Exploring the in vitro potential of royal jelly against glioblastoma and neuroblastoma: impact on cell proliferation, apoptosis, cell cycle, and the biomolecular content.

Neuroblastoma and glioblastoma are the most commonly seen nervous system tumors, and their treatment is challenging. Relatively safe and easy acquisition of nutraceutical natural products make them suitable candidates for anticancer research. Royal jelly (RJ), a superfood, has many biological and pharmacological activities. This study was conducted to, for the first time, elucidate its anticancer efficiency, even in high doses, on neuroblastoma and glioblastoma cell lines through cell viability, apoptosis, cell cycle and biomolecular content evaluation. We performed experiments with RJ concentrations in the range of 1.25-10 mg mL-1 for 48 h. Cell viability assays revealed a notable cytotoxic effect of RJ in a concentration-dependent manner. Treatment with a high dose of RJ significantly increased the apoptotic cell population of both cell lines. Furthermore, we observed G0-G1 phase arrest in neuroblastoma cells but G2-M arrest in glioblastoma cells. All these cellular changes are closely associated with the alterations of the macromolecular makeup of the cells, such as decreased saturated lipid, protein, DNA and RNA amounts, protein conformational changes, decreased protein phosphorylation and increased protein carbonylation. These cellular changes are associated with RJ triggered-ROS formation. The clear segregation between the control and the RJ-treated groups proved these changes, obtained from the unsupervised and supervised chemometric analysis. RJ has good anticancer activity against nervous system cancers and could be safely used with current treatment strategies.

Evaluation of cytotoxic and apoptotic effects of the extracts and phenolic compounds of Astragalus globosus Vahl and Astragalus breviflorus DC.

Astragalus L. is a genus member of the Fabaceae family, representing about 3,000 species all over the world and 380 species in Turkey. Astragalus species have been used in traditional medicine for many years. Astragalus globosus Vahl, known as "top geven", is a dwarf, scapose, perennial herb, Astragalus breviflorus DC., known as "yünlü geven", is an extremely spiny dwarf shrub. These endemic species grow in the Turkish cities of Erzurum, Kars, and Van. This is the first phytochemical and cytotoxic investigation of Astragalus globosus Vahl and Astragalus breviflorus DC. The main extracts and sub-fractions from the plants were evaluated for in vitro cytotoxic and apoptotic activities. The IC50 values of dichloromethane, n-butanol, and water extracts of the aerial parts of A. globosus against the MCF-7 cell line were determined as 28.39, 868.60, and 1753.00 µg/mL. The values for the MDA-MB-231 cell line were 264.00, 620.30, and 1300.50 µg/mL, respectively. From A. globosus, the following were isolated: a flavone glycoside, diosmetin-7-O-rutinoside (1); and two flavonol glycosides, isorhamnetin-3-O-rutinoside (2) and quercetin-3-O-galactoside (3). From A. breviflorus, two phenolic acids, caffeic acid (4) and chlorogenic acid (5), and a flavan-3-ol, catechin (6), were isolated. Diosmetin-7-O-rutinoside was isolated from Astragalus species for the first time and showed the highest cytotoxic activities on the MCF-7 and MDA-MB-231 breast cancer cell lines with IC50 values of 13.65 and 12.89 µg/mL, respectively. Moreover, we observed that diosmin exerts cytotoxic effects by causing cell necrosis.

Preparation and Characterization of Nanosuspensions of Triiodoaniline Derivative New Contrast Agent, and Investigation into Its Cytotoxicity and Contrast Properties.

Iodine-based contrast agents have limitations such as rapid clearance, potential renal toxicity, non-specific blood pool distribution, headache, and adverse events. Nowadays, it is quite common to work with nanosized systems in order to eliminate the side effects of contrast agents. This study aims to synthesize a new iodinated contrast agent, prepare its nanosuspension by using the nanoprecipitation method, investigate its cytotoxicity, and compare its contrast properties with iohexol and iopromide through in-vitro experiments. The values of nanosuspension particle size and zeta potential have been found to be ~ 400 nm and ~ (-) 15 mV, respectively. In-vitro cellular viability findings indicated that the nanosuspension has lower cytotoxicity than the iohexol and iopromide. In the computed tomography (CT) imaging study of contrast features of nanosuspensions and two commercial agents, which involved 86 CT examinations using 31 parameters and two different devices, it was found that iodine had a stronger presence in its nanosuspension form than in iohexol and iopromide, which were the other two commercial contrast agents, when used in equal amounts. Thus in the case of nanosuspensions contrast brightness was achieved by using less iodine, while the same brightness could be obtained with higher doses of iohexol and iopromide. CT imaging therefore be done without much chemical use, which indicates that it may witness fewer side effects in the future.

The cytotoxic and apoptotic effects of beta-blockers with different selectivity on cancerous and healthy lung cell lines.

ß-blockers having specific affinities to ß-adrenergic receptors are routinely used to treat cardiovascular problems. Additionally, it has been demonstrated that these drugs can be effective in treating apoptosis-related diseases. The current study was conducted to investigate the cytotoxic and apoptotic effects of ß-1 selective esmolol, ß-2 selective ICI-118,551, and non-selective nadolol blockers on the cancerous and healthy lung cells. MTT test was used to evaluate cytotoxicity. Apoptotic actions were examined by using Annexin V-FITC/PI assay, JC-1 staining, ROS test, and the determination of the caspase-4 and -9, Bcl-2, Bax, Bax/Bcl-2, and JNK levels. Although the MRC-5 showed greater resistance than A549 cells, the ß-blockers at 150-250 µM exhibited different levels of cytotoxic effect on both lung cell lines. Esmolol was found to be the most ineffective blocker and the increases in Bcl-2 protein levels were appeared to be effective in resistance to this drug. The increases in reactive oxygen species (ROS) together with the increase in caspase-4 and Bax protein levels have been shown to play a role in ICI-118,551 induced lung cell death. Nadolol was the most effective blocker increasing the total apoptotic cell population in both lung cells, which was based on both mitochondrial and endoplasmic reticulum stress. When the selectivities of the ß-blockers are considered, it seems that ß-2 specific antagonism predominantly mediated the death of lung cells, and the overwhelming factors causing apoptosis mainly varied depending on the selectivity of the blockers.

Combining vancomycin-modified gold nanorod arrays and colloidal nanoparticles as a sandwich model for the discrimination of Gram-positive bacteria and their detection via surface-enhanced Raman spectroscopy (SERS).

This study reports the development of a highly sensitive antibiotic-based discrimination and sensor platform for the detection of Gram-positive bacteria through surface-enhanced Raman spectroscopy (SERS). Herein, a combination of gold nanorod arrays (GNAs) and colloidal gold nanoparticles (AuNPs) was used as a SERS platform. To specifically capture Gram-positive bacteria, both GNAs and AuNPs were functionalized with thiol-modified vancomycin (HS-Van) molecules. Three different strains of bacteria (Bacillus subtilis and Staphylococcus aureus as Gram-positive, and Escherichia coli as Gram-negative) were employed to test the proposed system. HS-Van functionalized GNAs (GNA@Van) captured Gram-positive bacteria with high specificity. Also, the bacteria captured by GNA@Van (GNA@Van@Bct) systems showed high signal-to-noise SERS signals with high reproducibility. Addition of AuNP@Van to GNA@Van@Bct resulted in the emergence of a sandwich system (GNA@Van@Bct@Van@AuNP). This system led to a further enhanced SERS signal. The chemometric analysis of GNA@Van@Bct@Van@AuNP enabled the obvious discrimination and detection of Gram-positive bacteria. For comparison, we also tested a smooth gold surface with the same procedure and a similar trend was observed with lower SERS activity.

Niobium-oxynitride coatings for biomedical applications: Its antibacterial effects and in-vitro cytotoxicity.

316L Stainless Steel (SS) has been widely used in many medical applications, such as orthopedic prostheses and cardiovascular implants due to its good mechanical properties and resistance to corrosion. Despite its superior features, SS has bio-functionality problems. In this study, niobium oxynitride coatings were deposited onto 316L SS substrates to improve their biocompatibility using a reactive radio frequency (RF) magnetron sputtering technique. The nitrogen flow was fixed, and the nitrogen to oxygen flow ratio was set to 2, 5 and 10 to investigate the effect of oxygen concentration on biocompatibility and the antibacterial behavior of the oxynitride films. The microstructure, morphology and wettability properties of the coatings were analyzed by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and an optical tensiometer. The antibacterial activities of coated and uncoated 316L substrates were tested against S. aureus and E. coli bacterial strains. The cytotoxic effects of NbOxNy-coated and uncoated substrates were evaluated on human fibroblast cells. The results showed that niobium oxynitride coatings were not cytotoxic and exhibited more antibacterial activity in comparison to the uncoated ones.

Gastroprotective effects of oleuropein and thymol on indomethacin-induced gastric ulcer in Sprague-Dawley rats.

Ethnopharmacological studies demonstrated that thymol (Thym) and oleuropein (Ole) have therapeutic potential for gastric ulcers. The molecular mechanism underlying the gastroprotective effects of these compounds have not been elucidated yet especially for their individual and combination use at high dose. Therefore, this study was conducted to explore their gastroprotective mechanisms on indomethacin (Indo)-induced gastric ulcer model. Ole (50,100, 250, and 500 mg/kg) and Thym (50,100, 200, and 500 mg/kg) were orally administered to the rats 10 min before the induction of ulcer with Indo. The combination of 500 mg/kg doses of Ole and Thym were applied. The gastric mucosa was evaluated histopathologically. Moreover, TAC/TOS, tumor necrosis factor-alpha (TNF-α), prostaglandin E2 (PGE2), endothelial nitric oxide synthase (eNOS), and caspase-3 levels were assessed by ELISA and the caspase-3 and TNF-α expressions were quantified by qRT-PCR. Indo-induced histopathological changes while Ole and Thym pretreatment prevented these effects. Unlike the 500 mg/kg dose of Ole treatment, the 500 mg/kg dose of Thym administration enhanced these damages. The decreased TAC, PGE2 levels and increased TOS, eNOS, TNF-α, caspase-3 levels were obtained in Indo group. However, these changes were reversed by Ole and Thym groups except the 500 mg/kg dose of Thym and the combination treatment groups. Similar trends were observed in the caspase-3 and TNF-α expression levels. These results demonstrated that enhanced inflammation, oxidant/antioxidant imbalance, and apoptotic activities were occurred in Indo, 500 mg/kg dose of Thym and the combination treatment groups while not in the other groups. The findings demonstrated the gastroprotective ability of Ole and low doses of Thym in gastric ulcer models.

A holistic study on potential toxic effects of carboxylated multi-walled carbon nanotubes (MWCNTs-COOH) on zebrafish (Danio rerio) embryos/larvae.

Multi-walled carbon nanotubes (MWCNTs) have widespread use in industrial and consumer products and great potential in biomedical applications. This leads to inevitably their release into the environment and the formation of their toxic effects on organisms. These effects can change depending on their physicochemical characteristics. Therefore, the toxicological findings of MWCNTs are inconsistent. Their toxicities related to surface modification have not been elucidated in a holistic manner. Hence, this study was conducted to clarify their potential toxic effects on zebrafish embryos/larvae in a comprehensive approach using morphologic, biochemical and molecular parameters. Zebrafish embryos were exposed to 5, 10, 20 mg/L doses of MWCNTs-COOH at 4 h after fertilization and grown until 96 hpf. Physiological findings demonstrated that they induced a concentration-dependent increase in the mortality rate, delayed hatching and decrease in the heartbeat rate. Moreover, it caused abnormalities including yolk sac edema, pericardial edema, head, tail malformations, and vertebral deformities. These effects may be due to the alterations in antioxidant and immune system related gene expressions after their entry into zebrafish embryo/larvae. The entry was confirmed from the evaluation of Raman spectra collected from the head, yolk sac, and tail of control and the nanotube treated groups. The gene expression analysis indicated the changes in the expression of oxidative stress (mtf-1, hsp70, and nfkb) and innate immune system (il-1ß, tlr-4, tlr-22, trf, and cebp) related genes, especially an increased in the expression of the hsp70 and il-1ß. These findings proved the developmental toxicities of MWCNTs-COOH on the zebrafish embryos/larvae.

Diagnosis of malignant pleural mesothelioma from pleural fluid by Fourier transform-infrared spectroscopy coupled with chemometrics.

This study was conducted to differentiate malignant pleural mesothelioma (MPM) from lung cancer (LC) and benign pleural effusion (BPE) from pleural fluids using the diagnostic power of Fourier transform-infrared spectroscopy with attenuated total reflectance mode coupled with chemometrics. Infrared spectra of MPM (n = 24), LC (n = 20), and BPE (n = 25) were collected, and hierarchical cluster analysis (HCA) and principal component analysis (PCA) were applied to their spectra. HCA results indicated that MPM was differentiated from LC with 100% sensitivity and 100% specificity and from BPE, with 100% sensitivity and 88% specificity, which were also confirmed by PCA score plots. PCA loading plots indicated that these separations originated mainly from lipids, proteins, and nucleic acids-related spectral bands. There was significantly higher lipid, protein, nucleic acid, and glucose contents in the MPM and LC. However, the significant changes in triglyceride and cholesterol ester content, protein and nucleic acid structure, a lower membrane fluidity, and higher membrane order were only observed in the MPM. To check the classification success of some test samples/each group, soft independent modeling of class analogies was performed and 96.2% overall classification success was obtained. This approach can provide a rapid and inexpensive methodology for the efficient differentiation of MPM from other pleural effusions.

Quick discrimination of heavy metal resistant bacterial populations using infrared spectroscopy coupled with chemometrics.

Lead and cadmium are frequently encountered heavy metals in industrially polluted areas. Many heavy metal resistant bacterial strains have a high biosorption capacity and thus are good candidates for the removal of toxic metals from the environment. However, as of yet there is no accurate method for discrimination of highly adaptive bacterial strains among the populations present in a given habitat. In this study, we aimed to find distinguishing molecular features of lead and cadmium resistant bacteria using Attenuated Total Reflectance-Fourier Transformed Infrared (ATR-FT-IR) spectroscopy and chemometric approaches. Our results demonstrated that both control and metal exposed E. coli and S. aureus strains could be successfully discriminated from each other using hierarchical cluster and principal component analysis methods. Moreover, we found that lead exposed bacterial strains could be successfully discriminated from cadmium exposed ones with a high heterogeneity value. These clear discriminations can be described by the ability of a bacterium to change its metabolism in terms of the content and structure of cellular macromolecules under heavy metal stress. In our case, cadmium and lead-induced genetic response systems in bacteria caused remarkable alterations in overall cellular metabolism. Bacteria deal with a heavy metal stress by altering nucleic acid methylations and lipid and protein synthesis. Heavy metal burden led to the development of relevant metabolic changes in proteins, lipids, and nucleic acids of the resistant bacteria described in this study. Our approach showed that infrared spectra obtained via ATR-FT-IR spectroscopy coupled with chemometric analysis can be utilized for rapid, low-cost, informative, reliable, and operator-independent discrimination of resistant bacterial populations.

Vitamin A deficiency induces structural and functional alterations in the molecular constituents of the rat hippocampus.

To date, no structural study has been carried out on the effects of vitamin A deficiency (VAD) on hippocampal macromolecules. Therefore, in the present study, the effect of dietary VAD on the structure, content and function of rat hippocampal molecules was investigated using Fourier transform IF spectroscopy. Male Wistar rats were randomly divided into three groups: an experimental group maintained on a vitamin A-deficient liquid diet (VAD, n 7); a control group maintained on a vitamin A-supplemented liquid diet (CON, n 9); a pure control group maintained on standard solid laboratory chow (PC, n 7). The PC group was included in the study to ensure that the usage of liquid diet did not influence the outcomes of VAD. Both the CON and PC groups were successfully discriminated from the VAD group by principal component analysis and hierarchical cluster analysis. The spectral analysis indicated a significant decrease in the contents of saturated and unsaturated lipids, cholesteryl esters, TAG and nucleic acids in the VAD group when compared with the CON group (P≤ 0·05). In addition, a significant decrease in membrane fluidity and a significant increase in lipid order (e.g. acyl chain flexibility) were observed in the VAD group (P≤ 0·001). The results of the artificial neural network analysis revealed a significant decrease in the α-helix structure content and a significant increase in the turn and random coil structure contents, indicating protein denaturation, in the VAD group when compared with the CON and PC groups (P≤ 0·05). Dietary exclusion of vitamin A for 3 months apparently had an adverse impact on compositional, structural and dynamical parameters. These changes can be due to increased oxidative stress, confirming the antioxidant protection provided by vitamin A when used as a dietary supplement at low-to-moderate doses.

Structural and functional characterization of simvastatin-induced myotoxicity in different skeletal muscles.

BACKGROUND: Statins are the most commonly used drugs for the treatment of hypercholesterolemia. Their most frequent side effect is myotoxicity. To date, it remains unclear whether statins preferentially induce myotoxicity in fast- or in slow-twitch muscles. Therefore, we investigated these effects on fast- (extensor digitorum longus; EDL), slow- (soleus; SOL), and mixed-twitch muscles (diaphragm; DIA) in rats by comparing their contractile and molecular structural properties. METHODS: Simvastatin-induced functional changes were determined by muscle contraction measurements, and drug-induced molecular changes were investigated using Fourier transform infrared (FTIR) and attenuated total reflectance (ATR) FTIR spectroscopy. RESULTS: With simvastatin administration (30 days, 50mg/kg), a depression in the force-frequency curves in all muscles was observed, indicating the impairment of muscle contractility; however, the EDL and DIA muscles were affected more severely than the SOL muscle. Spectroscopic findings also showed a decrease in protein, glycogen, nucleic acid, lipid content and an increase in lipid order and lipid dynamics in the simvastatin-treated muscles. The lipid order and dynamics directly affect membrane thickness. Therefore, the kinetics and functions of membrane ion channels were also affected, contributing to the statin-induced impairment of muscle contractility. Furthermore, a reduction in α-helix and ß-sheet and an increase in random coil, aggregated and antiparallel ß-sheet were observed, indicating the protein denaturation. Spectral studies showed that the extent of molecular structural alterations in the muscles following simvastatin administration was in the order EDL>DIA>SOL. CONCLUSIONS: Simvastatin-induced structural and functional alterations are more profound in the fast-twitch than in the slow-twitch muscles. GENERAL SIGNIFICANCE: Myotoxic effects of simvastatin are primarily observed in the fast-twitch muscles.

Low dose simvastatin induces compositional, structural and dynamic changes in rat skeletal extensor digitorum longus muscle tissue.

Statins are commonly used drugs in the treatment of hypercholesterolaemia. There are many adverse effects of statins on skeletal muscle, but the underlying mechanisms remain unclear. In the present study, the effects of low dose (20 mg/kg) simvastatin, a lipophilic statin, on rat EDL muscle (extensor digitorum longus muscle) were investigated at the molecular level using FTIR (Fourier-transform infrared) spectroscopy. FTIR spectroscopy allows us rapid and sensitive determination of functional groups belonging to proteins, lipids, carbohydrates and nucleic acids simultaneously. The results revealed that simvastatin treatment induces a significant decrease in lipid, nucleic acid, protein and glycogen content. A significant increase in the lipid/protein and nucleic acid/protein ratios was also obtained with simvastatin treatment. Furthermore, an increase in lipid order and membrane fluidity was detected. A decrease in the bandwidth of the amide I band and shifting of the position of this band to higher frequency values in treated muscle indicates structural changes in proteins. Detailed secondary structure analysis of the amide I band revealed a significant increase in antiparallel and aggregated beta-sheet, random coil structure and a significant decrease in beta-sheet structure, which indicates protein denaturation.