Fluorescencia con verde de indocianina intravesicular en colecistectomía laparoscópica / Fluorescence with intravesicular indocyanine green in laparoscopic cholecystectomy

La investigación trata de una serie de casos realizados de manera prospectiva y observacional, para establecer la eficacia del uso de fluorescencia con verde de indocianina intravesicular en 15 pacientes sometidos a colecistectomía laparoscópica en la Catedra de Clínica y Terapéutica Quirúrgica A, servicio de cirugía I del hospital universitario de Caracas entre junio y agosto de 2024. A todos los pacientes se les administró 2,5 mg de verde de indocianina, la ampolla de verde de indocianina Aldagor® de 25mg fue diluida en 10 ml de solución salina fisiológica, por vía intravesicular con una aguja espinal 25G utilizando torre laparoscópica OptoMedic® FloNavi 214k. La técnica se aplicó de forma estandarizada en todos los casos, antes de la disección del triángulo de Calot. Resultados: La Fluorescencia se logró en la totalidadde los casos, en el fondo y cuerpo vesicular posterior a la infiltración del verde de indocianina intravesicular la fluorescencia fue a los 2.4 minutos, el tiempo de fluorescencia en el conducto cístico fue una media de 3.8 minutos y el tiempo de fluorescencia del colédoco fue una media 5.67 minutos. Conclusión: Esta técnica proporciona una visibilidad superior de la anatomía biliar en tiempo real, lo que reduce significativamente el riesgo de lesiones iatrogénicas y mejora la precisión quirúrgica, especialmente en aquellos casos más complejos donde la identificación exacta de las estructuras anatómicas es crucial para evitar complicaciones graves.

This research deals with a series of cases performed prospectively and observationally, to establish the efficacy of the use of intra gallbladder indocyanine green fluorescence in 15 patients undergoing laparoscopic cholecystectomy at the Chair of Clinical and Surgical Therapeutics A, Surgery Service I of the University Hospital of Caracas between June and August 2024. All patients were administered 2.5 mg of indocyanine green, the 25mg ampoule of indocyanine green Aldagor® was diluted in 10 mlof physiological saline solution, intravesicularly with a 25G spinal needle using an OptoMedic® FloNavi 214k laparoscopic tower. The technique was applied in a standardized way in all cases, before dissection of Calot's triangle.Results: Fluorescence was achieved in all cases, in the fundus and body of the gallbladder after intravesicular indocyanine green infiltration, fluorescence occurred at 2.4 minutes, the fluorescence time in the cystic duct was a mean of 3.8 minutes, and the fluorescence time in thecommon bile duct was a mean of 5.67 minutes. Conclusion: This technique provides superior visibility of the biliary anatomy in real time, which significantly reduces the risk of iatrogenic injury and improves surgical precision, especially in more complexcases where accurate identification of anatomical structures is crucial to avoid serious complications.

Colgajo fasciocutáneo perforante de arteria mamaria interna contralateral en reconstrucción de región supraclavicular y evaluación por fluorescencia / Fasciocutaneous perforator flap of the contralateral internal mammary artery in reconstruction of the supraclavicular region, and fluorescence evaluation

Se presenta una reconstrucción posresección tumoral en región supraclavicular en un paciente de 75 años con un colgajo perforante correspondiente a la 2da y 3era perforante intercostal de la mamaria interna contralateral (IMAP, según la nomenclatura de colgajos perforantes). La importancia de ser contralateral a la lesión rompe el dogma respecto a la reconstrucción con colgajos perforantes y su relación con la línea media. Se utilizaron mediciones con verde de indocianina con fluorescencia y dosajes de glucosa para evaluar vitalidad y perfusión del colgajo. Los resultados funcionales y estéticos fueron aceptables.

A reconstruction following a post-tumoral resection in the supraclavicular region is presented in a 75- year- old patient using a perforator fl ap corresponding to the 2nd and 3rd intercostal perforators of the contralateral internal mammary artery (IMAP, according to the perforator fl aps nomenclature). The signifi cance of being contralateral to the lesion breaks the dogma regarding reconstruction with perforator fl aps and their relationship to the midline. Indocyanine green fluorescence measurements and glucosa dosing were used to assess the fl aps vitality and perfusion. The functional and aesthetic results were acceptable.

Fungal luminescence pathways: research and applications / 生物工程学报

The fungal bioluminescence pathway (FBP) is a metabolic pathway responsible for the generation of bioluminescence derived from fungi. This pathway utilizes caffeic acid as the substrate, generating a high-energy intermediate, and the decomposition of which yields green fluorescence with a wavelength of approximately 520 nm. The FBP is evolutionally conserved in luminescent fungal groups. Unlike other bioluminescent systems, the FBP is particularly suitable for engineering applications in eukaryotic organisms, especially in plants. Currently, metabolically engineered luminescent plants are able to emit visible light to illuminate its surroundings, which can be visualized clearly in the dark. The fungal bioluminescent system could be explored in various applications in molecular biology, biosensors and glowing ornamental plants, and even green lighting along city streets.

Evaluación del recolector de orina en la toma de muestra de líquido seminal y tinción diferencial de fluorescencia en espermatozoides / Evaluation of the urine collector in the collection of the seminal fluid sample and differential fluorescent staining method in sperm

Objetivo: Verificar que los recolectores de orina no son tóxicos para los espermatozoides. Métodos: Se evaluó la toxicidad de recipientes de material de vidrio (control) y plástico (recolector de orina) con muestras de semen de buena concentración y movilidad de espermatozoides, mediante el estudio de la movilidad espermática a intervalos de 1 hora, a temperatura ambiente, durante 4 horas, siguiendo lineamientos de la quinta edición del Manual de la Organización Mundial de la Salud para el procesamiento de muestras de semen. Se comparó la movilidad progresiva entre ambos tipos de recipientes (vidrio y plástico) y se evaluaron las características morfotintoriales de los espermatozoides con la tinción diferencial de fluorescencia modificada. Resultados: No se hallaron diferencias estadísticamente significativas (p = 0,334) entre los grupos. Los espermatozoides que emitieron una fluorescencia verde, se categorizaron como normales y presentaron una fuerte correlación (r = 1,000; p = 0,000); mientras que, las tinciones amarilla (r = -0,838, p = 0,009) y anaranjada (r = 0,940, p = 0,000), estuvieron altamente correlacionadas con anormalidades de los espermatozoides, destacándose que la fluorescencia anaranjada, tuvo una alta correlación (r = 0,940, p = 0,000) con anormalidades de la cabeza. Conclusión: El recolector de plástico no tiene efectos tóxicos sobre los espermatozoides, satisface las necesidades de recogida de la muestra de semen. Con la tinción diferencial de fluorescencia modificada, la tinción verde se asocia a espermatozoides morfológicamente normales y las amarilla y anaranjada se asocian a anormalidades de la cabeza, pieza intermedia y pieza principal de los espermatozoides(AU)

Objective: To verify that urine collectors are not toxic to sperm. Methods: The toxicity of glass (control) and plastic (urine collector) containers with semen samples of good concentration and sperm motility was evaluated by studying sperm motility at intervals of 1 hour, at room temperature, for 4 hours, following guidelines of the fifth edition of the World Health Organization Manual for the processing of semen samples. The progressive motility between both types of containers (glass and plastic) was compared and the morphotintorial characteristics of the spermatozoa were evaluated with the modified fluorescence differential stain. Results: No statistically significant differences (p = 0.334) were found between the groups. Spermatozoa that emitted a green fluorescence were categorized as normal and presented a strong correlation (r = 1,000; p = 0.000); Whereas, yellow (r = -0.838, p = 0.009) and orange (r = 0.940, p = 0.000) stains were highly correlated with sperm abnormalities, highlighting that orange fluorescence had a high correlation (r = 0.940, p = 0.000) with head abnormalities. Conclusion: The plastic collector has no toxic effects on sperm, it satisfies the needs of collection of the semen sample. With modified fluorescence differential staining, green staining is associated with morphologically normal spermatozoa and yellow and orange stains are associated with abnormalities of the head, middle piece, and main piece of sperm(AU)

Photosynthetic gas exchange and chlorophyll fluorescence of female hemp plants during sexual reversal treatments / Intercambio gaseoso y fluorescencia de la clorofila en plantas femeninas de Cannabis durante tratamientos de reversión sexual

Sexual reversal methods are commonly used in plant breeding programs, allowing male flowers from female plants or vice versa. This work evaluated sexual reversal methods in female Cannabis plants and their effect on gas exchange activity. Plants treated with 1, methyl-cyclopropene (1-MCP), and aminoetoxyvinylglycine (AVG) showed differences in net photosynthesis (A) and stomatal conductance (gs) between the periods before and after sexual reversal treatments. Quantum yield (Qy), electron transport rate (ETR), and non-photochemical quenching (NPQ) did not show a relationship to the treatments, an increase in Qy and ETR, and a reduction in NPQ were observed after applying treatments. 1-MCP, AVG, and STS (silver thiosulfate) were effective in sexual reversal, while photoperiod changes did not induce the formation of male flowers. Induction of sexual reversion in Cannabis plants did not generate variations in energy dissipation mechanisms through photosystems.

Los métodos de reversión sexual se utilizan comúnmente en los programas de fitomejoramiento, permitiendo la formación de flores masculinas a partir de plantas femeninas y viceversa. Este trabajo tuvo como objetivo evaluar métodos de reversión sexual en plantas femeninas de Cannabis y su efecto sobre el intercambio de gases. Plantas tratadas con 1-metil-ciclopropano (1-MCP) y aminoetoxivinilglicina (AVG) mostraron diferencias en fotosíntesis neta (A) y conductancia estomática (gs) entre los periodos antes y después de los tratamientos de reversión sexual. El rendimiento cuántico (Qy), la tasa de transporte de electrones (ETR) y la disipación no fotoquímica (NPQ) no mostraron relación con los tratamientos, se observó un incremento en Qy y ETR y una reducción en NPQ después de la aplicación de los tratamientos. 1-MCP, AVG y STS (tiosulfato de plata) fueron efectivos en la reversión sexual, mientras que los cambios en el fotoperiodo no indujeron la formación de flores masculinas. La inducción de la reversión sexual en plantas de Cannabis no generó variaciones en los mecanismos que disipan la energía a través de los fotosistemas.

Comparison Between Radiographs, White and Fluorescent Images in the Diagnosis and Treatment Decisions for Occlusal Caries: An Ex Vivo Study

ABSTRACT Objective: To compare the agreement of images in white light (WL), fluorescence (FL), and digital radiographs (DR), on the diagnosis and treatment decisions for occlusal caries lesions against a micro-CT gold standard. Material and Methods: Ten extracted third molars, with enamel and/or dentin caries (ICDAS 2-4), were included. Occlusal surface images were acquired with an intraoral camera (SoproLife®) in WL and FL modes. DR was obtained using an intraoral X-ray and a semi-direct digital system. A total of 780 images were needed, organized in a template, to be later examined by twenty-six dentists invited to compose the study. The Generalized Estimation Equations model was used to compare the proportions of the correct answers between the three methods and the gold standard. When significant, Bonferroni post-hoc test was used to identify differences (α=5%). Results: Most of the examiners were specialists (76.9%) with 14.5 years of experience. All diagnostic methods were similar and showed low agreement (DR 12.7%, WL 16.5%, and FL 16.5%) compared with gold standard caries diagnostic scores. Regarding treatment decisions, mean agreement for all diagnostic methods was higher (43.2%; p<0.001), and among all methods, WL (48.1%) and FL (51.2%) modes performed better than DR (30.4%, p<0.001). Conclusion: SoproLife® images could help clinicians to propose rational, minimally invasive treatments for occlusal caries lesions.

Ação da luz visível na oxidação da melanina e no descolorimento do cabelo / Action of the visible on the melanin oxidation and hair discoloration

O dano capilar causado pelo descolorimento oxidativo é muito intenso, sendo que dois fatores são responsáveis por essa ação: primeiro, a ação direta e danosa do oxidante em diversas estruturas capilares e segundo, o dano oxidativo primário facilita o dano causado por outros agentes físicos (luz, temperatura) e químicos (tensoativos), que comumente tem ação nos cabelos. Desenvolver conceitos e tecnologias que possam tornar o oxidante específico para a melanina e por conseguinte efetuando o descolorimento sem causar danos ao fio é extremamente desejável. Neste trabalho buscaremos entender de que forma a luz visível pode aumentar a ação do oxidante sem danificar o fio colateralmente. O objetivo principal deste trabalho é demonstrar que é possível utilizar a luz visível, que é absorvida pela melanina, para tornar esse pigmento mais suscetível ao agente oxidante e desta forma, permitir que o descolorimento seja realizado com concentrações pequenas de oxidante. Também almejamos desenvolver métodos de análises por microscopia ótica de fluorescência e de reflexão para mensurar o dano nas estruturas dos fios processados com oxidante e na presença ou ausência da luz

The capillary damage caused by oxidative discoloration is very intense, and two factors are responsible for this action: first, the direct and harmful action of the oxidant on several capillary structures and second, the primary oxidative damage facilitates the damage caused by other physical agents (light, temperature) and chemicals (surfactants), which commonly have action on the hair. Developing concepts and technologies that can make the oxidant specific to melanin and therefore discoloring without causing damage to the hair is extremely desirable. In this work we will try to understand how visible light can increase the oxidant's action without damaging the wire collaterally. The main objective of this work is to demonstrate that it is possible to use visible light, which is absorbed by melanin, to make this pigment more susceptible to the oxidizing agent and, thus, to allow the discoloration to be carried out with small concentrations of oxidizer. We also aim to develop methods of analysis by optical fluorescence and reflection microscopy to measure the damage to the structures of the threads processed with oxidizer and in the presence or absence of light

Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope / 中国中药杂志

A fluorescence endoscopic laser confocal microscope(FELCM) was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN) into the joint, and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA) was evaluated. Sin-SLN was prepared with the emulsion evaporation-low temperature curing method. The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%, the drug loading of 3.84%±0.28%, the average particle size of(215.27±4.21) nm, and the Zeta potential of(-32.67±0.84) mV. Moreover, the Sin-SLN demonstrated good stability after sto-rage for 30 days. The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant. Five groups were designed, including a control group, a model group, a Sin(1.5 mg·kg~(-1)) group, a Sin-SLN(1.5 mg·kg~(-1)) group, and a dexamethasone(positive drug, 1.0 mg·kg~(-1), ig) group. The control group and the model group only received puncture treatment without drug injection. After drug administration, the local skin temperature and knee joint diameter were monitored every day. The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05, P<0.01). FELCM recorded the morphological alterations of the cartilage of knee joint. The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the serum le-vels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α). The findings revealed that the Sin-SLN group had lower IL-1 and TNF-α levels than the model group(P<0.05, P<0.01). Hematoxylin-eosin(HE) staining was employed to reveal the pathological changes of the synovial tissue, which were significantly mitigated in the Sin-SLN group. The prepared Sin-SLN had uniform particle size and high stability. Through joint injection administration, a drug reservoir was formed. Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbit, down-regulated the expression of inflammatory cytokines, and inhibited the epithelial proliferation and inflammatory cell infiltration of the synovial tissue, demonstrating the efficacy in treating RA.

Research Progress of Indocyanine Green Fluorescence Laparoscopic Technique in Clinical Application / 中国医疗器械杂志

Indocyanine green (ICG) is the most commonly used near-infrared fluorescent (NIRF) dye in clinical practice, and its mediated near-infrared fluorescence imaging technology is gradually applied in clinical practice. It has shown great potential in invasive surgery (MIS) and is expected to become the standard technology for surgical diagnosis and treatment of diseases. The clinical application of ICG fluorescence laparoscopy is reviewed here.

Uso de verde de indocianina en cirugía reconstructiva y quemaduras / The use of indocyanine green in reconstructive surgery and burns: the literature review

Resumen El verde de indocianina es un tinte que se ha utilizado en medicina durante varias décadas. Tiene una serie de aplicaciones, incluida la cirugía reconstructiva y las quemaduras. Permite detectar áreas de tejido con perfusión reducida, lo que reduce el riesgo de complicaciones posoperatorias en forma de procesos de cicatrización alterados y necrosis. La técnica de imágenes que utiliza este tinte, permite observar los cambios en la fluorescencia en tiempo real y que, se ha demostrado, ocurren entre las capas superficiales y profundas en las quemaduras. Esto permite un diagnóstico cualitativo y cuantitativo de la profundidad de la quemadura, lo que se traduce en la elección de un tratamiento adicional. Se aprecia la importancia particular de este método en la prevención de la necrosis cutánea con el complejo areola-pezón durante la reconstrucción mamaria simultánea. Se necesitan más ensayos controlados aleatorios prospectivos para considerarlo el "método de elección" en la práctica clínica.

Indocyanine green is a dye that has been used in medicine for several decades. It has a number of applications, including reconstructive surgery and burns. It allows the detection of areas of tissue with reduced perfusion, which reduces the risk of postoperative complications in the form of altered healing processes and necrosis. The imaging technique that uses this dye allows us to observe the changes in fluorescence in real time that have been shown to occur between the superficial and deep layers in burns. This allows a qualitative and quantitative diagnosis of the depth of the burn, which results in the choice of additional treatment. The particular importance of this method in the prevention of skin necrosis with the areolanipple complex is appreciate during simultaneous breast reconstruction. More prospective randomized controlled trials are needed to consider it the 'method of choice' in clinical practice.

Resección Ósea Guiada por Fluorescencia en Pacientes con Osteonecrosis Mandibular Asociada a Medicamentos: scoping Review / Fluorescence-Guided Bone Resection in Patients With Drug-Associated Osteonecrosis of the Mandible: scoping Review

RESUMEN: La osteonecrosis mandibular asociada a medicamentos (MRONJ) es una enfermedad grave, pero de baja prevalencia en la cual los márgenes quirúrgicos de la resección ósea son determinados clínicamente por el cirujano basado en parámetros subjetivos. Debido a esto, se ha propuesto la resección ósea guiada por fluorescencia. Describir el conocimiento act ual sobre la resección ósea guiada por fluorescencia en pacientes con MRONJ. Se realizó Scoping Review, buscando en las bases de datos de Pubmed, ScienceDirect y Web of Science, utilizando los operadores booleanos AND y OR. Se aplicaron criterios de inclusión y exclusión. De 66 artículos iniciales, se seleccionaron 13 que cumplieron con los criterios de elegibilidad. Actualm ente se utiliza la técnica de fluorescencia con tetraciclinas (FT) y autofluorescencia (AF), describiéndose un éxito del 85 a 100 % y 7 7,3 a 100 % en un tiempo de seguimiento 1 a 12,9 y 4 a 13,1 meses respectivamente. Hubo 100 % de correlación Clínico-Histopatológica para tejido óseo vital y necrótico. La resección ósea guiada por fluorescencia logra discriminar clínicamente entre tejido óseo vital y necrótico, delimitando los márgenes quirúrgicos de la resección y/o desbridamiento basado en parámetros objetivos y reproducibl es que permiten estandarizar el procedimiento quirúrgico.

ABSTRACT: Medication-related Osteonecrosis of the jaw (MRONJ) is a serious but low prevalence disease in which the surgical margins of bone resection are clinically determinated by the surgeon based on subjetive parameters. Because of this, fluorescence-guided bone resection has been proposed. The objetive of this study is to describe the current knowledge of fluorescence-guided bone resection in patients with MRONJ. Scoping Review was carried out, searching the databases of PubMed, ScienceDirect and Web of Science, using the boolean operator AND and OR. Inclusion and exclusion criteria were applied. Of 66 articles, 13 were selected that met the eligibility criteriaThe technique of fluorescence with tetracyclines (FT) and autofluorescence (AF) is used, describing a success of 85 to 100 % and 77.3 to 100 % in 1 to 12.9 and 4 to 13.1 months respectively. There was 100 % Clinical-Histopathological correlation for vital and necrotic bone tissue. Fluorescence-guided bone resection is able to clinically discriminate between vital and necrotic bone tissue, allowing the delimitation of surgical margins based on objective and reproducible parameters. In addition, it allows to standardize the surgical procedure.

Esofagectomía con linfadenectomía guiada por fluorescencia con verde de indocianina en cáncer de esófago. Experiencia inicial en la Argentina / Esophagectomy with lymphadenectomy guided by fluorescence imaging with indocyanine green in esophageal cancer. Initial experience in Argentina

RESUMEN Antecedentes: en la última década ha comenzado a investigarse el uso de la captación de fluorescencia mediante luz infrarroja para la visualización de ganglios linfáticos en tumores de estómago y esófago. Objetivo: evaluar la factibilidad de la evaluación del drenaje linfático de cáncer de esófago mediante el uso de fluorescencia y verde de indocianina (ICG). Material y métodos: se realizó un estudio prospectivo que incluyó pacientes con tumores de la unión gastroesofágica resecables (estadios I, II y III). Antes de comenzar la cirugía se inyectaron por endoscopia 4 mL de ICG doblemente diluida en agua estéril (1,25 mg/mL = 5 mg) en la submucosa del esófago en los cuatro cuadrantes (1 mL por cuadrante) alrededor del tumor. Resultados: se incluyeron en total 6 pacientes. En todos ellos se logró identificar el drenaje linfático del tumor hacia la primera estación ganglionar: en 6/6 (100%), el drenaje linfático con fluorescencia se detectó en las estaciones ganglionares N°s 3 y 7 (curvatura menor y arteria gástrica izquierda). En ningún paciente se identificó fluorescencia en ganglios mediastinales. Conclusión: la visualización del drenaje linfático de tumores de la unión gastroesofágica mediante el uso de fluorescencia con ICG es factible.

ABSTRACT Background: Over the past decade, fluorescence imaging with infrared light has been used to visualize lymph nodes in tumors of the stomach and esophagus. Objective: The aim of our study was to evaluate the feasibility of evaluating lymphatic drainage in esophageal cancer using fluorescence and indocyanine green (ICG). Material and methods: We conducted a prospective study of patients with resectable tumors of the gastroesophageal junction (stage I, II and III). Before surgery, 4 mL of ICG double diluted in sterile water (1.25 mg/mL = 5 mg) were injected via endoscopy into the esophageal submucosa in the four quadrants (1 mL per quadrant) around the tumor. Results: A total of 6 patients were included. Lymphatic drainage from the tumor to the first lymph node station was identified in all patients: in 6/6 (100%), fluorescent lymphatic drainage was detected in nodal stations number 3 and 7 (lesser curvature and left gastric artery) Fluorescence was not identified in the mediastinal lymph nodes in any patient. Conclusion: Visualization of lymphatic drainage of gastroesophageal junction tumors to the lesser curvature nodes using fluorescence imaging is feasible.

Ecophysiology and water use efficiency of soybean cultivars under field conditions

A wide range of soybean cultivars is available on the market and understanding the physiological response and yield of these materials is fundamental to develop new management systems. Thus, the objective of the present study was to assess ecophysiological parameters and yield of soybean cultivars under field conditions. The experiment was carried out on a farm located in the municipality of Açailândia, Maranhão, Brazil. Three commercial cultivars were used (SC1, SC2 and SC3), and gas exchanges, SPAD index, Fv/Fm, photosynthesis index (PI), instantaneous water use efficiency (WUE) and intrinsic instantaneous of the use of water (iWUE) were assessed during the vegetative (V5) and reproductive (R5) stages. In addition, the biomass and production components were obtained. A randomized complete block design was used, with three cultivars and six replications. SC2 obtained the best mean for the photochemical variables. SC2 was more efficient at both development stages in WUE, but the maximum iWUE values were obtained in SC3. The SC2 cultivar obtained the best responses in the main variables analyzed, resulting in a higher yield.

Estudo da acurácia de dispositivos de indução da fluorescência e do método radiográfico para o diagnóstico de lesões de cárie em dentes decíduos: uma revisão sistemática e meta-análise / Assessment of the accuracy of the fluorescence devices and radiography for the diagnosis of caries in primary teeth: a systematic review and meta-analysis

Objetivo: O objetivo dessa revisão sistemática e meta-análise foi avaliar a acurácia de dispositivos de indução de fluorescência e do método radiográfico para diagnóstico da cárie dentária em dentes decíduos. Métodos: Buscas no PubMed, Web of Science, Scopus, Embase, Open Grey e Google Scholar foram realizadas em agosto de 2021. Referências que preenchiam os critérios de elegibilidade foram incluídas. O risco de viés dos artigos incluídos foi avaliado com a ferramenta QUADAS-2. Foram realizadas meta-analises com o MedCalc. Resultados fornecidos em área sob a curva (ASC), intervalo de confiança (IC) e erro padrão (EP). Resultados: Títulos/resumos de 619 referências foram avaliados por dois revisores calibrados de forma independente. Cinquenta e seis artigos foram selecionados para leitura do texto completo. Onze artigos foram incluídos na revisão-sistemática e meta-análise. Os artigos incluídos apresentaram baixo risco de viés. A ASC para diagnóstico de cárie em esmalte com o laser DIAGNOdent foi de 75,8% (IC = 68,1% - 83,6% / EP = 3,9% / I2 = 67,95%) e para o diagnóstico de cárie em dentina foi de 80,6% (ASC = 80,6% / IC = 65,7% - 95,6% / EP = 7,6% / I2 = 89,24%). A ASC para o diagnóstico de cárie em esmalte com a radiografia interproximal foi de 61,7% (IC = 51,4% - 72,1% / EP = 5,3% / I2 = 44,51%) e a ASC para diagnóstico de cárie em dentina com a radiografia interproximal foi de 73,7% (IC = 45,1% - 100% / EP = 14,6% / I2 = 96,97%). Conclusão: O método radiográfico e os dispositivos de indução de fluorescência são boas opções para diagnóstico de cárie em dentes decíduos. Ambos apresentam melhores resultados em lesões em dentina. Os métodos que utilizam fluorescência tiveram melhores resultados em esmalte quando comparados à radiografia.

Aim: This systematic review and meta-analysis aimed to evaluate the accuracy of the fluorescence induction devices and radiographic method for diagnosing dental caries in primary teeth. Methods: Searches in PubMed, Web of Science, Scopus, Embase, Open Grey, and Google Scholar were conducted in August 2021. References that met the eligibility criteria were included. The risk of bias of the included articles was assessed with QUADAS-2. Meta-analyses were performed with MedCalc. Results were provided as area under the curve (AUC), confidence interval (CI), and standard error (SE). Results: Titles/abstracts of 619 references were evaluated by two trained reviewers independently. Fifty-six articles were selected for full-text reading. Eleven articles were included in the systematic review and meta-analysis. The included articles showed a low risk of bias. The AUC for the diagnosis of caries on enamel with the DIAGNOdent laser was 75.8% (CI = 68.1% - 83.6% / SE = 3.9% / I2 = 67.95%), whereas for the diagnosis of caries on dentin, it was 80.6% (AUC = 80.6% / CI = 65.7% - 95.6% / SE = 7.6% / I2 = 89.24%). The AUC for the diagnosis of enamel caries with the interproximal radiograph was 61.7% (CI = 51.4% - 72.1% / SE = 5.3% / I2 = 44.51%) and the AUC for the diagnosis of dentin caries with the interproximal radiograph was 73.7% (CI = 45.1% - 100% / SE = 14.6% / I2 = 96.97%). Conclusion: Radiography and fluorescence devices are useful tools for the diagnosis of caries in deciduous teeth. Both showed better results in dentin lesions. The methods using fluorescence had better results in enamel when compared to radiography.

Optimization of expression yield in a stable cell line expressing a novel mutated chimeric tissue plasminogen activator (mt-PA)

Abstract The development of stable cell lines producing recombinant proteins is very time-consuming and laborious. One of the practical approaches successfully performed is Fluorescence-Activated Cell Sorting (FACS). A mutated chimeric tissue plasminogen activator (mt-PA) was developed by removing the first three domains of t-PA, insertion of GHRP sequence and mutation toward resistance to plasminogen activator inhibitor-1 (PAI-1). In the current study, a new stable CHO-DG44 cell line producing mt-PA was developed by two sequential clonal selections: FACS and clonal-selection by limiting dilution. Furthermore, the expression was more evaluated using two different expression media. Finally, the high-producing clones were selected based on the dot blot and amidolytic activity test. The transfection efficiency of CHO-DG44 cells was 38% as measured by flow cytometry on green fluorescent protein (GFP). After performing FACS on stable cell pools, the expression yield was increased to fifty-fold. In terms of growth profile, CD-DG44 showed higher viability and cell density results than ProCHO5 medium. The expression of mt-PA was significantly higher in CD-DG44 than in ProCHO5, 765 and 280 IU/mL, respectively. Our data indicated that selection of an appropriate expression medium played a critical role in the development of potent producing stable cells by FACS.

A Selective Fluorescent Probe for the Determination and Imaging of Fluoride in Living Cells

Abstract Fluoride anions are indispensable trace elements required for sustaining life. To investigate the homeostasis and action of fluoride in the body, a new highly sensitive and selective fluorescence detection method was designed for fluoride in aqueous solutions. A fluorescent probe for fluoride (FP-F) was synthesized for imaging F- in living cells. The design strategy for the probe was based on the specific reaction between fluoride and silica to mediate deprotection of this probe to fluorescein. Upon treatment with F-, FP-F, a closed and weakly fluorescent lactone, was transformed into an open and strongly fluorescent product. Under the optimum conditions, the detection limit for fluoride was 0.526 nM. FP-F could detect micromolar changes in F- concentrations in living cells by confocal microscopy.

Quantitative evaluation of the regenerated dura mater in a decompressive skull trepanation model in rats / Evaluación cuantitativa de la duramadre regenerada en un modelo de perforación cranial descompresiva en ratas

SUMMARY: Regeneration of the dura mater following duraplasty using a collagen film, a chitosan film, or a combination of both with gelatin, was studied in a craniotomy and penetrating brain injury model in rats. Collagen autofluorescence in the regenerated dura mater was evaluated using confocal microscopy with excitation at λem = 488 nm and λem = 543 nm. An increase in regeneration of the extracellular matrix of connective tissue and an increase in matrix fluorescence were detected at 6 weeks after duraplasty. The major contributors to dura mater regeneration were collagen films, chitosan plus gelatin-based films, and, to a much lesser extent, chitosan-based films. By using autofluorescence densitometry of extracellular matrix, the authors were able to quantify the degree of connective tissue regeneration in the dura mater following duraplasty.

RESUMEN: Se estudió la regeneración de la duramadre después de una duraplastía utilizando una lámina de colágeno, una lamina de quitosano o una combinación de ambas con gelatina en un modelo de craneotomía y lesión cerebral en ratas. La autofluorescencia del colágeno en la duramadre regenerada se evaluó mediante microscopía confocal con excitación a λem = 488 nm y λem = 543 nm. Se observó un aumento en la regeneración de la matriz extracelular del tejido conectivo y un aumento en la fluorescencia de la matriz a las 6 semanas después de la duraplastía. Se observe un efecto significativo en la regeneración de la duramadre con las láminas de colágeno, las láminas en base de quitosano más gelatina y, en un menor grado, las láminas a base de quitosano. Mediante el uso de densitometría de autofluorescencia de la matriz extracelular, los autores lograron cuantificar el grado de regenera- ción del tejido conectivo en la duramadre después de la duraplastía.

Uso excesivo de la prueba de ANCA / Overuse of the neutrophil cytoplasmic antibody test

La desproporcional y alta frecuencia de órdenes médicas de anticuerpos frente al citoplasma del neutrófilo (ANCA, por sus siglas en inglés) dirigidas a nuestros laboratorios clínicos evidencia el sobreuso de la prueba de ANCA. El uso indiscriminado de esta aumenta los gastos sin beneficio de salud. El laboratorio clínico es el eslabón de la cadena diagnóstica que más siente el uso excesivo de las solicitudes de ANCA, básicamente porque genera resultados falsos positivos que comprometen la utilidad clínica de la prueba, además de recargar innecesariamente el trabajo diario del laboratorio. La prueba de ANCA es una herramienta diagnóstica muy útil para las vasculitis sistémicas primarias, pero su valor en situaciones no vasculíticas así como en otras condiciones inflamatorias y en enfermedades infecciosas o tumorales, no ha sido demostrado.1,2 El descubrimiento de los ANCA cambió...(AU)

Uso de un nuevo protocolo clínico fotográfico-computacional en el registro de la fluorescencia visible inducida por radiación ultravioleta en dientes humanos / Use of a new clinical photographic-computational protocol to register the ultraviolet-induced visible fluorescence in human teeth

RESUMEN: Los dientes humanos experimentan fluorescencia visible al ser sometidos a radiación ultravioleta, la cual varía de color a medida que el diente envejece. Objetivo: el presente estudio buscó desarrollar un protocolo clínico sencillo usando fotografía digital y análisis computacional para determinar la fluorescencia visible inducida por radiación ultravioleta (FVIUV) en dientes anteriores de pacientes de distintos grupos etarios. Materiales y Métodos: 35 participantes de 5 grupos etarios fueron reclutados. 70 incisivos centrales superiores fueron fotografiados utilizando como fuente de iluminación luz UV. Cada imagen obtenida fue analizada con el software computacional ImageJ para obtener las coordenadas de color en el espacio CIELab para la corona completa. Los datos obtenidos fueron analizados mediante las pruebas estadísticas de ANOVA y post-hoc de Tukey HSD. Resultados: Se obtuvieron las magnitudes de FVIUV en las tres dimensiones del espacio de color CIELab en la corona completa. Esta FVIUV se manifestó en el espectro del color verdeazulado para todos los grupos etarios, los cuales presentaron diferencias de saturación y luminosidad según tramo etario. Conclusiones: El protocolo aplicado en presente estudio permitió la cuantificación de la FVIUV dentaria en distintos grupos etarios.

ABSTRACT: Human teeth experience visible fluorescence when subjected to ultraviolet radiation, which varies in color as the tooth ages. Objective: The present study sought to develop a simple clinical protocol using digital photography and computational analysis to determine the difference in visible fluorescence induced by ultraviolet radiation (FVIUV) in anterior teeth of patients of different age groups. Materials and Methods: 35 participants from five age groups were recruited. 70 upper central incisors were photographed using UV light as the illumination source. Each image obtained was analyzed with ImageJ computer software to obtain the color coordinates in CIELab space for the entire crown. The data obtained was analyzed using the ANOVA and Tukey HSD post-hoc statistical tests. Results: Visible fluorescence magnitudes were obtained in the three dimensions of the CIELab color space in the complete crown. This FVIUV was registered in the spectrum of the blue-green color for all age groups, which presented differences in saturation and luminosity according to age group. Conclusions: The protocol applied in this study allowed the quantification of dental FVIUV in different age groups.

Comparative performances of seven quantitative Reverse - Transcription Polymerase Chain Reaction assays (RT-qPCR) for detecting SARS-CoV-2 infection in samples from individuals suspected of COVID-19 in São Paulo, Brazil

Introduction Brazil is the second largest country with COVID-19 positive cases worldwide. Due to the potent spread of the virus and the scarcity of kits and supplies, the Brazilian Ministry of Health has granted authorization for the use of kits available during this emergency, without an accurate evaluation of their performance. This study compared the performance and cost-effectiveness of seven molecular assays/kits available in São Paulo, Brazil, for SARS-CoV-2 diagnosis Materials and methods A total of 205 nasopharyngeal/oropharyngeal samples from suspected cases of COVID-19, were tested using the following assays: (i) GeneFinder COVID-19 plus RealAmp kit; (ii) 2019-nCoV RNA PCR-Fluorescence Probing, Da An Gene Co.; (iii) in-house RT-qPCR SARS-CoV-2 IAL; (iv) 2019-nCoV kit, IDT; (v) molecular SARS-CoV-2 (E) kit, Bio-Manguinhos; (vi) Allplex 2019-nCoV modified Assay, Seegene Inc, and (vii) Biomol one-step COVID-19 kit, IBMP. The criteria for determining a SARS-CoV-2 true positive result included the cycle threshold cut-off values, the characteristics of exponential/linear curves, the gene target diversity, and a positive result in at least two assays Results The overall sensitivity of the assays listed were GeneFinder 83.6%, Da An Gene 100.0%, IAL 90.4%, IDT 94.6%, Bio-Manguinhos 87.7%, Allplex 97.3%, and IBMP 87.7%. The minor sensitive gene target was RdRP. Although all assays had a Cohen's Kappa index ≥0.893, the best tests used multiplex assays identifying N-gene and/or E-gene targets Conclusion All assays tested accurate for diagnosis, but considering cost-effectiveness (cost, time consumption, number of samples tested, and performance), the in-house IAL assay was ideal for COVID-19 diagnosis in São Paulo, Brazil.