A High-Throughput Colocalization Pipeline for Quantification of Mitochondrial Targeting across Different Protein Types.

Efficient metabolic engineering and the development of mitochondrial therapeutics often rely upon the specific and strong import of foreign proteins into mitochondria. Fusing a protein to a mitochondria-bound signal peptide is a common method to localize proteins to mitochondria, but this strategy is not universally effective, with particular proteins empirically failing to localize. To help overcome this barrier, this work develops a generalizable and open-source framework to design proteins for mitochondrial import and quantify their specific localization. This Python-based pipeline quantitatively assesses the colocalization of different proteins previously used for precise genome editing in a high-throughput manner to reveal signal peptide-protein combinations that localize well in mitochondria.

High-throughput colocalization pipeline quantifies efficacy of mitochondrial targeting signals across different protein types.

Efficient metabolic engineering and the development of mitochondrial therapeutics often rely upon the specific and strong import of foreign proteins into mitochondria. Fusing a protein to a mitochondria-bound signal peptide is a common method to localize proteins to mitochondria, but this strategy is not universally effective with particular proteins empirically failing to localize. To help overcome this barrier, this work develops a generalizable and open-source framework to design proteins for mitochondrial import and quantify their specific localization. By using a Python-based pipeline to quantitatively assess the colocalization of different proteins previously used for precise genome editing in a high-throughput manner, we reveal signal peptide-protein combinations that localize well in mitochondria and, more broadly, general trends about the overall reliability of commonly used mitochondrial targeting signals.

Gene silencing in adult Popillia japonica through feeding of double-stranded RNA (dsRNA) complexed with branched amphiphilic peptide capsules (BAPCs).

Gene silencing by feeding double-stranded (dsRNA) holds promise as a novel pest management strategy. Nonetheless, degradation of dsRNA in the environment and within the insect gut, as well as inefficient systemic delivery are major limitations to applying this strategy. Branched amphiphilic peptide capsules (BAPCs) complexed with dsRNA have been used to successfully target genes outside and inside the gut epithelium upon ingestion. This suggests that BAPCs can protect dsRNA from degradation in the gut environment and successfully shuttle it across gut epithelium. In this study, our objectives were to 1) Determine whether feeding on BAPC-dsRNA complexes targeting a putative peritrophin gene of P. japonica would result in the suppression of gut peritrophin synthesis, and 2) gain insight into the cellular uptake mechanisms and transport of BAPC-dsRNA complexes across the larval midgut of P. japonica. Our results suggest that BAPC-dsRNA complexes are readily taken up by the midgut epithelium, and treatment of the tissue with endocytosis inhibitors effectively suppresses intracellular transport. Further, assessment of gene expression in BAPC- peritrophin dsRNA fed beetles demonstrated significant downregulation in mRNA levels relative to control and/or dsRNA alone. Our results demonstrated that BAPCs increase the efficacy of gene knockdown relative to dsRNA alone in P. japonica adults. To our knowledge, this is the first report on nanoparticle-mediated dsRNA delivery through feeding in P. japonica.

A Bioinformatic Algorithm based on Pulmonary Endoarterial Biopsy for Targeted Pulmonary Arterial Hypertension Therapy.

Background: Optimal pharmacological therapy for pulmonary arterial hypertension (PAH) remains unclear, as pathophysiological heterogeneity may affect therapeutic outcomes. A ranking methodology based on pulmonary vascular genetic expression analysis could assist in medication selection and potentially lead to improved prognosis. Objective: To describe a bioinformatics approach for ranking currently approved pulmonary arterial antihypertensive agents based on gene expression data derived from percutaneous endoarterial biopsies in an animal model of pulmonary hypertension. Methods: We created a chronic PAH model in Micro Yucatan female swine by surgical anastomosis of the left pulmonary artery to the descending aorta. A baseline catheterization, angiography and pulmonary endoarterial biopsy were performed. We obtained pulmonary vascular biopsy samples by passing a biopsy catheter through a long 8 French sheath, introduced via the carotid artery, into 2- to 3-mm peripheral pulmonary arteries. Serial procedures were performed on days 7, 21, 60, and 180 after surgical anastomosis. RNA microarray studies were performed on the biopsy samples. Results: Utilizing the medical literature, we developed a list of PAH therapeutic agents, along with a tabulation of genes affected by these agents. The effect on gene expression from pharmacogenomic interactions was used to rank PAH medications at each time point. The ranking process allowed the identification of a theoretical optimum three-medication regimen. Conclusion: We describe a new potential paradigm in the therapy for PAH, which would include endoarterial biopsy, molecular analysis and tailored pharmacological therapy for patients with PAH.

Response of Mexican life and non-life insurers to the low interest rate environment.

The purpose of this research is to examine whether insurers have improved their economic performance through efficiency as a reaction to the prolonged period of low interest rates. The results of an analysis of 22 years of data, using a two-stage data envelopment analysis approach (DEA), show that there is an inverse relationship between efficiency and interest rate. The non-life insurance group had a superior level of efficiency compared with that of the life insurance group. It seems that life insurance companies, to some extent, transferred their inefficiency to customers to maintain their solvency. Additionally, this research shows that companies with substantial market power and bancassurance exceeded the performance of the rest of the industry.

Metabolic Footprints of Burkholderia Sensu Lato Rhizosphere Bacteria Active against Maize Fusarium Pathogens.

Consistent with their reported abundance in soils, several Burkholderia sensu lato strains were isolated from the rhizosphere of maize plants cultivated at different sites in central México. Comparative analysis of their 16S rRNA gene sequences permitted their separation into three distinctive clades, which were further subdivided into six other clusters by their close resemblance to (1) Trinickia dinghuensis; (2) Paraburkholderia kirstenboschensis, P. graminis, P. dilworthii and P. rhynchosiae; (3) B. gladioli; (4) B. arboris; (5) B. contaminans, or (6) B. metallica representative species. Direct confrontation assays revealed that these strains inhibited the growth of pathogenic Fusarium oxysporum f. sp. radicis-lycopersici, and F. verticillioides within a roughly 3-55% inhibition range. The use of a DIESI-based non-targeted mass spectroscopy experimental strategy further indicated that this method is an option for rapid determination of the pathogen inhibitory capacity of Burkholderia sensu lato strains based solely on the analysis of their exometabolome. Furthermore, it showed that the highest anti-fungal activity observed in B. contaminans and B. arboris was associated with a distinctive abundance of certain m/z ions, some of which were identified as components of the ornbactin and pyochelin siderophores. These results highlight the chemical diversity of Burkholderia sensu lato bacteria and suggest that their capacity to inhibit the Fusarium-related infection of maize in suppressive soils is associated with siderophore synthesis.

Text segmentation ability predicts future reading efficiency in Spanish-speaking children.

Can the ability to parse unspaced texts (measured by a Text Segmentation Task, TST) index and predict reading efficiency in Spanish-speaking children? A sample of 1112 children (1st to 6th grade) was assessed. Additionally, two subsamples (51 children of 4th-5th grades and 71 children of 1st grade) were followed up. Our results indicate that the TST: a) reflects the acquisition of reading over primary school grades; b) reflects the teacher's judgment about the child's reading development; c) accurately predicts oral reading efficiency one and four years later year, in the former case even after removing the contributions of the IQ and oral reading speed. These results indicate that TST can be used to both index present -and predict future- reading achievements.

Measurement and analysis of broadband UVB solar radiation in Spain.

Measurements of broadband UVB irradiance (290-315 nm) at 14 locations in Spain for the period 2000-2009 have been used to generate instantaneous, hourly and daily values of irradiance (W m(-2)) and radiant exposure (kJ m(-2)). These measurements, and its statistical indices, have been analyzed. For the UVB irradiance, the values corresponding to July (maximum) and December (minimum) have been analyzed as representative of the year during the whole period for all locations. For the UVB radiant exposure, the temporal evolution of daily values has been evaluated for all locations to estimate an average yearly behavior. The accumulated radiant exposure for an average year has also been studied for each location. Finally, to determine possible trends in the evolution of the UVB levels, the linear regressions for the mean daily values for all locations have been determined.

Treatment of refractory epilepsy in adult patients with right-sided vagus nerve stimulation.

Vagus nerve stimulation (VNS) was introduced as a novel method for the treatment of patients with medically and/or surgically refractory epilepsy. VNS typically involves placement of the electrode on the left vagus nerve. However, there are some patients who cannot be implanted on this side because of complications related to the surgical procedure or side effects or infections forcing the left side VNS (L-VNS) explant. Because right side VNS (R-VNS) implants have shown similar effectiveness compared to L-VNS in reducing the frequency of seizures in animal models, treatment with R-VNS should be considered in patients who may not tolerate L-VNS. We present two adult patients who underwent R-VNS. One of the patients improved dramatically after L-VNS, but the device had to be removed because of mechanical malfunction. This patient was thought to be at high risk for nerve injury if L-VNS reimplantation was done, thus R-VNS was chosen. In the other patient, L-VNS was first attempted, but the operation had to be stopped due to significant bleeding caused by the accidental tearing of an ectopic vein. Both patients had a marked reduction in their seizure activity and none of them had cardiac side effects from therapeutic R-VNS. We conclude that R-VNS therapy is an alternative, promising therapy for reducing seizure activity in those patients who cannot undergo L-VNS implantation. Close follow-up and frequent ECG monitoring is required to detect the presence of cardiac side effects.

Seudoartrosis congénita de tibia. Reconstrucción con transplante de peroné vascularizado. Informe de 5 casos / Congenital pesuarthrosis of the tibia. reconstruction with fibular free flap. Report of five cases

Los tres principales retos que se plantean en la reconstrucción de una Seudoartrosis Congénita de Tibia (SCT) son: 1) lograr la unión ósea, 2) Corregir la discrepancia de longitud y 3) Minimizar la hiportrofia de la extremidad. Presentamos una serie de 5 pacientes con diagnóstico de SCT tipo II de Boyd, intervenidos entre los 18 meses y los 7 años meses (m;4a 2m) 4 pacientes con un total de 9 cirugías previas y uno solo sin antecedentes quirúrgicos. El acortamiento previo varió entre 3 y 7 cm. A todos los pacientes se les transfirió un peroné vascularizado con una longitud entre 8.7 y 14 cm, utilizando fijación externa en 3 y clavo centromedular en 2 casos. En todos los casos se obtuvo consolidación ósea: la unión proximal entre las 6 y 24 semanas y la distal entre las 6 y 16 semanas. En cuatro pacientes se retiró la osteosíntesis y se inició el apoyo gradual a partir de las 16 semanas. Se dio el índice de hipertrofia del peroné transplantado según de Boer y Wood, encontrando hipertrofia mayor del 20 por ciento a partir de la 4ª semana e hipertrofia endóstica entre la 8ª y 10ª semanas. Las complicaciones fueron: neuropraxia bilateral del sural externo que revirtió rápidamente (un caso), contractura del flexor hallucis longus manejada con servadoramante (un caso), infección del tracto de los clavos en un fijador que requirió de su retiro precozmente (un caso) y consolidación en valgo de tobillo que requirió de osteotomía correctora (un caso), El acortamiento residual al máximo seguimiento fue de entre 0 a 2.5 cm. La transferencia microvascular de peroné es el método más efectivo en el manejo actual de SCT tipo II de Boyd o multioperadas. Su morbilidad es menor que la reportada para otros métodos convencionales más reconocidos

Tratamiento de la esclerodid múltiple con interferón alfa-2B recombinante: estudio piloto

Se estudiaron 6 mujeres y 3 hombres (edad de 22 a 48 años) con esclerosis múltiple (EM) clínica definida (más de 2 años de evolución con al menos una recaída), lesiones características en imágenes de resonancia magnética, bandas oligoclonales de inmunoglobulinas en líquido cefalorraquídeo y sin alteraciones en las velocidades de conducción sensitiva y motora de las extremidades. Se administró 6 millones de UI de interferón alfa-2b recombinante (Heberón alfa R) intramuscular semanal durante 3 a 48 (promedio 24) meses. Seis pacientes tenían la forma evolutiva exacerbación-remisión (ER), 2 la exacerbación-progresiva(ER-SP) y una la exacerbación-remisión-estable (ER-ES). Después del tratamiento hubo menos recaídas de todo el grupo en comparación con las observadas en los 2 años anteriores. En el grupo ER, de 11 recaídas sólo se produjeron 3, de menor intensidad y duración. Tres de estos pacientes no tuvieron recaídas durante 24, 30 y 42 meses de tratamiento. Hubo mejoría de un punto o más en las evaluaciones de la escala de inhabilidad de Kurtzke en 3 pacientes con forma EM-ER y menor de un punto en otros 2. En todos los pacientes excepto una con la forma ER-ES hubo mejoría en la escala de evaluación neurológica Scripps. El tratamiento fue bien tolerado (AU)

Pseudoartrosis congénita de la tibia. Reconstrucción con transplante de peroné vascularizado. Informe de 5 casos / Congenital pseudoarthrosis of the tibia. Reconstruction by micro-vascular fibular graft. report o five cases

Presentamos una serie de 5 pacientes con diagnóstico de SCT tipo II de Boyd, intervenidos entre los 18 meses y los 7 años 2 meses (m: 4a 2m) 4 pacientes con un total de 9 cirugías previas y uno solo sin antecedentes quirúrgicos. El acortamiento previo varió entre 3 y 7 cm (m: 4.98 cm). A todos los pacientes se les transfirió un peroné vascularizado con una longitud entre 8.7 y 14 cm (m: 11.14 cm), utilizando fijación externa en 3 y clavo centromedular en 2 casos. En todos los casos se obtuvo consolidación ósea: la unión proximal entre las 6 y 24 semanas y la distal entre las 6 y 16 semanas. En cuatro pacientes se retiró la osteosíntesis y se inició el apoyo gradual a partir de las 16 semanas. Se midió el índice de hipertrofia del peroné transplantado según de Boer y Wood, encontrando hipertrofia mayor del 20 por ciento a partir de la cuarta semana e hipertrofia endóstica entre la octava y décima semanas. Las complicaciones fueron: neuropraxia bilateral del sural externo que revirtió rápidamente (un caso), contractura del flexor propio del dedo grueso manejada conservadoramente (un caso), infección del tracto de los clavos en un fijador que requirió de su retiro precozmente (un caso) y consolidación en valgo de tobillo que requirió de osteotomía correctora (un caso). El acortamiento residual al máximo seguimiento fue de entre 0 a 2.5 cm (m: 1.12 cm). La transferencia microvascular de peroné es el método más efectivo en el menejo actual de SCT tipo II de Boyd o multioperadas. Su morbilidad es menor que la reportada para otros métodos convencionales más reconocidos

Artroplastía por interposición del codo en el tratamiento de las secuelas traumáticas en el niño / Interpositión arthroplasty of the elbow for treatment in children whit traumatic sequelae

Las fracturas del miembro superior constituyen del 65 al 75 por ciento de estas lesiones en los niños. El área más afectada es el antebrazo seguida por el codo. En nuestro medio se observan con cierta frecuencia pacientes con lesiones traumáticas en el codo que no han sido tratadas o han recibido manejo empírico con las consecuencias secuelas en cuanto a limitación de la movilidad, inclusive con anquilosis en extensión. Se presentan los resultados del tratamiento quirúrgico con artroplastia anatomofuncional (McAusland) en 23 pacientes con un promedio de edad de 10.7 años y un tiempo de evolución preoperatoria de 12.4 meses en promedio, analizando también los factores que influyen en los mismos. Para la valoración de los resultados se tomaron en cuenta movilidad, dolor, estabilidad y función pre y postoperatorios obteniéndose buenos resultados en 22.7 por ciento de casos, regulares en 27.2 por ciento de pacientes, pobres en 31.8 por ciento y malos en 4 por ciento. La técnica anatomofuncional con interposición de fascia proporciona los mejores resultados en niños mayores de 10 años y con menos de seis meses de evolución es el problema principal no resuelto pero al menos se obtiene una articulación en posición funcional, estable, no dolorosa y grados variables de movimiento mejorando la función en general de la extremidad