Analysis of hypervariable DNA sequences by NGS technologies: QuasiFlow

The development of Next Generation Sequencing (NGS) technologies has allowed deep characterization of highly variable sequences such as viral or mitochondrial genomes. With respect to RNA and ssDNA viruses, their low replication fidelity generates viral populations consisting of complex mutant spectra termed viral quasispecies. Their study is of special interest as they can be considered a phenotypic reservoir1. Similarly, heteroplasmy of human mitochondrial genomes, in which different sequences are found within a single individual, might have important clinical consequences. For the analysis of the mutant spectrum of such hypervariable sequences from NGS data, we have developed QuasiFlow, a workflow designed in AutoFlow2 that uses Illumina reads. QuasiFlow provides information about DNA variability, such as SNPs, indels and recombination events (Figure 1). Furthermore, it allows haplotype reconstruction of viral quasispecies and characterization of its diversity through normalized Shannon index, nucleotide diversity and mutation networks. Quasiflow performs also a comparative study among samples, based on correlation, ANOVA and PCA analysis, in order to determine which parameters are affected by the experiment and how the samples behave according to their biological origin. In this work, we have applied QuasiFlow to analyze the population structure of the begomovirus Tomato yellow leaf curl virus (TYLCV) infectious clone inoculated in Arabidopsis thaliana plants, using HiSeq or MiSeq reads. Their analysis allowed detection of minor quasispecies variants with a frequency of 10-4 to 10-5 and reconstructed the haplotypes present in the sample. In addition, QuasiFlow was used to discover variants and recombinants in mixed infections of tomato plants. These results show the fast generation of recombinant genomes in geminivirus mixed infections and demonstrate the potential of QuasiFlow for the analysis of mutant spectra using Illumina MiSeq sequencing data. We have extended the use of QuasiFlow to the analysis of highly variable sequences such as the mitochondrial DNA. For that, we have analyzed DNA Illumina Miseq reads from 47 human mitochondrial samples from different cell lines obtained from the NCBI SRA database. Quasiflow generated automatically SNPs, SNP frequencies, indels and analyzed up to 23 variables using PCA analysis and performed an hierarchical clustering of the samples. Our analysis was able to detect pathological variants presented in a frequency lower than 1%.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. This research was funded by Junta de Andalucía and EU through the ERDF 2014-2020, Projects P10-CVI-6075 to M. G.C. and P10-CVI-6561 to A.G-P

Síndrome compartimental crónico de esfuerzo Afectación bilateral del compartimento anteroexterno de la pierna en deportistas

Se presenta nuestra experiencia en el manejo de (i casos de síndrome compartimental crónico del esfuerzo por afectación colateral del compartimento anteroexterno del EEII. El diagnóstico se confirmó mediante medición de presiones intracompartimentales con catéter de hendidura, considerando patológico cuando ante clínica compatible la presión basal no se recupera en al menos 15 minutos. Se realizó doble fasciotomía subcutánea en los 6 casos consiguiendo la desaparición de la sintomatología y reincorporación a los entrenamientos en 6 semanas en todos los casos. No se han observado complicaciones ni recidivas tras un seguimiento medio de 2 años (1-3).Our experience on the management of 6 cases with bilateral chronic exertional compartment syndrome of the legs is presented. The diagnosis was confirmed by the measurement of the compartment pressure with slit catheter. Intramuscular pressure was considered abnormal when the basal pressure was not regained 15 minutes after the exercise with consistent symtomatology. Double complete subcutaneous fasciotomy was performed in the 6 patients obtaining complete recovery of the symtoms and improvement of the performance in all of them. Patients started training six weeks after surgery. Neither complication or relapse was found during a mean follow-up of 2 years (1-3)

Fractura-avulsión de la tuberosidad tibial anterior en adolescentes: a propósito de dos casos. Revisión de la literatura

La avulsión de la tuberosidad tibial anterior es una lesión rara que se caracteriza por ocurrir en adolescentes deportistas. Esta lesión ocurre en un período vulnerable de la adolescencia, cuando la fisis sufre cambios fisiológicos que debilitan su resistencia a las fuerzas de tensión. Aportamos dos nuevos casos que fueron tratados con dos métodos distintos de osteosíntesis (tornillos y agujas de Kirschner) y realizamos una completa revisión bibliográfica. La osteosíntesis proporciona una mayor seguridad y menor tiempo de inmovilización, siendo el resultado final la epifisiodesis de dicha tuberosidad. Reseñamos la aparición de calcificaciones asintomáticas en el tendón rotuliano, complicación hasta ahora no descrita.Avulsion of the tibial tubercle is a rare lesion ocurring in sportif adolescents. This injury occurs in a vulnerable period that happen in adolescence when the physis is undergoing phisiologic change s that weaken its resistenc e to tensil loading. We present two ne w case s treated by two different procedures (Kirschner wire s and screws) and we made a complete literature review. We emphasize the finding of asintomatic calcification into patellar tendon that have not been previously described

Comparación de Resultados del electromiograma y de la saculorradiculografía en el Diagnóstico de las Hernias Discales

Los autores presentan un estudio comparativo de E.M.G. y Saculorradiculografía en relación con los hallazgos quirúrgicos de 100 Hernias Discales intervenidas. Para el E.M.G. se ha conseguido una exactitud global del 83% y para la saculorradiculografía del 79%. La saculorradiculografía ha demostrado mayor exactitud para la raiz S1 que paral a L5 (85,7% frente a 70,4%) y el E.M.G. mayor para la L5 que para la S1 (88,6% frente a 78,5%). El E.M.G. tiene una especial utilidad para el diagnóstico y despistaje de neuropatías y para la comprobación de recuperación post-quirúrgica de lesiones sin axonotmesis. La exactitud de los resultados en el E.M.G. están en relación directa con la habilidad del examinador a la hora de conseguir e interpretar los resultados.The authors made a comparative study between the Electromyography (EMG) and the radiculography with the surgical findings in 100 Herniated intervertebral disk (H.I.D.) operated on. With the E.M.G. they had obtained a correct diagnostic in 83% of case s and 79% with the radiculography. The radiculography showed the bigger accurancy in S1 than L5 (85'7% - 70'4%) and E.M.G. showed bigger accurancy B in L5 than S1 (88'6% - 78'5%). The E.M.G. had an special utility in the diagnosis of neuropathie s and the review and follow up after surgical procedure. It's very important the trainning of the E.M.G. examine r to obtain correct information

Resultados a largo plazo de intervención de Magnuson-Stack modificada para la luxación anterior recidivante de hombro

Los autores han realizado una revisión largo plazo versus corto plazo con 29 pacientes tratados mediante la técnica de Magnuson-Stack por luxación anterior recidivante de hombro. Se encontró un empobrecimiento de los resultados globales, observándose empeoramiento de la estabilidad (pasan del 90.47% de resultados excelentes al 58.62%, así como del 9.52 de resultados malos al 31.03%), aumento del arco de movilidad (movilidad excelente del 57.14 al 82.75%), permaneciendo la función prácticamente inalterada.Twenty-nine patients with recurrent shoulder dislocation trailed surgicaly by modified Magnuson-Stack procedure are colected. The authors made a long-term versus short-term review of results. They have found a worsening of results in the total longterm review. They observe a worsening of the stability (90.47% excellent results in short-term but 58.62% in long-term) (9.52% bad results in short-term but 31.03% in long-term). There are an increase of range mobility (57.14% excellent in short-term versus 82.75% in long-term). The function remain unchanged

Analysis of viral quasispecies by NGS technologies: quasiflow

We have developed QuasiFlow, a workflow designed in AutoFlow that takes advantage of NGS technologies to reconstruct quasispecies based in Illumina reads. QuasiFlow characterises and computes several key parameters, such as recombination events, SNPs, transitions, transversions, indels, quasispecies reconstruction, normalized Shannon index, nucleotide diversity and mutation networks. Moreover, it performs a comparative study of the samples comprising correlation, ANOVA and PCA analyses of the previously obtained virus population parameters. Using QuasiFlow we have analysed Illumina MiSeq reads from DNA samples obtained in mixed infections of ssDNA begomovirus in tomato plants amplified by rolling circle amplification. Further, we have extended the use of QuasiFlow to the analysis of the highly variable mitochondrial DNA. For that, we have used DNA Illumina MiSeq reads from 47 human mitochondrial samples from different cell lines obtained from the NCBI SRA databaseUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Valoración a medio-largo plazo de los cotilos de Mittelmeier

El objetivo del presente trabajo es valorar clínica y radiológicamente la evolución de 125 cotilos de Mittelmeier implantados en nuestro servicio, con un seguimiento medio de 8 años (5,5-14). En toda la serie se ha utilizado un mismo tipo de cotilos, aunque se han utilizado 2 vástagos, los de segunda y los de tercera generación. El comportamiento ha sido satisfactorio en el 91% de los de tercera generación y el 57% en los de segunda. Se comprobó una inclinación y recubrimientos progresivos y una emigración horizontal y vertical significativas de los cotilos. No se encontró relación entre los resultados clínicos y las alteraciones posicionales. La movilización del cotilo aparece en nuestra serie mucho más frecuentemente que en la mayor parte de la literatura. Las emigraciones horizontal y vertical pueden ser debidas a los diferentes coeficientes de elasticidad de la cerámica y el hueso sin implicar necesariamente malos resultados. Como conclusión, el comportamiento de los cotilos de Mittelmeier ha sido en general satisfactorio a pesar de las emigraciones, que han tenido escasa repercusión clínica.This study was aimed at evaluating clinically and radiologically the results of 125 Mittelmeier cups implanted in our institution, with a mean follow-up of 8 years (5.5-14). The same type of cup was used in the whole series, combined with two types of femoral stem from the 2 nd and 3rd generations. The results were satisfactory in 91 % of cases treated with the 3rd generation and 57% in those cases treated with the 2nd generation of implants. Progressive inclination and coverage were detected with both horizontal and vertical migration of the cups. No relationship was found between clinical results and the displacements observed. In this series, migrations of the cup were more frequent than those published by other authors. Horizontal and vertical migratrion may be due to the difference between the coefficients of elasticity of ceramic and bone. Cup migration did not imply poor results. In conclusion, the overall results obtained with the Mittelmeier cups were satisfactory, in spite of migration. The clinical repercussion of cup migrations was minimal

Sacrolistesis postraumática sin lesión neurológica

Las fracturas transversas de sacro son raras pero aún lo son más aquellas en las que hay un desplazamiento o listeis de una vértebra sacra sobre otra. El sentido de desplazamiento viene condicionado por el mecanismo causante, pudiendo ser en flexión o en extensión. En las verdaderas listesis el sentido del traumatismo es en extensión, llevando a colocarse total o parcialmente la vértebra sacra superior por delante de la inferior. En determinados casos como pueden ser los politraumatizados, el diagnóstico puede ser difícil, por lo que es necesario realizar un estudio radiológico meticuloso. En estas fracturas ocurre frecuentemente una lesión de raíces lumbosacras de mayor o menor gravedad, aunque suele existir algún tipo de recuperación, incluso espontánea, con el transcurso del tiempo. Se presenta el caso de una mujer que presentó una sacrolistesis de S1 sobre S2 por haber sufrido caída de nalgas, sin existir ninguna lesión neurológica.Transverse fractures of the sacrum are rare, but fractures with displacement or listhesis of one sacral vertebra over another are even more infrequent. The direction of the displacement is conditioned by the causing mechanism and may occur in flexion or extension, in true listhesis, trauma occurs in extension, placing the superior sacral vertebra totally or partially in from of the inferior vertebra. In some cases, such as multiple traumas, the diagnosis can be difficult, and a careful radiologic study is required. A more or less severe injury of th lumbosacral roots frequently occurs in these fractures, although a certain degree of recovery takes place, even spontaneously, over time. We present the case of a 69 years old woman suffering from a sacrolisthesis of S1 over S2 after a fall on the buttocks with no neurological lesion

Comparison of Two Energy Management System Strategies for Real-Time Operation of Isolated Hybrid Microgrids

The propagation of hybrid power systems (solar–diesel–battery) has led to the development of new energy management system (EMS) strategies for the effective management of all power generation technologies related to hybrid microgrids. This paper proposes two novel EMS strategies for isolated hybrid microgrids, highlighting their strengths and weaknesses using simulations. The proposed strategies are different from the EMS strategies reported thus far in the literature because the former enable the real-time operation of the hybrid microgrid, which always guarantees the correct operation of a microgrid. The priority EMS strategy works by assigning a priority order, while the optimal EMS strategy is based on an optimization criterion, which is set as the minimum marginal cost in this case. The results have been obtained using MATLAB/Simulink to verify and compare the effectiveness of the proposed strategies, through a dynamic microgrid model to simulate the conditions of a real-time operation. The differences in the EMS strategies as well as their individual strengths and weaknesses, are presented and discussed. The results show that the proposed EMS strategies can manage the system operation under different scenarios and help power system operator obtain the optimal operation schemes of the microgrid.This work was supported by the Autonomous Community of Madrid under the PROMINT-CM project (S2018/EMT-4366)

Lipomatosis epidural lumbosacra idiopática: revisión de la bibliografía y caso clínico

La lipomatosis epidural es una afección que aparece generalmente en relación con hipercortisolismos endógenos o exógenos, siendo más raras las formas idiopáticas, de las que se han descrito únicamente 17 casos. Se presenta un nuevo caso con sintomatología compatible con estenosis de canal, y que tras estudios de imagen se demostró que correspondía con una lipomatosis epidural lumbosacra. Se le encomendó al paciente una reducción de peso, con lo que se alivió parcialmente la sintomatología, permaneciendo estable en la actualidad. Se revisa la bibliografía de este raro proceso, exponiendo los distintos hallazgos encontrados. Algunos autores proponen una reducción de peso, lo cual puede aliviar la sintomatología. Para casos rebeldes está indicada la laminectomía asociada al despegamiento de la grasa sin realizar exploración intradural.Epidural lipomatosis is an affection that is usually related to endogenic or exogenous hipercortisolism. Less frequently, it can be idiopathic. To date only 17 cases have been documented. A new case is presented in a patient suffering from symptoms resembling lumbar spinal stenosis. The radiological studies revealed a compression of the dural sac at lumbosacral levéis, caused by tissue similar in density to fat. Weight reduction, was recommended to the patients and a partial relief of symptoms was achieved. The authors reviewed the pathogeny of epidural lipomatosis and its various different treatments. In the idiopathic form, weight reduction is recomended first, reserving surgery for those cases that do not respond to this initial treatment. Surgical treatment should consist of laminectomy and fat debulking, and should not include intradural exploration