Machine learning guided rational design of a non-heme iron-based lysine dioxygenase improves its total turnover number.

Highly selective C-H functionalization remains an ongoing challenge in organic synthetic methodologies. Biocatalysts are robust tools for achieving these difficult chemical transformations. Biocatalyst engineering has often required directed evolution or structure-based rational design campaigns to improve their activities. In recent years, machine learning has been integrated into these workflows to improve the discovery of beneficial enzyme variants. In this work, we combine a structure-based machine-learning algorithm with classical molecular dynamics simulations to down select mutations for rational design of a non-heme iron-dependent lysine dioxygenase, LDO. This approach consistently resulted in functional LDO mutants and circumvents the need for extensive study of mutational activity before-hand. Our rationally designed single mutants purified with up to 2-fold higher yields than WT and displayed higher total turnover numbers (TTN). Combining five such single mutations into a pentamutant variant, LPNYI LDO, leads to a 40% improvement in the TTN (218±3) as compared to WT LDO (TTN = 160±2). Overall, this work offers a low-barrier approach for those seeking to synergize machine learning algorithms with pre-existing protein engineering strategies.

Biochemical and structural insights of a recombinant AA16 LPMO from the marine and sponge-symbiont Peniophora sp.

Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that oxidize polysaccharides, leading to their cleavage. LPMOs are classified into eight CAZy families (AA9-11, AA13-17), with the functionality of AA16 being poorly characterized. This study presents biochemical and structural data for an AA16 LPMO (PnAA16) from the marine sponge symbiont Peniophora sp. Phylogenetic analysis revealed that PnAA16 clusters separately from previously characterized AA16s. However, the structural modelling of PnAA16 showed the characteristic immunoglobulin-like fold of LPMOs, with a conserved his-brace motif coordinating a copper ion. The copper-bound PnAA16 showed greater thermal stability than its apo-form, highlighting copper's role in enzyme stability. Functionally, PnAA16 demonstrated oxidase activity, producing 5 µM H2O2 after 30 min, but showed 20 times lower peroxidase activity (0.27 U/g) compared to a fungal AA9. Specific activity assays indicated that PnAA16 acts only on cellohexaose, generating native celloligosaccharides (C3 to C5) and oxidized products with regioselective oxidation at C1 and C4 positions. Finally, PnAA16 boosted the activity of a cellulolytic cocktail for cellulose saccharification in the presence of ascorbic acid, hydrogen peroxide, or both. In conclusion, the present work provides insights into the AA16 family, expanding the understanding of their structural and functional relationships and biotechnological potential.

Dataset on sub-daily vertical profiles of physicochemical parameters and chlorophyll concentration in El Val reservoir, together with its daily meteorological data, storage state and downstream flow (2018-2022).

The dataset addressed in this article contains parameters about El Val reservoir (province of Zaragoza, Spain). It includes physicochemical variables, the water level, the stored water volume, its meteorological conditions and the flow rate of its effluent, the Queiles River, a few metres downstream of the dam. The El Val reservoir stores water from the Val River, but it also receives water from the Queiles River through a pipeline and from several ravines. The dam releases on the Queiles River, which is a tributary of the Ebro River (the second one in Spain in length and discharge rate). A multiparametric probe (aquaDam, Adasa Systems), hanging from a structure located in the dam, every 6 h makes a vertical profile taking the measurements at each metre of depth from the surface to approximately 573 m above sea level (m.a.s.l.), in other words, between 2 and 3 m above the bottom outlet. This probe collects data of water temperature, pH, ORP, conductivity, dissolved oxygen, turbidity and chlorophyll concentration. Meteorological data are collected in the nearest weather station, located in the municipality of Los Fayos which is about 500 m downstream of the dam. These include daily accumulated precipitation, daily maximum and average solar irradiance, daily maximum, minimum and average air temperature and daily average wind speed. The water level and volume of stored water and the flow rate of the Queiles River are collected in the El Val monitoring station and the Queiles River gauge station respectively, and are also provided on a daily basis. These data are useful to feed deterministic, data driven or hybrid hydrological models with different purposes, like the identification of the impact of meteorological conditions on the physicochemical properties of the reservoir as well as the assessment of different management strategies in the reservoir. This is a data article that additionally supports the work published in Ecological Informatics [1] where the use of common and readily available open data is promoted through its use to feed data driven models, in particular to infer the depth of the thermocline in reservoirs that are periodically or permanently thermally stratified. In that article a dataset derived from the one presented in this article is used.

Kidney transplant recipient with history of HIV, HBV, and past HCV infection.

Chronic viral infections caused by the human immunodeficiency virus (HIV), hepatitis C (HCV), and hepatitis B (HBV) are common among patients with end-stage renal disease (ESKD). These infections were once considered contraindications to kidney transplantation due to potential risks associated with long-term immunosuppression. Improved management and antiviral therapies have changed the prognosis and survival of this group of patients, along with an increased experience in transplanting people with these viral infections. We report the first successful kidney transplant in an ESKD patient on hemodialysis with a history of concomitant HIV, HCV and HBV infection in Mexico.

Stability Oracle: a structure-based graph-transformer framework for identifying stabilizing mutations.

Engineering stabilized proteins is a fundamental challenge in the development of industrial and pharmaceutical biotechnologies. We present Stability Oracle: a structure-based graph-transformer framework that achieves SOTA performance on accurately identifying thermodynamically stabilizing mutations. Our framework introduces several innovations to overcome well-known challenges in data scarcity and bias, generalization, and computation time, such as: Thermodynamic Permutations for data augmentation, structural amino acid embeddings to model a mutation with a single structure, a protein structure-specific attention-bias mechanism that makes transformers a viable alternative to graph neural networks. We provide training/test splits that mitigate data leakage and ensure proper model evaluation. Furthermore, to examine our data engineering contributions, we fine-tune ESM2 representations (Prostata-IFML) and achieve SOTA for sequence-based models. Notably, Stability Oracle outperforms Prostata-IFML even though it was pretrained on 2000X less proteins and has 548X less parameters. Our framework establishes a path for fine-tuning structure-based transformers to virtually any phenotype, a necessary task for accelerating the development of protein-based biotechnologies.

The hepatocyte epidermal growth factor receptor (EGFR) pathway regulates the cellular interactome within the liver fibrotic niche.

Liver fibrosis is the consequence of chronic liver injury in the presence of an inflammatory component. Although the main executors of this activation are known, the mechanisms that lead to the inflammatory process that mediates the production of pro-fibrotic factors are not well characterized. Epidermal growth factor receptor (EGFR) signaling in hepatocytes is essential for the regenerative processes of the liver; however, its potential role in regulating the fibrotic niche is not yet clear. Our group generated a mouse model that expresses an inactive truncated form of the EGFR specifically in hepatocytes (ΔEGFR mice). Here, we have analyzed the response of WT and ΔEGFR mice to chronic treatment with carbon tetrachloride (CCl4), which induces a pro-inflammatory and fibrotic process in the liver. The results indicated that the hallmarks of liver fibrosis were attenuated in CCl4-treated ΔEGFR mice when compared with CCl4-treated WT mice, coinciding with a faster resolution of the fibrotic process and ameliorated damage. The absence of EGFR activity in hepatocytes induced changes in the pattern of immune cells in the liver, with a notable increase in the population of M2 macrophages, more related to fibrosis resolution, as well as in the population of lymphocytes related to eradication of the damage. Transcriptome analysis of hepatocytes, and secretome studies of extracellular media from in vitro experiments, allowed us to elucidate the specific molecular mechanisms regulated by EGFR that mediate hepatocyte production of both pro-fibrotic and pro-inflammatory mediators; these have consequences for the deposition of extracellular matrix proteins, as well as for the immune microenvironment. Overall, our study uncovered novel mechanistic insights regarding EGFR kinase-dependent actions in hepatocytes that reveal its key role in chronic liver damage. © 2024 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Use of cannabidiol in the treatment of drug-refractory epilepsy in children and young adults: A systematic review.

Objectives: Epilepsy poses a significant challenge in pediatric and adolescent populations, impacting not only seizures but also psychological and cognitive comorbidities, leading to higher mortality rates than the general population. Drug-refractory epilepsy, resistant to conventional treatments, affects a range of 7-20% of pediatric patients. The search for alternative therapies has led to exploring the therapeutic potential of Cannabis sativa L. compounds, particularly cannabidiol (CBD). Examine the use of CBD for treating drug-refractory epilepsy in children and young adults, summarizing existing evidence on its efficacy. Materials and Methods: A systematic review, following Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, assessed studies from 2018 to 2023, focusing on CBD's efficacy and safety for treatment-resistant epilepsy in pediatric and juvenile populations. The search spanned seven databases, and the studies underwent rigorous screening and data extraction. Results: Out of 6351 identified articles, eight were selected for review. The included studies reported positive outcomes, with CBD leading to a reduction in seizure frequency ranging from 50% to complete seizure freedom. Adverse effects were mostly mild and reversible, including drowsiness, diarrhea, and loss of appetite. Conclusion: The CBD emerges as a promising tool for refractory epilepsy in pediatric patients, showing efficacy in reducing seizure frequency and improving overall quality of life. Despite mild and reversible adverse effects, CBD's benefits outweigh the risks. However, more research on long-term effects is needed to fully understand its implications.

The global prevalence and associated risk factors of Eimeria infection in domestic chickens: A systematic review and meta-analysis.

BACKGROUND: Eimeria is a protozoan parasite that affects poultry, particularly chickens, causing a disease known as coccidiosis. This disease imposes substantial significant economic challenges to the poultry sector. OBJECTIVES: The current study aimed to estimate the global prevalence and associated risk factors of Eimeria in domestic chickens. METHODS: Multiple databases (Scopus, PubMed, ProQuest, Web of Science and Google Scholar) were searched for articles published until June 2023. The pooled prevalence was estimated using a random-effects model with a 95% confidence interval. The statistical analysis was conducted using meta packages in R version (3.6.1). RESULTS: In total, 41 articles fulfilled the eligibility criteria. The global pooled prevalence was 44.3% (36.9%-51.8%) with Eimeria tenella (38.7%, 30.1%-47.7%) as the most prevalent species. The highest pooled prevalence was related to the Western Pacific Region (80.5%, 72.6%-87.3%) and urban areas (44.4%, 36.5%-52.6%). Moreover, areas with humid subtropical climates represent the highest overall prevalence (75.8%, 46.6%-95.9%). CONCLUSION: The necessity for robust and innovative strategies for preventing and managing this disease cannot be overstated. Addressing Eimeria impact is crucial not only for safeguarding poultry health but also for sustaining the economic viability of the poultry industry.

Aspectos a considerar para el diagnóstico del queratocono infantil / Aspects to consider for the diagnosis of infant keratoconus

Introducción: Globalmente, existe un aumento de la prevalencia del queratocono y su diagnóstico en edades tempranas. Se notifican un gran número de casos subclínicos y otros con una rápida progresión, condicionada por el inicio precoz de la enfermedad y la asociación a factores de riesgo. Objetivo: Describir los aspectos epidemiológicos, clínicos y el resultado de los medios de diagnóstico implicados en la detección precoz del queratocono infantil. Desarrollo: En niños con ametropía hay elementos que alertan la presencia de un queratocono como causa del defecto refractivo. Desde el punto de vista epidemiológico se encuentran: distribución geográfica, rol de la herencia y factores ambientales. Clínicamente se señalan los antecedentes de enfermedades, tales como las alergias, la presencia de miopía o astigmatismo miópico con inestabilidad refractiva y los signos clínicos relacionados con la progresión del cono. En los pacientes de riesgo es preciso realizar exámenes mediante diferentes medios de diagnóstico según su disponibilidad, siendo primordial el análisis refractivo, queratométrico y topográfico. Conclusiones: En la evaluación de los niños con ametropía se deben tener en cuenta elementos epidemiológicos y clínicos que permiten sospechar y diagnosticar precozmente el queratocono. En la interpretación de los resultados de los medios de diagnóstico involucrados en su detección, se deben considerar los hallazgos más frecuentes en la población infantil según el grado de progresión de la ectasia.

Introduction: Globally, there is an increase of the keratoconus prevalence and its diagnosis in early ages. A great number of subclinical cases and others with a quick progression are notified, conditioned by the early onset of the disease and the association with risk factors. Objective: To describe the epidemiological, clinical aspects and the result of diagnostic means involved in the early detection of infant keratoconus. Development: There are elements that alert the presence of a keratoconus as a cause of the refractive defect in children with ametropia. From the epidemiologic point of view they are: geographical distribution, heredity role and environmental factors. History of previous diseases are clinically pointed out, such as allergies, myopia or myopic astigmatism with refractive instability and the clinical signs related to cone progression. In risk patients it is necessary to carry out exams by means of different diagnostic means according to their availability, being essential the refractive, keratometric and topographic analysis. Conclusions: In the evaluation of children with ametropia, epidemiological and clinical elements should be taken into account that allow to suspect and early diagnose the keratoconus. In the interpretation of results of the diagnostic means involved in their detection the most frequent findings in the infant population, should be considered according to the ectasia degree of progression.

Asymmetric Synthesis of α-Chloroamides via Photoenzymatic Hydroalkylation of Olefins.

Photoenzymatic intermolecular hydroalkylations of olefins are highly enantioselective for chiral centers formed during radical termination but poorly selective for centers set in the C-C bond-forming event. Here, we report the evolution of a flavin-dependent "ene"-reductase to catalyze the coupling of α,α-dichloroamides with alkenes to afford α-chloroamides in good yield with excellent chemo- and stereoselectivity. These products can serve as linchpins in the synthesis of pharmaceutically valuable motifs. Mechanistic studies indicate that radical formation occurs by exciting a charge-transfer complex templated by the protein. Precise control over the orientation of molecules within the charge-transfer complex potentially accounts for the observed stereoselectivity. The work expands the types of motifs that can be prepared using photoenzymatic catalysis.

Biosensor and machine learning-aided engineering of an amaryllidaceae enzyme.

A major challenge to achieving industry-scale biomanufacturing of therapeutic alkaloids is the slow process of biocatalyst engineering. Amaryllidaceae alkaloids, such as the Alzheimer's medication galantamine, are complex plant secondary metabolites with recognized therapeutic value. Due to their difficult synthesis they are regularly sourced by extraction and purification from the low-yielding daffodil Narcissus pseudonarcissus. Here, we propose an efficient biosensor-machine learning technology stack for biocatalyst development, which we apply to engineer an Amaryllidaceae enzyme in Escherichia coli. Directed evolution is used to develop a highly sensitive (EC50 = 20 µM) and specific biosensor for the key Amaryllidaceae alkaloid branchpoint 4'-O-methylnorbelladine. A structure-based residual neural network (MutComputeX) is subsequently developed and used to generate activity-enriched variants of a plant methyltransferase, which are rapidly screened with the biosensor. Functional enzyme variants are identified that yield a 60% improvement in product titer, 2-fold higher catalytic activity, and 3-fold lower off-product regioisomer formation. A solved crystal structure elucidates the mechanism behind key beneficial mutations.

Development and Characterization of Electrodes Coated with Plasma-Synthesized Polypyrrole Doped with Iodine, Implanted in the Rat Brain Subthalamic Nucleus.

Biological treatments involve the application of metallic material coatings to enhance biocompatibility and properties. In invasive therapies, metallic electrodes are utilized, which are implanted in patients. One of these invasive therapeutic procedures is deep brain stimulation (DBS), an effective therapy for addressing the motor disorders observed in patients with Parkinson's disease (PD). This therapy involves the implantation of electrodes (IEs) into the subthalamic nucleus (STN). However, there is still a need for the optimization of these electrodes. Plasma-synthesized polypyrrole doped with iodine (PPPy/I) has been reported as a biocompatible and anti-inflammatory biomaterial that promotes nervous system regeneration. Given this information, the objective of the present study was to develop and characterize a PPPy/I-coated electrode for implantation into the STN. The characterization results indicate a uniform coating along the electrode, and physical-chemical characterization studies were conducted on the polymer. Subsequently, the IEs, both coated and uncoated with PPPy/I, were implanted into the STN of male rats of the Wistar strain to conduct an electrographic recording (EG-R) study. The results demonstrate that the IE coated with PPPy/I exhibited superior power and frequency signals over time compared to the uncoated IE (p < 0.05). Based on these findings, we conclude that an IE coated with PPPy/I has optimized functional performance, with enhanced integrity and superior signal quality compared to an uncoated IE. Therefore, we consider this a promising technological development that could significantly improve functional outcomes for patients undergoing invasive brain therapies.

Immunization of cattle with a Rhipicephalus microplus chitinase peptide containing predicted B-cell epitopes reduces tick biological fitness.

Rhipicephalus microplus, the cattle fever tick, is the most important ectoparasite impacting the livestock industry worldwide. Overreliance on chemical treatments for tick control has led to the emergence of acaricide-resistant ticks and environmental contamination. An immunological strategy based on vaccines offers an alternative approach to tick control. To develop novel tick vaccines, it is crucial to identify and evaluate antigens capable of generating protection in cattle. Chitinases are enzymes that degrade older chitin at the time of moulting, therefore allowing interstadial metamorphosis. In this study, 1 R. microplus chitinase was identified and its capacity to reduce fitness in ticks fed on immunized cattle was evaluated. First, the predicted amino acid sequence was determined in 4 isolates and their similarity was analysed by bioinformatics. Four peptides containing predicted B-cell epitopes were designed. The immunogenicity of each peptide was assessed by inoculating 2 cattle, 4 times at 21 days intervals, and the antibody response was verified by indirect ELISA. A challenge experiment was conducted with those peptides that were immunogenic. The chitinase gene was successfully amplified and sequenced, enabling comparison with reference strains. Notably, a 99.32% identity and 99.84% similarity were ascertained among the sequences. Furthermore, native protein recognition was demonstrated through western blot assays. Chitinase peptide 3 reduced the weight and oviposition of engorged ticks, as well as larvae viability, exhibiting a 71% efficacy. Therefore, chitinase 3 emerges as a viable vaccine candidate, holding promise for its integration into a multiantigenic vaccine against R. microplus.

Childhood obesity and its impact on the characteristics of gait stance phases: a cross-sectional study.

This study aimed to investigate the variations in foot type, laxity, dynamic characteristics of gait, and the characteristics of the stance phase of gait, in relation to body mass index (BMI) and groups of children of different ages. Additionally, it aimed to explore the correlations between BMI and these variables across children groups of different ages. A cross-sectional study was conducted involving 196 infants aged between 5 and 10 years old. The variables assessed included BMI, foot type, laxity, dynamic variables, and characteristics of the stance phase of gait. Significant variations were observed in foot type, laxity, certain dynamic variables, and characteristics of the stance phase of gait between normoweight (NW) and overweight/obese (OW/OB) groups among children aged between 5 and 10 years old (p ranged between 0.019 and 0.050). Moreover, BMI was also positively associated with the initial forefoot contact, heel off, total duration of the step, and forefoot contact phase of children 7 to 10 years of age (p ranged between < 0.010 and 0.040).   Conclusion: Children who are OW/OB had alterations at different stages of gait. Being OW/OB is related to alterations of the phases of gait mainly from 7 to 10 years of age, and spending more time in each of the phases of walking. This could indicate that children who are OW/OB, in addition to walking slower, overload the musculoskeletal system, subjecting their joints and muscles to greater stress. What is Known: • Children who are overweight (OW) and obese (OB) can experience changes in their musculoskeletal systems, posture, and gait due to increased body mass index. • OW and OB children experience additional stress on their musculoskeletal systems, impacting posture, biomechanics, mobility, physical activity, and daily tasks. Excessive plantar loading is linked to foot pain in adults. What is New: • Body mass index was positively associated with initial forefoot contact, heel off, total duration of the step, and forefoot contact phase in children aged 7 to 10 years old. OW/OB children aged 5-6 exhibited less ankle dorsiflexion and smaller relaxed calcaneal stance position (RCSP) as compared to normal-weight children. • Obese children aged 5-6 showed less pronation excursion, suggesting altered frontal plane movement due to RCSP differences. Children aged 7-8 who are OW/OB spent more time in certain gait phases, particularly in the forefoot contact phase. Being OW/OB is linked to altered gait parameters such as initial forefoot, heel off, total step duration, and forefoot contact phase. Being OW/OB was associated with a longer forefoot contact phase, particularly in the right foot.

Asimetría interocular en el queratocono pediátrico / Interocular Asymmetry in Pediatric Keratoconus

El queratocono es una ectasia corneal bilateral asimétrica, en etapas iniciales no se evidencia la afectación binocular. El conocimiento del grado de asimetría en el queratocono pediátrico permite tomar decisiones oportunas en el manejo de los pacientes. Se realizó este trabajo con el objetivo de describir las características clínicas y el tratamiento de tres casos pediátricos con diferente grado de asimetría interocular del queratocono, atendidos en la consulta provincial de ectasias corneales pediátricas en Ciego de Ávila. Los pacientes mostraron diferencia entre ambos ojos de los signos clínicos, la refracción y las variables topográficas. Al paciente dos se le diagnosticó queratocono en un ojo, sin evidencias clínicas ni topográficas de la enfermedad en el ojo contralateral. Todos los pacientes mostraron astigmatismo miópico compuesto en ambos ojos y se les indicó corrección óptica. Al paciente uno se le corrigió con cristales, al dos con lentes de contacto rígidos de gas permeable y al tercero con piggyback en un ojo y lentes de contacto rígidos de gas permeable en el otro. Presentaron, además, una ambliopía asociada y se les orientó tratamiento oclusivo y tareas de visión cercana en los pacientes dos y tres. Es frecuente encontrar asimetría interocular en pacientes pediátricos con queratocono por la diferencia de progresión entre ambos ojos. El seguimiento periódico permite diagnosticar la enfermedad en el ojo contralateral en pacientes con diagnóstico de queratocono en un ojo, indicar una corrección óptica individualizada, sobre todo en presencia de anisometropía, y monitorizar la evolución de la ambliopía refractiva asociada con frecuencia(AU)

Keratoconus is an asymmetric bilateral corneal ectasia, in early stages binocular involvement is not evident. Knowledge of the degree of asymmetry in pediatric keratoconus allows timely decisions in patient management. This research was carried out with the objective of describing the clinical characteristics and the treatment of three pediatric cases with different degree of interocular asymmetry of keratoconus, treated in the provincial consultation of pediatric corneal ectasia in Ciego de Avila. Patients showed difference between both eyes in clinical signs, refraction and topographic variables. Patient two was diagnosed with keratoconus in one eye, with no clinical or topographic evidence of the disease in the contralateral eye. All patients showed compound myopic astigmatism in both eyes and optical correction was indicated. Patient one was corrected with glasses, patient two with rigid gas permeable contact lenses and patient three with piggyback in one eye and rigid gas permeable contact lenses in the other eye. They also presented an associated amblyopia and were directed occlusive treatment and near vision tasks in patients two and three. It is common to find interocular asymmetry in pediatric patients with keratoconus due to the difference in progression between the two eyes. Periodic follow-up makes it possible to diagnose the disease in the contralateral eye in patients diagnosed with keratoconus in one eye, to indicate individualized optical correction, especially in the presence of anisometropia, and to monitor the evolution of frequently associated refractive amblyopia(AU)

Insulina colirio en afecciones de la superficie ocular / Insulin Eye Drops for Eye Surface Conditions

Cuando se produce una erosión corneal y fracasa la epitelización corneal surgen los defectos epiteliales corneales persistentes, cuyo tratamiento es un desafío para el oftalmólogo. Es muy frecuente el fracaso del tratamiento convencional por lo que se mantiene el interés en la búsqueda de otros factores de crecimiento para la cicatrización epitelial tales como los colirios de insulina. La insulina es un péptido estrechamente relacionado con el factor de crecimiento similar a la insulina 1. Su mecanismo de acción no es bien comprendido, sin embargo se acepta que es capaz de inducir migración y proliferación de las células epiteliales corneales, por lo que promueve y acelera la reepitelización de defectos epiteliales persistentes refractarios a tratamiento. La ausencia de una presentación comercial de colirio de insulina, hace necesario conocer su estabilidad físicoquímica y microbiológica así como la eficacia, efectividad y seguridad del colirio de insulina a diferentes concentraciones. De ahí la motivación para realizar una revisión de la literatura existente sobre el empleo del colirio de insulina en el tratamiento del defecto epitelial corneal persistente. Se realizó la búsqueda en bases de datos electrónicas como PubMed Central, EBSCO, Clinical Trials.gov, MEDLINE OVID, EMBASE OVID con el objeto de identificar artículos relacionados con el tema(AU)

When corneal erosion occurs and corneal epithelialization fails, persistent corneal epithelial defects arise, whose treatment is a challenge for the ophthalmologist. The failure of conventional treatment is very frequent; therefore, there is still interest in the search for other growth factors for epithelial healing, such as insulin eye drops. Insulin is a peptide closely related to insulin-like growth factor 1. Its mechanism of action is not well understood; however, it is accepted that it is capable of inducing migration and proliferation of corneal epithelial cells, thereby promoting and accelerating reepithelialization of persistent epithelial defects refractory to treatment. The absence of a commercial presentation for insulin eye drops makes it necessary to know its physicochemical and microbiological stability, as well as the efficacy, effectiveness and safety of insulin eye drops at different concentrations; hence the motivation to review the existing literature on the use of insulin eye drops in the treatment of persistent corneal epithelial defects. The search was carried out in electronic databases such as PubMed Central, EBSCO, Clinical Trials.gov, MEDLINE OVID, EMBASE OVID, with the aim of identifying relevant articles related to the topic(AU)

Effects of a laboratory-based aerobic exercise intervention on brain volume and cardiovascular health markers: protocol for a randomised clinical trial.

INTRODUCTION: Physical activity (PA) has beneficial effects on brain health and cardiovascular disease (CVD) risk. Yet, we know little about whether PA-induced changes to physiological mediators of CVD risk influence brain health and whether benefits to brain health may also explain PA-induced improvements to CVD risk. This study combines neurobiological and peripheral physiological methods in the context of a randomised clinical trial to better understand the links between exercise, brain health and CVD risk. METHODS AND ANALYSIS: In this 12-month trial, 130 healthy individuals between the ages of 26 and 58 will be randomly assigned to either: (1) moderate-intensity aerobic PA for 150 min/week or (2) a health information control group. Cardiovascular, neuroimaging and PA measurements will occur for both groups before and after the intervention. Primary outcomes include changes in (1) brain structural areas (ie, hippocampal volume); (2) systolic blood pressure (SBP) responses to functional MRI cognitive stressor tasks and (3) heart rate variability. The main secondary outcomes include changes in (1) brain activity, resting state connectivity, cortical thickness and cortical volume; (2) daily life SBP stress reactivity; (3) negative and positive affect; (4) baroreflex sensitivity; (5) pulse wave velocity; (6) endothelial function and (7) daily life positive and negative affect. Our results are expected to have both mechanistic and public health implications regarding brain-body interactions in the context of cardiovascular health. ETHICS AND DISSEMINATION: Ethical approval has been obtained from the University of Pittsburgh Institutional Review Board (IRB ID: 19020218). This study will comply with the NIH Data Sharing Policy and Policy on the Dissemination of NIH-Funded Clinical Trial Information and the Clinical Trials Registration and Results Information Submission rule. TRIAL REGISTRATION NUMBER: NCT03841669.

A Novel Imaging Protocol for Investigating Arabidopsis thaliana Siliques and Seeds Using X-rays.

Understanding silique and seed morphology is essential to developmental biology. Arabidopsis thaliana is one of the best-studied plant models for understanding the genetic determinants of seed count and size. However, the small size of its seeds, and their encasement in a pod known as silique, makes investigating their numbers and morphology both time consuming and tedious. Researchers often report bulk seed weights as an indicator of average seed size, but this overlooks individual seed details. Removal of the seeds and subsequent image analysis is possible, but automated counts are often impossible due to seed pigmentation and shadowing. Traditional ways of analyzing seed count and size, without their removal from the silique, involve lengthy histological processing (24-48 h) and the use of toxic organic solvents. We developed a method that is non-invasive, requires minimal sample processing, and obtains data in a short period of time (1-2 h). This method uses a custom X-ray imaging system to visualize Arabidopsis siliques at different stages of their growth. We show that this process can be successfully used to analyze the overall topology of Arabidopsis siliques and seed size and count. This new method can be easily adapted for other plant models. Key features • No requirement for organic solvents for imaging siliques. • Easy image capture and rapid turnaround time for obtaining data. • Protocol may be easily adapted for other plant models.

Use of Artificial Intelligence in the Identification and Diagnosis of Frailty Syndrome in Older Adults: Scoping Review.

BACKGROUND: Frailty syndrome (FS) is one of the most common noncommunicable diseases, which is associated with lower physical and mental capacities in older adults. FS diagnosis is mostly focused on biological variables; however, it is likely that this diagnosis could fail owing to the high biological variability in this syndrome. Therefore, artificial intelligence (AI) could be a potential strategy to identify and diagnose this complex and multifactorial geriatric syndrome. OBJECTIVE: The objective of this scoping review was to analyze the existing scientific evidence on the use of AI for the identification and diagnosis of FS in older adults, as well as to identify which model provides enhanced accuracy, sensitivity, specificity, and area under the curve (AUC). METHODS: A search was conducted using PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews) guidelines on various databases: PubMed, Web of Science, Scopus, and Google Scholar. The search strategy followed Population/Problem, Intervention, Comparison, and Outcome (PICO) criteria with the population being older adults; intervention being AI; comparison being compared or not to other diagnostic methods; and outcome being FS with reported sensitivity, specificity, accuracy, or AUC values. The results were synthesized through information extraction and are presented in tables. RESULTS: We identified 26 studies that met the inclusion criteria, 6 of which had a data set over 2000 and 3 with data sets below 100. Machine learning was the most widely used type of AI, employed in 18 studies. Moreover, of the 26 included studies, 9 used clinical data, with clinical histories being the most frequently used data type in this category. The remaining 17 studies used nonclinical data, most frequently involving activity monitoring using an inertial sensor in clinical and nonclinical contexts. Regarding the performance of each AI model, 10 studies achieved a value of precision, sensitivity, specificity, or AUC ≥90. CONCLUSIONS: The findings of this scoping review clarify the overall status of recent studies using AI to identify and diagnose FS. Moreover, the findings show that the combined use of AI using clinical data along with nonclinical information such as the kinematics of inertial sensors that monitor activities in a nonclinical context could be an appropriate tool for the identification and diagnosis of FS. Nevertheless, some possible limitations of the evidence included in the review could be small sample sizes, heterogeneity of study designs, and lack of standardization in the AI models and diagnostic criteria used across studies. Future research is needed to validate AI systems with diverse data sources for diagnosing FS. AI should be used as a decision support tool for identifying FS, with data quality and privacy addressed, and the tool should be regularly monitored for performance after being integrated in clinical practice.