Influence of bioceramic sealer residues on the adhesion interface with a universal adhesive in the etch-and-rinse strategy, based on the application protocol and evaluation time.

The aim of this study was to evaluate the effects of the application of universal adhesive in the etch-and-rinse (ER) strategy with a manual brush (MB) or rotary brush (RB) in adhesion to dentin impregnated (WB) or not (NB) with bioceramic sealer, at 24 h and 1 year. Eighty-eight crowns of bovine incisors were divided into four groups (n=22): WB-MB, WB-RB, NB-MB, NB-RB. After the restorative protocol, dentin penetrability was evaluated by confocal microscopy. Bond strength was evaluated by microtensile test. Data were analyzed using one-way ANOVA/Tukey tests (α=0.05). NB-RB and WB-RB exhibited greater extensions of resin tags in dentin. At 24 h, WB-RB and NB-MB showed the highest and lowest values of bond strength, respectively. At 1 year, WB-MB and WB-RB demonstrated the highest bond strength values. The RB increases the formation of resin tags and residues of bioceramic sealer provides higher bond strength in the ER strategy.

The detection of KPC-2, NDM-1, and VIM-2 carbapenemases in international clones isolated from fresh vegetables highlights an emerging food safety issue.

Resistance to carbapenems emerged in clinical settings and has rapidly spread to other sectors, such as food and the environment, representing a One Health problem. In this regard, vegetables contaminated by critical priority pathogens have raised global concerns. Here, we have performed a whole-genome sequence-based analysis of extensively drug-resistant Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa strains isolated from cabbage, spinach, and lettuce, respectively. Genomic analysis revealed the emergence of international and high-risk clones belonging to ST340, ST155, and ST233, harboring a broad resistome to clinically important antimicrobials. In this context, K. pneumoniae, E. coli, and P. aeruginosa strains carried blaKPC-2, blaNDM-1, and blaVIM-2, respectively. The blaKPC-2 gene with a non-Tn4401 element (NTEKPC-Ic) was located on an IncX3-IncU plasmid, while the blaVIM-2 gene was associated with a Tn402-like class 1 integron, In559, on the chromosome. Curiously, the blaNDM-1 gene coexisted with the blaPER-2 gene on an IncC plasmid and the regions harboring both genes contained sequences of Tn3-like element ISKox2-like family transposase. Comparative genomic analysis showed interspecies and clonal transmission of carbapenemase-encoding genes at the human-animal-environmental interface. These findings raise a food safety alert about hospital-associated carbapenemase producers, supporting that fresh vegetables can act as a vehicle for the spread of high-risk clones.

Evaluation of bond strength in customizing fiber post using translucent resins photoactivated by different light-emitting diodes.

Aims: The objective of the study was to evaluate two translucent resin composite systems for customizing light-polymerized fiber posts with light-emitting diode (LED) curing units regarding adhesion using conventional cement at 24 h and 6 months. Settings and Design: This was an experimental in vitro study. Methods: Forty roots were prepared and divided into four groups (n = 10): ZV-Z350 resin and LED Valo; ZR-Z350 resin and LED Radii-Cal; OV-Opallis resin and LED Valo; and OR-Opallis resin and LED Radii-Cal. The fiber post was customized and cemented with conventional resin cement and was photoactivated by two different sources. Statistical Analysis Used: The data were subjected to two-way ANOVA and Tukey's post hoc tests (P = 0.05). The data regarding the pattern of adhesive failures were described in terms of the frequency of occurrence in each third of the postspace. Results: Both at 24 h and 6 months, bond strength among groups was similar, regardless of the analyzed postspace 3rd (P > 0.05). Adhesive failure Type 4 was the most frequent in all thirds. Conclusions: Customizing the fiber post with Z350 and Opallis has the same effect on bond strength and adhesive failure pattern, regardless of the LED curing units used for photopolymerization.

Emergence of carbapenem-resistant Klebsiella pneumoniae species complex from agrifood systems: detection of ST6326 co-producing KPC-2 and NDM-1.

BACKGROUND: Klebsiella pneumoniae species complex (KpSC) is an important disseminator of carbapenemase-encoding genes, mainly blaKPC-2 and blaNDM-1, from hospitals to the environment. Consequently, carbapenem-resistant strains can be spread through the agrifood system, raising concerns about food safety. This study therefore aimed to isolate carbapenem-resistant KpSC strains from the agricultural and environmental sectors and characterize them using phenotypic, molecular, and genomic analyses. RESULTS: Klebsiella pneumoniae and Klebsiella quasipneumoniae strains isolated from soils used for lemon, guava, and fig cultivation, and from surface waters, displayed an extensive drug-resistance profile and carried blaKPC-2, blaNDM-1, or both. In addition to carbapenemase-encoding genes, KpSC strains harbor a broad resistome (antimicrobial resistance and metal tolerance) and present putative hypervirulence. Soil-derived K. pneumoniae strains were assigned as high-risk clones (ST11 and ST307) and harbored the blaKPC-2 gene associated with Tn4401b and Tn3-like elements on IncN-pST15 and IncX5 plasmids. In surface waters, the coexistence of blaKPC-2 and blaNDM-1 genes was identified in K. pneumoniae ST6326, a new carbapenem-resistant regional Brazilian clone. In this case, blaKPC-2 with Tn4401a isoform and blaNDM-1 associated with a Tn125-like transposon were located on different plasmids. Klebsiella quasipneumoniae ST526 also presented the blaNDM-1 gene associated with a Tn3000 transposon on an IncX3 plasmid. CONCLUSION: These findings provide a warning regarding the transmission of carbapenemase-positive KpSC across the agricultural and environmental sectors, raising critical food safety and environmental issues. © 2024 Society of Chemical Industry.

Genetic diversity of KPC-2-producing Klebsiella pneumoniae complex from aquatic ecosystems.

During the COVID-19 pandemic, the occurrence of carbapenem-resistant Klebsiella pneumoniae increased in human clinical settings worldwide. Impacted by this increase, international high-risk clones harboring carbapenemase-encoding genes have been circulating in different sources, including the environment. The blaKPC gene is the most commonly disseminated carbapenemase-encoding gene worldwide, whose transmission is carried out by different mobile genetic elements. In this study, blaKPC-2-positive Klebsiella pneumoniae complex strains were isolated from different anthropogenically affected aquatic ecosystems and characterized using phenotypic, molecular, and genomic methods. K. pneumoniae complex strains exhibited multidrug-resistant and extensively drug-resistant profiles, spotlighting the resistance to carbapenems, ceftazidime-avibactam, colistin, and tigecycline, which are recognized as last-line antimicrobial treatment options. Molecular analysis showed the presence of several antimicrobial resistance, virulence, and metal tolerance genes. In-depth analysis showed that the blaKPC-2 gene was associated with three different Tn4401 isoforms (i.e., Tn4401a, Tn4401b, and Tn4401i) and NTEKPC elements. Different plasmid replicons were detected and a conjugative IncN-pST15 plasmid harboring the blaKPC-2 gene associated with Tn4401i was highlighted. K. pneumoniae complex strains belonging to international high-risk (e.g., ST11 and ST340) and unusual clones (e.g., ST323, ST526, and ST4216) previously linked to clinical settings. In this context, some clones were reported for the first time in the environmental sector. Therefore, these findings evidence the occurrence of carbapenemase-producing K. pneumoniae complex strains in aquatic ecosystems and contribute to the monitoring of carbapenem resistance worldwide.

A skeleton in a huff: insights in etiologies of osteosclerosis.

A 30-yr-old man developed right lower leg pain and a palpable solid mass. Radiographic imaging revealed a periosteal reaction with an exostotic mass arising from the right distal fibula. Generalized skeletal osteosclerosis with periosteal reaction was discovered on a radiographic skeletal survey. A biopsy of the right fibular mass revealed reactive woven bone. The patient was referred to a metabolic bone disease clinic, where laboratory values were consistent with secondary hyperparathyroidism and increased bone turnover. A DXA bone density scan revealed high bone density, with an L1-4 spine Z-score of +9.3, a left femoral neck Z-score of +8.5, and a total hip Z-score of +6.5. A dental exam revealed generalized gingival inflammation, teeth mobility, generalized horizontal alveolar bone loss and widening of the periodontal ligament space, increased bone density around the teeth, and thickening of the radicular lamina dura. An extensive evaluation was performed, with the result of a single test revealing the diagnosis. The differential diagnoses of osteosclerosis affecting the skeleton, teeth, and oral cavity are discussed.

A 30-yr-old man developed, over a short period, pain in his lower right leg accompanied by a hard mass. He also reported weight loss and night sweats for the past 6 months. After evaluation by his primary physician, an X-ray was ordered that reported a bony mass arising from the right fibula bone. A biopsy was performed of the mass, but no evidence of cancer or any other specific abnormality was found. The patient was then referred to a bone disease specialty clinic. Laboratory tests revealed a large increase in how quickly the patient's skeleton was remodeling, affecting the balance of bone formation and removal involved in maintaining a healthy skeleton. A bone density scan reported that the patient had very dense bones. Other unusual changes were also discovered in a dental exam, suggesting bone thickening. After an extensive evaluation, a single blood test revealed the cause of the fibular bone mass and dense bones.

Evaluation of TRIM63 RNA in situ hybridization (RNA-ISH) as a potential biomarker for alveolar soft-part sarcoma (ASPS).

Alveolar soft-part sarcoma (ASPS) is a rare soft tissue tumor with a broad morphologic differential diagnosis. While histology and immunohistochemistry can be suggestive, diagnosis often requires exclusion of other entities followed by confirmatory molecular analysis for its characteristic ASPSCR1-TFE3 fusion. Current stain-based biomarkers (such as immunohistochemistry for cathepsin K and TFE3) show relatively high sensitivity but may lack specificity, often showing staining in multiple other entities under diagnostic consideration. Given the discovery of RNA in situ hybridization (RNA-ISH) for TRIM63 as a sensitive and specific marker of MiTF-family aberration renal cell carcinomas, we sought to evaluate its utility in the workup of ASPS. TRIM63 RNA-ISH demonstrated high levels (H-score greater than 200) of expression in 19/20 (95%) cases of ASPS (average H-score 330) and was weak or negative in cases of paraganglioma, clear cell sarcoma, rhabdomyosarcoma, malignant epithelioid hemangioendothelioma, as well as hepatocellular and adrenal cortical carcinomas. Staining was also identified in tumors with known subsets characterized by TFE3 alterations such as perivascular epithelioid cell neoplasm (PEComa, average H-score 228), while tumors known to exhibit overexpression of TFE3 protein without cytogenetic alterations, such as melanoma and granular cell tumor, generally showed less TRIM63 ISH staining (average H-scores 147 and 96, respectively). Quantitative assessment of TRIM63 staining by RNA-ISH is potentially a helpful biomarker for tumors with molecular TFE3 alterations such as ASPS.

Formulations of NaOCl-based in liquid, gel form or with surfactants on dentin deproteinization before fiber post cementation.

The objective of the study was to evaluate the effects of dentin deproteinization protocols for post space using different formulations containing sodium hypochlorite before fiber post cementation with self-adhesive resinous cement. The groups were divided according to the irrigation protocol (DWC, SHS, SHT and SHG). The residue cleanliness, bond strength, adhesive failure pattern, and tag formation at the adhesive interface between the self-adhesive cement and the dentin were evaluated. For this, analysis in scanning electron microscope, push-out test and confocal laser scanning microscopy were performed. The SHT protocol showed the highest residue cleanliness on the dentin surface of the post space (p<0.05). In addition, SHT protocol showed highest bond strength and tag formation in the cervical and middle thirds (p<0.05). Dentin deproteinization with sodium hypochlorite with surfactant provided the best dentin cleaning of residues, bond strength and tag formation after cementation of the fiber post with self-adhesive cement.

Influence of using different translucent composite resins for customizing fiber post on the bond strength of self-adhesive cement to root dentin.

This study evaluated the influence of different translucent resins (Z350 and Opallis) for customizing fiber posts and light-curing the cementation system using different LED equipment (V, Valo or R, Radii-Cal) on the bond strength and adhesive failure pattern at 24 h and 6 months. Eighty roots were prepared and divided into 4 groups (n=20): ZV (Z350 resin and LED Valo), ZR (Z350 resin and LED Radii-Cal), OV (Opallis resin and LED Valo), OR (Opallis resin and LED Radii-Cal). After post space preparation, the fiber post was customized and cemented with self-adhesive cement and light-curing using V or R LED equipment. Bond strength values were submitted to 2-way ANOVA test. ZV and ZR showed higher bond strength values than the other groups at 6 months of evaluation (p<0.05). The Z350 resin has a favorable influence on the bond strength of self-adhesive cement to root dentin, regardless of the LED polymerization equipment used.

Vaccinating international seafarers during the COVID-19 pandemic.

Despite their key role in the distribution of essential goods, seafarers were often overlooked in international COVID-19 responses. Border closures isolated them at sea, often for months beyond contract. Distributing vaccines to seafarers was inconsistent and often depended on the creativity of charities and industry and labour stakeholders. Even seafarers who received vaccinations often had them unrecognised in the ports they visited. These oversights should not be repeated in preparation for future pandemics. As a vulnerable population and as key international workers, seafarers' health should be prioritised by national and international health policy.

Effect of sodium hypochlorite-based formulations on the adhesion interface after fiber post cementation.

The study evaluated the persistence of residues, bond strength and tags formation of a cementation system after post-space irrigation with different sodium hypochlorite-based irrigants. The groups were divided according to the irrigation protocol (DW: distilled water; SHS: sodium hypochlorite solution; SHG: sodium hypochlorite gel; and, SHT: sodium hypochlorite with surfactant). Forty roots (n=10) were used to evaluate the persistence of residues by scanning electron microscopy. Other forty roots were used to the push-out bond strength test, failure mode and tag formation analyses. Non-parametric data were submitted to Kruskal Wallis and Dunn tests, while parametric data were evaluated by one-way ANOVA. No difference was observed in the persistence of residues. DW showed the highest bond strength and tag formation. Type 2 failure mode was the most frequent in the experimental groups. Compared to control, SHT showed the best performance, since less negative effects on adhesive interface was observed.

Genomic features of an extensively drug-resistant and NDM-1-positive Klebsiella pneumoniae ST340 isolated from river water.

The environmental contamination plays a significant role in the emergence of antimicrobial resistance. In this study, we report a genomic analysis of an extensively drug-resistant and blaNDM-1-producing Klebsiella pneumoniae (EW807) strain recovered from a surface water sample. Strain EW807 belonged to sequence type (ST) 340 and serotype O4:KL15, a high-risk clone of the clonal group 258. This strain carried a broad resistome, including blaNDM-1 and blaCTX-M-15. The core genome multilocus sequence typing phylogenetic analysis revealed that the EW807 strain was most related to strains from Brazil and the USA. An IncX3 plasmid was identified harboring the blaNDM-1 gene, while an IncFIB(K) plasmid was detected carrying the blaCTX-M-15 in addition to multidrug resistance and multimetal tolerance regions. IncX3 and IncFIB(K) plasmids shared high similarity with plasmids from a human in China and a dog in Brazil, respectively. The regions harboring the blaNDM-1 and blaCTX-M-15 genes contained sequences from the Tn3 family. These findings suggest that IncX3 plasmid could play a role in the spread of NDM-1 in a post-pandemic scenario. To the best of our knowledge, this is the first report of blaNDM-1-producing K. pneumoniae ST340 O4:KL15 strain in the environment. Therefore, the presence of high-risk clones of K. pneumoniae carrying carbapenemases in the environment requires strict surveillance.

Klebsiella quasipneumoniae subsp. similipneumoniae ST1859 O5:KL35 from Soil: First Report of qnrE1 in the Environment.

A Klebsiella quasipneumoniae subsp. similipneumoniae strain, named S915, belonging to the ST1859 O5:KL35, and harboring the plasmid-mediated quinolone resistance qnrE1 gene, was isolated from a soil sample cultivated with lettuce in Brazil. The core genome multilocus sequence typing analysis revealed that S915 strain was most related to a clinical strain of Brazil. Comparative genomic analysis showed that ST1859 O5:KL35 strains have been circulating in clinical settings and are closely related to multidrug resistance and multimetal tolerance. Strain S915 presented a plasmid contig co-harboring the qnrE1 gene and tellurite tolerance operon. The region harboring the qnrE1 gene (ISEcp1-qnrE1-araJ-ahp) shared high similarity with others from infected humans, ready-to-eat dish, and food-producing animals in Brazil. This is the first report of the plasmid-mediated qnrE1 gene in the environment. Our findings evidence the initial dissemination of the qnrE1 gene in the environment by the introduction of a clinical strain, which may be spread to different sectors, representing a One Health challenge.

COVID-19 on board a cruise ship: medical management.

BACKGROUND: First, this analysis was conducted to study a coronavirus disease 2019 (COVID-19) cluster dynamic on a cruise ship in order to allow the ship physician to anticipate the duration and importance of the contaminations. Secondly, the author tries to find out if the closed environment on board allows specific conclusions about epidemic dynamics and preventative measures. MATERIALS AND METHODS: From a personal epidemiological compendium done by himself on board the author analysed different epidemic curves identified on board other ships and compared them to the epidemiologic data from the different COVID-19 contamination waves in France since 2020. All crew members were submitted to polymerase chain reaction tests on D2, D5, D8 and D15 and symptomatic cases were tested on on-board devices in the meantime. An excel file called "Log Covid" allowed for daily reporting to the ship-owner on the epidemic dynamics and the prospects on the end of crises in order to anticipate the resumption of the business in the best conditions. The jobs on board, age and geographic origin of the contaminated people were analysed, as well as their vaccination status. RESULTS: Out of a total of 118 crew members, 61 (52%) sailors were contaminated in 8 days. The symptoms were benign (pharyngitis, headaches, feverish state); no serious form of illness were reported. The passengers were repatriated to France at the earliest stage. The epidemic phase occurred in a 15-day window. The first 8 days corresponded to the ascending phase, then a faster phase of epidemic decrease of 7 days. Similarities emerged between the epidemic dynamics of this virus and other contaminations on cruise ships and epidemic phases on land in spite of important differences in numbers. CONCLUSIONS: This study can allow a ship's doctor to better understand the viral dynamics in case of a COVID- 19 cluster and to anticipate the exit of the crisis. Repeated tests during the active phase of the epidemic are necessary in case of a large cluster to know where to place oneself on a typical epidemic curve. Isolation and barrier measures advised by the ship's doctor remain the only weapons that can limit its magnitude.

Surface and intra-pulpal temperature variation during tooth whitening photoactivated with LED/laser.

This study evaluated the variation of surface and intra-pulpal temperature, during bleaching protocol, using LED/laser. The 35% (HP35), 15% (HP15) and 6% (HP6) gels were used associated with LED/laser applied every 1 min for 30 min in a human canine. The evaluation of surface temperature variation (∆Ts) was performed using a pHmeter and the intra-pulpal temperature variation (∆Ti) was performed using a digital thermometer, at times of 1-, 5-, 10- 15- and 30-min. Statistical analysis was performed using the two-way repeated measures ANOVA test and Bonferroni post-hoc test was used at a significance level of 5%. HP35 and HP15 showed greater temperature variation than HP6 up to 10 min of surface evaluation, showing no differences between them. In the intra-pulpal evaluation, no group showed differences throughout the procedure.

Patterns of antimicrobial resistance and metal tolerance in environmental Pseudomonas aeruginosa isolates and the genomic characterization of the rare O6/ST900 clone.

Pseudomonas aeruginosa can harbor several virulence and antimicrobial resistance genes (ARGs). In this regard, virulent and multidrug-resistant (MDR) P. aeruginosa strains are closely related to severe infections. In addition, this species can also carry metal tolerance genes, selecting mainly antimicrobial-resistant strains. The action of several pollutants on the environment may favor the occurrence of antimicrobial-resistant and metal-tolerant strains. Therefore, the aim of this study was to characterize potentially pathogenic, antimicrobial-resistant, and/or metal-tolerant P. aeruginosa isolates from different environmental samples (waters, soils, sediments, or sands) and to perform a whole-genome sequence-based analysis of a rare clone from residual water. Environmental isolates carried virulence genes related to adherence, invasion, and toxin production, and 79% of the isolates harbored at least five virulence genes. In addition, the isolates were resistant to different antimicrobials, including important antipseudomonal agents, and 51% of them were classified as MDR, but only ARGs associated with aminoglycoside resistance were found. Furthermore, some isolates were tolerant mainly to copper, cadmium, and zinc, and presented metal tolerance genes related to these compounds. Whole-genome characterization of an isolate with unique phenotype with simultaneous resistance to antimicrobials and metals showed nonsynonymous mutations in different antimicrobial resistance determinants and revealed a classification of O6/ST900 clone as rare, potentially pathogenic, and predisposed to acquire multidrug resistance genes. Therefore, these results draw attention to the dissemination of potentially pathogenic, antimicrobial-resistant, and metal-tolerant P. aeruginosa isolates in environmental niches, alerting to a potential risk mainly to human health.

Multidrug-resistant Shiga toxin-producing Escherichia coli and hybrid pathogenic strains of bovine origin.

Antimicrobial-resistant Escherichia coli strains have been circulating in various sectors and can be cross-transferred between them. Among pathogenic E. coli strains, Shiga toxin-producing E. coli (STEC) and hybrid pathogenic E. coli (HyPEC) emerged as responsible for outbreaks worldwide. As bovine are reservoir of STEC strains, these pathogens primarily spread to food products, exposing humans to risk. Therefore, this study aimed to characterize antimicrobial-resistant and potentially pathogenic E. coli strains from fecal samples of dairy cattle. In this regard, most E. coli strains (phylogenetic groups A, B1, B2, and E) were resistant to ß-lactams and non-ß-lactams and were classified as multidrug-resistant (MDR). Antimicrobial resistance genes (ARGs) related to multidrug resistance profiles were detected. Furthermore, mutations in fluoroquinolone and colistin resistance determinants were also identified, highlighting the deleterious mutation His152Gln in PmrB that may have contributed to the high level (> 64 mg/L) of colistin resistance. Virulence genes of diarrheagenic and extraintestinal pathogenic E. coli (ExPEC) pathotypes were shared among strains and even within the same strain, evidencing the presence of HyPEC (i.e., ExPEC/STEC), which were assigned as unusual B2-ST126-H3 and B1-ST3695-H31. These findings provide phenotypic and molecular data of MDR, ARGs-producing, and potentially pathogenic E. coli strains in dairy cattle, contributing to the monitoring of antimicrobial resistance and pathogens in healthy animals and alerting to potential bovine-associated zoonotic infections.