A competitive assay for the detection of a 16-mer peptide from α1 chain of human collagen XI.

Characterization of extracellular matrix (ECM) is becoming more and more important to decipher cancer progression. Constant remodeling results in ECM components degradation or unusual ECM accumulation that releases short fragments to the body fluids. These fragments might be potential cancer biomarkers but to detect them specific receptors are needed. In response to this demand, we present the first electrochemical aptamer-based competitive assay for the minor collagen XI, dysregulated in several carcinomas. It was performed on magnetic beads using enzymatic labeling. First, we selected the most appropriate tag for the aptamer (biotin or 6-carboxyfluorescein). The former yielded higher currents by chronoamperometry and it was used for the competitive assay. The collagen fragment, a 16mer peptide used as the target, was detected from 52 to 1000 nM with an RSD of about 5%. The LOD of the assay was estimated as 24 nM (44 ng/mL). The performance of the assay in serum diluted 1:2 was equivalent to the assay in PBS. The detection of α1 chain of human collagen XI was also possible in cell lysates and confirmed by aptacytofluorescence, which is promising as a new tool to validate this fragment as a cancer biomarker.

The Barcelona-Asymptomatic Intracranial Atherosclerosis (AsIA) study: Subclinical cervico-cerebral stenosis and middle cerebral artery pulsatility index as predictors of long-term incident cognitive impairment.

BACKGROUND AND AIMS: We aimed to study subclinical non-invasive vascular markers as predictors of incident long-term cognitive impairment in a longitudinal population-based study. METHODS: The Barcelona-Asymptomatic Intracranial Atherosclerosis (AsIA) study is a population-based study that included a random sample of 933 Caucasian subjects (mean age 66 years, 64% male) with a moderate-high vascular risk and without history of stroke or dementia. Subclinical carotid and intracranial stenosis was assessed at baseline visit by cervical and transcranial color-coded duplex (TCCD) and confirmed by magnetic resonance angiography. Cervico-cerebral stenosis (CCS) was defined as the presence of extra and/or intracranial stenosis >50%. Baseline middle cerebral artery pulsatility index (MCA-PI) was measured bilaterally by TCCD, and mean PI of both sides was considered for analyses. Subjects were followed-up to determine incident long-term cognitive impairment (mild cognitive impairment or dementia). RESULTS: After a median of 7.16 [6.91-7.75] years of follow-up, 91 subjects (9.7%) developed cognitive impairment, 27 of them mild cognitive impairment, and 64 dementia. Incidence of cognitive impairment was significantly higher among subjects with subclinical CCS (21.4% versus 9% in those without CCS) and among those with mean MCA-PI>1 (13.5% versus 7.4% in those with MCA-PI<1). In multivariate Cox regression analyses, both CCS and MCA-PI>1 were independently associated with incident cognitive impairment with HR of 2.07 [1.11-3.88] and 1.58 [1.02-2.46], respectively. CONCLUSIONS: Subclinical cervico-cerebral stenosis and higher MCA-PI are non-invasive neurosonological markers of incident long-term cognitive impairment in our population.

Aptamers targeting protein-specific glycosylation in tumor biomarkers: general selection, characterization and structural modeling.

Detecting specific protein glycoforms is attracting particular attention due to its potential to improve the performance of current cancer biomarkers. Although natural receptors such as lectins and antibodies have served as powerful tools for the detection of protein-bound glycans, the development of effective receptors able to integrate in the recognition both the glycan and peptide moieties is still challenging. Here we report a method for selecting aptamers toward the glycosylation site of a protein. It allows identification of an aptamer that binds with nM affinity to prostate-specific antigen, discriminating it from proteins with a similar glycosylation pattern. We also computationally predict the structure of the selected aptamer and characterize its complex with the glycoprotein by docking and molecular dynamics calculations, further supporting the binary recognition event. This study opens a new route for the identification of aptamers for the binary recognition of glycoproteins, useful for diagnostic and therapeutic applications.

[Single-repair for primary proximal hypospadias. Lessons learned after 196 primary repairs]. / Hipospadias proximales reparados en un único tiempo. Lecciones aprendidas tras 196 intervenciones primarias.

PURPOSE: Proximal, scrotal and perineal hypospadias challenge the surgeon. After 40 years devoted to hypospadias surgery, the senior author summarizes her experience, share her tricks and shows that almost any hypospadias can be fully repaired in one surgery. METHODS: Retrospective review and discussions of a large series of consecutive proximal hypospadias treated in one surgery by the same team from 1999 to 2016. RESULTS: 196 patients were operated. 68% of all patients were successfully repaired after the surgery. All hypospadias were treated using one of the following techniques: onlay double flap, tubularized preputial flap or mucosal grafts (only until 2005). Urethrocutaneous fistula was present in 25% of cases, but only 13% of patients needed other further surgical procedures. CONCLUSION: Cutaneous incisions design is fundamental in hypospadias repair. When the incisions follow the natural folds that appear in penile and scrotal skin, it's possible to obtain optimal flaps for both urethroplasty and skin coverage (even avoiding the need for mucosal grafts). Consequently, almost all kinds of hypospadias can be repaired in one surgery with very good long-term results.

INTRODUCCION Y OBJETIVO: Los hipospadias peneanos proximales, escrotales y perineales son un reto para el cirujano. Tras toda una carrera dedicada a los hipospadias, la autora principal del trabajo resume su experiencia y demuestra que prácticamente cualquier tipo de hipospadias puede ser resuelto completamente en un único tiempo. METODO: Se trata de una revisión retrospectiva y la discusión de una serie de hipospadias proximales consecutivos tratados en una única intervención por el mismo equipo quirúrgico entre 1999 y 2016. RESULTADOS: 196 pacientes fueron intervenidos. El 68% de los casos quedaron satisfactoriamente resueltos tras la operación. Todos los hipospadias fueron reparados con una de las siguientes técnicas: onlay, colgajo prepucial tubularizado (Duckett) o injertos de mucosa. Los injertos se utilizaron solo hasta 2005; posteriormente, el refinamiento de las técnicas quirúrgicas expuestas permitió que cualquier hipospadias fuera reparado con las técnicas de onlay o Duckett. Solo un 13% de los casos necesitó algún tipo de procedimiento quirúrgico adicional, más allá del cierre de una fístula uretral puntiforme (presente en el 25% de los pacientes). CONCLUSION: El diseño de las incisiones cutáneas es fundamental en la reparación de los hipospadias. Cuando las incisiones siguen las líneas que de forma natural se forman en la piel del pene y escroto se obtiene como resultado un colgajo óptimo para realizar una neouretra y recubrirla con piel, evitando incluso los injertos de mucosa. Estas modificaciones permiten que cualquier tipo de hipospadias sea reparado en un único tiempo con muy buenos resultados a largo plazo.

Hipospadias proximales reparados en un único tiempo. Lecciones aprendidas tras 196 intervenciones primarias / Single-repair for primary proximal hypospadias. Lessons learned after 196 primary repairs

Introducción y objetivo. Los hipospadias peneanos proximales, escrotales y perineales son un reto para el cirujano. Tras toda una carrera dedicada a los hipospadias, la autora principal del trabajo resume su experiencia y demuestra que prácticamente cualquier tipo de hipospadias puede ser resuelto completamente en un único tiempo. Método. Se trata de una revisión retrospectiva y la discusión de una serie de hipospadias proximales consecutivos tratados en una única intervención por el mismo equipo quirúrgico entre 1999 y 2016. Resultados. 196 pacientes fueron intervenidos. El 68% de los casos quedaron satisfactoriamente resueltos tras la operación. Todos los hipospadias fueron reparados con una de las siguientes técnicas: onlay, colgajo prepucial tubularizado (Duckett) o injertos de mucosa. Los injertos se utilizaron solo hasta 2005; posteriormente, el refinamiento de las técnicas quirúrgicas expuestas permitió que cualquier hipospadias fuera reparado con las técnicas de onlay o Duckett. Solo un 13% de los casos necesitó algún tipo de procedimiento quirúrgico adicional, más allá del cierre de una fístula uretral puntiforme (presente en el 25% de los pacientes).Conclusión. El diseño de las incisiones cutáneas es fundamental en la reparación de los hipospadias. Cuando las incisiones siguen las líneas que de forma natural se forman en la piel del pene y escroto se obtiene como resultado un colgajo óptimo para realizar una neouretra y recubrirla con piel, evitando incluso los injertos de mucosa. Estas modificaciones permiten que cualquier tipo de hipospadias sea reparado en un único tiempo con muy buenos resultados a largo plazo

Purpose. Proximal, scrotal and perineal hypospadias challenge the surgeon. After 40 years devoted to hypospadias surgery, the senior author summarizes her experience, share her tricks and shows that almost any hypospadias can be fully repaired in one surgery. Methods. Retrospective review and discussions of a large series of consecutive proximal hypospadias treated in one surgery by the same team from 1999 to 2016. Results. 196 patients were operated. 68% of all patients were successfully repaired after the surgery. All hypospadias were treated using one of the following techniques: onlay double flap, tubularized preputial flap or mucosal grafts (only until 2005). Urethrocutaneous fistula was present in 25% of cases, but only 13% of patients needed other further surgical procedures. Conclusion. Cutaneous incisions design is fundamental in hypo-spadias repair. When the incisions follow the natural folds that appear in penile and scrotal skin, it's possible to obtain optimal flaps for both urethroplasty and skin coverage (even avoiding the need for mucosal grafts). Consequently, almost all kinds of hypospadias can be repaired in one surgery with very good long-term results

Topical estradiol increases epidermal thickness and dermal collagen of foreskin prior to hypospadia surgery - Randomized double blinded controlled trial.

INTRODUCTION: The use of preoperative topical testosterone stimulation prior to hypospadias correction aims to increase penile size and achieve better surgical results. Topical estradiol has been shown to improve the quality of skin in other sites, but its use in boys with hypospadia has not yet been elucidated. OBJECTIVE: This study aims to evaluate the primary effects in epidermal thickness and collagen distribution of estradiol compared to testosterone and placebo in skin of prepuce before hypospadia surgery. MATHERIALS AND METHODS: Patients were randomized into three groups according to the topical hormone used: TG: Testosterone ointment; EG: Estradiol ointment; CG: Neutral base ointment. Fragments of foreskin were excised, fixed and then sectioned for histology. For each sample, epidermal thickness and dermal collagen expression was measured by specific computer analysis, P-values of <0.05. RESULTS: Thirty-three patients with a mean age of 4.01 ± 2.92 years were included. Hypospadia classification was similar in all three groups. Mean epidermal thickness and collagen type I expression in EG were greater than those of the other groups. Collagen type III expression was similar in all groups. DISCUSSION: Foreskin has a fundamental role in many techniques of hypospadias surgery and can be used either as a graft or a flap in the correction of the penile defect. Increase of epidermal thickness and dermal collagen observed in the present study has already been related to use of estradiol in other skin sites, but not yet in foreskin. Further studies are needed to evaluate the real significance of these findings in boys with hypospadias. CONCLUSION: Use of topical estradiol before hypospadias surgery lead to greater epidermal thickness and increases dermal collagen expression in foreskin.

Selection of Anti-gluten DNA Aptamers in a Deep Eutectic Solvent.

Herein, we show the feasibility of using deep eutectic solvents as a faster way of selecting aptamers targeting poorly water-soluble species. This unexplored concept is illustrated for gluten proteins. In this way, aptamer-based gluten detection can be performed directly in the extraction media with improved detectability. We envision deep implications for applications not only in food safety control but also in biomedicine.

[When should gestation of a gastroschisis be terminated?] / ¿Cuándo debe terminarse la gestación de una gastrosquisis?

OBJECTIVES: In gastroschisis pregnancies, a) to correlate prenatal ultrasound variables with postnatal outcome and b) to determine the ideal time for setting the delivery in order to achieve the best neonatal outcome. MATERIAL AND METHODS: Retrospective review (2000-2015) of all available gastroschisis whose prenatal findings could be correlated with the neonatal outcome. Two study groups have been defined according to the complications present after birth: favorable gastrosquisis and complicated. Prenatal variables were compared by groups using McWhitney or Chi tests as needed. RESULTS: Twenty-two gastroschisis fulfilled the requirement. Twelve cases had uneventful outcomes. Ten patients experienced complications, including death in five. In the complicated group there were 15 episodes of sepsis and 17 reoperations. Any single ultrasound parameter could predict a bad follow up. In thirteen cases, delivery was forced due to sudden changes on ultrasound bowel appearance. Nine of these patients had very good neonatal outcome. CONCLUSIONS: Finishing pregnancy when sudden changes on the fetal bowel were identified was the only strategy that leaded us to diminish the complication rate in gastroschisis.

OBJETIVOS: En las gestaciones con gastrosquisis, a) valorar la presencia de algún dato ecográfico prenatal que pueda predecir la evolución postnatal de la gastrosquisis, y b) determinar el momento ideal del nacimiento de los pacientes con gastrosquisis que se relacione con una mejor evolución postnatal. MATERIAL Y METODOS: Revisión retrospectiva (2000-2015) de las gastrosquisis cuyos datos ecográficos prenatales hemos podido relacionar con las características de los pacientes y su evolución clínica posterior. Se han determinado dos grupos en función de la evolución favorable o complicada de la gastrosquisis. Todas las variables ecográficas prenatales se han comparado entre grupos según los test de McWitney o Chi cuadrado. RESULTADOS: Veintidós gastrosquisis cumplieron el requisito anterior. Doce casos tuvieron una evolución sin incidencias significativas. Diez pacientes tuvieron una evolución complicada, de los cuales cinco fueron exitus. En este grupo hubo 15 episodios de sepsis y 17 reintervenciones. Ningún parámetro ecográfico prenatal predijo con fiabilidad una evolución desfavorable. En 13 casos se finalizó la gestación porque aparecieron cambios súbitos en la ecografía. Nueve de estos pacientes evolucionaron sin ninguna complicación. CONCLUSIONES: Terminar la gestación cuando se produce un cambio súbito de la apariencia ecográfica de los intestinos fetales es la única estrategia que nos ha permitido disminuir la incidencia de complicaciones en los pacientes con gastrosquisis.

Solid-phase helicase dependent amplification and electrochemical detection of Salmonella on highly stable oligonucleotide-modified ITO electrodes.

An on-surface isothermal helicase-dependent amplification is devised for simple, point-of-need quantification of bacterial genomes. The method relies on the enzyme-extension of a thiol-modified reverse primer anchored to indium tin oxide electrodes, which shows strikingly high thermal and storage stability. Amplification and electrochemical detection of only 10 genomes are thus performed on the same platform without thermal cycling.

¿Cuándo debe terminarse la gestación de una gastrosquisis? / When should gestation of a gastroschisis be terminated?

Objetivos. En las gestaciones con gastrosquisis, a) valorar la presencia de algún dato ecográfico prenatal que pueda predecir la evolución postnatal de la gastrosquisis, y b) determinar el momento ideal del nacimiento de los pacientes con gastrosquisis que se relacione con una mejor evolución postnatal. Material y métodos. Revisión retrospectiva (2000-2015) de las gastrosquisis cuyos datos ecográficos prenatales hemos podido relacionar con las características de los pacientes y su evolución clínica posterior. Se han determinado dos grupos en función de la evolución favorable o complicada de la gastrosquisis. Todas las variables ecográficas prenatales se han comparado entre grupos según los test de McWitney o Chi cuadrado. Resultados. Veintidós gastrosquisis cumplieron el requisito anterior. Doce casos tuvieron una evolución sin incidencias significativas. Diez pacientes tuvieron una evolución complicada, de los cuales cinco fueron exitus. En este grupo hubo 15 episodios de sepsis y 17 reintervenciones. Ningún parámetro ecográfico prenatal predijo con fiabilidad una evolución desfavorable. En 13 casos se finalizó la gestación porque aparecieron cambios súbitos en la ecografía. Nueve de estos pacientes evolucionaron sin ninguna complicación. Conclusiones. Terminar la gestación cuando se produce un cambio súbito de la apariencia ecográfica de los intestinos fetales es la única estrategia que nos ha permitido disminuir la incidencia de complicaciones en los pacientes con gastrosquisis (AU)

Objectives. In gastroschisis pregnancies, a) to correlate prenatal ultrasound variables with postnatal outcome and b) to determine the ideal time for setting the delivery in order to achieve the best neonatal outcome. Material and methods. Retrospective review (2000-2015) of all available gastroschisis whose prenatal findings could be correlated with the neonatal outcome. Two study groups have been defined according to the complications present after birth: favorable gastrosquisis and complicated. Prenatal variables were compared by groups using McWhitney or Chi tests as needed. Results. Twenty-two gastroschisis fulfilled the requirement. Twelve cases had uneventful outcomes. Ten patients experienced complications, including death in five. In the complicated group there were 15 episodes of sepsis and 17 reoperations. Any single ultrasound parameter could predict a bad follow up. In thirteen cases, delivery was forced due to sudden changes on ultrasound bowel appearance. Nine of these patients had very good neonatal outcome. Conclusions. Finishing pregnancy when sudden changes on the fetal bowel were identified was the only strategy that leaded us to diminish the complication rate in gastroschisis (AU)

Electrochemical detection of magnetically-entrapped DNA sequences from complex samples by multiplexed enzymatic labelling: Application to a transgenic food/feed quantitative survey.

Monitoring of genetically modified organisms in food and feed demands molecular techniques that deliver accurate quantitative results. Electrochemical DNA detection has been widely described in this field, yet most reports convey qualitative data and application in processed food and feed samples is limited. Herein, the applicability of an electrochemical multiplex assay for DNA quantification in complex samples is assessed. The method consists of the simultaneous magnetic entrapment via sandwich hybridisation of two DNA sequences (event-specific and taxon-specific) onto the surface of magnetic microparticles, followed by bienzymatic labelling. As proof-of-concept, we report its application in a transgenic food/feed survey where relative quantification (two-target approach) of Roundup Ready Soybean® (RRS) was performed in food and feed. Quantitative coupling to end-point PCR was performed and calibration was achieved from 22 and 243 DNA copies spanning two orders of magnitude for the event and taxon-specific sequences, respectively. We collected a total of 33 soybean-containing samples acquired in local supermarkets, four out of which were found to contain undeclared presence of genetically modified soybean. A real-time PCR method was used to verify these findings. High correlation was found between results, indicating the suitability of the proposed multiplex method for food and feed monitoring.

Intraepithelial lymphocytes subsets in different forms of celiac disease.

AIM: The enumeration of intraepithelial lymphocytes subsets (total, γδ, and CD3(-) IELs) by flow cytometry (FCM), named as IEL lymphogram, constitutes a useful tool for celiac disease (CD) diagnosis. The aim of this study was to quantify IELs by FCM and their diagnostic value to differentiate active, silent and potential CD. METHODS: Prospective study of 60 active and 20 silent CD patients, and 161 controls in which duodenal biopsy and IEL quantification by FCM was performed. RESULTS: Active and silent CD patients had significant higher levels of both total and γδ IELs than absent CD patients (P < 0.0001 and P < 0.0001, P = 0.012 and P < 0.011; respectively). Active and silent CD patients had significant lower levels of CD3(-) IELs than absent CD patients (P < 0.047 and P < 0.009, respectively). Moreover, they were lower in silent than in active CD patients (P = 0.002). Changes of IELs subsets were more marked in children than adults active CD. The optimal IEL lymphogram cut off values for active CD diagnosis were: ≥10, ≥15 and ≤9 %, and with better performance characteristics for silent CD: ≥ 11, ≥10 and ≤5 %. CONCLUSION: The evaluation of IELs subsets by FCM is useful to confirm diagnosis of active and silent CD.

Comparison of isothermal helicase-dependent amplification and PCR for the detection of Mycobacterium tuberculosis by an electrochemical genomagnetic assay.

Methods for the early and sensitive detection of pathogenic bacteria suited to low-resource settings could impact diagnosis and management of diseases. Helicase-dependent isothermal amplification (HDA) is an ideal tool for this purpose, especially when combined with a sequence-specific detection method able to improve the selectivity of the assay. The implementation of this approach requires that its analytical performance is shown to be comparable with the gold standard method, polymerase chain reaction (PCR). In this study, we optimize and compare the asymmetric amplification of an 84-base-long DNA sequence specific for Mycobacterium tuberculosis by PCR and HDA, using an electrochemical genomagnetic assay for hybridization-based detection of the obtained single-stranded amplicons. The results indicate the generalizability of the magnetic platform with electrochemical detection for quantifying amplification products without previous purification. Moreover, we demonstrate that under optimal conditions the same gene can be amplified by either PCR or HDA, allowing the detection of as low as 30 copies of the target gene sequence with acceptable reproducibility. Both assays have been applied to the detection of M. tuberculosis in sputum, urine, and pleural fluid samples with comparable results. Simplicity and isothermal nature of HDA offer great potential for the development of point-of-care devices. Graphical Abstract Comparative evaluation of isothermal helicase-dependent amplification and PCR for electrochemical detection of Mycobacterium tuberculosis.

Estilo de vida y estado de nutrición en niños escolares / Lifestyle and nutritional status among school children / Estilo de vida e estado de nutrição em crianças na escola

Objetivo: Relacionar el estilo de vida y el estado de nutrición en escolares. Método: Estudio correlacional, participaron 260 niños adscritos a la Clínica ISSSTE, elegidos aleatoriamente. Se utilizó el cuestionario sobre estilos de vida en la niñez y se valoró el peso y talla. Se obtuvo la aprobación de la Jefatura de Investigación del ISSSTE. En el análisis de datos se aplicó estadística descriptiva e inferencial. Resultados: Participaron 137 niños y 123 niñas, con edades entre 6-12 años. Respecto al estado de nutrición, 5 de cada 10 de los participantes se encuentran en peso normal, 2 de cada 10 en sobrepeso y uno de cada 10 en obesidad y peso bajo. Sobre el estilo de vida 7 de cada 10 de los participantes se encontraron en riesgo bajo y 2 de cada 10 en riesgo mediano. Por otra parte, no se encontró relación entre el estilo de vida de los niños y su estado de nutrición (r s = 0.038, p > 0.05). Adicionalmente, se encontró que los niños con peso normal o bajo peso tienen mejores hábitos de sueño que los niños con sobrepeso u obesidad (U = 3.79, p < 0.001). Conclusiones: Este estudio muestra indicios de que los aspectos vinculados con la dimensión psicocorporal y con uso del tiempo libre deberían indagarse a mayor profundidad, ya que fueron los que arrojaron cifras que ubican a los escolares en riesgo medio y alto.

Objective: To explore the association between lifestyle and nutritional status among school children. Method: Correlational study involving 260 children registered at an ISSSTE clinic, who were randomly selected. A questionnaire on childhood lifestyles was used; weight and height were also assessed. A project approval from the ISSSTE research committee was obtained. Descriptive and inferential statistics were used. Results: 137 boys and 123 girls with ages between 6 and 12 years old participated. In relation to the nutrition status, five out every ten have a normal weight; two out every ten are over weighted, and one out of every ten is obese or under weighted respectively. In relation to lifestyle, seven out of every ten participants have low risk; and two out of every ten have medium risk. On the other hand a relation between the lifestyle and nutritional status of the children was not found (rs=.038, p > 0.05). Moreover, it was found that children with normal or low weights have better sleep habits in comparison to the children who are over weighted or obese U = 3.79, p < .001). Conclusions: This study suggests that issues related to the psychological and corporal dimensions, as well as the use of free time, should be further explored because they showed figures which place children within mid and high risks.

Objetivo: Relacionar o estilo de vida e o estado de nutrição das crianças na escola. Método: Estudo de correlação, em que participaram 260 crianças vinculadas à clínica ISSSTE, escolhidas aleatoriamente. Utilizou-se um questionário sobre estilos de vida na infância e valorizou-se o peso e o tamanho. Obteve-se a aprovação da chefia de pesquisa do ISSSTE. Na análise de dados aplicou-se estatística descritiva e inferencial. Resultados: Participaram 137 meninos e 123 meninas, com idade entre 6-12 anos de idade. Em relação ao estado de nutrição, 5 de cada 10 dos participantes encontram-se em peso normal, 2 de cada 10 com sobrepeso e um de cada 10 com obesidade e baixo peso. Sobre o estilo de vida, 7 de cada 10 dos participantes encontraram-se em risco de baixo peso e 2 de cada 10 em risco moderado. Por outra parte, não se encontrou relação entre o estilo de vida das crianças e o seu estado de nutrição (r s = 0.038, p > 0.05). Adicionalmente, encontrou-se que as crianças com peso normal ou baixo peso têm melhores hábitos de sono do que as crianças com sobrepeso ou obesidade (U = 3.79, p < 0.001). Conclusões: Esse estudo mostra indícios de que os aspetos vinculados com a dimensão psicocorporal e com uso do tempo livre deveriam ser indagados com maior profundidade, já que foram os que lançaram dados que situam as crianças na escola em risco moderado e alto.

Fetoscopic laser surgery to decompress distal urethral obstruction caused by prolapsed ureterocele.

We report on the successful use of fetoscopic surgery to treat a case of prolapsed ureterocele in a female fetus. At 21 weeks' gestation, a double renal system with an intravesical ureterocele obstructing the bladder outlet was diagnosed, causing severe megacystis, bilateral hydronephrosis and progressive oligohydramnios. Ultrasound evaluation following referral to our center confirmed severe bilateral hydronephrosis with pelvic and calyceal dilatation, but amniotic fluid volume was normal and the ureterocele was not visualized in the bladder. Instead, a cystic mass within the external genitalia was observed, suggestive of a prolapsed ureterocele, causing intermittently severe obstruction of the urethra. The parents were counseled about the uncertain prognosis and fetal surgery to decompress the urinary system was proposed. The procedure involved firing a contact diode laser until perforation of the ureterocele was achieved. Following laser surgery, resolution of megacystis, reduction of hydronephrosis and normalization of amniotic fluid volume were observed. Our report demonstrates that fetoscopic decompression of a distal urethral obstruction is feasible in the rare event of congenital prolapsed ureterocele.

Development of a genosensor for peanut allergen ARA h 2 detection and its optimization by surface response methodology.

A new selective electrochemical genosensor has been developed for the detection of an 86-mer DNA peanut sequence encoding part of the allergen Ara h 2 (conglutin-homolog protein). The method is based on a sandwich format, which presents two advantages: it permits shortening the capture probe and avoids labeling of the target. Screen-printed gold electrodes have been used as platform for the immobilization of oligonucleotides by the well-known S-Au bond. Mixed self-assembled monolayers (SAM), including thiol-modified capture probe and mercaptohexanol, were prepared to achieve an organized, homogeneous and not too compact SAM in which unspecific adsorption of the capture probe would be prevented. The optimization of the sensing phase was carried out using the Design of Experiments (DoE) approach. Traditionally, response optimization is achieved by changing the value of one factor at a time until there is no further improvement. However, DoE involves regulating the important factors so that the result becomes optimal. Optimized conditions were found to be 1.34 µM for capture probe concentration and 3.15 mM for mercaptohexanol (spacer) concentration. When the optimal conditions were employed the analytical performance of the proposed genosensor improved significantly, showing a sensitivity as high as 3 µA/nM, with a linear range from 5×10(-11) to 5×10(-8) M and a detection limit of 10 pM.

Detección de alérgenos de cacahuete mediante un sensor de ADN / Detection of peanut allergens by a DNA sensor

En el presente trabajo se propone un genosensor electroquímico para la detección de un segmento de ADN que codifica parte de la proteína alergénica Ara h 2 del cacahuete. El genosensor se basa en un ensayo tipo sándwich, el analito hibrida con dos secuencias de bases, una de ellas inmovilizada sobre un electrodo de oro serigrafiado, formando una monocapa autoensamblada. La optimización del dispositivo se realizó utilizando la metodología de Superficies de Respuesta. La máxima respuesta se encontró para concentraciones de sonda de captura y agente bloqueante, 1 mM y 2,5 mM respectivamente

In the present work an electrochemical genosensor for detecting a DNA segment encoding part of the allergenic protein peanut Ara h 2 is proposed. Genosensors is based on a sandwich assay format, the analyte hybridized with two base sequences, one immobilized onto a screen printed gold electrode, forming a self-assembled monolayer. The optimization of the device was performed using Response Surface Methodology. The maximum response was found to be 1 µM of capture probe concentration and 2,5 mM of blocking agent concentration

Flujo mesentérico en un modelo experimental por isquemia-reperfusión en ratas / Mesenteric flow in an experimental model of ischaemia-reperfusion in rats

INTRODUCCIÓN: La oclusión aguda mantenida seuida de la reperfusión de la arteria mesentérica superior (AMS) puede desencadenar en pocas horas el daño irreversible del intestino. Nuestro objetivo fue determinar los cambios de flujo mesentérico medidos por ecografía Doppler color y la correlación con las lesiones histológicas en un modelo experimental de isquemia-reperfusión. MATERIAL Y MÉTODO: Se estudiaron 3 grupos (control, isquemia y reperfusión) de ratas Sprague-Dawley de 17 días de vida. El modelo utilizado fue de isquemia-reperfusión sobre la AMS. Posteriormente, realizamos una ecografía intraabdominal. Las variables ecográficas fueron: velocidad sistólica máxima (VSM), velocidad media (Vm), flujo diastólico (FD) y los índices de pulsatilidad (IP), resistencia (IR) y sístole/diástole (S/D). Las variables histológicas fueron: lesión intestinal (escala Wallace/Keenan y Chiu), morfométricas (altura [AMV] y espesor medio de vellosidades) y células caliciformes. Utilizamos la correlación de Sperman (rs). RESULTADOS: En el grupo reperfusión la VSM (74,3 cm/s), el IP (7,33) y S/D (25,75) en la AMS se encuentran aumentados respecto a los controles (41,35 cm/s [3,12]; [12.45]). La VSM, IP y S/D establecieron regresiones significativas (p < 0,01) con: Wallace/Keenan (rs = 0,655; rs = 0,593; rs = 0,63), Chiu delgado (rs = 0,569; rs = 0,522; rs = 0,47), la AMV (rs = -0,495; rs = -0,452; rs = -0,459), y células caliciformes del colon (rs = -0,525; rs = -0,45; rs = -0,518). CONCLUSIONES: En la fase de reperfusión el incremento del flujo mesentérico expresado por la VSM y el IP podría pronosticar de forma significativa el potencial daño intestinal que ocurre a nivel macroscópico y microscópico

INTRODUCTION: Maintained acute occlusion followed by reperfusion of the superior mesenteric artery (SMA) in a few hours can trigger irreversible bowel damage. The aim of the study was to determine the changes in mesenteric flow measured by colour Doppler Ultrasound and correlating with histological lesions in an experimental model of ischaemia-reperfusion. Method and material: Three groups of Sprague-Dawley 17 day-old rats were studied (control, ischemia and reperfusion). The model used was ischaemia-reperfusion over the SMA. Intraabdominal ultrasound was then performed. The parameters recorded were: Maximum systolic velocity (MSV), pulsatility index (PI), resistance (RI) and systole-diastole (S/D). The histological variables were: intestinal lesion (Wallace/Keenan-Chiu scale), morphometrics (mean villus height [MVH]), and goblet cells. The Spearman (rs) correlation was used. RESULTS: The MSV in the reperfusion group was 74.3 cm/s, the PI 7.33 and S/D 25.75 in the SMA, which were higher than the controls (41.35 cm/s; 3.12 and 12.45, respectively). A direct association (P<.01) was found between MSV, PI and S/D regarding: Wallace/Kennan scoring system (rs = 0.655; rs = 0.593; rs = 0.63) and the Chiu (rs = 0.569; rs = 0.522; rs = 0.47). While the correlation was the reverse (P<0.01) when associated with the MVH (rs = -0,495; rs = -0,452;rs = -0,459) and goblet cells of the colon (rs = -0,525; rs = -0,45; rs = -0,518).CONCLUSIONS: The reperfusion phase increased mesenteric flow expressed by the MSV and PI and could significantly predict the potential bowel damage at macroscopic and microscopic level

Aptamer binding to celiac disease-triggering hydrophobic proteins: a sensitive gluten detection approach.

Celiac disease represents a significant public health problem in large parts of the world. A major hurdle in the effective management of the disease by celiac sufferers is the sensitivity of the current available methods for assessing gluten contents in food. In response, we report a highly sensitive approach for gluten analysis using aptamers as specific receptors. Gliadins, a fraction of gluten proteins, are the main constituent responsible for triggering the disease. However, they are highly hydrophobic and large molecules, regarded as difficult targets for in vitro evolution of aptamers without nucleobase modification. We describe the successful selection of aptamers for these water insoluble prolamins that was achieved choosing the immunodominant apolar peptide from α2-gliadin as a target for selection. All aptamers evolved are able to bind the target in its native environment within the natural protein. The best nonprotein receptor is the basis for an electrochemical competitive enzyme-linked assay on magnetic particles, which allows the measurement of as low as 0.5 ppb of gliadin standard (0.5 ppm of gluten). Reference immunoassay for detecting the same target has a limit of detection of 3 ppm, 6 times less sensitive than this method. Importantly, it also displays high specificity, detecting the other three prolamins toxic for celiac patients and not showing cross-reactivity to nontoxic proteins such as maize, soya, and rice. These features make the proposed method a valuable tool for gluten detection in foods.