Number Of Viable Cells Research Articles

Ethanolic extract from leaves of tithonia diversifolia induces apoptosis in HCT-116 cells through oxidative stress

ABSTRACT Tithonia diversifolia is a perennial bushy plant found in South America with significant ethnopharmacological importance as an antimalarial, antidiabetic, antibacterial, and anticancer agent. The aim of the present study was to determine the cytotoxicity of the ethanolic extract from leaves of T. diversifolia (TdE) on human cancer cell lines (HCT-116, SNB-19, NCIH-460 and MCF-7), as well as the mechanism of action involved in cell death and cellular modulation of oxidative stress. The TdE exhibited significant activity with IC50 values ranging from 7.12 to 38.41 μg/ml, with HCT-116 being the most sensitive cell line. Subsequent experiments were conducted with HCT-116 cell line. TdE decreased the number of viable cells, followed by induction of apoptotic events, increase in mitochondrial membrane permeabilization, and enhanced G2/M phase of the cell cycle. Pro-oxidative effects including elevated acidic vesicular organelle formation, lipid peroxidation, and nitric oxide by-products, as well as reduced levels of intracellular glutathione and reactive oxygen species production were also observed following incubation with TdE, which may lead to DNA damage followed by apoptotic cell death. These results demonstrate the potential of TdE ethanolic leaf extraction for biological activity and enhance the importance of continuing to study natural sources of plants for the development of anticancer agents.

Evaluation of the Potential Protective Effect of Ellagic Acid against Heavy Metal (Cadmium, Mercury, and Lead) Toxicity in SH-SY5Y Neuroblastoma Cells

Ellagic acid (EA), a polyphenolic constituent of plant origin, has been thoroughly investigated for its hypothesised pharmacological properties among which antioxidant and neuroprotective activities are included. The present study was designed to explore whether EA could attenuate heavy metal (cadmium, mercury, and lead)-induced neurotoxicity in SH-SY5Y cells, which were utilized as a model system for brain cells. MTT and LDH assays were performed to examine the viability of the SH-SY5Y cells after exposure to Cd, Hg, and Pb (either individually or in combination with EA) as well as the effects of necrotic cell death, respectively. Furthermore, 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), a cell-based assay, was performed to determine whether EA could protect SH-SY5Y from heavy metal-induced oxidative stress. Results allowed us to assess the capability of EA to enhance the number of viable SH-SY5Y cells after exposure to heavy metal toxicity. Pre-treatment with EA showed a considerable, concentration-dependent, cytoprotective effect, particularly against Cd2+-induced toxicity. This effect was confirmed through the reduction of LDH release after the simultaneous cell treatment with Cd2+ and EA compared with Cd2+-treated cells. Furthermore, a significant, concentration-dependent decrease in reactive oxygen species (ROS) production, induced by H2O2 or heavy metals, was observed in the same model. Overall, the obtained results provide further insight into the protective role of EA against heavy metal-induced neurotoxicity and oxidative stress, thus indicating the potential beneficial effects of the consumption of EA-rich foods. However, to confirm its effects, well-designed human randomized controlled trials are needed to fill the existing gap between experimental and clinical research.

Comparison of the Antibacterial Efficacy of Bamboo Shoot Ethanol Extract With Chlorhexidine Mouth Rinse Against Salivary Streptococcus mutans and Lactobacillus acidophilus: An Ex Vivo Study.

Background Dental caries is the most prevalent polymicrobial oral infectious disease tormenting individuals' healthy lifestyles and presents a significant public health problem. The objective of this study was to evaluate and compare the antibacterial properties of different concentrations of bamboo shoot ethanol extract with chlorhexidine mouth rinse on isolates of salivary Streptococcus mutans (S. mutans) and Lactobacillus acidophilus (L. acidophilus). Materials and methods Non-stimulated salivary samples from 30 young adults were treated ex vivo with bamboo shoot ethanolic extract at concentrations of 30 µg/ml, 40 µg/ml, 50 µg/ml, and 60 µg/ml. The colony-forming units were quantified by measuring the number of viable bacterial cells. Inhibition zones were evaluated using the agar diffusion method. One-way ANOVA and post-hoc test were used to analyze the significant difference between variables using SPSS version 22.0 (IBM Corp., Armonk, NY). Results The mean zone of inhibition with bamboo shoot ethanolic extract against salivary S. mutans (23.00 ± 0.816) and L. acidophilus (22.00 ± 0.816) total counts was closest to the control chlorhexidine (S. mutans =22.00 ± 0.876 and L. acidophilus =21.10 ± 0.876). A greater activity against S. mutans and L. acidophilus is seen in the zone of inhibition of the 60 µg/ml experimental concentration of bamboo shoot ethanolic extract, with a significant difference in the disc diffusion assay. Conclusion The treatment with bamboo shoot extract was equivalent effective in the mentioned bacterial species. Clinical relevance It can be assured that preventive measures like mouth rinse and dentifrices compromising bamboo shoots, a potential dental biomaterial, would be optimistic agents for caries control, including the cariostatic effect.

PREPARATION OF HEALTH BENEFICIAL PROBIOTIC SOYA ICE -CREAM AND EVALUATION OF QUALITY ATTRIBUTES

Probiotic soya ice cream was prepared by using food grade bacteria Lactobacillus paracasei S1(MF443873) and Lactobacillus rhamnosus KC6 (MF443875). Bacteria have been encapsulated in milk and sodium alginate beads to improve their viability in soya ice -cream. Viable cell number of entrapped probiotic isolates was observed with calcium chloride concentration of 0.1M at pH 7.0, hardening time of 8h. Morphological characterization of microencapsulated beads was done by using SEM. Probiotic soy ice-cream was stored at -20oC for 30 days and was investigated for sensory qualities, microbial count and physicochemical characteristics. Microencapsulated probiotic ice-cream was found more stable during storage as compared to those containing free probiotic microorganisms. As far as physico-chemical characteristics of probiotic enriched soy ice-cream is concerned, though changes in quality attributes viz. pH, ascorbic acid, lactic acid, total soluble solid and antioxidant content during storage interval have been observed but these changes were statistically non- significant.

Melitoxin Inhibits Proliferation, Metastasis, and Invasion of Glioma U251 Cells by Down-regulating F2RL1.

As the principal active component of bee venom, melittin has an anti-cancer effect in different cancers. This study was aimed to investigate the effect of melittin in glioma and explore whether F2RL1 is closely involved in glioblastoma cells proliferation. TCGA and GES databases were used to evaluate the role of F2RL1 in gliomas. The U251 cells were divided into a control lentivirus + PBS group (NC-PBS), F2RL1 intervention lentivirus + PBS group (KD-PBS), control lentivirus + melittin group (NC-melittin), and F2RL1 intervention lentivirus + melittin group (KD-melittin). Cell proliferation was detected by MTT and EDU staining assays. The apoptosis rate was assessed by flow cytometry. Expressions of genes related to apoptosis, cycle arrest, migration, and invasion were detected by qRT-PCR. Cellular LDH concentrations were detected by ELISA. The subcutaneous tumor volume of nude mice was analyzed by xenograft method. F2RL1 was significantly overexpressed in glioma tissues and were reduced in the melittin-treated group compared to the blank group. F2RL1 knockdown and melittin alone or in combination increased the proportion of cells in the G1-phase, and the combination was more pronounced. The KD-melittin group showed a decrease in the number of viable cells at 24, 48, 72, and 96 h compared to the NC-PBS group. The number of cell migration and invasion was decreased in the KD-melittin group compared to the other groups. Moreover, the genes related to cell cycle arrest and apoptosis were significantly changed in the KD-melittin group. At weeks 4, 5, and 6, the tumor volume in the KD-melittin group was smaller than that in the KD-PBS group and NC-melittin group. Interference with the target gene F2RL1 inhibited the proliferation of glioma U251 cells, and melittin treatment inhibited the proliferation of glioma U251 cells. Melittin inhibited the proliferation of glioma U251 cells by suppressing the expression of target gene F2RL1.

Hepatotoxicity evaluations of different surface charged carbon quantum dots in vivo and in vitro

Recently, carbon quantum dots (CQDs) have become popular because of their simple synthesis and potential applications. Although CQDs have high biocompatibility, their biotoxicity must be verified to reduce the possible risks associated with large-scale application. In this study, the hepatotoxicity of three CQD types, namely diammonium citrate (AC)-based (CQDs-AC), spermidine trihydrochloride (Spd)-based (CQDs-Spd), and AC- and Spd-based CQDs (CQDs-AC/Spd), were evaluated in vivo and in vitro. It was observed in vivo that CQDs-Spd and CQDs-AC/Spd, but not CQDs-AC, caused histopathological damage, including liver steatosis and mild mixed inflammatory cell infiltration; however, reduced liver function was only observed in CQD-Spd–treated mice. The in vitro results revealed that only CQDs-Spd significantly decreased the number of viable HepG2 cells (NADH depletion) and induced oxidative stress (heme oxygenase-1 activation) after 24 h of exposure, which promoted inflammatory factor secretion (NF-κB activation). Additionally, decreasing zonula occludens-2 and α1-antitrypsin protein expression in HepG2 cells suggested that CQD-Spd exposure increases the risk of liver diseases. Our results revealed that CQDs-Spd had greater hepatotoxic potential than CQDs-AC and CQDs-AC/Spd, which might be attributable to their high positive surface charge. Overall, the risk of CQD-induced hepatotoxic risk must be considered when applying positively charged CQDs.

Identification and quantification of viable Lacticaseibacillus rhamnosus in probiotics using validated PMA-qPCR method.

The identification and quantification of viable bacteria at the species/strain level in compound probiotic products is challenging now. Molecular biology methods, e.g., propidium monoazide (PMA) combination with qPCR, have gained prominence for targeted viable cell counts. This study endeavors to establish a robust PMA-qPCR method for viable Lacticaseibacillus rhamnosus detection and systematically validated key metrics encompassing relative trueness, accuracy, limit of quantification, linear, and range. The inclusivity and exclusivity notably underscored high specificity of the primers for L. rhamnosus, which allowed accurate identification of the target bacteria. Furthermore, the conditions employed for PMA treatment were fully verified by 24 different L. rhamnosus including type strain, commercial strains, etc., confirming its effective discrimination between live and dead bacteria. A standard curve constructed by type strain could apply to commercial strains to convert qPCR Cq values to viable cell numbers. The established PMA-qPCR method was applied to 46 samples including pure cultures, probiotics as food ingredients, and compound probiotic products. Noteworthy is the congruity observed between measured and theoretical values within a 95% confidence interval of the upper and lower limits of agreement, demonstrating the relative trueness of this method. Moreover, accurate results were obtained when viable L. rhamnosus ranging from 103 to 108 CFU/mL. The comprehensive appraisal of PMA-qPCR performances provides potential industrial applications of this new technology in quality control and supervision of probiotic products.

Viable tendon neotissue from adult adipose-derived multipotent stromal cells.

Background: Tendon healing is frequently prolonged, unpredictable, and results in poor tissue quality. Neotissue formed by adult multipotent stromal cells has the potential to guide healthy tendon tissue formation. Objectives: The objective of this study was to characterize tendon neotissue generated by equine adult adipose-derived multipotent stromal cells (ASCs) on collagen type I (COLI) templates under 10% strain in a novel bioreactor. The tested hypothesis was that ASCs assume a tendon progenitor cell-like morphology, express tendon-related genes, and produce more organized extracellular matrix (ECM) in tenogenic versus stromal medium with perfusion and centrifugal fluid motion. Methods: Equine ASCs on COLI sponge cylinders were cultured in stromal or tenogenic medium within bioreactors during combined perfusion and centrifugal fluid motion for 7, 14, or 21days under 10% strain. Viable cell distribution and number, tendon-related gene expression, and micro- and ultra-structure were evaluated with calcein-AM/EthD-1 staining, resazurin reduction, RT-PCR, and light, transmission, and scanning electron microscopy. Fibromodulin was localized with immunohistochemistry. Cell number and gene expression were compared between culture media and among culture periods (p < 0.05). Results: Viable cells were distributed throughout constructs for up to 21days of culture, and cell numbers were higher in tenogenic medium. Individual cells had a round or rhomboid shape with scant ECM in stromal medium in contrast to clusters of parallel, elongated cells surrounded by highly organized ECM in tenogenic medium after 21days of culture. Transcription factor, extracellular matrix, and mature tendon gene expression profiles confirmed ASC differentiation to a tendon progenitor-like cell in tenogenic medium. Construct micro- and ultra-structure were consistent with tendon neotissue and fibromodulin was present in the ECM after culture in tenogenic medium. Conclusion: Long-term culture in custom bioreactors with combined perfusion and centrifugal tenogenic medium circulation supports differentiation of equine adult ASCs into tendon progenitor-like cells capable of neotissue formation.

Zinc phthalocyanine loaded- antibody functionalized nanoparticles enhance photodynamic therapy in monolayer (2-D) and multicellular tumour spheroid (3-D) cell cultures.

In conventional photodynamic therapy (PDT), effective delivery of photosensitizers (PS) to cancer cells can be challenging, prompting the exploration of active targeting as a promising strategy to enhance PS delivery. Typically, two-dimensional (2-D) monolayer cell culture models are used for investigating targeted photodynamic therapy. However, despite their ease of use, these cell culture models come with certain limitations due to their structural simplicity when compared to three-dimensional (3-D) cell culture models such as multicellular tumour spheroids (MCTSs). In this study, we prepared gold nanoparticles (AuNPs) that were functionalized with antibodies and loaded with tetra sulphonated zinc phthalocyanine (ZnPcS4). Characterization techniques including transmission electron microscopy (TEM) was used to determine the size and morphology of the prepared nanoconjugates. We also conducted a comparative investigation to assess the photodynamic effects of ZnPcS4 alone and/or conjugated onto the bioactively functionalized nanodelivery system in colorectal Caco-2 cells cultured in both in vitro 2-D monolayers and 3-D MCTSs. TEM micrographs revealed small, well distributed, and spherical shaped nanoparticles. Our results demonstrated that biofunctionalized nanoparticle mediated PDT significantly inhibited cell proliferation and induced apoptosis in Caco-2 cancer monolayers and, to a lesser extent, in Caco-2 MCTSs. Live/dead assays further elucidated the impact of actively targeted nanoparticle-photosensitizer nanoconstruct, revealing enhanced cytotoxicity in 2-D cultures, with a notable increase in dead cells post-PDT. In 3-D spheroids, however, while the presence of targeted nanoparticle-photosensitizer system facilitated improved therapeutic outcomes, the live/dead results showed a higher number of viable cells after PDT treatment compared to their 2-D monolayer counterparts suggesting that MCTSs showed more resistance to PS drug as compared to 2-D monolayers. These findings suggest a high therapeutic potential of the multifunctional nanoparticle as a targeted photosensitizer delivery system in PDT of colorectal cancer. Furthermore, the choice of cell culture model influenced the response of cancer cells to PDT treatment, highlighting the feasibility of using MCTSs for targeted PS delivery to colorectal cancer cells.

Cereal-based fermented synbiotic instant powders: a dessert practice

Objective: This study was aimed to develop synbiotic and probiotic instant powder mixtures that can be used in food products to meet the increasing interest of consumers in functional foods. Materials and Methods Cereal flours (oat, rice, wheat), sesame, cranberry, chestnut and milled germinated grains (lentil, mung bean) were blended and autoclaved then subjected to fermentation process using the Lactobacillus plantarum strain. A part of the porridge was conventionally dried (FPP) and skim milk powder+fructooligosaccharide+inulin mixture as prebiotic sources were added to the other part and subjected to freeze drying (FSP). The viability of probiotics in the gastrointestinal environment and the Angiotensin-converting enzyme inhibition and bile-acid binding capacities of the powders after in vitro digestion were analyzed. The sensory acceptability of the desserts was evaluated. Results: FSP contained a higher number of viable cells than FPP after in vitro digestion. Relative bile-acid binding and angiotensin-converting enzyme inhibition capacities of samples were confirmed as their cholesterol-lowering and blood pressure-regulating potential. The panelists rated the dessert samples enriched with 5-15% FPP and FSP as 'liked'. Conclusion: The integration of these powders into a variety of food products will provide consumers with healthier dietary choices that support their overall health goals.

Evaluation of pan‐epithelial viability in the whole porcine lens

Purpose: The lens has a monolayer of epithelial cells on its anterior surface that contributes to its homeostasis. A quantification of the viability of these cells is essential to evaluate the safety of new therapies targeting the lens (cataract, presbyopia). The observation of this whole tissue is made difficult by its optical properties (transparency and refraction) and only simplified models have been used until now to measure epithelial viability (primary cell cultures or partial surgical specimens).Aims: To develop a method to quantify the viability of epithelial cells on whole ex vivo lenses.Methods: Ten fresh pairs of 6‐month‐old porcine lens were retrieved from local slaughterhouse. For each pair, one lens was randomly selected to undergo a localized lesion and the other lens remained intact. The lesion was made on the anterior surface with a steel hexagonal rod (to make the mark easily identifiable) previously immersed in liquid nitrogen for one minute. Both lenses were incubated for 1 h at 20°C in a HEC mixture. Images were acquired with a macroscope and analysed with ImageJ. Calcein‐AM and Ethidium images were used to calculate the area covered by living epithelial cells. Hoescht images allowed to count cell nuclei per unit area. Viable epithelial cell density (vECD) was defined as the number of viable cells per unit area. The lenses were immersed (isotonic saline) inside a black container during the measurements to minimize reflections and refractive phenomena or drying of the samples.Results: The vECD median on the ten pairs was 2840 cells/mm2 [10th‐90th percentiles = 2479–3494] for cryo‐applied lenses versus 3364 cells/mm2 [2919–3739] for healthy lenses (p = 0.002).Conclusions: The triple HEC staining adapted to the whole lens provides an unrivalled measure of pan‐epithelial viability without the need for a tedious capsule dissection step that destroys the lens architecture. Other studies will be necessary to determine its ability to discriminate small variations or type of mortality.

Exploration of Klebsiella aerogenes derived secondary metabolites and their antibacterial activities against multidrug-resistant bacteria.

As the effectiveness of current treatments against the development of antimicrobial resistance is declining, new strategies are required. A great source of novel secondary metabolites with therapeutics effects are the endophytic bacteria present in medicinal plants. In this study, Klebsiella aerogenes (an endophytic bacteria belonging to the Enterobacteriaceae family) was isolated from Kalanchoe blossfeldiana (a medicinal plant". The bacterial secondary metabolites were identified using GC-MS techniques. Furthermore, the antibacterial potentials were investigated against multi-drug resistance (MDR) Salmonella typhi and Staphylococcus aureus. The GC-MS chromatogram of K. aerogenes secondary metabolites extract displayed total of 36 compounds. Ethyl acetate extracts of K. aerogenes, showed mean zone of growth inhibition of 15.00 ± 1.00 against S. typhi and 7.00 ± 1.00mm against S. aureus, respectively. The extract demonstrated significant antibacterial effectiveness against S. typhi and moderate antibacterial efficacy against S. aureus, with minimum inhibitory concentration (MIC) values ranging from 0.089 to 0.39 mg/mL. The time-kill kinetics profile of the ethyl acetate extract against S. typhi revealed a decrease in the number of viable cells during the initial 5, 6, and 24 hours. Conversely, there was a sudden increase in viable cells up to 6 hours for S. aureus. The identified secondary metabolite with high percentage than others, benzeneethanamine exhibited favorable interactions (-7.2 kcal/mol) with the penicillin-binding protein (PBP2a) of S. aureus and (-7.5 kcal/mol) osmoporin (OmpC) of S. typhi, indicating its potential as a candidate for drug development against these MDR bacteria. This study reported for the first time, bacterial endophytes associated with K. blossfeldiana with antibacterial activities.

Preparation of a Lactobacillus rhamnosus ATCC 7469 microencapsulated-lactulose synbiotic and its effect on equol production.

Equol is a highly active product of soy isoflavones produced by specific bacteria in the human or animal colon. However, equol production is influenced by differences in the gut flora carried by the body. Our previous research has shown that a synbiotic preparation comprising the probiotic Lactobacillus rhamnosus ATCC 7469 and the prebiotic lactulose can enhance equol production by modulating the intestinal flora. Nevertheless, the harsh environment of the gastrointestinal tract limits this capability by diminishing the number of probiotics reaching the colon. Microencapsulation of probiotics is an effective strategy to enhance their viability. In this study, probiotic gel microspheres (SA-S-CS) were prepared using an extrusion method, with sodium alginate (SA) and chitosan (CS) serving as the encapsulating materials. Scanning electron microscopy (SEM) was employed to observe the surface morphology and the internal distribution of bacteria within the microcapsules. The structural characteristics of the microcapsules were investigated using Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Furthermore, the thermal stability, storage stability, probiotic viability post-simulated gastrointestinal fluid treatment, and colon release rate were examined. Finally, the impact of probiotic microencapsulation on promoting equol production by the synbiotic preparation was assessed. The results indicated that the microcapsules exhibited a spherical structure with bacteria evenly distributed on the inner surface. Studies on thermal and storage stability showed that the number of viable cells in the probiotic microcapsule group significantly increased compared to the free probiotic group. Gastrointestinal tolerance studies revealed that after in vitro simulated gastrointestinal digestion, the amount of viable cells in the microcapsules was 7 log10 CFU g-1, demonstrating good gastrointestinal tolerance. Moreover, after incubation in simulated colonic fluid for 150 min, the release rate of probiotics reached 93.13%. This suggests that chitosan-coated sodium alginate microcapsules can shield Lactobacillus rhamnosus ATCC 7469 from the gastrointestinal environment, offering a novel model for synbiotic preparation to enhance equol production.

Влияние ионизирующего излучения на активность антиоксидантных ферментов штамма Escherichia coli «ПЛ-6» при многократном облучении

The article presents the analysis of enzymatic activity of Escherichia coli “PL-6” cells following multiple gamma irradiation with increasing doses. For irradiation the stationary gamma ray unit “Researcher” (Co-60) with the dose rate of 36.7 Gy/min was used. Radiation doses to the bacterial cells increased from 3.5 to 10 kGy in 0.5 kGy increment, and from 10 to 30 kGy in 5 kGy increment. Unirradiated suspended bacterial cells were used as control. The irradiated cells enzymatic activity was estimated with the spectrophotometric method on the "MultiSkan Go" device (Thermo Fisher Scientific, Finland). The results of the study demonstrate the increase in the E. coli ”PL-6” cells radioresistance caused by gamma irradiation of the cells with gradually increased doses. Exposure of the bacteria to a higher gamma radiation dose caused decrease in the number of viable cells from 10Е+9 CFU/ml in the control culture sample to 10Е+2 CFU/ml after irradiation at a dose of 30 kGy. The most radiation-resistant cells were selected from cells undergone successive multiple irradiation with constantly increasing dose of ionizing radiation. The enzymes of the antioxidant system in irradiated cells exhibit increased specific activity as compared to the enzymes in control, unirradiated, cells. The activity of enzymes increases with an increase in the radiation dose. Catalase activity increased after multiple irradiation by 5 times and peroxidase activity increased by 4 times. To compare the results in the groups, the Student's t-test was applied at p&lt;0.05.

Combination of bifidobacteria and arabinogalactan when fermenting skim milk

The work investigated the fermentation of skim milk with a concentrate of bifidobacteria (Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium breve and Bifidobacterium infantis) in the presence of arabinogalactan in amounts of 3, 5, 7 and 10%. It was found that after fermentation at a temperature of 37 °C for 12 hours, the characteristic sour-milk taste and curd were not observed in any of the samples, regardless of the presence of arabinogalactan. It was revealed that after storage at a temperature of (4±2) °C for 10 days, the consistency of the control and test samples remained homogeneous, without separation and visible syneresis. The increase in titratable acidity at the post-oxidation stage for 10 days was no more than 1-2 °T. It was shown that, under experimental conditions, the number of viable bifidobacteria cells was greater in test samples containing from 5 to 10% arabinogalactan than in control variants and in samples with a mass fraction of arabinogalactan of 3%. Based on the totality of the observed results, it can be assumed that the prebiotic was used by microorganisms as a source of nutrition.

Volatilization and disinfection efficacy of gaseous hypochlorous acid from an air washer-type humidifier in a large space.

An air washer-type humidifier has two useful functions: humidification, and air purification, and it applies to large indoor spaces. In this study, the efficacy of an air washer-type humidifier fed with 24 L of weakly acidic electrolyzed water（WAEW) at pH 5.0 and 30 mg/L in disinfecting attached bacteria and airborne microorganisms was studied in a 480 m3 indoor space. The humidifier was operated at a shower volume of 9.0 L/min of WAEW and at an air flow rate of 29 m3/min. Volatilization of gaseous hypochlorous acid（HOCl(g)) proceeded according to first-order kinetics during the 60 min of operation. Fresh WAEW was supplied to the humidifier every 60 min, and the HOCl(g) concentration in the indoor space was maintained within the range of 25-52 ppb for at least 180 min of operation. The number of viable bacterial cells on wet agar plates placed on the floor at a distance of 5-20 m away from the humidifier decreased by 2.0-3.0 log after 30 min of operation, and no viable cells were detected after 60 min of operation. A logarithmic reduction of more than 2.7 was achieved within 15 min against bacteria-attached plates placed at a 1.5 m-height position where the outlet airflow from the humidifier was directly exposed. This indicates that the disinfection efficacy of HOCl(g) volatilized from the humidifier depends on the rate of outlet airflow reaching the bacteria-attached plates. The number of viable airborne microorganisms decreased by approximately 54% after 180 min of operation. This study demonstrated that an air-washer-type humidifier can spread HOCl(g) evenly throughout a large indoor space and is effective in disinfecting attached bacteria and airborne microorganisms.

Vikil 20 Suppresses the Proliferation of Prostate Cancer (PC-3) Cells and Quenches Free Radicals In Vitro.

Vikil 20 is an herbal formula produced in Ghana and is widely marketed as a product to boost immunity as well as for general well-being. However, the pharmacological effect of this herbal preparation has not been proven scientifically. Therefore, this study was aimed at investigating the antioxidative as well as the anti-prostate cancer effects of the product. To assess the antioxidative effect of Vikil 20, the DPPH and ABTS activities were investigated. The total phenolic content was investigated using the Folin-Ciocalteu method. The cytotoxic effect of Vikil 20 against prostate cancer (PC-3) cells as well as normal (RAW 264.7) cells was investigated using the MTT assay whereas its anti-metastatic effect was analyzed using the cell migration assay. The effect of Vikil 20 on cell adhesion was analyzed via the cell adhesion assay whereas its effect on TNF-α secretion was investigated using a TNF-α detection kit. Vikil 20 demonstrated significant antioxidant effects by suppressing 57.61% and 92.88% respectively of DPPH and ABTS radicals at 1000 µg/mL with total phenolic contents of 140.45 mg GAE/g. Vikil 20 suppressed the proliferation of PC-3 cells by reducing the number of viable cells to 49.5% while sparing the RAW, 264.7 cells. Further, Vikil 20 significantly suppressed both cellular migration and adhesion of prostate cancer cells. Finally, suppression of cellular migration and adhesion is associated with a reduction in TNF-α secretion by PC-3 cells. Taken together, Vikil 20 was found to possess significant antioxidant and anti-prostate cancer effects in vitro.

Cytotoxicity of Sambucus nigra L. on Cancer Cell Line and In Vitro Antioxidant Properties

Objective: Essential oils, free fatty acids, flavonoids, glycosides, phenolic acids, carotenoids, vitamins, and minerals are found in significant quantities in the characteristic chemical composition of Sambucus nigra L. This study aimed to evaluate the antioxidant potential of Sambucus nigra L. Fructus and evaluate the cytotoxicity on the cancer cell line.&#x0D; Methods: The Sambucus nigra L. fruits were collected from Yalova Atatürk Horticultural Central Research Institute in September 2021. The ethanol extract was prepared. Antioxidant property of Sambucus nigra L. fruit extracts was evaluated with 2,2-diphenyl-1-picryl-hydrazyl- hydrate (DPPH) radical scavenging activity, Cupric Ion Reducing Antioxidant Capacity (CUPRAC). Also, total phenolic content, total flavonoid content, and total anthocyanin content were calculated. Liver hepatocellular carcinoma cell line (HepG2) was used for cytotoxicity assay and an 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was applied.&#x0D; Results: The total phenolic, total flavonoid contents and total monomeric anthocyanins were 9.75±0.92 mg GAE/ mg fruit extract, 0.07437±0.004 mg quercetin/ mg fruit extract, respectively. 0.45±0.0014 mg catechin/gram of fruit extract and 2.08±0.025 mg Cyanidin-3 glucosides/g fruit extract. CUPRAC and DPPH results showed that Sambucus nigra L. extract has strong antioxidant activity. The results of the cytotoxicity assay indicated that while concentrations of the extract increased, the viability of HepG2 decreased.&#x0D; Conclusion: Our findings suggest that the Sambucus fruit extract is particularly rich in antioxidant components that are possibly modulating their beneficial use for hepatocellular malignancies, significantly reducing the number of viable cancer cells and inducing cell death.

Prebiotic Cellulose-Pullulan Matrix as a "Vehicle" for Probiotic Biofilm Delivery to the Host Large Intestine.

This study describes the development of a new combined polysaccharide-matrix-based technology for the immobilization of Lactobacillus rhamnosus GG (LGG) bacteria in biofilm form. The new composition allows for delivering the bacteria to the digestive tract in a manner that improves their robustness compared with planktonic cells and released biofilm cells. Granules consisting of a polysaccharide matrix with probiotic biofilms (PMPB) with high cell density (>9 log CFU/g) were obtained by immobilization in the optimized nutrient medium. Successful probiotic loading was confirmed by fluorescence microscopy and scanning electron microscopy. The developed prebiotic polysaccharide matrix significantly enhanced LGG viability under acidic (pH 2.0) and bile salt (0.3%) stress conditions. Enzymatic extract of feces, mimicking colon fluid in terms of cellulase activity, was used to evaluate the intestinal release of probiotics. PMPB granules showed the ability to gradually release a large number of viable LGG cells in the model colon fluid. In vivo, the oral administration of PMPB granules in rats resulted in the successful release of probiotics in the colon environment. The biofilm-forming incubation method of immobilization on a complex polysaccharide matrix tested in this study has shown high efficacy and promising potential for the development of innovative biotechnologies.

Memantine protects the cultured rat hippocampal neurons treated by NMDA and amyloid β1-42.

Alzheimer's disease (AD) is a devastating neurodegenerative condition with no effective treatments. Recent research highlights the role of NMDA receptors in AD development, as excessive activation of these receptors triggers excitotoxicity. Memantine, an NMDA receptor antagonist, shows promise in curbing excitotoxicity. What sets our study apart is our novel exploration of memantine's potential to protect hippocampal neurons from neurotoxicity induced by NMDA and amyloid β1-42, a hallmark of AD. To achieve this, we conducted a series of experiments using rat hippocampal cell cultures. We employed Hoechst and propidium iodide double staining to assess neuronal viability. Analyzing the viability of neurons in normal conditions compared to their status after 24 h of exposure to the respective agents revealed compelling results. The incubation of hippocampal neurons with NMDA or amyloid β1-42 led to a more than twofold increase in the number of apoptotic and necrotic neurons. However, when memantine was co-administered with NMDA or amyloid β1-42, we witnessed a notable augmentation in the number of viable cells. This unique approach not only suggests that memantine may act as a neuroprotective agent but also emphasizes the relevance of hippocampal neuron cultures as valuable models for investigating excitotoxicity and potential AD treatments.