Transplanted human photoreceptors transfer cytoplasmic material but not to the recipient mouse retina.

BACKGROUND: The discovery of material transfer between transplanted and host mouse photoreceptors has expanded the possibilities for utilizing transplanted photoreceptors as potential vehicles for delivering therapeutic cargo. However, previous research has not directly explored the capacity for human photoreceptors to engage in material transfer, as human photoreceptor transplantation has primarily been investigated in rodent models of late-stage retinal disease, which lack host photoreceptors. METHODS: In this study, we transplanted human stem-cell derived photoreceptors purified from human retinal organoids at different ontological ages (weeks 10, 14, or 20) into mouse models with intact photoreceptors and assessed transfer of human proteins and organelles to mouse photoreceptors. RESULTS: Unexpectedly, regardless of donor age or mouse recipient background, human photoreceptors did not transfer material in the mouse retina, though a rare subset of donor cells (< 5%) integrated into the mouse photoreceptor cell layer. To investigate the possibility that a species barrier impeded transfer, we used a flow cytometric assay to examine material transfer in vitro. Interestingly, dissociated human photoreceptors transferred fluorescent protein with each other in vitro, yet no transfer was detected in co-cultures of human and mouse photoreceptors, suggesting that material transfer is species specific. CONCLUSIONS: While xenograft models are not a tractable system to study material transfer of human photoreceptors, these findings demonstrate that human retinal organoid-derived photoreceptors are competent donors for material transfer and thus may be useful to treat retinal degenerative disease.

Small-Molecule-Directed Endogenous Regeneration of Visual Function in a Mammalian Retinal Degeneration Model.

Degenerative retinal diseases associated with photoreceptor loss are a leading cause of visual impairment worldwide, with limited treatment options. Phenotypic profiling coupled with medicinal chemistry were used to develop a small molecule with proliferative effects on retinal stem/progenitor cells, as assessed in vitro in a neurosphere assay and in vivo by measuring Msx1-positive ciliary body cell proliferation. The compound was identified as having kinase inhibitory activity and was subjected to cellular pathway analysis in non-retinal human primary cell systems. When tested in a disease-relevant murine model of adult retinal degeneration (MNU-induced retinal degeneration), we observed that four repeat intravitreal injections of the compound improved the thickness of the outer nuclear layer along with the regeneration of the visual function, as measured with ERG, visual acuity, and contrast sensitivity tests. This serves as a proof of concept for the use of a small molecule to promote endogenous regeneration in the eye.

Sweet potato flour (Ipomoea batatas) as natural antioxidant on carcass yield and meat physicochemical characteristics of Creole chickens of Mexico and Cobb 500.

Antioxidants are considered functional additives against oxidative stress since they avoid nutritional decline in the meat. The main objective of the present study is to evaluate the effect of sweet potato flour (SPF) as a natural antioxidant on carcass yield and physicochemical characteristics of Creole chickens of Mexico (CChM) and Cobb 500 broilers. In total, 210 chickens (105 CChM and 105 Cobb 500 chickens) were randomly assigned to three treatments: 0, 500, and 1000 mg of SPF kg-1 of feed. The Cobb 500 chickens showed higher carcass yield (hot and cold), breast, and breast fillet, whereas the CChM had higher thigh yield (P ≤ 0.05). The yield on the previously mentioned variables was not affected by the inclusion levels of SPF. The initial pH differed because of the effect of the chicken's genotype and the addition of SPF, which was higher on Cobb 500 chicken and on those that were not supplemented with SPF. The birds' skin that consumed SPF presented higher yellowness after 24 h (P ≤ 0.05). CChM manifested a higher dry matter and protein content and a lower content of ash and fat (P ≤ 0.05). In conclusion, Cobb 500 chickens present a higher carcass yield and its components, in addition to a less acid pH; however, CChM offer a higher nutritional contribution, whereas the 500 and 1000 mg addition of SPF increases the skin yellowness, which makes it an alterorganic as a pigment on broiler chicken production.

Progenitor division and cell autonomous neurosecretion are required for rod photoreceptor sublaminar positioning.

Migration is essential for the laminar stratification and connectivity of neurons in the central nervous system. In the retina, photoreceptors (PRs) migrate to positions according to birthdate, with early-born cells localizing to the basal-most side of the outer nuclear layer. It was proposed that apical progenitor mitoses physically drive these basal translocations non-cell autonomously, but direct evidence is lacking, and whether other mechanisms participate is unknown. Here, combining loss- or gain-of-function assays to manipulate cell cycle regulators (Sonic hedgehog, Cdkn1a/p21) with an in vivo lentiviral labelling strategy, we demonstrate that progenitor division is one of two forces driving basal translocation of rod soma. Indeed, replacing Shh activity rescues abnormal rod translocation in retinal explants. Unexpectedly, we show that rod differentiation also promotes rod soma translocation. While outer segment function or formation is dispensable, Crx and SNARE-dependent synaptic function are essential. Thus, both non-cell and cell autonomous mechanisms underpin PR soma sublaminar positioning in the mammalian retina.

Designing nutrition-sensitive agriculture (NSA) interventions with multi-criteria decision analysis (MCDA): a review.

Despite the efforts to end malnutrition through intensive agriculture of caloric crops, micronutrient deficiencies and other forms of malnutrition persist in vulnerable communities worldwide. Nutrition-sensitive agriculture (NSA) interventions are recognized as chances to address the causes of malnutrition. In this work, the different types of NSA interventions were explored, as well as the pathways through which they can improve nutrition (e.g., increasing biofortified crops and income generation via agricultural sales for a positive impact on access to nutritious foods, and simultaneously involving nutrition education to improve care practices and eventually nutritional status). Some NSA interventions focus on one pathway. Well-designed interventions, however, should follow multi-pathway approaches targeting the underlying causes of undernutrition within the selected population. The circumstances in which certain indicators should be used to measure the impact of an NSA intervention in each stage of the full pathway were also explained, as well as the need of enhancing the design of such interventions. Multi-criteria decision analysis (MCDA) has been employed to solve agriculture-related issues, but it has not been used to identify the optimal types of NSA interventions, metrics, and indicators based on the context of the community, priorities and objectives of the project managers and designers, etc.

Meningeal macrophages inhibit chemokine signaling in pre-tumor cells to suppress mouse medulloblastoma initiation.

The microenvironment profoundly influences tumor initiation across numerous tissues but remains understudied in brain tumors. In the cerebellum, canonical Wnt signaling controlled by Norrin/Frizzled4 (Fzd4) activation in meningeal endothelial cells is a potent inhibitor of preneoplasia and tumor progression in mouse models of Sonic hedgehog medulloblastoma (Shh-MB). Single-cell transcriptome profiling and phenotyping of the meninges indicate that Norrin/Frizzled4 sustains the activation of meningeal macrophages (mMΦs), characterized by Lyve1 and CXCL4 expression, during the critical preneoplastic period. Depleting mMΦs during this period enhances preneoplasia and tumorigenesis, phenocopying the effects of Norrin loss. The anti-tumorigenic function of mMΦs is derived from the expression of CXCL4, which counters CXCL12/CXCR4 signaling in pre-tumor cells, thereby inhibiting cell-cycle progression and promoting migration away from the pre-tumor niche. These findings identify a pivotal role for mMΦs as key mediators in chemokine-regulated anti-cancer crosstalk between the stroma and pre-tumor cells in the control of MB initiation.

Hydrogel assisted photoreceptor delivery inhibits material transfer.

Cell therapy holds tremendous promise for vision restoration; yet donor cell survival and integration continue to limit efficacy of these strategies. Transplanted photoreceptors, which mediate light sensitivity in the retina, transfer cytoplasmic components to host photoreceptors instead of integrating into the tissue. Donor cell material transfer could, therefore, function as a protein augmentation strategy to restore photoreceptor function. Biomaterials, such as hyaluronan-based hydrogels, can support donor cell survival but have not been evaluated for effects on material transfer. With increased survival, we hypothesized that we would achieve greater material transfer; however, the opposite occurred. Photoreceptors delivered to the subretinal space in mice in a hyaluronan and methylcellulose (HAMC) hydrogel showed reduced material transfer. We examined mitochondria transfer in vitro and cytosolic protein transfer in vivo and demonstrate that HAMC significantly reduced transfer in both contexts, which we ascribe to reduced cell-cell contact. Nanotube-like donor cell protrusions were significantly reduced in the hydrogel-transplanted photoreceptors compared to the saline control group, which suggests that HAMC limits the contact required to the host retina for transfer. Thus, HAMC can be used to manipulate the behaviour of transplanted donor cells in cell therapy strategies.

Risk Factors, Clinical Presentation, Therapeutic Trends, and Outcomes in Arterial Thrombosis Complicating Unvaccinated COVID-19 Patients: A Systematic Review.

Data on characteristics and outcomes of coronavirus (COVID)-19 patients complicated with arterial thrombosis (AT) are scarce. Therefore, we carried out a systematic review (PRISMA, PROSPERO statements; PubMed, Scopus, and Web of Science) to identify risk factors, clinical presentation, treatment, and outcomes. We included publications from December 2019 to October 2020. Groups: (a) ischemic stroke, (b) thrombotic storm, (c) peripheral vascular thrombosis, (d) myocardial infarction, and (e) left cardiac thrombus or in-transit thrombus (venous system thrombus floating or attaching to the right heart). We considered 131 studies. The most frequent cardiovascular risk factors were: hypertension, diabetes, and dyslipidemia. A high proportion presented with asymptomatic, mild, or moderate COVID-19 (n = 91, 41.4%). We identified a high percentage of isolated ischemic stroke and thrombotic storm. Groups with higher mortality rate: intracardiac thrombus (1/2, 50.0%), thrombotic storm (18/49, 36.7%), and ischemic stroke (48/131, 36.6%). A small number received thromboprophylaxis. Most patients received antithrombotic treatment. The most frequent bleeding complication was intracranial hemorrhage, primarily with isolated stroke. Overall mortality was 33.6% (74/220). Despite a wide range of COVID-19 severity, a high proportion had AT as a complication of non-severe disease. AT can affect different vascular territories; mortality is associated with stroke, intensive care unit stay, and severe COVID-19.

Objective Evaluation of Laparoscopic Experience Based on Muscle Electromyography and Accelerometry Performing Circular Pattern Cutting Tasks: A Pilot Study.

Purpose.The aim of this work is to present a new physical laparoscopy simulator with an electromyography (EMG)/accelerometry-based muscle activity recording system, EvalLap EMG-ACC, and perform objective evaluation of laparoscopic skills based on the quantification of muscle activity of participants with different levels of laparoscopic experience. Methods. EMG and ACC signals were obtained from 14 participants (6 experts, 8 medical students) performing circular pattern cutting tasks using a laparoscopic box trainer with the Trigno (Delsys Inc, Natick, MA) portable wireless system of 16 wireless sensors. Sensors were placed on the proximal and distal muscles of the upper extremities. Seven evaluation metrics were proposed and compared between skilled and novice surgeons. Results. The proximal and distal arm muscles (trapezius, deltoids, biceps, and forearms) were most active while executing laparoscopic tasks. Laparoscopic experience was associated with differences in EMG amplitude (Aavg), muscle activity (iEMG), hand acceleration (iACH), user movement (iAC), and muscle fatigue. For the cutting task, the deltoid, bicep, forearm EMG amplitude, and user movement significantly differed between experience groups. Conclusion. This pilot study demonstrates that different muscle groups are preferentially activated during laparoscopic tasks depending on the level of surgical experience. Expert surgeons showed less muscle activity compared with novices. EvalLap EMG-ACC represents a promising means to distinguish surgeons with basic cutting skills from those who have not yet developed these skills.

Objective psychomotor laparoscopic skills evaluation using a low-cost wearable device based on accelerometry: construct and concurrent validity study.

BACKGROUND: Motion analysis of surgical maneuvers provides useful quantitative information for the objective evaluation of the surgeons. However, surgical simulation laboratories for laparoscopic training do not usually integrate devices that help quantify the level of skills of the surgeons due to their limited resources and the high costs of new technologies. The purpose of this study is to present the construct and concurrent validity of a low-cost motion tracking system, based on a wireless triaxial accelerometer, employed to objectively evaluate psychomotor skills of surgeons during laparoscopic training. METHODS: An accelerometry system, a wireless three-axis accelerometer with appearance of wristwatch, was placed on the dominant hand of the surgeons to register the motion during the laparoscopy practice with the EndoViS simulator, which simultaneously recorded the motion of the laparoscopic needle driver. This study included the participation of 30 surgeons (6 experts, 14 intermediates and 10 novices) who performed the task of intracorporeal knot-tying suture. Using 11 motion analysis parameters (MAPs), the performance of each participant was assessed. Subsequently, the scores of the three groups of surgeons were statistically analyzed. In addition, a validity study was conducted comparing the metrics between the accelerometry-tracking system and the EndoViS hybrid simulator. RESULTS: Construct validity was achieved for 8 of the 11 metrics examined with the accelerometry system. Concurrent validity demonstrated that there is a strong correlation between the results of the accelerometry system and the EndoViS simulator in 9 of 11 parameters, showing reliability of the accelerometry system as an objective evaluation method. CONCLUSION: The accelerometry system was successfully validated. This method is potentially useful to complement the objective evaluation of surgeons during laparoscopic practice in training environments such as box-trainers and simulators.

Regulation of MAPK Signaling Pathways by the Large HERC Ubiquitin Ligases.

Protein ubiquitylation acts as a complex cell signaling mechanism since the formation of different mono- and polyubiquitin chains determines the substrate's fate in the cell. E3 ligases define the specificity of this reaction by catalyzing the attachment of ubiquitin to the substrate protein. Thus, they represent an important regulatory component of this process. Large HERC ubiquitin ligases belong to the HECT E3 protein family and comprise HERC1 and HERC2 proteins. The physiological relevance of the Large HERCs is illustrated by their involvement in different pathologies, with a notable implication in cancer and neurological diseases. Understanding how cell signaling is altered in these different pathologies is important for uncovering novel therapeutic targets. To this end, this review summarizes the recent advances in how the Large HERCs regulate the MAPK signaling pathways. In addition, we emphasize the potential therapeutic strategies that could be followed to ameliorate the alterations in MAPK signaling caused by Large HERC deficiencies, focusing on the use of specific inhibitors and proteolysis-targeting chimeras.

Growth performance, carcass, and meat quality traits of Tropical Milking criollo bulls fed with two energy-level diets.

The aim of this study was to determine the effect of two energy-level diets on the beef performance of Tropical Milking criollo bulls grouped by weight and age in three periods: initial, middle, and final. Evidence on intensive beef production from Tropical Milking criollo cattle breeds are unknown, and productive alternatives for dairy producers in the hot tropics are needed. Bulls were fed in each period with low (LE, 2.2 Mcal ME/kg DM) and high (HE, 2.9 Mcal ME/kg DM) energy diets and content phase feeding protein of 14.5, 12.0, and 10.3%, respectively. For each period, ten bulls were randomly assigned to diets. In all periods, growth performance for final body weight, average daily gain, feed intake, and feed conversion were all superior in HE (p ≤ 0.05). For the final period, slaughter body, hot and cold carcass weights, and dressing were also superior in HE (p ≤ 0.05); for meat quality traits, differences were observed in muscles Longissimus dorsi, Semimembranosus, Infraspinatus, and Serratus ventralis for pH and tenderness (p ≤ 0.05), although not for diets. For color L*, a*, and b* system, neither diet nor muscle had effect on L* (p > 0.05); however, the diet-muscle interaction was significant for a* (p ≤ 0.05), and only muscle effect was detected for b* (p ≤ 0.05). Tropical Milking bulls responded positively to a high-energy diet, despite not being a beef breed.

HERC1 deficiency causes osteopenia through transcriptional program dysregulation during bone remodeling.

Bone remodeling is a continuous process between bone-forming osteoblasts and bone-resorbing osteoclasts, with any imbalance resulting in metabolic bone disease, including osteopenia. The HERC1 gene encodes an E3 ubiquitin ligase that affects cellular processes by regulating the ubiquitination of target proteins, such as C-RAF. Of interest, an association exists between biallelic pathogenic sequence variants in the HERC1 gene and the neurodevelopmental disorder MDFPMR syndrome (macrocephaly, dysmorphic facies, and psychomotor retardation). Most pathogenic variants cause loss of HERC1 function, and the affected individuals present with features related to altered bone homeostasis. Herc1-knockout mice offer an excellent model in which to study the role of HERC1 in bone remodeling and to understand its role in disease. In this study, we show that HERC1 regulates osteoblastogenesis and osteoclastogenesis, proving that its depletion increases gene expression of osteoblastic makers during the osteogenic differentiation of mesenchymal stem cells. During this process, HERC1 deficiency increases the levels of C-RAF and of phosphorylated ERK and p38. The Herc1-knockout adult mice developed imbalanced bone homeostasis that presented as osteopenia in both sexes of the adult mice. By contrast, only young female knockout mice had osteopenia and increased number of osteoclasts, with the changes associated with reductions in testosterone and dihydrotestosterone levels. Finally, osteocytes isolated from knockout mice showed a higher expression of osteocytic genes and an increase in the Rankl/Opg ratio, indicating a relevant cell-autonomous role of HERC1 when regulating the transcriptional program of bone formation. Overall, these findings present HERC1 as a modulator of bone homeostasis and highlight potential therapeutic targets for individuals affected by pathological HERC1 variants.

Face, content, and construct validity of the virtual immersive operating room simulator for training laparoscopic procedures.

BACKGROUND: The aim of this work is to present the face, content, and construct validation of the virtual immersive operating room simulator (VIORS) for procedural training of surgeons' laparoscopic psychomotor skills and evaluate the immersive training experience. METHODS: The VIORS simulator consists of an HMD Oculus Rift 2016 with a visor on a 1080 × 1200 pixel OLED screen, two positioning sensors with two adapted controls to simulate laparoscopic instruments, and an acrylic base to simulate the conventional laparoscopic setup. The immersion consists of a 360° virtual operating room environment, based on the EndoSuite at Hospital Infantil de Mexico Federico Gomez, which reproduces a configuration of equipment, instruments, and common distractions in the operating room during a laparoscopic cholecystectomy procedure. Forty-five surgeons, residents, and medicine students participated in this study: 27 novices, 13 intermediates, and 5 experts. They completed a questionnaire on the realism and operating room immersion, as well as their capabilities for laparoscopic procedural training, scored in the 5-point Likert scale. The data of instrument movement were recorded and analyzed using 13 movement analysis parameters (MAPs). The experience during training with VIORS was evaluated through NASA-TLX. RESULTS: The participants were enthusiastic about the immersion and sensation levels of the VIORS simulator, with positive scores on the realism and its capabilities for procedural training using VIORS. The results proved that the VIORS simulator was able to differentiate between surgeons with different skill levels. Statistically significant differences were found in nine MAPs, demonstrating their construct validity for the objective assessment of the procedural laparoscopic performance. At cognitive level, the inversion experience proves a moderate mental workload when the laparoscopic procedure is carried out. CONCLUSION: The VIORS simulator has been successfully presented and validated. The VIORS simulator is a useful and effective device for the training of procedural laparoscopic psychomotor skills.

Path Forward to Optimise Post-approval Change Management and Facilitate Continuous Supply of Medicines and Vaccines of High Quality Worldwide : Joint Position from EFPIA, IFPMA and Vaccines Europe.

Post-approval changes (PACs) to the registered information of authorised medicinal products are introduced routinely worldwide to enhance the robustness and efficiency of the manufacturing process, ensure timely supply in case of increased demand, improve quality control techniques, respond to changes in regulatory requirements and upgrade to state-of-the-art facilities. These are critical to prevent supply disruption and continuously improve existing medicines and vaccines. Due to the complexity of current PAC systems across markets, a change can take 3 to 5 years to approval globally (Hoath et al in BioProcess Int, 2016) thus hindering innovation and increasing the risk of shortages. The key messages are as follows: 1. Industry believes that global regulatory convergence of post-approval changes to Marketing Authorisations (MAs) using science- and risk-based approaches will enable a more efficient management of quality and supply improvements and will facilitate patients' access to innovative medicines and vaccines of the highest quality. 2. National Regulatory Authorities (NRAs) should establish national or regional guidelines in line with international standards (regarding a risk-based classification of changes and standardisation of requirements) (Guidelines on procedures and data requirements for changes to approved biotherapeutic products, in WHO Technical Report Series, 2018, Guidelines on procedures and data requirements for changes to approved vaccines, in WHO Technical Report Series, 2015), have clear procedural guidance including timelines and implement reliance pathways to accelerate the approval of changes. This paper briefly outlines the challenges for PACs and provides solutions for a more flexible and aligned global system.

Zapatilla distal modificada por pérdida prematura de guía eruptiva. Presentación de un caso clínico / Chinelo distal modificado devido à perda prematura da orientação eruptiva. Apresentação de um caso clínico / Distal shoe due to premature loss of eruptive guidance. Presentation of a clinical case

La oclusión se establece como resultado de la interacción de factores genéticos y ambientales. Es de suma importancia la dentición primaria para el mantenimiento de la oclusión, estética, fonética y bienestar psicoemocional del niño. La migración del primer molar permanente ante la ausencia del segundo molar primario provoca la pérdida localizada del espacio. El mantenedor de espacio llamado zapatilla distal, resulta ser beneficioso para restablecer la erupción del primer molar permanente no erupcionado. Objetivo. Restablecer la guía eruptiva del primer molar permanente posterior a la pérdida prematura del segundo y primer molar primarios, así como la función y el estímulo masticatorio. Caso Clínico. Masculino de 5,4 años con pérdida prematura de primer y segundo molar primario inferior derecho debido al antecedente de proceso infeccioso abordado intrahospitalariamente 6 meses atrás. Previa rehabilitación bucal del paciente, se realizó aparatología fija tipo zapatilla distal con anclaje en corona de acero cromo modificada con molares de acrílico en zona edéntula, se realizó incisión en mucosa para el contacto de la zapatilla con la cara mesial del primer molar permanente no erupcionado. Restableciendo la guía eruptiva del primer molar permanente, función masticatoria y la dimensión vertical dentoalveolar del sector posterior. Conclusiones. Los diseños convencionales de los mantenedores de espacio no son una regla, se pueden modificar satisfaciendo las necesidades de atención del paciente, estableciendo un vínculo entre los padres y el paciente para un control radiográfico y seguimiento preventivo de la erupción y control de salud bucal.

A oclusão é estabelecida como resultado da interação de fatores genéticos e ambientais. A dentição decidua é de suma importância para a manutenção da oclusão, estética, fonética e bem-estar psicoemocional da criança. A migração do primeiro molar permanente antes da ausência do segundo molar deciduo provoca a perda localizada do espaço. O mantenedor de espaço chamado zapatilla distal, resulta ser benéfico para restabelecer a erupção do primer molar permanente não erupcionado. Objetivo. Restabelecer a guia eruptiva do primer molar permanente devido à perda prematura do segundo e do primer molar deciduo, assim como a função e o estímulo mastigatório. Caso Clínico. Paciente masculino de 5,4 anos com perda prematura de primer e segundo molar deciduo inferior devido ao antecedente de processo infeccioso intra-hospitalar 6 meses atrás. Prévia reabilitação bucal do paciente, se realizou aparatología fija tipo zapatilla distal com anclaje em corona de aço cromo modificado com molares de acrílico na zona edêntula, se realizou incisão na mucosa para el contato da zapatilla com a cara mesial do primer molar permanente não erupcionado. Restabelecendo a guia de erupção do primer molar permanente, função mastigatória e a dimensão vertical dentoalveolar do setor posterior. Conclusão. Projetos convencionais de mantenedores de espaço não são regra, eles podem ser modificados para atender às necessidades de cuidados do paciente, estabelecendo um vínculo entre pais e paciente para monitoramento radiográfico e monitoramento preventivo de erupção e monitoramento da saúde bucal

The occlusion is established because of the interaction of genetic and environmental factors. Primary dentition is of paramount importance for the maintenance of occlusion, aesthetics, phonetics, and psycho-emotional well-being of the child. The migration of the first permanent molar in the absence of the second primary molar causes a localized loss of space. The space maintainer called the distal shoe, turns out to be beneficial for restoring the eruption of the non-erupted permanent first molar. Objective. Restore the eruptive guide of the first permanent molar due to the premature loss of the primary molars, as well as the function and the masticatory stimulus. Clinical case. A 5.4-year-old male with premature loss of a right primary lower molar by intra-hospital approach infectious process 6 months ago. Prior to the oral rehabilitation of the patient, a fixed apparatus of the distal shoe type was performed with an anchor in a chrome steel crown modified with acrylic molars in the edentulous area, an incision was made in the mucosa for the contact of the shoe with the mesial face of the non-erupted permanent first molar. Restoring the eruptive guide of the permanent first molar, masticatory function and the vertical dentoalveolar dimension of the posterior sector. Conclusions. The conventional designs of space maintainers are not a rule, they can be modified to satisfy the patient's attention needs, establishing a bond between parents and the patient for radiographic control and preventive follow-up of eruption and oral health monitoring

HERC2 deficiency activates C-RAF/MKK3/p38 signalling pathway altering the cellular response to oxidative stress.

HERC2 gene encodes an E3 ubiquitin ligase involved in several cellular processes by regulating the ubiquitylation of different protein substrates. Biallelic pathogenic sequence variants in the HERC2 gene are associated with HERC2 Angelman-like syndrome. In pathogenic HERC2 variants, complete absence or marked reduction in HERC2 protein levels are observed. The most common pathological variant, c.1781C > T (p.Pro594Leu), encodes an unstable HERC2 protein. A better understanding of how pathologic HERC2 variants affect intracellular signalling may aid definition of potential new therapies for these disorders. For this purpose, we studied patient-derived cells with the HERC2 Pro594Leu variant. We observed alteration of mitogen-activated protein kinase signalling pathways, reflected by increased levels of C-RAF protein and p38 phosphorylation. HERC2 knockdown experiments reproduced the same effects in other human and mouse cells. Moreover, we demonstrated that HERC2 and RAF proteins form molecular complexes, pull-down and proteomic experiments showed that HERC2 regulates C-RAF ubiquitylation and we found out that the p38 activation due to HERC2 depletion occurs in a RAF/MKK3-dependent manner. The displayed cellular response was that patient-derived and other human cells with HERC2 deficiency showed higher resistance to oxidative stress with an increase in the master regulator of the antioxidant response NRF2 and its target genes. This resistance was independent of p53 and abolished by RAF or p38 inhibitors. Altogether, these findings identify the activation of C-RAF/MKK3/p38 signalling pathway in HERC2 Angelman-like syndrome and highlight the inhibition of RAF activity as a potential therapeutic option for individuals affected with these rare diseases.

Effect of varying dietary crude protein and metabolizable energy levels on leg abnormalities and performance in broiler chickens

Background: Nutrient restriction is a common strategy to prevent metabolic disorders in fast growing broiler chickens, but limited information is available regarding the impact of diets with low protein and energy on leg abnormalities Objective: Two experiments were conducted to evaluate the effect of varying crude protein (CP) and metabolizable energy (ME) levels on gastrocnemius tendon (GTeBS) and tibia breaking strength (TiBS), gait score (GS) and valgus/varus angulation (VAng). Methods: In Experiment 1, eight-d-old Ross 308 broilers (n=90) were randomly assigned into three treatments: 13/2,900 (13% CP and 2,900 kcal ME kg-1 of diet), 17/3,000 (17% CP and 3,000 kcal ME kg-1 of diet), and 21/3,025 or control (21% CP and 3,025 kcal ME kg-1 of diet). In Experiment 2, six-d-old Ross 308 chickens (n=192) were randomly distributed into two treatments: 16/3,000; 16% CP and 3,000 kcal ME kg-1 of diet and 21/3,000 or control; 21% CP and 3,000 kcal ME kg-1 of diet. In both experiments data were analyzed as one-way ANOVA. Results: In Experiment 1, broilers in the 17/3,000 and control treatments had similar (p>0.05) GTeBS (202 and 224 N, respectively), and TiBS (338 and 332 N, respectively). Birds in the 13/2,900 treatment showed higher GS (lower walking ability; 80% of birds with score >3), and greater VAng (53% of birds with score >2) than broilers in the control treatment (0% of birds with GS score >3 and 0% of birds with VAng score >2). Experiment 2, broilers in the 16/3,000 showed similar VAng, GS and TiBS than birds in the control treatment. However, birds in the 16/3,000 treatment showed lower (-26%) GTeBS than birds in the control treatment (p<0.05). Conclusions: Feeding broilers with 16% CP and 3,000 kcal ME kg-1 of diet did not affect tibia breaking strength, gait score and valgus/varus angulation. A diet containing 16% CP or less and 2,900-3,000 kcal ME kg-1 reduces tendon and tibia breaking strength.

Antecedentes: La restricción de nutrientes es una estrategia común para prevenir trastornos metabólicos en pollos de engorde, sin embargo, se dispone de información limitada sobre el impacto de las dietas bajas en proteína y energía en las anomalías de piernas. Objetivo: Se llevaron a cabo dos experimentos para evaluar el efecto de niveles variables de proteína cruda (PC) y energía metabolizable (EM) en la fuerza de rotura del tendón del gastrocnemio (GTeBS) y tibia (TiBS), la evaluación de marcha (GS) y la angulación en valgus/varus (VAng). Métodos: En el Experimento 1, pollos de engorde Ross 308 de ocho días de edad (n = 90) se asignaron al azar en tres tratamientos: 13/2.900; 13% PC y 2.900 kcal ME kg-1 de dieta, 17/3.000; 17% PC y 3.000 kcal ME kg-1 de dieta y 21/3.025 o control; 21% PC y 3.025 kcal ME kg-1 de dieta. En el Experimento 2, pollos Ross 308 de seis días de edad (n = 192) se distribuyeron aleatoriamente en dos tratamientos: 16/3.000; 16% PC y 3.000 kcal ME kg-1 de dieta y 21/3.000 o control; 21% PC y 3.000 kcal ME kg-1 de dieta. En ambos experimentos los datos se analizaron como ANOVA de una vía. Resultados: En el Experimento 1, los pollos de los tratamientos 17/3.000 y control tuvieron GTeBS (202 y 224 N, respectivamente) y TiBS (338 y 332 N, respectivamente) similares (p>0,05). Las aves del tratamiento 13/2.900 mostraron mayor GS (menor capacidad para caminar; 80% de las aves con puntaje >3) y mayor VAng (53% de las aves con puntaje >2) que los pollos del tratamiento control (0% de aves con un puntaje GS >3 y 0% de aves con un puntaje VAng >2). En el Experimento 2, los pollos del 16/3.000 mostraron VAng, GS y TiBS similares a los de las aves del tratamiento control. Sin embargo, las aves del tratamiento 16/3.000 mostraron menor (-26%) GTeBS que las aves control (p<0,05). Conclusiones: La alimentación de pollos de engorde con 16% PC y 3.000 kcal EM kg-1 de dieta no afecta la resistencia a la ruptura de la tibia, la marcha y la angulación valgus/varus. La dieta formulada con 16% de PC o menos y 2.900-3.000 kcal de EM kg-1 reduce la resistencia a la ruptura del tendón y de la tibia.

Antecedentes: A restrição de nutrientes é uma estratégia comum para prevenir distúrbios metabólicos em frangos de corte, no entanto, há informações limitadas disponíveis sobre o impacto de dietas de baixa proteína e baixa energia nas anormalidades de pernas destas aves. Objetivo: Dois experimentos foram realizados para avaliar o efeito de diferentes níveis de proteína bruta (PB) e energia metabolizável (EM) em força de ruptura do tendão do músculo gastrocnêmio (GTeBS), forca de ruptura da tíbia (TiBS), gait score (GS) e angulação em valgus/varus (VAng). Métodos: Experimento 1, foram utilizados 308 frangos de corte machos da linhagem Ross com oito dias de idade (n = 90) distribuidos aleatoriamente em três tratamentos: dieta controle com 21% PB e 3.025 kcal EM kg-1 13/2.900; dieta con 13% PB e 2.900 kcal EM kg-1, y 17% PB e 3.000 kcal EM kg-1 . Experimento 2, foram utilizados 308 frangos de corte da linhagem Ross com seis dias de idade (n = 192) distribuídos aleatoriamente em dois tratamentos: dieta controle con 21% PB e 3.000 kcal EM kg-1 e dieta com 16% PB e 3.000 kcal EM kg-1 . Em ambos os experimentos, os dados foram submetidos a ANOVA. Resultados: Experimento 1, os frangos de corte submetidos ao dos tratamentos 17/3.000 e controle nao apresentaram diferencas significativas (p>0,05) para as variáveis GteBS (202 y 224 N, respectivamente) e TiBS (338 y 332 N, respectivamente). Os frangos submetidos ao 13/2.900 apresentaram maior frequência de GS (menor capacidade de locomocao; 80% das aves com pontuação>3) e maior VAng (53% das aves com pontuação>2) do que os frangos do tratamento controle (0% das aves com pontuação GS>3 e 0% das aves com pontuação VAng>2). Experimento 2, os frangos do 16/3.000 apresentaram VAng, GS e TiBS semelhantes aos das aves do tratamento controle. Entretanto, as aves do tratamento 16/3.000 apresentaram GTeBS mais baixo (-26%) em comparacao as aves do tratamento de controle (p<0,05). Conclusões: A dieta com 16% PB e 3.000 kcal EM kg-1 não afetou a forca de ruptura da tíbia, gaite score marcha e angulação em valgus/varus de frangos de corte. Uma dieta com níveis de 16% de PB ou menos e 2.900-3.000 kcal EM kg-1 reduziu a resistência à ruptura do tendão e a resistência de tibia de frangos de corte.

Diesel effects on some population attributes of Orthonychiurus folsomi Schäffer 1900 (Collembola: Onychiuridae) under laboratory conditions.

Soil contamination by hydrocarbons and its effects on population health and welfare is a growing concern, especially in urban environments with industrial activity. Indicator species complement the information obtained from the measurement of environment quality by using physicochemical variables. The objective of this study was to evaluate the response of a springtail species that could be potentially used as a bioindicator of hydrocarbon contaminated sites. We studied the effects of seven diesel concentrations on survival and other population parameters of Orthonychiurus folsomi under laboratory conditions; we evaluated the springtails response on survival, fecundity, population size, hatching and development time. Survival and fertility were analyzed by using generalized linear models (GLM). An ANOVA test was used to analyze the final population size and a Kruskal-Wallis test for oviposition, hatching, and development times. Results showed that diesel has negative effects on some population parameters. The most evident effects were recorded above 500 mg/kg concentrations. A significant decrease in survival, fertility, and population size was recorded in function of diesel concentration increase. The estimated LC50 for survival was 955 mg/kg. Oviposition and hatching time increased significantly when diesel concentrations were above 500 mg/kg, while development time increased slightly at low diesel concentrations but decreased at concentrations above 500 mg/kg. Population final size decreased as soil diesel concentrations increased. Due to the sensitive and rapid response of O. folsomi, it could be useful to detect diesel-contaminated soil, mainly in urban areas.

Directed Evolution Enables Simultaneous Controlled Release of Multiple Therapeutic Proteins from Biopolymer-Based Hydrogels.

With the advent of increasingly complex combination strategies of biologics, independent control over their delivery is the key to their efficacy; however, current approaches are hindered by the limited independent tunability of their release rates. To overcome these limitations, directed evolution is used to engineer highly specific, low affinity affibody binding partners to multiple therapeutic proteins to independently control protein release rates. As a proof-of-concept, specific affibody binding partners for two proteins with broad therapeutic utility: insulin-like growth factor-1 (IGF-1) and pigment epithelium-derived factor (PEDF) are identified. Protein-affibody binding interactions specific to these target proteins with equilibrium dissociation constants (KD ) between 10-7 and 10-8 m are discovered. The affibodies are covalently bound to the backbone of crosslinked hydrogels using click chemistry, enabling sustained, independent, and simultaneous release of bioactive IGF-1 and PEDF over 7 days. The system is tested with C57BL/6J mice in vivo, and the affibody-controlled release of IGF-1 results in sustained activity when compared to bolus IGF-1 delivery. This work demonstrates a new, broadly applicable approach to tune the release of therapeutic proteins simultaneously and independently and thus the way for precise control over the delivery of multicomponent therapies is paved.