Evaluation of coccidia DNA in irrigation pond water and wastewater sludge associated with Cyclospora cayetanensis 18S rRNA gene qPCR detections.

The coccidian parasite Cyclospora cayetanensis is the causative agent for foodborne outbreaks of cyclosporiasis disease and multiple annual fresh produce recalls. The aim of this study was to identify potential cross-reacting species for the C. cayetanensis 18S rRNA and MIT1C gene target real-time quantitative polymerase chain reaction (qPCR) assays. The environmental samples evaluated were irrigation pond water, produce wash water, and wastewater treatment sludge from a previous study with qPCR detections of C. cayetanensis by the 18S rRNA gene target qPCR. From these samples, longer regions of the 18S rRNA gene and the mitochondrial cytochrome c oxidase subunit III gene (cox3) were sequenced. Of 65 irrigation pond water samples with positive test results using the C. cayetanensis 18S rRNA gene qPCR assay, none had MIT1C qPCR assay detections or sequences that clustered with C. cayetanensis based on sequencing of the cox3 and 18S rRNA gene. Sequences from these samples clustered around coccidia sequences found in bird, fish, reptile, and amphibian hosts. Of 26 sludge samples showing detections by either qPCR assay, 14 (54%) could be confirmed as containing C. cayetanensis by sequencing of cox3 and 18S rRNA gene regions. In three of the remaining sludge samples, sequenced reads clustered with coccidia from rodents. This study demonstrated that caution should be taken when interpreting qPCR C. cayetanensis detection data in environmental samples and sequencing steps will likely be needed for confirmation. IMPORTANCE: Fresh produce is a leading transmission source in cyclosporiasis outbreaks. It is therefore essential to understand the role that produce-growing environments play in the spread of this disease. To accomplish this, sensitive and specific tests for environmental and irrigation waters must be developed. Potential cross-reactions of Cyclospora cayetanensis real-time quantitative polymerase chain reaction (qPCR) assays have been identified, hindering the ability to accurately identify this parasite in the environment. Amplicon sequencing of the cox3 and 18S rRNA genes revealed that all irrigation pond water and two sludge samples that initially detected C. cayetanensis by qPCR were most likely cross-reactions with related coccidian organisms shed from birds, fish, reptiles, amphibians, and rodents. These results support that a single testing method for environmental samples is likely not adequate for sensitive and specific detection of C. cayetanensis.

Prevalence of STEC virulence markers and Salmonella as a function of abiotic factors in agricultural water in the southeastern United States.

Fresh produce can be contaminated by enteric pathogens throughout crop production, including through contact with contaminated agricultural water. The most common outbreaks and recalls in fresh produce are due to contamination by Salmonella enterica and Shiga toxin-producing E. coli (STEC). Thus, the objectives of this study were to investigate the prevalence of markers for STEC (wzy, hly, fliC, eaeA, rfbE, stx-I, stx-II) and Salmonella (invA) in surface water sources (n = 8) from produce farms in Southwest Georgia and to determine correlations among the prevalence of virulence markers for STEC, water nutrient profile, and environmental factors. Water samples (500 mL) from eight irrigation ponds were collected from February to December 2021 (n = 88). Polymerase chain reaction (PCR) was used to screen for Salmonella and STEC genes, and Salmonella samples were confirmed by culture-based methods. Positive samples for Salmonella were further serotyped. Particularly, Salmonella was detected in 6/88 (6.81%) water samples from all ponds, and the following 4 serotypes were detected: Saintpaul 3/6 (50%), Montevideo 1/6 (16.66%), Mississippi 1/6 (16.66%), and Bareilly 1/6 (16.66%). Salmonella isolates were only found in the summer months (May-Aug.). The most prevalent STEC genes were hly 77/88 (87.50%) and stx-I 75/88 (85.22%), followed by fliC 54/88 (61.63%), stx-II 41/88 (46.59%), rfbE 31/88 (35.22%), and eaeA 28/88 (31.81%). The wzy gene was not detected in any of the samples. Based on a logistic regression analysis, the odds of codetection for STEC virulence markers (stx-I, stx-II, and eaeA) were negatively correlated with calcium and relative humidity (p < 0.05). A conditional forest analysis was performed to assess predictive performance (AUC = 0.921), and the top predictors included humidity, nitrate, calcium, and solar radiation. Overall, information from this research adds to a growing body of knowledge regarding the risk that surface water sources pose to produce grown in subtropical environmental conditions and emphasizes the importance of understanding the use of abiotic factors as a holistic approach to understanding the microbial quality of water.

Sources and Prevalence of Cyclospora cayetanensis in Southeastern U.S. Growing Environments.

Recent cyclosporiasis outbreaks associated with fresh produce grown in the United States highlight the need to better understand Cyclospora cayetanensis prevalence in U.S. agricultural environments. In this study, C. cayetanensis occurrence was assessed in municipal wastewater sludge, on-farm portable toilets, irrigation pond water, and spent packing house dump tank water in a Southeastern Georgia growing region over two years. Detection of the C. cayetanensis 18S rRNA qPCR gene target in pond samples was 0%, 28%, and 42% (N = 217) depending on the detection definition used, and ≤1% in dump tank samples (N = 46). However, no qPCR detections were confirmed by sequencing, suggesting false detection occurred due to cross-reactions. C. cayetanensis qPCR detections were confirmed in 9% of wastewater sludge samples (N = 76). The human-specific fecal markers HF183 and crAssphage were detected in 33% and 6% of pond samples, respectively, and 4% and 0% of dump tank samples, respectively. Despite community Cyclospora shedding and evidence of human fecal contamination in irrigation water, there was no correlation between C. cayetanensis and HF183 qPCR detections, further supporting that 18S gene target qPCR amplifications were due to cross-reactions. When evaluating C. cayetanensis qPCR environmental detection data, the impact of assay specificity and detection criteria should be considered. Moreover, additional sequence-based testing may be needed to appropriately interpret Cyclospora qPCR environmental data.

Convergent Mutations and Single Nucleotide Variants in Mitochondrial Genomes of Modern Humans and Neanderthals.

The genetic contributions of Neanderthals to the modern human genome have been evidenced by the comparison of present-day human genomes with paleogenomes. Neanderthal signatures in extant human genomes are attributed to intercrosses between Neanderthals and archaic anatomically modern humans (AMHs). Although Neanderthal signatures are well documented in the nuclear genome, it has been proposed that there is no contribution of Neanderthal mitochondrial DNA to contemporary human genomes. Here we show that modern human mitochondrial genomes contain 66 potential Neanderthal signatures, or Neanderthal single nucleotide variants (N-SNVs), of which 36 lie in coding regions and 7 result in nonsynonymous changes. Seven N-SNVs are associated with traits such as cycling vomiting syndrome, Alzheimer's disease and Parkinson's disease, and two N-SNVs are associated with intelligence quotient. Based on recombination tests, principal component analysis (PCA) and the complete absence of these N-SNVs in 41 archaic AMH mitogenomes, we conclude that convergent evolution, and not recombination, explains the presence of N-SNVs in present-day human mitogenomes.

Yellow fever neutralizing antibody seroprevalence proportion and titers in previously vaccinated adults with chronic kidney disease.

Studies on yellow fever vaccine (YF) in chronic kidney disease (CKD) patients are scarce. This cross-sectional study aimed to evaluate YF neutralizing antibody seroprevalence and titers in previously vaccinated adults with CKD, on dialysis (D-CKD) or not (ND-CKD), compared to healthy persons. The micro Plaque Reduction Neutralization-Horseradish Peroxidase (µPRN-HP) test was used. Antibody titers were expressed as the reciprocal of the highest dilution that neutralized the challenge virus by 50 % (µPRN50). Seropositivity cut-off was set at ≥ 1:100. We included 153 participants: 46 ND-CKD, 50 D-CKD and 57 healthy adults. Median ages were 58.3, 55 and 52.2 years, respectively. Median time since YF vaccination was 22.3, 18.5 and 48.3 months respectively. There were no statistically significant differences in YF seroprevalence and neutralizing antibodies titers among groups: 100 % of ND-CKD; 96 % of D-CKD and 100 % of healthy participants were seropositive. Geometric mean titers (GMT) were 818.5, 683.0 and 665.5, respectively (p = 0.289).

Quantifying terpenes in tomato leaf extracts from different species using gas chromatography-mass spectrometry (GC-MS).

Terpenes play a vital role in plant defense; tomato plants produce a diverse range of terpenes within specialized glandular trichomes, influencing interactions with herbivores, predators, and pollinators. This study employed two distinct methods, namely leaf dip and maceration, to extract trichomes from tomato leaves. Terpene quantification was carried out using Gas Chromatography-Mass Spectrometry (GC-MS). The leaf dip method proved effective in selectively targeting trichome content, revealing unique extraction patterns compared to maceration. The GC-MS method demonstrated high linearity, accuracy, sensitivity, and low limits of detection and quantification. Application of the method to different tomato species (Solanum pennellii, Solanum pimpinellifolium, Solanum galapagense, Solanum habrochaites, and Solanum lycopersicum) identified significant variation in terpene content among these species, highlighting the potential of specific accessions for breeding programs. Notably, the terpene α-zingiberene, known for its repellency against whiteflies, was found in high quantities (211.90-9155.13 µg g-1) in Solanum habrochaites accession PI209978. These findings provide valuable insights into terpenoid diversity for plant defense mechanisms, guiding future research on developing pest-resistant tomato cultivars. Additionally, the study underscores the broader applications of terpenes in agriculture.

First-in-human demonstration of floating EMG sensors and stimulators wirelessly powered and operated by volume conduction.

BACKGROUND: Recently we reported the design and evaluation of floating semi-implantable devices that receive power from and bidirectionally communicate with an external system using coupling by volume conduction. The approach, of which the semi-implantable devices are proof-of-concept prototypes, may overcome some limitations presented by existing neuroprostheses, especially those related to implant size and deployment, as the implants avoid bulky components and can be developed as threadlike devices. Here, it is reported the first-in-human acute demonstration of these devices for electromyography (EMG) sensing and electrical stimulation. METHODS: A proof-of-concept device, consisting of implantable thin-film electrodes and a nonimplantable miniature electronic circuit connected to them, was deployed in the upper or lower limb of six healthy participants. Two external electrodes were strapped around the limb and were connected to the external system which delivered high frequency current bursts. Within these bursts, 13 commands were modulated to communicate with the implant. RESULTS: Four devices were deployed in the biceps brachii and the gastrocnemius medialis muscles, and the external system was able to power and communicate with them. Limitations regarding insertion and communication speed are reported. Sensing and stimulation parameters were configured from the external system. In one participant, electrical stimulation and EMG acquisition assays were performed, demonstrating the feasibility of the approach to power and communicate with the floating device. CONCLUSIONS: This is the first-in-human demonstration of EMG sensors and electrical stimulators powered and operated by volume conduction. These proof-of-concept devices can be miniaturized using current microelectronic technologies, enabling fully implantable networked neuroprosthetics.

Fatigue behavior of multilayer ceramic structures in traditional and reverse layering designs.

PURPOSE: This study evaluated the fatigue failure load (FFL) and the number of cycles for fatigue failure (CFF) of traditional (porcelain layer up) and reversed (zirconia layer up) designs of porcelain-veneered zirconia samples prepared with heat-pressing or file-splitting techniques. MATERIALS AND METHODS: Zirconia discs were prepared and veneered with heat-pressed or machined feldspathic ceramic. The bilayer discs were bonded onto a dentin-analog according to the bilayer technique and sample design: traditional heat-pressing (T-HP), reversed heat-pressing (R-HP), traditional file-splitting with fusion ceramic (T-FC), reversed file-splitting with fusion ceramic R-FC), traditional file-splitting with resin cement (T-RC), and reversed file-splitting with resin cement (R-RC). The fatigue tests were performed using the stepwise approach at 20 Hz, 10,000 cycles/step, step-size of 200 N starting at 600 N, and proceeding until failure detection or up to 2600 N if enduring. The failure modes (from radial and/or cone cracks) were analyzed in a stereomicroscope. RESULTS: The reversed design decreased the FFL and CFF of bilayers prepared with heat-pressing and file-splitting with fusion ceramic. The T-HP and T-FC reached the highest results, which were statistically similar between them. The bilayers prepared by the file-splitting with resin cement (T-RC and R-RC) were similar to the R-FC and R-HP groups regarding FFL and CFF. Almost all reverse layering samples failed by radial cracks. CONCLUSIONS: The reverse layering design did not improve the fatigue behavior of porcelain veneered zirconia samples. The three bilayer techniques behaved similarly when used in the reversed design.

Natural habitat connectivity and organic management modulate pest dispersal, gene flow, and natural enemy communities.

The simplification and fragmentation of agricultural landscapes generate effects on insects at multiple spatial scales. As each functional group perceives and uses the habitat differently, the response of pest insects and their associated natural enemies to environmental changes varies. Therefore, landscape structure may have consequences on gene flow among pest populations in space. This study aimed to evaluate the effects of local and landscape factors, at multiple scales, on the local infestation, gene flow and broad dispersion dynamics of the pest insect Bemisia tabaci (Genn.) Middle East-Asia Minor 1 (MEAM-1, former biotype B) (Hemiptera: Aleyrodidae) and its associated natural enemies in a tropical agroecosystem. We evaluated the abundance of B. tabaci populations and their natural enemy community in 20 tomato farms in Brazil and the gene flow between farms from 2019 to 2021. Landscapes dominated by agriculture resulted in larger B. tabaci populations and higher gene flow, especially in conventional farms. A higher density of native vegetation patches disfavored pest populations, regardless of the management system. The results revealed that whitefly responds to intermediate spatial scales and that landscape factors interact with management systems to modulate whitefly populations on focal farms. Conversely, whitefly natural enemies benefited from higher amounts of natural vegetation at small spatial scales, while the connectivity between natural habitat patches was beneficial for natural enemies regardless of the distance from the focal farm. The resulting dispersion model predicts that the movement of whiteflies between farms increases as the amount of natural vegetation decreases. Our findings demonstrate that landscape features, notably landscape configuration, can mediate infestation episodes, as they affect pest insects and natural enemies in opposite ways. We also showed that landscape features interact with farm traits, which highlights the need for management strategies at multiple spatial scales. In conclusion, we demonstrated the importance of the conservation of natural areas as a key strategy for area-wide ecological pest management and the relevance of organic farming to benefit natural enemy communities in tropical agroecosystems.

Boron-containing coating yields enhanced antimicrobial and mechanical effects on translucent zirconia.

OBJECTIVES: To evaluate the mechanical and antimicrobial properties of boron-containing coating on translucent zirconia (5Y-PSZ). METHODS: 5Y-PSZ discs (Control) were coated with a glaze (Glaze), silver- (AgCoat), or boron-containing (BCoat) glasses. The coatings' antimicrobial potential was characterized using S. mutans biofilms after 48 h via viable colony-forming units (CFU), metabolic activity (CV) assays, and quantification of extracellular polysaccharide matrix (EPS). Biofilm architectures were imaged under scanning electron and confocal laser scanning microscopies (SEM and CLSM). The cytocompatibility was determined at 24 h via WST-1 and LIVE&DEAD assays using periodontal ligament stem cells (PDLSCs). The coatings' effects on properties were characterized by Vickers hardness, biaxial bending tests, and fractography analysis. Statistical analyses were performed via one-way ANOVA, Tukey's tests, Weibull analysis, and Pearson's correlation analysis. RESULTS: BCoat significantly decreased biofilm formation, having the lowest CFU and metabolic activity compared with the other groups. BCoat and AgCoat presented the lowest EPS, followed by Glaze and Control. SEM and CLSM images revealed that the biofilms on BCoat were thin and sparse, with lower biovolume. In contrast, the other groups yielded robust biofilms with higher biovolume. The cytocompatibility was similar in all groups. BCoat, AgCoat, and Glaze also presented similar hardness and were significantly lower than Control. BCoat had the highest flexural strength, characteristic strength and Weibull parameters (σF: 625 MPa; σ0: 620 MPa; m = 11.5), followed by AgCoat (σF: 464 MPa; σ0: 478 MPa; m = 5.3). SIGNIFICANCE: BCoat is a cytocompatible coating with promising antimicrobial properties that can improve the mechanical properties and reliability of 5Y-PSZ.

Effects of solvent type and UV post-cure time on 3D-printed restorative polymers.

OBJECTIVES: This study evaluated the impact of different solvents and UV post-curing times on properties of 3D printing resins for provisional restorations. METHODS: The post-processing methods were tested using two solvents (isopropyl alcohol or absolute ethanol) and three UV times (5, 10, or 30 min). The resins tested were Resilab 3D Temp, Printax Temp, and Prizma Bioprov. Microhardness (kgf/mm2), fracture toughness (KIC, MPa√m), surface roughness (Ra, µm), gloss (gloss units), and degree of CC conversion (%DC) were measured (n = 8). All response variables were collected from the same specimen. The specimens were 3D printed using an SLA/LCD printer (150° angulation, 50 µm layer thickness). Light exposure times were adjusted for each material, and the post-processing methods were applied using an all-in-one machine immediately after printing. Data were analyzed using Three-Way ANOVA (α = 0.05). RESULTS: Microhardness was affected by UV post-cure time and 3D resin. Resilab showed higher microhardness with isopropyl alcohol and 30-min UV time, while Printax had higher microhardness with absolute ethanol. KIC was influenced by solvent type, UV time, and 3D resin, with varying effects on different resins. Roughness was affected by 3D resin and UV time, but no significant differences were seen for Resilab or Prizma. Gloss was influenced by 3D resin, and for Prizma, it was lower with specific solvent/UV time combinations. DC was influenced by 3D resin, with each resin behaving differently. SIGNIFICANCE: Tailoring the combination of 3D resin, solvent washing type, and UV post-curing time is important to achieve optimal mechanical and aesthetic outcomes for restorations.

Borosilicate glass as a surface finishing alternative for improving the mechanical properties of third-generation zirconia.

OBJECTIVE: This study evaluated the effect of an experimental borosilicate glass on the mechanical and optical behavior of 5Y-PSZ zirconia and comparing it to commercial glaze and as-sintered. METHODS: Disc-shaped specimens of a 5Y-PSZ (Zpex Smile) were prepared and sintered (1550 °C, 2 h). The zirconia discs were randomly divided according to the surface treatment: as-sintered (C), commercial glaze (G), and experimental borosilicate glass (SL). Glaze and experimental glass powders were mixed with building liquids and applied to zirconia with a brush. G specimens were fired at 950 °C and SL at 1200 °C. An extended dwell time of 20 min was applied to both groups. Biaxial flexural strength, roughness (Ra and Rz), translucency (TP00), color alteration (ΔE00), Vickers hardness, fracture toughness, residual stresses, and x-ray diffraction analyses were conducted. Statistical analyses were performed with Weibull statistics, Kruskal-Wallis, or ANOVA tests (α = 5%). RESULTS: SL yielded the highest flexural strength (799.35 MPa), followed by G (662.34 MPa), and C (485.38 MPa). The fracture origin of SL specimens was in the bulk zirconia, while G and C showed fractures starting at the surface. As-sintered reached the highest fracture toughness and hardness. Glaze and borosilicate glass provided surface compressive stresses. Borosilicate glass application led to phase transformation (t→m). SL and G showed the lowest roughness. TP00 and ΔE00 were similar among groups. SIGNIFICANCE: Borosilicate glass improved strength without harming the optical properties of third-generation zirconia. Toughness and roughness provided by the experimental glass were similar to those from commercial glaze.

Association between PAH and plastic fragments on Brazilian coast beaches: a baseline assessment.

Plastic materials have many destinies on the environment, and one of them is its continuous fragmentation, originating microplastics (MP). MP act as an adsorption surface for organic pollutants like polycyclic aromatic hydrocarbons (PAH). Due to its toxic and harmful properties, 16 PAH are defined as priorities in studies. Several works have evaluated the occurrence of MP and PAH individually in environmental matrices, but just a few have investigated their association. The present study aims to investigate PAH occurrence, sources, and concentration in MP from beach sediments of eight Brazilian coastal states. Sixteen PAH were analyzed by gas chromatography, in which 14 PAH were detected, and PAH total concentration (ΣPAH) ranged from 0.25 to 71.60 ng g-1. In the North and Northeast regions, the low PAH concentrations in MP appear to be related to the intense hydrodynamic processes. Naphthalene levels on the Styrofoam were above the threshold effect level in two samples and near it on one; therefore, effects in the organisms might occur due to these PAH. The Southeast and South regions are the only ones with previous studies relating PAH with MP, and ΣPAH ranged from 0.34 to 2.52 ng g-1. According to the diagnostic ratios, the PAH sources were associated with industrial and port activities, untreated effluent disposal, and urban runoff by pluvial waters. The highest ΣPAH level was found in the Styrofoam samples, suggesting that this MP type might contribute considerable to PAH dispersion on Brazilian coast beaches.

Augmented Reality Feedback for Exoskeleton-Assisted Walking. A Feasibility Study.

Powered lower limb exoskeletons have been used in recent decades to support and improve conventional gait rehabilitation programs. In this context, visual feedback is considered a valuable tool to facilitate patient learning and engagement. Treadmill-based lower limb robotic exoskeletons are commonly incorporated with traditional screens or virtual reality (VR) devices. However, these modalities are incompatible with over-ground robotic exoskeletons, in which users should pay attention to multiple elements of the open environment and more complex tasks. Recent advances in augmented reality (AR) technology have unlocked a new way of displaying 3D graphics in untethered wearable devices like Microsoft's Hololens 2 without occluding the rest of the user's field of view. These advances can be crucial in certain situations and position AR as an excellent candidate to improve visual feedback when using robotic exoskeletons. In this work, we present the development of an AR-based audio-visual feedback system that tracks the trunk rotation from an Inertial Measurement Unit (IMU) to aid in walking tasks assisted by a lower limb robotic exoskeleton. A preliminary study was done integrating the H3 robotic exoskeleton with Hololens 2. We observed that displaying posture information is feasible and could help mitigate the unnatural posture often imposed by a robotic exoskeleton.

Molecular mechanism of glutaminase activation through filamentation and the role of filaments in mitophagy protection.

Glutaminase (GLS), which deaminates glutamine to form glutamate, is a mitochondrial tetrameric protein complex. Although inorganic phosphate (Pi) is known to promote GLS filamentation and activation, the molecular basis of this mechanism is unknown. Here we aimed to determine the molecular mechanism of Pi-induced mouse GLS filamentation and its impact on mitochondrial physiology. Single-particle cryogenic electron microscopy revealed an allosteric mechanism in which Pi binding at the tetramer interface and the activation loop is coupled to direct nucleophile activation at the active site. The active conformation is prone to enzyme filamentation. Notably, human GLS filaments form inside tubulated mitochondria following glutamine withdrawal, as shown by in situ cryo-electron tomography of cells thinned by cryo-focused ion beam milling. Mitochondria with GLS filaments exhibit increased protection from mitophagy. We reveal roles of filamentous GLS in mitochondrial morphology and recycling.

A clinical model to predict successful renal replacement therapy (RRT) discontinuation in patients with Acute Kidney Injury (AKI).

INTRODUCTION: Ideal timing of Renal Replacement Therapy (RRT) discontinuation in Acute Kidney Injury (AKI) is still unknown. We aimed to study the role of creatinine-related variables in predicting RRT successful discontinuation and to propose a clinical predictive score. METHODS: In this single-centre retrospective study, we evaluated all AKI patients in whom RRT was interrupted for at least 48 hours. Patients who were still RRT-independent 7 days after initial RRT cessation were included in the "Success" group and opposed to the "Failure" group. We evaluated baseline characteristics and variables collected at the time of RRT interruption, as well as the Kinetic estimated Glomerular Filtration Rate (KeGFR), the simple variation in serum Creatinine (ΔsCr), and the incremental creatinine ratio on the first three days after RRT interruption. Multivariable analysis was performed to evaluate prediction of success. Internal validation using a simple binomial generalized regression model with Lasso estimation and 5-fold cross validation method was performed. RESULTS: We included 124 patients, 49 in the "Failure" group and 75 in the "Success" group. All creatinine-related variables predicted success in simple and multiple logistic regression models. The best model generated a clinical score based on the odds ratio obtained for each variable and included urine output, non-renal SOFA score, fluid balance, serum urea, serum potassium, blood pH, and the variation in sCr values after RRT discontinuation. The score presented an area under the ROC of 0.86 (95% CI 0.76‒1.00). CONCLUSION: Creatinine variation between the first 2 consecutive days after RRT discontinuation might predict success in RRT discontinuation. The developed clinical score based on these variables might be a useful clinical decision tool to guide hemodialysis catheter safe removal.

Bonding stability of universal adhesives to feldspathic ceramic with or without prior silane application.

PURPOSE: This study aimed to assess the bonding stability of three universal adhesives to a CAD-CAM feldspathic ceramic with or without prior silane application. MATERIALS AND METHODS: The universal adhesives tested were Ambar Universal (FGM), Single Bond Universal (3M), and Ybond Universal (Yller). Scotchbond Multipurpose Plus (3M) was used as a control. The silane used was RelyX Ceramic Primer (3M). Microtensile bond strength (µTBS) to Vitablocs Mark II was measured after 24 h or 6 months of water aging (n = 20). Failure modes were observed through scanning electron microscopy, and pH and degree of C=C conversion (DC) of the adhesives were measured (n = 3). Data were analyzed using one-, two-, and three-way ANOVA. RESULTS: The pH values differed significantly among the adhesives (p < 0.001), while the DC was similar (p = 0.141). The universal adhesives showed significant differences in µTBS, with higher immediate results when using silane and lower results after aging. Notably, a significant decrease in µTBS was observed after 6 months when silane was applied. In contrast, the control adhesive exhibited consistent results between the 24-h and 6-month storage periods. The universal adhesives showed a decrease in µTBS ranging from 25.5% to 40.1% after prolonged storage. Adhesive failures were predominant in all groups. The presence or absence of silane in the adhesive composition did not affect the bonding performance. CONCLUSIONS: The application of a separate silane coupling agent improved the immediate bond strength of universal adhesives to CAD-CAM feldspathic ceramic. However, this bond strength significantly decreased after 6 months of water aging. The bond strength remained stable for universal adhesives when applied without silane after 6 months of aging.

Interface adhesion on layered zirconia: Effects of the veneering ceramic material and veneering technique.

PURPOSE: To evaluate the effect of different veneering ceramics and veneering techniques on the bond strength to zirconia. MATERIALS AND METHODS: 3Y-TZP zirconia blocks were sliced into 60 slabs, polished, and sintered. Each slab received one ceramic cylinder (Ø = 3.4 mm, 5 mm-high), according to the veneering ceramic type (feldspathic-FEL or lithium disilicate-based-LD) and the veneering technique (file-splitting with resin-based luting agent-RC, file-splitting with fusion ceramic-FC, or heat-pressing-HT), which resulted in six groups: FEL-RC, FEL-FC, FEL-HT, LD-RC, LD-FC, LD-HT. After preparation, the samples were immersed in distilled water for 24 h before the shear bond strength (SBS) test. The failure modes were classified as adhesive, predominantly adhesive, or cohesive. Representative failure mode images were taken in a Scanning Electron Microscope. The SBS data were analyzed by two-way ANOVA and Tukey's test. RESULTS: Both type of veneering ceramic and technique affected the bond strength. FC led to the highest SBS values. RC and HP provided similar results when compared within each veneering ceramic. Lithium disilicate achieved lower bond strength than feldspathic ceramic when the heat-pressing technique was applied. The most frequent failure modes were predominantly adhesive and adhesive for FEL and LD, respectively. CONCLUSION: File-splitting with fusion ceramic provided the highest adhesion to zirconia when feldspathic or lithium disilicate-based ceramics were used. The heat-pressing technique for veneering with lithium disilicate significantly decreased the bond strength when compared to the feldspathic ceramic.

TREATMENT WITH HUMAN UMBILICAL CORD-DERIVED MESENCHYMAL STEM CELLS IN A PIG MODEL OF SEPSIS-INDUCED ACUTE KIDNEY INJURY: EFFECTS ON MICROVASCULAR ENDOTHELIAL CELLS AND TUBULAR CELLS IN THE KIDNEY.

ABSTRACT: Background: Approximately 50% of patients with sepsis develop acute kidney injury (AKI), which is predictive of poor outcomes, with mortality rates of up to 70%. The endothelium is a major target for treatments aimed at preventing the complications of sepsis. We hypothesized that human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) could attenuate tubular and endothelial injury in a porcine model of sepsis-induced AKI. Methods: Anesthetized pigs were induced to fecal peritonitis, resulting in septic shock, and were randomized to treatment with fluids, vasopressors, and antibiotics (sepsis group; n = 11) or to that same treatment plus infusion of 1 × 10 6 cells/kg of hUC-MSCs (sepsis+MSC group; n = 11). Results: At 24 h after sepsis induction, changes in serum creatinine and mean arterial pressure were comparable between the two groups, as was mortality. However, the sepsis+MSC group showed some significant differences in comparison with the sepsis group: lower fractional excretions of sodium and potassium; greater epithelial sodium channel protein expression; and lower protein expression of the Na-K-2Cl cotransporter and aquaporin 2 in the renal medulla. Expression of P-selectin, thrombomodulin, and vascular endothelial growth factor was significantly lower in the sepsis+MSC group than in the sepsis group, whereas that of Toll-like receptor 4 (TLR4) and nuclear factor-kappa B (NF-κB) was lower in the former. Conclusion: Treatment with hUC-MSCs seems to protect endothelial and tubular cells in sepsis-induced AKI, possibly via the TLR4/NF-κB signaling pathway. Therefore, it might be an effective treatment for sepsis-induced AKI.

Precision management of acute kidney injury in the intensive care unit: current state of the art.

Acute kidney injury (AKI) is a prototypical example of a common syndrome in critical illness defined by consensus. The consensus definition for AKI, traditionally defined using only serum creatinine and urine output, was needed to standardize the description for epidemiology and to harmonize eligibility for clinical trials. However, AKI is not a simple disease, but rather a complex and multi-factorial syndrome characterized by a wide spectrum of pathobiology. AKI is now recognized to be comprised of numerous sub-phenotypes that can be discriminated through shared features such as etiology, prognosis, or common pathobiological mechanisms of injury and damage. The characterization of sub-phenotypes can serve to enable prognostic enrichment (i.e., identify subsets of patients more likely to share an outcome of interest) and predictive enrichment (identify subsets of patients more likely to respond favorably to a given therapy). Existing and emerging biomarkers will aid in discriminating sub-phenotypes of AKI, facilitate expansion of diagnostic criteria, and be leveraged to realize personalized approaches to management, particularly for recognizing treatment-responsive mechanisms (i.e., endotypes) and targets for intervention (i.e., treatable traits). Specific biomarkers (e.g., serum renin; olfactomedin 4 (OLFM4); interleukin (IL)-9) may further enable identification of pathobiological mechanisms to serve as treatment targets. However, even non-specific biomarkers of kidney injury (e.g., neutrophil gelatinase-associated lipocalin, NGAL; [tissue inhibitor of metalloproteinases 2, TIMP2]·[insulin like growth factor binding protein 7, IGFBP7]; kidney injury molecule 1, KIM-1) can direct greater precision management for specific sub-phenotypes of AKI. This review will summarize these evolving concepts and recent innovations in precision medicine approaches to the syndrome of AKI in critical illness, along with providing examples of how they can be leveraged to guide patient care.