Exploring the biological application of Penicillium fimorum-derived silver nanoparticles: In vitro physicochemical, antifungal, biofilm inhibitory, antioxidant, anticoagulant, and thrombolytic performance.

This study showed the anti-candida, biofilm inhibitory, antioxidant, anticoagulant, and thrombolytic properties of biogenic silver nanoparticles (AgNPs) fabricated by using the supernatant of Penicillium fimorum (GenBank accession number OQ568180) isolated from soil. The biogenic AgNPs were characterized by using different analytical techniques. A sharp surface plasmon resonance (SPR) peak of the colloidal AgNPs at 429.5 nm in the UV-vis spectrum confirmed the fabrication of nanosized silver particles. The broth microdilution assay confirmed the anti-candida properties of AgNPs with a minimum inhibitory concentration (MIC) of 4 µg mL-1. In the next step, the protein and DNA leakage assays as well as reactive oxygen species (ROS) assay were performed to evaluate the possible anti-candida mechanisms of AgNPs representing an increase in the total protein and DNA of supernatant along with a climb-up in ROS levels in AgNPs-treated samples. Flow cytometry also confirmed a dose-dependent cell death in the AgNPs-treated samples. Further studies also confirmed the biofilm inhibitory performance of AgNPs against Candia albicans. The AgNPs at the concentrations of MIC and 4*MIC inhibited 79.68 ± 14.38% and 83.57 ± 3.41% of biofilm formation in C. albicans, respectively. Moreover, this study showed that the intrinsic pathway may play a significant role in the anticoagulant properties of AgNPs. In addition, the AgNPs at the concentration of 500 µg mL-1, represented 49.27%, and 73.96 ± 2.59% thrombolytic and DPPH radical scavenging potential, respectively. Promising biological performance of AgNPs suggests these nanomaterials as a good candidate for biomedical and pharmaceutical applications.

Challenges for the determination of spiramycin in aqueous matrices using LC-MS/MS: evidence for the solvent intrusion on the molecule integrity.

Liquid chromatography-tandem mass spectroscopy (LC-MS/MS) is an accurate and specific technique for drug residue analysis in different matrices. The high specificity and sensitivity of the multiple reaction monitoring (MRM) approach for detecting drugs such as aldehydes, which have the potential to change mass during the sample preparation phase, becomes a drawback during the analysis process. In this study, concerns about the intrusion of solvent molecules into spiramycin's chemical structure as an aldehydic drug as well as the stability of spiramycin in the milk matrix were addressed. Furthermore, the binding sites where the solvent molecules could bind to spiramycin molecules were investigated through nuclear magnetic resonance (NMR) spectroscopy. It was revealed that water, ethanol, and methanol as protic solvents can add to the formyl group of spiramycin molecules during standard solutions preparation while there was no evidence for the addition of acetonitrile and dimethyl sulfoxide (aprotic solvents). In addition, as time passed, the peak area of spiramycin decreased either in the spiked aqueous sample or milk sample while an increase in the peak area of H2O-bound spiramycin was observed. After 96 h, more than 90% of spiramycin was converted to H2O-bound spiramycin. In conclusion, we can propose the use of aprotic solvents for the preparation of spiramycin standard solutions especially when the prepared solutions are not used instantly. Moreover, ion transitions for both spiramycin and its H2O-added form (843.6 m/z to 173.9 m/z and 861.5 m/z to 173.9 m/z, respectively) should be considered for the accurate quantification of spiramycin residue in aqueous samples such as milk.

Comparison of Portable and Benchtop Near-Infrared Spectrometers for the Detection of Citric Acid-adulterated Lime Juice: A Chemometrics Approach.

Background: Since the incidence of food adulteration is rising, finding a rapid, accurate, precise, low-cost, user-friendly, high-throughput, ruggedized, and ideally portable method is valuable to combat food fraud. Near-infrared spectroscopy (NIRS), in combination with a chemometrics-based approach, allows potentially rapid, frequent, and in situ measurements in supply chains. Methods: This study focused on the feasibility of a benchtop Fourier-transformation-NIRS apparatus (FT-NIRS, 1000 - 2500 nm) and a portable short wave NIRS device (SW-NIRS, 740 - 1070 nm) for the discrimination of genuine and citric acid-adulterated lime juice samples in a cost-effective manner following chemometrics study. Results: Principal component analysis (PCA) of the spectral data resulted in a noticeable distinction between genuine and adulterated samples. Wavelengths between 1100 - 1400 nm and 1550 - 1900 nm were found to be more important for the discrimination of samples for the benchtop FT-NIRS data, while variables between 950 - 1050 nm contributed significantly to the discrimination of samples based on the portable SW-NIRS data. Following partial least squares discriminant analysis (PLS-DA) as a discriminant model, standard normal variate (SNV) or multiplicative scatter correction (MSC) transformation of benchtop FT-NIRS data and SNV in combination with the second derivative transformation of portable SW-NIRS data on the training set delivered equal accuracy (94%) in the prediction of the test set. In the soft independent modeling of class analogy (SIMCA) as a class-modeling approach, the overall performances of generated models on the auto-scaled data were 98% and 94.5% for benchtop FT-NIRS and portable SW-NIRS, respectively. Conclusions: As a proof of concept, NIRS technology coupled with appropriate multivariate classification models enables fast detection of citric acid-adulterated lime juices. In addition, the promising results of portable SW-NIRS combined with SIMCA indicated its use as a screening tool for on-site analysis of lime juices at various stages of the food supply chain.

Design, synthesis, and preliminary pharmacological evaluation of novel thiazolidinone derivatives as potential benzodiazepine agonists.

Thiazolidinones are well-known heterocycles that demonstrate promising biological effects such as anticonvulsant activity. Hybridization of these chemicals with scaffold, which has necessary pharmacophores for binding to the benzodiazepine receptors, can prompt a novel structure possessing extensive anticonvulsant effects. In this study, novel derivatives of thiazolidinone as new benzodiazepine agonists were designed, synthesized, and biologically evaluated. Compound 5h, 4-chloro-2-(2-fluorophenoxy)-N-(4-oxo-2-(p-tolyl)thiazolidin-3-yl)benzamide, exhibited considerable anticonvulsant activity, proper sedative-hypnotic effect, no memory impairment, and no muscle relaxant effect. The pharmacological effects of the designed compounds were antagonized by flumazenil, which confirmed the benzodiazepine receptors' involvement in their biological effects. Based on in silico calculations of ADME properties of our novel compounds, they could be active oral agents potentially. In this study, we designed novel structures by the hybridization of thiazolidinone moiety with scaffold which has necessary pharmacophores for binding to the benzodiazepine receptors. The results are very promising for developing new lead compounds as benzodiazepine agonists possess anticonvulsant effects.

Sedative-hypnotic, anxiolytic and possible side effects of Salvia limbata C. A. Mey. Extracts and the effects of phenological stage and altitude on the rosmarinic acid content.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia limbata C. A. Mey. (Persian name: Maryam Goli-e-labeh dar) has been used for treating central nervous disorders such as insomnia, anxiety and depression in Persian traditional medicine. S. limbata is known for its pharmacological activities which could be at least in a part, upon the presence of rosmarinic acid (RA). However, the sedative-hypnotic effect, anxiolytic activity, possible side effects, and the mechanism of action of S. limbata extract has not yet been examined. AIM OF THE STUDY: In the current study the sedative-hypnotic effect, anxiolytic activity, possible side effects, and the mechanism of action of S. limbata extracts were evaluated. Besides, the effects of altitude and phenological stage on the RA content of S. limbata were investigated. MATERIALS AND METHODS: Sedative-hypnotic and anxiolytic effects were evaluated through the pentobarbital induced loss of righting reflex test and open field test, respectively. Flumazenil was used to reveal the mechanism of action. Possible side effects were investigated in the passive avoidance and grip strength tests. Besides, the effects of altitude and phenological stage (vegetative, flowering, and seed setting) on the RA content of S. limbata were evaluated using reversed-phase high-performance liquid chromatography (RP-HPLC). RESULTS: Following behavioral tests, sedative-hypnotic and anxiolytic effects were observed. Since the observed effects were reversed by flumazenil and no side effect on the memory and muscle strength was reported, modulation of the α1-containing GABA-A receptors could be proposed as one of the involved mechanisms. According to the RP-HPLC analysis, harvesting S. limbata in the vegetative stage at the altitude of 2500 m led to the highest content of RA (8.67 ± 0.13 mg/g dry matter). Among different extract of the plant samples collected in the vegetative stage at the altitude of 2500 m, the hydroalcoholic extract showed the highest rosmarinic acid content. CONCLUSION: The obtained results help to find the optimum situation to gain the highest content of RA as well as the pharmacological activity that could be economically important for the pharmaceutical industries.

A comparison of artificial intelligence techniques for predicting hyperforin content in Hypericum perforatum L. in different ecological habitats.

Hyperforin, a major bioactive constituent of Hypericum concentration, is impacted by various phenological phases and soil characteristics. We aimed to design a model predicting hyperforin content in Hypericum perforatum based on different ecological and phenological conditions. We employed artificial intelligence modeling techniques including multilayer perceptron (MLP), radial basis function (RBF), and support vector machine (SVM) to examine the factors critical in predicting hyperforin content. We found that the MLP model (R 2 = .9) is the most suitable and precise model compared with RBF (R 2 = .81) and SVM (R 2 = .74) in predicting hyperforin in H. perforatum based on ecological conditions, plant growth, and soil features. Moreover, phenological stages, organic carbon, altitude, and total N are detected in sensitivity analysis as the main factors that have a considerable impact on hyperforin content. We also report that the developed graphical user interface would be adaptable for key stakeholders including producers, manufacturers, analytical laboratory managers, and pharmacognosists.

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.

Changes in the essential oil content and composition of Salvia limbata C.A. Mey at different growth stages and altitudes.

Salvia limbata is of great importance to the pharmaceutical industry owing to its various biological effects. Therefore, it is important to investigate the main factors that affect its essential oil composition. Although some investigations have been performed with regard to the phytochemistry of S. limbata, this study investigates, for the first time, the effect of growth stage and altitude on the content and chemical composition of essential oil extracted from S. limbata. For this purpose, the essential oil was extracted from 45 air-dried samples by hydrodistillation and analyzed by GC-MS and GC-flame methods. The highest content of essential oil was obtained from aerial parts in the vegetative stage at an altitude of 1500 m (0.86% v/w). Our findings show that the vegetative stage at 1500 m is the optimal harvest time to extract the highest content of oil while the highest content of monoterpenes (including α-pinene and ß-pinene) could be obtained in the same phenological stage at 2000 m. By contrast, the content of sesquiterpenes increased to the highest values in the ripening stage at 1500 and 2500 m. The results of this study help to find the optimal conditions to obtain the highest content of S. limbata essential oil, but additional studies are warranted.

Prediction of hypericin content in Hypericum perforatum L. in different ecological habitat using artificial neural networks.

BACKGROUND: Hypericum is an important genus in the family Hypericaceae, which includes 484 species. This genus has been grown in temperate regions and used for treating wounds, eczema and burns. The aim of this study was to predict the content of hypericin in Hypericum perforatum in varied ecological and phenological conditions of habitat using artificial neural network techniques [MLP (Multi-Layer Perceptron), RBF (Radial Basis Function) and SVM (Support Vector Machine)]. RESULTS: According to the results, the MLP model (R2 = 0.87) had an advantage over RBF (R2 = 0.8) and SVM (R2 = 0.54) models and it was relatively accurate in predicting hypericin content in H. perforatum based on the ecological conditions of site including soil types, its characteristics and plant phenological stages of habitat. The results of sensitivity analysis revealed that phenological stages, hill aspects, total nitrogen, altitude and organic carbon are the most influential factors that have an integral effect on the content of hypericin. CONCLUSIONS: The designed graphical user interface will help pharmacognosist, manufacturers and producers of medicinal plants and so on to run the MLP model on new data to easily discover the content of hypericin in H. perforatum by entering ecological conditions of site, soil characteristics and plant phenological stages.

Novel Application of Near-infrared Spectroscopy and Chemometrics Approach for Detection of Lime Juice Adulteration.

The aim of this study is to investigate the novel application of a handheld near infra-red spectrophotometer coupled with classification methodologies as a screening approach in detection of adulterated lime juices. For this purpose, a miniaturized near infra-red spectrophotometer (Tellspec®) in the spectral range of 900-1700 nm was used. Three diffuse reflectance spectra of 31 pure lime juices were collected from Jahrom, Iran and 25 adulterated juices were acquired. Principal component analysis was almost able to generate two clusters. Partial least square discriminant analysis and k-nearest neighbors algorithms with different spectral preprocessing techniques were applied as predictive models. In the partial least squares discriminant analysis, the most accurate prediction was obtained with SNV transforming. The generated model was able to classify juices with an accuracy of 88% and the Matthew's correlation coefficient value of 0.75 in the external validation set. In the k-NN model, the highest accuracy and Matthew's correlation coefficient in the test set (88% and 0.76, respectively) was obtained with multiplicative signal correction followed by 2nd-order derivative and 5th nearest neighbor. The results of this preliminary study provided promising evidence of the potential of the handheld near infra-red spectrometer and machine learning methods for rapid detection of lime juice adulteration. Since a limited number of the samples were used in the current study, more lime juice samples from a wider range of variability need to be analyzed in order to increase the robustness of the generated models and to confirm the promising results achieved in this study.

Design, synthesis, and pharmacological evaluation of novel 1,2,4-triazol-3-amine derivatives as potential agonists of GABAA subtype receptors with anticonvulsant and hypnotic effects.

In the current study, a series of novel 1,2,4-triazol-3-amine derivatives were designed, synthesized, and biologically evaluated in vivo for their anticonvulsant and hypnotic effects in the pentylenetetrazole (PTZ)-induced seizures, maximal electroshock (MES)-induced seizures, and pentobarbital-induced sleeping tests. Furthermore, the possible side effects of the most potent compounds on the memory, motor coordination, and muscle strength were evaluated in passive avoidance, rotarod, and grip strength tests, respectively. The designed compounds with the main benzodiazepine pharmacophores including aromatic ring and proton accepting group completely mimiced the structure of zolpidem as an α1-selective agonist of GABAA receptor. Compounds 5c (ED50 ≈ 52.5 mg/kg) and 5 g (ED50 ≈ 16.5 mg/kg) in the PTZ test were the most potent compounds among the designed compounds. In the MES test, the observed ED50s for compounds 5c and 5 g were reduced to around 11.8 mg/kg and 10.5 mg/kg, respectively. The considerable hypnotic effect in a dose-dependent manner was observed following the administration of newly synthesized compounds. In all experiments administration of flumazenil as an antagonist of benzodiazepines receptor fully antagonized observed effects which indicated the involvement of GABAA receptors. Since there was no negative effect on memory, motor coordination, and muscle strength following the administration of compounds 5c and 5g as the most potent compounds, it could be concluded that the novel compounds most likely act through α1-containing GABAA receptors and possess no affinity for α5-containing receptors. The newly designed compounds could be considered as leading compounds in synthesizing novel GABAA receptor agonists with minimum side effects.

MLR and ANN Approaches for Prediction of Synthetic/Natural Nanofibers Diameter in the Environmental and Medical Applications.

Fiber diameter plays an important role in the properties of electrospinning of nanofibers. However, one major problem is the lack of a comprehensive method that can link processing parameters to nanofibers' diameter. The objective of this study is to develope an artificial neural network (ANN) modeling and multiple regression (MLR) analysis approaches to predict the diameter of nanofibers. Processing parameters, including weight ratio, voltage, injection rate, and distance, were considered as independent variables and the nanofiber diameter as the dependent variable of the ANN model. The results of ANN modeling, especially its high accuracy (R2 = 0.959) in comparison with MLR results (R2 = 0.564), introduced the prediction the diameter of nanofibers model (PDNFM) as a comparative model for predicting the diameter of poly (3-caprolactone) (PCL)/gelatin (Gt) nanofibers. According to the result of sensitivity analysis of the model, the values of weight ratio, distance, injection rate, and voltage, respectively, were identified as the most significant parameters which influence PDNFM.

Neuro-Behavioral Profile and Toxicity of the Essential Oil of Dorema ammoniacum Gum as an Anti-seizure, Anti-nociceptive, and Hypnotic Agent with Memory-enhancing Properties in D-Galactose Induced Aging Mice.

In this study, we focused on the neuro-behavioral profile, toxicity, and possible mechanisms of action of Dorema ammoniacum gum essential oil (DAG-EO). For this purpose, passive avoidance and Y-maze tests were performed to evaluate the potential effect of DAG-EO in the attenuation of memory impairment induced by 49 days administration of D-galactose and acute injection of scopolamine. Anticonvulsant and anti-nociceptive activities of DAG-EO were evaluated in the pentylenetetrazole and maximal electroshock-induced models of seizure and acetic acid-induced writhing tests, respectively. To find the possible mechanism of action, flumazenil and naloxone were used. Furthermore, the possible side effects were determined in the open field, grip strength, and rotarod tests. Our findings supported that 7-day administration of DAG-EO (50 and 100 mg/kg) improves memory impairment induced following administration of D-galactose and scopolamine. It was also revealed that DAG-EO possesses a dose-dependent sedative-hypnotic (100 mg/kg), anticonvulsant (ED50 ≈ 170 mg/kg), and anti-nociceptive (ED50 ≈ 175 mg/kg) activities possibly mediated via directly and/or indirectly modulation of GABAA and opioid receptors. No side effect was observed except muscle relaxation which was less than that of diazepam. The output of this study confirms anti-seizure, anti-nociceptive, sedative-hypnotic, and memory-enhancing properties of DAG-EO by modulation of GABAA receptors.

Evaluation and comparison of the antidepressant-like activity of Artemisia dracunculus and Stachys lavandulifolia ethanolic extracts: an in vivo study.

Several studies have supported the preventive and therapeutic values of phenolic compounds including chlorogenic acid, syringic acid, vanillic acid, ferulic acid, caffeic acid, luteolin, rutin, catechin, kaempferol, and quercetin in mental disorders. Since these secondary metabolites are reported as the phenolic compounds of Artemisia dracunculus (A. dracunculus) and Stachys lavandulifolia (S. lavandulifolia), the main aim of this study was the evaluation and comparison of the phenolic contents, flavonoids, and antidepressant-like activity of Artemisia dracunculus with Stachys lavandulifolia. Antidepressant-like activity of the extracts was evaluated in the forced swimming test (FST) and the tail suspension test (TST). Moreover, the open field test was conducted to evaluate the general locomotor activity of mice following treatment with the extracts. Since phenolic compounds and flavonoids play main roles in pharmacological effects, the phenolic and flavonoid contents of the extracts were measured. Though significant difference between the phenolic contents of the extracts was not observed, but S. lavandulifolia exhibited higher flavonoid contents. Animal treatment with extracts decreased the immobility times in both FST and TST compared to the vehicle group without any significant effect on the locomotor activity of animals. Also, S. lavandulifolia at 400 mg/kg showed higher potency in both tests compared to A. dracunculus. Our results provided promising evidence on the antidepressant-like activity of both extracts which could be related to flavonoids as the main components of the extracts, but more studies need to be conducted to specify the main compounds and the mechanisms involved in the observed effects.

An In-Vivo Study on Anticonvulsant, Anxiolytic, and Sedative-Hypnotic Effects of the Polyphenol-Rich Thymus Kotschyanus Extract; Evidence for the Involvement of GABAA Receptors.

Antidepressant-like activity of T. kotschyanus has been recently reported by scientists but insufficient attention has been so far devoted to T. kotschyanus, and there is a lack of information on the other neurobehavioral effects and side effects of this species. In the current study, the anticonvulsant, anxiolytic, and sedative-hypnotic, effects of Thymus kotschyanus extract on male NMRI mice were evaluated using pentylenetetrazole, maximal electroshock, elevated plus maze, and pentobarbital-induced sleeping tests. Since phenolic compounds and flavonoids have main roles in pharmacological effects of most plant extracts, the phenolic and flavonoid contents of the extract were measured with Folin-Ciocalteu and AlCl3 reagents. Acute toxicity, passive avoidance, and open field tests were carried out to assess the toxicity of the extract. To find out the possible mechanism of action, flumazenil as the specific GABAA receptor antagonist was used. Anticonvulsant and hypnotic effects of the extract were observed at 400 and 600 mg/kg. The extract at the dose of 200 mg/kg revealed significant anxiolytic effects, but it did not show any adverse effects on learning and memory at all the tested doses. Results of this study indicate that Thymus kotschyanus extract has anticonvulsant , anxiolytic, and hypnotic effects, which are likely related to the ability of some phenolic compounds to activate α1-containing GABAA receptors but more experiments still need to be carried out in order to find the exact mechanism, active component, and the toxicity of the Thymus kotschyanus extract.

Novel 4-thiazolidinone derivatives as agonists of benzodiazepine receptors: Design, synthesis and pharmacological evaluation.

A new series of 4-chloro-N-(2-(substitutedphenyl)-4-oxothiazolidin-3-yl)-2-phenoxybenzamide derivatives were designed, synthesized and biologically evaluated as anticonvulsant agents. The designed compounds have the main essential functional groups for binding to the benzodiazepine receptors and 4-thiazolidinone ring as an anticonvulsant pharmacophore. Some of the new synthesized compounds showed considerable anticonvulsant activity in electroshock and pentylenetetrazole-induced lethal convulsion tests. Compound 5i, 4-chloro-N-(2-(4-methoxyphenyl)-4-oxothiazolidin-3-yl)-2-phenoxybenzamide, with the best activity was selected for evaluation of other benzodiazepine pharmacological effects. This compound induced significant sedative-hypnotic activity. However, it does not impair the learning and memory in the experimental condition. Flumazenil was able to antagonize the sedative-hypnotic and anticonvulsant effects of compound 5i indicating that benzodiazepine receptors are highly involved in the pharmacological properties of the novel compounds.

Design, synthesis, pharmacological evaluation, and docking study of new acridone-based 1,2,4-oxadiazoles as potential anticonvulsant agents.

A number of acridone-based oxadiazoles 11a-n have been synthesized and evaluated for their anticonvulsant activity against pentylenetetrazole (PTZ)- and maximal electroshock (MES)-induced seizures in mice. Also, their neurotoxicity was evaluated by the rotarod test. Most of the compounds exhibited better anticonvulsant activity and higher safety respect to the standard drug, phenobarbital. Among the tested derivatives, compounds 11l with ED50 value of 2.08 mg/kg was the most potent compound in the PTZ test. The anticonvulsant effect of compound 11l was blocked by flumazenil, suggesting the involvement of benzodiazepine (BZD) receptors in the anticonvulsant activity of prototype compound 11l. Also, docking study of compound 11l in the BZD-binding site of GABAA receptor confirms possible binding of compound 11l with BZD receptors.