Analyzing antibiotic residues in honey samples using liquid chromatography-tandem mass spectrometry.

This study aimed to present a sensitive, accurate, and precise analytical method for the determination of 32 antibiotics from 5 groups (sulfonamides, macrolides, aminoglycosides, tetracyclines, and quinolones) and some individual antibiotics (lincomycin, griseofulvin, and 5-hydroxy-flunixin) in 63 honey samples collected from Tehran market. In the presented method, the samples were hydrolyzed by 1% HFBA (hepta fluoro butyric acid) in water, purified on Strata XL polymeric reversed-phase cartridges, and finally analyzed by reversed-phase ion-pair liquid chromatography-electrospray ionization tandem mass spectrometry (RP-IP-LC-ESI-MS/MS). Good performance characteristics were gained for recovery, precision, range, and linearity, the limit of detections (LODs), and the limit of quantifications (LOQs). According to the presented results and considering the absence of permissible limits for antibiotics in honey, 74.6% of the tested samples had antibiotic residues more than the LOQ of the method. The results show that the validated method is suitable for simultaneously detecting antibiotic residues in honey.

Hybrid Analogues of Hydrazone and Phthalimide: Design, Synthesis, In vivo, In vitro, and In silico Evaluation as Analgesic Agents.

BACKGROUND: Based on the anti-inflammatory and analgesic activity of hydrazone and phthalimide, a new series of hybrid hydrazone and phthalimide pharmacophores was prepared and evaluated as analgesic agents. METHODS: The designed ligands were synthesized by reaction of the appropriate aldehydes and 2- aminophthalimide. Analgesic, cyclooxygenase inhibitory, and cytostatic activity of prepared compounds were measured. RESULTS: All the tested ligands demonstrated significant analgesic activity. Moreover, compounds 3i and 3h were the most potent ligands in the formalin and writhing tests, respectively. Compounds 3g, 3j, and 3l were the most COX-2 selective ligands and ligand 3e was the most potent COX inhibitor with a 0.79 of COX-2 selectivity ratio. The presence of electron-withdrawing moieties with hydrogen bonding ability at the meta position was found to affect the selectivity efficiently, in which compounds 3g, 3l, and 3k showed high COX-2 selectivity, and compound 3k was the most potent one. The cytostatic activity of selected ligands demonstrated that compounds 3e, 3f, 3h, 3k, and 3m showed good analgesic and COX inhibitory activity and were less toxic than the reference drug. CONCLUSION: High therapeutic index of these ligands is one of the valuable advantages of these compounds.

Design, synthesis, and biological evaluation of new 2-(4-(methylsulfonyl)phenyl)-N-phenylimidazo[1,2-a]pyridin-3-amine as selective COX-2 inhibitors.

Cyclooxygenase (COX), which plays a role in converting arachidonic acid to inflammatory mediators, could be inhibited by non-steroidal anti-inflammatory drugs (NSAIDs). Although potent NSAIDs are available for the treatment of pain, fever, and inflammation, some side effects, such as gastrointestinal ulcers, limit the use of these medications. In recent years, selective COX-2 inhibitors with a lower incidence of adverse effects attained an important position in medicinal chemistry. In order to introduce some new potent COX-2 inhibitors, a new series of 2-(4-(methylsulfonyl)phenyl)-N-phenylimidazo[1,2-a]pyridin-3-amines was designed, synthesized, and evaluated. The docking studies performed by AutoDock Vina demonstrated that docked molecules were positioned as well as a crystallographic ligand in the COX-2 active site, and SO2Me pharmacophore was inserted into the secondary pocket of COX-2 and formed hydrogen bonds with the active site. The designed compounds were synthesized through two-step reactions. In the first step, different 1-(4-(methylsulfonyl)phenyl)-2-(phenylamino)ethan-1-one derivatives were obtained by the reaction of aniline derivatives and α-bromo-4-(methylsulfonyl)acetophenone. Then, condensation of intermediates with different 2-aminopyridines gave final compounds. Enzyme inhibition assay and formalin test were performed to evaluate the activity of these compounds. Among these compounds, 8-methyl-2-(4-(methylsulfonyl)phenyl)-N-(p-tolyl)imidazo[1,2-a]pyridin-3-amine (5n) exhibited the highest potency (IC50 = 0.07 µM) and selectivity (selectivity index = 508.6) against COX-2 enzyme (selectivity index: COX-1 IC50/COX-2 IC50). The antinociceptive activity assessment via the formalin test showed that nine derivatives (5a, 5d, 5h, 5i, 5k, 5q, 5r, 5s, and 5t) possessed significant activity compared with the control group with a p value less than 0.05.

Design, Synthesis, Docking Studies, Enzyme Inhibitory and Antiplatelet Aggregation Activities of New 1,3-Diphenyl-3-(Phenylthio)Propan-1-One Derivatives as Selective COX-2 Inhibitors.

BACKGROUND: Cancer is the second leading cause of death worldwide after heart disease. A vast number of studies indicated that selective cyclooxygenase-2 (COX-2) inhibitors could be chemopreventive against different types of cancer because the expression of COX-2 is increased. Therefore, to develop new therapeutics for cancer, the design and synthesis of new COX-2 inhibitors with few side effects seem attractive as anti-cancer agents. OBJECTIVE: Some of the well-known drugs that have been widely used for some time have been removed from the market due to the cardiac side effects they cause, so there is a need to introduce a scaffold that can inhibit COX-2 with high potency and low side effects. This study aimed to introduce a new COX-2 inhibitor structure. METHODS: A new series of ß-aryl-ß-mercapto ketones possessing a methylsulfonyl pharmacophore was synthesized and evaluated as selective COX-2 inhibitors. In-vitro COX-1 and COX-2 inhibition effects of these compounds were evaluated, and molecular modeling was examined. Also, the antiplatelet aggregation activity of the synthesized compounds was tested. RESULTS: In-vitro COX-1 and COX-2 inhibition assays indicated that almost all newly synthesized compounds showed selectivity for COX-2 with IC50 values in the 0.07-0.22 µM range and COX-2 selectivity indexes in the 170 to 703.7 range. Among the tested compounds 1-(4-(methylsulfonyl)phenyl)-3-phenyl-3-(phenylthio)propan-1-one (4a), 3-(3,4- dimethoxyphenyl)-1-(4-(methylsulfonyl)phenyl)-3-(phenylthio)propan-1-one (4g) and 3-(4-fluorophenyl)-1-(4-(methyl sulfonyl)phenyl)-3-(phenylthio)propan-1-one (4h) were the most potent COX-2 inhibitors and 3-(3,4- dimethoxyphenyl)-1-(4-(methylsulfonyl)phenyl)-3-(phenylthio)propan-1-one had the highest selectivity index for COX-2 enzyme inhibitory activity. The Anti-platelet aggregation activity results indicated that the compound 1-(4- (methylsulfonyl)phenyl)-3-(phenylthio)-3-(p-tolyl)propan-1-one (4b) possesses the strong anti-platelet activity. Our molecular modeling studies also indicated that the methylsulfonyl pharmacophore group is placed into the adjunct pocket in the COX-2 active site and forms hydrogen bond interactions with NH of Arg513 and NH of His90. CONCLUSION: In brief, all designed and synthesized compounds showed moderate to good COX-2 inhibitory effects and showed good anti-platelet activity. Therefore, these compounds have the potential for further research into developing anti-cancer agents.

Multivariate Optimization and Validation of a Modified QuEChERS Method for Determination of PAHs and PCBs in Grilled Meat by GC-MS.

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are recognized as carcinogens and mutagenic food contaminants that threaten public health. As for food safety aspects, control of these contaminants in processed and fatty food is necessary. In this study, eleven factors were screened by the Plackett-Burman design, and four variables were chosen to optimize with the central composite design (CCD) for the improvement of extraction and cleanup procedures of these food contaminants. The optimized variables include 5 g of sample, 2 mL mixture of 2/2/1 ethyl acetate/acetone/isooctane, 1.6 g of ammonium formate, 0.9 g of sodium chloride, and 0.25 g of sorbent Z-Sep+. A 5 min cleanup vortex time with the spike calibration curve strategy, analyzed by gas chromatography-mass spectrometry (GC-MS), led to the validated limits of quantification (LOQs) for 16 PAHs and 36 PCBs of 0.5-2 and 0.5-1 ng/g, respectively, and recoveries of 72-120%, with an average relative standard deviation (%RSD) of 17, for PAHs, and 80-120%, with an %RSD of 3, for PCBs. The method introduces excellent accuracy, precision, and efficiency, and minimizes matrix effects, and ensures a control procedure, adopted with international standards, for food authorities to determine the contaminants of interest in processed meat, and consequently, prevent food-borne disease to improve public health indices.

Route exposure and adverse effects monitoring of Aflatoxin B1 in the workers of wet waste management, the role of body redox system modulation.

Exposure to dust, containing different fungi metabolites such as aflatoxins is a risk factor for developing liver and kidney health abnormalities. Occupational evaluation of the aflatoxin's exposure-induced health abnormalities should include the monitoring of bioaerosols in the workplace and personal air, and applying of appropriate blood biomarkers to assess Aflatoxin B1 (AFB1) detrimental effects on a worker's health. However, to the best of our knowledge, these appropriate methods, especially determining the associated-adverse effects on health, following exposure, haven't been well documented in the literature at the wet waste handling sites. In the current study, the AFB1 quantity in the area, personal, and settled dust in wet household waste handling samples and AFB1-Albumin levels in the serum of workers in comparison with the control group were determined using high-pressure liquid chromatography with a fluorescent detector (HPLC-FLD) methods. Moreover, the adverse effects of AFB1 on the liver and kidney biochemical profiles of the exposed workers and its relation to antioxidant capacity in the household wet waste sorting were recorded in a consolidated investigation. The results demonstrated that the average airborne dust concentration and its associated AFB1 content were significantly higher in wet waste management sections as compared to the control place, corresponding to the serum AFB1-Albumin levels of workers. Furthermore, AFB1-induced changes in the serum biochemicals evaluating liver and kidney function tests and antioxidant profiles of workers in wet waste handling sections were indicative of their function abnormalities. The results imply AFB1-induced adverse effects on the liver and kidney functions may be mediated through the body redox system modulation.

Synthesis, evaluation of drug delivery potential, and the quantum chemical investigation on a molecular imprinted polymer for quetiapine antipsychotic; a joint experimental and density functional theory study.

In this project, the quetiapine drug was used as the template for synthesis of a molecular imprinted polymer (MIP). The polymerization approach for preparation of this composite was precipitation, where methacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA), and 2,2-azobisissobutyronitrile (AIBN) were used as the functional monomer, the cross-linker, and the initiator, respectively. Scanning electron microscopy (SEM) showed that the diameter of the nanoparticles is about 70 nm. The adsorption rates of quetiapine to the MIP host were evaluated at different pHs, and the results showed that the highest adsorption values were obtained at pH = 7. Moreover, the kinetics of the adsorption process was detected to follow the Langmuir isotherm (R2 = 0.9926) and the pseudo-second-order kinetics (R2 = 0.9937). The results confirmed the high capability of the synthesized MIPs as pharmaceutical carriers for quetiapine. Furthermore, the kinetics of the drug release from the MIP follows the Higuchi model at the pHs of 5.8-6.8 and the Korsmeyer-Peppas model at the pHs of 1.2-5. Finally, in light of the density functional theory (DFT)-based quantum chemical descriptors, the polymer-quetiapine drug complex was designed and investigated. The results showed that there is a strong interaction between the host (polymer) and the guest (drug) due to several hydrogen bonds and other intermolecular (polar) interactions.

A Neglected Challenge in the Analysis of Active Pharmaceutical Substances with Aldehyde Functional Group in Aqueous Matrices.

Aldehydes are compounds that are widely used and popular in organic synthesis due to their high reactivity. This advantage is a disadvantage in medicinal chemistry. Due to the ability of aldehydes to participate in nucleophilic reactions (especially in aqueous biological media) and access to nucleophiles such as amino acids and nucleic acids, drugs with aldehyde functional groups are always used with caution and carefully quantified in biological fluids. Our experience in working on biologically active aldehydes indicates the transformation of these groups of compounds in aqueous or alcoholic solution and thus the failure of analytical methods for their accurate monitoring in such media. Both mass spectrometry and Proton nuclear magnetic resonance spectroscopic findings indicate the reaction of spiramycin with water molecules in an aqueous solution, resulting in the conversion of spiramycin to a new molecule with 18 mass unit difference and thus, the residue amount which is measured and reported based on a mass spectrometries method does not show the correct amount of spiramycin in these samples.

Simultaneous Analysis and Efficient Separation of Anabolic Androgenic Steroids in Dietary Supplement by a Validated HPTLC Method.

Background: Using sports supplements is common among athletes. The presence of anabolic steroids in sports supplements as a hormonal contaminant can increase production efficiency. Since anabolic steroids cause health problems and result in positive doping tests in athletes, it is important to investigate their presence in the supplement preparations consumed by athletes. Objectives: This paper aims to simultaneously determine ten anabolic steroids by high-performance thin-layer chromatography (HPTLC) method in sports supplements. Methods: Chromatographic analysis was conducted on glass silica gel 60F254 plates. The extracts loaded on silica gel plates are subjected to programed multiple development (PMD) to separate anabolic androgenic steroids (AASs). Densitometric scanning is carried out at the wavelength of 245 and 366nm. The method was validated according to the ICH guidelines. Results: Spots at retardation factor (Rf) 0.72 (elution system 1), 0.4 (elution system 1), 0.29 (elution system 2), 0.25 (elution system 2), 0.1 (elution system 1), 0.65 (elution system 2), 0.59 (elution system 1), 0.44 (elution system 1), 0.8 (elution system 3), and 0.82 (elution system 3) values were recognized as 19-nor androstenedione, 19-nortestosterone, methyl testosterone, clostebol, stanozolol, trenbolone enanthate, oxymetholone, oxandrolone, testosterone enanthate, and nandrolone decanoate, respectively. The linear ranges were 25 - 250 µg/mL for oxymetholone, 7 - 50 µg/mL for 19-nor androstenedione, 19-nortestosterone, and oxandrolone, and 3 - 20 µg/mL for methyl testosterone, clostebol, stanozolol, trenbolone enanthate, testosterone enanthate, and nandrolone decanoate. The developed method is validated by acceptable precision (CV < 20%) and good accuracy (94% < R < 114%). The value of limit of detection (LOD) for all derivatives was in the range of 0.02 - 0.16 µg/spot (20-160 µg/g of supplement), while limit of quantitation (LOQ) was found to be in the range of 0.06 - 0.5 µg/spot (60 - 500 µg/g of supplement). Fifty sports supplement samples as real sample were collected and analyzed. None of the samples screened positive using the HPTLC method. Conclusions: In the present study, the fast, cheap, and simple HPTLC method could be used for the multi-residue analysis of ten anabolic androgenic steroids in sports supplements.

Application of the oxycodone templated molecular imprinted polymer in adsorption of the drug from human blood plasma as the real biological environment; a joint experimental and density functional theory study.

In this project, we have synthesized and used a molecular imprinted polymer (MIP) for adsorption of oxycodone residue from the biological samples. Indeed, this study aims to develop a suitable method for determination of oxycodone drug residue in the human plasma using the common analysis methods. Therefore, the MIP was used for the solid phase extraction (MIP-SPE) approach in order to collect the oxycodone opioid and to concentrate it in the blood plasma samples. The extraction parameters such as adsorption time, pH, and the amount of sorbent in blood plasma were optimized and the capacity of loading amount (LA) for adsorbing it was determined. Moreover, a high performance liquid chromatography (HPLC)-UV detector method was validated and used for analyzing of the mentioned opioid extracted from plasma. The results showed that the limit of detection (LOD), and the limit of quantization (LOQ) for the developed MIP-SPE method were 1.24 ppb, and 3.76 ppb, respectively. Moreover, both of the MIP-, and non-imprinted polymers (NIP)-drug complexes were designed and were then optimized by the density functional theory (DFT) method. The results showed that the theoretical calculations supported the experimental data, confirming the favorability of adsorption of the drug by MIP compared to NIP.

Overview, consequences, and strategies for overcoming matrix effects in LC-MS analysis: a critical review.

The high-performance liquid chromatography-mass spectrometry (LC-MS) technique is widely applied to routine analysis in many matrices. Despite the enormous application of LC/MS, this technique is subjected to drawbacks called matrix effects (MEs) that could lead to ion suppression or ion enhancement. This phenomenon can exert a deleterious impact on the ionization efficacy of an analyte and subsequently on the important method performance parameters. LC-MS susceptibility to MEs is the main challenge of this technique in the analysis of complex matrices such as biological and food samples. Nowadays, the assessment, estimation, and overcoming of the MEs before developing a method is mandatory in any analysis. Two main approaches including the post-column infusion and post-extraction spike are proposed to determine the degree of MEs. Different strategies can be adopted to reduce or eliminate MEs depending on the complexity of the matrix. This could be done by improving extraction and clean-up methods, changing the type of ionization employed, optimization of liquid chromatography conditions, and using corrective calibration methods. This review article will provide an overview of the MEs as the Achilles heel of the LC-MS technique, the causes of ME occurrence, their consequences, and systemic approaches towards overcoming MEs during LC-MS-based multi-analyte procedures.

Design, synthesis and biological evaluation of novel indanone containing spiroisoxazoline derivatives with selective COX-2 inhibition as anticancer agents.

OBJECTIVE: A new family of 3'-(Mono, di or tri-substituted phenyl)-4'-(4-(methylsulfonyl) phenyl) spiroisoxazoline derivatives containing indanone spirobridge was designed, synthesized, and evaluated for their selective COX-2 inhibitory potency and cytotoxicity on different cell lines. METHODS: A synthetic reaction based on 1,3-dipolar cycloaddition mechanism was applied for the regiospecific formation of various spiroisoxazolines. The activity of the newly synthesized compounds was determined using in vitro cyclooxygenase inhibition assay. The toxicity of the compounds was evaluated by MTT assay. In addition, induction of apoptosis, and expression levels of Bax, Bcl-2 and caspase-3 mRNA in MCF-7 cells were evaluated following exposure to compound 9f. The docking calculations and molecular dynamics simulation were performed to study the most probable modes of interactions of compound 9f upon binding to COX-2 enzyme. RESULTS: The docking results showed that the synthesized compounds were able to form hydrogen bonds with COX-2 involving methyl sulfonyl, spiroisoxazoline, meta-methoxy and fluoro functional groups. Spiroisoxazoline derivatives containing methoxy group at the C-3' phenyl ring meta position (9f and 9g) showed superior selectivity with higher potency of inhibiting COX-2 enzyme. Furthermore, compound 9f, which possesses 3,4-dimethoxyphenyl on C-3' carbon atom of isoxazoline ring, exhibited the highest COX-2 inhibitory activity, and also displayed the most potent cytotoxicity on MCF-7 cells with an IC50 value of 0.03 ± 0.01 µM, comparable with that of doxorubicin (IC50 of 0.062 ± 0.012 µM). The results indicated that compound 9f could promote apoptosis. Also, compared to the control group, the mRNA expression of Bax and caspase-3 significantly increased, while that of Bcl-2 significantly decreased upon exposure to compound 9f which may propose the activation of mitochondrial-associated pathway as the mechanism of observed apoptosis. CONCLUSION: In vitro biological evaluations accompanied with in silico studies revealed that indanone tricyclic spiroisoxazoline derivatives are good candidates for the development of new anti-inflammatory and anticancer (colorectal and breast) agents.

Application of genetic algorithm and multivariate methods for the detection and measurement of milk-surfactant adulteration by attenuated total reflection and near-infrared spectroscopy.

BACKGROUND: The adulteration of milk by hazardous chemicals like surfactants has recently increased. It conceals the quality of the product to gain profit. As milk and milk-based products are consumed by many people, novel analytical procedures are needed to detect these adulterants. This study focused on Fourier-transform infrared (FTIR) spectroscopy equipped with an attenuated total reflection (ATR) accessory, and near-infrared (NIR) spectroscopy for the determination of milk-surfactant adulteration using a genetic algorithm (GA) coupled with multivariate methods. The model surfactant was sodium dodecyl sulfate (SDS), and its concentration varied from 1.94-19.4 gkg-1 in adulterated samples. RESULTS: Prominent peaks in the spectral range of 5500-6400 cm-1 , 1160-1260 cm-1 and 1049-1080 cm-1 may correspond to the sulfonate group in SDS. A genetic algorithm could significantly reduce the number of variables to almost one third by selecting the specific wavenumber region. Principal component analysis (PCA) for ATR and NIR data indicated separate clusters of samples in terms of the concentration level of SDS (P ≤ 0.05). Partial least squares regression (PLSR) was used to determine the maximum R2 value for ATR and NIR data for calibration, cross-validation and prediction, which were 0.980, 0.972, 0.980, and 0.970, 0.937, and 0.956 respectively. The results showed apparent differences between unadulterated and adulterated samples using partial least squares-discriminant analysis (PLS-DA), which was validated by the permutation test. CONCLUSION: The results clearly show the successful application of the proposed methods with multivariate analysis in the selection of variables, classification, clustering, and identification of the adulterant in amounts as low as 1.94 gkg-1 in milk. © 2020 Society of Chemical Industry.

Favipiravir Effects on the Control of Clinical Symptoms of Hospitalized COVID-19 Cases: An Experience with Iranian Formulated Dosage Form.

Coronavirus disease -19 (COVID-19) pandemic, caused by SARS-CoV-2, has gradually spread worldwide, becoming a major public health event. This situation requires designing a novel antiviral agent against the SARS-CoV-2; however, this is time-consuming and the use of repurposed medicines may be promising. One such medicine is favipiravir, primarily introduced as an anti-influenza agent in east world. The aim of this study was to evaluate the efficacy and safety of favipiravir in comparison with lopinavir-ritonavir in SARS-CoV-2 infection. In this randomized clinical trial, 62 patients were recruited. These patients had bilateral pulmonary infiltration with peripheral oxygen saturation lower than 93%. The median time from symptoms onset to intervention initiation was seven days. Favipiravir was not available in the Iranian pharmaceutical market, and it was decided to formulate it at the research laboratory of School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. The patients received favipiravir tablet at a dose of 1600 mg orally twice a day for day one and then 600 mg orally twice a day for days 2 to 6. In the second group, the patients received lopinavir-ritonavir combination tablet at a dose of 200/50 mg twice a day for seven days. Fever, cough, and dyspnea were improved significantly in favipiravir group in comparison with lopinavir-ritonavir group on days four and five. Mortality rate and ICU stay in both groups were similar, and there was no significant difference in this regard (P = 0.463 and P = 0.286, respectively). Chest X-ray improvement also was not significantly different between the two groups. Adverse drug reactions occurred in both groups, and impaired liver enzymes were the most frequent adverse effect. In conclusion, early administration of oral favipiravir may reduce the duration of clinical signs and symptoms in patients with COVID-19 and hospitalization period. The mortality rate also should be investigated in future clinical trials.

The Feasibility of Two Handheld Spectrometers for Meat Speciation Combined with Chemometric Methods and Its Application for Halal Certification.

Handheld visible-near-infrared (Vis-NIR) and near-infrared (NIR) spectroscopy can be cost-effective, rapid, non-destructive and transportable techniques for identifying meat species and may be valuable for enforcement authorities, retail and consumers. In this study, a handheld Vis-NIR (400-1000 nm) and a handheld NIR (900-1700 nm) spectrometer were applied to discriminate halal meat species from pork (halal certification), as well as speciation of intact and ground lamb, beef, chicken and pork (160 meat samples). Several types of class modeling multivariate approaches were applied. The presented one-class classification (OCC) approach, especially with the Vis-NIR sensor (95-100% correct classification rate), was found to be suitable for the application of halal from non-halal meat-species discrimination. In a discriminant approach, using the Vis-NIR data and support vector machine (SVM) classification, the four meat species tested could be classified with accuracies of 93.4% and 94.7% for ground and intact meat, respectively, while with partial least-squares discriminant analysis (PLS-DA), classification accuracies were 87.4% (ground) and 88.6% (intact). Using the NIR sensor, total accuracies of the SVM models were 88.2% and 81.5% for ground and intact meats, respectively, and PLS-DA classification accuracies were 88.3% (ground) and 80% (intact). We conclude that the Vis-NIR sensor was most successful in the halal certification (OCC approaches) and speciation (discriminant approaches) for both intact and ground meat using SVM.

Comparison of near-infrared (NIR) and mid-infrared (MIR) spectroscopy based on chemometrics for saffron authentication and adulteration detection.

In this work, the potential of near-infrared (NIR) and mid-infrared (MIR) spectroscopy along with chemometrics was investigated for authentication and adulteration detection of Iranian saffron samples. First, authentication of one-hundred saffron samples was examined by principal component analysis (PCA). The results showed the NIR spectroscopy can better predict the origin of samples than the MIR. Next, partial least squares-discriminant analysis (PLS-DA) was developed to detect four common plant-derived adulterants (i.e., saffron style, calendula, safflower, and rubia). In all cases, PLS-DA classification figures of merit in terms of sensitivity, specificity, error rate and accuracy were satisfactory for both NIR and MIR datasets. The built models were then successfully validated using test set and also commercial samples. Finally, partial least squares regression (PLSR) was used to estimate the amount of adulteration. In this case, only NIR showed a good performance with regression coefficients (R2) in range of 0.95-0.99.

Mitochondria-Targeted Polyamidoamine Dendrimer-Curcumin Construct for Hepatocellular Cancer Treatment.

Mitochondrial malfunction plays a crucial role in cancer development and progression. Cancer cells show a substantially higher mitochondrial activity and greater mitochondrial transmembrane potential than normal cells. This concept can be exploited for targeting cytotoxic drugs to the mitochondria of cancer cells using mitochondrial-targeting compounds. In this study, a polyamidoamine dendrimer-based mitochondrial delivery system was prepared for curcumin using triphenylphosphonium ligands to improve the anticancer efficacy of the drug in vitro and in vivo. For the in vitro evaluations, various methods, such as viability assay, confocal microscopy, flow cytometry, reactive oxygen species (ROS), and real-time polymerase chain reaction analyses, were applied. Our findings showed that the targeted-dendrimeric curcumin (TDC) could successfully deliver and colocalize the drug to the mitochondria of the cancer cells, and selectively induce a potent apoptosis and cell cycle arrest at G2/M. Moreover, at a low curcumin dose of less than 25 µM, TDC significantly reduced adenosine triphosphate and glutathione, and increased the ROS level of the isolated rat hepatocyte mitochondria. The in vivo studies on the Hepa1-6 tumor-bearing mice also indicated a significant tumor suppression effect and the highest median survival days (Kaplan-Meier survival estimation and log-rank test) after treatment with the TDC construct compared to the free curcumin and untargeted construct. Besides its targeted nature and safety, the expected improved solubility and stability represent the prepared targeted-dendrimeric construct as an up-and-coming candidate for cancer treatment. The results of this study emphasize the promising route of mitochondrial targeting as a practical approach for cancer therapy, which can be achieved by optimizing the delivery method.

Two novel anticancer compounds with minimum cardiotoxic property.

BACKGROUND: Although two novel synthesized compounds with tri-aryl structures; 3-(4-chlorophenyl)-5-(4-fluorophenyl)-4-phenyl-4,5-dihydro-1,2,4-oxadiazole (A) and 3,5-bis-(4-chlorophenyl)-4-phenyl-4,5-dihydro-1,2,4-oxadiazole (B) have been previously demonstrated to possess remarkable anti-breast cancer activity, their cardiotoxicity remains a major concern due to their mechanism of action. To address this concern, we assessed the ability of these compounds to cause toxicity towards H9c2 cardiomyocytes as an in vitro model of cardiotoxicity. METHODS: Cytotoxic activity of both compounds was explored in vitro on H9c2 cells using MTT assay. Annexin V/PI method, intracellular ROS determination and mitochondrial membrane potential assay were applied to elucidate the mechanism of action of the cell death. RESULTS: MTT assay revealed a concentration- and time-dependent cardiotoxicity. Findings of apoptosis by double staining with annexin V and propidium iodide divulged no cell death including apoptosis and necrosis at the concentration that were effective to inhibit cancer cells proliferation (10 µM) at 24 and 48 h. Furthermore, flow cytometric measurement of membrane potential and ROS determination using DCFH-DA verified the safe concentration of the compounds against H9c2 cells with no cardiotoxic effect. However, the higher concentration of the compounds could induce cell death through ROS-mediated mitochondrial dysfunction. CONCLUSIONS: Altogether, the results represented two novel chemical molecules possessing anti-breast cancer activity with minimum cardiac side effect.

Global Sequence Analysis and Expression of Azurin Gene in Different Clinical Specimens of Burn Patients with Pseudomonas aeruginosa Infection.

AIM: The purpose of this study was to analyze the sequence of azurin gene in relation to its expression in Pseudomanas aeruginosa strains isolated from different clinical specimens of burn patients. Moreover, in silico sequence analysis of azurin gene using globally reported sequences was intended. MATERIALS AND METHODS: Fifty-nine multidrug-resistant P. aeruginosa isolates were selected from different clinical specimens of patients suffering from burn wound infections in two university hospitals and subjected to antibacterial susceptibility testing. The frequency and genetic diversity of the azurin gene was determined by polymerase chain reaction (PCR) and Sanger sequencing. The azurin gene sequences were compared with the sequence data from other countries. The expression level of azurin gene in P. aeruginosa isolates with different azurin sequences from different clinical specimens was evaluated by real-time PCR. RESULTS AND CONCLUSION: About 98%-100% of the isolates were resistant to gentamicin, tobramycin, cefoxitin, ciprofloxacin, amikacin, and imipenem, while 100% and 23.9% of the isolates were susceptible to colistin and ceftazidime, respectively. Only eight point mutations were detected with amino acid substitutions in only two positions (81 and 102). In global analysis, 93% of strains showed missense mutation at positions 81 (alanine to threonine). The majority (81%) of Iranian strains were allocated to two major clusters distinct from the rest of world, which may suggest that strains from Iran have made a distinct genetic stockpile through point mutations which has established them separate from the other counties. However, 19% were distributed in different clusters together with the strains from different countries of North and South America, Europe, South and East Asia. The expression level of the azurin gene was statistically higher in the isolates collected from the blood of burns patients with systemic infection compared to the isolates collected from other specimens (wound, catheter and tissue), which shows a positive correlation between azurin gene expression and increased pathogenicity and capability for dissemination. This study may open new insight about azurin genetic variation and significance in P. aeruginosa pathogenesis.

Combining multivariate image analysis with high-performance thin-layer chromatography for development of a reliable tool for saffron authentication and adulteration detection.

In this work, high-performance thin-layer chromatography (HPTLC) coupled with multivariate image analysis (MIA) is proposed as a fast and reliable tool for authentication and adulteration detection of Iranian saffron samples based on their HPTLC fingerprints. At first, the secondary metabolites of saffron were extracted using ultrasonic-assisted solvent extraction (UASE) which was optimized using central composite design (CCD). Next, the RGB coordinates of HPTLC images were used for estimation of saffron origin based on principal component analysis (PCA). The PCA scores plot showed that saffron samples were clustered into two clear-cut groups which was 92% matched with the geographical origins of the samples. In the next step, common plant-derived adulterants of saffron including safflower, saffron style, calendula, and rubia were investigated with MIA analysis of HPTLC images using partial least squares-discriminant analysis (PLS-DA) at 5-35% (w/w) levels. The PLS-DA results showed proper classification of saffron and adulterants with sensitivity 99.14%, specificity 96.94%, error rate 1.96% and accuracy 98.04. Also, the effect of HPTLC injection volume on the performance of the proposed strategy was evaluated. The ability of the proposed method was then investigated by analyzing two additional sample sets including mixed samples of four plant-derived adulterants and adulterated commercial samples. Sensitivity and specificity of this model were 100% which confirmed its validity.