Percentage Of Surviving Cells Research Articles

Abstract 1559: Repurposing of pimavanserin tartrate for the treatment of glioblastoma

Abstract Glioblastoma is a highly aggressive brain tumor that has a median survival of 15 months. We aim to give a substitute to the current chemotherapeutic drug temozolomide by using the FDA-approved anti-psychotic pimavanserin tartrate (PVT). To uncover the effectiveness of PVT, we did cytotoxicity assays using SRB to determine the percentage of cell survival in a dose-dependent and time-dependent manner. The apoptotic effects were measured using Annexin-V/APC assay. The mechanism of action was determined utilizing Western Blotting. In different glioblastoma tumor cell lines, the IC50 for 24 hours was between 6 µM and 10 µM. 48 and 72 hours had an IC50 between 4 µM and 6 µM. SF188 glioblastoma cell line had approximately 70% to 80% more apoptotic cells in 10 µM PVT when compared to control in Annexin-V/APC assay. PVT causes a strong modulation of the PI3K/AKT and MAPK pathways when given in increasing concentration at 48 hours. The level of apoptosis was confirmed through Western Blot as well using apoptotic markers. The preliminary data show the ability of PVT to cause apoptosis of glioblastoma tumor cells in vitro through the AKT and MAPK signaling. Citation Format: Carson Zabel, Sharavan Ramachandran, Sanjay Srivastava. Repurposing of pimavanserin tartrate for the treatment of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1559.

Fabrication of neuroprotective silk-sericin hydrogel: potential neuronal carrier for the treatment and care of ischemic stroke

Ischemic stroke results in severe disabilities due to extensive cellular loss and the resulting impairment of brain functions. Current methods for regenerating brain tissue are ineffective. Stroke treatment requires innovative therapeutic techniques that are both safe and effective. For neuronal repair, a promising alternative is using a hydrogel-based tissue engineering technique that delivers neurotrophic cytokines and cells to injured sites. However, the limited encapsulation effectiveness, less in vivo cell survival ratio and cytokine loss make this strategy difficult to implement. We aim to design a biomaterial that can efficiently construct a matrix enriching the survival of cells and minimizing loss in vivo cytokines to overcome these constraints. We report the development of genipin conjugated sericin hydrogels (Gen-SH) with a high porous morphology and a moderate swelling rate utilizing sericin, a natural silk protein. In vitro, Gen-SH aids in the attachment and development of neurons. Our results indicate that sericin is inherently neuroprotective and neurotrophic, branching and publicizing axon extension and avoiding hypoxia-induced cell death in primary neurons. Notably, the breakdown products of Gen-SH inherit these capabilities, saving the expense of cytokines. Furthermore, we show that the Lkb1–Nuak1 pathway is required for this neurotrophic impact, whereas the Bcl-2/Bax protein ratio is necessary for the neuroprotective effect. Transplanted in vivo, Gen-SH has a high percentage of cell survival and promotes cell proliferation. Taking all this information into account, it's clear that Gen-SH can serve as a viable carrier for treatment and care for ischemic stroke healing, both in terms of delivering neuronal cells and protecting them from oxidative damage.

Antibacterial activity of plant essential oils against indigenously characterized methicillin-resistant Staphylococcus aureus (MRSA).

Plant essential oils were evaluated for antimicrobial activity against Methicillin-resistant Staphylococcus aureus (MRSA). The isolates (n=03) were procured from Institute of Microbiology, UVAS Lahore, Pakistan. After biochemical and 16S rRNA gene-based PCR characterization, accession numbers were retrieved from NCBI i.e. MW344063.1, MW344064.1 and MW344065.1. These isolates exhibited molecular positivity by multiplex PCR for mecA, coa and eta toxin genes. Moreover, these isolates exhibited resistance to cefoxitin, ampicillin, amoxicillin, penicillin, amoxicillin clavulanate, ciprofloxacin, erythromycin and gentamicin. The antibiotic resistant isolates were evaluated for antimicrobial activity of plant essential oils. The highest zone of inhibition (mean ZOI±S.D.) was measured for Cinnamomum verum (22.67±1.52 mm) followed by Eucalyptus globulus (18.67±2.51 mm) and Syzygium aromaticum (12.67±2.51 mm). Lowest mean MIC value (0.33±0.11 mg/mL) was recorded for E. globulus . Eucalyptus globulus was processed for fractionation by column chromatography and n-hexane, chloroform, n-hexane + chloroform and ethyl-acetate fractions were evaluated for antibacterial activity. Lowest mean MIC (10.04±5.80 mg/mL) was recorded for E. globulus n-hexane fraction. Cell survival percentage of BHK21 cell line was 51.7% at 54.87mg/mL concentration of E. globulus n-hexane fraction. Through gas chromatography-mass spectrometry (GC-MS) analysis of n-hexane fraction, benzene was found abundant (29.9%) as active compound. It was concluded that E. globulus n-hexane fraction exhibited significantly promising results against MRSA .

Antibacterial and cytotoxic evaluation of sequential extract of Moringa oleifera leaves

Moringa Oleifera is an interesting plant used in Asian traditional medicine. In this study, in vitro antibacterial and cytotoxic evaluation of sequential extracts (aqueous, ethanol, and chloroform) of Moringa Oleifera was carried out. The antibacterial analysis was estimated with minimum inhibitory concentration (MIC) by micro dilution of Moringa Oleifera against common poultry pathogens Clostridium perfringens type A and Escherichia coli, while cytotoxic evaluation was estimated by the reduction in cell viability due to apoptosis or necrosis by metabolic events in the presence and absence of crude extracts or tested component. The aqueous extract shows the highest percentage yield (45/50gm) succeeded by ethanol extract (5.5gm/50gm) and chloroform extract (0.2gm/50gm). In our study, the zone of inhibition of sequential extracts of Moringa Oleifera against Haemophilus species are highest for chloroform (17mm), intermediate for ethanol (13mm), and lowest for aqueous extract (12.3mm). For chloroform extract the CSP was calculated at 10 different concentrations, 2000 µg/ml, 1000µg/ml, 500µg/ml, 250µg/ml, 125µg/ml, 62.5µg/ml, 31.25µg/ml, 15.63µg/ml, 7.81µg/ml and 3.91µg/ml .The results for cell survival percentage (CSP) in the present research are 26%, 46%, 58%, 55%, 60%, 63%, 62%, 59%, 68% and 82% respectively. The CSP results of chloroform extract indicated that it is toxic for cells at ≥1000µg/ml. At 1000µg/ml concentration CSP was 46% which is > 50% and therefore it is cytotoxic. At higher concentrations, chloroform is more cytotoxic than hexane because at > 1000µg/ml the cell survival percentage was recorded to be < 50%. For ethanol extract CSP was calculated at 10 concentrations, 6000µg/ml, 3000µg/ml, 1500µg/ml, 750µg/ml, 375µg/ml, 187.5µg/ml, 93.75µg/ml, 46.85µg/ml, 23.43µg/ml and 11.71µg/ml. The CSP values are 18%, 48%, 60%, 58%, 69%, 56%, 59%, 74%, 57% and 78% respectively which indicate that at concentrations ≥3000µg/ml the chloroform extract is toxic for cells. At a concentration less than 3000µg/ml, the CSP is more than 50%. So, as compared to hexane and chloroform, ethanol extract is less toxic at higher concentrations. Cytotoxicity of aqueous extract was calculated at 10 concentrations, 5000µg/ml, 2500µg/ml, 1250µg/ml, 625µg/ml, 312.5µg/ml, 156.25µg/ml, 78.125µg/ml, 39.06µg/ml, 19.53µg/ml and 9.76µg/ml. The CSP values are 8%, 18%, 42%, 56%, 54%, 59%, 55%, 62% 59% and 66% respectively. At a concentration ≥625µg/ml, the aqueous extract is toxic for cells. The CSP at 625µg/ml is 42%, hence toxic for cells. The cell survival percentage is more than 50% at a concentration > 625µg/ml, indicating that aqueous extract is more toxic to the cell than the rest of the three (Hexane, Chloroform, Ethanol extracts) at higher concentrations.

In Vitro Investigation of the Antioxidant and Cytotoxic Potential of Tabernaemontana ventricosa Hochst. ex A. DC. Leaf, Stem, and Latex Extracts

Tabernaemontana ventricosa (Apocynaceae) a latex-bearing plant is used in traditional medicine for its therapeutic benefits in reducing fever and hypertension and wound healing. Due to limited information on the plant’s pharmacological activities, this study aimed to investigate the antioxidant potential of the leaf, stem, and latex extracts of T. ventricosa, using the Folin-Ciocalteu (total phenolics), aluminum chloride colorimetric (total flavonoids), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The cytotoxic activity was evaluated in the human HEK293 (embryonic kidney), HeLa (cervical carcinoma), and MCF-7 (breast adenocarcinoma) cell lines using the MTT assay. The latex extracts possessed the highest total phenolic content (115.36 ± 2.89 mg GAE/g), followed by the stem hexane extracts (21.33 ± 0.42 mg GAE/g), the chloroform leaf (7.89 ± 0.87 mg GAE/g), and the chloroform stem (4.69 ± 0.21 mg GAE/g) extracts. The flavonoid content was substantially high ranging from 946.92 ± 6.29 mg QE/g in the stem hexane, 768.96 ± 5.43 mg QE/g in the latex, 693.24 ± 4.12 mg QE/g in the stem chloroform, and 662.20 ± 1.00 mg QE/g in the leaf hexane extracts. The DPPH assays showed the highest percentage of inhibition at 240 µg/mL, for the stem hexane (70.10%), stem methanol (65.24%), and stem chloroform (60.26%) extracts, with their respective IC50 values of 19.26 µg/mL (stem hexane), 6.19 µg/mL (stem methanol), and 22.56 µg/mL (stem chloroform). The FRAP assays displayed minimal inhibition ranging from 4.73% to 14.40%, except for the latex extracts which displayed moderate inhibition at 15 µg/mL (21.82%) and substantial inhibition at 240 µg/mL (98.48%). The HeLa and MCF-7 cell lines were the most sensitive to the extracts, with the hexane, chloroform, and methanol leaf and stem, and latex extracts significantly affecting the percentage cell survival. Overall, the various parts of T. ventricosa exhibited strong antioxidant activity correlating to its cytotoxicity. Further studies should focus on the isolation of specific antioxidant compounds that could be investigated for their anticancer potential.

Preparation and Preliminary Evaluation of Silver-Modified Anodic Alumina for Biomedical Applications

The present study reports a specific method for preparation of silver-modified anodic alumina substrates intended for biomaterial applications. Al2O3 coatings were obtained by anodization of technically pure aluminum alloy in sulfuric acid electrolyte. Silver deposition into the pores of the anodic structures was carried out employing in situ thermal reduction for different time periods. The obtained coatings were characterized using scanning electron microscopy (SEM), potentiodynamic scanning after 168 h in 3.5% NaCl solution and bioassays with human fibroblast and NIH/3T3 cell lines. The modified alumina substrates demonstrated better biocompatibility compared to the control anodic Al2O3 pads indicated by increased percent cell survival following in vitro culture with human and mouse fibroblasts. The Ag-deposition time did not affect considerably the biocompatibility of the investigated anodic layers. SEM analyses indicated that mouse NIH/3T3 cells and human fibroblasts adhere to the silver-coated alumina substrates retaining normal morphology and ability to form cell monolayer. Therefore, the present studies demonstrate that silver coating of anodic alumina substrates improves their biocompatibility and their eventual biomedical application.

Development of Innovative Gel Sunscreen Products from Momordica cochinchinensis (Lour.) Spreng. Extract

A study on the development of sunscreen gel products from Momordica cochinchinensis(Lour.) Spreng. extract aimed to study phenolic content, inhibitory effect of Elastase and Tyrosinase, product stability, toxicity, astringent effect of M. cochinchinensis extract, skin elasticity value, suitable product formula calculation for preparing sunscreen gel products from M. cochinchinensis extract, and irritation test. The process started fromthe selection of raw materials, preparation of extracts for determining the total phenolic content, development of suitable formula, test of safety and product physical characteristics, and then test of the anti-allergic effect of 10 volunteers to get efficient and safe sunscreen gel from M. cochinchinensis extract. The study result indicated that M. cochinchinensisaril extract had antioxidant activity DPPH of 1.51±0.05 mg/ml, compared to standard substance - Vitamin C, and total phenolic content of 13.18±0.18 (mg equivalent of gallic acid per 100 g - dry weight). Regarding Cytotoxicity at a concentration of 0.0001-1 mg/ml, it revealed that M. cochinchinensisaril extract was not toxic to human skin cells with the cell survival percentage at a concentration of 1 mg/ml equaled to 95.35±1.86 and 88.15±4.73%, respectively. M. cochinchinensisaril extract concentration of 1 mg/ml had astringent effect which can stimulate human skin cells to move together faster than the control group but showed effect slower than Vitamin C concentrate of 1 mg/ml. and did not have inhibitory effect on Elastase and Tyrosinaseenzymes. Regarding M. cochinchinensis seed oil extract, it did not toxic to human skin cells at the concentration of 0.0001-1 mg/ml with the survival percentage equaled to 105.67-111.46%, and had a few antioxidants activity of unsaturated fatty acids with an IPC50 more than 1000 mg/ml. This study was only the development of sunscreen gel products from M. cochinchinensis extract.

Protective Effect of Cinnamaldehyde on METH-induced Neurotoxicity in PC12 Cells via Inhibition of Apoptotic Response and Oxidative Stress.

Methamphetamine (METH) is a potent central nervous system (CNS) stimulant and frequently used illegal drugs. Repeated exposure to METH can induce degenerative changes in dopaminergic and serotonergic axons. There is no standard medical treatment for METH’s neurotoxic effects. Cinnamaldehyde is an important compound of cinnamon and has activities against neurological disorders. The present study was designed to examine the neuroprotective effect of trans-cinnamaldehyde (TCA) on METH-induced cytotoxicity. PC12 cells were treated with METH (2.5 mM) 24 h after treated with different concentrations of TCA (3.75- 50 μM). The percentage of cell survival was evaluated by MTT assay and the following parameters were measured to detect apoptosis and oxidative stress responses: DNA fragmentation, ROS production and GSH content. Exposure to 2.5 mM METH decreased the cell viability and GSH levels, caused the generation of reactive oxygen species and ultimately induced apoptosis. Pretreatment with TCA at 3.125-25 μM significantly attenuated cell viability loss. TCA, especially at a concentration of 12.5 and 25 μM, decreased the apoptosis and ROS generation and increased the GSH level compared with the METH group. The findings of the present study suggested that TCA exerted a protective effect against METH-induced neurotoxicity through mechanisms related to antioxidant and anti-apoptosis. It is suggested that TCA may be useful for the prevention and treatment of harmful effects of METH on the brain.

Phenolic Profile and Biological Properties of <i>Momordica charantia</i>

<i>M. charantia</i> is an important medicinal plant belongs to family <i>cucurbitaceae</i>. It originates from India, Malasiya and is widely spread all over tropical, subtropical and warm temperate regions of the world. This research work has been designed to evaluate the antioxidant, antimicrobial and toxicological potential of <i>M. charantia. </i>The antifungal and antioxidant components of<i> M. charantia</i> leaves, seeds and peels were extracted by using four solvent systems (80% methanol, 80% ethanol, 100% methanol and 100% ethanol) andleaves presented maximum extract yield (22.7 g/100g DW) in 80% methanolic solvent system. Phytochemical analysis of <i>M. Charantia </i>leaves, seeds and peels extracts performed in terms of total phenolic and total flavonoid contents, showed that 80% methanolic leaves extract offered highest total phenolic contents (47.1 mg GAE/g DW), whereas80% ethanolic leaves gave maximum total flavonoid contents (67.3 mg CE/g DW). The phenolic contents were also analysed by HPLC. Antioxidant activity was determined by DPPH radical scavenging activity and measure of reducing power. Results revealed that 80% methanolic leaves extract showed highest radical scavenging activity and reducing potential. Antimicrobial activity of <i>M. charantia </i>leaves, seeds and peels was investigated by Disc Diffusion Method and Minimum Inhibitory Concentration (MIC). Results showed that 80% methanolic extract of leaves exhibited highest antibacterial and antifungal potential against <i>P. multocida</i> (30 mm DIZ) and <i>A. paraciticus</i> (28 mm DIZ), respectively. Cytotoxicity analysis was performed on BHK-21 cell by adopting the MTT assay. The cytotoxicity activity of the 80% methanolic extract of leaves was evaluated by noticing the cell survival percentage (53.4%). Overall results of the present study showed that 80% methanolic leaves extracts of<i> M. charantia</i> possesses very good antioxidant, antimicrobial and cytotoxic properties.

Effect of TiO2 Thin Film Coating on AZ91D Alloy and Investigation of Corrosion Behavior, Mechanical Properties, and Biocompatibility

Today magnesium and its alloys have applications in different industries and in addition to this they are used as orthopedic implants. Some of the magnesium alloys can be used as biodegradable implants in the body. Coating of the ceramic material on the magnesium alloys is one of the methods to increase the lifetime of these alloys. In this research, TiO2 thin film was coated on the surface of the AZ91D using the sputtering method. Morphology and cross-section of the coating were done using Field emission scanning electron microscopy (FESEM), X-ray diffraction analysis (XRD), Vickers hardness test (VM), roughness testing (RT) to investigate the properties of corrosion, polarization tests of corrosion in a simulated body fluid (SBF), adaptability of the sample in the body environment from the toxicity assessment test (MTT) and cellular adhesion. Results from the corrosion test show that the corrosion rate of the coated samples with TiO2 is improved compared to the samples without coating. The percentage of cell survival on the surface of the samples was increased from 80% for the uncoated sample to 90% for the coated sample with TiO2, and the hardness of the coated sample was 119 Hv, and its roughness was 0.12 µm. Applying the coating by increasing the hardness and real contact area and reducing the friction coefficient can increase the wear resistance of the substrate.

The role of mTOR and oral intervention of combined Zingiber officinale-Terminalia chebula extract in type 2 diabetes rat models.

The present study examined the potential of Zingiber officinale-Terminalia chebula extract alone (ZO and TC) and in combination (ZOTC) against type 2 diabetes via downregulation of mechanistic target of rapamycin (mTOR). The 1:4 (ZOTC) ratio showed high cell survival percentage against the rat insulinoma cell line (RIN-5F) when compared to other possible ratios of ZOTC. Oral administration of ZO alone, TC alone, combined ZOTC (1:4), and the positive control metformin (Met) in fructose-streptozotocin (STZ) -induced diabetic rats showed reduced blood glucose levels, reduced insulin resistance (HOMA-IR), increased insulin levels, and increased pancreatic beta cell function (HOMA-β). ZOTC treatment in diabetic rats ameliorated the antioxidant status without affecting liver and serum parameters. Histological evaluation of the pancreas was performed to find pathological changes; the transcriptional and immunohistochemistry results showed reduced mTOR expression in the pancreas during ZOTC treatment. Conclusively, the results obtained suggest that ZOTC treatment against fructose-STZ-induced type 2 diabetes rat models can help regulate blood glucose, insulin levels, and normalize pancreatic β cell damage. PRACTICAL APPLICATIONS: Type 2 diabetes is a chronic metabolic disorder that affects a large number of populations worldwide. Zingiber officinale (ZO) and Terminalia chebula (TC) has been used in traditional medicine since ancient times against various ailments, including diabetes. In this study, we reported the effect of the combined ZOTC that showed significant blood glucose reduction and increased insulin levels via mTOR when compared to individual treatments. This finding is valuable for food technologists and alternative medicine practitioners to know the antidiabetic effect of the ZOTC combination.

The effect of UV radiation in the presence of TiO2-NPs on Leishmania major promastigotes

Background: Cutaneous leishmaniasis is a parasitic disease, which is difficult to treat due to high drug resistance and adverse side effects. Photodynamic therapy by ultraviolet radiation using materials with high photocatalytic features like titanium dioxide nanoparticles (TiO2-NPs) is an emerging treatment for this disease. In this study, TiO2-NPs with ultraviolet (UV) radiation were administered as photodynamic therapy against Leishmania Major (LM) promastigotes.Methods: Two forms of TiO2 viz. including Anatase and Rutile were administered in two UV ranges< UVA and UVB for different time periods (30 and 60 min). Finally, 24 and 48 h after incubation, the MTS test was performed and cell survival percentage was calculated.Results: The mean size of Anatase and Rutile-NPs is approximately 32.5 and 50.9 nm respectively by DLS and FE-SEM, and crystal phase is emphasized by XRD. The combined treatment of LM with TiO2-NPs and UV has significant effects on LM promastigotes, which vary depending on NP and UV types. The synergistic effect was anticipated in the groups irradiated by UV-B in the presence of Rutile NPs.Conclusion: The combined treatment with UV- radiation and TiO2-NPs can be effective in killing the promastigotes of Leishmania major. The proper concentration of NPs and the type of UV-radiation must be taken into consideration. The results suggest improved treatment methods, after proper in vivo studies.

Necroptosis of osteoblasts was induced by breast cancer cells in vitro.

BackgroundBone metastasis of breast cancer could lead to serious osteolysis and severe pain. This study is aimed to investigate the existence of necroptosis, a new type of programmed cell necrosis pathway, in breast cancer-induced osteoblast cell death.MethodsIn this study, conditioned medium (CM) of breast cancer cells was prepared to simulate the micro-environment of bone metastasis in breast cancer in vitro and co-cultured with osteoblast. Then the percentage of cell survival and death was detected via cell viability and flow cytometry. Western blot and PCR were taken to measure protein and mRNA expression level of RIPK 3, MLKL and caspase 3 respectively.ResultsCM could induce osteoblasts death, including apoptosis and necroptosis and necrostatin-1 plus Z-IETD-FMK could decrease the percentage of death cells significantly in the flow cytometry detection. Moreover, CM could increase cleaved caspase 3, RIPK 3 and p-MLKL significantly, while RIPK 3 and p-MLKL was reduced statistically when osteoblasts were treated with Necrostatin-1 (Nec-1). In addition, the mRNA level of three proteins was not consistent with the change of their corresponding protein level.ConclusionsIn conclusion, the necroptosis pathway exists in osteoblast cell death pathway induced by breast cancer cells and could be inhibited by Necrostatin-1 (Nec-1).

Amylase concentration and activity in the amniotic fluid of fetal rats with retinoic acid induced myelomeningocele

Background In utero neurologic injury in myelomeningocele (MMC) occurs via a two-hit process: failed neural tube closure followed by neurodegeneration in utero. Meconium in the amniotic fluid contains pancreatic digestive enzymes and is neurotoxic in rat models of MMC. Objectives The objectives of this study were to demonstrate the neurotoxicity of α-amylase and to compare the enzyme concentration and activity in the amniotic fluid of rats with retinoic acid induced MMC to a healthy control population. Study design Timed pregnant Sprague Dawley rats were gavage fed all-trans retinoic acid (60 mg/kg) in olive oil on gestational day E10 to induce a MMC defect. Control rats received olive oil. Amniotic fluid was collected on embryonic days E15, E17, E19, and E21. The amniotic fluid amylase concentration and relative activity were measured at each gestational age, and levels were compared between the MMC and control groups using Wilcoxon Rank Sum and Kruskal–Wallis tests. In a subset of dams sacrificed on E10.5, neuroepithelial cells were isolated from control embryos and exposed to α-amylase in increasing concentrations. Percentage of cell survival was assessed with CellProfiler software. Results Amniotic fluid amylase activity for embryonic days E15, E17, E19, and E21 was determined for MMC and control pups. Amylase activity increased significantly from E15 to E21 in both control (p = 3.0 × 10−5) and MMC (p = 1.5 × 10−5) groups. Relative amylase activity was significantly increased in MMC pups compared to controls on E19 (247,792.8 versus 106,263.6; p = .0019) and E21 (772,645.8 versus 481,975.3; p = .021); no difference was detected on E15 (36,646.8 versus 40,179.3; p = .645) or E17 (121,617.5 versus 71,750; p = 1.000). In vitro, amylase demonstrated dose-dependent toxicity to fetal rat neuroepithelial cells. Conclusion Amylase concentration and activity level were higher in the amniotic fluid of rats with retinoic acid induced MMC compared to controls with advancing gestational age. As amylase is toxic to neural epithelial cells, the higher activity of this digestive enzyme in fetuses with MMC may be a contributor to neural tube damage in utero. Future research should focus on amylase and other digestive enzymes in human MMC, as they may serve as potential targets of in utero therapy.

Antioxidant, Antimicrobial and Cytotoxic Potential of &amp;lt;i&amp;gt;Abelmoschus esculentus&amp;lt;/i&amp;gt;

Abelmoschus esculentus is an important medicinal plant belongs to family Malvaceae. It originates from Ethiopia and is widely spread all over tropical, subtropical and warm temperate regions of the world. This research work has been designed to evaluate the antioxidant, antimicrobial and toxicological potential of A. esculentus leaves and seeds. The antifungal and antioxidant components of A. esculentus leaves and seeds were extracted by using four solvent systems (80% methanol, 80% ethanol, 100% methanol and 100% ethanol) and leaves presented maximum extract yield (38.1 g/100g DW)in 80% methanolic solvent system. Phytochemical analysis of A. esculentus leaves and seeds extracts performed in terms of total phenolic and total flavonoid contents, showed that 80% methanolic leaves extract offered highest total phenolic contents (31.2 mg GAE/g DW), whereas 80% ethanolic leaves gave maximum total flavonoid contents (41.8 mg CE/g DW). Antioxidant activity was determined by DPPH radical scavenging activity and measure of reducing power. Results revealed that 80% methanolic leaves extract showed highest radical scavenging activity and reducing potential. Antimicrobial activity of A. esculentus leaves and seeds was investigated by Disc Diffusion Method and Minimum Inhibitory Concentration (MIC). Results showed that 80% methanolic extract of leaves exhibited highest antibacterial and antifungal potential against P. multocida (30 mm DIZ) and A. paraciticus (29 mm DIZ), respectively. Cytotoxicity analysis was performed on BHK-21 cell by adopting the MTT assay. The cytotoxicity activity of the 80% methanolic extract of leaves was evaluated by noticing the cell survival percentage (52.5%). Overall results of the present study showed that 80% methanolic leaves extracts of A. esculentus possesses very good antioxidant, antimicrobial and cytotoxic properties.

Streptococcus mutans isolated from children with severe-early childhood caries form higher levels of persisters

ObjectivesDental caries is the most common chronic infectious disease in children. Streptococcus mutans, the main cariogenic bacterial species, produces persisters, nongrowing dormant variants of regular cells associated with chronicity of diseases. We hypothesized that the recurrent nature of caries, particularly within populations with high-caries risk, is due partly to specific phenotypic features of S. mutans such as its ability to form persisters. We aimed to investigate the genotypic and phenotypic differences between the S. mutans from children with severe early-childhood caries (S-ECC) and those without caries. MethodsS. mutans from plaque samples of caries-free (CF) and S-ECC children were tested for their ability to adapt to a lethal pH in an acid tolerance response assay. The persister levels of S. mutans isolates was quantified in both groups. ResultsS. mutanswas identified in all 23 S-ECC but only 6 of the 21 CF subjects. In most subjects, only one dominant S. mutans genotype was detected. No statistically significant differences in the mean survival percentage of S. mutans were observed between the two groups at a lethal pH of 3.5. However, the dominant genotype within a particular S-ECC subject exhibited a higher percentage of cell survival compared to those in the CF group. In S-ECC patients, S. mutans isolates displayed a ∼15-fold higher persistence phenotype than S. mutans isolates from CF patients. ConclusionsThe ability of S. mutans to produce high levels of persisters may contribute to part of an individual’s ability to control caries disease activity and recurrent lesions.

Protective effect of a protease inhibitor from Agaricus bisporus on Saccharomyces cerevisiae cells against oxidative stress

Protease inhibitors are known to resist damage to host organisms against external threats, hence form a part of their defense system. This property of protease inhibitors was studied on protecting oxidatively stressed Saccharomyces cerevisiae yeast cells. The protease inhibitor was extracted from Agaricus bisporus, an edible mushroom. The inhibitor showed the presence of antioxidant activity as the purified inhibitor fraction gave an IC50 value of 45.13 ± 0.88 µg/mL and 33.30 ± 1.5 µg/mL when checked, respectively, by 2, 2-diphenyl-1-picrylhydrazyl, DPPH and 2, 2′-azo-bis(3-ethylbenzthiazoline-6- sulfonic acid), ABTS•+ scavenging activity. The yeast cells’ survival rate (%), was determined through 3-(4, 5-dimethylthiazol-2-yl) - 2, 5-diphenyltetrazolium bromide, MTT assay, and it was found that in the presence of 2 mM H2O2 cell survival decreased to 26.33%, whereas when the experiment was conducted in the presence of protease inhibitor and 2 mM H2O2 cell survival percentage rose to 74%. The protease inhibitor’s effect on the oxidatively stressed yeast cells was further studied by using Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM) and Confocal Microscopy to understand the morphological changes. The viable and non-viable cell populations were quantified using Fluorescence Assorted Cell Sorting (FACS) using propidium iodide, PI, 4′, 6-diamidino-2-phenylindole, DAPI and 2′, 7′-dichlorofluorescein, DCF dyes.

Utilizing photosensitizing and radiosensitizing properties of TiO2-based mitoxantrone imprinted nanopolymer in fibrosarcoma and melanoma cells.

Some materials such as TiO2 display a luminescence property when exposed to X-ray radiation. Therefore, a proper photosensitizer can induce photodynamic effects by absorbing the emitted photons from these materials during radiotherapy. In this way, the problem of limited photo- penetration depth in photodynamic therapy is resolved. In this paper, following the production of a nanopolymer containing TiO2 cores and imprinted for mitoxantron (MIP), the possibility of utilizing its optical and radio properties on two lines of cancer cells were studied. Mitoxantron (MX) was selected as the photosensitizer. The emission spectrum of the nanopolymers synthesized with/without MX was recorded during excitation by 6 MV X-rays. Also, the fluorescence signal of hydroxyl radicals produced into terephthalic acid medium by the nanopolymers were recorded during X irradiation. The percentage of cell survival following irradiation by X-rays was determined for various concentrations of drug agents by MTT assay. The synergistic index and IC50 were calculated to compare the findings. The emission spectrum of the nanopolymer reloaded with MX during X-ray irradiation indicated a considerable decline in comparison with the nanopolymer without MX. The level of free radicals produced by nanopolymer was significantly increased during irradiation with X-rays. The highest mean of synergistic indexes was observed in MIP. The higher level of hydroxyl free radicals in MIP and lower cell viability in the DFW cell line as well as enhanced treatment efficiency confirm the hypothesis regarding the production of photodynamic effects by synthesized nanopolymer during radiotherapy.

The Evaluation of Optimum Radiation Dose to Prevent Transfusion-Associated Graft-Versus-Host Disease Among the Iranian Population

Background: Transfusion-associated graft-versus-host disease is a complication being mortal in 90% of cases. Today, Co-60 devices are utilized for the suppression of the lymphocytes to prevent this complication. Due to problems such as radioactive source age, problems in source replacement, and protection risks, we sought to replace a linear accelerator for this purpose and carefully determinate the X-ray dose and other settings. Methods: First, the venous blood of the right-handed people with blood group O+ was exposed to different radiation doses of Linac and diluted. The proliferative responses of the exposed cells were examined by MTT assay and TB in comparison with controls after 48h. The average percentage of cell survival at each delivered dose and the required dose of radiation for the inhibition of lymphocyte proliferation were checked and the optimal dose of radiation was obtained as 25 Gy in a special radiation condition. Results: The results indicated that the irradiation with 25 Gy at 3.5 Gy/min in the internal mid-plan by compact 6MV for 30*30-field size at an 8 cm-distance of the samples to the source and 1370-monitor unit could completely suppress the lymphocytes. Under the same condition and 1096-monitor unit for total 20 Gy, we did not find optimal results. Conclusions: In this research, we found that a linear accelerator in a specific exposure condition is a suitable alternative for Cobalt-60 sources and the above-mentioned condition of irradiation can be used to overcome the GVHD concern.

Establishment of incubation conditions to optimize the in vitro formation of mature Listeria monocytogenes biofilms on food-contact surfaces

Most microbial populations in food industry environments grow as adhered communities with subsequent biofilm formation on different food-contact surfaces, causing possible cross-contaminations to food products and posing a relevant risk to public health. The aim of this study was to develop a laboratory scale model to optimally form Listeria monocytogenes biofilms in their mature stage on stainless steel surfaces. The results showed that the best initial incubation period is 48 h when compared with 24 h and 72 h. The maximum growth for biofilm formation in the in vitro conditions tested was obtained after a week of incubation with different series of washing and nutrient renewal, with a total cell amount of 7.25 log CFU cm−2 and a cell survival rate of 94.47%. Significant differences were obtained (P < 0.05) when 48 h as the total incubation period was compared with 48 h + 24 h, 48 h + 48 h and 48 h + 24 h + 24 h. These conditions were also compared with each other and no statistical differences were found (P = 0.5392, P = 0.5542, P = 0.9965, respectively). When the same conditions were compared with the one-week incubation, significant differences were obtained (P < 0.001) in all the cases except for the condition of 48 h + 24 h + 24 h + 72 h + 24 h (P = 0.1654), but they presented a lower count and cell survival percentage. The incubation period, the series of washes and the renewal of nutrients directly influences the extracellular matrix production, demonstrating that a one-week incubation period is the most suitable condition for producing the proteins and carbohydrates that give the biofilm system consistency and robustness. The effect of the conditioned protein layer demonstrated an 8–13% greater biofilm formation of the strains CECT 5366, 5672 and 5873 on the polystyrene surfaces, but no significant differences were found in microbial counts (P > 0.05). The proposed in vitro model to form biofilms in their mature stage is completely crucial to understanding how to reproduce and eliminate them not only in vitro, but also in real conditions.