Detection of Microorganisms in Clinical Sonicated Orthopedic Devices Using Conventional Culture and qPCR / Detecção de microrganismos em dispositivos ortopédicos sonicados clínicos usando cultura convencional e qPCR

Abstract Objective To evaluate the sensitivity and specificity of the quantitative real-time polymerase chain reaction (qPCR) for 16S rDNA gene screening using sonicated fluid from orthopedic implants. Methods A retrospective study was conducted on 73 sonicated fluids obtained from patients with infection associated with orthopedic implants. The samples were subjected to conventional culture and molecular testing using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and qPCR for 16S rDNA. The cycle threshold values were used to define a cut-off of the qPCR of the 16S rDNA for negative and positive cultures. Results No statistical differences were observed between the positive and negative culture groups based on the time from the first surgery to infection (p= 0.958), age (p =0.269), or general comorbidities. Nevertheless, a statistical difference was found between the mean duration of antibiotic use before device removal (3.41 versus 0.94; p =0.016). Bacterial DNA was identified in every sample from the sonicated fluids. The median cycle thresholds of the positive and negative cultures were of 25.6 and 27.3 respectively (p< 0.001). As a diagnostic tool, a cycle threshold cut-off of 26.89 demonstrated an area under the curve of the receiver operating characteristic of 0.877 (p≤ 0.001). Conclusion The presence of antimicrobial agents for more than 72 hours decreased culture positivity, but did not influence the qPCR results. Despite this, amplification of the 16S rDNA may overestimate infection diagnosis.

Resumo Objetivo Avaliar a sensibilidade e a especificidade da reação em cadeia de polimerase em tempo real quantitativa (quantitative real-time polymerase chain reaction, qPCR, em inglês) para a triagem do gene rDNA 16S, com a utilização do fluido sonicado de implantes ortopédicos. Métodos Um estudo retrospectivo foi realizado em 73 fluidos sonicados obtidos de pacientes com infecção associada aos implantes ortopédicos. As amostras foram submetidas a cultura convencional e a teste molecular utilizando ionização e dessorção a laser assistida por matriz com espectrometria de massa por tempo de voo (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS, em inglês) e qPCR para o gene rDNA 16S. Os valores limiares do ciclo foram usados para definir um ponto de corte para a qPCR do gene rDNA 16S para culturas negativas e positivas. Resultados Não foram observadas diferenças estatísticas entre os grupos de cultura positiva e negativa com base no tempo desde a primeira cirurgia até a infecção (p= 0,958), na idade (p= 0,269), ou nas comorbidades em geral. No entanto, uma diferença estatística foi encontrada entre a duração média do uso de antibióticos antes da remoção do dispositivo (3,41 versus 0,94; p= 0,016). O DNA bacteriano foi identificado em todas as amostras dos fluidos sonicados. Os limiares do ciclo médio de culturas positivas e negativas foram de 25,6 e 27,3, respectivamente (p< 0,001). Como uma ferramenta de diagnóstico, um corte do limite do ciclo de 26,89 demonstrou uma área sob a curva da característica de operação do receptor de 0,877 (p ≤ 0,001). Conclusão A presença de agentes antimicrobianos por mais de 72 horas diminuiu a positividade da cultura, mas não influenciou os resultados da qPCR. Apesar disso, a amplificação do rDNA 16S pode sobrestimar o diagnóstico de infecção.

Evaluation of efficacy of new lens nucleus fragmenting forceps in cataract surgery / Avaliação da eficácia de uma nova pinça fragmentadora de núcleo do cristalino para cirurgia de catarata

ABSTRACT Objective: To evaluate the efficacy of low-cost nucleus fragmenting forceps designed to reduce the use of ultrasound during phacoemulsification. Methods: A total of 60 pig eyes enucleated 10 hours before were placed in the microwave oven, at maximum power for 10 seconds, to form cataracts with hardness comparable to a grade IV nucleus in the Lens Opacities Classification System III. Cataract extraction was performed using the Centurion® phacoemulsifier (Alcon Laboratories, Geneve, Switzerland) and Leica M620 microscope. All eyes were submitted to the pre-chop technique dividing the nucleus into four parts. After the pre-chop, the quadrants in 30 eyes were phacoemulsified with the torsional mode and were fragmented in the remaining 30 eyes after the pre-chop was with the fragmentation forceps before torsional mode phacoemulsification. The device was calibrated for all eyes by applying the following parameters: 40% linear torsional phacoemulsification; intraocular pressure of 65 mmHg; the linear vacuum of 600 mmHg; aspiration flow of 40 ccs/minute. After each procedure, the following was recorded: cumulative dissipated energy; equivalent average torsional amplitude; equivalent average ultrasonic power; estimated aspirated fluid; ultrasound total time; and total aspiration time. Statistical analysis was performed using the Kruskal-Wallis test and the IBM Statistical Package for Social Sciences. The p-value <0.05 was considered statistically significant. Results: There was a statistically significant reduction favoring the use of the nucleus fragmenting forceps in all parameters, except for the average torsional amplitude. Conclusion: The use of the nucleus fragmenting forceps contributed to improving the efficacy of torsional phacoemulsification in enucleated pig eyes.

RESUMO Objetivo: Avaliar a eficácia de uma pinça fragmentadora de núcleo, de baixo custo, desenvolvida para reduzir o uso de ultrassom durante a emulsificação do núcleo. Métodos: Sessenta olhos de porco com 10 horas de enucleação foram colocados no forno de microondas, em potência máxima por 10 segundos, para a formação de catarata com dureza comparável à de um núcleo grau IV na Lens Opacities Classification System III. A extração da catarata foi realizada com o facoemulsificador Centurion® (Alcon Laboratories, Genebra, Suíça) e microscópio Leica M620. Todos os olhos foram submetidos a técnica de pre-chop, dividindo o núcleo em quatro partes. Em 30 olhos, após o pre-chop, foi feita a facoemulsificação dos quadrantes com o modo torsional e, nos outros 30 olhos, após o pre-chop, cada quadrante foi fragmentado com a pinça antes da facoemulsificação com o modo torsional. O aparelho foi calibrado para todos os olhos com os seguintes parâmetros: faco torsional linear 40%; pressão intraocular 65 mmHg; vácuo linear 600mmHg e fluxo de aspiração 40cc/minuto. Após cada procedimento, verificaram-se energia dissipada acumulada; média da amplitude do faco torsional; média equivalente do poder ultrassônico; líquido aspirado estimado; tempo total de ultrassom e tempo total de aspiração. A análise estatística foi realizada utilizando o teste de Kruskal-Wallis com o IBM Statistical Package for Social Sciences. Valor de p<0,05 foi considerado estatisticamente significante. Resultados: Houve redução estatisticamente significante em favor do uso da pinça fragmentadora de núcleo em todos os parâmetros, menos na média de amplitude do faco torsional. Conclusão: O uso da pinça fragmentadora de núcleo contribuiu para melhorar a eficácia do faco torsional em olhos de porco enucleados.

Efeito do BioGran® funcionalizado ao raloxifeno através do processamento ultrassônico no reparo ósseo em defeitos críticos de calvárias de ratos / Effect of BioGran® functionalized to Raloxifene through ultrasonic processing in bone repair in critical defects in rat calvaria

Este estudo teve como objetivo analisar o processo de formação óssea, bem como a microarquitetura óssea promovido pela associação entre o BioGran® e diferentes concentrações de raloxifeno através do método da sonoquímica em defeitos críticos realizados em calvária de rato. Em um total de doze ratos machos, um defeito crítico de 5 mm de diâmetro foi feito e preenchido com BioGran® 100% (Bg), BioGran® 90% associado à Raloxifeno 10%, e BioGran® 80% associado à Raloxifeno 20%. Aos 14 e 24 dias pós-operatório, foram aplicados os fluorocromos calceína e alizarina, de modo respectivo. A eutanásia ocorreu aos 30 dias após a realização do procedimento cirúrgico para enxertia dos biomateriais. Análise de microtomografia computadorizada (micro-CT) através dos parâmetros de superfiície de intersecção (i.S), fração de volume ósseo (BV/TV) e densidade de conectividade (Conn.Dn) e por microscopia confocal a laser por meio da dinâmica óssea, superfície de mineralização ativa e a taxa de aposição mineral (MAR). Os dados foram analisados através de análise estatística utilizando o teste de normalidade Shapiro-Wilk e o pós teste de Turkey (p<0.05). Para os parâmetros de micro-CT avaliados os menores valores foram encontrados no grupo BG+RL10% (p<0,05), valores similares foram encontrados entre os grupos BG e BG+RL20%. A microscopia confocal evidenciou melhor mineralização óssea e maior taxa de aposição mineral (MAR) para o Grupo BG+RL20% (p<0,05%). Conclusão: A concentração de Raloxifeno a 20% combinado ao BioGran® pelo método da sonoquímica parece ter acelerado o reparo ósseo(AU)

This study aimed to analyze the bone formation process, as well as the bone microarchitecture promoted by the association between BioGran® and different concentrations of raloxifene through the sonochemistry method in critical defects performed in rat calvaria. In a total of twelve male rats, a critical defect of 5 mm in diameter was made and filled with BioGran® 100% (Bg), BioGran® 90% associated with Raloxifene 10%, and BioGran® 80% associated with Raloxifene 20%. At 14 and 24 days postoperatively, the fluorochromes calcein and alizarin were applied, respectively. Euthanasia occurred 30 days after the surgical procedure for grafting biomaterials. Analysis of computed microtomography (microCT) through the parameters of intersection surface (iS), bone volume fraction (BV / TV) and connectivity density (Conn.Dn) and by confocal microscopy through bone dynamics, surface of active mineralization and the mineral apposition rate (MAR). Data were analyzed through statistical using the Shapiro-Wilk normality test and the Turkey post-test (p <0.05). For the me microCT parameters evaluated, the lowest values were found in the BG + RL10% group (p<0.05), similar values were found between the BG and BG + RL20% groups. Confocal microscopy showed better bone mineralization and higher mineral apposition rate (MAR) for the BG + RL20% Group (p < 0.05%). Conclusion: The concentration of Raloxifene at 20% combined with BioGran® by the sonochemistry method seems to have accelerated bone repair(AU)

Infection of expanded polytetrafluoroethylene and Dacron-coated stents with Staphylococcus epidermidis: an experimental study in pigs / Infecção de stents revestidos com politetrafluoretileno expandido e Dacron com Staphylococcus epidermidis: estudo experimental em porcos

Abstract Background Diagnosis of the etiologic agent of endoprosthesis infections is essential to enable treatment, since these infections constitute important complications of endovascular procedures. Sonication of explanted tissue and materials is a technique that can be used to facilitate detection of biofilm-producing bacteria. Objectives To evaluate infection of pigs' aortas after implantation of nitinol stents coated with polytetrafluoroethylene (ePTFE) or Dacron, previously infected with biofilm-producing Staphylococcus epidermidis. Intimal thickening and the inflammatory response in the aortic wall were also evaluated. Methods 11 ePTFE-coated nitinol stents and 10 Dacron stents infected with S. epidermidis strains were implanted in the infrarenal aorta of 21 8-week-old pigs. After 2 weeks, the aorta containing the stents was removed. A vortex mixer and ultrasound were used to homogenize the samples and remove the biofilm. Subsequently, the number of colony-forming units was counted. Results There were no significant differences between the two groups in terms of the number of colony-forming units or of inflammation in the arterial wall. With the exception of one specimen from the Dacron group, all aortic stent cultures were positive for S. epidermidis. Conclusions There were no significant differences in the inflammatory response or infection rate between ePTFE and Dacron-coated stents actively infected with biofilm-producing S. epidermidis. Intimal thickening and the inflammatory response to infection of endoprostheses were similar. These results suggest that the two most widely used stent lining materials have a similar infection rate.

Resumo Contexto O diagnóstico do agente etiológico é essencial para o tratamento das infecções de endoprótese, pois representam uma importante complicação do tratamento endovascular. A sonificação dos tecidos pode ser uma técnica usada para auxiliar na detecção de bactérias produtoras de biofilme. Objetivos Avaliar a infecção da aorta dos porcos após o implante de stents de nitinol revestidos com politetrafluoretileno (ePTFE) ou Dacron, infectados com Staphylococcus epidermidis, produtor de biofilme. O espessamento intimal e a resposta inflamatória na parede aórtica também foram avaliados. Métodos Onze stents de nitinol revestidos com ePTFE e 10 stents de Dacron infectados com cepas de S. epidermidis foram implantados na aorta infrarrenal de 21 porcos com 8 semanas de idade. Após duas semanas, a aorta contendo os stents foi removida. Um misturador de vórtice e ultrassom foram utilizados para homogeneizar as amostras e remover o biofilme. Posteriormente, o número de unidades formadoras de colônias foi contado. Resultados Não houve diferenças significativas no número de unidades formadoras de colônias ou inflamação na parede arterial entre os dois grupos. Todas as culturas de stent aórtico foram positivas para S. epidermidis, exceto uma no grupo Dacron. Conclusões Não houve diferenças significativas na resposta inflamatória ou na taxa de infecção entre os stents revestidos de ePTFE e Dacron, infectados ativamente pelo S. epidermidis produtor de biofilme. O espessamento intimal e a resposta inflamatória à infecção das endopróteses foram semelhantes. Esses resultados sugerem que os dois materiais de revestimento de stent mais amplamente utilizados têm uma taxa de infecção semelhante.

Influence of sonic vibration and cement type on the bond strength of fiberglass posts after aging / Influencia de la vibración sónica y el tipo de cemento en la fuerza de unión de los postes de fibra de vidrio después del envejecimiento

Introduction: This study evaluated the influence of sonic vibration and thermocycling on the bond strength of fiberglass posts using conventional dual-cured and self-adhesive resin cements. Material and Methods: Ninety-six single-rooted endodontically treated teeth were divided randomly into eight groups according to the cement used (dual-cured or self-adhesive resin cement), use of sonic vibration during post accommodation, and aging conditions (24h in distilled water or 5,000 thermal cycles). The fiberglass posts (White Post DC1, FGM) were cleaned with alcohol, treated with silane and cemented with dual-cured (Allcem Core, FGM) or with self-adhesive resin cement (seT, SDI). For groups in which sonic vibration was used, the posts were accommodated, and sonic vibration was applied for 10 s using a special tip placed on top of the post (Sonic Smart Device, FGM). Pull-out tests were performed after storage in distilled water for 24h at 37°C or after thermocycling (5000 cycles, at 5°C and 55°C). The results were evaluated using three-way ANOVA and Tukey's test (α=0.05). Results: Significant differences were not observed between the bond strengths of dual-cured (87.93±41.81 N) and self-adhesive cement (82.53±41.43 N). Bond strength for the sonic vibration groups (100.36±42.35 N) was significantly higher than for groups without sonic vibration (70.13±34.90 N). There were significant differences between specimens subjected to thermocycling (98.33±39.42 N) and those stored for 24h in distilled water (72.16±39.67 N). Conclusion: It can be concluded that both sonic vibration and thermocycling significantly improved bond strength of fiberglass posts with the two evaluated resin cements.

Introducción: Este estudio evaluó la influencia de la vibración sónica y el termociclado en la fuerza de unión de los postes de fibra de vidrio utilizando cementos de resina autoadhesivos y de curado dual convencionales Material y Métodos: Se dividieron aleatoriamente noventa y seis dientes tratados endodónticamente de raíz única en ocho grupos según el cemento utilizado (cemento de resina de curado dual o autoadhesivo), el uso de vibración sónica durante la acomodación posterior y las condiciones de envejecimiento (24 horas agua destilada o 5.000 ciclos térmicos). Los postes de fibra de vidrio (White Post DC1, FGM) se limpiaron con alcohol, se trataron con silano y se cementaron con cemento de curado dual (Allcem Core, FGM) o con cemento de resina autoadhesivo (seT, SDI). Para los grupos en los que se utilizó vibración sónica, se acomodaron los postes y se aplicó la vibración sónica durante 10 s utilizando una punta especial colocada en la parte superior del poste (Sonic Smart Device, FGM). Las pruebas de extracción se realizaron después del almacenamiento en agua destilada durante 24 horas a 37ºC o después del termociclado (5000 ciclos, a 5ºC y 55ºC). Los resultados se evaluaron mediante ANOVA de tres vías y la prueba de Tukey (α = 0.05). Resultados: No se observaron diferencias significativas entre las resistencias de adhesión del cemento de curado dual (87,93 ± 41,81 N) y el cemento autoadhesivo (82,53 ± 41,43 N). La fuerza de unión para los grupos de vibración sónica (100,36 ± 42,35 N) fue significativamente mayor que para los grupos sin vibración sónica (70,13 ± 34,90 N). Hubo diferencias significativas entre los especímenes sometidos a termociclado (98,33 ± 39,42 N) y los almacenados durante 24 h en agua destilada (72,16 ± 39,67 N). Conclusión: Se puede concluir que tanto la vibración sónica como el termociclado mejoraron significativamente la fuerza de unión de los postes de fibra de vidrio con los dos cementos de resina evaluados.

Improvement of Mycobacterium tuberculosis detection in sputum using DNA extracted by sonication

Abstract Tuberculosis (TB) is one of the infectious diseases with high mortality in the world. DNA amplification techniques have been used to overcome barriers to the diagnosis of this disease. However, the success of these methodologies is highly dependent on the DNA obtained from the sample. This study was carried out to verify whether the DNA extracted by sonication (in house method) could yield suitable DNA for amplification by real-time PCR (qPCR). Sixty sputum samples were submitted to DNA extraction using sonication compared to a commercial method (Detect-TB kit, Labtest/MG-Brazil). All DNA samples were amplified by qPCR for IS6110 region (IS6110-qPCR/SYBR Green assay). Out of 60 samples, 40 were positive for TB; of these, all had positive results when extracted by sonication (100%) and 80% when extracted by the commercial method. The limit of detection (LOD) of Mycobacterium tuberculosis (H37Rv strain) by qPCR was 14CFU/mL when the DNA was extracted by sonication, compared to countless colonies when extracted by commercial kit. In conclusion, the sonication protocol (without purification step) proved to be a simple, fast, and suitable method for obtaining DNA for use in qPCR from sputum samples.

Crecimiento y metabolismo de Fischerella TB22 en medio de cultivo BG11 / Growth and metabolism of Fischerella TB22 in BG110 culture medium

Los microorganismos fijadores de nitrógeno de vida libre, abarcan una gama morfológica que va desde los organismos unicelulares como las bacterias y algunas cianobacterias, hasta multicelulares, filamentosas, por ello es importante conocer cómo se comportan y se puede saber haciendo una curva de crecimiento microbiano. Para este estudio se prepararon 4 fotobioreactores de columna burbujeada con inoculo de Fischerella TB22, se pusieron en aireación constante con 12 horas luz y 12 horas obscuridad durante 40 días con diferentes tratamientos de ajuste de volumen del medio de cultivo y ajuste del pH. El objetivo de este trabajo fue evaluar el crecimiento en biomasa por peso seco, densidad óptica, pH y amonio de Fischerella sp. en medio de cultivo BG110 durante 12 días. Las variables que se midieron de la curva de crecimiento de las cianobacterias, siguieron el patrón de una curva típica de crecimiento microbiano. (AU)

Free-living nitrogen-fixing microorganisms cover a morphological range that goes from unicellular organisms such as bacteria and some cyanobacteria, to multicellular, filamentous, therefore it is important to know how they behave and can be known by makinga microbial growth curve. For this study, 4 bubbled column photobioreactors with Fischerella TB22inoculum were prepared, they were placed in constant aeration with 12 hours of light and 12 hours of darkness for 40 days with different treatments of volumeadjustment of the culture medium and pH adjustment. The objective of this work was to evaluate the biomass growth by dry weight, optical density, pH, and ammonia of Fischerella sp. in the BG110 culture medium for 12 days. The variables that were measured from the growth curve of cyanobacteria followed the pattern of a typical microbial growth curve. (AU)

Micro and nanocrystalline cellulose based oral dispersible film; preparation and evaluation of in vitro/in vivo rapid release studies for donepezil

Oral fast-dispersible film was prepared by utlizing donepezil hydrochloride (drug) and various cellulose derivatives such as hydroxypropyl methyl cellulose (hypermellose) (HPMC), microcrystalline cellulose (MCC) and nanocrystalline cellulose (NCC) to treat Alzheimer's disease. NCC was synthesized by ultra-sonication method using MCC and this was converted to thinfilm formulation (NCC-F) using solvent casting technique. The interaction between the polymer and the drug was investigated by spectral analysis such as UV, FTIR, and 1H- NMR. FTIR confirmed that the compatibility of drug and polymer in ODF formulation. NCC-F has shown an average surface roughness of 77.04 nm from AFM and the average particle size of 300 nm from SEM analysis. Nano sized particle of NCC-F leads faster in vitro dissolution rate (94.53%) when compared with MCC-F and F3 formulation. Animal model (in vivo) studies of NCC-F formulation has reached peak plasma concentration (Cmax) up to 19.018 ng/mL in the span of (tmax) 4 h with greater relative bioavailability of 143.1%. These results suggested that high surface roughness with nanosized NCC-F formulation attained extended drug availability up to (t1/2) 70 h.

Factors Related to Successful Energy Transmission of Focused Ultrasound through a Skull: A Study in Human Cadavers and Its Comparison with Clinical Experiences

OBJECTIVE: Although magnetic resonance guided focused ultrasound (MRgFUS) has been used as minimally invasive and effective neurosurgical treatment, it exhibits some limitations, mainly related to acoustic properties of the skull barrier. This study was undertaken to identify skull characteristics that contribute to optimal ultrasonic energy transmission for MRgFUS procedures.METHODS: For ex vivo skull experiments, various acoustic fields were measured under different conditions, using five non-embalmed cadaver skulls. For clinical skull analyses, brain computed tomography data of 46 patients who underwent MRgFUS ablations (18 unilateral thalamotomy, nine unilateral pallidotomy, and 19 bilateral capsulotomy) were retrospectively reviewed. Patients' skull factors and sonication parameters were comparatively analyzed with respect to the cadaveric skulls.RESULTS: Skull experiments identified three important factors related skull penetration of ultrasound, including skull density ratio (SDR), skull volume, and incidence angle of the acoustic rays against the skull surface. In clinical results, SDR and skull volume correlated with maximal temperature (Tmax) and energy requirement to achieve Tmax (p<0.05). In addition, considering the incidence angle determined by brain target location, less energy was required to reach Tmax in the central, rather than lateral targets particularly when compared between thalamotomy and capsulotomy (p<0.05).CONCLUSION: This study reconfirmed previously identified skull factors, including SDR and skull volume, for successful MRgFUS; it identified an additional factor, incidence angle of acoustic rays against the skull surface. To guarantee successful transcranial MRgFUS treatment without suffering these various skull issues, further technical improvements are required.

Magnetic Resonance-Guided Focused Ultrasound : Current Status and Future Perspectives in Thermal Ablation and Blood-Brain Barrier Opening

Magnetic resonance-guided focused ultrasound (MRgFUS) is an emerging new technology with considerable potential to treat various neurological diseases. With refinement of ultrasound transducer technology and integration with magnetic resonance imaging guidance, transcranial sonication of precise cerebral targets has become a therapeutic option. Intensity is a key determinant of ultrasound effects. High-intensity focused ultrasound can produce targeted lesions via thermal ablation of tissue. MRgFUS-mediated stereotactic ablation is non-invasive, incision-free, and confers immediate therapeutic effects. Since the US Food and Drug Administration approval of MRgFUS in 2016 for unilateral thalamotomy in medication-refractory essential tremor, studies on novel indications such as Parkinson's disease, psychiatric disease, and brain tumors are underway. MRgFUS is also used in the context of blood-brain barrier (BBB) opening at low intensities, in combination with intravenously-administered microbubbles. Preclinical studies show that MRgFUS-mediated BBB opening safely enhances the delivery of targeted chemotherapeutic agents to the brain and improves tumor control as well as survival. In addition, BBB opening has been shown to activate the innate immune system in animal models of Alzheimer’s disease. Amyloid plaque clearance and promotion of neurogenesis in these studies suggest that MRgFUS-mediated BBB opening may be a new paradigm for neurodegenerative disease treatment in the future. Here, we review the current status of preclinical and clinical trials of MRgFUS-mediated thermal ablation and BBB opening, described their mechanisms of action, and discuss future prospects.

Efficacy of sonic and ultrasonic irrigation devices in the removal of debris from canal irregularities in artificial root canals

Abstract Objective To evaluate the efficacy of different sonic and ultrasonic devices in the elimination of debris from canal irregularities in artificial root canals. Materials and Methods A resin model of a transparent radicular canal filled with dentin debris was used. Five groups were tested, namely: Group 1 - ultrasonic insert 15.02; Group 2 - ultrasonic insert 25/25 IRRI K; Group 3 - ultrasonic insert 25/25 IRRI S; Group 4 - sonic insert 20/28 Eddy on a vibrating sonic air-scaler handpiece; Group 5 - 20.02 K-file inserted on a Safety M4 handpiece. Two different irrigants (5% sodium hypochlorite and 17% EDTA) and 3 different times of activation (20, 40, and 60 seconds) were tested. Means and standard deviations were calculated and statistically analyzed with the Kruskal-Wallis and Wilcoxon tests (p<0.05). Results No statistically significant differences were found between the two irrigants used. Group 4 removed more debris than the other groups (p<0.05). Groups 1, 2, and 3 removed more debris than group 5 (p<0.05). A statistically significant difference (p<0.05) was found for the time of activation in all groups and at all canal levels, except between 40 and 60 seconds in group 4 at coronal and middle third level (p>0.05). Conclusions No significant differences were found between 5% sodium hypochlorite and 17% EDTA. When the time of activation rises, the dentin debris removal increases in all groups. Both sonic and ultrasonic activation demonstrate high capacity for dentin debris removal.

Factors Related to Successful Energy Transmission of Focused Ultrasound through a Skull: A Study in Human Cadavers and Its Comparison with Clinical Experiences

OBJECTIVE: Although magnetic resonance guided focused ultrasound (MRgFUS) has been used as minimally invasive and effective neurosurgical treatment, it exhibits some limitations, mainly related to acoustic properties of the skull barrier. This study was undertaken to identify skull characteristics that contribute to optimal ultrasonic energy transmission for MRgFUS procedures. METHODS: For ex vivo skull experiments, various acoustic fields were measured under different conditions, using five non-embalmed cadaver skulls. For clinical skull analyses, brain computed tomography data of 46 patients who underwent MRgFUS ablations (18 unilateral thalamotomy, nine unilateral pallidotomy, and 19 bilateral capsulotomy) were retrospectively reviewed. Patients' skull factors and sonication parameters were comparatively analyzed with respect to the cadaveric skulls. RESULTS: Skull experiments identified three important factors related skull penetration of ultrasound, including skull density ratio (SDR), skull volume, and incidence angle of the acoustic rays against the skull surface. In clinical results, SDR and skull volume correlated with maximal temperature (Tmax) and energy requirement to achieve Tmax (p<0.05). In addition, considering the incidence angle determined by brain target location, less energy was required to reach Tmax in the central, rather than lateral targets particularly when compared between thalamotomy and capsulotomy (p<0.05). CONCLUSION: This study reconfirmed previously identified skull factors, including SDR and skull volume, for successful MRgFUS; it identified an additional factor, incidence angle of acoustic rays against the skull surface. To guarantee successful transcranial MRgFUS treatment without suffering these various skull issues, further technical improvements are required.

Magnetic Resonance-Guided Focused Ultrasound : Current Status and Future Perspectives in Thermal Ablation and Blood-Brain Barrier Opening

Magnetic resonance-guided focused ultrasound (MRgFUS) is an emerging new technology with considerable potential to treat various neurological diseases. With refinement of ultrasound transducer technology and integration with magnetic resonance imaging guidance, transcranial sonication of precise cerebral targets has become a therapeutic option. Intensity is a key determinant of ultrasound effects. High-intensity focused ultrasound can produce targeted lesions via thermal ablation of tissue. MRgFUS-mediated stereotactic ablation is non-invasive, incision-free, and confers immediate therapeutic effects. Since the US Food and Drug Administration approval of MRgFUS in 2016 for unilateral thalamotomy in medication-refractory essential tremor, studies on novel indications such as Parkinson's disease, psychiatric disease, and brain tumors are underway. MRgFUS is also used in the context of blood-brain barrier (BBB) opening at low intensities, in combination with intravenously-administered microbubbles. Preclinical studies show that MRgFUS-mediated BBB opening safely enhances the delivery of targeted chemotherapeutic agents to the brain and improves tumor control as well as survival. In addition, BBB opening has been shown to activate the innate immune system in animal models of Alzheimer’s disease. Amyloid plaque clearance and promotion of neurogenesis in these studies suggest that MRgFUS-mediated BBB opening may be a new paradigm for neurodegenerative disease treatment in the future. Here, we review the current status of preclinical and clinical trials of MRgFUS-mediated thermal ablation and BBB opening, described their mechanisms of action, and discuss future prospects.

Skull Factors Affecting Outcomes of Magnetic Resonance-Guided Focused Ultrasound for Patients with Essential Tremor

PURPOSE: Magnetic resonance-guided focused ultrasound (MRgFUS) thalamotomy has become a standard treatment for medically intractable essential tremor (ET). Skull density ratio (SDR) and skull volume in patients with ET are currently considered useful indicators of the successful application of MRgFUS. We compared the clinical outcomes of MRgFUS thalamotomy with SDR above 0.4 and 0.45. We also described patterns of SDR and skull volume in Korean patients with ET who were eligible to be screened for MRgFUS. MATERIALS AND METHODS: In screening 318 ET patients, we evaluated patterns of skull density and skull volume according to age and sex. Fifty patients with ET were treated with MRgFUS. We investigated the effects of SDR and skull volume on treatment parameters and the outcomes of ET. RESULTS: The mean SDR of the 318 ET patients was 0.45±0.11, and that for skull volume was 315.74±40.95 cm³. The male patients had a higher SDR than female patients (p=0.047). Skull volume significantly decreased with aging. SDR and skull volume exhibited a linear negative relationship. Among therapeutic parameters, maximal temperature was positively related to SDR, while sonication number was not related to either SDR or skull volume. Tremor outcome was also not related to SDR or skull volume. CONCLUSION: SDR varied widely from 0.11 to 0.73, and men had a higher SDR. Therapeutic parameters and clinical outcomes were not affected by SDR or skull volume.

Effect of sonic versus ultrasonic activation on aqueous solution penetration in root canal dentin

The aim of this study was to observe the penetration of an aqueous solution into the root canal dentin under sonic activation and ultrasonic activation. Materials and Method: this study consisted of experimental in vitro research. In order to achieve a closed system, the apex of 45 single-rooted teeth was sealed with wax. The step-back technique was manually performed using a K50 apical master file and 3 groups were organized according to the protocol of the final irrigant activation: group I: non-activated chinese ink for 30 seconds, group II: chinese ink sonically activated with EndoActivator for 30 seconds, and group III: chinese ink ultrasonically activated with Varios 350 equipment for 30 seconds. Teeth were sectioned longitudinally, and the samples obtained were observed under a stereomicroscope at 1X magnification in order to be photographed and scanned to calculate the penetration area using the Image J software. The tinted radicular area was evaluated in relation to the total area of the root dentin. The tukey's post-hoc test and ANOVA were used for the statistical analysis (p<0.05). Results: group I and II obtained 9.13 percent and 9.42 percent penetration respectively, while in group III the highest degree of dye infiltration was achieved (13.9 percent), being statistically significant (p<0.001). Conclusions: ultrasonic activation produced a significantly higher penetration of the dye when compared to conventional activation and sonic activation.

The effects of proteins released from silk mat layers on macrophages / 대한악안면성형재건외과학회지

BACKGROUND: The objective of this study was to evaluate the changes in gene expression after incubation of cells with proteins released from different silk mat layers. METHODS: A silk cocoon from Bombyx mori was separated into four layers of equal thickness. The layers were numbered from 1 to 4 (from the inner to the outer layer). The proteins were released by sonication of a silk mat layer in normal saline. The concentration of proteins was determined by spectrophotometry. They were incubated with RAW264.7 cells, and changes in the expression of genes were evaluated by cDNA microarray analysis and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). RESULTS: Layer 1 and 4 groups had higher protein concentrations compared to those in layer 2 and 3 groups. The genes associated with inflammation and angiogenesis showed significantly higher expression in layer 1 and 4 groups. The results of qRT-PCR were in agreement with those of the cDNA microarray analysis. CONCLUSIONS: The silk mat from the middle portion of the silkworm cocoon yielded a lower protein release and caused an insignificant change in the expression of genes that are associated with inflammation and angiogenesis.

Safety of Temporary Use of Recycled Autoclaved Femoral Components in Infected Total Knee Arthroplasty: Confirming Sterility Using a Sonication Method

BACKGROUND: The purpose of this study was to evaluate the usefulness of sonication technique for microbiological diagnosis and the sterility of the recycled autoclaved femoral components from infected total knee arthroplasty (TKA) using a sonication method. METHODS: Nineteen femoral implants explanted from patients with infected TKA were sterilized with a standard autoclave method. Standard culture of the fluid before and after sonication of the sterilized implants was performed to detect pathogenic microorganisms. Additional experiments were performed to evaluate the sterility of the recycled implant by inducing artificial biofilm formation. Methicillin-resistant Staphylococcus aureus (MRSA) was inoculated into 10 implants and sterilization in a standard autoclave was performed, and then the fluid was cultured before and after sonication. RESULTS: Two of the 19 sterilized implants were positive for growth of bacteria after sonication, whereas no growth was detected in the cultured fluid from the sterilized implants before sonication. The bacteria were Staphylococcus species in all two cases. In one of 10 implants inoculated with MRSA, the culture was positive for growth of bacteria both before and after sonication. However, Staphylococcus epidermidis was cultured from both occasions and thus this implant was thought to be contaminated. CONCLUSIONS: We found sonication for identification of pathogens could be helpful, but this finding should be interpreted carefully because of the possibility of contamination. Sterilization of an infected femoral implant with an autoclave method could be a good method for using the temporary articulating antibiotic spacer in two-stage revision arthroplasty.

Technical Review and Perspectives of Transcranial Focused Ultrasound Brain Stimulation for Neurorehabilitation

Lack of a region-specific brain stimulation modality having both spatial specificity and depth penetration has limited clinicians to explore novel non-pharmacological treatment options in neurorehabilitation. Focused ultrasound (FUS) has shown excitatory and suppressive modulatory effects on neural tissues in both central and peripheral nervous systems by transcranially delivering low-intensity highly focused acoustic pressure waves to region-specific neural structures in a completely non-invasive fashion. This emerging technique, with exquisite spatial selectively and depth penetration, is considered as a new mode of brain stimulation that may significantly improve existing brain stimulation modalities. This review aims to provide the perspectives of FUS-mediated brain stimulation in neurorehabilitation, along with potential pitfalls and cautions that need to be taken into consideration. When combined with the intravascular introduction of microbubble-based ultrasound contrast agents, the technique adds therapeutic potentials in delivering drug/genes/cells across the blood-brain barrier, which may open new opportunities for neurorehabilitation. Efforts are being made to construct FUS devices appropriate for routine clinical use, to investigate its fundamental mechanisms, and to optimize the sonication parameters. Repeated administration of the technique for inducing neuroplasticity, including the assessment of long-term safety, is warranted to reveal its utility in neurorehabilitation.

Efficacy, Efficiency, and Safety of Magnetic Resonance-Guided High-Intensity Focused Ultrasound for Ablation of Uterine Fibroids: Comparison with Ultrasound-Guided Method

OBJECTIVE: The purpose of this study was to compare efficacy, sonication energy efficiency, treatment time and safety of magnetic resonance-guided high-intensity focused ultrasound (MRgHIFU) and those of ultrasound-guided high-intensity focused ultrasound (USgHIFU) for ablation of uterine fibroids. MATERIALS AND METHODS: This study included 43 patients with 44 symptomatic uterine fibroids treated with MRgHIFU and 51 patients with 68 symptomatic uterine fibroids treated with USgHIFU. After therapy, contrast-enhanced MRI was conducted and complete ablation was defined as 100% non-perfused volume (NPV) of fibroids. Patients with completely ablated fibroids were selected for the comparison of the treatment data and sonication parameters between MRgHIFU and USgHIFU treated groups. RESULTS: Thirteen completely ablated fibroids in 10 patients (23.3%, 10/43) were achieved with MRgHIFU and 28 completely ablated fibroids in 22 patients (43.1%, 22/51) were achieved with USgHIFU. In completely ablated fibroids, the energy-efficiency factor (EEF) was 5.1 ± 3.0 J/mm3 and 4.7 ± 2.5 J/mm3 in the MRgHIFU and USgHIFU, respectively (p = 0.165). There was a negative linear correlation between EEF and the NPV of fibroids for MRgHIFU (p = 0.016) and USgHIFU (p = 0.001). The mean treatment time was 174.5 ± 42.2 minutes and 114.4 ± 39.2 minutes in the MRgHIFU and USgHIFU procedures, respectively (p = 0.021). There were no severe adverse events and major complications after treatment. CONCLUSION: MRgHIFU and USgHIFU are safe and effective with the equivalent energy efficiency for complete ablation of fibroids. USgHIFU has shorter treatment time than MRgHIFU.

Synthesis of ultrasound contrast agents: characteristics and size distribution analysis (secondary publication)

PURPOSE: The purpose of this study was to establish a method for ultrasound (US) contrast agent synthesis and to evaluate the characteristics of the synthesized US contrast agent. METHODS: A US contrast agent, composed of liposome and sulfur hexafluoride (SF₆), was synthesized by dissolving 21 μmol 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine (DPPC, C₄₀H₈₀NO₈P), 9 μmol cholesterol, and 1.9 μmol of dihexadecylphosphate (DCP, [CH₃(CH₂)15O]₂P(O)OH) in chloroform. After evaporation in a warm water bath and drying for 12-24 hours, the contrast agent was synthesized using the sonication process by the addition of a buffer and SF₆ gas. The size distribution of the bubbles was analyzed using dynamic light scattering measurement methods. The degradation curve was evaluated by assessing the change in the number of contrast agent bubbles using light microscopy immediately, 12, 24, 36, 48, 60, 72, and 84 hours after synthesis. The echogenicity of the synthesized microbubbles was compared with commercially available microbubbles (SonoVue, Bracco). RESULTS: contrast agent was synthesized successfully using an evaporation-drying-sonication method. Most bubbles had a mean diameter of 154.2 nm and showed marked degradation 24 hours after synthesis. Although no statistically significant differences were observed between SonoVue and the synthesized contrast agent, a difference in echogenicity was observed between the synthesized contrast agent and saline (P<0.01). CONCLUSION: We successfully synthesized a US contrast agent using an evaporation-dryingsonication method. These results may help future research in the fields of anticancer drug delivery, gene delivery, targeted molecular imaging, and targeted therapy.