Biofilm removal from Difficult-to-Reach places via secondary cavitation within a constrained geometry mimicking a Periodontal/Peri-Implant pocket.

Biofilm removal from the apical region of the periodontal or peri-implant pocket, which is very difficult to achieve with mechanical instruments, is a major unresolved issue in dentistry. Here, we propose the use of photoacoustically induced streaming and secondary cavitation to achieve superior cleaning efficacy in the apical region of the periodontal and peri-implant pocket. We have used a prefabricated narrow wedge system that mimics the consistency of periodontal and peri-implant pockets of both healthy and severely inflamed tissue. We studied the effect of single-pulse modality Er:YAG on Pseudomonas aeruginosa biofilm removal. We used different laser energies, fiber-tip positions, and laser treatment durations. The cleaning process was monitored in real-time with a high-speed camera after each individual laser pulse application. The obtained results suggest that biofilm cleaning efficacy in a difficult-to-reach place in healthy model tissue is directly related to the onset of secondary cavitation bubble formation, which correlates with a significant improvement of biofilm removal from the apical region of the periodontal or peri-implant pocket. In comparison to the healthy tissue model, the laser energy in inflamed tissue model had to be increased to obtain comparable biofilm cleaning efficacy. The advantage of photoacoustic cavitation compared to other methods is that laser-induced cavitation can trigger secondary cavitation at large distances from the point of laser application, which in principle allows biofilm removal at distant locations not reachable with a laser fiber tip or other mechanical instruments.

Molecular characterization of extended-spectrum ß-lactamase-producing Escherichia coli isolated from lower respiratory tract samples between 2002 and 2019 in the Central Slovenia region.

BACKGROUND: Antibiotic resistance is one of the most serious global health problems and threatens the effective treatment of bacterial infections. Of greatest concern are infections caused by extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-EC). The aim of our study was to evaluate the prevalence and molecular characteristics of ESBL-EC isolated over an 18-year pre-COVID period from lower respiratory tract (LRT) samples collected from selected Slovenian hospitals. OBJECTIVES AND METHODS: All isolates were identified by MALDI-TOF and phenotypically confirmed as ESBLs by a disk diffusion assay. Using a PCR approach, 487 non-repetitive isolates were assigned to phylogroups, sequence type groups, and clonal groups. Isolates were also screened for virulence-associated genes (VAGs) and antimicrobial resistance genes. RESULTS: The prevalence of ESBL-EC isolates from LRT in a large university hospital was low (1.4%) in 2005 and increased to 10.8% by 2019. The resistance profile of 487 non-repetitive isolates included in the study showed a high frequency of group 1 blaCTX-M (77.4%; n = 377), blaTEM (54.4%; n = 265) and aac(6')-Ib-cr (52%; n = 253) genes and a low proportion of blaSHV and qnr genes. Isolates were predominantly assigned to phylogroup B2 (73.1%; n = 356), which was significantly associated with clonal group ST131. The ST131 group accounted for 67.6% (n = 329) of all isolates and had a higher number of virulence factor genes than the non-ST131 group. The virulence gene profile of ST131 was consistent with that of other extraintestinal pathogenic E. coli (ExPEC) strains and was significantly associated with ten of sixteen virulence factor genes tested. Using ERIC-PCR fingerprinting, isolates with the same ERIC-profile in samples from different patients, and at different locations and sampling dates were confirmed, indicating the presence of "hospital-adapted" strains. CONCLUSION: Our results suggest that the ESBL-EC isolates from LRT do not represent a specific pathotype, but rather resemble other ExPEC isolates, and may be adapted to the hospital environment. To our knowledge, this is the first study of ESBL-EC isolated from LRT samples collected over a long period of time.

The evolution of cavitation in narrow soft-solid wedge geometry mimicking periodontal and peri-implant pockets.

In periodontology and implantology, laser-induced cavitation has not yet been used to treat biofilm-related problems. In this study we have checked how soft tissue affects the evolution of cavitation in a wedge model representing periodontal and peri-implant pocket geometry. One side of the wedge model was composed of PDMS mimicking soft periodontal or peri-implant biological tissue, the other side was composed of glass mimicking hard tooth root or implant surface, which allowed observations of the cavitation dynamics with an ultrafast camera. Different laser pulse modalities, PDMS stiffness, and irrigants were tested for their effect on the evolution of cavitation in the narrow wedge geometry. The PDMS stiffness varied in a range that corresponds to severely inflamed, moderately inflamed, or healthy gingival tissue as determined by a panel of dentists. The results imply that deformation of the soft boundary has a major effect on the Er:YAG laser-induced cavitation. The softer the boundary, the less effective the cavitation. We show that in a stiffer gingival tissues model, photoacoustic energy can be guided and focused at the tip of the wedge model, where it enables generation of secondary cavitation and more effective microstreaming. The secondary cavitation was absent in severely inflamed gingival model tissue, but could be induced with a dual-pulse AutoSWEEPS laser modality. This should in principle increase cleaning efficiency in the narrow geometries such as those found in the periodontal and peri-implant pockets and may lead to more predictable treatment outcomes.

Small Prokaryotic DNA-Binding Proteins Protect Genome Integrity throughout the Life Cycle.

Genomes of all organisms are persistently threatened by endogenous and exogenous assaults. Bacterial mechanisms of genome maintenance must provide protection throughout the physiologically distinct phases of the life cycle. Spore-forming bacteria must also maintain genome integrity within the dormant endospore. The nucleoid-associated proteins (NAPs) influence nucleoid organization and may alter DNA topology to protect DNA or to alter gene expression patterns. NAPs are characteristically multifunctional; nevertheless, Dps, HU and CbpA are most strongly associated with DNA protection. Archaea display great variety in genome organization and many inhabit extreme environments. As of yet, only MC1, an archaeal NAP, has been shown to protect DNA against thermal denaturation and radiolysis. ssDNA are intermediates in vital cellular processes, such as DNA replication and recombination. Single-stranded binding proteins (SSBs) prevent the formation of secondary structures but also protect the hypersensitive ssDNA against chemical and nuclease degradation. Ionizing radiation upregulates SSBs in the extremophile Deinococcus radiodurans.

High virucidal potential of novel ceramic-metal composites fabricated via hybrid selective laser melting and spark plasma sintering routes.

In this work, we combine selective laser melting (SLM) and spark plasma sintering (SPS) to fabricate new materials with high virucidal potential. Various bioactive disc-shaped ceramics, metal alloys, and composites were fabricated and tested against bacteriophage Phi6-a model system for RNA-enveloped viruses. We prepared silver-doped titanium dioxide (TiO2 + 2.5‒10% Ag), copper-doped titanium dioxide (TiO2 + 2.5‒10% Cu), Cu2NiSiCr, and Cu15Ni8Sn composite materials (metal lattices filled with ceramics). The virucidal tests of the ceramic and metal powders were performed in buffered suspensions, while the surfaces of the discs were tested by swabbing. The results show that the virus titer on the TiO2 + 10% Ag ceramic and CuNi2SiCr metal discs decreased by 4 logs after 15 min of exposure to the surfaces compared to the control ceramic and steel discs. We show that SLM 3D printed pre-alloyed CuNi2SiCr filled with bioactive TiO2 + 10% Ag nanopowders and sintered by the SPS process combines the simplicity of printing with the strength and virucidal properties of Ag and Cu materials. The proposed new virucidal materials were also used for the fabrication of prototype elevator buttons.

Phi 6 Bacteriophage Inactivation by Metal Salts, Metal Powders, and Metal Surfaces.

The interaction of phages with abiotic environmental surfaces is usually an understudied field of phage ecology. In this study, we investigated the virucidal potential of different metal salts, metal and ceramic powders doped with Ag and Cu ions, and newly fabricated ceramic and metal surfaces against Phi6 bacteriophage. The new materials were fabricated by spark plasma sintering (SPS) and/or selective laser melting (SLM) techniques and had different surface free energies and infiltration features. We show that inactivation of Phi6 in solutions with Ag and Cu ions can be as effective as inactivation by pH, temperature, or UV. Adding powder to Ag and Cu ion solutions decreased their virucidal effect. The newly fabricated ceramic and metal surfaces showed very good virucidal activity. In particular, 45%TiO2 + 5%Ag + 45%ZrO2 + 5%Cu, in addition to virus adhesion, showed virucidal and infiltration properties. The results indicate that more than 99.99% of viruses deposited on the new ceramic surface were inactivated or irreversibly attached to it.

Association between pre-pregnancy body weight and dietary pattern with large-for-gestational-age infants in gestational diabetes.

BACKGROUND: Both obesity and gestational diabetes (GDM) are associated with adverse outcomes. Diet during pregnancy impacts weight gain and fetal growth. Therefore, we aimed to explore non-pharmacological treatment success depending on pre-pregnancy body weight and its association with large for gestational age (LGA) infants in women with GDM. METHODS: In our observational study we investigated 57 singleton pregnant women with GDM. All women received standard treatment, including healthy diet education and regular medical checkups. Data were collected through blood analysis, medical records and questionnaires assessing diet before conception and during pregnancy. Differences in dietary patterns were compared in normal weight and overweight/obese group using Mann-Whitney U, Wilcoxon Signed Rank Test or Kruskal-Wallis test, as appropriate. Logistic regression was used for prediction of LGA. p-value less than 0.05 was used for statistical significance. RESULTS: Preconceptionally, the Mann-Whitney U test showed that the normal-weight group (n = 41) more frequently consumed fruits (U = 116.5, p < 0.001), eggs (U = 189.5, p = 0.02), cheese (U = 148.0, p = 0.003) compared to the overweight/obese group (n = 16), that consumed more beef (U = 407.0, p = 0.03) and low-calorie beverages (U = 397.0, p = 0.05). During pregnancy both groups improved their diet, with no differences detected. Personality types differed only preconceptionally with regard to healthy diet. Excessive gestational weight gain did not significantly differ between body-weight groups (16.6% vs. 23.1%), neither did the incidence of LGA infants (46.2% vs. 43.8%). Significant predictors of LGA were paternal height (OR = 1.12, 95% CI 1.01-1.23), 3rd trimester HbA1c (OR = 0.50, 95% CI 0.26-0.97), unemployment (OR = 4.80, 95% CI 1.12-20.61) and diet improvement during pregnancy (OR = 1.19, 95% CI 1.02-1.39). After adjustment improvement in diet was no longer a significant predictor for LGA. CONCLUSION: Even though dietary patterns of the participants significantly improved during pregnancy, LGA infants were born independently of pre-pregnancy weight or diet and despite good glycemic control. Further research is needed to explore social determinants of health and whether solutions outside the health sector could provide efficient means in preventing adverse pregnancy outcomes as well as improving metabolic health.

The Escherichia coli colibactin resistance protein ClbS is a novel DNA binding protein that protects DNA from nucleolytic degradation.

Cells employ specific and nonspecific mechanisms to protect their genome integrity against exogenous and endogenous factors. The clbS gene is part of the polyketide synthase machinery (pks genomic island) encoding colibactin, a genotoxin implicated in promoting colorectal cancer. The pks is found among the Enterobacteriaceae, in particular Escherichia coli strains of the B2 phylogenetic group. Several resistance mechanisms protect toxin producers against toxicity of their products. ClbS, a cyclopropane hydrolase, was shown to confer colibactin resistance by opening its electrophilic cyclopropane ring. Here we report that ClbS sustained viability and enabled growth also of E. coli expressing another genotoxin, the Usp nuclease. The recA::gfp reporter system showed that ClbS protects against Usp induced DNA damage. To elucidate the mechanism of ClbS mediated protection, we studied the DNA binding ability of the ClbS protein. We show that ClbS directly interacts with single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), whereas ssDNA seems to be the preferred substrate. Thus, the ClbS DNA-binding characteristics may serve bacteria to protect their genomes against DNA degradation.