MTT Technique Research Articles

Surface-modified dendrimer nanoconjugate for targeting delivery of rifampicin

ObjectiveThe study aimed to formulate and evaluate the efficiency of surface-modified Polyamidoamine dendrimer (PAMAM) as a targeted nanocarrier for the antimycobacterial agent rifampicin. MethodsThe periphery of dendrimer was modified with mannose using α-D-mannopyranosylphenyl isothiocyanate and characterized using analytical techniques i.e., infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR). Then, rifampicin was encapsulated into mannosylated dendrimers and evaluated for size, shape, encapsulation efficiency (EE%), drug-dendrimer interaction, and drug release. The toxicity of the nanoconjugates were assessed towards macrophages using the MTT technique. The study investigated the internalization of dendrimer nanoparticles into macrophages to assess the impact of mannose conjugation. Dendrimers were fluorescently labelled and the internalization was quantified using a fluorescence spectroscopy and flow cytometer methods. ResultsMannosylated dendrimers with different mannose densities were successfully developed. The nanoparticles were within the nanoscopic scale. The dendrimers appeared spheroidal under scanning electron microscopy (SEM), and thermal analysis confirmed the conjugation of rifampicin into the dendrimers. The nanoparticles exhibited a good EE% for rifampicin at 10.34 % w/w. After surface mannosylation, the EE% further increased, ranging from 43.43 % to 57.91 % w/w. Mannose-functionalized dendrimers prolonged rifampicin release in physiological pH, while a rapid release in pH 4.5 medium was noticed. The cytotoxicity of the dendrimers in RAW macrophages was considerably reduced after mannose functionalization. In vitro studies indicated that mannose significantly increased the macrophage uptake of nanoconjugates, likely via lectin receptor-mediated endocytosis. ConclusionOverall results suggested mannosylated dendrimers as an effective targeted pulmonary delivery system for rifampicin with negligible cytotoxicity.

New Chalcone Incorporated Structurally Modified Pyridine-Pyrimidine Derivatives as Anticancer Agents: Their Design, Synthesis, and in-vitro Evaluation.

Chalcone-incorporated pyridine-pyrimidines i.e. derivatives of (5-(6-(pyrimidin-5-yl)pyridin-3-yl)thiophen-2-yl)prop-2-en-1-one were synthesized and their structures were confirmed by analytical techniques. In addition, all the derivatives were examined for their capacity to fight against cancer towards four cell lines, including breast (MCF-7), prostate (DU-145 and PC3), and lung (A549) by utilizing the MTT technique and the clinically used chemotherapy medication, etoposide serving as a positive reference. All these results were expressed in IC50 μM, and values of synthesized compounds are compared with a reference drug, showing values ranging from 1.97±0.45 μM to 3.08±0.135 μM. Among those, a few compounds 10(a-e) demonstrated strong activities with corresponding cell lines.

Hydroalcoholic Extract of Fumaria Parviflora L. Can Potentially Detoxificate HepG2 Cell Line Following Carbon Tetrachloride Exposure

Background: Fumaria Parviflora L. (FP) is a well-known herb in Iranian traditional medicine, widely used for gastrointestinal diseases. Additionally, the detrimental effects of carbon tetrachloride (CT) on the liver and kidneys have been scientifically established. Objectives: This study was designed to assess the therapeutic effects of FP on HepG2 cells exposed to CT toxicity. Methods: After treatment with hydroalcoholic extract of the FP plant and/or CT, cell viability (using the MTT technique and lactate dehydrogenase assay) and apoptosis (using the diphenylamine assay and Annexin V/PI staining) were investigated. The ratio of BCL2/BAX mRNA expression levels was measured using real-time PCR. Data were analyzed using SPSS software. Results: A significant (P < 0.05) reduction in cell viability was observed at doses of 100 and 200 µg/mL of FP in HepG2 cells. The decreased survival rate following CT exposure was concentration-dependent and significant (P < 0.05). The IC50 of CT was determined to be 1.05 μg/mL for 24 hours. Effective detoxification associated with CT was found at 6.25 µg/mL in both the pre-treatment and simultaneous treatment groups. Cell apoptosis was significantly (P < 0.05) decreased following FP administration in CT-treated cells, with an increase in the ratio of BCL2/BAX mRNA expression levels. Conclusions: The hydroalcoholic extract of FP demonstrated hepatoprotective effects on CT-treated HepG2 cells.

Synthesis, crystal structure, Hirshfeld surface analysis, DFT studies and biological evaluation of bis(acetonitrile)bromido(η6-1-isopropyl-4-methylbenzene)osmium(II) hexafluoroantimonate

The complex bis(acetonitrile)bromido(η6-1-isopropyl-4-methylbenzene)osmium(II) hexafluoroantimonate, [OsBr(C10H14)(CH3CN)2]SbF6 (1) crystallizes as a triclinic system in space group P1 (no. 2), consisting of two cationic osmium(II) complex ions and two anionic hexafluoroantimonate counterions in each cell. Density Functional Theory (DFT) calculations were performed at the B3LYP/LanL2DZ and B3LYP/Def2 − SVP/Def2 − TZVP levels of theory. Reactivity descriptors and charge transfer properties were computed using frontier molecular orbital analysis. The DFT-optimized model systems from both basis sets were comparable to the experimentally determined X-ray diffraction results. Hirshfeld surface analysis shows significant non-covalent interactions including hydrogen bonds and halogen interactions. The MTT technique was utilized to conduct an in vitro cytotoxicity assay of 1 against cancerous cell lines (MCF-7 and A549) and normal cells (HEK293). Complex 1 is potent towards cancer cells as evidenced by the IC50 values of 5.02 ± 0.05 µM for MCF-7 cells and 13.36 ± 0.04 µM for A549 cells. Complex 1 demonstrated lower toxicity to normal cells with an IC50 value of 26.76 ± 0.08 µM for the HEK293 cell line compared to the doxorubicin standard with an IC50 value of 1.2 ± 0.1 µM.

Development of carboxymethyl cellulose/gelatin hydrogel loaded with Omega-3 for skin regeneration.

Hydrogels have several characteristics, including biocompatibility, physical similarity with the skin's extracellular matrix, and regeneration capacity. Cell migration and proliferation are facilitated by natural polymers such as gelatin (Gel) and carboxymethyl cellulose (CMC). Gelatin dressing acts as a structural framework for cell migration into the wound area, stimulating cell division and promoting granulation tissue formation. Omega-3 fatty acids from fish oil may prevent wound infection and improve the healing of wounds in the early stages. We studied the preparation of wound dressing containing Omega-3 and its ability to heal wounds. In this study, CMC-Gel hydrogels containing different concentrations of Omega-3 were investigated in full-thickness wounds. After the fabrication of the hydrogels by using surfactant (tween 20) and microemulsion method (oil in water), various tests such as SEM, Water uptake evaluation, weight loss, cell viability, blood compatibility, and in vivo study in rat cutaneous modeling during 14 days were performed to evaluate the properties of the fabricated hydrogels. The analysis of the hydrogels revealed that they possess porous structures with interconnected pores, with an average size of 83.23 ± 6.43μm. The hydrogels exhibited a swelling capacity of up to 60% of their initial weight within 24h, as indicated by the weight loss and swelling measurements. Cell viability study with the MTT technique showed that no cytotoxicity was observed at the recommended dosage, however, increasing the amount of omega-3 caused hemolysis, cell death, and inhibition of coagulation activity. An in vivo study in adult male rats with a full-thickness model showed greater than 91% improvement of the primary wound region after 2 weeks of treatment. Histological analysis demonstrated Omega-3 in hydrogels, which is a promising approach for topical skin treatment to prevent scar, and has shown efficacy as wound dressing by improving the repair process at the defect site.

Integrating Natural Deep Eutectic Solvents into Nanostructured Lipid Carriers: An Industrial Look.

The industries are searching for greener alternatives for their productions due to the rising concern about the environment and creation of waste and by-products without industrial utility for that specific line of products. This investigation describes the development of two stable nanostructured lipid carriers (NLCs): one is the formulation of a standard NLC, and the other one is the same NLC formulation associated with a natural deep eutectic solvent (NaDES). The research presents the formulation paths of the NLCs through completeness, which encompass dynamic light scattering (DLS), zeta potential tests, and pH. Transmission electron microscopy (TEM) and confocal microscopy were performed to clarify the morphology. Cytotoxicity tests with zebrafish were realized, and the results are complementary to the in vitro outcomes reached with fibroblast L132 tests by the MTT technique and the zymography test. Infrared spectroscopy and X-ray diffractometry tests elucidated the link between the physicochemical characteristics of the formulation and its behavior and properties. Different cooling techniques were explored to prove the tailorable properties of the NLCs for any industrial applications. In conclusion, the compiled results show the successful formulation of new nanocarriers based on a sustainable, eco-friendly, and highly tailorable technology, which presents low cytotoxic potential.

Investigating the effect of LncRNA DLGAP1-AS2 suppression on chemosensitivity of gastric cancer to chemotherapy.

Gastric cancer, as the fifth most frequent disease and the fourth foremost cause of cancer-related death worldwide, remains a main clinical challenge due to its poor prognosis, limited treatment choices, and ability to metastasize. Combining siRNAs to suppress lncRNA with chemotherapeutic medications is a novel treatment approach that eventually increases the therapeutic efficacy of the drug while lessening its adverse effects. This study was performed with the purpose of examining the impact of inhibiting DLGAP1-AS2 expression on gastric cancer cells' drug chemosensitivity. AGS cells were cultured as the study cell line and were transfected with an optimum dose of DLGAP1-AS2 siRNA and then treated with oxaliplatin. Cell viability was examined using the MTT technique. Apoptosis and cell cycle were evaluated using Annexin V/PI staining and flow cytometry. Later, the scratch test was conducted to investigate the ability of cells to migrate, and the inhibition of the stemness of AGS cells was further investigated through the colony formation method. Finally, the qRT-PCR technique was used to assess the expression of Bax, Bcl-2, Caspase-3, p53, MMP-2, and CD44 genes. The MTT test indicated the effect of gene therapy with siRNA and oxaliplatin in combination reduced the chemotherapy drug dose to 29.92µM and increased AGS cells' sensitivity to oxaliplatin. Also, the combination therapy caused a significant increase in apoptosis. However, it reduced the stemness feature, the rate of cell viability, proliferation, and metastasis compared to the effect of each treatment alone; the results also showed the arrest of the cell cycle in the Sub G1 phase after the combined treatment and a further reduction in the number and size of the formed colonies. Suppressing the expression of lncRNA DLGAP1-AS2 by siRNA followed by treatment with oxaliplatin can be utilized as an effective and new therapeutic technique for gastric cancer therapy.

In vitro cytotoxicity and apoptosis inducing effect of Vallarai Kirutham with Rasa Parpam in HeLa Cell Lines

Cancer is an uncontrolled proliferation of cells that can affect nearby tissues as well as distant organs. Cervical cancer is predicted to be among the four most prevalent cancers in women overall in 2020, with 342,000 mortality and 604,000 new cases. Around 6-29% of all cancers in women in India are caused by cervical cancer. Women who have had several sexual partners, recurrent abortions, or vaginal deliveries—all of which cause repetitive stress to the cervix are more likely to develop cervical cancer. Radiation, chemotherapy, and surgery are being used to treat cancer. All of these treatments come with side effects, which can range in severity and have a serious psychological impact on patients. The Siddha system has a number of accessible herbo mineral formulations for treating cervical cancer. According to Siddha literature (Agasthiyar Vaidhya Rathina Churukkam, 360), the treatment of cervical cancer (yoni puttru) is advised for the Siddha medicine Vallarai Kirutham with Rasa parpam. The goal of this research was to find out this formulation's potential anti-cancer properties using MTT assays on HeLa cell lines. These findings of the current study suggest that the MTT technique's lowest reading for cell viability was 1.26±0.009% at a concentration of 100 µl/ml. This was followed by concentrations of 75 µl/ml, 50µl/ml and 25µl/ml which showed 7.91±0.004%, 23.91 ±0.01% and 37.17± 0.01% similarly, 10 µl/ml shows 52.09±0.01%.The matching IC50 value was discovered to be 12.44% and AO/EB dual staining was used to examine apoptotic activity. The outcomes of in-vitro experiments using the HeLa cell line. According to the outcome of in vitro experiments done on the HeLa cell line, the drug had substantial anti-cancer and therapeutic value in the management of cervical tumours .With a variety of time tested medications the Siddha system has shown outcomes that were clinically important for the management of cervical carcinoma. One of those medications has to undergo additional testing in patients with cervical cancer through clinical research.

Activity of zinc oxide and zinc borate nanoparticles against resistant bacteria in an experimental lung cancer model.

Recent research indicates a prevalence of typical lung infections, such as pneumonia, in lung cancer patients. Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii stand out as antibiotic-resistant pathogens. Given this, there is a growing interest in alternative therapeutic avenues. Boron and zinc derivatives exhibit antimicrobial, antiviral, and antifungal properties. This research aimed to establish the effectiveness of ZnO and ZB NPs in combating bacterial infections in lung cancer cell lines. Initially, this study determined the minimal inhibitory concentration (MIC) and fractional inhibitory concentration (FIC) of zinc oxide nanoparticles (ZnO NPs) and zinc borate (ZB) on chosen benchmark strains. Subsequent steps involved gauging treatment success through a lung cancer-bacteria combined culture and immunohistochemical analysis. The inhibitory impact of ZnO NPs on bacteria was charted as follows: 0.97µg/mL for K. pneumoniae 700603, 1.95µg/mL for P. aeruginosa 27853, and 7.81µg/mL for Acinetobacter baumannii 19,606. In comparison, the antibacterial influence of zinc borate was measured as 7.81µg/mL for Klebsiella pneumoniae 700603 and 500µg/mL for both P. aeruginosa 27853 and A.baumannii 19606. After 24h, the cytotoxicity of ZnO NPs and ZB was analyzed using the MTT technique. The lowest cell viability was marked in the 500µg/mL ZB NPs group, with a viability rate of 48.83% (P < 0.001). However, marked deviations appeared at ZB concentrations of 61.5µg/mL (P < 0.05) and ZnO NPs at 125µg/mL. A synergistic microbial inhibitory effect was observed when ZnO NP and ZB were combined against the bacteria under investigation.

Determination of the effects of irisin hormone in SKMEL-30 cells

Melanocytes, the skin's pigment-producing cells, are the source of the skin cancer known as melanoma. Numerous variables, including as immune system interactions, tumor microenvironment, and genetic alterations, have an impact on the development and behavior of melanoma. The purpose of this study was to ascertain the impact of irisin on melanoma cells. The molecular effects of irisin SKMEL-30 on human melanoma cancer cells were examined for this aim. By using MTT technique, the effects of irisin on cell growth were examined. Real-time polymerase chain reaction was used to examine changes in gene expression level. The concentrations of sialic acid were measured using spectrophotometry. In the investigation, the irisin IC50 value for a 24-hour application was determined to be 30 nM. In comparison to the control group, sialic acid levels in the irisin-treated group of SKMEL-30 cells were significantly lower. In the qRT-PCR investigation, ST8SIA-2, one of the glycosyltransferase genes, increased 12.591-fold in the application group whereas cas8, one of the apoptotic genes, increased 82.481-fold. In conclusion, flow cytometry analyses proved that administration of 30 nM irisin to SKMEL-30 cells influences cell proliferation but does not cause apoptosis. It was shown that sialic acid substitution reduced the proliferative and metastatic potential of SKMEL- 30 cells.

Study the Effect of Calendula officinalis Extract Loaded on Zinc Oxide Nanoparticle Cream in Burn Wound Healing.

The skin, the body's largest organ, acts as a protective barrier against pathogens and environmental damage. Skin burns can result from heat, chemicals, friction, or electricity. Nanoscience has recently been utilized to create ointments and creams for burns. Zinc oxide nanoparticles are crucial due to their antimicrobial and antioxidant properties. In this study, a cream containing nanoparticles was loaded with calendula extract, and its ability to promote tissue healing was investigated in Wistar rats with skin burns. The zinc oxide nanoparticles were chemically synthesized and loaded with calendula extract. The morphology and physicochemical properties of the nanoparticles were confirmed by SEM, ZETA size, XRD, and FTIR assays. The MTT technique was employed to assess the cream's impact on fibroblast growth. The antimicrobial activity of the nanoparticles was investigated against Pseudomonas using the MIC method. Real-time PCR was used to determine the expression of the Bax and Bcl-2 genes in aeruginosa. The results showed that zinc oxide nanoparticles at high concentrations increased the proliferation of the fibroblast cells. Histopathological studies showed granulation and epithelialization of the tissue without any hemorrhage or tissue infection during the first days of treatment with this cream. The animal models treated with the cream showed an increase in Bcl-2 gene expression and a decrease in Bax expression. We concluded that zinc oxide nanoparticles loaded with calendula extract have a practical effect in healing burn wounds due to their unique antibacterial properties of zinc oxide nanoparticles and their anti-inflammatory and wound-healing effects. The synergistic effect of these two substances significantly improved the healing process. This newly developed cream can be introduced as a successful and viable treatment option in burn wounds.

Rapid synthesis of Silver Nanoparticles by Rheum ribes L Fruit Peels Anticancer and Antimicrobial Effects with Biocompatible Structures

Silver nanoparticles (AgNPs) are useful substances with a wide range of uses. utilizing the extract taken from the peels of the Rheum ribes L.(Rr) fruit growing in the Erzurum region, silver nanoparticles were speedily created in this study utilizing a quick, easy, and environmentally friendly technique without harmful stages. In order to evaluate the attributes of the synthesized Rr-AgNPs, FE-SEM or TEM micrographs were utilized to characterize the morphology of the AgNPs. A UV-visible spectrophotometer was used to assess the highest absorbance bands of RR-AgNPs. These data were used to produce RR-AgNPs, which were characterized as having exclusively negative surface charges of -25 mV, a spherical shape, maximum absorbance at 428 nm wavelength, and 96 nm size distribution. The effectiveness of the produced AgNPs for usage in medical applications was assessed using the MTT technique with microdilution. Minimum Inhibition Concentrations of Rr-AgNPs on pathogen strains were found to range from 0.03-0.50 mg/L. Additionally, it was discovered that AgNPs effectively suppressed malignant cells in the investigation for the anticancer effects of AgNPs, with rates of 86.27%, 74.67%, and 73.49%. Healthy cells were not subjected to any inhibitory effects at the same concentrations that caused this result.

Nutritional Values and Their Potential Applications in Food Products of Krabok Seed (Irvingia malayana)

This investigation examined the nutritional value, mineral content, total phenolic content, antioxidant activity and cytotoxicity of uncooked and cooked Krabok seeds. Proximate analysis was carried out to determine the nutrient composition of uncooked and cooked Krabok seeds. The average fat, protein, carbohydrate, moisture, ash, and fiber contents were found to be 75.42, 11.78, 6.72, 2.68, 1.85 and 1.54 %, respectively. The Folin-Ciocalteu method was employed for the quantitative analysis of total phenolic compounds. All 6 samples of Krabok seeds extracted, both cooked and uncooked, were extracted using hexane, ethyl acetate, and ethanol, and subjected to the test. All the extracted samples contained phenolic compounds, with amounts ranging from 29.10 to 130.26 µg GAE/mL. The highest levels of phenolic compounds were observed in the ethanolic extract of cooked Krabok seeds. The antioxidant properties were evaluated using the DPPH assay. The scavenging activity of the extracted Krabok seed samples ranged from 9.84 to 62.31 %. The ethanolic extract of uncooked Krabok seeds exhibited the highest antioxidant activity. To assess the toxicity of the Krabok seed extract, HaCaT, Vero and RAW 264.7 cell liness were employed using the MTT technique. The results revealed that cell survival remained above 80 % for all doses of both uncooked and cooked Krabok seed extracts. These findings suggest that Krabok seeds are a valuable source of fat and other essential nutrients. The crude ethanolic extract exhibited a significant amount of phenolic content and antioxidant activity. Therefore, it could be utilized in the production of snacks or cosmetics. HIGHLIGHTS Krabok seeds has high nutrients and energy value. Krabok seeds has high phenolic compounds and non-toxic to cells. Krabok seeds were thought to be an inexpensive source of nutrients and a reliable supply of prospective culinary ingredients. GRAPHICAL ABSTRACT

Synthesis of New Bi-Triazoles with Plasmocide Action Against Plasmodium falciparum

Background: A series of bi-triazoles conjugates 1,2,3 and 1,2,4 was synthesized with an aim to study the evaluation of the antimalarial profile of families of triazole derivatives. The study used the W2 strain of Plasmodium falciparum (Chloroquine-Resistant), to determine the inhibitory concentration of 50% of the parasites (IC50) and HepG2 cells to describe the cytotoxic concentration for 50% of the cells (CC50). Among the study classes, bi-triazoles stood out with IC50 values between 8.9 to 0.45 μM; highlighted the compound 14d (IC50 of 0.45 ± 0.02 μM) with the most promising result. Regarding the cytotoxic concentration, all compounds that presented IC50 values ≤ 100 μM were evaluated. Three compounds stood out as the highest selectivity index (SI) values, 14b (SI ˃111.1), 13d (SI ˃111.1) and 14d (SI ˃1.111). Such results expose the importance of working with classes of molecules that allow rapid synthesis and dispositions for structural changes. Highlighting the evolution of the IC50 values of the compounds, when adding the second triazole block. Thus, the results found in this study, have the possibility of choosing new molecules for the treatment of malaria. Objective: This work was to synthesize a series of bi-triazole conjugates 1,2,3 and 1,2,4-triazole moiety and evaluate their activities against Plasmodium falciparum. Methods: The bi-triazole was synthesized in a 3-step route in moderated yields, and their structures were confirmed by NMR spectral data analyses. For the in vitro antiplasmodial assays, the SYBR Green fluorimetric technique and the W2 strain were used, where an IC50 (Inhibitory Concentration) value was obtained for each compound. The compounds were also evaluated for their stagespecificity and speed of action (W2 strain). Safety tests were performed to determine the hemolytic and cytotoxic action of the evaluated compounds. In these tests, the cell lines HepG2 and VERO were used, and the cytotoxicity was evaluated by the MTT technique. This allowed the CC50 values to be obtained (Cytotoxic Concentration). Subsequently, the Selectivity Index (SI) was calculated for each compound. Results: The newly synthesized bi-triazole compounds could serve as potent leads for the development of novel antimalarial compounds. In general, the bi-triazoles with trifluoromethyl group present at 1,2,4-triazole moiety proved to be more potent regarding antiplasmodial activity. Conclusion: The synthesized bi-triazole compounds could serve as potent leads for the development of novel antimalarial agents.

Halogenated class of oximes as a new class of monoamine oxidase-B inhibitors for the treatment of Parkinson’s disease: Synthesis, biochemistry, and molecular dynamics study

Oximes are the promising structural scaffold for inhibiting monoamine oxidase (MAO)-B. Eight chalcone-based oxime derivatives were synthesized by microwave-assisted technique, and their ability to inhibit human MAO (hMAO) enzymes were tested. All compounds showed higher inhibitory activity of hMAO-B than hMAO-A. In the CHBO subseries, CHBO4 most potently inhibited hMAO-B with an IC50 value of 0.031 μM, followed by CHBO3 (IC50 = 0.075 μM). In the CHFO subseries, CHFO4 showed the highest inhibition of hMAO-B with an IC50 value of 0.147 μM. Compound CHBO4 had the highest selectivity index (SI) value of 1290.3. However, CHBO3 and CHFO4 showed relatively low SI values of 27.7 and 19.2, respectively. The -Br substituent in the CHBO subseries at the para-position in the B-ring showed higher hMAO-B inhibition than the -F substituent in the CHFO subseries. In both series, hMAO-B inhibition increased with the substituents at para-position in A-ring (-F > -Br > -Cl > -H in order). Compound CHBO4 (-F in A-ring and -Br in B-ring) was 12.6-times potent than the substituents-reversed compound CHFO3 (-Br in A-ring and -F in B-ring; IC50 = 0.391 μM). In the kinetic study, Ki values of CHBO4 and CHFO4 for hMAO-B were 0.010 ± 0.005 and 0.040 ± 0.007 μM, respectively, with competitive inhibitions. Reversibility experiments showed that CHBO4 and CHFO4 were reversible hMAO-B inhibitors. In the cytotoxicity test using the Vero cells by the MTT technique, CHBO4 had low toxicity with an IC50 value of 128.8 µg/mL. In H2O2-induced cells, CHBO4 significantly reduced cell damage by scavenging reactive oxygen species (ROS). Molecular docking and dynamics showed the stable binding mode of the lead molecule CHBO4 on the active site of hMAO-B. These results suggest that CHBO4 is a potent reversible, competitive, and selective hMAO-B inhibitor and can be used as a treatment agent for neurological disorders.

Green synthesis of silver nanoparticles mediated Diospyros kaki L. (Persimmon): determination of chemical composition and evaluation of their antimicrobials and anticancer activities.

The eco-friendly synthesis of metallic nanoparticles (MNPs) using biological materials is an encouraging and innovativeness approach to nanotechnology. Among other synthesizing methods, biological methods are chosen because of their high efficiency and purity in many aspects. In this work, using the aqueous extract obtained from the green leaves of the D. kaki L. (DK); silver nanoparticles were synthesized in a short time and simply with an eco-friendly approach. The properties of the synthesized silver nanoparticles (AgNPs) were characterized using various techniques and measurements. In the characterization data of AgNPs, Maximum absorbance at 453.34nm wavelengths, the average size distribution of 27.12nm, the surface charge of -22.4mV, and spherical appearance were observed. LC-ESI-MS/MS analysis was used to assess the compound composition of D. kaki leaf extract. The chemical profiling of the crude extract of D. kaki leaves revealed the presence of a variety of phytochemicals, predominantly phenolics, resulting in the identification of five major high-feature compounds: two major phenolic acids (Chlorogenic acid and Cynarin), and tree flavonol glucosides (hyperoside, quercetin-3-glucoside, and quercetin-3- D-xyloside). The components with the highest concentrations were cynarin, chlorogenic acid, quercetin-3- D-xyloside, hyperoside, and quercetin-3-glucoside, respectively. Antimicrobial results were determined by a MIC assay. The biosynthesized AgNPs exhibited strong antibacterial activity against the human and food pathogen Gram (+ and -) bacteria and good antifungal activity against pathogenic yeast. It was determined that 0.03-0.050μg/mL concentrations ranges of DK-AgNPs were growth suppressive concentrations on all pathogen microorganisms. The MTT technique was used to study the cytotoxic effects of produced AgNPs on cancer cell lines (Glioblastoma (U118), Human Colorectal Adenocarcinoma (Caco-2), Human Ovarian Sarcoma (Skov-3) cancer cell lines, and Human Dermal Fibroblast (HDF) healthy cell line). It has been observed that they have a suppressive effect on the proliferation of cancerous cell lines. After 48h of treatment with Ag-NPs, the DK-AgNPs were found to be extremely cytotoxic to the CaCo-2 cell line, inhibiting cell viability by up to 59.49% at a concentration of 50gmL-1. It was found that the viability was inversely related to the DK-AgNP concentration. The biosynthesized AgNPs had dose-dependent anticancer efficacy. Because of the high concentration of bioactive chemicals in Diospyros kaki, it may be employed as a biological resource in medicinal applications. DK-AgNPs were shown to be an effective antibacterial agent as well as a prospective anticancer agent. The results provide a potential approach for the biogenic production of DK-AgNPs utilizing D. kaki aqueous leaf extract.

The First Report of In Vitro Antifungal and Antibiofilm Photodynamic Activity of Tetra-Cationic Porphyrins Containing Pt(II) Complexes against Candida albicans for Onychomycosis Treatment.

Onychomycosis is a prevalent nail fungal infection, and Candida albicans is one of the most common microorganisms associated with it. One alternative therapy to the conventional treatment of onychomycosis is antimicrobial photoinactivation. This study aimed to evaluate for the first time the in vitro activity of cationic porphyrins with platinum(II) complexes 4PtTPyP and 3PtTPyP against C. albicans. The minimum inhibitory concentration of porphyrins and reactive oxygen species was evaluated by broth microdilution. The yeast eradication time was evaluated using a time-kill assay, and a checkerboard assay assessed the synergism in combination with commercial treatments. In vitro biofilm formation and destruction were observed using the crystal violet technique. The morphology of the samples was evaluated by atomic force microscopy, and the MTT technique was used to evaluate the cytotoxicity of the studied porphyrins in keratinocyte and fibroblast cell lines. The porphyrin 3PtTPyP showed excellent in vitro antifungal activity against the tested C. albicans strains. After white-light irradiation, 3PtTPyP eradicated fungal growth in 30 and 60 min. The possible mechanism of action was mixed by ROS generation, and the combined treatment with commercial drugs was indifferent. The 3PtTPyP significantly reduced the preformed biofilm in vitro. Lastly, the atomic force microscopy showed cellular damage in the tested samples, and 3PtTPyP did not show cytotoxicity against the tested cell lines. We conclude that 3PtTPyP is an excellent photosensitizer with promising in vitro results against C. albicans strains.

Studying the effect of gene fusion of A and C types capsular synthesizing enzymes and anticancer sequence on inducing the expression of apoptotic BCL-2, BAX, and Caspase-3 genes by Real-time RT-PCR method

BackgroundToday, uterine cancer is one of the most important causes of death in the world and is one of the major problems in human health. There have been numerous reports of the effect of Streptococcus agalactiae peptide and capsular products against cancer cell lines. Objective: This study aimed to research recombinant peptide CPSA-CPSC-L-ACAN and investigate its apoptotic effect against the HeLa cell line by Real-Time-RT PCR. DesignIn this study confirmation of the recombinant fusion peptide was performed by Western blotting. The effect of cytotoxicity of different concentrations of recombinant fusion peptide against the HeLa cell line was investigated by the MTT technique. The expression of apoptotic genes including BAX, BCL-2, and Caspase-3 in comparison with the GAPDH reference gene before and after exposure to recombinant fusion peptide was measured by Real-Time RT-PCR. ResultsRecombinant fusion peptide at a concentration of 63 μg/ml destroyed 50% of the HeLa cell line in 24 h and cell treatment with this concentration increased gene expression of Caspase-3 genes by 16 times, bax by 6 times and decreased the expression of bcl-2 by 0.176 times. ConclusionsThe results showed that treatment of the HeLa cell line with recombinant fusion peptide induced an apoptotic effect. The recombinant fusion peptide could probably help the medical community as a prophylactic or therapeutic treatment for cervical cancer.

Novel Bromo and methoxy substituted Schiff base complexes of Mn(II), Fe(III), and Cr(III) for anticancer, antimicrobial, docking, and ADMET studies

In this study, four new Mn(II), Fe(III), and Cr(III) complexes with two Schiff base ligands namely, 4-bromo-2-[(E)-{[4-(2-hydroxyethyl)phenyl]imino}methyl]phenol (HL1) and 2-[(E)-{[4-(2-hydroxyethyl)phenyl]imino}methyl]-4-methoxy phenol (HL2) have been synthesized and characterized. Different analytical and spectral methods have been used to characterize the ligands and their complexes. General formulas of [M(L)Cl2(H2O)2] for FeL1, CrL1 and CrL2, and [M(L)Cl(H2O)3] for MnL2 were proposed. HOMO and LUMO energies, as well as the electrical characteristics, have been calculated using DFT/B3LYP calculations with Gaussian 09 program. The optimized lowest energy configurations of the complexes are proven. The disc diffusion technique was used to test the pharmacological activities' antibacterial efficacy against diverse bacterial and fungus species. The MTT technique was used to assess the in vitro cytotoxicity of the ligands and their metal complexes on the Hep-G2 human liver carcinoma cell line and the MCF-7 human breast cancer cell line. All compounds displayed better activity compared to the free ligands. MnL2 complex showed predominant activity when compared to the other complexes with an IC50 value of 2.6 ± 0.11 μg/ml against Hep-G2, and against MCF-7 the IC50 value was 3.0 ± 0.2 μg/ml which is less than the standard drug cisplatin (4.0 μg/ml). UV–vis electronic spectrum and gel electrophoresis techniques have been used to investigate the compounds’ affinity to bind and cleavage CT-DNA. The interaction’s binding constants, or Kb, have been identified, and it was discovered that the new complexes' binding affinities are in the order of FeL1 > MnL2 > CrL2 > CrL1, and the binding mechanism has been suggested. To assess the kind of binding and binding affinity of the investigated drugs with human DNA, a molecular docking study was carried out (PDB:1bna). The acquired results supported the intercalation binding mechanism proposed in the experimental part and revealed that complexes may be inserted into the DNA molecule to stop DNA replication. According to ADMET data, the synthesized compounds have a high bioavailability profile and their physicochemical and pharmacological features remained within Lipinski's RO5 predicted limitations.

The Therapeutic Effects of Curcumin-coated Gold Nanoparticle Against Leishmania Major Causative Agent of Zoonotic Cutaneous Leishmaniasis (ZCL): An In Vitro and In Vivo Study

We synthesized and characterized curcumin-coated gold nanoparticles (Cur@AuNPs) and investigated their stability, cytotoxicity, leishmanicidal activity in in vitro and in in vivo experiments. Cur@AuNPs synthesized through a simple one-pot green chemistry technique. The in vitro leishmanicidal activity of curcumin-coated gold nanoparticles against extracellular promastigotes and intracellular amastigotes of protozoan parasite Leishmania major (L. major) was determined by applying the tetrazolium reduction colorimetric quantitative MTT technique. For in vivo assessment, the footpad lesion size and parasite burden in two infection site organs including lymph nodes and footpads of susceptible BALB/c mice infected with L. major were measured. Mice immune responses in all study groups were quantified by measuring the levels of gamma interferon (IFN-γ) and interleukin-4 (IL-4). Viability of Leishmania promastigotes significantly diminished with the inhibition in promastigotes growth (IC50) of 64.79μg/mL and 29.89μg/mL for 24h and 48h, respectively. In vitro nanoparticles treatment efficiently cleared the L. major amastigotes explanted in macrophages but had no harmful toxicity on the mice cells. In the in vivo condition, in the treated infected BALB/c mice the CL lesion size, Leishmania parasite burden, and IL-4 were decreased, while IFN-γ was significantly increased. The results suggest that Cur@AuNP was an effective compound against Leishmania parasite in vitro and in vivo, efficiently induced T-helper 1 (Th1) responses and augmented host cellular immune responses, and ending in a reduced Leishmania parasite burden. Therefore, it may be identified as a novel potential therapeutic approach for the local therapy of zoonotic CL treatment with high cure rates.