Creation of an Isogenic Human iPSC-Based RGC Model of Dominant Optic Atrophy Harboring the Pathogenic Variant c.1861C>T (p.Gln621Ter) in the OPA1 Gene.

Autosomal dominant optic atrophy (ADOA) is a rare progressive disease mainly caused by mutations in OPA1, a nuclear gene encoding for a mitochondrial protein that plays an essential role in mitochondrial dynamics, cell survival, oxidative phosphorylation, and mtDNA maintenance. ADOA is characterized by the degeneration of retinal ganglion cells (RGCs). This causes visual loss, which can lead to legal blindness in many cases. Nowadays, there is no effective treatment for ADOA. In this article, we have established an isogenic human RGC model for ADOA using iPSC technology and the genome editing tool CRISPR/Cas9 from a previously generated iPSC line of an ADOA plus patient harboring the pathogenic variant NM_015560.3: c.1861C>T (p.Gln621Ter) in heterozygosis in OPA1. To this end, a protocol based on supplementing the iPSC culture media with several small molecules and defined factors trying to mimic embryonic development has been employed. Subsequently, the created model was validated, confirming the presence of a defect of intergenomic communication, impaired mitochondrial respiration, and an increase in apoptosis and ROS generation. Finally, we propose the analysis of OPA1 expression by qPCR as an easy read-out method to carry out future drug screening studies using the created RGC model. In summary, this model provides a useful platform for further investigation of the underlying pathophysiological mechanisms of ADOA plus and for testing compounds with potential pharmacological action.

Plasma rich in growth factors versus corticosteroid injections for management of chronic rotator cuff tendinopathy: a prospective double-blind randomized controlled trial with 1 year of follow-up.

BACKGROUND: Rotator cuff tendinopathy (RCT) is a painful and dysfunctional shoulder condition traditionally considered as a degenerative pathology. However, evidence is pointing to immunocompetent cells and activated stromal fibroblasts as the drivers of a nonresolved inflammatory condition in RCT. As potent anti-inflammatory agents, corticosteroid injections have been among the first-line and the most common therapeutic strategies. Recently, another adjuvant therapy to treat musculoskeletal inflammation-driven painful conditions, namely, platelet-rich plasma (PRP), has emerged as safe and effective. The aim of this study was to compare the clinical efficacy of intratendinous injections of plasma rich in growth factors (PRGF) with conventional intratendinous corticosteroid injections on patients with chronic RCT using patient-reported outcome measures. METHODS: A total of 39 patients received PRGF treatment (3 infiltrations, 1 every other week), whereas 40 patients, as a control group, received corticosteroid (3 infiltrations, 1 every other week). Patients were evaluated before treatment and at 3, 6, and 12 months of follow-up using the University of California Los Angeles (UCLA) scale, Quick Disabilities of the Arm, Shoulder and Hand (QuickDASH), and Constant test. The primary outcome of the study was a 15% superior improvement of the PRGF group compared with the corticosteroid group in the UCLA scale and QuickDASH test at 6 months of follow-up, considering this difference to be clinically relevant. RESULTS: Both PRGF and corticosteroid groups showed significant clinical improvement in the 3 scores at all time points of the study compared with baseline. However, at 6 and 12 months of follow-up, the PRGF group had 22.1% and 21.2% superior improvement of the UCLA test, 14.3% and 13.5% for QuickDASH, and 16.4% and 20.2% for the Constant-Murley test, respectively, compared to the corticosteroid group. CONCLUSIONS: Three PRGF intratendinous injections every other week in patients with chronic rotator cuff tendinopathy show significantly superior and sustained pain-relieving and functional improvements compared with corticosteroid intratendinous injections assessed by 3 patient-reported outcome scales at 6 and 12 months of follow-up.

Anterior tibiofemoral dislocation after total knee arthroplasty: A case report and literature review.

Anterior tibiofemoral dislocation after total knee arthroplasty is an extremely rare and serious event. Amongst English-published papers, we found only 15 relevant cases, 3 of which presented vascular complications. This manuscript aims to present a 77-year-old woman with a TC-Plus (Smith & Nephew) cruciate-retaining type in first time of knee prosthesis, who suffered an anterior tibiofemoral dislocation and were admitted to our hospital. The clinical management and outcome were evaluated. Furthermore, a review of literature was performed. We concluded that early detection and surgical intervention of vascular injury is the key in the survival of the limbs. If there is still knee instability after acute recovery, it seems that revision surgery with constrained total knee arthroplasty can bring about good clinical and functional results.

Bread Feeding Is a Robust and More Physiological Enteropathogen Administration Method Compared to Oral Gavage.

Oral administration is a preferred model for studying infection by bacterial enteropathogens such as Yersinia spp. In the mouse model, the most frequent method for oral infection consists of oral gavage with a feeding needle directly introduced in the animal stomach via the esophagus. In this study, we compared needle gavage to bread feeding as an alternative mode of bacterial administration. Using bioluminescence-expressing strains of Yersinia pseudotuberculosis and Yersinia enterocolitica, we detected very early upon needle gavage a bioluminescent signal in the neck area together with a signal in the abdominal region, highlighting the presence of two independent sites of bacterial colonization and multiplication. Bacteria were often detected in the esophagus and trachea, as well as in the lymph nodes draining the salivary glands, suggesting that lesions made during needle introduction into the animal oral cavity lead to rapid bacterial draining to proximal lymph nodes. We then tested an alternative mode of bacterial administration using pieces of bread containing bacteria. Upon bread feeding infection, mice exhibited a stronger bioluminescent signal in the abdominal region than with needle gavage, and no signal was detected in the neck area. Moreover, Y. pseudotuberculosis incorporated in the bread is less susceptible to the acidic environment of the stomach and is therefore more efficient in causing intestinal infections. Based on our observations, bread feeding constitutes a natural and more efficient administration method which does not require specialized skills, is less traumatic for the animal, and results in diseases that more closely mimic foodborne intestinal infection.

iPSCs: A powerful tool for skeletal muscle tissue engineering.

Both volumetric muscle loss (VML) and muscle degenerative diseases lead to an important decrease in skeletal muscle mass, condition that nowadays lacks an optimal treatment. This issue has driven towards an increasing interest in new strategies in tissue engineering, an emerging field that can offer very promising approaches. In addition, the discovery of induced pluripotent stem cells (iPSCs) has completely revolutionized the actual view of personalized medicine, and their utilization in skeletal muscle tissue engineering could, undoubtedly, add myriad benefits. In this review, we want to provide a general vision of the basic aspects to consider when engineering skeletal muscle tissue using iPSCs. Specifically, we will focus on the three main pillars of tissue engineering: the scaffold designing, the selection of the ideal cell source and the addition of factors that can enhance the resemblance with the native tissue.

The Challenge of Bringing iPSCs to the Patient.

The implementation of induced pluripotent stem cells (iPSCs) in biomedical research more than a decade ago, resulted in a huge leap forward in the highly promising area of personalized medicine. Nowadays, we are even closer to the patient than ever. To date, there are multiple examples of iPSCs applications in clinical trials and drug screening. However, there are still many obstacles to overcome. In this review, we will focus our attention on the advantages of implementing induced pluripotent stem cells technology into the clinics but also commenting on all the current drawbacks that could hinder this promising path towards the patient.

Prevalence, genetic diversity and eco-epidemiology of pathogenic Leptospira species in small mammal communities in urban parks Lyon city, France.

Rodents are recognized as the main reservoirs of Leptospira spp. Rats, in particular, serve as hosts for the widely predominant Leptospira interrogans serovar Icterohaemorrhagiae, found worldwide. Several studies have shown the importance of other reservoirs, such as mice or hedgehogs, which harbor other leptospires' serovars. Nevertheless, our knowledge of circulating Leptospira spp. in reservoirs other than rats remains limited. In this context, we proposed an eco-health approach to assess the health hazard associated with leptospires in urban green spaces, where contacts between human/small mammals and domestic animals are likely. We studied the prevalence, the diversity of circulating strains, and epidemiology of pathogenic Leptospira species in small terrestrial mammal communities (rodents and shrews), between 2020-2022, in two parks in Lyon metropolis, France. Our study showed a significant carriage of Leptospira spp. in small terrestrial mammals in these parks and unveiled a global prevalence rate of 11.4%. Significant variations of prevalence were observed among the small mammal species (from 0 to 26.1%), with Rattus norvegicus exhibiting the highest infection levels (26.1%). We also observed strong spatio-temporal variations in Leptospira spp. circulation in its reservoirs. Prevalence seems to be higher in the peri-urban park and in autumn in 2021 and 2022. This is potentially due to differences in landscape, abiotic conditions and small mammal communities' composition. Our study suggests an important public health relevance of rats and in a lesser extent of other rodents (Apodemus spp., Clethrionomys glareolus and Mus musculus) as reservoirs of L. interrogans, with rodent species carrying specific serogroups/serovars. We also emphasize the potential hazard associated between the shrew Crocidura russula and L. kirschneri. Altogether, these results improve our knowledge about the prevalence of leptospirosis in an urban environment, which is an essential prerequisite for the implementation of prevention of associated risks.

Genetic diversity of Leptospira strains circulating in humans and dogs in France in 2019-2021.

Leptospirosis is a bacterial zoonotic disease. Humans and dogs are susceptible hosts, with similar clinical manifestations ranging from a febrile phase to multiple organ dysfunction. The incidence of leptospirosis in mainland France is relatively high, at about 1 case per 100,000 inhabitants, but our knowledge of the strains circulating in humans and dogs remains limited. We studied the polymorphism of the lfb1 gene sequences in an exhaustive database, to facilitate the identification of Leptospira strains. We identified 46 species-groups (SG) encompassing the eight pathogenic species of Leptospira. We sequenced the lfb1 gene amplification products from 170 biological samples collected from 2019 to 2021: 110 from humans and 60 from dogs. Epidemiological data, including vaccination status in dogs, were also collected. Three Leptospira species displaying considerable diversity were identified: L. interrogans, with eight lfb1 species-groups (including five new lfb1 species-groups) in humans and dogs; L. kirschneri, with two lfb1 species-groups in humans and dogs; and L. borgpetersenii, with one lfb1 species-group in humans only. The lfb1 species-group L. interrogans SG1, corresponding to serovar Icterohaemorrhagiae or Copenhageni, was frequently retrieved from both humans and dogs (n=67/110; 60.9% and n=59/60; 98.3% respectively). A high proportion of the affected dogs developed the disease despite vaccination (n=30/60; 50%). Genotyping with the polymorphic lfb1 gene is both robust and simple. This approach provided the first global picture of the Leptospira strains responsible for acute infections in mainland France, based on biological samples but without the need for culture. Identification of the Leptospira strains circulating and their changes over time will facilitate more precise epidemiological monitoring of susceptible and reservoir species. It should also facilitate the monitoring of environmental contamination, making it possible to implement preventive measures and to reduce the burden of this disease.

Gallocin A, an Atypical Two-Peptide Bacteriocin with Intramolecular Disulfide Bonds Required for Activity.

Streptococcus gallolyticus subsp. gallolyticus (SGG) is an opportunistic gut pathogen associated with colorectal cancer. We previously showed that colonization of the murine colon by SGG in tumoral conditions was strongly enhanced by the production of gallocin A, a two-peptide bacteriocin. Here, we aimed to characterize the mechanisms of its action and resistance. Using a genetic approach, we demonstrated that gallocin A is composed of two peptides, GllA1 and GllA2, which are inactive alone and act together to kill "target" bacteria. We showed that gallocin A can kill phylogenetically close relatives of the pathogen. Importantly, we demonstrated that gallocin A peptides can insert themselves into membranes and permeabilize lipid bilayer vesicles. Next, we showed that the third gene of the gallocin A operon, gip, is necessary and sufficient to confer immunity to gallocin A. Structural modeling of GllA1 and GllA2 mature peptides suggested that both peptides form alpha-helical hairpins stabilized by intramolecular disulfide bridges. The presence of a disulfide bond in GllA1 and GllA2 was confirmed experimentally. Addition of disulfide-reducing agents abrogated gallocin A activity. Likewise, deletion of a gene encoding a surface protein with a thioredoxin-like domain impaired the ability of gallocin A to kill Enterococcus faecalis. Structural modeling of GIP revealed a hairpin-like structure strongly resembling those of the GllA1 and GllA2 mature peptides, suggesting a mechanism of immunity by competition with GllA1/2. Finally, identification of other class IIb bacteriocins exhibiting a similar alpha-helical hairpin fold stabilized with an intramolecular disulfide bridge suggests the existence of a new subclass of class IIb bacteriocins. IMPORTANCE Streptococcus gallolyticus subsp. gallolyticus (SGG), previously named Streptococcus bovis biotype I, is an opportunistic pathogen responsible for invasive infections (septicemia, endocarditis) in elderly people and is often associated with colon tumors. SGG is one of the first bacteria to be associated with the occurrence of colorectal cancer in humans. Previously, we showed that tumor-associated conditions in the colon provide SGG with an ideal environment to proliferate at the expense of phylogenetically and metabolically closely related commensal bacteria such as enterococci (1). SGG takes advantage of CRC-associated conditions to outcompete and substitute commensal members of the gut microbiota using a specific bacteriocin named gallocin, recently renamed gallocin A following the discovery of gallocin D in a peculiar SGG isolate. Here, we showed that gallocin A is a two-peptide bacteriocin and that both GllA1 and GllA2 peptides are required for antimicrobial activity. Gallocin A was shown to permeabilize bacterial membranes and kill phylogenetically closely related bacteria such as most streptococci, lactococci, and enterococci, probably through membrane pore formation. GllA1 and GllA2 secreted peptides are unusually long (42 and 60 amino acids long) and have very few charged amino acids compared to well-known class IIb bacteriocins. In silico modeling revealed that both GllA1 and GllA2 exhibit a similar hairpin-like conformation stabilized by an intramolecular disulfide bond. We also showed that the GIP immunity peptide forms a hairpin-like structure similar to GllA1/GllA2. Thus, we hypothesize that GIP blocks the formation of the GllA1/GllA2 complex by interacting with GllA1 or GllA2. Gallocin A may constitute the first class IIb bacteriocin which displays disulfide bridges important for its structure and activity and might be the founding member of a subtype of class IIb bacteriocins.

Hereditary Optic Neuropathies: Induced Pluripotent Stem Cell-Based 2D/3D Approaches.

Inherited optic neuropathies share visual impairment due to the degeneration of retinal ganglion cells (RGCs) as the hallmark of the disease. This group of genetic disorders are caused by mutations in nuclear genes or in the mitochondrial DNA (mtDNA). An impaired mitochondrial function is the underlying mechanism of these diseases. Currently, optic neuropathies lack an effective treatment, and the implementation of induced pluripotent stem cell (iPSC) technology would entail a huge step forward. The generation of iPSC-derived RGCs would allow faithfully modeling these disorders, and these RGCs would represent an appealing platform for drug screening as well, paving the way for a proper therapy. Here, we review the ongoing two-dimensional (2D) and three-dimensional (3D) approaches based on iPSCs and their applications, taking into account the more innovative technologies, which include tissue engineering or microfluidics.

Generation of the iPSC line IISHDOi007-A from peripheral blood mononuclear cells from a patient with McArdle disease harbouring the mutation c.2392 T > C; p.Trp798Arg.

Peripheral blood mononuclear cells (PBMCs) from a McArdle patient carrying a homozygous mutation in the PYGM gene: c.2392 T > C; p.Trp798Arg were used for the generation of the human iPSC line, IISHDOi007-A. For the delivery of the reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc, a non-integrative methodology that implies the use of Sendai virus has been applied.

Derivation of an aged mouse induced pluripotent stem cell line, IISHDOi005-A.

A mouse iPSC line, IISHDOi005-A, generated from fibroblasts obtained from a mouse C57BL/6J with an age of 1 year and a half, has been obtained. For this purpose, reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using Sendai virus.

Derivation of a human DOA iPSC line, IISHDOi006-A, with a mutation in the ACO2 gene: c.1999G>A; p.Glu667Lys.

Human iPSC line, IISHDOi006-A, was obtained from fibroblasts of a patient with Dominant Optic Atrophy (DOA) carrying a heterozygous mutation in the gene ACO2: c.1999G>A; p.Glu667Lys. Reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using a non-integrative methodology that involves the use of Sendai virus.

Establishment of a human iPSC line (IISHDOi001-A) from a patient with McArdle disease.

Human iPSC line IISHDOi001-A was generated from fibroblasts of a patient with McArdle disease harbouring the mutation, c.148C>T; p.Arg50Ter, in the PYGM gene. Reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using Sendai virus.

Estado nutricional y hábitos alimenticios en un equipo de fútbol profesional / Nutritional status and feeding habits in a professional soccer team

Pese a que el fútbol es el deporte más popular del planeta, la nutrición del futbolista está ciertamente descuidada. La actividad física exige una utilización mixta de los sistemas fisiológicos de obtención de energía, aeróbico y anaeróbico, requiriéndose una completa ingesta de nutrientes; siendo de sobra conocido el papel de los carbohidratos en el rendimiento deportivo. OBJETIVOS: El objetivo de este estudio es determinar la ingesta nutricional de macronutrientes en un equipo de fútbol profesional. MATERIAL Y MÉTODOS: Realizamos un estudio epidemiológico, descriptivo y prospectivo de la primera plantilla de un equipo de la Segunda División B española. Se trata de 22 jugadores masculinos, con edad media de 26,19 (19,5-31,6). Elaboramos un protocolo de recogida de alimentos ingeridos durante 7 días, contabilizando la ingesta de macronutrientes y cuantificándose el gasto energético total, variable en función del día de entrenamiento y posición del jugador. RESULTADOS: El total de carbohidratos globales consumidos fue de 305,07 +/- 56 gr (48,78% de las calorías). Estos valores varían entre el día de actividad moderada: 213,6 +/- 37 gr (39,93%) y el día de actividad intensa: 361,84 +/- 28 gr (49,21%). Existen diferencias significativas en el total de carbohidratos según la actividad diaria. Además, encontramos diferencias significativas en la altura del jugador, su tasa metabólica basal y gasto energético total, entre las posiciones de portero y jugador de banda avanzado (p < 0,05). DISCUSIÓN Y CONCLUSIONES: Tanto el aporte de carbohidratos como la cantidad global de calorías consumidas por el futbolista se encuentran por debajo del estándar recomendado (3600 kcal/día)

Despite soccer is the most popular sport on the planet, soccer player nutrition is certainly neglected. Physical activity requires a mixed use of the physiological systems for obtaining energy, requiring a complete intake of nutrients; and the role of carbohydrates in sports performance is well known. OBJECTIVES: The objective of this study is to determine the nutritional intake of macronutrients in a professional soccer team. MATERIAL AND METHODS: We carried out an epidemiological, descriptive and prospective study of a Spanish Second Division B team. They were 22 male players, with a mean age of 26.19 (19.5-31.6). We elaborated a protocol for collecting food eaten for 7 days, counting the macronutrient intake and quantifying the total energy expenditure, which varies depending on the training day and the player's position. RESULTS: The global carbohydrates consumed was 305.07+/- 56 gr (48.78% of calories). These values vary between the moderate activity day: 213.6 +/- 37 grams (39.93%) and the intense activity day: 361.84 +/- 28 grams (49.21%). We found significant differences in total carbohydrates based on daily activity. In addition, we found significant differences in the player height, their basal metabolic rate and total energy expenditure, between the positions of goal keeper and advanced band player (p <0.05). DISCUSSION AND CONCLUSIONS: Both carbohydrate intake and the overall amount of calories consumed by the footballer are below the recommended standard (3600 kcal / day)

Referencias para dinamometría manual en función de la estatura en edad pediátrica y adolescente / Hand grip dynamometry references by height ranges in pediatric and adolescent age

Antecedentes y objetivo: La dinamometría de la mano es una prueba funcional recomendada en la evaluación nutricional. Suele expresarse en función de la edad pero durante el crecimiento, incluso a la misma edad, varía el tamaño corporal. El objetivo del presente trabajo es aportar referencias de la fuerza de la mano en función de la talla. Sujetos y método: La muestra consta de 1.798 escolares entre 6 y 15 años. Se midió la talla, el peso y la fuerza de ambas manos. La dinamometría media (DM) se expresó en función de la estatura y se obtuvieron ecuaciones predictivas. La estadística descriptiva, comparativa y el análisis de regresión se efectuaron con el SPSS 21.0 Resultados: Se aportan referencias percentilares para la DM por categorías de talla en rangos de 5 cm. Las fórmulas obtenidas a partir del modelo de regresión (varones: DM = 0,207*peso + 0,621*talla - 71,461; mujeres DM=0,258*peso + 0,394*talla - 43,967) fueron validadas con la mitad de la muestra. Conclusiones: Las ecuaciones predictivas que se aportan en este trabajo permiten estimar la DM a partir del peso y la estatura con independencia de la edad (AU)

Background and objectives: Hand grip dynamometry is a functional test recommended in the nutritional assessment. Usually it is expressed in terms of age but during growth even at the same age, body size varies. The aim of this study is to provide references of hand strength depending on the height. Subjects and method: The sample consists of 1,798 students between 6 and 15 years. Height, weight and strength of both hands were measured. The mean dynamometry (MD) was expressed in terms of height and predictive equations were obtained. Descriptive statistics, comparative and regression analysis were performed using SPSS 21.0 Results: Percentile references for MD are provided by height categories in ranges of 5 cm. The obtained formulas from the regression model (males: MD = 0.207 * 0.621 * weight + size - 71.461; females: MD = 0.258 * 0.394 * weight + size - 43.967) were validated in half of the sample. Conclusions: The predictive equations given in this paper allow estimating MD from weight and height independently of age (AU)