Neuronal DNA repair reveals strategies to influence CRISPR editing outcomes.

Genome editing is poised to revolutionize treatment of genetic diseases, but poor understanding and control of DNA repair outcomes hinders its therapeutic potential. DNA repair is especially understudied in nondividing cells like neurons, which must withstand decades of DNA damage without replicating. This lack of knowledge limits the efficiency and precision of genome editing in clinically relevant cells. To address this, we used induced pluripotent stem cells (iPSCs) and iPSC-derived neurons to examine how postmitotic human neurons repair Cas9-induced DNA damage. We discovered that neurons can take weeks to fully resolve this damage, compared to just days in isogenic iPSCs. Furthermore, Cas9-treated neurons upregulated unexpected DNA repair genes, including factors canonically associated with replication. Manipulating this response with chemical or genetic perturbations allowed us to direct neuronal repair toward desired editing outcomes. By studying DNA repair in postmitotic human cells, we uncovered unforeseen challenges and opportunities for precise therapeutic editing.

Translation and linguistic validation of 24 PROMIS item banks into French.

PURPOSE: The Patient-Reported Outcome Measurement Information System (PROMIS®) was developed to provide reliable, valid, and normed item banks to measure health. The item banks provide standardized scores on a common metric allowing for individualized, brief assessment (computerized adaptive tests), short forms (e.g. heart failure specific), or profile assessments (e.g. PROMIS-29). The objective of this study was to translate and linguistically validate 24 PROMIS adult item banks into French and highlight cultural nuances arising during the translation process. METHODS: We used the FACIT translation methodology. Forward translation into French by two native French-speaking translators was followed by reconciliation by a third native French-speaking translator. A native English-speaking translator fluent in French then completed a back translation of the reconciled version from French into English. Three independent reviews by bilingual translators were completed to assess the clarity and consistency of terminology and equivalency across the English source and French translations. Reconciled versions were evaluated in cognitive interviews for conceptual and linguistic equivalence. RESULTS: Twenty-four adult item banks were translated: 12 mental health, 10 physical health, and two social health. Interview data revealed that 577 items of the 590 items translated required no revisions. Conceptual and linguistic differences were evident for 11 items that required iterations to improve conceptual equivalence and two items were revised to accurately reflect the English source. CONCLUSION: French translations of 24 item banks were created for routine clinical use and research. Initial translation supported conceptual equivalence and comprehensibility. Next steps will include validation of the item banks.

Hollow viscus perforation in blunt abdominal trauma: A 14-year experience from a trauma center.

BACKGROUND: Isolated perforations of hollow viscus (HV) represent less than 1% of injuries in blunt abdominal trauma (BAT). When they do present, they are generally due to high-impact mechanisms in the segments of the intestine that are fixed. The aim of this study is to determine the incidence of major HV injuries in BAT at the "Dr. Domingo Luciani" General Hospital (HDL), and address the literature gap regarding updated HV perforations following BAT, especially in low-income settings. METHODS: A retrospective review was conducted on the medical records of patients admitted to our trauma center with a diagnosis of complicated BAT with HV perforation over 14 years. RESULTS AND DISCUSSION: Seven hundred sixty-one patients were admitted under the diagnosis of BAT. Of them, 36.79% underwent emergency surgical resolution, and 6.04% had HV perforation as an operative finding. Almost half (44.44%) of these cases presented as a single isolated injury, while the remaining were associated with other intra-abdominal organ injuries. The most common lesions were Grade II-III jejunum and Grade I transverse colon, affecting an equal proportion of patients at 13.33%. In recent years, an increased incidence of HV injuries secondary to BAT has been observed. Despite this, in many cases, the diagnosis is delayed, so even in the presence of negative diagnostic studies, the surgical approach based on the trauma mechanism, hemodynamic status, and systematic reevaluation of the polytraumatized patient should prevail.

In vivo human T cell engineering with enveloped delivery vehicles.

Viruses and virally derived particles have the intrinsic capacity to deliver molecules to cells, but the difficulty of readily altering cell-type selectivity has hindered their use for therapeutic delivery. Here, we show that cell surface marker recognition by antibody fragments displayed on membrane-derived particles encapsulating CRISPR-Cas9 protein and guide RNA can deliver genome editing tools to specific cells. Compared to conventional vectors like adeno-associated virus that rely on evolved capsid tropisms to deliver virally encoded cargo, these Cas9-packaging enveloped delivery vehicles (Cas9-EDVs) leverage predictable antibody-antigen interactions to transiently deliver genome editing machinery selectively to cells of interest. Antibody-targeted Cas9-EDVs preferentially confer genome editing in cognate target cells over bystander cells in mixed populations, both ex vivo and in vivo. By using multiplexed targeting molecules to direct delivery to human T cells, Cas9-EDVs enable the generation of genome-edited chimeric antigen receptor T cells in humanized mice, establishing a programmable delivery modality with the potential for widespread therapeutic utility.

Dynamic pathogen detection and social feedback shape collective hygiene in ants.

Cooperative disease defense emerges as group-level collective behavior, yet how group members make the underlying individual decisions is poorly understood. Using garden ants and fungal pathogens as an experimental model, we derive the rules governing individual ant grooming choices and show how they produce colony-level hygiene. Time-resolved behavioral analysis, pathogen quantification, and probabilistic modeling reveal that ants increase grooming and preferentially target highly-infectious individuals when perceiving high pathogen load, but transiently suppress grooming after having been groomed by nestmates. Ants thus react to both, the infectivity of others and the social feedback they receive on their own contagiousness. While inferred solely from momentary ant decisions, these behavioral rules quantitatively predict hour-long experimental dynamics, and synergistically combine into efficient colony-wide pathogen removal. Our analyses show that noisy individual decisions based on only local, incomplete, yet dynamically-updated information on pathogen threat and social feedback can lead to potent collective disease defense.

A Technical Note of Improvement of the Elnady Technique for Tissue Preservation in Veterinary Anatomy.

Teaching veterinary anatomy has been subjected to changes and restrictions that have promoted the development of new techniques for preserving organs and cadavers. The Elnady technique is a recent method for the conservation of tissues. Specimens produced with this technique are realistic, durable, soft, and flexible, but an undesirable feature is the discoloration of tissues. In the present study, we describe modifications of the Elnady technique for organ and tissue preservation. Specimens were prepared on the theoretical basis of the Elnady technique, but at low temperatures and with longer durations for the fixation, dehydration, glycerin impregnation and curing processes. Furthermore, the tissues were pigmented with a red vegetable pigment before dehydration or in the glycerin impregnation process. The results show high-quality specimens with minimal shrinkage and natural color aspects. The modified Elnady technique is adequate for producing specimens of better contrast for education purposes.

Yellowing, Weathering and Degradation of Marine Pellets and Their Influence on the Adsorption of Chemical Pollutants.

Marine microplastics (MPs) are exposed to environmental factors, which produce aging, weathering, surface cracking, yellowing, fragmentation and degradation, thereby changing the structure and behavior of the plastic. This degradation also has an influence on the adsorption of persistent organic pollutants over the microplastic surface, leading to increased concentration with aging. The degradation state affects the microplastic color over time; this is called yellowing, which can be quantified using the Yellowness Index (YI). Weathering and surface cracking is also related with the microplastic yellowing, which can be identified by Fourier transform infrared spectroscopy (FTIR). In this study, the degradation state of marine microplastic polyethylene pellets with different aging stages is evaluated and quantified with YI determination and the analysis of FTIR spectrums. A color palette, which relates to the microplastic color and YI, was developed to obtain a visual percentage of this index. The relation with the adsorption rate of persistent organic pollutant over the microplastic surface was also determined.

Early queen infection shapes developmental dynamics and induces long-term disease protection in incipient ant colonies.

Infections early in life can have enduring effects on an organism's development and immunity. In this study, we show that this equally applies to developing 'superorganisms'--incipient social insect colonies. When we exposed newly mated Lasius niger ant queens to a low pathogen dose, their colonies grew more slowly than controls before winter, but reached similar sizes afterwards. Independent of exposure, queen hibernation survival improved when the ratio of pupae to workers was small. Queens that reared fewer pupae before worker emergence exhibited lower pathogen levels, indicating that high brood rearing efforts interfere with the ability of the queen's immune system to suppress pathogen proliferation. Early-life queen pathogen exposure also improved the immunocompetence of her worker offspring, as demonstrated by challenging the workers to the same pathogen a year later. Transgenerational transfer of the queen's pathogen experience to her workforce can hence durably reduce the disease susceptibility of the whole superorganism.

CCG•CGG interruptions in high-penetrance SCA8 families increase RAN translation and protein toxicity.

Spinocerebellar ataxia type 8 (SCA8), a dominantly inherited neurodegenerative disorder caused by a CTG•CAG expansion, is unusual because most individuals that carry the mutation do not develop ataxia. To understand the variable penetrance of SCA8, we studied the molecular differences between highly penetrant families and more common sporadic cases (82%) using a large cohort of SCA8 families (n = 77). We show that repeat expansion mutations from individuals with multiple affected family members have CCG•CGG interruptions at a higher frequency than sporadic SCA8 cases and that the number of CCG•CGG interruptions correlates with age at onset. At the molecular level, CCG•CGG interruptions increase RNA hairpin stability, and in cell culture experiments, increase p-eIF2α and polyAla and polySer RAN protein levels. Additionally, CCG•CGG interruptions, which encode arginine interruptions in the polyGln frame, increase toxicity of the resulting proteins. In summary, SCA8 CCG•CGG interruptions increase polyAla and polySer RAN protein levels, polyGln protein toxicity, and disease penetrance and provide novel insight into the molecular differences between SCA8 families with high vs. low disease penetrance.

Morphometrical Study of the Lumbar Segment of the Internal Vertebral Venous Plexus in Dogs: A Contrast CT-Based Study.

The internal vertebral venous plexus (IVVP) is a thin-walled, valveless venous network that is located inside the vertebral canal, communicating with the cerebral venous sinuses. The objective of this study was to perform a morphometric analysis of the IVVP, dural sac, epidural space and vertebral canal between the L1 and L7 vertebrae with contrast-enhanced computed tomography (CT). Six clinically healthy adult dogs weighing between 12 kg to 28 kg were used in the study. The CT venographic protocol consisted of a manual injection of 880 mgI/kg of contrast agent (587 mgI/kg in a bolus and 293 mgI/mL by continuous infusion). In all CT images, the dimensions of the IVVP, dural sac, and vertebral canal were collected. Dorsal reconstruction CT images showed a continuous rhomboidal morphological pattern for the IVVP. The dural sac was observed as a rounded isodense structure throughout the vertebral canal. The average area of the IVVP ranged from 0.61 to 0.74 mm2 between L1 and L7 vertebrae (6.3-8.9% of the vertebral canal), and the area of the dural sac was between 1.22 and 7.42 mm2 (13.8-72.2% of the vertebral canal). The area of the epidural space between L1 and L7 ranged from 2.85 to 7.78 mm2 (27.8-86.2% of the vertebral canal). This CT venography protocol is a safe method that allows adequate visualization and morphometric evaluation of the IVVP and adjacent structures.

Distribution and transport of microplastics in the upper 1150 m of the water column at the Eastern North Atlantic Subtropical Gyre, Canary Islands, Spain.

Nowadays it is widely known that pollution by microplastics (MP) at the open ocean covers immense areas. Buoyant plastics tend to accumulate in areas of convergence at the sea surface such as subtropical gyres, while non-buoyant plastics accumulate at the seafloor. However, previous studies have revealed that the total amount of plastic in the different oceans is not well correlated with the concentrations measured at the sea surface and the sea floor, evidencing a significant amount of missing plastic in the oceans. This deviation could be related to an underestimation of the role played by small fragments of plastic and fibers in the oceans. Furthermore, microplastic fragments with a density lower than the density of seawater have been gathered hundreds of meters below the sea surface in the Pacific Ocean due to their size and shape. The main objective of this study is to carry out, for the first time, an equivalent analysis along the water column for the Atlantic Ocean. In that sense, a total number of 51 samples were collected during four different oceanographic cruises between February and December 2019, from the sea surface down to 1150 m depth at the open ocean waters of the Canary Islands region (Spain). For each sample, 72 l of seawater were filtered on board with a mesh size of 100 µm, where the presence of microplastics has been clearly observed. Our results reveal the presence of microplastics at least up to 1150 m depth, at the Northeastern Atlantic Subtropical Gyre with noticeable seasonal differences. The spatial distribution of these small fragments and fibers at the water column is mainly related to the oceanic dynamics and mesoscale convective flows, overcoming the MP motion induced by their own buoyancy. Moreover, these microplastics have being transported by the ocean dynamics as passive drifters.

Factores diferenciales entre las etapas evolutivas de la isquemia mesentérica aguda / Differential factors between the acute mesenteric ischemia evolutive stages

RESUMEN Fundamento: la isquemia mesentérica aguda tiene una elevada letalidad debido a la rápida evolución de su etapa temprana a la tardía, la larvada sintomatología en su fase inicial y por la complejidad del tratamiento. Objetivo: determinar las diferencias clínicas, de laboratorio, imagenológicas y evolutivas entre las etapas de la isquemia mesentérica aguda. Métodos: se realizó un estudio observacional, analítico transversal que incluyó un universo de 32 pacientes con isquemia mesentérica aguda, desde diciembre de 2015 a marzo de 2018, en el Hospital General Universitario Vladimir Ilich Lenin. Resultados: la reacción peritoneal se asoció de forma muy significativa a la etapa avanzada. La mayor parte de los pacientes con isquemia intestinal no presentaron niveles hidroaéreos, al contrario de los diagnosticados con necrosis. Todos los afectados con fibrilación auricular se encontraron en la etapa tardía. El valor medio del pH fue mayor en la etapa inicial, mientras que el valor del lactato en la fase tardía duplicó el valor medio de los casos isquémicos. La diferencia de medias de estos valores entre ambas fases de la enfermedad fue estadísticamente muy significativa. Fallecieron todos los casos con necrosis intestinal y más de dos tercios de los afectados con isquemia intestinal. Conclusiones: el retardo en el diagnóstico se asoció a una elevada mortalidad. La sospecha clínica temprana de abdomen agudo, la valoración de parámetros hemodinámicos y la identificación de factores de riesgo como la fibrilación auricular, favorecen el tratamiento oportuno en etapa isquémica y la posibilidad de supervivencia.

ABSTRACT Background: the acute mesenteric ischemia is a highly lethal disease, mainly due to the rapid evolution from its premature stage to the late one, the symptomatology in its start-up period and for the complexity of its management. Objective: to determine the clinical, laboratory, imagenological and evolutionary differences between the stages of the disease. Methods: an analytical transversal study was carried out. The universe was composed by 32 patients with acute mesenteric ischemia positive diagnosis in the period from December 2015 to March 2018, in Vladimir Ilich Lenin General Hospital. Results: peritoneal reaction is linked to the advanced stage in a highly significant way. There were no air-fluid levels in most of the ischemic patients, unlike the nechrosis stage, where levels were present in most of the patients. The difference between the pH and lactate average values in both of disease phases were statistically highly significant. All the cases with atrial fibrillation were dianosed in the late stage. All the patients with intestinal necrosis and more than 2/3 of the affected with intestinal ischemia perished. Conclusions: the delay in diagnosis was associated with high mortality. The early clinical suspicion of an acute abdomen, the assessment of hemodynamic parameters and the identification of risk factors such as atrial fibrillation, favor timely treatment in the ischemic stage and the possibility of survival.

Living Senior Labs, ecosistemas de co-creación e innovación abierta con personas mayores: revisión sistemática de la literatura en Ciencias Sociales / Living Senior Labs, co-criação e ecossistemas de inovação aberta com idosos: revisão sistemática da literatura em Ciências Sociais / Living Senior Labs, ecosystems of co-creation and open innovation with older people: a systematic review on Social Science literature

Los Living Labs son experiencias colaborativas que buscan implicar a la ciudadanía en la gobernanza científica y la evaluación de tecnologías. A pesar de su interés, se sabe muy poco sobre estas comunidades, su funcionamiento, tipología y características. Por ello, se realizó una revisión sistemática de la literatura sobre de un tipo particular de Living Lab, orientado hacia las personas mayores: los Living Senior Labs. A partir de una búsqueda general en las principales bases de datos científicas (WOS y Scopus), y de la aplicación de criterios de inclusión preestablecidos tras la primera selección quedaron finalmente seleccionados 19 estudios sobre Senior Labs (2010 y 2021). Los resultados proporcionan un mejor conocimiento de este tipo de ecosistemas y crean una base firme para avanzar en el conocimiento de este campo. (AU)

Os Living Labs são experiências colaborativas que procuram envolver os cidadãos na governança científica e na avaliação tecnológica. Apesar de seu interesse, muito pouco se sabe sobre essas comunidades, seu funcionamento, tipologia e características. Portanto, foi realizada uma revisão sistemática da literatura sobre um tipo particular de Living Lab, orientado para os idosos: os Living Senior Labs. Com base em uma pesquisa geral nos principais bancos de dados científicos (WOS e Scopus) e na aplicação de critérios de inclusão pré-estabelecidos, foram selecionados 19 estudos sobre os Senior Labs (2010 a 2021). Os resultados proporcionam uma melhor compreensão deste tipo de ecossistema e criam uma base firme para o avanço do conhecimento neste campo. (AU)

Living Labs are collaborative experiences that seek to involve citizens in scientific governance and technology assessment. In spite of their interest, very little is known about these communities, their functioning, typology and characteristics. Thus, a systematic literature review was carried out about a particular type of Living Lab, oriented towards the elderly: Living Senior Labs. Based on a general search in the main scientific databases (WOS and Scopus), and the application of pre-established inclusion criteria, 19 studies about Senior Labs were selected (2010 to 2021). The results provide a more comprehensive understanding of this type of ecosystem and create a strong foundation for progress in the knowledge of this area. (AU)

Management of Neuroinflammatory Responses to AAV-Mediated Gene Therapies for Neurodegenerative Diseases.

Recently, adeno-associated virus (AAV)-mediated gene therapies have attracted clinical interest for treating neurodegenerative diseases including spinal muscular atrophy (SMA), Canavan disease (CD), Parkinson's disease (PD), and Friedreich's ataxia (FA). The influx of clinical findings led to the first approved gene therapy for neurodegenerative disorders in 2019 and highlighted new safety concerns for patients. Large doses of systemically administered AAV stimulate host immune responses, resulting in anti-capsid and anti-transgene immunity with implications for transgene expression, treatment longevity, and patient safety. Delivering lower doses directly to the central nervous system (CNS) is a promising alternative, resulting in higher transgene expression with decreased immune responses. However, neuroinflammatory responses after CNS-targeted delivery of AAV are a critical concern. Reported signs of AAV-associated neuroinflammation in preclinical studies include dorsal root ganglion (DRG) and spinal cord pathology with mononuclear cell infiltration. In this review, we discuss ways to manage neuroinflammation, including choice of AAV capsid serotypes, CNS-targeting routes of delivery, genetic modifications to the vector and/or transgene, and adding immunosuppressive strategies to clinical protocols. As additional gene therapies for neurodegenerative diseases enter clinics, tracking biomarkers of neuroinflammation will be important for understanding the impact immune reactions can have on treatment safety and efficacy.

Evolutionarily unique mechanistic framework of clathrin-mediated endocytosis in plants.

In plants, clathrin mediated endocytosis (CME) represents the major route for cargo internalisation from the cell surface. It has been assumed to operate in an evolutionary conserved manner as in yeast and animals. Here we report characterisation of ultrastructure, dynamics and mechanisms of plant CME as allowed by our advancement in electron microscopy and quantitative live imaging techniques. Arabidopsis CME appears to follow the constant curvature model and the bona fide CME population generates vesicles of a predominantly hexagonal-basket type; larger and with faster kinetics than in other models. Contrary to the existing paradigm, actin is dispensable for CME events at the plasma membrane but plays a unique role in collecting endocytic vesicles, sorting of internalised cargos and directional endosome movement that itself actively promote CME events. Internalized vesicles display a strongly delayed and sequential uncoating. These unique features highlight the independent evolution of the plant CME mechanism during the autonomous rise of multicellularity in eukaryotes.

Long-Term Effects of Altered Photoperiod During Pregnancy on Liver Gene Expression of the Progeny.

Experimental and epidemiological studies have revealed a relationship between an adverse intrauterine environment and chronic non-communicable disease (NCD) like cardiovascular disease (CVD) in adulthood. An important risk factor for CVD is the deregulation of the fibrinolytic system particularly high levels of expression of plasminogen activator inhibitor 1 (Pai-1). Chronic exposure to altered photoperiod disrupts the circadian organization of physiology in the pregnant female, known as gestational chronodisruption, and cause long-term effects on the adult offspring's circadian physiology. The Pai-1 expression is regulated by the molecular components of the circadian system, termed clock genes. The present study aimed to evaluate the long-term effects of chronic photoperiod shifts (CPS) during pregnancy on the expression of the clock genes and the fibrinolytic system in the liver of adult male offspring. Our results using an animal model demonstrated statistically significant differences at the transcriptional level in males gestated under CPS. At 90 days of postnatal age, the liver transcript levels of the clock gene Bmal1 were downregulated, whereas Rorα, Rorγ, Nfil3, and Pai-1 were upregulated. Our data indicate that CPS during pregnancy affects gene expression in the liver of male adult progeny, showing that alteration of the photoperiod in the mother's environment leads to persistent effects in the offspring. In conclusion, these results reveal for the first time the long-term effects of gestational chronodisruption on the transcriptional activity of one well-established risk factor associated with CVD in the adult male offspring.