Efficacy of hypochlorous acid as an alternative oral antimicrobial agent on human gingival fibroblasts, Aggregatibacter actinomycetemcomitans, and Candida albicans biofilms in vitro.

This study compared the cytotoxicity and antimicrobial activity of hypochlorous acid (HOCl) at 50 ppm and 200 ppm and 0.2% chlorhexidine (CHX) at various time intervals, in vitro. Cell viability and cytotoxicity of human gingival fibroblasts (HGF) were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and the lactate dehydrogenase assay. Antimicrobial effects on Aggregatibacter actinomycetemcomitans and Candida albicans were determined using the time-kill method. All solutions exhibited a significant impact on HGFs in a dose- and time-dependent manner. 50 ppm HOCl demonstrated the highest cell viability, followed by 200 ppm HOCl. Both HOCl solutions were less cytotoxic to HGFs than 0.2% CHX. 50 ppm and 200 ppm HOCl demonstrated stronger efficiencies than CHX against A. actinomycetemcomitans and C. albicans. The data suggest that HOCl solutions have potential as an alternative antiseptic to CHX due to their lower cytotoxicity and superior antimicrobial activity, but optimal dosage of HOCl requires further investigations.

Clinical assessment of various imaging systems for dental plaque scoring after the use of 3 different toothpastes.

Purpose: This study was conducted to compare dental plaque scores obtained through clinical examinations and various imaging techniques, as well as to assess the effectiveness of herbal and conventional toothpastes for plaque removal. Material and Methods: Thirty volunteers were divided into 3 groups. Each group was given a different toothpaste (from 2 herbal toothpastes and a conventional toothpaste) with which to brush their teeth for 21 days. Both initially and after brushing, dental plaque samples were collected, and plaque on the buccal surfaces of anterior teeth was scored using several imaging systems after staining with a disclosing agent. Specifically, digital dental photography, intraoral digital scanning, and FluoreCam imaging were employed to capture intraoral images. The Turesky Modified Quigley-Hein Plaque Index was used for clinical examination and image analysis. Quantitative polymerase chain reaction analyses and correlational assessments between clinical examination and imaging scores were conducted before and after toothpaste use. The Shapiro-Wilk test and Pearson correlations were utilized. Results: The lowest mean value was observed in the clinical examination without staining, while the highest was obtained using the FluoreCam method. No significant change was found in the level of any microorganism assessed following toothpaste use (P<0.05), with the exception of a decrease in S. mutans levels after using conventional toothpaste (P<0.05). Conclusion: Herbal toothpaste demonstrated plaque-removal effectiveness comparable to that of conventional toothpaste. The use of imaging methods for measuring plaque index has been suggested as a means to educate patients about plaque control and promote ongoing oral care.

Antibiotic resistance in Campylobacter jejuni and Campylobacter coli: significant contribution of an RND type efflux pump in erythromycin resistance.

In this study, we aimed to determine the antibiotic resistance status of Campylobacter spp. isolated from human infections in our region, including the role of mechanisms involved in erythromycin resistance. Standard methods were used for the isolation, identification and antibiotic susceptibility testing of Campylobacter spp. isolates. Erythromycin-resistant mutants were selected from erythromycin-susceptible clinical isolates, and the erythromycin resistance mechanisms were investigated phenotypically by determining the erythromycin MICs of isolates in the presence and absence of the resistance nodulation cell division (RND) type efflux pump inhibitor, phenylalanine-arginine ß-naphthylamide dihydrochloride (PAßN), and genotypically by determining ribosomal and cmeABC alterations using PCR and DNA sequence analysis. Campylobacter spp., including 184 C. jejuni and 20 C. coli in a two-year period, were the most frequently isolated gastrointestinal bacterial pathogens in our region. However, in both C. jejuni and C. coli, resistance to tetracycline and ciprofloxacin were found to be high, erythromycin resistance was especially high (20%) in C. coli. With a ribosomal alteration, A2075G, which was found to be associated with high-level erythromycin resistance in clinical isolates, PAßN significantly reduced the erythromycin MICs in both clinical isolates and mutants. An important finding of this study, while considering cmeABC operon, is the explanation of why erythromycin resistance is more common among C. coli than C. jejuni, bearing in mind the specific deletions and alterations in the intergenic region of the operon in all erythromycin-resistant C. coli isolates. Ultimately, these findings revealed the significant role of RND-type efflux activity in increased erythromycin MICs of the isolates.

Fucoidan-loaded electrospun Polyvinyl-alcohol/Chitosan nanofibers with enhanced antibacterial activity for skin tissue engineering.

The polymeric nanofiber may interact and control certain regeneration processes at the molecular level to repair damaged tissues. This research focuses on the development of characterization and antibacterial capabilities of polyvinyl alcohol (PVA)/chitosan (CS) nanofibres containing fucoidan (FUC) for tissue engineering as a skin tissue substitute. A control group consisting of 13% PVA/(0.1)% CS nanofiber was prepared. To confer antibacterial properties to the nanofiber, 10, 20, and 30 mg of FUC were incorporated into this control group. The scanning electron microscope (SEM) proved the homogeneous and beadless structures of the nanofibers. The antibacterial activity of the 13% PVA/(0.1)% CS/(10, 20, 30) FUC was tested against the S.aureus and E.coli and the results showed that with FUC addition, the antibacterial activities of the nanofibers increased. The biocompatibility test was performed with a fibroblast cell line for 1, 3, and 7 days of incubation and the results demonstrated that FUC addition enhanced the bioactivity of the 13% PVA/(0.1)% CS nanofibers. In addition, the biocompatibility results showed that 13% PVA/(0.1)% CS/10 FUC had the highest viability value for all incubation periods compared to the others. In addition, the tensile test results showed that; the maximum tensile strength value was observed for 13% PVA/(0.1)% CS/10 FUC nanofibers.

Development of gallic acid-loaded ethylcellulose fibers as a potential wound dressing material.

In this study, novel fibers were designed based on ethylcellulose (EC), loaded with different concentrations of gallic acid (GA) using the electrospinning technique, in order to investigate the potential of these materials as wound dressings. The chemical structure and morphology, along with the antimicrobial and biocompatibility tests of the EC_GA fibers were investigated. To observe the chemical interactions between the components, fourier transform infrared spectroscopy (FTIR) was used. The morphological analyzes were performed using scanning electron microscope (SEM). The uniaxial tensile test machine was used to obtain mechanical performance of the fibers. MTT assay was applied to get the biocompatibility properties of the fibers and antimicrobial test was applied to obtain the antimicrobial activity of the fibers. Based on the obtained results, the highest viability value of 67.4 % was obtained for 10%EC_100GA on the third day of incubation, demonstrating that with the addition of a higher concentration of GA, the cell viability increases. The antimicrobial tests, evaluated against Staphylococcus (S.) aureus, Escherichia (E.) coli, Pseudomonas (Ps.) aeruginosa and Candida (C.) albicans, showed a >90 % microbial reduction capacity correlated with a logarithmic reduction ranging from 0.63 to 1, for 10%EC_100 GA. In vitro release tests of GA from the fibers showed that GA was totally released from 10%EC_100 GA fibers after 2880 min, demonstrating a controlled release profile. These findings demonstrated that EC_GA fibers may be suitable for application in biomedical fields such as wound dressing materials. However, further studies should be performed to increase the biocompatibility properties of the fibers.

Biocompatible polyvinyl alcohol nanofibers loaded with amoxicillin and salicylic acid to prevent wound infections.

Diabetic wounds are one of the most challenging clinical conditions in diabetes, necessitating the development of new treatments to foster healing and prevent microbial contamination. In this study, polyvinyl alcohol was used as a matrix polymer, and amoxicillin (AMX) and salicylic acid (SA) were selected as bioactive compounds with antimicrobial (with AMX) and anti-inflammatory action (with SA) to obtain innovative drug-loaded electrospun nanofiber patches for the management of diabetic wounds. Scanning electron microscope images revealed the uniform and beadless structure of the nanofiber patches. Mechanical tests indicated that AMX minimally increased the tensile strength, while SA significantly reduced it. The patches demonstrated effective antibacterial activity against both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) strains. The potential of these patches in the development of novel wound dressings is highlighted by the excellent biocompatibility with fibroblast cells maintained for up to 7 d.

A Novel Approach for the Fabrication of 3D-Printed Dental Membrane Scaffolds including Antimicrobial Pomegranate Extract.

In this study, a dental membrane scaffold was fabricated using a 3D printing technique, and the antimicrobial effect of pomegranate seed and peel extract were investigated. For the production of the dental membrane scaffold, a combination of polyvinyl alcohol, starch, and pomegranate seed and peel extracts was used. The aim of the scaffold was to cover the damaged area and aid in the healing process. This can be achieved due to the high antimicrobial and antioxidant content of pomegranate seed and peel extracts (PPE: PSE). Moreover, the addition of starch and PPE: PSE improved the biocompatibility of the scaffold, and their biocompatibility was tested using human gingival fibroblast (HGF) cells. The addition of PPE: PSE into the scaffolds resulted in a significant antimicrobial effect on S. aureus and E. faecalis bacteria. Moreover, different concentrations of starch (1%, 2%, 3% w/v) and pomegranate peel and seed extract (3%, 5%, 7%, 9%, and 11% PE v/v) were analyzed to obtain the ideal dental membrane structure. The optimum starch concentration was chosen as 2% w/v due to it giving the scaffold the highest mechanical tensile strength (23.8607 ± 4.0796 MPa). The pore sizes of each scaffold were studied by SEM analysis, and pore sizes were arranged between 155.86 and 280.96 µm without any plugging problems. Pomegranate seed and peel extracts were obtained by applying the standard extraction method. High-performance liquid chromatography was performed using the diode-array detection (HPLC-DAD) technique to analyze the phenolic content of the pomegranate seed and peel extracts. Two phenolic components of the pomegranate seed and peel extracts were investigated in the following amounts: fumaric acid (17.56 µg analyte/mg extract) and quinic acid (18.79 µg analyte/mg extract) in pomegranate seed extract and fumaric acid (26.95 µg analyte/mg extract) and quinic acid (33.79 µg analyte/mg extract) in pomegranate peel extract.

3D-Printed PCL Scaffolds Combined with Juglone for Skin Tissue Engineering.

Skin diseases are commonly treated with antihistamines, antibiotics, laser therapy, topical medications, local vitamins, or steroids. Since conventional treatments for wound healing (skin allografts, amnion, xenografts, etc.) have disadvantages such as antigenicity of the donor tissue, risk of infection, or lack of basement membrane, skin tissue engineering has become a popular new approach. The current study presents the design and fabrication of a new wound-dressing material by the addition of Juglone (5-hydroxy-1,4-naphthoquinone) to a 25% Polycaprolactone (PCL) scaffold. Juglone (J) is a significant allelochemical found in walnut trees and, in this study is used as a bioactive material. The effects of different amounts of J (1.25, 2.5, 5, 7.5, and 10 mg) on the biocompatibility, mechanical, chemical, thermal, morphological, and antimicrobial properties of the 3D-printed 25% PCL scaffolds were investigated. The addition of J increased the pore diameter of the 25% PCL scaffold. The maximum pore size (290.72 ± 14 µm) was observed for the highest amount of J (10 mg). The biocompatibility tests on the scaffolds demonstrated biocompatible behavior from the first day of incubation, the 25% PCL/7.5 J scaffold having the highest viability value (118%) among all of the J-loaded scaffolds. Drug release of J into phosphate buffered saline (PBS) at pH 7.4 showed that J was completely released from all 25% PCL/J scaffolds within 7 days of incubation.

Fabrication of Electrospun Juglans regia (Juglone) Loaded Poly(lactic acid) Scaffolds as a Potential Wound Dressing Material.

Juglone (5-hydroxy-1,4-naphthoquinone) (J) is a naphthoquinone structured allelochemical that is mostly found in the roots, leaves, nut-hulls, bark, and wood of walnut (Juglans regia). In this study, the biocompatibility, mechanical, thermal, chemical, morphological, and antimicrobial properties of the poly(lactic acid) (PLA) (w/v)/J (10, 20, 30 mg) electrospun scaffolds were investigated. Based on the results of the study, it was shown that juglone addition increased the antimicrobial properties of the scaffolds against the Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli), compared to the neat PLA film after 24 h of contact time. According to the tensile test results, the addition of J made the scaffolds more flexible but decreased the mechanical strength. The cytotoxicity properties of the J-added scaffolds demonstrated a toxic behavior on the first day of incubation. However, with an increase in the J ratio, the fibroblast cell metabolic activity increased for all incubation periods.

Chondromyxoid Fibroma of the Calcaneus: A Rare Case Report.

Chondromyxoid fibroma (CMF) of the calcaneus is extremely rare. We report a case of CMF of the calcaneus in a 34-year-old female. She had foot pain for one year and had increased pain for the last two months. The patient complained of limping due to the pain she felt. CMF of the calcaneus was treated with curettage and bone grafting. The patient was allowed to mobilize the very next day of surgery with weight bearing as much as she could tolerate. No recurrence was encountered during the 18-month follow-up of the patient. The patient could perform activities in her daily life painlessly. Carefully performed curettage and bone grafting is an effective treatment method in the treatment of CMF of the calcaneus. CMF in the calcaneus may not be as rare as it is thought, and should be considered in the differential diagnosis.

Diversity of HIV-1 Subtypes and Transmitted Drug-resistance Mutations Among Minority HIV-1 Variants in a Turkish Cohort.

BACKGROUND: The World Health Organization (WHO) recommends the surveillance of transmitted drug resistance mutations (TDRMs) to ensure the effectiveness and sustainability of HIV treatment programs. OBJECTIVE: Our aim was to determine the TDRMs and evaluate the distribution of HIV-1 subtypes using and compared next-generation sequencing (NGS) and Sanger-based sequencing (SBS) in a cohort of 44 antiretroviral treatment-naïve patients. METHODS: All samples that were referred to the microbiology laboratory for HIV drug resistance analysis between December 2016 and February 2018 were included in the study. After exclusions, 44 treatment-naive adult patients with a viral load of >1000 copies/mL were analyzed. DNA sequencing for reverse transcriptase and protease regions was performed using both DeepChek ABL single round kit and Sanger-based ViroSeq HIV-1 Genotyping System. The mutations and HIV-1 subtypes were analyzed using the Stanford HIVdb version 8.6.1 Genotypic Resistance software, and TDRMs were assessed using the WHO surveillance drug-resistance mutation database. HIV-1 subtypes were confirmed by constructing a maximum-likelihood phylogenetic tree using Los Alamos IQ-Tree software. RESULTS: NGS identified nucleos(t)ide reverse transcriptase inhibitor (NRTI)-TDRMs in 9.1 % of the patients, non-nucleos(t)ide reverse transcriptase inhibitor (NNRTI)-TDRMs in 6.8 % of the patients, and protease inhibitor (PI)-TDRMs in 18.2 % of the patients at a detection threshold of ≥ 1 %. Using SBS, 2.3 % and 6.8 % of the patients were found to have NRTI- and NNRTI-TDRMs, respectively, but no major PI mutations were detected. M41L, L74I, K65R, M184V, and M184I related to NRTI, K103N to NNRTI, and N83D, M46I, I84V, V82A, L24I, L90M, I54V to the PI sites were identified using NGS. Most mutations were found in low-abundance (frequency range: 1.0 % - 4.7 %) HIV-1 variants, except M41L and K103N. The subtypes of the isolates were found as follows; 61.4 % subtype B, 18.2 % subtype B/CRF02_AG recombinant, 13.6 % subtype A, 4.5 % CRF43_ 02G, and 2.3 % CRF02_AG. All TDRMs, except K65R, were detected in HIV-1 subtype B isolates. CONCLUSION: The high diversity of protease site TDRMs in the minority HIV-1 variants and prevalence of CRFs were remarkable in this study. All minority HIV-1 variants were missed by conventional sequencing. TDRM prevalence among minority variants appears to be decreasing over time at our center.

Propolis-Based Nanofiber Patches to Repair Corneal Microbial Keratitis.

In this research, polyvinyl-alcohol (PVA)/gelatin (GEL)/propolis (Ps) biocompatible nanofiber patches were fabricated via electrospinning technique. The controlled release of Propolis, surface wettability behaviors, antimicrobial activities against the S. aureus and P. aeruginosa, and biocompatibility properties with the mesenchymal stem cells (MSCs) were investigated in detail. By adding 0.5, 1, and 3 wt.% GEL into the 13 wt.% PVA, the morphological and mechanical results suggested that 13 wt.% PVA/0.5 wt.% GEL patch can be an ideal matrix for 3 and 5 wt.% propolis addition. Morphological results revealed that the diameters of the electrospun nanofiber patches were increased with GEL (from 290 nm to 400 nm) and Ps addition and crosslinking process cause the formation of thicker nanofibers. The tensile strength and elongation at break enhancement were also determined for 13 wt.% PVA/0.5 wt.% GEL/3 wt.% Ps patch. Propolis was released quickly in the first hour and arrived at a plateau. Cell culture and contact angle results confirmed that the 3 wt.% addition of propolis reinforced mesenchymal stem cell proliferation and wettability properties of the patches. The antimicrobial activity demonstrated that propolis loaded patches had antibacterial activity against the S. aureus, but for P. aeruginosa, more studies should be performed.

3D Propolis-Sodium Alginate Scaffolds: Influence on Structural Parameters, Release Mechanisms, Cell Cytotoxicity and Antibacterial Activity.

In this study, the main aim was to fabricate propolis (Ps)-containing wound dressing patches using 3D printing technology. Different combinations and structures of propolis (Ps)-incorporated sodium alginate (SA) scaffolds were developed. The morphological studies showed that the porosity of developed scaffolds was optimized when 20% (v/v) of Ps was added to the solution. The pore sizes decreased by increasing Ps concentration up to a certain level due to its adhesive properties. The mechanical, swelling-degradation (weight loss) behaviors, and Ps release kinetics were highlighted for the scaffold stability. An antimicrobial assay was employed to test and screen antimicrobial behavior of Ps against Escherichia coli and Staphylococcus aureus strains. The results show that the Ps-added scaffolds have an excellent antibacterial activity because of Ps compounds. An in vitro cytotoxicity test was also applied on the scaffold by using the extract method on the human dermal fibroblasts (HFFF2) cell line. The 3D-printed SA-Ps scaffolds are very useful structures for wound dressing applications.

An alternative for urine cultures: Direct identification of uropathogens from urine by MALDI-TOF MS.

Urinary tract infections are one of the most common bacterial infections and rapid diagnosis of the infection is essential for appropriate antibiotic therapy. The goal of our study was to identify urinary pathogens directly by MALDI-TOF MS and to perform antibiotic susceptibility tests in order to shorten the period spent for culturing.Urine samples submitted for culture to the Clinical Microbiology Laboratory were enrolled in this study. Urine samples were screened for leukocyte and bacteria amount by flow cytometry. Samples with bacterial load of 106-107/mL were tested directly by MALDI-TOF MS and antibiotic susceptibility tests (AST) were performed.In total, 538 positive urine samples were evaluated in our study. MALDI-TOF MS identified the microorganism directly from the urine sample in 91.8% of these samples and the concordance rate of conventional identification and direct detection was 95.8% for Gram-negatives at the genus and species level. Escherichia coli (n:401) was the most frequently isolated microorganism, followed by Klebsiella pneumoniae (n:57). AST results were generated for 111 of these urine samples and the concordance was 90% and 87% for E. coli and K. pneumoniae, respectively.Our results showed that screening of urine samples with flow cytometry to detect positive samples and identification of uropathogens directly by MALDI-TOF MS with an accuracy of over 90% can be a suitable method particularly for Gram-negative bacteria in clinical microbiology laboratories.

Development of Satureja cuneifolia-loaded sodium alginate/polyethylene glycol scaffolds produced by 3D-printing technology as a diabetic wound dressing material.

Acute wounds are a common health problem, with millions of people affected and decreased granulation tissue formation and vascularization, it is also a big challenge for wound care researchers to promote acute wound healing around the globe. This study aims to produce and characterize Satureja cuneifolia plant extract (SC)-blended with sodium alginate (SA) /polyethylene glycol (PEG) scaffolds for the potential treatment of diabetic ulcer. SA/PEG scaffolds were prepared by adding different concentrations (1, 3, and 5 wt%) of PEG to 9 wt% SA. The morphological and chemical composition of the resulting 3D printed composite scaffolds was determined using scanning electron microscopy (SEM) and Fourier transforms infrared spectroscopy (FTIR), respectively. Mechanical and thermal properties, swelling, and degradation behaviours were also investigated. The release kinetics of SC were performed. The antimicrobial analysis was evaluated against Escherichia coli and Staphylococcus aureus strains. 3D printed scaffolds have shown an excellent antibacterial effect, especially against gram-positive bacteria due to the antibacterial SC extract they contain. Furthermore, the cell viability of fibroblast (L929) cells on/within scaffolds were determined by the colourimetric MTT assay. The SA/PEG/SC scaffolds show a great promising potential candidate for diabetic wound healing and against bacterial infections.

Comparative in vitro activity of plazomicin and older aminoglyosides against Enterobacterales isolates; prevalence of aminoglycoside modifying enzymes and 16S rRNA methyltransferases.

Comparative in vitro activity of plazomicin and 4 older aminoglycosides was evaluated with broth microdilution in 714 blood isolates from 14 hospitals in Turkey. Isolates included Escherichia coli (n=320), Klebsiella spp. (n=294), Enterobacter spp. (n=69), Serratia marcescens (n=20), and Citrobacter spp. (n=11). Isolates resistant to older aminoglycosides (n=240) were screened for aminoglycoside modifying enzyme genes: aac(6')-Ib, aac(3)-Ia, aac(3)-IIa, ant(2″)-Ia. Isolates with high MICs for plazomicin (n=41) were screened for 16S rRNA methyltransferase genes (armA, rmtA, rmtB, rmtC, rmtD, rmtE, rmtF, rmtG, rmtH, npmA) and 2 carbapenemase genes (blaOXA-48, blaNDM-1). Overall, resistance to plazomicin, amikacin, netilmicin, gentamicin, and tobramycin was 7.7%, 7.4%, 31.5%, 32.9%, and 34.7%, respectively. aac(6')-Ib and aac(3)-IIa were the most common AME genes. Co-occurrence of blaNDM-1 with armA and rmtC and blaOXA-48 with armA was striking. Enterobacter cloacae carrying rmtC+blaNDM-1, S. marcescens with armA+blaOXA-48, and rmtF+ blaOXA-48 in K. pneumoniae were reported for the first time.

Evaluation of Cytotoxicity and Mutagenicity of Wastewater from Istanbul: Data from Hospitals and Advanced Wastewater Treatment Plant.

Wastewater (WW) carry considerable amount of chemicals that could have mutagenic or cytotoxic effect from hospital discharges to aquatic environment. Our objective was to determinate the possible mutagenic and toxic effects of hospital originated WWs and effectiveness of the wastewater treatment plants (WTP) functions. In the study the mutagenic and cytotoxic potential of three hospitals and influent/effluent of a treatment plant WW collected in Istanbul and was examined using AMES, XTT, and lactate dehydrogenase (LDH) assays. Mutagenic effects were detected at both hospital discharges and advanced biological wastewater plant. We observed no cytotoxic effect in fibroblasts for LDH and XTT assays whereas high cytotoxicity for all samples was found in hepatocytes by XTT assay. According to the results even if advanced technology is used for treatment of WW, mutagenic and cytotoxic effects still remain, and the present technologies need to be further improved.