The anti-biofilm activity of hydrogen peroxide against Escherichia coli strain FL-Tbz isolated from a pharmaceutical water system.

Biofilms are considered a significant reason for the failure of disinfection strategies in industrial water systems due to their resistance to antimicrobial agents. This study is designed to investigate the anti-biofilm activity of hydrogen peroxide (H2O2) at combinations of temperatures and contact times. For this purpose, an in vitro microtiter plate (MTP)-based model system was used for biofilm formation using Escherichia coli (E. coli) strain FL-Tbz isolated from the water system of a pharmaceutical plant. To investigate the anti-biofilm activity of H2O2, it was added at different concentrations (2-7% v/v) to biofilms and incubated at different temperatures (20-60 °C) for 10-40 min to find effective conditions to eradicate biofilms. Maximum biofilms were formed when bacterial suspensions were incubated at 37 °C for 96 h. The rate of biofilm formation using an environmental isolate was higher than that of standard strain. H2O2 at concentrations of ≥6.25% (v/v) at temperatures of ≥40 °C incubated for ≥25 min significantly eradicated the biofilms.

Dispersive micro-solid-phase extraction of aflatoxins from commercial soy milk samples using a green vitamin-based metal-organic framework as an efficient sorbent followed by high performance liquid chromatography-tandem mass spectrometry determination.

In the current work, for the first time, a vitamin-based metal-organic framework constructed from cobalt ions and vitamin B3 has been used as a sorbent in dispersive micro-solid-phase extraction. The proposed method was used to extract and enrich aflatoxins (B1, B2, G1, and G2) from soy milk samples before their quantification by high performance liquid chromatography-tandem mass spectrometry. In this work, first the metal-organic framework was synthesized and characterized using techniques such as X-ray diffraction, Fourier transform infrared spectrophotometry, scanning electron microscopy, nitrogen adsorption/desorption, and energy-dispersive X-ray spectroscopy. Then it was used as an efficient sorbent in the proposed dispersive micro-solid-phase extraction. For this purpose, after precipitating the proteins of soy milk sample with the aid of trichloroacetic acid, the supernatant phase was taken, mixed with the synthesized sorbent, and vortexed. After centrifuging, the analytes loaded on the adsorbent were eluted with methanol to transfer them into an organic phase which was compatible with the subsequently employed separation system. The adsorption capacity of the synthesized MOF for aflatoxins B1, B2, G1, and G2 were 0.77, 0.83, 0.70, and 0.54 mg g-1, respectively. Under the best experimental situations, satisfactory outcomes including acceptable extraction recoveries (64-75%), low limits of detection (11.3-48.2 ng L-1) and quantification (42.8-161.6 ng L-1), and good repeatability (relative standard deviations equal or less than 4.0 and 4.7% for intra- and inter- day precisions, respectively) were obtained. In addition, green synthesis of the metal-organic framework (using vitamin B3 as a linker, water as the reaction solvent, and mild conditions) and usage low amount or volume of the adsorbent and organic solvents during the extraction process were the other beneficial aspects of this work which caused the suggested analytical method to be environmentally friendly.

Targeting Multidrug Resistance With Antimicrobial Peptide-Decorated Nanoparticles and Polymers.

As a category of small peptides frequently found in nature, antimicrobial peptides (AMPs) constitute a major part of the innate immune system of various organisms. Antimicrobial peptides feature various inhibitory effects against fungi, bacteria, viruses, and parasites. Due to the increasing concerns of antibiotic resistance among microorganisms, development of antimicrobial peptides is an emerging tool as a favorable applicability prospect in food, medicine, aquaculture, animal husbandry, and agriculture. This review presents the latest research progress made in the field of antimicrobial peptides, such as their mechanism of action, classification, application status, design techniques, and a review on decoration of nanoparticles and polymers with AMPs that are used in treating multidrug resistance. Lastly, we will highlight recent progress in antiviral peptides to treat emerging viral diseases (e.g., anti-coronavirus peptides) and discuss the outlook of AMP applications.

Safety issues of nanomaterials for dermal pharmaceutical products.

Nanomaterials (NMs) have favorable application in the medicine area, specifically in regard to the carry of pharmaceutical ingredients to provide targeted drug delivery systems. The skin is an excellent route for the delivery of pharmaceutical nano-transporters for skin-related applications. The physicochemical properties of nanomaterials such as size, hydrophobicity, loading capacity, charge and weight are vital for a skin penetrating system. Many nanocarriers such as polymeric nanoparticles, inorganic nanomaterials and, lipid nanostructures have been utilized for dermal delivery of active ingredients and others such as carbon nanotubes and fullerenes require more examination for future application in the skin-related area. Some negative side effects and nano-cytotoxicity of nanomaterials require special attention while investigating different nanomaterials for medicinal applications. Then, in the current review, we had a view on the safety issues of nanomaterials for dermal pharmaceutical products.

Application of calcium oxide as an efficient phase separation agent in temperature-induced counter-current homogenous liquid-liquid extraction of aflatoxins from dried fruit chips followed by high-performance liquid chromatography-tandem mass spectrometry determination.

A temperature-induced counter-current homogenous liquid-liquid extraction procedure performed in a burette has been proposed for the isolation of aflatoxins B1, B2, G1, and G2 from various fruit chip samples. In this method, a homogenous solution of deionized water and cyclohexylamine is added to the solid sample and the resulted mixture is vortexed. In the following, the liquid phase is taken and passed through the burette filled with a mixture of calcium oxide (as a phase separation agent) and sand (to avoid clumping the calcium oxide). By doing so, the temperature of the solution is increased by hydration of calcium oxide and consequently, the homogenous state is broken and the aflatoxins are migrated into the resulted tiny droplets of cyclohexylamine. This phase is collected on the top of the solution owing to its low density with respect to an aqueous solution. Numerous parameters which can affect the efficiency of the suggested approach were evaluated and under the best situations, great repeatability, low limits of determination and quantification, and high extraction recoveries were acquired. In the end, the suggested approach was employed for the quantification of the selected aflatoxins in various fruit chips samples marketed in Tabriz City, Iran.

A novel method for the development of plasmid DNA-loaded nanoliposomes for cancer gene therapy.

We aimed to develop a simple yet novel method to prepare plasmid DNA-loaded nanoliposomes for cancer gene therapy. Murine interleukin-12 (mIL-12) pDNA-loaded nanoliposomes were prepared via novel freeze-drying of a monophase solution method. The physicochemical characteristics, cytotoxicity, and transfection efficiency of the prepared nanoliposomes in murine CT-26 colon carcinoma cells were evaluated. Furthermore, tumor progression and survival rate in CT-26 colon carcinoma-bearing BALB/c mice subsequent to direct intratumoral injections were investigated over a period of 40 days. Using this preparation method, nanoliposomes with particle size of around 300 nm and zeta potential of 96.5 mV were obtained. The transmission electron microscope results showed that the liposomes were nano-sized and almost spherical. The agarose gel retardation assay revealed the pDNA encapsulation in the nanoliposomes. The nanoliposomes with 72.4% encapsulation efficiency and low cell toxicity could significantly improve mIL-12 expression by approximately 25-fold relative to the naked mIL-12 pDNA. There was a significant tumor growth inhibition after repeated injections of mIL-12 pDNA-loaded nanoliposomes. This is the first study on the freeze-drying of a monophase solution method as a simple yet novel technique for the preparation of pDNA-loaded nanoliposomes. Given the ease of preparation method and promising in vitro and in vivo characteristics, this investigation demonstrates advances in pDNA lipid formulation for cancer gene therapy.

Safety and Toxicity Issues of Therapeutically Used Nanoparticles from the Oral Route.

The emerging science of nanotechnology sparked a research attention in its potential benefits in comparison to the conventional materials used. Oral products prepared via nanoparticles (NPs) have garnered great interest worldwide. They are used commonly to incorporate nutrients and provide antimicrobial activity. Formulation into NPs can offer opportunities for targeted drug delivery, improve drug stability in the harsh environment of the gastrointestinal (GI) tract, increase drug solubility and bioavailability, and provide sustained release in the GI tract. However, some issues like the management of toxicity and safe handling of NPs are still debated and should be well concerned before their application in oral preparations. This article will help the reader to understand safety issues of NPs in oral drug delivery and provides some recommendations to the use of NPs in the drug industry.

Air-assisted liquid-liquid microextraction of total 3-monochloropropane-1,2-diol from refined edible oils based on a natural deep eutectic solvent and its determination by gas chromatography-mass spectrometry.

In this paper, a fast, sensitive, and selective sample preparation procedure was presented for the determination of 3-monochloropropane-1,2-diol (3-MCPD) in refined edible oils using gas chromatography-mass spectrometry. In this method, firstly, the sample lipids and analyte fatty esters are saponified by sodium hydroxide under sonication. After that the analyte was derivatized using phenylboronic acid (as the derivatization agent) and the obtained derivative was extracted during an air-assisted liquid-liquid microextraction procedure (AALLME). Six different deep eutectic solvents (DESs) were prepared as the extraction solvents and the most effective extraction for 3-MCPD was obtained in the presence of a natural DES (NDES) consisting of choline chloride (ChCl)-acetic acid (AcOH). Important variables such as sodium hydroxide concentration and volume, sonication time, temperature, extraction solvent type and volume, and phenylboronic acid concentration and volume have been optimized. Using the optimum conditions, broad linear range (0.88-1000 ng g-1), suitable coefficient of determination (0.995), and low limits of detection (0.26 ng g-1) and quantification (0.88 ng g-1) were obtained. Relative standard deviations for intra- (n=8) and inter-day (n=6) precisions at a concentration of 5 ng g-1 were 2.6 and 3.2%, respectively. The developed method has been successfully applied to 3-MCPD determination in refined edible oil samples including sunflower, corn, and canola oils.

Application of new N- and S-doped amorphous carbon in D-µSPE and its combination with deep eutectic solvent-based DLLME for the extraction of some mycotoxins from soymilk.

A new amorphous carbon-based dispersive micro solid-phase extraction method was developed for the extraction and preconcentration of several mycotoxins from soymilk samples. The extracted analytes were concentrated by a deep eutectic solvent-based dispersive liquid-liquid microextraction method, and then, quantified by a high-performance liquid chromatography-fluorescence detector. The sorbent was prepared from ß-cyclodextrin and methionine under mild conditions. The sorbent was doped by N and S, which improved its physicochemical properties. The optimization and validation of the method were performed using the "one-variable-at-a-time" method and International Council Harmonization guideline, respectively. Under the optimal conditions, low limits of detection and quantifications in the ranges of 0.08-0.56 and 0.27-1.9 ng L-1 were obtained, respectively. Also, intra- (n = 6) and inter-day (n = 6) precisions showed an acceptable repeatability of the present work as they were in the ranges of 3.9-6.2 and 4.6-8.9% at a concentration of 3 ng L-1 of each analyte, respectively. Finally, the proposed method was performed on different soymilk samples marketed in Tabriz city, and aflatoxin B1 was found in all samples. One soymilk was contaminated by ochratoxin A.

A minireview on nanoparticle-based sensors for the detection of coronaviruses.

Coronaviruses (CoVs) are a class of viruses that cause respiratory tract infections in birds and mammals. Severe acute respiratory syndrome and Middle East respiratory syndrome are pathogenic human viruses. The ongoing coronavirus causing a pandemic of COVID-19 is a recently identified virus from this group. The first step in the control of spreading the disease is to detect and quarantine infected subjects. Consequently, the introduction of rapid and reliable detection methods for CoVs is crucial. To date, several methods were reported for the detection of coronaviruses. Nanoparticles play an important role in detection tools, thanks to their high surface-to-volume ratio and exclusive optical property enables the development of susceptible analytical nanoparticle-based sensors. The studies performed on using nanoparticles-based (mainly gold) sensors to detect CoVs in two categories of optical and electrochemical were reviewed here. Details of each reported sensor and its relevant analytical parameters are carefully provided and explained.

Osteogenic Differentiation of Mesenchymal Stem Cells via Curcumin-Containing Nanoscaffolds.

The diverse pleiotropic pharmacological effects of curcumin nanoformulations have turned it into an attractive natural compound in different health-related problems. A great body of evidence has shown the impact of curcumin and its nanoformulations on the differentiation of stem cells. The current review highlights cellular and molecular mechanisms connected with the osteogenic differentiation of mesenchymal stem cells (MSCs) in the scaffolds benefiting from the presence of nanocurcumin pointing toward the role of inhibitory or stimulant signal transduction pathways in detail. Moreover, the effects of different concentrations as well as the structural modifications of curcumin on the differentiation of MSCs have been addressed.

A View on Polymerase Chain Reaction as an Outstanding Molecular Diagnostic Technique in Periodontology.

OBJECTIVES: This study presents a discussion on the fundamentals of polymerase chain reaction (PCR) and its use as a diagnostic tool in periodontology. MATERIALS AND METHODS: A computer-aided as well as hand-made search in PubMed and Scopus indexed journals (relevant to the topic) was done by keywords of molecular technique in periodontology, PCR, applications of PCR, and PCR in periodontics. Only the papers in the English language and outlining PCR and its association with periodontology were collected and utilized to provide a succinct review. There was no limitation for publication time. RESULTS: The results of our search showed that PCR has turned into a standard in diagnosis in the field of periodontology. A variety of researches has demonstrated that its sensitive, and specific characteristics make it a quick and effective technique of recognition, identification, and quantification of microorganisms. Identification of various immunoinflammatory markers at the mRNA expression level as well as ascertaining gene-related polymorphisms can also be performed. CONCLUSIONS: The mechanisms of periodontal disease can further become clarified using PCR. Clinical Relevance. PCR as a diagnostic method can play a main part in the validation of the clinical diagnosis of periodontal disease indicating the reason, pathogenesis, clinical steps, progress, and prognosis of the disease.

Direct quantitative detection of host cell residual DNA in recombinant Filgrastim by qPCR.

Host cell residual DNA is considered as an impurity in recombinant biopharmaceuticals. This study aimed to develop a direct qPCR method to quantify E. Coli residual DNA in recombinant Filgrastim. The specific primers were designed to amplify E. Coli's 16S-rDNA genomic region, which encodes the 16S-rRNA. The developed qPCR method showed that the designed primer has specifically amplified the target genome without any secondary reaction. The designed primer was also able to amplify the target gene as a representative of residual DNA in the drug matrix. Results show that the amount of residual DNA in Filgrastim is undetectable.

Combination of solvent extraction with deep eutectic solvent based dispersive liquid-liquid microextraction for the analysis of aflatoxin M1 in cheese samples using response surface methodology optimization.

A new extraction procedure based on combination of a solvent extraction and deep eutectic solvent based dispersive liquid-liquid microextraction has been introduced for the extraction of aflatoxin M1 from cheese samples. In this method, acetonitrile, deionized water, and n-hexane are added onto the sample and vortexed. Owning to different affinities of the substances in cheese toward the mentioned solvents, an efficient and selective extraction of the analyte is done in the acetonitrile phase. After centrifugation, the acetonitrile phase is removed and mixed with a new hydrophobic deep eutectic solvent prepared from N,N-diethanol ammonium chloride and carvacrol as an extraction solvent. The mixture is injected into deionized water, and a cloudy solution is obtained. Eventually, an aliquot of the organic phase is injected into high-performance liquid chromatography-fluorescence detection. After optimizing the effective parameters with the response surface methodology and a quadratic model, limits of detection and quantification were 0.74 and 2.56 ng/kg, respectively. The obtained extraction recovery and enrichment factor were 94% and 94, respectively. Also, intra- (n = 6) and interday (n = 4) precisions were less than or equal to 8.6% at a concentration of 5 ng/kg. The suggested method was applied to determine aflatoxin M1 in different cheese samples.

Preparation and characterization of a novel thermosensitive and bioadhesive cephalexin nanohydrogel: a promising platform for topical antibacterial delivery.

OBJECTIVE: Impetigo is a common and highly contagious bacterial skin infection that mostly affects young children and infants. Herein, we report the development of a thermosensitive and bioadhesive in-situ hydrogel-forming system containing cephalexin-loaded nanoparticles (NPs) suitable for antibacterial drug delivery. METHODS: The nanohydrogel was formulated using drug-loaded NPs and characterized by its physicochemical characteristics. Antibacterial activities of the cephalexin NPs and nanohydrogel were examined in vitro against Staphylococcus aureus (S. aureus). The ex vivo drug permeation study was performed using rat skin. Finally, this formulation was tested for in vivo antibacterial activity using superficial skin infections in rats. RESULTS: The mean size and entrapment efficiency of the NPs were found to be 178 nm and 58%, respectively. The transmission electron microscopy analysis verified the formation of spherical NPs. The drug-loaded NPs showed an enhanced eradication effect against S. aureus according to the declined MIC values in comparison with the untreated drug. The ex vivo permeation profile of the cephalexin nanohydrogel showed a slow release pattern for 8 h. When applied on rat skin for 6 days, the nanohydrogel exhibited a superior antibacterial activity with normal hair growth and skin appearance as compared to the plain drug hydrogel. CONCLUSIONS: These findings suggested that the nanohydrogel could serve as a valuable drug delivery platform against superficial bacterial infections.

Antimicrobial Activity of Nanostructured Lipid Carriers Loaded Punica granatum Seed Oil against Staphylococcus epidermidis.

BACKGROUND: Nanostructured lipid carriers (NLCs), due to their impressive benefits, have recently been considered in different areas. Besides, NLC loaded with essential oils is attractive for finding more effective antimicrobial products, especially against common bacteria such as Staphylococcus epidermidis (S. epidermidis). OBJECTIVE: This study aims to prepare and characterize NLCs encapsulated with Punica granatum (P. granatum) seed oil (PGS oil-loaded NLCs) and examine the antimicrobial effect of this combination against S. epidermidis. METHODS: PGS oil-loaded NLCs were prepared using a hot melt homogenization method. Later, they were characterized by determining particle size distribution (particle size analyzer), morphology (scanning electron microscopy (SEM)), and zeta potential (surface charge of NLCs). Minimum inhibitory concentrations (MIC) of PGS oil-loaded NLCs were assessed and compared with seed oil emulsion of P. granatum against S. epidermidis. RESULTS: PGS oil-loaded NLCs were spherical shaped nanoparticles, with a mean size of 102.10 nm and narrow size distribution (PDI = 0.26). The antibacterial assay showed PGS oil-loaded NLCs to have a higher in vitro antimicrobial activity than seed oil emulsion of P. granatum. CONCLUSION: To conclude, NLCs may be a favorable carrier to develop new antimicrobial agents. Lay Summary: The lipid nanoparticles such as nanostructured lipid carriers (NLCs) appeared as products first on the cosmetic market. Their advantages help them to be used in different healthcare and cosmetic products. With regard to previous studies, Punica granatum (P. granatum) extract shows antimicrobial and antioxidant properties that could be a valuable natural source against the wide ranges of bacteria. Then, P. granatum seed oil (PGS oil-loaded NLCs) prepared in this study can be used in dental and skin-related materials as a new natural antimicrobial product.

A brief overview on nano-sized materials used in the topical treatment of skin and soft tissue bacterial infections.

Introduction: Skin and soft tissue infections are a significant clinical problem that can happen anywhere on the body. Bacteria are the most common cause of skin and soft tissue infections in humans. Despite the fact that there is a lot of antimicrobial agents and antibiotics for elucidating bacterial infections, the prevention and control of infectious diseases continue to be one of the greatest challenges for public health worldwide. At the present time, an alarming increase in multidrug resistance instantly requests to find suitable alternatives to current antibiotics. Therefore, drug resistance has been attempted to be resolved by the development of new classes of antimicrobial agents or targeted delivery systems for antibacterial drugs using nanotechnology.Area covered: The present review summarizes the emerging topical efforts to support the use of nano-sized materials as a new opportunity to combat today's skin infectious diseases.Expert opinion: Nano-sized materials can overcome the stratum corneum barrier and deliver drugs specifically to bacterial skin infections with trivial side effects. Depending on the physicochemical characteristics of nano-scaled materials, they can specifically be selected to target bacterial pathogens and also to get into the skin layers. These systems can overcome the antibiotic-resistance mechanisms and help us to the design of novel topical formulations that will make administration of antibacterial compounds safer, easier and more convenient.

Freeze-thaw-induced cross-linked PVA/chitosan for oxytetracycline-loaded wound dressing: the experimental design and optimization.

Oxytetracycline is an antibiotic for the treatment of the infections caused by Gram-positive and Gram-negative microorganisms. Among novel formulations applied for damaged skin, hydrogels have shown to be superior as they can provide a moist environment for the wound. The purpose of this study was to prepare and evaluate the hydrogels of oxytetracycline consisted of polyvinyl alcohol (PVA) and chitosan polymers. A study design based on 4 factors and 3 levels was used for the preparation and evaluation of hydrogels formed by freeze-thaw (F-T) cycle using PVA and chitosan as a matrix-based wound dressing system. Furthermore, an experimental design was employed in order to study the effect of independent variables, namely drug amount (X1, 500-1000 mg), the amount of PVA (X2, 3.33-7.5%), the amount of chitosan (X3, 0.5-1%), and F-T cycle (X4, 3-7 cycles) on the dependent variables, including encapsulation efficiency, swelling index, adsorption of protein onto hydrogel surface, and skin permeation. The interaction of formulation variables had a significant effect on both physicochemical properties and permeation. Hydrogel microbial tests with sequential dilution method in Muller-Hinton broth medium were also carried out. The selected hydrogel (F6) containing 5% PVA, 0.75% chitosan, 1000 mg drug, and 3 F-T cycles was found to have increased encapsulation efficiency, gel strength, and higher skin permeation suitable for faster healing of wounds. Results showed the biological stability of oxytetracycline HCl in the hydrogel formulation with a lower dilution of the pure drug. Thus, oxytetracycline-loaded hydrogel could be a potential candidate to be used as a wound dressing system.

An update on calcium carbonate nanoparticles as cancer drug/gene delivery system.

INTRODUCTION: In recent years, the applications of calcium carbonate (CaCO3) nanoparticles (NPs) have gained extensive interest as targeted drug/gene delivery systems to cancerous tissues and cells due to their accessibility, low cost, safety, biocompatibility, pH-sensitivity, and slow biodegradability. AREAS COVERED: Drug-loaded CaCO3 NPs (CCNPs) have been reviewed. An updated search on the current state of CCNPs as cancer drug/gene delivery systems with a focus on their special properties including pH-sensitivity, biodegradability, and sustained release performance has been also assessed. EXPERT OPINION: Based on the reviewed literature, CCNPs, because of their superior features, will have a great aiding role in safe and efficient cancer treatment in the near future.

Evaluation of Herbal Mouthwashes Containing Zataria Multiflora Boiss, Frankincense and Combination Therapy on Patients with Gingivitis: A Double-Blind, Randomized, Controlled, Clinical Trial.

BACKGROUND: Dental plaques as adhesive microbial aggregates on tooth surfaces are considered the first stage of tooth decay as well as gingivitis. Accordingly, the effect of different antimicrobial mouthwashes on removing dental plaques and preventing their formation has been evaluated in various studies. This study aimed to evaluate the efficacy of herbal mouthwashes containing hydro-alcoholic extract of Zataria multiflora (ZM), Frankincense (FR), and a combination of both (ZM+FR) and compare it with chlorhexidine (CHX) mouthwash in subjects with gingivitis. MATERIALS AND METHODS: In this randomized, controlled, clinical trial a total of 140 patients with gingivitis were divided into four groups including CHX (control group), ZM, FR, and ZM+FR groups. Plaque index (PI), gingival index (GI), and gingival bleeding index (GBI) were measured in days 1, 14, and 21. RESULTS: All three herbal types of mouthwash significantly improved plaque, gingivitis, and gingival bleeding throughout days 14 to 21 (P<0.001). There was no difference between herbal mouthwash with CHX groups. CHX mouthwash showed the most side effects (54.3%), while ZM mouthwash showed the least side effects and the highest consumer satisfaction (5.7% and 94%, respectively). CONCLUSION: All of the herbal mouthwashes can be good candidates for controlling gingivitis. Comparing with CHX mouthwash, herbal mouthwashes have lower side effects and negligible alcohol content. Among the herbal mouthwashes, ZM outperforms FR and FR+ZM due to its lower side effects and higher levels of patients' satisfaction.