Caspase-Dependent Apoptosis Induced by Simarouba Glauca on Human Non-Small-Cell Lung Cancer, A549 Cells.

BACKGROUND: The leaves of Simarouba glauca (S. glauca) have been used as a potential source of anticancer agents in traditional medicine. Attempts have been made to isolate anticancer agents from the leaves of S. glauca. The objective of the present study was to demonstrate the anticancer and apoptotic effect of the leaf extract of petroleum ether (LPE) on human non-small-cell lung cancer A549 cells. METHODS: MTT assay was used to investigate the effect of LPE on the viability of A-549 cells. The apoptotic effect of human lung cancer cells was evaluated using fluorescence staining, acridine orange/ ethidium bromide staining, Hoechst staining, flow cytometry analysis, annexin V staining, and caspase assay. RESULTS: The results showed a direct correlation between the dose and the rate of cytotoxicity. Fluorescence staining revealed apoptotic features, such as blebbing and chromatin condensation. Flow cytometry analysis and annexin V staining revealed phosphatidyl serine externalization. Caspase assay confirmed that the extract inhibited cell death. Caspase 3 expressions indicated that the cell death occurred either through the mitochondrial pathway or the death receptor. CONCLUSIONS: The study revealed that the LPE induced the apoptosis of human non-small-cell lung cancer, A549 cells, either through mitochondrial or death receptor pathway.

The Apoptotic Properties of Leaf Extracts of Simarouba glauca against Human Leukemic Cancer Cells.

BACKGROUND AND OBJECTIVE: Simarouba glauca is a plant belonging to the family of Simaroubaceae. It is a potent source of secondary metabolites. The aim of this study was to evaluate the apoptotic properties of leaf extracts of Simarouba glauca against human leukemic cancer cells. MATERIALS AND METHODS: Cytotoxicity of Simarouba glauca was assessed in the leaf extract of petroleum ether against leukemic cells by MTT assay. To detect the apoptotic features, fluorescence microscopy analysis was done with dual acridine orange/ethidium bromide fluorescent staining and Hoechst staining. To determine the externalization of phosphatidylserine, annexin v staining was done. Mitochondrial or death receptor activation was confirmed by caspase 3 analysis by flow cytometry. RESULTS: This study revealed that Simarouba glauca was able to treat leukemia. Among the four extracts, petroleum ether extract showed a higher order of in vitro anticancer activity. The petroleum ether extract strongly inhibited the proliferation of K562 cell lines with IC50 values of 186 µg/ml. Dual acridine orange/ethidium bromide fluorescent staining and Hoechst staining revealed the characteristic features of apoptosis. Annexin V confirmed early and late stage apoptosis. Caspase-3 analysis revealed that cell death was due to mitochondrial or death receptor activation in mitochondrial pathway. CONCLUSION: These findings suggested that Simarouba glauca leaf extracts inhibited leukemic cells in a time- and dose-dependent manner either through mitochondrial or death receptor activation. The leaf extracts of Simarouba glauca was found to be nontoxic to lymphocytes. It can be concluded that Simarouba glauca is an important source of phytochemicals posing efficacy against leukemic cancer cells.
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Cytotoxicity Profiling of Annona Squamosa in Cancer Cell Lines.

Objective: In the study our aim was to evaluate the cytotoxic activity of different solvent extracts of Annona squamosa seeds. Methods and materials: The four extracts used were petroleum ether, chloroform, ethyl acetate and methanol were tested using cytotoxicity assays. Results: Among the four extracts tested petroleum ether showed maximum cytotoxicity against a panel of cancer cell lines such as nasopharyngeal cancer (KB) cells, lung cancer (A-549) cells, breast cancer (MCF- 7) cells, leukemic (K-562) cells and inhibited the growth of murine cancer cells such as Dalton's lymphoma ascites (DLA) and Ehrlich ascites carcinoma (EAC). Conclusion: Petroleum ether extract of Annona squamosa seeds showed cytotoxicity towards a panel of cancer cells meanwhile non-significant cytotoxicity towards normal cells.

Occurrence of vancomycin-resistant Staphylococcus aureus in the oral cavity of patients with dental caries.

Oral streptococci are the major group of microbes isolated from oral microflora. They represent frequent pathogens of infective endocarditis (IE), and it is assumed that in most of the cases oral streptococci are acquired via mucosa layer of oral cavity. Staphylococcus aureus is also frequently isolated from IE as it accounts for 20%-30% of all cases. Vancomycin has been the most reliable therapeutic agent against infections caused by methicillin-resistant S. aureus (MRSA). The main objective of this study was to examine the occurrence of S. aureus species in dental caries specimens. Antimicrobial susceptibility testing of S. aureus to four antibiotics namely vancomycin, linezolid, teicoplanin, and daptomycin was performed. Detection of vancomycin resistance was conducted using polymerase chain reaction. Among the tested 150 strains, 98 were MRSA and of that 54 were vancomycin sensitive and 27 were resistant. All 98 MRSA strains were positive for mecA and 36 yielded pvl, whereas 13 carried vanA and only 2 were positive for vanB. Majority of the isolates showed sensitivity toward daptomycin and linezolid. Strains of S. aureus exhibiting decreased susceptibility to different antibiotics like vancomycin, daptomycin, and linezolid severely compromise the therapeutic alternatives and require a considerable amount of time, public awareness, and integrative health-care strategies to prevent the emergence of resistance to these compounds.

Chemical Plaque Control Strategies in the Prevention of Biofilm-associated Oral Diseases.

Dental plaque is a biofilm that forms naturally on the surfaces of exposed teeth and other areas of the oral cavity. It is the primary etiological factor for the most frequently occurring oral diseases, such as dental caries and periodontal diseases. Specific, nonspecific, and ecologic plaque hypothesis explains the causation of dental and associated diseases. Adequate control of biofilm accumulation on teeth has been the cornerstone of prevention of periodontitis and dental caries. Mechanical plaque control is the mainstay for prevention of oral diseases, but it requires patient cooperation and motivation; therefore, chemical plaque control agents act as useful adjuvants for achieving the desired results. Hence, it is imperative for the clinicians to update their knowledge in chemical antiplaque agents and other developments for the effective management of plaque biofilm-associated diseases. This article explores the critical analysis of various chemical plaque control strategies and the current trends in the control and prevention of dental plaque biofilm.

Preparation and characterization of chitosan-natural nano hydroxyapatite-fucoidan nanocomposites for bone tissue engineering.

Solid three dimensional (3D) composite scaffolds for bone tissue engineering were prepared using the freeze-drying method. The scaffolds were composed of chitosan, natural nano-hydroxyapatite (nHA) and fucoidan in the following combinations: chitosan, chitosan-fucoidan, chitosan-nHA, and chitosan-nHA-fucoidan. Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), X-ray diffraction analysis (XRD), scanning electron microscopy (SEM), and optical microscopy (OM) were used to determine the physiochemical constituents and the morphology of the scaffolds. The addition of nHA into the chitosan-fucoidan composite scaffold reduced the water uptake and water retention. FT-IR analysis confirmed the presence of a phosphate group in the chitosan-nHA-fucoidan scaffold. This group is present because of the presence of nHA (isolated via alkaline hydrolysis from salmon fish bones). Microscopic results indicated that the dispersion of nHA and fucoidan in the chitosan matrix was uniform with a pore size of 10-400µm. The composite demonstrated a suitable micro architecture for cell growth and nutrient supplementation. This compatibility was further elucidated in vitro using periosteum-derived mesenchymal stem cells (PMSCs). The cells demonstrated high biocompatibility and excellent mineralization for the chitosan-nHA-fucoidan scaffold. We believe that a chitosan-nHA-fucoidan composite is a promising biomaterial for the scaffold that can be used for bone tissue regeneration.

Multifunctional nano manganese ferrite ferrofluid for efficient theranostic application.

Ferrofluid-based manganese (Mn(2+)) substituted superparamagnetic iron oxide nanoparticles stabilized by surface coating with trisodium citrate (MnIOTCs) were synthesized for enhanced hyperthermic activity and use as negative magnetic resonance imaging (MRI) contrast media intended for applications in theranostics. The synthesized MnIOTC materials were characterized based on their physicochemical and biological features. The crystal size and the particle size at the nano level were studied using XRD and TEM. The presence of citrate molecules on the crystal surface of the iron oxide was established by FTIR, TGA, DLS and zeta potential measurements. The superparamagnetic property of MnIOTCs was measured using a vibrating sample magnetometer. Superparamagnetic iron oxide substituted with Mn(2+) with a 3:1 molar concentration of Mn(2+) to Fe(2+) and surface modified with trisodium citrate (MnIO75TC) that exhibited a high T2 relaxivity of 184.6mM(-1)s(-1) and showed excellent signal intensity variation in vitro. Hyperthermia via application of an alternating magnetic field to MnIO75TC in a HeLa cell population induced apoptosis, which was further confirmed by FACS and cLSM observations. The morphological features of the cells were highly disrupted after the hyperthermia experiment, as evidenced from E-SEM images. Biocompatibility evaluation was performed using an alamar blue assay and hemolysis studies, and the results indicated good cytocompatibility and hemocompatibility for the synthesized particles. In the current study, the potential of MnIO75TC as a negative MRI contrast agent and a hyperthermia agent was demonstrated to confirm its utility in the burgeoning field of theranostics.

A comparative study of the debridement efficacy and apical extrusion of dynamic and passive root canal irrigation systems.

BACKGROUND: Root canal irrigation carries a risk of extrusion of irrigant into the periapical tissues which can be associated with pain, swelling, and tissue damage. Studies have shown less extrusion with sonic or apical negative pressure devices compared with syringe and side-port needle or passive ultrasonic irrigation with continuous irrigant flow. This study aimed to evaluate the effectiveness of the EndoVac irrigation system, regarding 1) debris removal and 2) the control of apically extruded irrigating solution. METHODS: Fifty extracted human single-rooted teeth were used in this study. The teeth were then randomly divided into three experimental groups according to the type of irrigation used and one control group. In group 1, irrigation was performed using the EndoVac irrigation system. In group 2, irrigation was performed using a 30-gauge, tip-vented irrigation needle. In group 3, irrigation was performed using a 30-gauge, side-vented irrigation needle. The control group received instrumentation with no irrigation to serve as a control for cleaning efficiency. Root canal instrumentation was performed using the Profile NiTi rotary system with a crown-down technique. All of the experimental teeth were irrigated with the same amount of 5.25% sodium hypochlorite. The amount of extruded irrigating solution was then measured by subtracting the post-instrumentation weight from the pre-instrumentation weight using an electronic balance. The cleanliness of debris removal was evaluated using scanning electron microscopy. RESULTS: EndoVac irrigation had the least amount of extrusion followed by the side-vented and tip-vented method. The difference between the groups was statistically significant (P <0.01). As for the cleaning results, the debris collection in the EndoVac and tip-vented groups was the least in the apical third. In the control and the side-vented groups, the debris was the greatest in the apical third, but this difference was not significant among the three experimental groups. CONCLUSIONS: The EndoVac irrigation system extruded significantly less irrigant solution than either needle irrigation system. Debris collection was the least in the apical third for the EndoVac irrigation system. No significant difference was found in the cleaning efficiency among the three irrigation systems.

Resolution, sensitivity, and in vivo application of high-resolution computed tomography for titanium-coated polymethyl methacrylate (PMMA) dental implants.

OBJECTIVES: The aims of this study were (i) to determine the spatial resolution and sensitivity of micro- versus nano-computed tomography (CT) techniques and (ii) to validate micro- versus nano-CT in a dog dental implant model, comparative to histological analysis. MATERIAL AND METHODS: To determine spatial resolution and sensitivity, standardized reference samples containing standardized nano- and microspheres were prepared in polymer and ceramic matrices. Thereafter, 10 titanium-coated polymer dental implants (3.2 mm in Ø by 4 mm in length) were placed in the mandible of Beagle dogs. Both micro- and nano-CT, as well as histological analyses, were performed. RESULTS: The reference samples confirmed the high resolution of the nano-CT system, which was capable of revealing sub-micron structures embedded in radiodense matrices. The dog implantation study and subsequent statistical analysis showed equal values for bone area and bone-implant contact measurements between micro-CT and histology. However, because of the limited sample size and field of view, nano-CT was not rendering reliable data representative of the entire bone-implant specimen. CONCLUSIONS: Micro-CT analysis is an efficient tool to quantitate bone healing parameters at the bone-implant interface, especially when using titanium-coated PMMA implants. Nano-CT is not suitable for such quantification, but reveals complementary morphological information rivaling histology, yet with the advantage of a 3D visualization.

Plasma cell gingivitis among herbal toothpaste users: a report of three cases.

AIM: The aim of this article is to present a brief review of plasma cell gingivitis (PCG) along with reports of three cases with varying clinical presentations of the condition associated with the use of herbal toothpaste. BACKGROUND: PCG is a rare benign condition of the gingiva characterized by sharply demarcated erythematous and edematous gingivitis often extending to the mucogingival junction. This is considered a hypersensitive reaction. The histological appearance consists of a dense infiltration of normal plasma cells separated by collagenous stroma, usually confined to the free and attached gingiva. The lesion can be eliminated by identifying and avoiding the source of the allergen. REPORT: Three patients ages 26, 27, and 36, respectively, presented with acutely inflamed gingival and a history of recently switching to herbal toothpaste. The gingiva bled readily on probing. Blood tests and gingival biopsy were not contributory. Patients were advised to refrain from the use of herbal toothpaste, and, along with periodontal treatment, the condition underwent remission within a week to two weeks in all three cases. SUMMARY: As more and more herbal products are gaining popularity, clinicians should be aware of some of the untoward effects of these products. Since PCG mimics lesions associated with leukemia and myeloma an early diagnosis of the condition is vital.