Effects of Blood flow Restriction and Load on Mean Propulsive Velocity and Subjective Perceived Exertion During Squat and Bench Press Exercises.

BACKGROUND: The aim of this study was to determine the influence of different percentages of blood flow restriction (BFR) and loads on mean propulsive velocity (MPV) and subjective perceived exertion during squat (SQ) and bench press (BP) exercises. HYPOTHESIS: Higher percentages of BFR will positively affect dependent variables, increasing MPV and reducing perceived exertion. STUDY DESIGN: Cross-sectional study. LEVEL OF EVIDENCE: Level 3. METHODS: Eight healthy young male athletes took part. Two sets of 6 repetitions at 70% 1-repetition maximum (1RM), 2 sets of 4 repetitions at 80% 1RM, and 2 sets of 2 repetitions at 90% 1RM were performed randomly; 5-minute recoveries were applied in all sets. The varying arterial occlusion pressure (AOP) applied randomly was 0% (Control [CON]), 80%, and 100%. RESULTS: No statistically significant differences in MPV were found during the BP exercise at any percentage of BFR at any percentage 1RM. During the SQ exercise, MPV results showed statistically significant increases of 5.46% (P = 0.04; ηp2 = 0.31) between CON and 100% AOP at 90% 1RM. The perceived exertion results for the BP exercise showed statistically significant reductions of -8.66% (P < 0.01; ηp2 = 0.06) between CON and 100% AOP at 90% 1RM. During the SQ exercise, the perceived exertion results showed significant reductions of -10.04% (P = 0.04; ηp2 = 0.40) between CON and 100% AOP at 80% 1RM; -5.47% (P = 0.02; ηp2 = 0.48) between CON and 80% AOP at 90% 1RM; and -11.83% (P < 0.01; ηp2 = 0.66) between CON and 100% AOP at 90% 1RM. CONCLUSION: BFR percentages ~100% AOP at 90% 1RM improved acutely MPV (only in SQ exercises) and reduced acutely perceived exertion (in both exercises). These findings are important to consider when prescribing resistance training for healthy male athletes.

Endosperm Persistence in Arabidopsis Results in Seed Coat Fractures and Loss of Seed Longevity.

Seeds are specialized plant organs that carry, nurture, and protect plant offspring. Developmental coordination between the three genetically distinct seed tissues (the embryo, endosperm, and seed coat) is crucial for seed viability. In this study, we explore the relationship between the TFs AtHB25 and ICE1. Previous results identified ICE1 as a target gene of AtHB25. In seeds, a lack of ICE1 (ice1-2) suppresses the enhanced seed longevity and impermeability of the overexpressing mutant athb25-1D, but surprisingly, seed coat lipid polyester deposition is not affected, as shown by the double-mutant athb25-1D ice1-2 seeds. zou-4, another mutant lacking the transcriptional program for proper endosperm maturation and for which the endosperm persists, also presents a high sensitivity to seed aging. Analysis of gso1, gso2, and tws1-4 mutants revealed that a loss of embryo cuticle integrity does not underlie the seed-aging sensitivity of ice1-2 and zou-4. However, scanning electron microscopy revealed the presence of multiple fractures in the seed coats of the ice1 and zou mutants. Thus, this study highlights the importance of both seed coat composition and integrity in ensuring longevity and demonstrates that these parameters depend on multiple factors.

Kaempferol-3-rhamnoside overaccumulation in flavonoid 3'-hydroxylase tt7 mutants compromises seed coat outer integument differentiation and seed longevity.

Seeds slowly accumulate damage during storage, which ultimately results in germination failure. The seed coat protects the embryo from the external environment, and its composition is critical for seed longevity. Flavonols accumulate in the outer integument. The link between flavonol composition and outer integument development has not been explored. Genetic, molecular and ultrastructural assays on loss-of-function mutants of the flavonoid biosynthesis pathway were used to study the effect of altered flavonoid composition on seed coat development and seed longevity. Controlled deterioration assays indicate that loss of function of the flavonoid 3' hydroxylase gene TT7 dramatically affects seed longevity and seed coat development. Outer integument differentiation is compromised from 9 d after pollination in tt7 developing seeds, resulting in a defective suberin layer and incomplete degradation of seed coat starch. These distinctive phenotypes are not shared by other mutants showing abnormal flavonoid composition. Genetic analysis indicates that overaccumulation of kaempferol-3-rhamnoside is mainly responsible for the observed phenotypes. Expression profiling suggests that multiple cellular processes are altered in the tt7 mutant. Overaccumulation of kaempferol-3-rhamnoside in the seed coat compromises normal seed coat development. This observation positions TRANSPARENT TESTA 7 and the UGT78D1 glycosyltransferase, catalysing flavonol 3-O-rhamnosylation, as essential players in the modulation of seed longevity.

Acute effects of different external compression with blood flow restriction on force-velocity profile during squat and bench press exercises.

The aim was to compare the acute effects of bench press (BP) and squat (SQ) exercises with blood flow restriction (BFR) (40%, 60%, 80% and 100% of the complete arterial occlusion pressure (AOP)) and without BFR (CON) on the mean propulsive (VelMED) and maximum (VelMAX) bar velocity. Fourteen healthy, physically active males (age, 23.6 ± 4.1 years; height, 1.85 ± 0.11 m; body weight 85.4 ± 4.1 kg) took part in the study. There was one set for each testing condition (CON, 40%, 60%, 80% and 100%) with 6 repetitions for BP and 6 repetitions for SQ, at 60% of 1RM, and 3 minutes of recovery between sets. The results showed statistically significant differences of the sets with 80% BFR vs. CON (mean difference [MD] = 0.035 m · s-1, p < 0.05, ES = 0.52 [1.02-0.03]) and 100% BFR sets vs. CON (MD = 0.074, p < 0.001, ES = 1.08 [1.79-0.38]) for BP. In the SQ exercise, statistically significant differences were found between 100% BFR vs. CON (DM = 0.031 m · s-1, p < 0.05), vs. 100% BFR 40% (MD = 0.04 m · s-1, p < 0.05). Trend analysis showed a statistically significant linear trend (F[1,9] = 34.9, p < 0.001, F[1,13] = 27.32, p < 0.001) for the VelMED in relation to the different levels of BFR. In conclusion, our results showed that BFR levels above ˜80% AOP (BP) and ˜100% AOP (SQ) produce a VelMED improvement at 60% 1RM.

Quistes óseos aneurismáticos. Terapia celular con células mononucleares y seguimiento imagenológico en dos pacientes adolescentes / Aneurysmal bone cysts. Mononuclear cell therapy and imaging follow-up in two adolescent patients

RESUMEN Los quistes óseos aneurismáticos son frecuentes en la edad pediátrica. Para su determinación se cuenta con diversos estudios imagenológicos como la radiografía y la tomografía axial computarizada que pueden colaborar al diagnóstico diferencial con otras lesiones. Existen disímiles opciones terapéuticas, el uso de factores de crecimiento autólogos se ha considerado como alternativa eficaz. Se presentan dos pacientes consultados en el servicio de Ortopedia del Hospital Provincial Pediátrico Universitario «José Luis Miranda¼, de Villa Clara, con diagnóstico clínico e imagenológico de quiste óseo aneurismático que recibieron tratamiento mediante terapia celular con células mononucleares con buena evolución clínica y radiográfica. Esta técnica es aplicable en nuestro medio ya que no requiere de estimables recursos materiales lo que constituye una fortaleza para su implementación. Las radiografías permiten reconocer la evolución posterior al tratamiento.

ABSTRACT Aneurysmal bone cysts are common in children. For its determination, various imaging studies are available, such as radiography and computerized axial tomography, which can collaborate in the differential diagnosis with other lesions. There are dissimilar therapeutic options, the use of autologous growth factors has been considered as an effective alternative. We present two patients seen in the Orthopedics service at "José Luis Miranda" University Pediatric Hospital in Villa Clara, with a clinical and imaging diagnosis of aneurysmal bone cyst who received treatment with mononuclear cell therapy having a good clinical and radiographic evolution. This technique is applicable in our environment since it does not require considerable material resources, which constitutes a strength for its implementation. X-rays allow us to recognize the evolution after treatment.

Transcription Factor DOF4.1 Regulates Seed Longevity in Arabidopsis via Seed Permeability and Modulation of Seed Storage Protein Accumulation.

Seed longevity is modulated by multiple genetic factors in Arabidopsis thaliana. A previous genome-wide association study using the Elevated Partial Pressure of Oxygen (EPPO) aging assay pinpointed a genetic locus associated with this trait. Reverse genetics identified the transcription factor DOF4.1 as a novel seed longevity factor. dof4.1 loss-of-function plants generate seeds exhibiting higher germination after accelerated aging assays. DOF4.1 is expressed during seed development and RNAseq data show several putative factors that could contribute to the dof4.1 seed longevity phenotype. dof4.1 has reduced seed permeability and a higher levels of seed storage proteins mRNAs (cruciferins and napins) in developing seeds, as compared to wild-type seeds. It has been reported that mutant lines defective in cruciferins or napins present reduced seed longevity. The improved longevity of dof4.1 is totally lost in the quadruple mutant dof4.1 cra crb crc, but not in a dof4.1 line depleted of napins, suggesting a prominent role for cruciferins in this process. Moreover, a negative regulation of DOF4.1 expression by the transcription factor DOF1.8 is suggested by co-inoculation assays in Nicotiana benthamiana. Indeed, DOF1.8 expression anticorrelates with that of DOF4.1 during seed development. In summary, modulation of DOF4.1 levels during seed development contributes to regulate seed longevity.

Comparative analysis of wild-type accessions reveals novel determinants of Arabidopsis seed longevity.

Understanding the genetic factors involved in seed longevity is of paramount importance in agricultural and ecological contexts. The polygenic nature of this trait suggests that many of them remain undiscovered. Here, we exploited the contrasting seed longevity found amongst Arabidopsis thaliana accessions to further understand this phenomenon. Concentrations of glutathione were higher in longer-lived than shorter-lived accessions, supporting that redox poise plays a prominent role in seed longevity. However, high seed permeability, normally associated with shorter longevity, is also present in long-lived accessions. Dry seed transcriptome analysis indicated that the contribution to longevity of stored messenger RNA (mRNAs) is complex, including mainly accession-specific mechanisms. The detrimental effect on longevity caused by other factors may be counterbalanced by higher levels of specific mRNAs stored in dry seeds, for instance those of heat-shock proteins. Indeed, loss-of-function mutant analysis demonstrated that heat-shock factors HSF1A and 1B contributed to longevity. Furthermore, mutants of the stress-granule zinc-finger protein TZF9 or the spliceosome subunits MOS4 or MAC3A/MAC3B, extended seed longevity, positioning RNA as a novel player in the regulation of seed viability. mRNAs of proteins with putative relevance to longevity were also abundant in shorter-lived accessions, reinforcing the idea that resistance to ageing is determined by multiple factors.

Seed coat lignification level is crucial in Capsicum spp seed longevity.

Capsicum (pepper) is known for its poor seed germination, particularly seed longevity is usually much shorter than other Solanaceae. However, the molecular mechanisms involved are mostly unknown in these species. The present study examines the differences in seed longevity among Capsicum species and varietal types. Feral or less domesticated species, such as Capsicum chinense and particularly Capsicum frutescens, showed higher germination rates than the more domesticated Capsicum annuum after accelerated seed aging treatments. In addition, variability was detected in the expression of genes involved in the response to seed deterioration. The differences observed in ASPG1 expression led us to study the seed protein profile in dry and germinating seeds. Seed storage protein mobilization during germination was faster in seed aging-resistant genotypes. Similarly, the transcriptional change observed for the orthologous gene of the trans-species regulator AtHB25 prompted us to study the structure and molecular components of the seed coat in peppers. All the Capsicum pepper accessions analyzed presented very lignified testa and we observed a positive correlation between the amount of lignin and seed viability. Our results provide essential information to explain the poor germination observed in pepper seeds and provide an experimental framework for future improvements in this important character.

Restauración de la biomecánica en artroplastia total de cadera por luxación congénita alta / Restoration of Biomechanics in Total Hip Arthroplasty Due to High Congenital Dislocation

La osteoartritis secundaria a displasia o luxación congénita de cadera conlleva grandes dificultades para el cirujano que reconstruye la cadera. Los casos con escasa deformidad no difieren prácticamente de la reconstrucción primaria convencional. En el extremo opuesto están los casos con graves hipoplasias del acetábulo, escaso desarrollo femoral, luxación completa, discrepancia importante de las extremidades y gran cabalgamiento del trocánter mayor. Por las enormes dificultades que pueden representar para la cirugía, nos trazamos el objetivo de discutir nuestro caso, con las consideraciones y resultados del tratamiento elegido. Se presenta paciente femenina de 54 años de edad, con antecedentes de salud previa, que nos llegó a consulta con una grave deformidad congénita, acortamiento del miembro inferior derecho (6 cm) y limitación dolorosa de todos los movimientos de la cadera. Se constata una luxación congénita grado C de Hartofilakidis y IV de Crowe, que muestra como parte del tratamiento, la artroplastia total con injerto autólogo y reimplantación del cótilo en el acetábulo verdadero, para recuperar el centro de rotación del acetábulo y la osteotomía femoral de acortamiento para la implantación del vástago femoral. Se exponen los requerimientos, procederes técnicos y resultados alcanzados(AU)

Osteoarthritis secondary to congenital hip dysplasia or dislocation poses great difficulties for the surgeon reconstructing the hip. Cases with little deformity do not differ practically from conventional primary reconstruction. At the opposite end there are cases with severe acetabulum hypoplasia, poor femoral development, complete dislocation, significant limb discrepancy, and great thrust of the greater trochanter. Due to the enormous difficulties that they can represent for surgery, we set the objective of discussing our case, with the considerations and results of the chosen treatment. A 54-year-old female patient is reported, she has previous health history, and she came for consultation with severe congenital deformity, shortening of her right lower limb (6 cm) and painful limitation of all hip movements. Congenital dislocation grade C of Hartofilakidis and IV of Crowe was confirmed, which showed as part of the treatment, the total arthroplasty with autologous graft and reimplantation of the cup in the true acetabulum, to recover the center of rotation of the acetabulum and the shortening femoral osteotomy for implantation of the femoral stem. Requirements, technical procedures and results achieved are informed(AU)

Componentes protésicos más usados en la artroplastia total de cadera en las provincias centrales / Most Used Prosthetic Components in Total Hip Arthroplasty in Central Provinces

Introducción: La cirugía de la artroplastia y el recambio de una prótesis de cadera es una cirugía compleja y costosa. La estrategia quirúrgica y componentes protésicos son completamente diferentes en función de las deformidades óseas existentes y características anatómicas personales de cada paciente, por tanto, la planificación de suministros de los diferentes componentes es básica para cubrir las demandas de la población necesitada y la adecuada eficiencia económica para evitar gastos innecesarios por sobreestimación o infraestimación de las demandas. Objetivos: Determinar las necesidades por medidas de los diferentes tipos de componentes protésicos utilizados en la artroplastia total de cadera en las provincias centrales y de este modo optimizar las inversiones, con las ventajas económicas que supone. Métodos: Estudio longitudinal descriptivo retrospectivo de los casos operados de artroplastia total de cadera en el Hospital Arnaldo Milián Castro de Villa Clara, desde diciembre de 1998 hasta la fecha. Se analizan las variables edad, sexo, procedencia y componentes protésicos. Se recoge la información en la base de datos creada en SPSS, donde se realizan los análisis estadísticos. Resultados: La edad media fue 62,36 años. El grupo de 61 - 80 años fue el de mayor incidencia con 215 pacientes. Predominó el sexo masculino 51,8 por ciento. La mayor incidencia, correspondió a Villa Clara con 692 pacientes. Los componentes cementados 46 - 50 fueron los más usados y en los no cementados los más usados fueron 48-56. Vástagos Tipo Müller 7.5 - 12.5. RALCA Cementadas T0-T4, no cementados T1-T3. En las SLA, 11.5 y 13. Los cuellos estándar fueron los de mayor incidencia con 390 pacientes. Conclusiones: Con las incidencias de las diferentes medidas utilizadas por cada uno de los componentes expresados, se puede determinar con bastante exactitud la demanda de cada medida por cada 100 casos pendientes de operación. Esto permite satisfacer las demandas de las medidas más usadas y hacer una distribución apropiada a la hora del suministro, lo que redundaría en un beneficio económico vital en las actuales condiciones del país(AU)

Introduction: The arthroplasty surgery and the replacement of a hip replacement is a complex and expensive surgery. The surgical strategy and prosthetic components are completely different depending on the existing bone deformities and personal anatomical characteristics of each patient, therefore, the planning of supplies of the different components is essential to meet the demands of the needy population and adequate economic efficiency to avoid unnecessary expenses due to overestimation or underestimation of claims. Objective: To determine the needs by measurements of the different types of prosthetic components used in total hip arthroplasty in the central provinces and thus optimize investments, with the economic advantages that it entails. Methods: A retrospective descriptive longitudinal study was carried out of the total hip arthroplasty operated cases at Arnaldo Milián Castro Hospital in Villa Clara, from December 1998 to date. Age, sex, origin and prosthetic components are the variables examined. The information is collected in the database created in SPSS, statistical analyzes was performed. Results: The mean age was 62.36 years. The group of 61-80 years was the one with the highest incidence (215 patients). Male sex predominated (51.8 percent). The highest incidence corresponded to Villa Clara (692 patients). The cemented components 46-50 were the most used and in the uncemented components the most used were 48-56. Müller Stems 7.5 - 12.5. RALCA Cemented T0-T4, uncemented T1-T3. In the SLA the most used were 11.5 and 13 respectively. Standard necks were those with the highest incidence (390 patients). Conclusions: The demand for each measure can be determined quite accurately for every 100 cases pending operation with the incidents of the different measures used by each of the components expressed, allowing to meet the demands of the most used measures and appropriate distribution at supplying, which would result in vital economic benefit in the current conditions of the country(AU)

Impact of intermediate to high doses of methylprednisolone on mortality rate in patients with COVID-19 pneumonia-induced severe systemic inflammation.

INTRODUCTION: In addition to respiratory support needs, patients' characteristics to guide indication or timing of corticosteroid treatment in COVID-19 patients are not completely established. This study aimed to evaluate the impact of methylprednisolone on mortality rate in patients with COVID-19 pneumonia-induced severe systemic inflammation (PI-SSI). METHODS: Between 9 March and 5 May 2020 (final follow-up on 2 July 2020), a retrospective cohort study was conducted in hospitalised patients with COVID-19 PI-SSI (≥2 inflammatory biomarkers [IBs]: temperature ≥38℃, lymphocyte ≤800 cell/µL, C-reactive protein ≥100 mg/L, lactate dehydrogenase ≥300 units/L, ferritin ≥1000 mcg/L, D-dimer ≥500 ng/mL). Patients received 0.5-1.0 mg/kg of methylprednisolone for 5-10 days or standard of care. The primary outcome was 28-day all-cause mortality. Secondary outcomes included ≥2 points improvement on a 7-item WHO-scale (Day 14), transfer to intensive care unit (ICU) (Day 28) and adverse effects. Kaplan-Meier method and Cox proportional hazard regression were implemented to analyse the time to event outcomes. RESULTS: A total of 142 patients (corticosteroid group n = 72, control group n = 70) were included. A significant reduction in 28-day all-cause mortality was shown with methylprednisolone in patients with respiratory support (HR: 0.15; 95% CI 0.03-0.71), with ≥3 (HR: 0.17; 95% CI 0.05-0.61) or ≥4 altered IB (HR: 0.15; 95% CI 0.04-0.54) and in patients with both respiratory support and ≥3 (HR: 0.11; 95% CI 0.02-0.53] or ≥4 altered IB (HR: 0.14; 95% CI 0.04-0.51). No significant differences were found in secondary outcomes. CONCLUSION: Intermediate to high doses of methylprednisolone, initiated between 5 and 12 days after symptom onset, was associated with a significant reduction in 28-day all-cause mortality in patients with COVID-19 pneumonia and ≥3 o ≥ 4 altered IB, independently of the need of respiratory support.

Apoplastic lipid barriers regulated by conserved homeobox transcription factors extend seed longevity in multiple plant species.

Cutin and suberin are lipid polyesters deposited in specific apoplastic compartments. Their fundamental roles in plant biology include controlling the movement of gases, water and solutes, and conferring pathogen resistance. Both cutin and suberin have been shown to be present in the Arabidopsis seed coat where they regulate seed dormancy and longevity. In this study, we use accelerated and natural ageing seed assays, glutathione redox potential measures, optical and transmission electron microscopy and gas chromatography-mass spectrometry to demonstrate that increasing the accumulation of lipid polyesters in the seed coat is the mechanism by which the AtHB25 transcription factor regulates seed permeability and longevity. Chromatin immunoprecipitation during seed maturation revealed that the lipid polyester biosynthetic gene long-chain acyl-CoA synthetase 2 (LACS2) is a direct AtHB25 binding target. Gene transfer of this transcription factor to wheat and tomato demonstrated the importance of apoplastic lipid polyesters for the maintenance of seed viability. Our work establishes AtHB25 as a trans-species regulator of seed longevity and has identified the deposition of apoplastic lipid barriers as a key parameter to improve seed longevity in multiple plant species.

Uncovering salt tolerance mechanisms in pepper plants: a physiological and transcriptomic approach.

BACKGROUND: Pepper is one of the most cultivated crops worldwide, but is sensitive to salinity. This sensitivity is dependent on varieties and our knowledge about how they can face such stress is limited, mainly according to a molecular point of view. This is the main reason why we decided to develop this transcriptomic analysis. Tolerant and sensitive accessions, respectively called A25 and A6, were grown for 14 days under control conditions and irrigated with 70 mM of NaCl. Biomass, different physiological parameters and differentially expressed genes were analysed to give response to differential salinity mechanisms between both accessions. RESULTS: The genetic changes found between the accessions under both control and stress conditions could explain the physiological behaviour in A25 by the decrease of osmotic potential that could be due mainly to an increase in potassium and proline accumulation, improved growth (e.g. expansins), more efficient starch accumulation (e.g. BAM1), ion homeostasis (e.g. CBL9, HAI3, BASS1), photosynthetic protection (e.g. FIB1A, TIL, JAR1) and antioxidant activity (e.g. PSDS3, SnRK2.10). In addition, misregulation of ABA signalling (e.g. HAB1, ERD4, HAI3) and other stress signalling genes (e.g. JAR1) would appear crucial to explain the different sensitivity to NaCl in both accessions. CONCLUSIONS: After analysing the physiological behaviour and transcriptomic results, we have concluded that A25 accession utilizes different strategies to cope better salt stress, being ABA-signalling a pivotal point of regulation. However, other strategies, such as the decrease in osmotic potential to preserve water status in leaves seem to be important to explain the defence response to salinity in pepper A25 plants.

Identification of novel seed longevity genes related to oxidative stress and seed coat by genome-wide association studies and reverse genetics.

Seed longevity is a polygenic trait of relevance for agriculture and for understanding the effect of environment on the ageing of biological systems. In order to identify novel longevity genes, we have phenotyped the natural variation of 270 ecotypes of the model plant, Arabidopsis thaliana, for natural ageing and for three accelerated ageing methods. Genome-wide analysis, using publicly available single-nucleotide polymorphisms (SNPs) data sets, identified multiple genomic regions associated with variation in seed longevity. Reverse genetics of 20 candidate genes in Columbia ecotype resulted in seven genes positive for seed longevity (PSAD1, SSLEA, SSTPR, DHAR1, CYP86A8, MYB47 and SPCH) and five negative ones (RBOHD, RBOHE, RBOHF, KNAT7 and SEP3). In this uniform genetic background, natural and accelerated ageing methods provided similar results for seed-longevity in knock-out mutants. The NADPH oxidases (RBOHs), the dehydroascorbate reductase (DHAR1) and the photosystem I subunit (PSAD1) highlight the important role of oxidative stress on seed ageing. The cytochrome P-450 hydroxylase, CYP86A8, and the transcription factors, MYB47, KNAT7 and SEP3, support the protecting role of the seed coat during seed ageing.

An Arabidopsis Mutant Over-Expressing Subtilase SBT4.13 Uncovers the Role of Oxidative Stress in the Inhibition of Growth by Intracellular Acidification.

Intracellular acid stress inhibits plant growth by unknown mechanisms and it occurs in acidic soils and as consequence of other stresses. In order to identify mechanisms of acid toxicity, we screened activation-tagging lines of Arabidopsis thaliana for tolerance to intracellular acidification induced by organic acids. A dominant mutant, sbt4.13-1D, was isolated twice and shown to over-express subtilase SBT4.13, a protease secreted into endoplasmic reticulum. Activity measurements and immuno-detection indicate that the mutant contains less plasma membrane H+-ATPase (PMA) than wild type, explaining the small size, electrical depolarization and decreased cytosolic pH of the mutant but not organic acid tolerance. Addition of acetic acid to wild-type plantlets induces production of ROS (Reactive Oxygen Species) measured by dichlorodihydrofluorescein diacetate. Acid-induced ROS production is greatly decreased in sbt4.13-1D and atrboh-D,F mutants. The latter is deficient in two major NADPH oxidases (NOXs) and is tolerant to organic acids. These results suggest that intracellular acidification activates NOXs and the resulting oxidative stress is important for inhibition of growth. The inhibition of acid-activated NOXs in the sbt4.13-1D mutant compensates inhibition of PMA to increase acid tolerance.

PRX2 and PRX25, peroxidases regulated by COG1, are involved in seed longevity in Arabidopsis.

Permeability is a crucial trait that affects seed longevity and is regulated by different polymers including proanthocyanidins, suberin, cutin and lignin located in the seed coat. By testing mutants in suberin transport and biosynthesis, we demonstrate the importance of this biopolymer to cope with seed deterioration. Transcriptomic analysis of cog1-2D, a gain-of-function mutant with increased seed longevity, revealed the upregulation of several peroxidase genes. Reverse genetics analysing seed longevity uncovered redundancy within the seed coat peroxidase gene family; however, after controlled deterioration treatment, seeds from the prx2 prx25 double and prx2 prx25 prx71 triple mutant plants presented lower germination than wild-type plants. Transmission electron microscopy analysis of the seed coat of these mutants showed a thinner palisade layer, but no changes were observed in proanthocyanidin accumulation or in the cuticle layer. Spectrophotometric quantification of acetyl bromide-soluble lignin components indicated changes in the amount of total polyphenolics derived from suberin and/or lignin in the mutant seeds. Finally, the increased seed coat permeability to tetrazolium salts observed in the prx2 prx25 and prx2 prx25 prx71 mutant lines suggested that the lower permeability of the seed coats caused by altered polyphenolics is likely to be the main reason explaining their reduced seed longevity.

Eficacia de la creación de un programa de Escuela de pacientes en la detección de necesidades en pacientes con Poliquistosis Renal Autosómica Dominante / Efficacy of the creation of a patient school program in the detection of needs in patients with autosomal dominant polycystic kidney disease

Introducción: La Poliquistosis Renal Autosómica Dominante es una enfermedad renal crónica responsable del 10% de los casos de insuficiencia renal terminal. La participación y los grupos de apoyo entre iguales son herramientas que mejoran el bienestar, evitando complicaciones y retrasando el avance de la enfermedad. Objetivos: Detectar necesidades informativas, así como recursos de apoyo, en este grupo de pacientes mediante la puesta en marcha de una Escuela de Pacientes con poliquistosis renal autosómica dominante. Material y Método: Se utilizó un diseño mixto (cuantitativo y cualitativo). El estudio se desarrolló mediante cuatro fases: 1) Grupo focal: pacientes con poliquistosis renal y sus cuidadores; 2) Selección de los pacientes expertos; 3) Elaboración de los contenidos del programa de la Escuela de pacientes con poliquitstosis renal autosómica dominante; 4) Pilotaje del programa. Resultados: Se detectaron necesidades de información referentes al tratamiento oral y al afrontamiento de la poliquistosis renal que no están cubiertas por los equipos de nefrología. Conclusiones: La Escuela de Pacientes ha demostrado ser una herramienta útil para detectar necesidades y recursos en pacientes con poliquistosis renal autosómica dominante que han de enfrentarse a una enfermedad crónica donde se requiere la participación del paciente para garantizar la adhesión al tratamiento

Introduction: Autosomal Dominant Polycystic Kidney Disease is a chronic kidney disease responsible for 10% of cases of end-stage renal failure. Participation and peer support groups are tools that improve well-being, avoiding complications and delaying disease progression. Objectives: To detect information needs, as well as support resources, in patients with autosomal dominant polycystic kidney disease trough a Patient School. Material and Method: A mixed design (quantitative and qualitative) was used. The study was developed through four phases: 1) Focus group: patients with autosomal dominant polycystic kidney disease and their caregivers; 2) Selection of expert patients; 3) Preparation of the contents of the program of the Patient School with autosomal dominant polycystic kidney disease; 4) Piloting the program. Results: Information needs regarding oral treatment and coping with autosomal dominant polycystic kidney disease were detected, which are not covered by nephrology teams. Conclusions: Patients School has proven to be a useful tool to detect needs and resources in patients with autosomal dominant polycystic kidney disease who have to face a chronic disease where patient participation is required to ensure adherence to treatment

A vascular smooth muscle cell X-box binding protein 1 and transglutaminase 2 regulatory circuit limits neointimal hyperplasia.

Neointimal hyperplasia, stimulated by injury and certain vascular diseases, promotes artery obstruction and tissue ischemia. In vascular smooth muscle cell (VSMCs), multiple modulators of protein handling machinery regulate intimal hyperplasia. These include elements of the VSMC unfolded protein response to endoplasmic reticulum stress (UPRER), and transglutaminase 2 (TG2), which catalyzes post-translational protein modification. Previous results for deficiency of UPRER-specific mediator XBP1, and of TG2, have been significant, but in multiple instances contradictory, for effects on cultured VSMC function, and, using multiple models, for neointimal hyperplasia in vivo. Here, we engineered VSMC-specific deficiency of XBP1, and studied cultured VSMCs, and neointimal hyperplasia in response to carotid artery ligation in vivo. Intimal area almost doubled in Xbp1fl/fl SM22α-CRE+ mice 21 days post-ligation. Cultured murine Xbp1 deficient VSMCs migrated more in response to platelet derived growth factor (PDGF) than control VSMCs, and had an increased level of inositol-requiring enzyme 1α (Ire1α), a PDGF receptor-binding UPRER transmembrane endonuclease whose substrates include XBP1. Cultured XBP1-deficient VSMCs demonstrated decreased levels of TG2 protein, in association with increased TG2 polyubiquitination, but with increased TG transamidation catalytic activity. Moreover, IRE1α, and TG2-specific transamidation cross-links were increased in carotid artery neointima in Xbp1fl/fl SM22α-CRE+ mice. Cultured TG2-deficient VSMCs had decreased XBP1 associated with increased IRE1α, and increased migration in response to PDGF. Neointimal hyperplasia also was significantly increased in Tgm2fl/fl SM22α-CRE+ mice at 21 days after carotid ligation. In conclusion, a VSMC regulatory circuit between XBP1 and TG2 limits neointimal hyperplasia in response to carotid ligation.

Polyamines interfere with protein ubiquitylation and cause depletion of intracellular amino acids: a possible mechanism for cell growth inhibition.

Spermidine is a polyamine present in eukaryotes with essential functions in protein synthesis. At high concentrations spermidine and norspermidine inhibit growth by unknown mechanisms. Transcriptomic analysis of the effect of norspermidine on the plant Arabidopsis thaliana indicates upregulation of the response to heat stress and denatured proteins. Accordingly, these polyamines inhibit protein ubiquitylation, both in vivo (in yeast, Arabidopsis, and human Hela cells) and in vitro (with recombinant ubiquitin ligase). This interferes with protein degradation by the proteasome, a situation known to deplete cells of amino acids. Norspermidine treatment of yeast cells induces amino acid depletion, and supplementation of media with amino acids counteracts growth inhibition and cellular amino acid depletion but not inhibition of protein polyubiquitylation.

Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling.

BACKGROUND: Arhalofenate acid, the active acid form of arhalofenate, is a non-agonist peroxisome proliferator-activated receptor γ (PPARγ) ligand, with uricosuric activity via URAT1 inhibition. Phase II studies revealed decreased acute arthritis flares in arhalofenate-treated gout compared with allopurinol alone. Hence, we investigated the anti-inflammatory effects and mechanisms of arhalofenate and its active acid form for responses to monosodium urate (MSU) crystals. METHODS: We assessed in-vivo responses to MSU crystals in murine subcutaneous air pouches and in-vitro responses in murine bone marrow-derived macrophages (BMDMs) by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE/Western blot, immunostaining, and transmission electron microscopy analyses. RESULTS: Oral administration of arhalofenate (250 mg/kg) blunted total leukocyte ingress, neutrophil influx, and air pouch fluid interleukin (IL)-1ß, IL-6, and CXCL1 in response to MSU crystal injection (p < 0.05 for each). Arhalofenate acid (100 µM) attenuated MSU crystal-induced IL-1ß production in BMDMs via inhibition of NLRP3 inflammasome activation. In addition, arhalofenate acid dose-dependently increased activation (as assessed by phosphorylation) of AMP-activated protein kinase (AMPK). Studying AMPKα1 knockout mice, we elucidated that AMPK mediated the anti-inflammatory effects of arhalofenate acid. Moreover, arhalofenate acid attenuated the capacity of MSU crystals to suppress AMPK activity, regulated expression of multiple downstream AMPK targets that modulate mitochondrial function and oxidative stress, preserved intact mitochondrial cristae and volume density, and promoted anti-inflammatory autophagy flux in BMDMs. CONCLUSIONS: Arhalofenate acid is anti-inflammatory and acts via AMPK activation and its downstream signaling in macrophages. These effects likely contribute to a reduction of gout flares.