Source-to-sink partitioning is altered by changes in the expression of the transcription factor AtHB5 in Arabidopsis.

Carbohydrates are transported from source to sink tissues. The efficiency of this transport determines plant growth and development. The process is finely regulated and transcription factors are crucial in its modulation. AtHB5 is a homeodomain-leucine zipper I transcription factor that is repressed during stem maturation. However, its function in this developmental event is unknown. Here, we investigated the expression pattern and role of AtHB5. AtHB5 was expressed in roots, hypocotyls, stems, petioles, pedicels, and central leaf veins. athb5 mutant plants exhibited wider and more lignified stems than controls, whereas AtHB5 overexpressors showed the opposite phenotype. Cross sections of athb5 mutant stems showed enlarged vascular bundle, xylem, phloem, and petiole areas, whereas AtHB5 overexpressors had callose deposits. Several genes involved in starch biosynthesis and degradation had altered transcript levels in athb5 mutants and AtHB5 overexpressors. Rosette and stem biomass was enhanced in athb5 mutants, positively impacting seed yield, protein, and lipid content. Moreover, these effects were more evident in debranched plants. Finally, transport to roots was significantly slowed in AtHB5 overexpressors. Altogether, the results indicated that AtHB5 is a negative modulator of carbon partitioning and sucrose transport from source to sink tissues, and its overexpression diminished plant biomass and seed yield.

Monitoring Indicators of Universal Access to Accessible and Safe Anesthetic and Surgical Care in a Peruvian Region: An Ambispective Study.

INTRODUCTION: The Lancet Commission on Global Surgery indicators for monitoring anesthetic and surgical care allow the identification of access barriers, evaluate the safety of surgeries, facilitate planning, and assess changes over time. The primary objective was to measure these indicators in all health facilities of a Peruvian region in 2020. METHODS: This was an ambispective observational study to measure the anesthetic and surgical care indicators in Piura, a region in Peru, between January 2020 and June 2021. Public and private health facilities in the Piura region that performed surgical care or had specialists from any surgical specialty participated in the study. Data were collected from all regional health facilities that provided surgical care to estimate the density of surgical workforce. Likewise, the percentage of the population with access to an operating room within 2 h was estimated using georeferenced tools. Finally, a public database was accessed to determine the surgical volume, the percentage of the regional population protected with health insurance. RESULTS: In 2020, 88.4% of the inhabitants of this Peruvian region had access to timely essential surgery. There were 18.4 surgical specialists and 1174 surgeries per 100,000 populations, and 91% of the population had health insurance. In addition, there was a rate of 2.1 working operating rooms per 100,000 inhabitants in 2021. CONCLUSIONS: This Peruvian region presented an increasing trend with respect to the population's access to essential and timely surgical care, and health insurance coverage. However, the workforce distribution was inequitable among the provinces of the region, the surgical volume was reduced, and timely access was hindered because of the SARS-CoV-2 pandemic.

Comparative Proteomics of Potato Cultivars with a Variable Dormancy Period.

The control of the duration of the dormancy phase is a significant challenge in the potato industry and for seed producers. However, the proteome landscape involved in the regulation of the length of the dormancy period over potato cultivars remains largely unexplored. In this study, we performed for the first time a comparative proteome profiling of potato cultivars with differential duration of tuber dormancy. More specifically, the proteome profiling of Agata, Kennebec and Agria commercial potato varieties with short, medium and medium-long dormancy, respectively, was assessed at the endodormancy stage using high-resolution two-dimensional electrophoresis (2-DE) coupled to reversed-phase liquid chromatography-tandem mass spectrometry (LC-TripleTOF MS/MS). A total of 11 proteins/isoforms with statistically significant differential abundance among cultivars were detected on 2-DE gels and confidently identified by LC-TripleTOF MS/MS. Identified proteins have known functions related to tuber development, sprouting and the oxylipins biosynthesis pathway. Fructokinase, a mitochondrial ADP/ATP carrier, catalase isozyme 2 and heat shock 70 kDa were the proteins with the strongest response to dormancy variations. To the best of our knowledge, this study reports the first candidate proteins underlying variable dormancy length in potato cultivars.

Prevalence, Incidence, and Clinical-Epidemiological Characterization of Amyotrophic Lateral Sclerosis in Antioquia: Colombia.

INTRODUCTION: The prevalence and incidence of amyotrophic lateral sclerosis (ALS) is not fully established, and this varies depending on the studied population. OBJECTIVE: To estimate the prevalence/incidence of ALS patients in Antioquia-Colombia. METHODOLOGY: Observational/descriptive study by reviewing clinical records from 2010 to 2014. Cases with possible, probable and definite ALS were included. To estimate the prevalence/incidence, capture-recapture method was used. RESULTS: Point prevalence in December 2014 was 4.9/100,000 (95% CI 2.0-7.8), and the incidence was 1.4/100,000/year (95% CI 0.5-2.2). The median survival was 4 years. Spinal-onset was observed in 62.4% of the included patients. CONCLUSION: Prevalence, incidence and clinical presentation of ALS in Antioquia are similar to most studies reported worldwide. However, prevalence in Antioquia seems to be slightly higher than in other studies from Latin -American countries. This may derive from the inclusion criteria and case detection methodology adopted, but sociodemographic and genetic factors should be considered.

Evaluation of linseed oil oleogels to partially replace pork backfat in fermented sausages.

BACKGROUND: Nowadays, fat replacement in meat products is a matter of concern in the meat industry. The objective of this study was to evaluate the replacement of pork backfat with two oleogels of linseed in dry-cured sausages. RESULTS: Five batches of dry-cured sausages were prepared with two oleogels, a mixture of γ-oryzanol and ß-sitosterol (SO) and beeswax (B), at two levels of replacement (20% and 40%) (SO-20, SO-40, B-20, and B-40, respectively) and a control batch. The fatty acid profile improved in terms of nutrition: the polyunsaturated fatty acid / saturated fatty acid (PUFA/SFA) and n-6/n-3 ratio was about 1.41 and 0.93 for the higher levels of replacement, SO-40 and B-40, respectively. Quality parameters such as pH and color also changed with the inclusion of oleogels, resulting in changes in the sensory quality. CONCLUSION: Oleogels based on linseed enabled the replacement of pork backfat in fermented sausages. Depending on the level of fat substitution, such oleogels could replace fat in dry-cured sausages at the industrial level. © 2019 Society of Chemical Industry.

The first evidence of global meat phosphoproteome changes in response to pre-slaughter stress.

BACKGROUND: Pre-slaughter stress (PSS) impairs animal welfare and meat quality. Dark, firm and dry (DFD) are terms used to designate poor quality meats induced by PSS. Protein phosphorylation can be a potentially significant mechanism to explain rapid and multiple physiological and biochemical changes linked to PSS-dependent muscle-to-meat conversion. However, the role of reversible phosphorylation in the response to PSS is still little known. In this study, we report a comparative phosphoproteomic analysis of DFD and normal meats at 24 h post-mortem from the longissimus thoracis (LT) bovine muscle of male calves of the Rubia Gallega breed. For this purpose, two-dimensional gel electrophoresis (2-DE), in-gel multiplex identification of phosphoproteins with PRO-Q Diamond phosphoprotein-specific stain, tandem (MALDI-TOF/TOF) mass spectrometry (MS), novel quantitative phosphoproteomic statistics and bioinformatic tools were used. RESULTS: Noticeable and statistically significant differences in the extent of protein phosphorylation were detected between sample groups at the qualitative and quantitative levels. Overall phosphorylation rates across significantly changed phosphoproteins were about three times higher in DFD than in normal meat. Significantly changed phosphoproteins involved a variable number of isoforms of 13 myofibrillar and sarcoplasmic nonredundant proteins. However, fast skeletal myosin light chain 2 followed by troponin T, F-actin-capping and small heat shock proteins showed the greatest phosphorylation change, and therefore they were the most important phosphoproteins underlying LT muscle conversion to DFD meat in the Rubia Gallega breed. CONCLUSIONS: This is the first study reporting global meat phosphoproteome changes in response to PSS. The results show that reversible phosphorylation is a relevant mechanism underlying PSS response and downstream effects on meat quality. This research opens up novel horizons to unravel the complex molecular puzzle underlying muscle-to-meat conversion in response to PSS.

The Major Storage Protein in Potato Tuber Is Mobilized by a Mechanism Dependent on Its Phosphorylation Status.

The role of the protein phosphorylation mechanism in the mobilization of vegetative storage proteins (VSPs) is totally unknown. Patatin is the major VSP of the potato (Solanum tuberosum L.) tuber that encompasses multiple differentially phosphorylated isoforms. In this study, temporal changes in the phosphorylation status of patatin isoforms and their involvement in patatin mobilization are investigated using phosphoproteomic methods based on targeted two-dimensional electrophoresis (2-DE). High-resolution 2-DE profiles of patatin isoforms were obtained in four sequential tuber life cycle stages of Kennebec cultivar: endodormancy, bud break, sprouting and plant growth. In-gel multiplex identification of phosphorylated isoforms with Pro-Q Diamond phosphoprotein-specific stain revealed an increase in the number of phosphorylated isoforms after the tuber endodormancy stage. In addition, we found that the phosphorylation status of patatin isoforms significantly changed throughout the tuber life cycle (P < 0.05) using the chemical method of protein dephosphorylation with hydrogen fluoride-pyridine (HF-P) coupled to 2-DE. More specifically, patatin phosphorylation increased by 32% from endodormancy to the tuber sprouting stage and subsequently decreased together with patatin degradation. Patatin isoforms were not randomly mobilized because highly phosphorylated Kuras-isoforms were preferably degraded in comparison to less phosphorylated non-Kuras isoforms. These results lead us to conclude that patatin is mobilized by a mechanism dependent on the phosphorylation status of specific isoforms.

Role of Counterions in Molecularly Imprinted Polymers for Anionic Species.

Small-molecule oxoanions are often imprinted noncovalently as carboxylates into molecularly imprinted polymers (MIPs), requiring the use of an organic counterion. Popular species are either pentamethylpiperidine (PMP) as a protonatable cation or tetraalkylammonium (TXA) ions as permanent cations. The present work explores the influence of the TXA as a function of their alkyl chain length, from methyl to octyl, using UV/vis absorption, fluorescence titrations, and HPLC as well as MD simulations. Protected phenylalanines (Z-l/d-Phe) served as templates/analytes. While the influence of the counterion on the complex stability constants and anion-induced spectral changes shows a monotonous trend with increasing alkyl chain length at the prepolymerization stage, the cross-imprinting/rebinding studies showed a unique pattern that suggested the presence of adaptive cavities in the MIP matrix, related to the concept of induced fit of enzyme-substrate interaction. Larger cavities formed in the presence of larger counterions can take up pairs of Z-x-Phe and smaller TXA, eventually escaping spectroscopic detection. Correlation of the experimental data with the MD simulations revealed that counterion mobility, the relative distances between the three partners, and the hydrogen bond lifetimes are more decisive for the response features observed than actual distances between interacting atoms in a complex or the orientation of binding moieties. TBA has been found to yield the highest imprinting factor, also showing a unique dual behavior regarding the interaction with template and fluorescent monomer. Finally, interesting differences between both enantiomers have been observed in both theory and experiment, suggesting true control of enantioselectivity. The contribution concludes with suggestions for translating the findings into actual MIP development.

Label-Free Quantification of Anti-TNF-α in Patients Treated with Adalimumab Using an Optical Biosensor.

This study describes the development of an immunosensory label-free quantification methodology based on surface plasmon resonance (SPR) and its applicability in measuring/evaluating therapeutic drug monitoring (TDM) of anti-TNF-α monoclonal antibody (adalimumab) in rheumatoid arthritis (RA) patients. The experimental parameters evaluated in this study were immobilising ligands by pre-concentration assays, sensor surface regeneration, ascertaining the method's sensitivity and correlating the results from quantifying plasma samples by ELISA immunoassay. The results showed that TNF-α quantification values (in RU) were significantly different when comparing patients (~50-250 RU) to controls (~10-20 RU). Likewise, there was 0.97 correlation for patients and 0.91 for healthy volunteers using SPR and ELISA comparison methodologies. SPR immunosensory detection provided a precise, sensitive strategy, along with real-time determination, for quantifying adalimumab, having great potential for clinical routine regarding TDM.

Advances in the Biology of Seed and Vegetative Storage Proteins Based on Two-Dimensional Electrophoresis Coupled to Mass Spectrometry.

Seed storage proteins play a fundamental role in plant reproduction and human nutrition. They accumulate during seed development as reserve material for germination and seedling growth and are a major source of dietary protein for human consumption. Storage proteins encompass multiple isoforms encoded by multi-gene families that undergo abundant glycosylations and phosphorylations. Two-dimensional electrophoresis (2-DE) is a proteomic tool especially suitable for the characterization of storage proteins because of their peculiar characteristics. In particular, storage proteins are soluble multimeric proteins highly represented in the seed proteome that contain polypeptides of molecular mass between 10 and 130 kDa. In addition, high-resolution profiles can be achieved by applying targeted 2-DE protocols. 2-DE coupled with mass spectrometry (MS) has traditionally been the methodology of choice in numerous studies on the biology of storage proteins in a wide diversity of plants. 2-DE-based reference maps have decisively contributed to the current state of our knowledge about storage proteins in multiple key aspects, including identification of isoforms and quantification of their relative abundance, identification of phosphorylated isoforms and assessment of their phosphorylation status, and dynamic changes of isoforms during seed development and germination both qualitatively and quantitatively. These advances have translated into relevant information about meaningful traits in seed breeding such as protein quality, longevity, gluten and allergen content, stress response and antifungal, antibacterial, and insect susceptibility. This review addresses progress on the biology of storage proteins and application areas in seed breeding using 2-DE-based maps.

Evidence for phosphorylation of the major seed storage protein of the common bean and its phosphorylation-dependent degradation during germination.

Phaseolin is the major seed storage protein of common bean, Phaseolus vulgaris L., accounting for up to 50 % of the total seed proteome. The regulatory mechanisms responsible for the synthesis, accumulation and degradation of phaseolin in the common bean seed are not yet sufficiently known. Here, we report on a systematic study in dormant and 4-day germinating bean seeds from cultivars Sanilac (S) and Tendergreen (T) to explore the presence and dynamics of phosphorylated phaseolin isoforms. High-resolution two-dimensional electrophoresis in combination with the phosphoprotein-specific Pro-Q Diamond phosphoprotein fluorescent stain and chemical dephosphorylation by hydrogen fluoride-pyridine enabled us to identify differentially phosphorylated phaseolin polypeptides in dormant and germinating seeds from cultivars S and T. Phosphorylated forms of the two subunits of type α and ß that compose the phaseolin were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and MALDI-TOF/TOF tandem MS. In addition, we found that the levels of phosphorylation of the phaseolin changed remarkably in the seed transition from dormancy to early germination stage. Temporal changes in the extent of phosphorylation in response to physiological and metabolic variations suggest that phosphorylated phaseolin isoforms have functional significance. In particular, this prospective study supports the hypothesis that mobilization of the phaseolin in germinating seeds occurs through the degradation of highly phosphorylated isoforms. Taken together, our results indicate that post-translational phaseolin modifications through phosphorylations need to be taken into consideration for a better understanding of the molecular mechanisms underlying its regulation.

Simultaneous Bilateral Quadriceps Tendon Rupture Secondary to Parathyroid Carcinoma: A Case Report.

Simultaneous bilateral quadriceps tendon ruptures are a rare occurrence commonly associated with a traumatic event or systemic disease. A 31-year-old man presented with simultaneous bilateral quadriceps tendon ruptures with associated hyperparathyroidism secondary to parathyroid carcinoma. The injury occurred after the patient attempted to lift a small wooden log from the ground. We discussed the multidisciplinary management of this patient resulting in bilateral quadriceps tendon repairs, tumor resection, and oncological and endocrinological restoration. Clinical follow-up is reported at 15 years after surgery. Parathyroid carcinoma is an extremely rare cancer and rarely the cause of hyperparathyroidism. The systemic effects of the tumor eventually lead to the rupturing of both quadriceps tendons. Orthopaedic physicians must remain vigilant in identifying the root cause of injuries that are atypical in nature.

Nonlinear control with friction compensation to swing-up a Furuta pendulum.

Different works in literature have reported that nonlinear controllers based on the energy approach are not effective to completely swing-up an inverted pendulum subjected to friction. Most studies trying to solve this issue consider static friction models in the design of controllers. This consideration is mainly because the stability proof of the system with dynamic friction in closed-loop is difficult. Hence, this paper presents a nonlinear controller with friction compensation to swing-up a Furuta pendulum with dynamic friction. With this aim, we consider that only the active joint of the system is subjected to friction, which is represented via a dynamic model, namely, the Dahl model. We first present Furuta Pendulum dynamic model with dynamic friction. Then, by slightly modifying an energy-based controller that has been previously reported in literature and by including friction compensation, we propose a nonlinear controller that allows to swing-up completely a Furuta pendulum subjected to friction. The unmeasurable friction state is estimated through a nonlinear observer and a stability analysis of the closed-loop system is accomplished with the direct Lyapunov method. Finally, successful experimental results are presented for a Furuta pendulum prototype built by authors. This shows the effectiveness of the proposed controller in achieving a complete swing-up of the Furuta pendulum, in a time feasible for experimental implementation, and ensuring closed-loop stability.

Surgical care and trauma patients capacity in Piura, Perú - Cross-sectional study.

Introduction: Low and medium income countries face challenges in access and delivery of surgical care, resulting in a high number of deaths and disabled individuals. Objective: To estimate the capacity to provide surgical and trauma care in public hospitals in the Piura region, Perú, a middle income country. Methods: A survey was administered in public hospitals in the Peruvian region of Piura, which combined the Spanish versions of the PIPES and INTACT surveys, and the WHO situational analysis tool. The extent of the event was assessed based in the absolute differences between the medians of the scores estimated, and the Mann-Whitney bilateral tests, according to the geographical location and the level of hospital complexity. Results: Seven public hospitals that perform surgeries in the Piura region were assessed. Three provinces (3/8) did not have any complexity healthcare institutions. The average hospital in the peripheral provinces tended to be smaller than in the capital province in INTACT (8.25 vs. 9.5, p = 0.04). Additionally, water supply issues were identified (2/7), lack of incinerator (3/7), lack of uninterrupted availability of a CT-scanner (5/7) and problems with working hours; in other words, the blood banks in two hospitals were not open 24 hours. Conclusions: There is a significant inequality among the provinces in the region in terms of their trauma care capacities and several shortfalls in the public sector healthcare infrastructure. This information is required to conduct future research on capacity measurements in every public and private institution in the Peruvian region of Piura.

Introducción: Los países de ingresos bajos y medianos tienen problemas en el acceso y la provisión de atención quirúrgica, lo cual ocasiona un alto número de fallecimientos y de personas con discapacidad. Objetivo: Estimar la capacidad para la atención quirúrgica y de pacientes traumatizados en los hospitales públicos en la región de Piura, Perú, un país de ingreso mediano. Métodos: En los hospitales públicos de la región peruana de Piura se aplicó una encuesta que combinaba las versiones en español de las encuestas PIPES e INTACT y de la herramienta de análisis situacional de la Organizacion Mundial de la Salud (OMS). Se evaluó la magnitud del evento mediante las diferencias absolutas entre las medianas de los puntajes calculados y pruebas bilaterales de Mann-Whitney según la ubicación geográfica y el nivel de complejidad hospitalaria. Resultados: Se evaluaron siete hospitales públicos que realizan cirugía en la región de Piura. Tres provincias (3/8) no contaban con instituciones sanitarias con complejidad de hospital. La mediana de los hospitales de las provincias periféricas tuvo tendencia a ser menor que la de la provincia capital en la INTACT (8,25 vs. 9,5, p = 0,04). Asimismo, se hallaron problemas de abastecimiento de agua (2/7), ausencia de incinerador (3/7), falta de funcionamiento permanente de tomógrafo (5/7) y problemas con el horario de funcionamiento de los bancos de sangre, ya que no funcionaban las 24 horas del día en 2 hospitales (2/7). Conclusiones: Se describe la alta desigualdad entre las provincias de la región en la capacidad de atención de trauma y varias carencias en la infraestructura sanitaria del sector público. Esta información es necesaria para desarrollar futura investigación de medición de capacidades en todos los establecimientos públicos y privados de la región peruana de Piura.

On the uses and applications of the most commonly used measures of linkage disequilibrium from the comparative analysis of their statistical properties.

BACKGROUND/OBJECTIVE: The analysis of linkage disequilibrium is relevant for the exploration of the structure and evolution of genomes and for the gene mapping of quantitative characters and human diseases. The strength of linkage disequilibrium between diallelic loci is commonly measured by the coefficients D' and r. Recent studies suggest that r is more useful than D' as a general measure of the strength of disequilibrium because it provides much more precise (lower sampling variance) and accurate (lower bias) estimates of disequilibrium. We compared for the first time the statistical properties of D' and r taking into account their differences in range. METHODS: The sampling properties of D' and r were evaluated by simulation under a variety of realistic population conditions and varying sample sizes using standardised statistics that allow for comparisons of the precision, accuracy and efficiency of estimates with different ranges. RESULTS: Simulations revealed that estimates of r do not tend to be significantly more precise, accurate or efficient than those of D' when compared by means of standardised statistics. CONCLUSION: The supposed advantage of r over D' based on direct comparisons of their sampling distributions is more apparent than real. The obtained results are useful to assess the uses and applications of these widely used disequilibrium measures.

Phosphoproteome Analysis Using Two-Dimensional Electrophoresis Coupled with Chemical Dephosphorylation.

Protein phosphorylation is a reversible post-translational modification (PTM) with major regulatory roles in many cellular processes. However, the analysis of phosphoproteins remains the most challenging barrier in the prevailing proteome research. Recent technological advances in two-dimensional electrophoresis (2-DE) coupled to mass spectrometry (MS) have enabled the identification, characterization, and quantification of protein phosphorylation on a global scale. Most research on phosphoproteins with 2-DE has been conducted using phosphostains. Nevertheless, low-abundant and low-phosphorylated phosphoproteins are not necessarily detected using phosphostains and/or MS. In this study, we report a comparative analysis of 2-DE phosphoproteome profiles using Pro-Q Diamond phosphoprotein stain (Pro-Q DPS) and chemical dephosphorylation of proteins with HF-P from longissimus thoracis (LT) muscle samples of the Rubia Gallega cattle breed. We found statistically significant differences in the number of identified phosphoproteins between methods. More specifically, we found a three-fold increase in phosphoprotein detection with the HF-P method. Unlike Pro-Q DPS, phosphoprotein spots with low volume and phosphorylation rate were identified by HF-P technique. This is the first approach to assess meat phosphoproteome maps using HF-P at a global scale. The results open a new window for 2-DE gel-based phosphoproteome analysis.

Kidney Disease after Heart and Lung Transplantation.

Chronic kidney disease (CKD) is not only common after lung and heart transplantation but also is associated with increased morbidity and mortality due to multiple pre-, peri- and post-transplant factors. While the exact incidence of CKD in this population is not well-defined, it seems to have gradually increased over the years as older recipients are more frequently considered. The increasing success of the procedure and expanding transplant candidate pool has allowed many with comorbid conditions to receive a transplant, which was considered prohibitive in the past. This review presents risk factors that have been linked to CKD as well as interventions that may help alleviate this serious problem. The impact of pretransplant renal function and the overexaggerated role of chronic nephrotoxicity of calcineurin inhibitors is discussed in detail. Until the exact pathophysiology of kidney disease is better understood, there is a dire need to expand the research agenda beyond observational studies.