Mitochondrial DNA levels in perfusate and bile during ex vivo normothermic machine correspond with donor liver quality.

Ex vivo normothermic machine perfusion (NMP) preserves donor organs and permits real-time assessment of allograft health, but the most effective indicators of graft viability are uncertain. Mitochondrial DNA (mtDNA), released consequent to traumatic cell injury and death, including the ischemia-reperfusion injury inherent in transplantation, may meet the need for a biomarker in this context. We describe a real time PCR-based approach to assess cell-free mtDNA during NMP as a universal biomarker of allograft quality. Measured in the perfusate fluid of 29 livers, the quantity of mtDNA correlated with metrics of donor liver health including International Normalized Ratio (INR), lactate, and warm ischemia time, and inversely correlated with inferior vena cava (IVC) flow during perfusion. Our findings endorse mtDNA as a simple and rapidly measured feature that can inform donor liver health, opening the possibility to better assess livers acquired from extended criteria donors to improve organ supply.

In vitro evaluation of the osteogenic and antimicrobial potential of porous wollastonite scaffolds impregnated with ethanolic extracts of propolis.

Context: The development of porous devices using materials modified with various natural agents has become a priority for bone healing processes in the oral and maxillofacial field. There must be a balance between the proliferation of eukaryotic and the inhibition of prokaryotic cells to achieve proper bone health. Infections might inhibit the formation of new alveolar bone during bone graft augmentation. Objective: This study aimed to evaluate the in vitro osteogenic behavior of human bone marrow stem cells and assess the antimicrobial response to 3D-printed porous scaffolds using propolis-modified wollastonite. Methodology: A fractional factorial design of experiments was used to obtain a 3D printing paste for developing scaffolds with a triply periodic minimal surface (TPMS) gyroid geometry based on wollastonite and modified with an ethanolic propolis extract. The antioxidant activity of the extracts was characterized using free radical scavenging methods (DPPH and ABTS). Cell proliferation and osteogenic potential using Human Bone Marrow Stem Cells (bmMSCs) were assessed at different culture time points up to 28 days. MIC and inhibition zones were studied from single strain cultures, and biofilm formation was evaluated on the scaffolds under co-culture conditions. The mechanical strength of the scaffolds was evaluated. Results: Through statistical design of experiments, a paste suitable for printing scaffolds with the desired geometry was obtained. Propolis extracts modifying the TPMS gyroid scaffolds showed favorable cell proliferation and metabolic activity with osteogenic potential after 21 days. Additionally, propolis exhibited antioxidant activity, which may be related to the antimicrobial effectiveness of the scaffolds against S. aureus and S. epidermidis cultures. The mechanical properties of the scaffolds were not affected by propolis impregnation. Conclusion: These results demonstrate that propolis-impregnated porous wollastonite scaffolds might have the potential to stimulate bone repair in maxillofacial tissue engineering applications.

Microplásticos en la sardina Sardinella aurita (Teleostei: Clupeidae) en Sucre, Venezuela / Microplastics in the sardine Sardinella aurita (Teleostei: Clupeidae) in Sucre, Venezuela

Resumen Introducción: La creciente presencia de plásticos en los ecosistemas acuáticos ha sido considerada como un problema ambiental emergente global. Existen estudios que han demostrado que los microplásticos pueden ser ingeridos por una variedad de organismos acuáticos. La variabilidad natural e importancia del recurso sardina, son razones suficientes para evaluar la presencia de estas partículas en el contenido estomacal, y así generar información acerca de las implicaciones de ingestión de estos contaminantes emergentes. Objetivo: Evaluar la presencia de microplásticos en el contenido estomacal de la sardina y su correlación con la condición fisiológica y reproductiva de los peces estudiados. Metodología: Se evaluaron un total de 800 sardinas, entre abril y mayo del 2022. Se midieron las variables morfométricas longitud total y peso, se determinó el factor de condición y el índice gonadosomático. Se extrajo el tracto digestivo y se determinó el peso del estómago lleno y vacío, además del peso del contenido estomacal para determinar el índice de repleción y vacuidad. Las variables determinadas se correlacionaron con la abundancia de microplásticos. El contenido estomacal fue procesado para extraer los microplásticos y caracterizarlos de forma física. Resultados: El índice de repleción presentó diferencias mensuales; su promedio fue de 0.311. El índice de vacuidad promedio fue de 0.276. La abundancia de microplásticos promedio fue 3 066 ítems, con diferencias mensuales e interacción con respecto al sexo y una frecuencia de aparición de 70.125 %, con un total de 2 402 fibras y 57 fragmentos. Las fibras negras fueron las más abundantes (947 ítems). Encontramos correlaciones bajas y positivas entre la abundancia de microplásticos, el índice gonadosomático y el peso de los organismos. Conclusiones: Estos resultados no pueden ser concluyente con respecto a las consecuencias causadas por este contaminante emergente. Mas estudios son necesarios, incluyendo ensayos de laboratorios con metodologías estandarizadas.

Abstract Introduction: The increasing presence of plastics in aquatic ecosystems has been considered as an emerging global environmental problem. Studies have shown that microplastics can be ingested by a variety of aquatic organisms. The natural variability and importance of the sardine resource are sufficient reasons to evaluate the presence of these particles in stomach contents, and thus generate information about the ingestion implications of these emerging contaminants. Objective: To evaluate the presenc e of microplastics in the stomach contents of sardines and their correlation with the physiological and reproductive condition of the fish studied. Methodology: We evaluated a total of 800 sardines between April and May 2022. We measured total length and weight, and determined the condition factor and the gonadosomatic index. We removed the digestive tract and weighed the stomach both empty and full, as well as the stomach contents to determine the repletion and emptiness index. We correlated the determined variables with the abundance of microplastics. We extracted microplastics and characterized them physically. Results: The filling index presented monthly differences with an average of 0.311. The average vacancy index was 0.276. The average abundance of microplastics was 3 066 items, with monthly differences and interaction with respect to sex and a frequency of appearance of 70.125 %, with a total of 2 402 fibers and 57 fragments. Black fibers were the most abundant (947 items). Low and positive correlations were found between the abundance of microplastics, the gonadosomatic index and the weight of the organisms. Conclusions: These results cannot be conclusive as to the consequences caused by this emerging pollutant. Further study is necessary, including laboratory tests with standardized methodologies.

Chemical Solution Deposition of Protective Er2O3 and Y2O3 Coatings onto Stainless Steel for Molten Metal Casting using Metal-Nitrate Precursors.

Chemical solution deposition (CSD) methods involving the thermal decomposition of 5.0 M Er(NO3)3·5H2O and Y(NO3)3·6H2O precursor solutions were employed to fabricate protective erbia and yttria coatings onto stainless steel (SS304/SS316) coupons. The two techniques tested were dip and spray coating, which were then compared to a commercial yttria spray (ZYP Coatings). It was determined that solution concentration, solvent choice, injection of Er2O3 and Y2O3 micropowder, and the annealing temperature/ramp profile were critical to the coatings' physical properties. For dip coatings, thicknesses were 1-20 µm after two dipping/annealing cycles, and adhesion strength was ∼1000 psi, increasing up to ∼1300 psi if the SS coupons had preliminary sandblasting. Spray coatings from precursor solutions were reported to have thicknesses of 20-80 µm and adhesion strength less than 500 psi (regardless of the coupon surface finish). Cross-sectional views of the coatings confirmed subsurface porosity, and XRD results indicated that the coatings were polycrystalline, with patterns typical to that of cubic Er2O3 and Y2O3.

Roles of HNO x and Carboxylic Acids in the Thermal Stability of Nitroplasticizer.

In the thermal aging of nitroplasticizer (NP), the produced nitrous acid (HONO) can decompose into reactive nitro-oxide species and nitric acid (HNO3). These volatile species are prone to cause cascaded deterioration of NP and give rise to various acidic constituents. To gain insight on the early stage of NP degradation, an adequate method for measuring changes in the concentrations of HONO, HNO3, and related acidic species is imperative. The typical assessment of acidity in nonaqueous solutions (i.e., acid number) cannot differentiate acidic species and thus presents difficulty in the measurement of HONO and HNO3 at a micromolar concentration level. Using liquid-liquid extraction and ion chromatography (IC), we developed a fast and unambiguous analytical method to accurately determine the concentration of HONO, HNO3, acetic/formic acids, and oxalic acid in aged NP samples. Given by the overlay analysis results of liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and IC, the prominent increase of produced HONO after the depletion of antioxidants is the primary cause of HNO3 formation in the late stage of NP degradation, which results in the acid-catalyzed hydrolysis of NP into 2,2-dinitropropanol and acetic/formic acids. Our study has demonstrated that the aging temperature plays a crucial role in accelerating the formation and decomposition of HONO, which consequently increases the acidity of aged NP samples and hence accelerates the hydrolyzation of NP. Therefore, to prevent NP from undergoing rapid degradation, we suggest that the concentration of HNO3 should be maintained below 1.35 mM and the temperature under 38 °C.

Hernia diafragmática traumática derecha en paciente geriátrico. Reporte de un caso infrecuente / Traumatic right diaphragmatic hernia in an elderly patient. A rare case report

Introducción: La hernia diafragmática surge como resultado del movimiento de los órganos abdominales hacia el tórax a través de un defecto del diafragma resultante de varios tipos de lesiones, que a menudo representan un desafío diagnóstico y terapéutico. La hernia diafragmática traumática es una enfermedad infrecuente. El trauma diafragmático rara vez es aislado, la mayoría de las veces se acompaña de otras lesiones toracoabdominales, cerebrales o musculoesqueléticas, siendo estas comorbilidades más graves responsables del mal pronóstico y aumento de la mortalidad. Caso clínico: Anciana de 90 años de edad, hipertensa, quien inicia cuadro clínico 48 horas previas a su ingreso, cuando posterior a caída del plano de bipedestación presenta disminución del estado de consciencia, náuseas, vómitos y disnea en reposo. Rx de tórax evidencia asas intestinales en hemitórax derecho. Se realizó laparotomía exploradora. Conclusión: Las hernias diafragmáticas traumáticas son infrecuentes, con incidencia <3%, sintomatología inespecífica y relacionadas generalmente con lesiones asociadas al trauma. Siempre se debe sospechar de una HD ante un paciente con antecedente de trauma cerrado de alto impacto o penetrante, en función de la cinética y mecanismo de lesión. El diagnóstico supone un reto para el cirujano, debiendo apoyarse en una firme sospecha y estudios radiológicos. Su tratamiento es quirúrgico, debiendo ser individualizado, con abordajes torácicos o abdominales en función del caso. La técnica a emplear dependerá de las características del defecto, de la fase del diagnóstico y la experiencia del equipo quirúrgico(AU)

Introduction: Diaphragmatic hernia arises as a result of the movement of the abdominal organs towards the thorax through a defect in the diaphragm resulting from various types of injuries, which often represent a diagnostic and therapeutic challenge. Traumatic diaphragmatic hernia is a rare disease. Diaphragmatic trauma is rarely isolated, most of the time it is accompanied by other thoracoabdominal, brain or musculoskeletal injuries, these more serious comorbidities being responsible for the poor prognosis and increased mortality. Clinical case: A 90-year-old woman with hypertension , who began a clinical picture 48 hours prior to admission, when after falling from the standing plane he presented a decreased state of consciousness, nausea, vomiting, and dyspnea at rest. Chest x-ray shows intestinal loops in the right hemithorax. Exploratory laparotomy was performed. Conclusion: Traumatic diaphragmatic hernias are rare, with an incidence of <3%, non-specific symptoms, and generally related to injuries associated with trauma. HD should always be suspected in a patient with a history of high-impact or penetrating blunt trauma, depending on the kinetics and mechanism of injury. Diagnosis is a challenge for the surgeon, and must be based on a firm suspicion and radiological studies. Its treatment is surgical, and must be individualized, with thoracic or abdominal approaches depending on the case. The technique to be used will depend on the characteristics of the defect, the diagnostic phase and the experience of the surgical team(AU)

Structure, Process, and Mortality Associated with Acute Coronary Syndrome Management in Guatemala's National Healthcare System: The ACS-GT Registry.

Background: Acute coronary syndromes (ACS) include ST-segment elevation myocardial infarction (STEMI), non-ST-segment elevation myocardial infarction (NSTEMI), and unstable angina (UA). The leading cause of mortality in Guatemala is acute myocardial infarction (AMI) and there is no established national policy nor current standard of care. Objective: Describe the factors that influence ACS outcome, evaluating the national healthcare system's quality of care based on the Donabedian health model. Methods: The ACS-Gt study is an observational, multicentre, and prospective national registry. A total of 109 ACS adult patients admitted at six hospitals from Guatemala's National Healthcare System were included. These represent six out of the country's eight geographic regions. Data enrolment took place from February 2020 to January 2021. Data was assessed using chi-square test, Student's t-test, or Mann-Whitney U test, whichever applied. A p-value < 0.05 was considered statistically significant. Results: One hundred and nine patients met inclusion criteria (80.7% STEMI, 19.3% NSTEMI/UA). The population was predominantly male, (68%) hypertensive (49.5%), and diabetic (45.9%). Fifty-nine percent of STEMI patients received fibrinolysis (alteplase 65.4%) and none for primary Percutaneous Coronary Intervention (pPCI). Reperfusion success rate was 65%, and none were taken to PCI afterwards in the recommended time period (2-24 hours). Prognostic delays in STEMI were significantly prolonged in comparison with European guidelines goals. Optimal in-hospital medical therapy was 8.3%, and in-hospital mortality was 20.4%. Conclusions: There is poor access to ACS pharmacological treatment, low reperfusion rate, and no primary, urgent, or rescue PCI available. No patient fulfilled the recommended time period between successful fibrinolysis and PCI. Resources are limited and inefficiently used.

Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry Analysis of Eutectic Bis(2,2-dinitropropyl) Acetal/Formal Degradation Profile: Nontargeted Identification of Antioxidant Derivatives.

In the eutectic mixture of bis(2,2-dinitropropyl) acetal (BDNPA) and bis(2,2-dinitropropyl) formal (BDNPF), also known as nitroplasticizer (NP), n-phenyl-ß-naphthylamine (PBNA), an antioxidant, is used to improve the long-term storage of NP. PBNA scavenges nitrogen oxides (e.g., NO x radicals) that are evolved from NP decomposition, hence slowing down the degradation of NP. Yet, little is known about the associated chemical reaction between NP and PBNA. Herein, using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF), we thoroughly characterize nitrated PBNA derivatives with up to five NO2 moieties in terms of retention time, mass verification, fragmentation pattern, and correlation with NP degradation. The propagation of PBNA nitration is found to depend on the temperature and acidity of the NP system and can be utilized as an indirect, yet reliable, means of determining the extent of NP degradation. At low temperatures (<55 °C), we find that the scavenging efficiency of PBNA is nullified when three NO2 moieties are added to PBNA. Hence, the dinitro derivative can be used as a reliable indicator for the onset of hydrolytic NP degradation. At elevated temperatures (≥55 °C) and especially in the dry environment, the trace amount of water in the condensed NP (<700 ppm) is essentially removed, which accelerates the production of reactive species (e.g., HONO, HNO3 and NO x ). In return, the increased acidity due to HNO3 formation catalyzes the hydrolysis of NP and PBNA nitro derivatives into 2,2-dinitropropanol (DNPOH) and nitrophenol/dinitrophenol, respectively.

Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry: A Strategy for Optimization, Characterization, and Quantification of Antioxidant Nitro Derivatives.

As an antioxidant, N-phenyl-ß-naphthylamine (PBNA) inhibits the activity of oxidants, such as NO x , to prevent the degradation of energetic materials. In the presence of NO x , nitrated products can be generated in the process potentially. To characterize nitrated PBNA in a nontargeted analysis of complex samples as such, liquid chromatography tandem quadrupole time-of-flight (LC-QTOF), as an excellent analytic technique, is used due to its high resolution and sensitivity. However, a systematic approach of instrumentation optimization, data interpretation, and quantitative determination of products is needed. Through a step-by-step evaluation of the instrumental parameters used in the Q0, Q1, and Q2 compartments of LC-QTOF, optimal ion yields of precursor ions and high-resolution MS2 fragmentation spectra at low mass defects were obtained in both negative and positive electrospray ionization modes. Through rationalization of the fragmentation pathways and verification using theoretical masses, the mononitro derivative of PBNA was accurately identified as N-(4-nitrophenyl)-naphthalen-2-amine and further confirmed using a reference standard. Using strict criteria provided by the analytical guidelines (e.g., SANTE), limit of quantitation, limit of detection, and calibration were established for the quantitation of PBNA and nitrated PBNA. From optimization to characterization and subsequent quantification of the mononitro-PBNA derivative, for the first time, the applicability of this strategy is demonstrated in the aged energetic binders.

Metamorphic diamond from the northeastern margin of Gondwana: Paradigm shifting implications for one of Earth's largest orogens.

We describe the first occurrence of diamond-facies ultrahigh pressure metamorphism along the Gondwana-Pacific margin of the Terra Australis Orogen. Metamorphic garnet grains from Ordovician metasediments along the Clarke River Fault in northeastern Queensland contain inclusions of diamond and quartz after coesite, as well as exsolution lamellae of rutile, apatite, amphibole, and silica. These features constrain minimum pressure-temperature conditions to >3.5 gigapascals and ~860°C, although peak pressure conditions may have exceeded 5 gigapascals. On the basis of these data, we interpret the Clarke River Fault to represent a Paleozoic suture zone and at least parts of the Terra Australis Orogen to have formed through classic Wilson cycle processes. The growth of the Terra Australis Orogen during the Paleozoic is largely attributed to accretionary style tectonics. These previously unknown findings indicate that the Terra Australis Orogen was not just a simple accretionary style orogen but rather a complex system with multiple tectonic styles operating in tandem including collisional tectonics.

Determination of structural properties in the adsorption of drugs on chitosan-hydrogels for type 2 diabetes by means of the PM6 method / Determinación de propiedades estructurales en la adsorción de fármacos en hidrogeles de quitosano para diabetes tipo 2 mediante el método PM6 / Determinação das propriedades estruturais en adsorção de drogas em hidrogelos de quitosan para diabetes do tipo 2 pelo método PM6

Abstract Computational chemistry performs the modeling and calculation of physicochemical properties that allow understanding of the different molecular interactions at the nanometric scale in medical applications such as the design of controlled release systems. The PM6 model was used to analyze metformin and glibenclamide. First, the energy properties as the Gibbs free energy and enthalpies were obtained. The results showed the affinity of both drugs with water (glibenclamide: -7.96 and metformin: -11.49) due to the formation of hydrogen bonds, which were verified by the electronegativities corresponding to the dipole moment and to the partition coefficient (Log P). Subsequently, the main properties for the design of a release system using the metformin/glibenclamide complex in the chitosan hydrogel were determined. In this process it was appreciated that the Gibbs free energy (-2157.60 kcal/ mol) determined the thermodynamic stability of the adsorption. In addition, the Log P (-25.82) indicated an instantaneous solubility through the formation of hydrogen bonds and were verified by the electronic distribution and the change in dipole moment.

Resumen La química computacional realiza el modelado y el cálculo de propiedades fisicoquímicas que permiten comprender las diferentes interacciones moleculares a escala nanométrica en aplicaciones médicas como el diseño de sistemas de liberación controlada, por ejemplo. El modelo PM6 se utilizó para analizar metformina y glibenclamida. Primero se obtuvieron las propiedades energéticas como la energía libre de Gibbs y las entalpias. Los resultados mostraron la afinidad de ambos fármacos con el agua (glibenclamide: -7,96 y metformina: -11,49) debido a la formación de enlaces de hidrógeno que fueron verificados por las electronegatividades correspondientes al momento dipolar y al coeficiente de partición (Log P). Posteriormente, se determinaron las principales propiedades para el diseño de un sistema de liberación que usa el complejo metformina/glibenclamida en el hidrogel de quitosano. En este proceso se apreció que la energía libre de Gibbs (-2157,60 kcal/mol) determinó la estabilidad termodinámica de la adsorción. Además, el Log P (-25,82) indicó una solubilidad instantánea a través de la formación de enlaces de hidrógeno y se verificó mediante la distribución electrónica y el cambio en el momento dipolar.

Resumo A química computacional realiza a modelagem e o cálculo das propriedades físico-químicas que permitem compreender as diferentes interações moleculares em escala nanométrica em aplicações médicas, como o projeto de sistemas de liberação controlada. O modelo PM6 é usado para analisar metformina e glibenclamida. Primeiro, obtenha as propriedades energéticas como a energia livre de Gibbs e as entalpias. Os resultados mostram a afinidade de ambos os componentes com água (glibenclamida: -7.96 e forma: -11.49) debitados na forma de ligações de hidrogénio, que verificam por eletronegatividades correspondentes no momento dipolar e coeficiente de participação (Log P). Posteriormente, selecione as principais propriedades para o projeto de um sistema de liberação que usa o método completo/glibenclamida no hidrogel de quitosano. Neste processo, aprecie a energia livre de Gibbs (-2157.60 kcal/mol) que determina a estabilidade termodinâmica da adsorção. Além disso, o Log P (-25.82) indica uma solução instantânea para a passagem de forma de hidrogénio e é verificada usando a distribuição eletrônica e o câmbio no momento dipolar.

Maternidad/Paternidad: el declive de roles sociales convencionales en una muestra de universitarios(as) / Maternity / Paternity: the decline of conventional social roles in a sample of university students / Maternidade / Paternidade: o declínio dos papéis sociais convencionais em uma amostra de estudiantes universitarios

Objetivo: Determinar las percepciones sobre la maternidad y paternidad de un grupo de estudiantes adolescentes y jóvenes universitarios en Santander (Colombia) en el 2019. Materiales y métodos: Estudio cualitativo descriptivo con enfoque desde la teoría fundamentada; se realizaron 22 entrevistas a jóvenes universitarios. La técnica de recolección de datos fue la entrevista semiestructurada con guión validado por expertos. El proyecto fue aprobado por el comité de ética de investigación de la Fundación Universitaria de San Gil UNISANGIL. Resultados: Gran parte de los entrevistados dan cuenta de su decisión de postergar e incluso descartar la idea de la maternidad/ paternidad de su proyecto de vida anteponiendo logros académicos y laborales; asumir el ser madre o padre durante la vida universitaria es un gran reto para los jóvenes y puede suponer un freno para aquellos que buscan mejorar su calidad de vida y superar ciclos de pobreza. Conclusiones: La investigación permitió dar cuenta de los cambios que se comienzan a gestar en las intenciones de los jóvenes de posponer o definitivamente no optar por la maternidad y /o la paternidad en sus aspiraciones vitales.

Objective: To determine the perceptions about motherhood and fatherhood of a group of adolescent students and young university students in Santander (Colombia) in 2019. Materials and methods: Descriptive qualitative study with a grounded theory approach; 22 interviews were conducted with young university students. The data collection technique was the semi-structured interview with a script validated by experts. The project was approved by the research ethics committee of the Foundation Universitaria de San Gil UNISANGIL. Results: A large part of the interviewees gave an account of their decision to postpone and even discard the idea of motherhood / fatherhood from their life project, putting academic and work achievements first; assume being a mother or father during life College is a great challenge for young people and can be a brake for those who seek to improve their quality of life and overcome cycles of poverty. Conclusions: The research allowed to give account of the changes that are beginning to take shape in the intentions of young people to postpone or definitely not choose motherhood and / or fatherhood in their life aspirations.

Objetivo: Determinar as percepções sobre a maternidade e a paternidade de um grupo de dolescentes estudantes e jovens universitários de Santander (Colômbia) em 2019. Materiais e métodos: Estudo qualitativo descritivo com abordagem da teoria fundamentada; Foram realizadas 22 entrevistas com jovens universitários. A técnica de coleta de dados foi a entrevista semiestruturada com roteiro validado por especialistas. O projeto foi aprovado pelo comitê de ética em pesquisa da Fundación Universitaria de San Gil UNISANGIL. Resultados: Grande parte das entrevistadas relatou sua decisão de adiar e até mesmo descartar a ideia de maternidade / paternidade de seu projeto de vida, priorizando as conquistas acadêmicas e profissionais; assumir o ser pai ou mãe durante a vida. A faculdade é um grande desafio para os jovens e pode ser um freio para aqueles que buscam melhorar sua qualidade de vida e superar ciclos de pobreza. Conclusões: A pesquisa permitiu dar conta as mudanças que começam a se delinear nas intenções dos jovens de adiar ou definitivamente não optar pela maternidade e / ou paternidade em suas aspirações de vida.

Effect of Conditioning Protocols and Ultrasonic Application of an Infiltrant Resin in White Spot Lesions.

The aim of this study was to evaluate different conditioning protocols and sonic/ultrasonic application of an infiltrant resin (IR) in artificial white spot lesions (AWSL). the V/L surfaces of 48 molars were induced to an AWSL and divided in 6 groups, according to the conditioning protocols and application technique: 15% hydrochloric acid (HA) + manual application of the IR; HA + 37% phosphoric acid (PA) + manual application of the IR; HA + ultrasonic application (U) of the IR; HA + sonic application (S) of the IR; PA+HA+S; and PA+HA+U. For the Penetration Depth (PD), the crowns were etched with HA for 120s. The IR Icon® (DMG) was applied according to the manufacturer`s instructions. The crowns were dye penetrated (0.1% red fluorophore rhodamine B isothiocyanate for 12h) and bleached with 30% hydrogen peroxide for 12 h. The discs were immersed in a 50% ethanol solution, containing 100 µM of sodium fluorescein. The PD (in µm) was measured using confocal laser scanning microscopy (20x). The bond strength (BS) was performed by michoshear test (0.5 mm/min). Data were submitted to 2-way ANOVA and Tukey (α=0.05). For BS, the interaction was not significant (p>0.05). For PD, the main factors were significant (application - p<0.001; conditioning technique - p=0.003). The ultrasonic application showed the highest PD values. PA+HA presented higher results than HA. The sonic/ultrasonic applications and the use of phosphoric acid prior to hydrochloric acid improved PD of the infiltrant resin. Conditioning protocols or application techniques did not influence BS values.

Effect of conditioning protocols and ultrasonic application of an infiltrant resin in white spot lesions

Abstract The aim of this study was to evaluate different conditioning protocols and sonic/ultrasonic application of an infiltrant resin (IR) in artificial white spot lesions (AWSL). the V/L surfaces of 48 molars were induced to an AWSL and divided in 6 groups, according to the conditioning protocols and application technique: 15% hydrochloric acid (HA) + manual application of the IR; HA + 37% phosphoric acid (PA) + manual application of the IR; HA + ultrasonic application (U) of the IR; HA + sonic application (S) of the IR; PA+HA+S; and PA+HA+U. For the Penetration Depth (PD), the crowns were etched with HA for 120s. The IR Icon® (DMG) was applied according to the manufacturer`s instructions. The crowns were dye penetrated (0.1% red fluorophore rhodamine B isothiocyanate for 12h) and bleached with 30% hydrogen peroxide for 12 h. The discs were immersed in a 50% ethanol solution, containing 100 µM of sodium fluorescein. The PD (in µm) was measured using confocal laser scanning microscopy (20x). The bond strength (BS) was performed by michoshear test (0.5 mm/min). Data were submitted to 2-way ANOVA and Tukey (α=0.05). For BS, the interaction was not significant (p>0.05). For PD, the main factors were significant (application - p<0.001; conditioning technique - p=0.003). The ultrasonic application showed the highest PD values. PA+HA presented higher results than HA. The sonic/ultrasonic applications and the use of phosphoric acid prior to hydrochloric acid improved PD of the infiltrant resin. Conditioning protocols or application techniques did not influence BS values.

Resumo: O objetivo deste estudo foi avaliar diferentes protocolos de condicionamento e a aplicação sônica/ultrassônica de uma resina infiltrante (RI) em lesões de mancha branca produzidas artificialmente (LMBA). As superfícies vestibulares/linguais de 48 molares foram induzidas à formação de LMBA e divididas em 6 grupos, de acordo com os protocolos de condicionamento e técnica de aplicação da resina infiltrante: ácido hidroclorídrico 15% (AH) + aplicação manual da RI; AH + ácido fosfórico 37% (AF) + aplicação manual da RI; AH + aplicação ultrassônica da RI (U); AH + aplicação sônica da RI (S); AF + AH + S; e AF + AH + U. para o grau de penetração (GP), condicionou-se as coroas com AH por 120 s. A RI Icon® (DMG) foi aplicada de acordo com as instruções do fabricante. As coroas foram coradas (rodamina B 0,1% por 12 h) e clareadas com peróxido de hidrogênio 30% por 12 h. os discos foram imersos em solução de etanol 50%, contendo 100 µM de fluoresceína sódica. O GP (em µm) foi mensurado por meio de microscopia confocal a laser (20´). A resistência de união (RU) foi calculada pelo teste de microcisalhamento (0,5 mm/min). Os dados foram submetidos ao teste ANOVA 2 fatores e Tukey (α=0,05). Para RU, a interação não foi significante (p>0,05). Para GP, os fatores principais foram significantes (técnica de aplicação - p<0,001; protocolos de condicionamento - p=0,003). A aplicação U mostrou os maiores valores de GP. AF+AH demonstrou resultados superiores ao grupo AH. As aplicações sônica/ultrassônica e o uso do ácido fosfórico antes do ácido hidroclorídrico aumentaram o GP da resina infiltrante. Os protocolos de condicionamento ou as técnicas de aplicação não influenciaram os valores de RU.

Deformation behaviour of aged coronal dentin.

OBJECTIVE: This study aimed to identify the changes in the time-dependent deformation response of coronal dentin with ageing and its relationship with changes in chemical composition. BACKGROUND: The structural behaviour of dentin with ageing is affected by changes in the density and diameter of its dentinal tubules (ie porosity), as well as changes in chemical composition throughout the tooth. However, little is known about the time-dependent deformation behaviour of aged dentin and the importance of its hierarchical structure and variations in chemical composition. MATERIALS AND METHODS: The spherical indentation response of aged coronal dentin was analysed in the outer, middle and inner regions, and its time-dependent deformation response was modelled in terms of its microstructure and chemical composition using a model recently proposed for young dentin. RESULTS: The viscous deformation behaviour of aged dentin followed a power-law response with a decrease in the stress exponent when compared to young dentin. These results can be explained by cross-linking of the collagen present in the tissue. CONCLUSION: A decrease in the deformation ability of aged dentin was found. This behaviour could be a result of a dissolution process and reprecipitation of the minerals present in intertubular dentin into the dentinal tubules.

The PDK1-Rsk Signaling Pathway Controls Langerhans Cell Proliferation and Patterning.

Langerhans cells (LC), the dendritic cells of the epidermis, are distributed in a distinctive regularly spaced array. In the mouse, the LC array is established in the first few days of life from proliferating local precursors, but the regulating signaling pathways are not fully understood. We found that mice lacking the kinase phosphoinositide-dependent kinase 1 selectively lack LC. Deletion of the phosphoinositide-dependent kinase 1 target kinases, ribosomal S6 kinase 1 (Rsk1) and Rsk2, produced a striking perturbation in the LC network: LC density was reduced 2-fold, but LC size was increased by the same magnitude. Reduced LC numbers in Rsk1/2(-/-) mice was not due to accelerated emigration from the skin but rather to reduced proliferation at least in adults. Rsk1/2 were required for normal LC patterning in neonates, but not when LC were ablated in adults and replaced by bone marrow-derived cells. Increased LC size was an intrinsic response to reduced LC numbers, reversible on LC emigration, and could be observed in wild type epidermis where LC size also correlated inversely with LC density. Our results identify a key signaling pathway needed to establish a normal LC network and suggest that LC might maintain epidermal surveillance by increasing their "footprint" when their numbers are limited.

Structural and functional basis for p38-MK2-activated Rsk signaling in toll-like receptor-stimulated dendritic cells.

Rsk kinases play important roles in several cellular processes such as proliferation, metabolism, and migration. Until recently, Rsk activation was thought to be exclusively initiated by Erk1/2, but in dendritic cells (DC) Rsk is also activated by p38 mitogen-activated protein (MAP) kinase via its downstream substrates, MK2/3. How and why this noncanonical configuration of the MAP kinase pathway is adopted by these key immune cells are not known. We demonstrate that the Erk1/2-activated C-terminal kinase domain of Rsk is dispensable for p38-MK2/3 activation and show that compared with fibroblasts, a greater fraction of p38 and MK2/3 is located in the cytosol of DC prior to stimulation, suggesting a partial explanation for the operation of the noncanonical pathway of Rsk activation in these cells. p38/MK2/3-activated Rsk phosphorylated downstream targets and is physiologically important because in plasmacytoid DC (pDC) stimulated with Toll-like receptor 7 (TLR7) agonists, Erk1/2 activation is very weak relative to p38. As a result, Rsk activation is entirely p38 dependent. We show that this unusual configuration of MAP kinase signaling contributes substantially to production of type I interferons, a hallmark of pDC activation.

A crucial role for ß2 integrins in podosome formation, dynamics and Toll-like-receptor-signaled disassembly in dendritic cells.

The dynamic properties of podosomes, their ability to degrade the underlying matrix and their modulation by Toll-like receptor (TLR) signaling in dendritic cells (DCs) suggests they have an important role in migration. Integrins are thought to participate in formation and dynamics of podosomes but the multiplicity of integrins in podosomes has made this difficult to assess. We report that murine DCs that lack ß2 integrins fail to form podosomes. Re-expression of ß2 integrins restored podosomes but not when the membrane proximal or distal NPxF motifs, or when an intervening triplet of threonine residues were mutated. We show that ß2 integrins are remarkably long-lived in podosome clusters and form a persistent framework that hosts multiple actin-core-formation events at the same or adjacent sites. When ß2 integrin amino acid residues 745 or 756 were mutated from Ser to Ala, podosomes became resistant to dissolution mediated through TLR signaling. TLR signaling did not detectably modulate phosphorylation at these sites but mutation of either residue to phospho-mimetic Asp increased ß2 integrin turnover in podosomes, indicating that phosphorylation at one or both sites establishes permissive conditions for TLR-signaled podosome disassembly.