Cellulose acetate microwell plates for high-throughput colorimetric assays.

Single use plasticware (SUP) in scientific, diagnostic, and academic laboratories makes a significant contribution to plastic waste generation worldwide. Polystyrene (PS) microwell plates form a part of this waste. These plates are the backbone of high throughput colorimetric measurements in academic, research, and healthcare settings for detection/quantification of wide-ranging analytes including proteins, carbohydrates, nucleic acids, and enzyme activity. Polystyrene (PS) microwell plates serve as a platform for holding samples and reagents, where mixing initiates chemical reaction(s), and the ensuing color changes are quantified using a microplate reader. However, these plates are rarely reused or recycled, contributing to the staggering amounts of plastic waste generated in scientific laboratories. Here, we are reporting the fabrication of cellulose acetate (CA) microwell plates as a greener alternative to non-biodegradable PS plates and we demonstrate their application in colorimetric assays. These easy to fabricate, lighter weight, customizable, and environmentally friendly plates were fabricated in 96- and 384-well formats and made water impermeable through chemical treatment. The plates were tested in three different colorimetric analyses: (i) bicinchoninic acid assay (BCA) for protein quantification; (ii) chymotrypsin (CT) activity assay; and (iii) alkaline phosphatase (AP) activity assay. Color intensities were quantified using a freely available smartphone application, Spotxel® Reader (Sicasys Software GmbH). To benchmark the performance of this platform, the same assays were performed in commercial PS plates too and quantified using a UV/Vis microplate reader. The two systems yielded comparable linear correlation coefficients, LOD and LOQ values, thereby validating the CA plate-cell phone based analytical method. The CA microwell plates, coupled with smart phone optical data capture, provide greener, accessible, and scalable tools for all laboratory settings and are particularly well-suited for resource- and infrastructure-limited environments.

Safety and tolerability of sedation-free flexible cystoscopy for intradetrusor botulinum toxin-A injection.

PURPOSE: Intradetrusor botulinum toxin-A injection is a promising emerging therapy for neurogenic and idiopathic overactive bladder that is refractory to current antimuscarinic agents. We evaluated a sedation-free procedure using flexible endoscopy to perform intradetrusor botulinum toxin-A injection using a validated pain score to assess the tolerability of this technique. MATERIALS AND METHODS: Five men and 22 women with idiopathic overactive bladder refractory to antimuscarinics were prospectively evaluated and randomized to 100 U (14) or 150 U botulinum toxin-A (13). Subjects underwent BTA injection with intravesical lidocaine using a 14Fr Olympus flexible cystoscope, which accommodates a 27 gauge flexible Olympus injection needle. Patients were evaluated with the visual analog scale to evaluate discomfort during and 15 minutes after the procedure. Procedure time was recorded. RESULTS: In 22 female patients the mean pain score was 3.1 (range 0 to 10) during the procedure and 0.7 (range 0 to 7) 15 minutes after the procedure. In 5 male patients the mean pain score was 1.6 (range 0 to 3.5) during the procedure and 0.0 with all reporting no pain 15 minutes after the procedure. Mean procedure time was 4.5 minutes (range 4 to 6). Only 1 of the 27 patients requested sedation before any subsequent injections. CONCLUSIONS: Sedation-free intradetrusor botulinum toxin-A injection using intravesical lidocaine and flexible endoscopy is a well tolerated and safe procedure to perform in an office setting.

Staging rectourinary fistulas to guide surgical treatment.

PURPOSE: Surgical management for rectourinary fistulas remains a reconstructive challenge. There are few guidelines to direct the surgeon to the most successful and least morbid technique. We developed a rectourinary fistula staging system that allows selection of the most appropriate technique for the patient. We present the details of the staging system and surgical outcomes. MATERIALS AND METHODS: From July 1999 to July 2005 we treated 14 male patients with rectourinary fistula. Mean patient age was 68 years (range 62 to 73). Etiology was rectal injury during open radical prostatectomy in 5 patients, laparoscopic prostatectomy in 1, radiation induced fistula for prostate cancer treatment (brachytherapy and external beam radiation therapy) in 2, neoadjuvant external beam radiation therapy in 2, ischial decubitus ulcer in 3 with spinal cord injury, and cryotherapy and external beam radiation therapy in 1. Cases were staged as stage I--low (less than 4 cm from anal verge and nonirradiated), stage II--high (more than 4 cm from anal verge and nonirradiated), stage III--small (less than 2 cm irradiated fistula), stage IV--large (more than 2 cm irradiated fistula) and stage V--large (ischial decubitus fistula). Diverting colostomy was performed for stages III to V 6 weeks before definitive therapy. RESULTS: Patients were discharged home after 48 hours. A 22Fr urethral catheter maintained bladder drainage for 3 weeks until cystogram confirmed rectourinary fistula closure. Complications were superficial wound infection and postoperative reexploration of the gracilis flap due to bleeding in 1 case each. All patients were cured after a single operation. CONCLUSIONS: The surgical challenges of a variety of rectourinary fistula repairs can be managed with minimal morbidity and a high success rate using proper staging to guide urinary tract reconstruction.

Las secuelas neurológicas de origen perinatal en su relación con los factores de riesgo. Modelo de estudio / Neurological sequels of perinatal origin and factor risk. A model to study

Antecedentes. Se postula la importancia de las secuelas neurológicas de origen perinatal. Se revisa la definición de secuela, insistiendo en considerarla como un proceso que se desencadena a partir de ciertos factores de riesgo que la condicionan, para poder abordar el problema a partir de los factores de riesgo y establecer pogramas que prevengan los síndromes de parálisis cerebral, retardo mental y alteraciones de conducta. Objetivo. Señalar la insuficiencia de los procedimientos estadísticos descriptivos y univariados, y proponer el empleo de modelos complejos de análisis. Material y métodos. Se presentan algunos datos publicados en la literatura relativos a la frecuencia de presentación de secuelas y finalmente se expone el programa de la Clínica de Neurodesarrollo del Instituto Nacional de Pediatría-Universidad Autónoma Metropolitana, Unidad Xochimilco. Resultados. Mediante los procedimientos estadísticos descriptivos y univariados no se comprueba la asociación entre factores de riesgo como peso, condición al nacimiento, edad gestacional y datos del diagnóstico de la encefalopatía por US cerebral, EEG, potenciales evocados, exploración clínica neurológica y los datos de los casos y desarrollo posterior de secuelas. Con análisis complejos sí se demuestra la asociación y se insiste en que no es directa ni lineal