Antimicrobial Behavior and Cytotoxicity of Indocyanine Green in Combination with Visible Light and Water-Filtered Infrared A Radiation against Periodontal Bacteria and Subgingival Biofilm.

The widespread increase of antibiotic resistance highlights the need for alternative treatments such as antimicrobial photodynamic therapy (aPDT). This study aimed to evaluate the antimicrobial behavior and cytotoxicity of aPDT with indocyanine green (ICG) in combination with visible light (Vis) and water-filtered infrared A (wIRA). Representative periodontal bacteria (Parvimonas micra, Atopobium riame, Slackia exigua, Actinomyces naeslundii, Porphyromonas gingivalis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Prevotella nigrescens) and subgingival in situ biofilms from periodontal patients were treated with aPDT for 5 min. ICG was used at different concentrations (50-500 µg/mL) and the number of viable cells was determined in colony forming units (CFU). Untreated negative controls and 0.2% chlorhexidine as a positive control were also prepared. The cytotoxicity test on human keratinocytes in vitro was analyzed with the AlamarBlue assay after 5, 10, and 20 min, with four ICG concentrations, and at two temperatures (room temperature and 37 °C). The tested periodontal pathogens treated with aPDT were eliminated in a range between 1.2 and 6.7 log10 CFU, except for A. naeslundii, which was killed at a lower range. The subgingival biofilm treated with aPDT expressed significant differences to the untreated controls except for at 300 µg/mL ICG concentration. The cytotoxicity was directly related to the concentration of ICG and irradiation time. These observations raise questions concerning the use of this specific aPDT as an adjuvant to periodontal treatments due to its possible toxicity towards human gingival cells.

How Do Polymer Coatings Affect the Growth and Bacterial Population of a Biofilm Formed by Total Human Salivary Bacteria?-A Study by 16S-RNA Sequencing.

Antimicrobial surface modifications are required to prevent biomaterial-associated biofilm infections, which are also a major concern for oral implants. The aim of this study was to evaluate the influence of three different coatings on the biofilm formed by human saliva. Biofilms grown from human saliva on three different bioactive poly(oxanorbornene)-based polymer coatings (the protein-repellent PSB: poly(oxanorbornene)-based poly(sulfobetaine), the protein-repellent and antimicrobial PZI: poly(carboxyzwitterion), and the mildly antimicrobial and protein-adhesive SMAMP: synthetic mimics of antimicrobial peptides) were analyzed and compared with the microbial composition of saliva, biofilms grown on uncoated substrates, and biofilms grown in the presence of chlorhexidine digluconate. It was found that the polymer coatings significantly reduced the amount of adherent bacteria and strongly altered the microbial composition, as analyzed by 16S RNA sequencing. This may hold relevance for maintaining oral health and the outcome of oral implants due to the existing synergism between the host and the oral microbiome. Especially the reduction of some bacterial species that are associated with poor oral health such as Tannerella forsythia and Fusobacterium nucleatum (observed for PSB and SMAMP), and Prevotella denticola (observed for all coatings) may positively modulate the oral biofilm, including in situ.

Fine-mapping of angular leaf spot resistance gene Phg-2 in common bean and development of molecular breeding tools.

KEY MESSAGE: The Common Bean Angular Leaf Spot Resistance Gene Phg-2 was fine-mapped to a 409-Kbp region, and molecular markers for breeders were developed and validated in field experiments. Common bean (Phaseolus vulgaris L.) is an important food legume in Latin America, Asia and Africa. It is an important source of protein, carbohydrates and micro-minerals, particularly for smallholder farmers. Common bean productivity is affected by angular leaf spot (ALS) disease caused by the pathogenic fungus Pseudocercospora griseola, resulting in significant yield losses, particularly in low-input smallholder farming systems in the tropics. The ALS resistance gene Phg-2, which was found in several highly resistant common bean genotypes, was investigated in crosses between Mesoamerican pre-breeding lines and elite Andean breeding lines. Next-generation sequencing (NGS) data sets were used to design new SNP-based molecular markers. The Phg-2 locus was confirmed to be the major locus providing ALS resistance in these crosses. The locus was fine-mapped to a 409-Kbp region on chromosome 8. Two clusters of highly related LRR genes were identified in this region, which are the best candidate genes for Phg-2. Molecular markers were identified that are closely linked to the Phg-2 resistance gene and also highly specific to the donor germplasm. Marker-assisted selection (MAS) was used to introgress the Phg-2 resistance locus into Andean breeding germplasm using MAB lines. The usefulness of molecular markers in MAS was confirmed in several field evaluations in complex breeding crosses, under inoculation with different ALS pathotypes. This project demonstrates that NGS data are a powerful tool for the characterization of genetic loci and can be applied in the development of breeding tools.

A Simultaneously Antimicrobial, Protein-Repellent, and Cell-Compatible Polyzwitterion Network.

A simultaneously antimicrobial, protein-repellent, and cell-compatible surface-attached polymer network is reported, which reduces the growth of bacterial biofilms on surfaces through its multifunctionality. The coating was made from a poly(oxonorbornene)-based zwitterion (PZI), which was surface-attached and cross-linked in one step by simultaneous UV-activated CH insertion and thiol-ene reaction. The process was applicable to both laboratory surfaces like silicon, glass, and gold and real-life surfaces like polyurethane foam wound dressings. The chemical structure and physical properties of the PZI surface and the two reference surfaces SMAMP ("synthetic mimic of an antimicrobial peptide"), an antimicrobial but protein-adhesive polymer coating, and PSB (poly(sulfobetaine)), a protein-repellent but not antimicrobial polyzwitterion coating were characterized by Fourier transform infrared spectroscopy, ellipsometry, contact angle measurements, photoelectron spectroscopy, swellability measurements (using surface plasmon resonance spectroscopy, SPR), zeta potential measurements, and atomic force microscopy. The time-dependent antimicrobial activity assay (time-kill assay) confirmed the high antimicrobial activity of the PZI; SPR was used to demonstrate that it was also highly protein-repellent. Biofilm formation studies showed that the material effectively reduced the growth of Escherichia coli and Staphylococcus aureus biofilms. Additionally, it was shown that the PZI was highly compatible with immortalized human mucosal gingiva keratinocytes and human red blood cells using the Alamar Blue assay, the live-dead stain, and the hemolysis assay. PZI thus may be an attractive coating for biomedical applications, particularly for the fight against bacterial biofilms on medical devices and in other applications.

Development of a standardized and safe airborne antibacterial assay, and its evaluation on antibacterial biomimetic model surfaces.

Bacterial infection of biomaterials is a major concern in medicine, and different kinds of antimicrobial biomaterial have been developed to deal with this problem. To test the antimicrobial performance of these biomaterials, the airborne bacterial assay is used, which involves the formation of biohazardous bacterial aerosols. We here describe a new experimental set-up which allows safe handling of such pathogenic aerosols, and standardizes critical parameters of this otherwise intractable and strongly user-dependent assay. With this new method, reproducible, thorough antimicrobial data (number of colony forming units and live-dead-stain) was obtained. Poly(oxonorbornene)-based Synthetic Mimics of Antimicrobial Peptides (SMAMPs) were used as antimicrobial test samples. The assay was able to differentiate even between subtle sample differences, such as different sample thicknesses. With this new set-up, the airborne bacterial assay was thus established as a useful, reliable, and realistic experimental method to simulate the contamination of biomaterials with bacteria, for example in an intraoperative setting.

Morfología dental en dentición mixta: correlación de tres rasgos morfológicos en dientes temporales y permanentes de escolares de Cali, Colombia / Mixed dentition Morphology characteristics: a correlation between 3 non-metric dental traits in temporal and permanent dentition in scholars from Cali - Colombia

Objetivo: Determinar la correlación entre la frecuencia y la variabilidad de tres rasgos morfológicos dentales coronales (cúspide de Carabelli, protostilido y patrón cúspideo) entre los dientes temporales (um2/ lm2) y permanentes (UM1/LM1), con el fin de establecer la posible correlación de con su expresión o manifestación clínica. Materiales y Métodos: En modelos de yeso obtenidos de impresiones tomadas a 100 escolares de ambos sexos, estudiantes de tres colegios de Cali-Colombia, que presentaban dentición mixta con primeros molares permanentes y segundos molares temporales presentes en boca y pertenecían a un grupo de población mestiza, se realizó un estudio descriptivo cuantitativo para caracterizar la morfología dental mediante el sistema de placas ASU y los Métodos de Grine y de Sciulli. Resultados: La alta prevalencia de las formas del surco y de la fosa de la Cúspide de Carabelli, la alta frecuencia del protostílido en grado 1 (foramen secum) y el comportamiento de la expresión del patrón cuspídeo, evidenciaron que la población objeto de estudio ha recibido influencia de los complejos dentales mongoloide y caucasoide. Se hizo indudable la correspondencia en las formas de manifestación en ambas denticiones, y se observó simetría bilateral y que no se presenta dimorfismo sexual en los rasgos morfológicos dentales estudiados.

Objectives: Determine the correlation of the frequency and variability of three Non-metric dental crown traits (Carabellicusp, protostylid and molar pattern cusp) between deciduous (um2/lm2) and permanent (UM1/LM1) teeth in mixed-dentition subjects. Materials and methods: A descriptive study was conducted to determine the dental morphology in mixed dentition stage of young subjects. The Arizona State University Dental Anthropology System ASUDAS and Grind and Sciulli Methods were used as reference to compare frequency of dental traits in dental casts of 100 young individuals racially mixed. Results: A high expression of furrows and pits of the Carabelli cusp, a high frequency of protostylid in degree 1 (foramen secum) and a expression of molar pattern cusp, revealed that the studied population received influences of the Mongoloid and Caucasoid dental complexes. Correspondence on the traits expression among temporal and permanent dentition was also confirmed. Bilateral symmetry and sexual dimorphism in the analyzed non-metric dental crown traits were found.