of carbonyl and polyphenolic groups, responsible for dye adsorption. BET surface area analysis evaluates the porous nature and specific surface area of the material, and powder XRD confirms its amorphous nature. The porous structure could be ascertained from the FESEM image, and energy dispersive X-ray analysis (EDX) confirms the elemental composition. The pH above pHzpc shows an increase in removal efficiency. The maximum adsorption capacities are 49.53 and 36.88 mg/g for methylene blue (MB) and crystal violet (CV) respectively. Linear as well as non-linear plots for kinetic and isotherm models were studied. Both dye uptake fits the linear plot of Langmuir adsorption isotherm (R2 = 0.999 and 0.995) and pseudo-second-order kinetics (R2 = 0.998 and 0.999). In the non-linear plot, the adsorption process for both dyes fit Langmuir (R2 = 0.999 for MB and R2 = 0.983 for CV) as well as Freundlich adsorption (R2 = 0.999 for MB and R2 = 0.994 for CV). 75.48% crystal violet (CV) and 73.35% methylene blue (MB) regeneration were successful in 1:1 methanol medium and reused for up to three cycles. The uptake mechanism is suggested to be a union of π-π stacking, electrostatic interaction, and weak hydrogen bonding. The material was tested with industrial effluent to prove its application in real wastewater treatment. Moreover, the material shows superior adsorption capacity than contemporary phytosorbents. To conclude, a zero-cost adsorbent using green chili stalk has been demonstrated for wastewater treatment.

Development and evaluation of a novel water-based pigment marker for radiation therapy skin marking.

Skin marks are widely used in external radiation therapy to ensure the accuracy of the irradiation position. However, conventional skin markers contain harmful substance, so we developed an alternative skin marker. The purpose of this study was to investigate the feasibility of using a novel water-based pigment marker comprising safe materials commonly used in cosmetics for clinical radiation therapy. We investigated various properties of the marker, namely marker longevity, color variety, line visibility, ink bleeding, and line durability, and improved the marker in response to the feel when drawing or being drawn on. The durability of the ink was evaluated by simultaneously applying the new marker and oil-based pen and comparing the period until the marks faded and became invisible. In clinical trial, we applied marks on the skin of 56 patients over three months to observe symptoms and visible changes in the skin. There were no complications of discomfort or pain, owing to the improvements in the marker tip. The marks drawn on the arms of volunteers with the new marker and the oil-based pen remained visible for a mean of 7.2 days and 3.6 days, respectively (P value < 0.001). The percentages of participants with no symptoms and no visible changes were 100%, respectively. We developed an alternative skin marker that complies with current regulatory standards by excluding crystal violet. The newly developed marker has features suitable for clinical use, such as resistance to smudging and water, marker tip shape and texture, and color variations.

Sponge-like biodegradable polypyrrole-modified biopolymers for selective adsorption of basic red 46 and crystal violet dyes from single and binary component systems.

Recently, several researchers have been trying to reduce the ecological effects of water pollution by considering the use of biodegradable materials that prevent the generation of secondary pollution in our environment and enable water reuse. Here, new biodegradable hydrogels based on alginate (Alg), gelatin (Gel) and polypyrrole (PPy) were successfully implemented to remove two known highly toxic cationic dyes from wastewater. The design process was performed in two steps: in-situ polymerization of polypyrrole within the Alg/Gel mixture, followed by hydrogel formation. Biocomposites showed promising efficacy for the removal of both basic red 46 (BR46) and crystal violet (CV) dyes from real and demineralized water samples. However, Alg-Gel-PPy hydrogel showed better selectivity for BR46 than for CV as compared to the pristine Alg-Gel hydrogel. Adsorption of both pollutants on biocomposite hydrogel beads followed the Langmuir isotherm and pseudo-second order kinetic models. Besides, the highest adsorption capacities (125 mg g-1 for BR46 and 88.5 mg g-1 for CV) were obtained for the Alg-Gel-PPy hydrogel, compared with those determined for PPy-free hydrogel (103.09 mg g-1 for BR46 and 86.96 mg g-1 for CV) and remained at a satisfactory level for five adsorption-desorption cycles. Finally, the obtained hydrogels showed excellent biodegradability by natural soil microorganisms, with 91 % decomposition.

The effect and mechanism of iodophors on the adhesion and virulence of Staphylococcus aureus biofilms attached to artificial joint materials.

BACKGROUND: Iodophors are known to be a treatment for biofilm-related periprosthetic joint infection. However, the efficacy and mechanism of eradicating biofilms from different artificial joint materials after iodophor treatment are unknown. This study was conducted to understand the effect and mechanism of iodophors with respect to the adhesion and virulence of Staphylococcus aureus biofilms attached to artificial joint materials. METHODS: Biofilms of Staphylococcus aureus strains were grown on titanium alloy, cobalt chromium molybdenum and polyethylene coupons, which are commonly used materials for artificial joints, for 24 h. Afterward, all coupons were divided into experimental and control groups: (1) exposed to a 0.5 ± 0.05% iodophor for 5 min and (2) exposed to phosphate-buffered saline for 5 min. To gauge the level of biofilm, colony forming units (CFU), live/dead staining confocal microscopy and crystal violet staining were used. Meanwhile, the expression of icaACDR and clfA, which are related to virulence and adhesion, was examined in both the experimental and control groups. RESULTS: A roughly three-log decrease in CFU/cm2 was seen in the viable plate count compared to the control group. Confocal imaging and crystal violet staining verified the CFU data. Moreover, the expression of icaACDR was reduced on three different orthopaedic implant materials, and the expression of clfA was also inhibited on titanium alloy coupons exposed to the iodophor. CONCLUSIONS: Our results indicated that exposure to an iodophor for 5 min could significantly eliminate biofilms. When Staphylococcus aureus that had adhered to these three materials, which were used for artificial joints, was treated with an iodophor for 5 min, the expression of icaACDR was significantly reduced. This provides strong evidence for clinically clearing periprosthetic joint infections without removing the artificial joints.

Catalytic degradation of crystal violet and methyl orange in heterogeneous Fenton-like processes.

Metals-loaded (Fe3+, Cu2+ and Zn2+) activated carbons (M@AC) with different loading ratios (0.1%, 0.5%, 1%, 5% and 10%) were prepared and employed for catalytic degradation of dye model compounds (crystal violet (CV) and methyl orange (MO)) in wastewater by heterogeneous Fenton-like technique. Compared with Cu@AC and Zn@AC, 0.5% Fe3+ loaded AC (0.5Fe@AC) had better catalytic activity for dyes degradation. The effects of dyes initial concentration, catalyst dosage, pH and hydrogen peroxide (H2O2) volume on the catalytic degradation process were investigated. Cyclic performance, stability of 0.5Fe@AC and iron leaching were explored. Degradation kinetics were well fitted to the pseudo-second-order model (Langmuir-Hinshelwood). Almost complete decolorization (99.7%) of 400 mg L-1 CV was achieved after 30 min reaction under the conditions of CV volume (30 mL), catalyst dosage (0.05 g), H2O2 volume (1 mL) and pH (7.7). Decolorization of MO reached 98.2% under the same conditions. The abilities of pyrolysis char (PC) of dyeing sludge (DS) and metal loaded carbon to remove dye pollutants were compared. The intermediate products were analyzed and the possible degradation pathway was proposed. This study provided an insight into catalytic degradation of triphenylmethane- and aromatic azo-based substances, and utilization of sludge char.

One-Pot Synthesis of Silver Nanoparticles from Garcinia gummi-gutta: Characterisation, Antimicrobial, Antioxidant, Anti-Cancerous and Photocatalytic Applications.

BACKGROUND: Methods like the bio-synthesis of silver nanoparticles (Ag NPs) using plant extracts have become promising due to their eco-friendly approach. The study aimed to examine the utilization of Garcinia gummi-gutta fruit phytochemicals as agents in the biosynthesis of Ag NPs, evaluation of the antimicrobial, antioxidant, and anti-cancerous properties, as well as the photocatalytic ability of bio-synthesized Ag NPs against Crystal Violet (CV), a triphenylmethane dye. METHODS: The characterization of the physical properties of the Ag NPs synthesized via the green route was done using UV-Vis spectrophotometry (UV-Vis), X-ray Diffraction (XRD), Fourier Transform Infrared Spectrophotometry (FTIR), Scanning Electron Microscopy (SEM), Zeta potential analysis, and Transmission Electron Microscopy (TEM). The dye degradation efficiency of CV was determined using synthesized Ag NPs under UV light by analyzing the absorption maximum at 579 nm. The antimicrobial efficacy of Ag NPs against E. coli, S. aureus, Candida tropicalis, and Candida albicans was examined using the broth dilution method. The antioxidant and anti-cancer properties of the synthesized Ag NPs were assessed using the DPPH and MTT assays. RESULTS: The UV analysis revealed that the peak of synthesized Ag NPs was 442 nm. Data from FTIR, XRD, Zeta potential, SEM, and TEM analysis confirmed the formation of nanoparticles. The SEM and TEM analysis identified the presence of spherical nanoparticles with an average size of 29.12 nm and 24.18 nm, respectively. Maximum dye degradation efficiency of CV was observed at 90.08% after 320 min without any silver leaching, confirming the photocatalytic activity of Ag NPs. The bio-efficiency of the treatment was assessed using the Allium cepa root growth inhibition test, toxicity analysis on Vigna radiata, and Brine shrimp lethality assay. CONCLUSIONS: The findings revealed the environmentally friendly nature of green Ag NPs over physical/chemically synthesized Ag NPs. The synthesized Ag NPs can effectively be used in biomedical and photocatalytic applications.

Fe/sponge structure peanut shell carbon composite preparation for efficient Fenton oxidation crystal violet.

In order to obtain super synergy effect between adsorption and Fenton oxidation for crystal violet (CV) removement from water, in this study, Fe modified on a sponge structure peanut shell carbon (Fe/SPSC) nanocomposite was successfully synthesized by a wet impregnation method. In the Fe/SPSC sample, the prepared peanut shell carbon had a sponge-like structure, (002) crystal plane of graphite crystallite, and Fe/SPSC composite coexisted Fe2O3 and Fe3O4 crystalline, which could adsorb and enrich crystal violet molecule, decrease the concentration of CV solution rapidly. And also SPSC could do better for electrons transfer and further promote CV oxidation degradation. The removal efficiency results showed that the 7% Fe/SPSC (500 °C, 2 h) had the best CV removal activity. The composite prepared under the optimum conditions is 2.0 g/L, 0.1 mL 30% H2O2, pH = 7.0, 300 mg/L crystal violet water solution, and the CV degradation rate can reach 95.5%, and the CV degradation amount for Fe/SPSC was 143.25 mg/g. It was confirmed that hydroxyl radicals (•OH) is the active center of Fenton oxidation degradation reaction. XPS results showed that Fe, O, and C elements coexist in the 7% Fe/SPSC composite, and N element content increases after the reaction. Remarkable synergies between adsorption and Fenton oxidation, which could make Fe/SPSC, have quick CV abatement ability. The possible systematic effect mechanism of adsorption and Fenton-oxidation CV was also supplied. The present system has advantages on high CV dye degradation performance, no other Fe sludge formation, short reaction time, and better catalyst reusability.

Theranostic approach to specifically targeting the interloop region of BCL2 i-motif DNA by crystal violet.

Ligands that recognise specific i-motif DNAs are helpful in cancer diagnostics and therapeutics, as i-motif formation can cause cancer. Although the loop regions of i-motifs are promising targets for ligands, the interaction between a ligand and the loop regions based on sequence information remains unexplored. Herein, we investigated the loop regions of various i-motif DNAs to determine whether these regions specifically interact with fluorescent ligands. Crystal violet (CV), a triphenylmethane dye, exhibited strong fluorescence with the i-motif derived from the promoter region of the human BCL2 gene in a sequence- and structure-specific manner. Our systematic sequence analysis indicated that CV was bound to the site formed by the first and third loops through inter-loop interactions between the guanine bases present in these loops. As the structural stability of the BCL2 i-motif was unaffected by CV, the local stabilisation of the loops by CV could inhibit the interaction of transcription factors with these loops, repressing the BCL2 expression of MCF-7 cells. Our finding suggests that the loops of the i-motif can act as a novel platform for the specific binding of small molecules; thus, they could be utilised for the theranostics of diseases associated with i-motif DNAs.

A Comparative and Critical Analysis for In Vitro Cytotoxic Evaluation of Magneto-Crystalline Zinc Ferrite Nanoparticles Using MTT, Crystal Violet, LDH, and Apoptosis Assay.

Zinc ferrite nanoparticles (ZFO NPs) are a promising magneto-crystalline platform for nanomedicine-based cancer theranostics. ZFO NPs synthesized using co-precipitation method are characterized using different techniques. UV-visible spectroscopy exhibits absorption peaks specific for ZFO. Raman spectroscopy identifies Raman active, infrared active, and silent vibrational modes while Fourier transforms infrared spectroscopic (FTIR) spectra display IR active modes that confirm the presence of ZFO. X-ray diffraction pattern (XRD) exhibits the crystalline planes of single-phase ZFO with a face-centered cubic structure that coincides with the selected area electron diffraction pattern (SAED). The average particle size according to high-resolution transmission electron microscopy (HR-TEM) is 5.6 nm. X-ray photoelectron spectroscopy (XPS) signals confirm the chemical states of Fe, Zn, and O. A superconducting quantum interference device (SQUID) displays the magnetic response of ZFO NPs, showing a magnetic moment of 45.5 emu/gm at 70 kOe. These ZFO NPs were then employed for comparative cytotoxicity evaluation using MTT, crystal violet, and LDH assays on breast adenocarcinoma epithelial cell (MCF-7), triple-negative breast cancer lines (MDA-MB 231), and human embryonic kidney cell lines (HEK-293). Flow cytometric analysis of all the three cell lines were performed in various concentrations of ZFO NPs for automated cell counting and sorting based on live cells, cells entering in early or late apoptotic phase, as well as in the necrotic phase. This analysis confirmed that ZFO NPs are more cytotoxic towards triple-negative breast cancer cells (MDA-MB-231) as compared to breast adenocarcinoma cells (MCF-7) and normal cell lines (HEK-293), thus corroborating that ZFO can be exploited for cancer therapeutics.

Hydroxyapatite and ionic liquid coupled with hybrid membranes for toxic pollutant removal and remediation.

Access to clean water is the mandatory requirement for every living being to sustain life. So, membrane-based integrated approach of adsorption and separation technology has recently been preferred by scientists over other conventional techniques, for wastewater treatment. Current research focused on the synthesis of novel imidazolium (A1) based IL, which was further functionalized with hydroxyapatite (HAp; extracted from Labeo rohita scales), to create possible solutions towards environmental remediation. Cellulose acetate (CA) was used for the fabrication of three different ionic liquid membranes. All the synthesized products were characterized by FTIR, XRD and TGA. Two dyes of different nature, i.e., congo red (anionic) and crystal violet (cationic) were selected because of their highly toxic and carcinogenic effects, for batch adsorption experiments. Antibacterial activity of the synthesized membranes was also evaluated against S. aureus. Results of the study revealed that CA-HA1 1:2 acted as the best adsorbent towards the removal of crystal violet, exhibiting removal efficiency of 98% with the contact time of 24 h while in case of congo red adsorption, CA-HA1 (1:2) proved as prime adsorbent with the removal efficiency of 96% for the same preceding contact time. Considering the antibacterial character of the synthesized membranes, CA-A1 (1:1) emerged as very efficient antibacterial agent with the inhibition zone of 50 mm after 48 h. The overall behavior of monolayer and multilayer adsorption was witnessed for both dyes while kinetic studies favored the pseudo-second order reaction for all adsorbents.

Precise and Facile Endotracheal Lung-tumor-implantation Mouse Model Visualized by GFP Expression.

BACKGROUND/AIM: In osteosarcoma, lung metastasis is a major cause of cancer-related death, as the 5-year survival rate for patients with metastases is approximately only 20-30%. To develop improved therapeutic strategies against lung-metastatic osteosarcoma, an experimental lung-tumor-implantation mouse model is needed for basic research. In the present study, we developed a precise and facile endotracheal lung-tumor-implantation technique. MATERIALS AND METHODS: For establishment of the lung-tumor-implantation mouse model of metastatic osteosarcoma, 5 mice were used. A 15-mm longitudinal incision was made in the center of the neck to expose the salivary glands. The salivary glands were then split, exposing the trachea covered by the sternohyoid muscles. The trachea was then clearly exposed by cutting the sternohyoid muscles longitudinally. A 22 G gavage needle was tilted slightly toward the left side of the mouse and inserted from the oral cavity into the bronchus, with confirmation of the position of the tip of the gavage needle visualized through the tracheal wall, followed by injection of 0.5% crystal violet to first confirm the accuracy of endotracheal injection in the lung. A 143B-GFP cell suspension (2.0×106 cells/50 µl PBS) was then injected endotracheally in other mice. RESULTS: The procedure, including anesthesia and suturing, took approximately 10 minutes. The left lobe of the lung, in which crystal violet was injected endotracheally, was stained in 3 out of 3 mice (100%). 143B-GFP-osteosarcoma tumors were detected with GFP fluorescence in the left lobe of the lung in 3 out of 4 mice (75%), 5 weeks after endotracheal injection. One mouse died 4 weeks after 143B-GFP-cell implantation. CONCLUSION: This novel technique of establishing tumors in the lung via endotracheal injection of cancer cells is precise and facile and can be used widely, since neither a surgical microscope nor X-ray imaging are needed.

Hydrothermal synthesis of a photocatalyst based on Byrsonima crassifolia and TiO2 for degradation of crystal violet by UV and visible radiation.

This work presents a one-step synthesis methodology for preparing a hydrochar (HC) doped with TiO2 (HC-TiO2) for its application on the degradation of crystal violet (CV) using UV and visible radiation. Byrsonima crassifolia stones were used as precursors along with TiO2 particles. The HC-TiO2 sample was synthesized at 210 °C for 9 h using autogenous pressure. The photocatalyst was characterized to evaluate the TiO2 dispersion, specific surface area, graphitization degree, and band-gap value. Finally, the degradation of CV was investigated by varying the operating conditions of the system, the reuse of the catalyst, and the degradation mechanism. The physicochemical characterization of the HC-TiO2 composite showed good dispersion of TiO2 in the carbonaceous particle. The presence of TiO2 on the hydrochar surface yields a bandgap value of 1.17 eV, enhancing photocatalyst activation with visible radiation. The degradation results evidenced a synergistic effect with both types of radiation due to the hybridized π electrons in the sp2-hybridized structures in the HC surface. The degradation percentages were on average 20% higher using UV radiation than visible radiation under the following conditions: [CV] = 20 mg/L, 1 g/L of photocatalyst load, and pH = 7.0. The reusability experiments demonstrated the feasibility of reusing the HC-TiO2 material up to 5 times with a similar photodegradation percentage. Finally, the results indicated that the HC-TiO2 composite could be considered an efficient material for the photocatalytic treatment of water contaminated with CV.

Inhibition of proliferation, migration, and adhesion of skin fibroblasts by enzymatic poly(gallic acid) grafted with L-Arginine, migration, and adhesion of skin fibroblasts by enzymatic poly(gallic acid) grafted with L-Arginine.

Psoriasis and atopic dermatitis (AD) are characterized by enhanced skin inflammation, which results in hyperproliferation and the recruitment of immune cells into the skin. For that reason, it is needed a chemical capable to reduce cell proliferation and the recruitment of cells. The search for new molecules for therapeutic skin treatment mainly focuses on the antioxidant and anti-inflammatory properties, highlighting the rheological properties of polymeric polypeptides. We studied L-arginine (L-Arg) grafted (-g-) to enzymatic poly(gallic acid) (PGAL). The latter is a multiradical antioxidant with greater properties and thermal stability. The derivative was enzymatically polymerized in an innocuous procedure. The poly(gallic acid)-g-L-Arg molecule (PGAL-g-L-Arg) inhibits bacterial strains which also have been involved in the progression of psoriasis and AD. However, it is important to analyze their biological effect on skin cells. The cell viability was analyzed by calcein/ethidium homodimer assays and crystal violet. The proliferation and cell attachment were determined by a curve of time and quantitation of the optical density of crystal violet. To analyze the cell migration a wound-healing assay was performed. This synthesis demonstrates that it is not cytotoxic at high concentrations (250 µg/mL). We observed a decrease in the proliferation, migration, and adhesion of dermal fibroblasts in vitro but the compound could not avoid the increase of reactive oxygen species in the cell. Based on our findings, PGAL-g-L-Arg is a promising candidate for treating skin diseases such as psoriasis and AD where decreasing the proliferation and cell migration could help to avoid inflammation.

A Histogram-Based Technique for Simultaneous Colorimetric Determination of Malachite Green and Brilliant Green Using Triton X-100 Micelle.

BACKGROUND: Malachite green (MG) and brilliant green (BG) are two synthetic triphenylmethane dyes with applications in the textile and aquaculture industries. They are considered to be environmental contaminants due to their carcinogenic and mutagenic properties. Both dyes have the same bluish-green color in aqueous solutions. OBJECTIVE: The aim of this study is to develop a colorimetric analysis as a very simple and cost-effective method to determine the residues of MG and BG simultaneously in aqueous industrial samples. METHOD: This method is based on the alterations in red, blue, green (RGB) color histograms of the dyes in the presence and absence of Triton X-100 micelle. The images of the samples were taken by a digital camera and converted to the RGB color system using MATLAB software. Partial least-squares regression as a powerful chemometrics tool was used for multivariate calibrations and quantitative measurements. The performance of the proposed method was compared with a simple spectrophotometric method as a reference. RESULTS: Relative errors of prediction for colorimetric and spectrophotometric analysis, respectively, in micellar media were 6.56 and 4.61% for MG and 6.38 and 5.24% for BG. The shortest linear ranges for colorimetric and spectrophotometric analysis, respectively, in micellar media were 0.1-10 and 0.5-5 mg/L for MG and 0.1-15 and 0.5-6 mg/L for BG. The recovery percentages obtained from the analysis of the dyes in real samples of fish-pond water and textile wastewater ranged between 91 and 107%. CONCLUSIONS: The good correlation between the results of the colorimetric analysis and the spectrophotometric analysis indicates the reliability of the proposed colorimetric method. Also, the results of the relative recovery study showed insignificant matrix effect. HIGHLIGHTS: This study demonstrates the ability of the colorimetric analysis coupled with chemometrics tools for simultaneous determination of the analytes even with nearly identical colors.

Isolation, Culture and Morphological Assessment of Primary Cell Lines from Human Primary Oral Squamous Cell Carcinoma Using Explant Technique.

BACKGROUND: Due to many uses of cell culture in cell biology, biotechnology, and medical research, this technique has evolved into a widely used and accepted methodology. The isolation of primary cells from primary cancer tissue is a crucial step in cell culture technology since it offers a trustworthy source for studying the biology, morphology, and molecular evaluation of cancer cells, just like in the oral cavity tissue of patients. Therefore, the technique used for the isolation, culture, and evaluation of these cells is crucial. AIM: The aim of the present study is to isolate and culture the cells from human primary Oral Squamous Cell Carcinoma [OSCC] tissue and evaluate them for morphological variations using an explant method. MATERIALS AND METHODS: The patients with OSCC who were undergoing surgery provided the tissue samples. An explant technique was used to achieve the isolation of cells from tissue samples. Following that, the cells were maintained, subcultured, and stored in accordance with the standard American Type Culture Collection [ATCC] protocol. Routine Hematoxylin & Eosin and crystal violet stains were used. These cells were morphologically studied, and the results were assessed for further studies. RESULTS: We were able to successfully isolate and culture cells from 4 different tissue samples using the explant method. Morphological analysis revealed that one tissue had a significantly distinct presentation of epithelial and stromal cells, whereas the other three tissues had only minor morphological differences predominantly stromal cells. Two tissues were discarded after showing contamination. CONCLUSION: Tissue culture should be done very meticulously specially when oral cavity tissue is used as it is house for millions of microorganisms. The technique must also be thoroughly followed and adjusted accordingly. Using common, inexpensive stains like Hematoxylin and Eosin and crystal violet, which are of great help for examining the morphology of cells routinely.

Iron Porphyrin as a Cytochrome P450 Model for the Degradation of Dye.

Organic dyes are widely used in the textile, biological, medical and other fields. However, a serious environmental problem has appeared because of the presence of organic dyes in industrial aqueous effluents. Thus, the efficient treatment of organic dyes in industrial wastewaters is currently in real demand. The current study investigated the oxidative degradation of the organic dye gentian violet by meso-tetra(carboxyphenyl) porphyriniron(III), [FeIII(TCPP)] as a cytochrome P450 model and iodosylbenzene (PhIO) as an oxidant at room temperature. The degradation reaction was monitored by UV-vis absorption spectroscopy via the observation of UV-vis spectral changes of the gentian violet. The results showed that the efficiency of catalyzed degradation reached more than 90% in 1 h, indicating the remarkable oxidative degradation capacity of the [FeIII(TCPP)]/PhIO system, which provided an efficient approach for the treatment of dyeing wastewater.

Carbon dioxide can inhibit biofilms formation and cellular properties of Shewanella putrefaciens at both 30 °C and 4 °C.

Shewanella putrefaciens (S. putrefaciens), which is a common specific spoilage organism (SSO) of marine fish, has strong spoilage ability even under low-temperature conditions. Carbon dioxide (CO2) was widely applied to control microorganisms in aquatic products package. To explore the regulation mechanism of CO2 on biofilm formation and cell properties of S. putrefaciens, the dynamic formation process of biofilms, cellular surface properties, and cellular metabolic characteristics of S. putrefaciens at both 30 °C and 4 °C in pure CO2 gas were evaluated. As evidenced by the crystal violet staining method, confocal laser scanning microscopy (CLSM) analysis, and field emission scanning electron microscopy (FESEM) observation, dynamic formation process of S. putrefaciens biofilms was apparently delayed by CO2 with integral cellular morphology. The number and viability of sessile cells in S. putrefaciens biofilms was significantly lower than those in normal air composition. The changes in cellular surface properties, such as decreased auto-aggregation and hydrophobicity, might be one of the reasons why biofilms were inhibited by CO2. Inhibition of swimming and swarming motility ability by CO2 could also be observed with significantly shorter bacterial halo diameter. What's more, cellular metabolism was significantly decreased by CO2 according to the results of ATP content, ATPase activity and extracellular proteolytic activity. The influence of CO2 could be both observed whether combined with 30 °C or 4 °C. However, the inhibition produced by CO2 was more pronounced at the incubation temperature of 4 °C. In summary, it could be concluded that the dynamic formation process of S. putrefaciens biofilms and cellular metabolic properties could be inhibited by CO2. This research provided a theoretical basis for better application of CO2 to regulate spoilage microorganisms.