Identifying and validating MMP family members (MMP2, MMP9, MMP12, and MMP16) as therapeutic targets and biomarkers in kidney renal clear cell carcinoma (KIRC).

Kidney Renal Clear Cell Carcinoma (KIRC) is a malignant tumor that carries a substantial risk of morbidity and mortality. The MMP family assumes a crucial role in tumor invasion and metastasis. This study aimed to uncover the mechanistic relevance of the MMP gene family as a therapeutic target and diagnostic biomarker in Kidney Renal Clear Cell Carcinoma (KIRC) through a comprehensive approach encompassing both computational and molecular analyses. STRING, Cytoscape, UALCAN, GEPIA, OncoDB, HPA, cBioPortal, GSEA, TIMER, ENCORI, DrugBank, targeted bisulfite sequencing (bisulfite-seq), conventional PCR, Sanger sequencing, and RT-qPCR based analyses were used in the present study to analyze MMP gene family members to accurately determine a few hub genes that can be utilized as both therapeutic targets and diagnostic biomarkers for KIRC. By performing STRING and Cytohubba analyses of the 24 MMP gene family members, MMP2 (matrix metallopeptidase 2), MMP9 (matrix metallopeptidase 9), MMP12 (matrix metallopeptidase 12), and MMP16 (matrix metallopeptidase 16) genes were denoted as hub genes having highest degree scores. After analyzing MMP2, MMP9, MMP12, and MMP16 via various TCGA databases and RT-qPCR technique across clinical samples and KIRC cell lines, interestingly, all these hub genes were found significantly overexpressed at mRNA and protein levels in KIRC samples relative to controls. The notable effect of the up-regulated MMP2, MMP9, MMP12, and MMP16 was also documented on the overall survival (OS) of the KIRC patients. Moreover, targeted bisulfite-sequencing (bisulfite-seq) analysis revealed that promoter hypomethylation pattern was associated with up-regulation of hub genes (MMP2, MMP9, MMP12, and MMP16). In addition to this, hub genes were involved in various diverse oncogenic pathways. The MMP gene family members (MMP2, MMP9, MMP12, and MMP16) may serve as therapeutic targets and prognostic biomarkers in KIRC.

The Effectiveness of Silver Nanoparticles Mixed with Calcium Hydroxide against Candida albicans: An Ex Vivo Analysis.

INTRODUCTION: The purpose of this study was to assess the antifungal activity of silver nanoparticles (AgNPs) in combination with calcium hydroxide (Ca(OH)2) against Candida albicans (C. albicans). METHODS: AgNPs was mixed with pure Ca(OH)2 powder in an aqueous base. A standard suspension (1 × 108 bacterial cells/mL) of C. albicans was prepared in a 96-well plate and incubated on shaker at 37 °C in 100% humidity to allow fungal biofilm formation in infected dentin slices (n = 98). The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of AgNPs alone or with Ca(OH)2 were determined. The samples were separately placed in 24-well tissue culture plates and divided into three experimental groups (0.03, 0.04, and 0.06) and three control groups; negative (saline) and positive chlorhexidine gel and Ca(OH)2. Quantitative measurements of fungal activity by XTT colorimetric assay and qualitative measurements using confocal laser microscopy and scanning electron microscopy were performed. RESULTS: The cell viability of C. albicans in the experimental groups was significantly reduced compared to the negative control group. The combination of (AgNPs (0.04%) and Ca(OH)2) was the most potent against C. albicans. CONCLUSIONS: The findings demonstrated that combining silver nanoparticles with Ca(OH)2 was more effective against C. albicans biofilm compared to Ca(OH)2 alone, suggesting a combing effect.

Antibiofilm Efficacy of Silver Nanoparticles Alone or Mixed with Calcium Hydroxide as Intracanal Medicaments: An Ex-Vivo Analysis.

INTRODUCTION: This study aimed to determine the most effective antibiofilm concentration of silver nanoparticles (AgNPs) alone or in combination with calcium hydroxide (Ca[OH]2) against Fusobacterium nucleatum biofilm. METHODS: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AgNPs alone or with Ca(OH)2 were determined. Dentin slices were sterilized and inoculated with F. nucleatum for 3 weeks to establish a biofilm. Samples were randomly assigned to determine the MIC and MBC for AgNPs alone or mixed with Ca(OH)2. A higher concentration of AgNPs for both preparations was also used. Triple antibiotic paste, Ca(OH)2, and saline were used as controls. Specimens in each group were subdivided over 2 observation periods: 7 and 14 days. At the end of each period, specimens were analyzed with 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to determine the metabolic activity. Also, samples from each group were assessed by scanning electron microscopy. RESULTS: The MIC and MBC of AgNPs alone against F. nucleatum coincided at 0.04%. The combination of AgNPs + Ca(OH)2 exhibited a lower MIC and MBC of 0.03%. MTT analysis showed a significant reduction in bacterial viability in all groups compared with negative controls (P < .05). A more substantial reduction in bacterial cells was observed with increasing concentrations of AgNPs at both periods. The combination (AgNPs [0.06%] + Ca[OH]2) was the most potent against F. nucleatum. CONCLUSIONS: The findings demonstrated that combining AgNPs with Ca(OH)2 was more effective on the F. nucleatum biofilm than either material alone, suggesting a combined effect.

Evaluation of wound healing activity of henna, pomegranate and myrrh herbal ointment blend.

This study assessed the wound healing potential and antimicrobial activity of henna, pomegranate and myrrh extract formulations and their blend in excision, and dead space wound models in rats in comparison to a marketed ointment (gentamycin). The natural extracts were used in ointment formulations alone or in a combination of three extracts at a total concentration of 15% w/w in medications. The percent of wound contraction in case of henna, myrrh, pomegranate, the blend and gentamycin (10 mg/kg) were 85.90-98.5%, 88.35-99.52%, 93.55-100%, 97.30-100%, and 90.25-100% from days 16 to 20, respectively. The blended formulation showed the highest increase in the percent of wound contraction and decrease in the epithelisation period compared to other formulations and showed comparable results to the standard ointment. The histological studies of excision biopsy at day 24 showed healed skin structures with normal epithelisation, the restoration of adnexa and fibrosis within the dermis in all of the formulation- and gentamycin-treated groups while the control group lagged behind in the formation of the amount of ground substance in the granulation tissue. The formulations showed antimicrobial activity against Candida, Staphylococcus aureus, mucous membrane infections and E. coli topical infections. The study proved the wound healing potential and antimicrobial activity of the herbal extract.

Nimesulide/methyl ß-cyclodextrin inclusion complexes: physicochemical characterization, solubility, dissolution, and biological studies.

Nimesulide (NIM) is an insoluble nonsteroidal anti-inflammatory drug (NSAID). Complexation of drug with methyl ß-cyclodextrin was evaluated to improve solubility and dissolution rate of NIM. Complexation was achieved via a coevaporation technique to obtain different drug to polymer molar ratios (1:1, 1:2, and 1:3). The physicochemical characterization of the systems using powder X-ray diffraction and infrared spectroscopy was carried out to understand the influence of this technological process on the physical status of single components and complex systems and to detect possible interactions between drug and carrier. Moreover, quantitative solubility and in vitro dissolution studies of NIM alone and NIM inclusion complexes were studied in the dissolution media of phosphate buffer pH 5.5 and 7.4. The analysis provided existence of a molecular interaction between drug and carrier together in the complex state. The study showed that the inclusion systems enhanced of drug solubility, dissolution rate, and anti-inflammatory activity.

Emulsification/internal gelation as a method for preparation of diclofenac sodium-sodium alginate microparticles.

Emulsification/internal gelation has been suggested as an alternative to extrusion/external gelation in the encapsulation of several compounds including non-steroidal anti-inflammatory drugs such as diclofenac sodium. The objective of the present study was a trial to formulate diclofenac sodium as controlled release microparticles that might be administered once or twice daily. This could be achieved via emulsification/internal gelation technique applying Box-Behnken design to choose these formulae. Box-Behnken design determined fifteen formulae containing specified amounts of the independent variables, which included stirring speed in rpm (X1), drug:polymer ratio (X2) and the surfactant span 80% (X3). The dependent variables studied were cumulative percent release after two hours (Y1), four hours (Y2) and eight hours (Y3). The prepared microparticles were characterized for their production yield, sizes, shapes and morphology, entrapment efficiency and Diclofenac sodium in vitro release as well. The results showed that the production yield of the prepared diclofenac sodium microparticles was found to be between 79.55% and 97.41%. The formulated microparticles exhibited acceptable drug content values that lie in the range 66.20-96.36%. Also, the data obtained revealed that increasing the mixing speed (X1) generally resulted in decreased microparticle size. In addition, scanning electron microscope images of the microparticles illustrated that the formula contains lower span concentration (1%) in combination with lower stirring speed (200 rpm) which showed wrinkled, but smooth surfaces. However, by increasing surfactant concentration, microspheres' surfaces become smoother and slightly porous. Kinetic treatment of the in vitro release from drug-loaded microparticles indicated that the zero order is the drug release mechanism for the most formulae.

Preparation, physicochemical characterization and biological evaluation of cefodizime metal ion complexes.

OBJECTIVES: Cefodizime is a broad spectrum cephalosporin belonging to the third generation agents. In this study, attention has been paid to the preparation, physicochemical characterization and biological evaluation of new Cu2+, Zn2+, Fe3+, Co2+ and Al3+ complexes of cefodizime. METHODS: The stoichiometrics and the mode of bonding of the complexes were deduced from their elemental and metal analysis, electrical conductivity measurements, UV-vis, infrared and Raman spectroscopic investigations. Study of the stoichiometry of these complexes referred to the formation of 1 : 1 ratios of metal to ligand. Antimicrobial activity of the complexes was determined using two strains of Gram-positive (Bacillus subtilis and Proteus vulgaris) and two strains of Gram-negative (Escherichia coli W3110 and Pseudomonas putida) bacteria. The minimal inhibitory concentration was determined as the lowest concentration inhibiting bacterial growth on solid Luria Bertani medium. KEY FINDINGS: The spectra gave evidence as to the position of binding. In addition, the aqueous solubility of cefodizime was strongly reduced by complexation. CONCLUSIONS: The antibacterial activity of cefodizime was not affected by complexation with Al3+ but it was reduced by complexation with the other tested metal ions against the bacteria under study.

Characterization of the interaction of cefadroxil with different metal ions using CE.

The present study describes the application of CE to investigate the interaction between a cephalosporin, cefadroxil and different metal ions. This study aims at quantifying the interaction of cefadroxil with Zn(2+), Al(3+), Fe(3+), Cu(2+) and Co(2+) ions using the effective electrophoretic mobility as a parameter for calculation of the reaction association constant (K). For this purpose, the electrophoretic mobilities of cefadroxil at different metal ion concentrations in citrate buffer pH 3.6 were determined. A mathematical model to calculate the association constant between cefadroxil and metal ions was used. The results showed that a strong change in the cefadroxil electrophoretic mobility was observed by increasing the metal ion concentrations. The results revealed that by increasing the concentration of metal ions in the running buffer the degree of complexation increases too. On the basis of the results, the strength of the interaction of cefadroxil with the investigated metal ions follows the order Zn(2+)>Cu(2+)>Co(2+)>Fe(3+)>Al(3+). The association constant of the investigated reactions ranged from 451.57 to 1546.52 L/mmol. The results indicate that it is possible to characterize the interaction between cefadroxil and metal ions quantitatively using CE.

Effect of Different Metal Ions on the Biological Properties of Cefadroxil.

The effect of different metal ions on the intestinal transport and the antibacterial activity of cefadroxil [(6R,7R)-7-{[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino}-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid] was investigated. The [14C]Gly-Sar uptake via PEPT1 was inhibited by Zn2+ and Cu2+ treatment in a concentration-dependent manner (Ki values 107 ± 23 and 19 ± 5 µM, respectively). Kinetic analysis showed that the Kt of Gly-Sar uptake was increased 2-fold in the presence of zinc sulphate (150 µM) whereas the Vmax value were not affected suggesting that zinc ions inhibited Gly-Sar uptake by PEPT1 in a competitively manner. Ni2+ exhibited moderate inhibitory effect, whereas Co2+, Mg2+, Al3+ ions showed no inhibitory effect on Gly-Sar uptake via PEPT1. Subsequently, we examined the effect of Zn2+ and Al3+ ions on the transepithelial transport of cefadroxil across Caco-2 cells cultured on permeable supports. The results showed that zinc ions inhibited the transepithelial flux of cefadroxil at Caco-2 cell monolayers while Al3+ ions had no effect. The interaction of cephalosporins with the metal ions could suggest negative effects of some metal ions on the clinical aspects of small intestinal peptide and drug transport. Finally, the effect of Zn2+, Cu2+ and Al3+ ions on the antibacterial activity of cefadroxil was tested. It was found that there is no significant difference between the activity of cefadroxil and the cefadroxil metal ion complexes studied against the investigated sensitive bacterial species.