Comparing RNA extraction methods to face the variations in RNA quality using two human biological matrices.

BACKGROUND: Nucleic acids, RNA among them, are widely used in biomedicine and Biotechnology. Because of their susceptibility to degradation by RNases, the handling and extraction process of RNA from cells and tissues require specialized personnel and standardized methods to guarantee high purity and integrity. Due to the diversity of techniques found in the market, a comparative study between different RNA extraction methods is useful to facilitate the best choice for the researcher or in research service platforms such as biobanks to see the traceability of the samples. METHODS AND RESULTS: In this study, we have compared seven different RNA extraction methods: manual (TRIzol™), semiautomated (QIAGEN™, Bio-Rad, Monarch®, and Canvax™), and fully automated (QIAcube™ and Maxwell®) processes, from two biological matrices: human Jurkat T cells and peripheral blood mononuclear cells (PBMC). Results showed marked differences in the RNA quality and functionality according to the method employed for RNA extraction and the matrix used. DISCUSSION: QIAcube™ and semi-automated extraction methods were perceived as the best options because of their lower variability, good functionality, and lower cost (P < 0.001). These data contribute to facilitating researchers or research service platforms (Biobanks) in decision-making practices and emphasize the relevance of the selection of the RNA extraction method in each experimental procedure or traceability study to guarantee both quality standards and its reproducibility.

Predictores radiológicos del volumen final del infarto cerebral en pacientes con obstrucción vascular proximal / Radiological predictors of final infarct volume in patients with proximal vascular occlusion

Introducción: Múltiples variables clínicas y radiológicas están involucradas en el pronóstico neurológico de los pacientes con accidente cerebrovascular (ACV) isquémico. Alrededor del 30% de los ACV isquémicos son causados por la obstrucción vascular proximal del circuito anterior; en estos casos, la utilidad de la trombólisis sistémica es limitada. La angiotomografía está indicada en los pacientes que pueden ser candidatos a tratamiento endovascular. Diferentes factores radiológicos como el grado de colaterales leptomeníngeas, o el largo, la densidad o la extensión del trombo, fueron descritos como predictores del pronóstico neurológico tras un ACV isquémico con compromiso vascular proximal. El volumen final del infarto cerebral se correlaciona con la mortalidad y el grado funcional a largo plazo de estos pacientes. El propósito de este estudio es determinar los mejores predictores radiológicos del volumen final del infarto cerebral en pacientes con ACV isquémico con compromiso proximal, utilizando angiotomografía. Materiales y métodos: Realizamos un estudio observacional retrospectivo. Incluimos pacientes adultos con ACV isquémico causado por la obstrucción de un vaso proximal, diagnosticados mediante angiotomografía en el período de junio de 2009 a diciembre de 2019. Medimos la densidad y el largo del trombo en la adquisición sin contraste, registramos el grado de colaterales leptomeníngeas y la extensión del trombo utilizando el clot burden score. Luego medimos el volumen final del infarto en una tomografía de control y analizamos el grado de correlación entre estos factores radiológicos en el volumen infartado. Resultados: Incluimos 54 pacientes con ACV isquémico por compromiso vascular proximal; 41 (75%) fueron mujeres. La mediana de edad fue de 82 años. Alrededor del 60% de los ACV comprometieron el hemisferio derecho y el vaso más afectado fue el segmento M1 de la arteria cerebral media (40,7%)...(AU)

Introduction: Various clinical and radiologic variables impact the neurologic prognosis of patients with ischemic cerebrovascular accidents. About 30% of ischemic cerebrovascular accidents are caused by proximal obstruction of the anterior circulation; in these cases, systemic thrombolysis is of limited usefulness. CT angiography is indicated in candidates for endovascular treatment. Various radiologic factors, including the grade of leptomeningeal collateral circulation, as well as the length, density, and extension of the thrombus, have been identified as predictors of neurologic prognosis after anterior ischemic cerebrovascular accidents due to proximal vascular obstruction. Final infarct volume correlations with mortality and long-term functional outcome in these patients. This study aimed to determine the best predictors of final infarct volume on CT angiography in patients with ischemic cerebral accidents due to proximal occlusion. Materials and methods: This retrospective observational study included adults with ischemic cerebrovascular accidents due to obstruction of the anterior circulation diagnosed by CT angiography in the period comprising June 2009 through December 2019. We measured the length and density of the thrombus in unenhanced CT images, and we used the clot burden score to record the grade of leptomeningeal collateral circulation and the extension of the thrombus. Then we measured the final infarct volume on follow-up CT and analyzed the correlations among these radiologic factors in the infarct volume. Results: We included 54 patients [mean age, 82 y; 41 (75%) women] with ischemic cerebrovascular accidents due to proximal occlusion. About 60% of the cerebrovascular accidents affected the right cerebral hemisphere, and the most commonly affected vessel was the M1 segment of the medial cerebral artery (40.7%)...(AU)

Radiological predictors of final infarct volume in patients with proximal vascular occlusion.

INTRODUCTION: Various clinical and radiologic variables impact the neurologic prognosis of patients with ischemic cerebrovascular accidents. About 30% of ischemic cerebrovascular accidents are caused by proximal obstruction of the anterior circulation; in these cases, systemic thrombolysis is of limited usefulness. CT angiography is indicated in candidates for endovascular treatment. Various radiologic factors, including the grade of leptomeningeal collateral circulation, as well as the length, density, and extension of the thrombus, have been identified as predictors of neurologic prognosis after anterior ischemic cerebrovascular accidents due to proximal vascular obstruction. Final infarct volume correlations with mortality and long-term functional outcome in these patients. This study aimed to determine the best predictors of final infarct volume on CT angiography in patients with ischemic cerebral accidents due to proximal occlusion. MATERIALS AND METHODS: This retrospective observational study included adults with ischemic cerebrovascular accidents due to obstruction of the anterior circulation diagnosed by CT angiography in the period comprising June 2009 through December 2019. We measured the length and density of the thrombus in unenhanced CT images, and we used the clot burden score to record the grade of leptomeningeal collateral circulation and the extension of the thrombus. Then we measured the final infarct volume on follow-up CT and analyzed the correlations among these radiologic factors in the infarct volume. RESULTS: We included 54 patients [mean age, 82 y; 41 (75%) women] with ischemic cerebrovascular accidents due to proximal occlusion. About 60% of the cerebrovascular accidents affected the right cerebral hemisphere, and the most commonly affected vessel was the M1 segment of the medial cerebral artery (40.7%). Final infarct volume correlated with the grade of leptomeningeal collateral circulation (p=0.03) and with the clot burden score (p=0.01). Neither the length nor the density of the thrombus correlated with final infarct volume. CONCLUSION: The final infarct volume can be estimated on the initial CT angiogram. Nevertheless, we found no useful predictive factors in unenhanced CT images. The best independent radiologic predictors of the final infarct volume are the grade of collateral circulation and the clot burden score, especially in patients who did not undergo mechanical thrombectomy, because mechanical thrombectomy improves outcomes. These factors are important for decision making in the management of patients with ischemic cerebrovascular accidents due to proximal occlusion.

Epidemiología de la COVID-19 entre el personal de centros sanitarios y sociosanitarios del área sanitaria de A Coruña y Cee / COVID-19 epidemiology in health and social health workers in the health area of A Coruña and Cee

Objetivo Desde el comienzo de la pandemia ha sido necesario conocer la evolución epidemiológica del SARS-CoV-2. Es por ello que el objetivo de este estudio fue describir las características de la casuística de la COVID-19 en el personal de centros sanitarios y sociosanitarios del área sanitaria de A Coruña y Cee durante la primera ola epidémica, así como determinar la asociación entre el cuadro clínico y/o la duración del mismo y la condición de repositivizar la RT-PCR. Material y métodos En el periodo de estudio se diagnosticaron 210 casos de COVID-19 entre el personal sanitario y sociosanitario del área sanitaria de A Coruña y Cee. Se llevó a cabo un análisis descriptivo de los factores sociodemográficos, así como la búsqueda de asociación entre el cuadro clínico y la duración de la detección de una RT-PCR positiva. Resultados Las categorías profesionales más afectadas fueron enfermería (33,3%) y auxiliares de enfermería (16,2%). El tiempo medio que los casos tardaron en negativizar la RT-PCR fue de 18,3±9,1 días, con una mediana de 17 días. Se observó que 26 casos (13,8%) volvían a obtener un resultado positivo en alguna RT-PCR posterior, sin cumplir criterios de reinfección. La existencia de manifestaciones cutáneas y artralgias se asoció con la repositivización tras ajustar por edad y sexo (OR=4,6 y OR=6,5; respectivamente). Conclusiones En los profesionales sanitarios diagnosticados con COVID-19 durante la primera ola, los síntomas disnea, manifestaciones cutáneas y artralgias determinaron la repositivización de la RT-PCR tras un resultado negativo previo y sin cumplir criterios de reinfección (AU)

Objective Since the beginning of the pandemic, it has become necessary to know the epidemiological evolution of SARS-CoV-2. Therefore, this study aims to describe the characteristics of the casuistry of COVID-19 in health and social-health workers in the health area of A Coruña and Cee during the first epidemic wave, as well as to determine the association between the clinical profile and/or its duration and the condition of RT-PCR repositivization. Materials and methods During the study period, 210 cases belonging to healthcare and social-healthcare workers from the healthcare area of A Coruña and Cee were diagnosed. A descriptive analysis of sociodemographic factors was carried out, as well as the search for association between the clinical picture and the duration of detection of a positive RT-PCR. Results The most affected categories were nursing (33.3%) and nursing assistants (16.2%). The mean time taken for cases to become RT-PCR negative was 18.3±9.1 days, with a median of 17. It was observed that 26 cases (13.8%) had a positive result in a subsequent RT-PCR, without meeting criteria for reinfection. The existence of skin manifestations and arthralgias was associated with repositivization after adjusting for age and sex (OR=4.6 and OR=6.5, respectively). Conclusions In healthcare professionals diagnosed with COVID-19 during the first wave, symptoms such as dyspnea, skin manifestations and arthralgias led to RT-PCR repositivization after a previous negative result and without meeting criteria for reinfection (AU)

RENEB Inter-Laboratory Comparison 2021: The FISH-Based Translocation Assay.

Translocation analysis using fluorescence in situ hybridization (FISH) is the method of choice for dose assessment in case of chronic or past exposures to ionizing radiation. Although it is a widespread technique, unlike dicentrics, the number of FISH-based inter-laboratory comparisons is small. For this reason, although the current Running the European Network of Biological and Physical retrospective Dosimetry (RENEB) inter-laboratory comparison 2021 was designed as a fast response to a real emergency scenario, it was considered a good opportunity to perform an inter-laboratory comparison using the FISH technique to gain further experience. The Bundeswehr Institute of Radiobiology provided peripheral blood samples from one healthy human volunteer. Three test samples were irradiated with blinded doses of 0, 1.2, and 3.5 Gy, respectively. Samples were then sent to the seven participating laboratories. The FISH technique was applied according to the standard procedure of each laboratory. Both, the frequency of translocations and the estimated dose for each sample were sent to the coordinator using a special scoring sheet for FISH. All participants sent their results in due time. However, although it was initially requested to send the results based on the full analysis, evaluating 500 equivalent cells, most laboratories only sent the results based on triage, with a smaller number of analyzed cells. In the triage analysis, there was great heterogeneity in the number of equivalent cells scored. On the contrary, for the full analysis, this number was more homogeneous. For all three samples, one laboratory showed outlier yields compared to the other laboratories. Excluding these results, in the triage analysis, the frequency of translocations in sample no. 1 ranged from 0 to 0.013 translocations per cell, and for samples no. 2 and no. 3 the genomic mean frequency were 0.27 ± 0.03 and 1.47 ± 0.14, with a coefficient of variation of 0.29 and 0.23 respectively. Considering only results obtained in the triage analysis for sample no. 1, all laboratories, except one, classified this sample as the non-irradiated one. For sample no. 2, excluding the outlier value, the mean reported dose was 1.74 ± 0.16 Gy indicating a mean deviation of about 0.5 Gy to the delivered dose of 1.2 Gy. For sample no. 3 the mean dose estimated was 4.21 ± 0.21 Gy indicating a mean deviation of about 0.7 Gy to the delivered dose of 3.5 Gy. In the frame of RENEB, this is the second FISH-based inter-laboratory comparison. The whole exercise was planned as a response to an emergency, therefore, a triage analysis was requested for all the biomarkers except for FISH. Although a full analysis was initially requested for FISH, most of the laboratories reported only a triage-based result. The main reason is that it was not clearly stated what was required before starting the exercise. Results show that most of the laboratories successfully discriminated unexposed and irradiated samples from each other without any overlap. A good agreement in the observed frequencies of translocations was observed but there was a tendency to overestimate the delivered doses. Efforts to improve the harmonization of this technique and subsequent exercises to elucidate the reason for this trend should be promoted.

[COVID-19 epidemiology in health and social health workers in the health area of A Coruña and Cee]. / Epidemiología de la COVID-19 entre el personal de centros sanitarios y sociosanitarios del área sanitaria de A Coruña y Cee.

OBJECTIVE: Since the beginning of the pandemic, it has become necessary to know the epidemiological evolution of SARS-CoV-2. Therefore, this study aims to describe the characteristics of the casuistry of COVID-19 in health and social-health workers in the health area of A Coruña and Cee during the first epidemic wave, as well as to determine the association between the clinical profile and/or its duration and the condition of RT-PCR repositivization. MATERIALS AND METHODS: During the study period, 210 cases belonging to healthcare and social-healthcare workers from the healthcare area of A Coruña and Cee were diagnosed. A descriptive analysis of sociodemographic factors was carried out, as well as the search for association between the clinical picture and the duration of detection of a positive RT-PCR. RESULTS: The most affected categories were nursing (33.3%) and nursing assistants (16.2%). The mean time taken for cases to become RT-PCR negative was 18.3±9.1 days, with a median of 17. It was observed that 26 cases (13.8%) had a positive result in a subsequent RT-PCR, without meeting criteria for reinfection. The existence of skin manifestations and arthralgias was associated with repositivization after adjusting for age and sex (OR=4.6 and OR=6.5, respectively). CONCLUSIONS: In healthcare professionals diagnosed with COVID-19 during the first wave, symptoms such as dyspnea, skin manifestations and arthralgias led to RT-PCR repositivization after a previous negative result and without meeting criteria for reinfection.

Prevalence of Intracranial Aneurysms in Hereditary Hemorrhagic Telangiectasia: Report from a Single Reference Center.

BACKGROUND AND PURPOSE: Neurologic manifestations in hereditary hemorrhagic telangiectasia include an increased incidence of brain abscesses and ischemic strokes due to paradoxic embolization in addition to a wide spectrum of symptoms and complications due to typical brain vascular malformations. Intracranial aneurysms are not part of this brain vascular malformation spectrum. The aim of this study was to determine their prevalence in patients with hereditary hemorrhagic telangiectasia. MATERIALS AND METHODS: This was a single-center, retrospective study. Adult patients from the institutional Hereditary Hemorrhagic Telangiectasia registry with a definitive diagnosis of hereditary hemorrhagic telangiectasia and an available report or angiographic imaging study were included and reviewed to determine the intracranial aneurysm prevalence. In addition, the morphologic characteristics of intracranial aneurysms and possible associated risk factors were collected. RESULTS: Two hundred twenty-eight patients were analyzed. Thirty-seven aneurysms in 33 patients (14.5%; 95% CI, 9.9%-19%) were found. The median diameter of intracranial aneurysms was 3.2 mm (interquartile range, 2.6-4.4 mm). No association between intracranial aneurysm and sex, age, or genetic background was noted. There were no subarachnoid hemorrhagic events due to intracranial aneurysm rupture. CONCLUSIONS: Due to the high prevalence of intracranial aneurysms in adult patients with hereditary hemorrhagic telangiectasia, further studies regarding bleeding risks and monitoring should be addressed.

Reuse of the digestate obtained from the biomethanization of olive mill solid waste (OMSW) as soil amendment or fertilizer for the cultivation of forage grass (Lolium rigidum var. Wimmera).

The principal by-product from the two-phase olive oil production process is olive mill solid waste (OMSW). It is a highly-pollutant by-product, not only because of its characteristics, but also because of the considerable volume of OMSW which is generated, amounting to 2 to 4 million tons per year in Spain. The anaerobic digestion of this by-product is a well-studied process, and results in the generation of biogas, methane and carbon dioxide mainly of high calorific values (20-25 MJ m-3), and an effluent or digestate. The digestate of this by-product has never been characterized. This study presents an informative view on how the composition of OMSW digestate shows promising implications as a soil amendment or fertilizer due to the quality of the biomass from Lolium rigidum, a useful grass specie for the production of forage. Three OMSW digestate alternative applications or treatments were investigated: the digestate and the solid fraction of the digestate for a nutrient-poor soil amendment and the liquid fraction of the digestate as fertilizer. The results confirm that all the OMSW digestate treatments studied presented suitable characteristics for agricultural use, and showed an optimal Carbon/Nitrogen ratio with adequate values for heavy metals which are below the limits established by the Spanish and European legislation in the absence of pathogens. However, fertirrigation was the treatment that provided Lolium rigidum with the best characteristics, improving its shoot biomass, photosynthetic rate and nutritional content.

Biallelic variants in genes previously associated with dominant inheritance: CACNA1A, RET and SLC20A2.

A subset of families with co-dominant or recessive inheritance has been described in several genes previously associated with dominant inheritance. Those recessive families displayed similar, more severe, or even completely different phenotypes to their dominant counterparts. We report the first patients harboring homozygous disease-related variants in three genes that were previously associated with dominant inheritance: a loss-of-function variant in the CACNA1A gene and two missense variants in the RET and SLC20A2 genes, respectively. All patients presented with a more severe clinical phenotype than the corresponding typical dominant form. We suggest that co-dominant or recessive inheritance for these three genes could explain the phenotypic differences from those documented in their cognate dominant phenotypes. Our results reinforce that geneticists should be aware of the possible different forms of inheritance in genes when WES variant interpretation is performed. We also evidence the need to refine phenotypes and inheritance patterns associated with genes in order to avoid failures during WES analysis and thus, raising the WES diagnostic capacity in the benefit of patients.

Validity and reliability of the Child Perceptions Questionnaire 11-14 for Colombian school children.

PURPOSE: To evaluate the validity and reliability of the Child Perceptions Questionnaire 11-14 (CPQ11-14) for children attending public educational institutions in Floridablanca (Colombia). METHODS: An evaluation study of diagnostic technology was conducted. The questionnaire was completed by 500 students aged 11-14 years who were selected by simple random sampling. The test-retest reproducibility was evaluated in 36 students chosen at random from one of the schools; the questionnaire was completed a second time by these students, with an interval of two weeks between administrations. Internal consistency was estimated using the Cronbach's alpha coefficient, and test-retest reproducibility was determined with the intraclass correlation coefficient (ICC). The Rasch Rating Scale Model was used to perform the validation; the structure of the categories, the quality of the items, the invariance and the one-dimensionality were analyzed; invariance was confirmed using differential item functioning (DIF). Informed consent was obtained from the parents or guardians, and assent was obtained from the minors. RESULTS: The mean age was 12.7 ± 1.1 years, and 210 (42.0%) participants were female. The Cronbach's alpha and ICC values were 0.91 and 0.63, respectively. The 37 items fit the Rasch model, but a uniform DIF was observed for an item that was eliminated. The questionnaire was not one-dimensional, and the proportion of variance explained by the model was poor (26.3%). CONCLUSION: The CPQ11-14 showed unsatisfactory test-retest reproducibility and excellent internal consistency; no unidimensionality was observed, but the items fit the Rasch model.

Batch mesophilic anaerobic co-digestion of spent goat batch mesophilic anaerobic co-digestion of spent goat straw bedding and goat cheese whey: Comparison with the mono-digestion of the two sole substrates.

Spent livestock bedding is a valuable resource for the production of green energy (methane) in rural areas. Comparison and evaluation of batch anaerobic digestion and co-digestion of different mixtures of goat straw bedding (SGSB) and goat cheese whey were carried out. Biochemical methane potential (BMP) tests of the 100% SGSB, 95% SGSB-5% whey, 90% SGSB-10% whey, 85% SGSB-15% whey and 100% whey were found to be 423 ± 7, 354 ± 9, 371 ± 2, 293 ± 1, 274 ± 2 mL CH4 g-1 VS. Two different kinetic models were evaluated. The logistic model revealed a decrease in the maximum methane production rate (Rm) from 34.7 ± 1.5 to 14.1 ± 0.9 mL CH4 g-1 VS·d-1 when the percentage of whey in the mixture increased from 0 to 15% as a consequence of the increased ammonia released during the co-digestion of increased concentrations of whey. The lowest value for the maximum methane production predicted by the model (P) was found for 100% whey (274 ± 10 mL CH4 g-1 VS). A two-substrate model was applied to describe the evident existence of rapid and slowly degradable material. Regarding the hydrolysis kinetic constants predicted by this model, considerable increases in the rapid biodegradation stage (krapid) were observed when comparing to the values found for the slow (kslow) biodegradation stage in all the cases tested. The increases between both constants rose from 5 to 42% when the percentage of whey increased.

Influence of Water on the Oxidation of NO on Pd/TiO2 Photocatalysts.

Two series of new photocatalysts were synthesized based on modification with Pd of the commercial P25 photocatalyst (EVONIK®). Two techniques were employed to incorporate Pd nanoparticles on the P25 surface: photodeposition (series Pd-P) and impregnation (series Pd-I). Both series were characterized in depth using a variety of instrumental techniques: BET, DRS, XRD, XPS, TEM, FTIR and FESEM. The modified series exhibited a significant change in pore size distribution, but no differences compared to the original P25 with respect to crystalline phase ratio or particle size were observed. The Pd0 oxidation state was predominant in the Pd-P series, while the presence of the Pd2+ oxidation state was additionally observed in the Pd-I series. The photoactivity tests were performed in a continuous photoreactor with the photocatalysts deposited, by dip-coating, on borosilicate glass plates. A total of 500 ppb of NO was used as input flow at a volumetric flow rate of 1.2 L·min-1, and different relative humidities from 0 to 65% were tested. The results obtained show that under UV-vis or Vis radiation, the presence of Pd nanoparticles favors NO removal independently of the Pd incorporation method employed and independently of the tested relative humidity conditions. This improvement seems to be related to the different interaction of the water with the surface of the photocatalysts in the presence or absence of Pd. It was found in the catalyst without Pd that disproportionation of NO2 is favored through its reaction with water, with faster surface saturation. In contrast, in the catalysts with Pd, disproportionation took place through nitro-chelates and adsorbed NO2 formed from the photocatalytic oxidation of the NO. This different mechanism explains the greater efficiency in NOx removal in the catalysts with Pd. Comparing the two series of catalysts with Pd, Pd-P and Pd-I, greater activity of the Pd-P series was observed under both UV-vis and Vis radiation. It was shown that the Pd0 oxidation state is responsible for this greater activity as the Pd-I series improves its activity in successive cycles due to a reduction in Pd2+ species during the photoactivity tests.

Deficits in coordinated neuronal activity and network topology are striatal hallmarks in Huntington's disease.

BACKGROUND: Network alterations underlying neurodegenerative diseases often precede symptoms and functional deficits. Thus, their early identification is central for improved prognosis. In Huntington's disease (HD), the cortico-striatal networks, involved in motor function processing, are the most compromised neural substrate. However, whether the network alterations are intrinsic of the striatum or the cortex is not fully understood. RESULTS: In order to identify early HD neural deficits, we characterized neuronal ensemble calcium activity and network topology of HD striatal and cortical cultures. We used large-scale calcium imaging combined with activity-based network inference analysis. We extracted collective activity events and inferred the topology of the neuronal network in cortical and striatal primary cultures from wild-type and R6/1 mouse model of HD. Striatal, but not cortical, HD networks displayed lower activity and a lessened ability to integrate information. GABAA receptor blockade in healthy and HD striatal cultures generated similar coordinated ensemble activity and network topology, highlighting that the excitatory component of striatal system is spared in HD. Conversely, NMDA receptor activation increased individual neuronal activity while coordinated activity became highly variable and undefined. Interestingly, by boosting NMDA activity, we rectified striatal HD network alterations. CONCLUSIONS: Overall, our integrative approach highlights striatal defective network integration capacity as a major contributor of basal ganglia dysfunction in HD and suggests that increased excitatory drive may serve as a potential intervention. In addition, our work provides a valuable tool to evaluate in vitro network recovery after treatment intervention in basal ganglia disorders.

Morphologic Features on MR Imaging Classify Multifocal Glioblastomas in Different Prognostic Groups.

BACKGROUND AND PURPOSE: Multifocal glioblastomas (ie, glioblastomas with multiple foci, unconnected in postcontrast pretreatment T1-weighted images) represent a challenge in clinical practice due to their poor prognosis. We wished to obtain imaging biomarkers with prognostic value that have not been found previously. MATERIALS AND METHODS: A retrospective review of 1155 patients with glioblastomas from 10 local institutions during 2006-2017 provided 97 patients satisfying the inclusion criteria of the study and classified as having multifocal glioblastomas. Tumors were segmented and morphologic features were computed using different methodologies: 1) measured on the largest focus, 2) aggregating the different foci as a whole, and 3) recording the extreme value obtained for each focus. Kaplan-Meier, Cox proportional hazards, correlations, and Harrell concordance indices (c-indices) were used for the statistical analysis. RESULTS: Age (P < .001, hazard ratio = 2.11, c-index = 0.705), surgery (P < .001, hazard ratio = 2.04, c-index = 0.712), contrast-enhancing rim width (P < .001, hazard ratio = 2.15, c-index = 0.704), and surface regularity (P = .021, hazard ratio = 1.66, c-index = 0.639) measured on the largest focus were significant independent predictors of survival. Maximum contrast-enhancing rim width (P = .002, hazard ratio = 2.05, c-index = 0.668) and minimal surface regularity (P = .036, hazard ratio = 1.64, c-index = 0.600) were also significant. A multivariate model using age, surgery, and contrast-enhancing rim width measured on the largest foci classified multifocal glioblastomas into groups with different outcomes (P < .001, hazard ratio = 3.00, c-index = 0.853, median survival difference = 10.55 months). Moreover, quartiles with the highest and lowest individual prognostic scores based on the focus with the largest volume and surgery were identified as extreme groups in terms of survival (P < .001, hazard ratio = 18.67, c-index = 0.967). CONCLUSIONS: A prognostic model incorporating imaging findings on pretreatment postcontrast T1-weighted MRI classified patients with glioblastoma into different prognostic groups.

Rescue of Discarded Grafts for Liver Transplantation by Ex Vivo Subnormothermic and Normothermic Oxygenated Machine Perfusion: First Experience in Spain.

BACKGROUND: Ex vivo machine perfusion (MP) has been reported as a possibly method to rescue discarded organs. The main aim of this study was to report an initial experience in Spain using MP for the rescue of severely marginal discarded liver grafts, and to, secondarily, define markers of viability to test the potential applicability of these devices for the real increase in the organ donor pool. METHODS: The study began in January 2016. Discarded grafts were included in a research protocol that consisted of standard retrieval followed by 10 hours of cold ischemia. Next, either normothermic (NMP) or controlled subnormothermic (subNMP) rewarming was chosen randomly. Continuous measurements of portal-arterial pressure and resistance were screened. Lactate, pH, and bicarbonate were measured every 30 minutes. The perfusion period was 6 hours, after which the graft was discarded and evaluated as potentially usable, but never implanted. Biopsies of the donor and at 2, 4, and 6 hours after ex vivo MP were obtained. RESULTS: A total of 4 grafts were included in the protocol. The first 2 grafts were perfused by NMP and grafts 3 and 4 by subNMP. The second and third grafts showed a clear trend toward optimal recovery and may have been used. Lactate dropped to levels below 2.5 mmol/L with stable arterial and portal pressure and resistance. Clear biliary output started during MP. Biopsies showed an improvement of liver architecture with reduced inflammation at the end of the perfusion. CONCLUSION: This preliminary experience has demonstrated the potential of MP devices for the rescue of severely marginal liver grafts. Lactate and biliary output were useful for viability testing of the grafts. The utility of NMP or subNMP protocols requires further research.

Participative approach to elicit water quality monitoring needs from stakeholder groups - An application of integrated watershed management.

Water quality monitoring programs (WQMPs) must be based on monitoring objectives originating from the real knowledge needs of all stakeholders in a watershed and users of the resource. This paper proposes a participative approach to elicit knowledge needs and preferred modes of communication from citizens and representatives of organized stakeholders (ROS) on water quality and quantity issues. The participative approach includes six steps and is adaptable and transferable to different types of watersheds. These steps are: (1) perform a stakeholder analysis; (2) conduct an adaptable survey accompanied by a user-friendly public participation geographical information system (PPGIS); (3) hold workshops to meet with ROS to inform them of the results of the survey and PPGIS; discuss attainment of past monitoring objectives; exchange views on new knowledge needs and concerns on water quality and quantity; (4) meet with citizens to obtain the same type of input (as from ROS); (5) analyze the data and information collected to identify new knowledge needs and modes of communication and (6) identify, in collaboration with the individuals in charge of the WQMPs, the short-, medium- and long-term monitoring objectives and communication strategies to be pursued. The participative approach was tested on two distinct watersheds in the province of Quebec, Canada. It resulted in a series of optimization objectives of the existing WQMPs, new monitoring objectives and recommendations regarding communication strategies of the WQMPs' results. The results of this study show that the proposed methodology is appreciated by all parties and that the outcomes and monitoring objectives are acceptable. We also conclude that successful integrated watershed management is a question of scale, and that every aspect of integrated watershed management needs to be adapted to the surface watershed, the groundwater watershed (aquifers) and the human catchment area.

Dermatophagoides pteronyssinus immunotherapy changes the T-regulatory cell activity.

Subcutaneous specific immunotherapy (SCIT) has been shown to modify the Dermatophagoides pteronissinus (DP) allergic response, characterized by generation of Treg cells. However, studies have reported no changes in the proportion of Treg cells after immunotherapy, indicating that the effects may be due to modifications in their regulatory activities. We aimed to determine whether Tregs generated by DP-SCIT can switch the allergic response to tolerant and study the involvement of suppressive cytokines on it. Twenty-four DP-allergic rhinitis patients were recruited, 16 treated with DP-SCIT and 8 untreated. Treg and T effector cells were isolated before and after DP-SCIT, and cocultured in different combinations with α-IL-10, α-TGF-ß blocking antibodies and nDer p 1. Treg cells after DP-SCIT increased Th1 and decreased Th2 and Th9 proliferation. Similarly, they increased IL-10 and decreased IL-4 and IL-9-producing cells. α-IL-10 affected the activity of Treg cells obtained after DP-SCIT only. Finally, DP-specific IgG4 levels, Treg percentage and IL-10 production were correlated after DP-SCIT. These results demonstrate that DP-SCIT induces Treg cells with different suppressive activities. These changes could be mediated by IL-10 production and appear to play an important role in the induction of the tolerance response leading to a clinical improvement of symptoms.

Biodegradable Porous Silk Microtubes for Tissue Vascularization.

Cardiovascular diseases are the leading cause of mortality around the globe, and microvasculature replacements to help stem these diseases are not available. Additionally, some vascular surgeries needing small diameter vascular grafts present different performance requirements. In this work silk fibroin scaffolds based on silk/polyethylene oxide blends were developed as microtubes for vasculature needs and for different tissue regeneration times, mechanical properties and structural designs. Systems with 13, 14 and 15% silk alone or blended with 1 or 2% of polyethylene oxide (PEO) were used to generate porous microtubes using gel-spinning. Microtubes with inner diameters (ID) of 150-300 µm and 100 µm wall thickness were fabricated. The systems were assessed for porosity, mechanical properties, enzymatic degradability, and in vitro vascular endothelial cell attachment and metabolic activity. After 14 days all tubes supported the proliferation of cells and cell attachment increased with porosity. The silk tubes with PEO had similar crystallinity but higher elastic modulus compared with the systems without PEO. The silk (13%)/PEO (1%) system showed the highest porosity (20 µm pore diameters on average), highest cell attachment and fastest degradation profile. There was a good correlation between these parameters with silk concentration and the presence of PEO. The results demonstrate the ability to generate versatile and tunable tubular biomaterials based on silk-PEO-blends with potential for microvascular grafts.

Repair of nerve injury by implanting prostheses obtained from isogenic acellular nerve segments. / Reparación de lesiones en nervios mediante el implante de prótesis obtenidas de segmentos acelulares de nervio isogénico.

INTRODUCTION: When a nerve section with a significant gap occurs, it is necessary to use a prosthesis to suture it. To date an autologous nerve segment graft appears to be the best treatment; but it has several important disadvantages. Our goal is to study the effectiveness of an isogenic acellular nerve prosthesis comparing a simple suture with tubulisation. MATERIAL AND METHOD: Four groups of Wistar rats were used. The animals in Group 0 served as donors of nerve segments to graft. Group 1 received the implant with an end-to-end suture. In group 2, the implant was sutured inside an ɛ-caprolactone tube. Group 3 received it in a polylactic-co-glycolic acid tube. We evaluated the motor function (sciatic index and step test in motion), and the regeneration length by histological study of regeneration, after a maximum of 3 weeks. RESULTS: Regeneration was uneven in the three groups. In all groups, there were implants with regenerated nerve fibres at the maximum studied length (15mm) and others where regeneration was scarce. The mean regeneration length was greater in the direct end-to-end suture group (G1), although the regeneration speed was similar in the three groups. Group 1 showed the highest percentage of regeneration, but the variability of results prevents this difference reaching statistical significance. We found no significant differences between the two groups with polymer tubes. CONCLUSION: For the implantation of isogenic acellular nerve prosthesis, under our experimental conditions, the direct end-to-end suture was more effective than when it isprotected with biopolymer tubes.

Prevalence and distribution of hepatitis C virus genotypes in Spain during the 2000-2015 period (the GEHEP 005 study).

We report the largest study on the prevalence and distribution of HCV genotypes in Spain (2000-2015), and we relate them with clinical, epidemiological and virological factors. Patients from 29 hospitals in 10 autonomous communities (Andalusia, Aragon, Castilla-Leon, Catalonia, Galicia, Canary Islands, Madrid Community, Valencian Community, Murcia Region and Basque Country) have been studied. Annual distribution of HCV genotypes and subtypes, as well as gender, age, transmission route, HIV and/or HBV coinfection, and treatment details were recorded. We included 48595 chronically HCV-infected patients with the following characteristics: median age 51 years (IQR, 44-58), 67.9% male, 19.1% HIV-coinfected, 23.5% HBV-coinfected. Parenteral transmission route was the most frequent (58.7%). Genotype distribution was 66.9% GT1 (24.9% subtype 1a and 37.9% subtype 1b), 2.8% GT2, 17.3% GT3, 11.4% GT4 and 0.1% GT5 and 0.02% GT6. LiPA was the most widely HCV genotyping test used (52.4%). HCV subtype 1a and genotypes 3 and 4 were closely associated with male gender, parenteral route of infection and HIV and HBV coinfection; in contrast, subtype 1b and genotype 2 were associated with female gender, nonparenteral route and mono-infection. Age was related to genotype distribution, and different patterns of distribution and biodiversity index were observed between different geographical areas. Finally, we describe how treatment and changes in transmission routes may have affected HCV genotype prevalence and distribution patterns. We present the most recent data on molecular epidemiology of hepatitis C virus in Spain. This study confirms that genotype distributions vary with age, sex, HIV and HBV coinfection and within geographical areas and epidemiological groups.