Ocular motor nerve palsy in patients with diabetes: High-resolution MR imaging of nerve enhancement.

OBJECTIVE: To evaluate the extent of signal abnormality in impaired ocular motor nerves using high signal and spatial resolution MRI sequences and to discuss the involvement of inflammatory or microvascular impairment in patients with diabetic ophthalmoplegia. METHODS: We conducted a retrospective study of 10 patients referred for acute ocular motor nerve palsy in the context of diabetes mellitus from September 15th, 2021 to April 24th, 2022. 3T MRI evaluation included diffusion, 3D TOF, FLAIR, coronal STIR and post-injection 3D T1 SPACE DANTE sequences. RESULTS: Ten patients were included: 9 males and 1 female aged from 46 to 79 years. Five patients presented with cranial nerve (CN) III palsy, and 5 presented with CN VI palsy. Third nerve palsy was pupil-sparing in 4 patients and pupil-involved in 1 patient. Pain was associated in all patients with CN III deficiencies and in 2 patients CN VI deficiencies. In all patients, MRI sequences ruled out mass effect and vascular pathology, such as acute stroke or aneurysm. Eight patients presented with STIR hypersignals, some with enlargement of the involved nerve. The diagnosis was confirmed through a post-injection 3D T1 SPACE DANTE sequence, which showed extended enhancement along the abnormal portion of the nerve. CONCLUSION: High-resolution MRI evaluation of diplopia in diabetic patients is used to rule out a diagnosis of acute stroke and contributes to the positive diagnosis of ocular motor nerve impairment, possibly combining the influences of inflammatory and microvascular phenomena. Dedicated MR imaging should be included in the initial diagnosis and longitudinal follow-up of patients with diabetic ophthalmoplegia.

The Association between Metabolic Syndrome, Frailty and Disability-Free Survival in Healthy Community-dwelling Older Adults.

OBJECTIVES: To examine the association between metabolic syndrome (MetS) and frailty, and determine whether co-existent MetS and frailty affect disability-free survival (DFS), assessed through a composite of death, dementia or physical disability. DESIGN: Longitudinal study. SETTING AND PARTICIPANTS: Community-dwelling older adults from Australia and the United States (n=18,264) from "ASPirin in Reducing Events in the Elderly" (ASPREE) study. MEASUREMENTS: MetS was defined according to American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines (2018). A modified Fried phenotype and a deficit accumulation Frailty Index (FI) were used to assess frailty. Association between MetS and frailty was examined using multinomial logistic regression. Cox regression was used to analyze the association between MetS, frailty and DFS over a median follow-up of 4.7 years. RESULTS: Among 18,264 participants, 49.9% met the criteria for MetS at baseline. Participants with Mets were more likely to be pre-frail [Relative Risk Ratio (RRR): 1.22; 95%Confidence Interval (CI): 1.14, 1.30)] or frail (RRR: 1.66; 95%CI: 1.32, 2.08) than those without MetS. MetS alone did not shorten DFS while pre-frailty or frailty alone did [Hazard Ratio (HR): 1.68; 95%CI: 1.45, 1.94; HR: 2.65; 95%CI:1.92, 3.66, respectively]. Co-existent MetS with pre-frailty/frailty did not change the risk of shortened DFS. CONCLUSIONS: MetS was associated with pre-frailty or frailty in community-dwelling older individuals. Pre-frailty or frailty increased the risk of reduced DFS but presence of MetS did not change this risk. Assessment of frailty may be more important than MetS in predicting survival free of dementia or physical disability.

Adaptación transcultural del cuestionario motion sickness susceptibility questionnaire form short (MSSQ-SHORT) para la población adulta chilena / Cross-cultural adaptation of the motion sickness susceptibility questionnaire form short (MSSQ-SHORT) for the chilean adult population

Resumen Introducción: La cinetosis se relaciona con la presencia de una serie de síntomas que comúnmente son inducidos por situaciones cotidianas de viajes en medios de transporte. Una forma utilizada por décadas para determinar el grado de susceptibilidad a la cinetosis ha sido con la aplicación del cuestionario en su versión acortada Motion Sickness Suscep-tibility-short (MSSQ-short). Objetivo: Adaptar lingüística y transculturalmente al español el cuestionario MSSQ-short. Material y Método: Se llevaron a cabo cuatro etapas: Traducción directa, traducción inversa (retrotraducción), consolidación por un comité de expertos y pretest (aplicabilidad/viabilidad). En la etapa de pre-test 51 personas respondieron el cuestionario. Resultados: La discrepancias encontradas en las primeras etapas fueron resueltas por un tercer traductor, el cual concluyó en un documento final en español que fue analizado y revisado por el comité de expertos. Se determinaron los percentiles del 0 al 100, percentil 50 con 9,0 puntos, percentil 25 con 2,13 puntos y el percentil 75 con 17,4 puntos. La consistencia interna del cuestionario fue de 0,889. Conclusión: La traducción y adaptación transcultural fue aceptada por un comité de expertos y participantes con distintas características demográficas y educacionales. El cuestionario obtuvo buena consistencia interna y resultados concordantes con la versión original.

Abstract Introduction: Motion sickness is related to the presence of a series of symptoms that are typically induced by everyday situations of travel in means of transport. A way used for decades to determine the degree of susceptibility to motion sickness has been with the application of the questionnaire in its shortened version Motion Sickness Susceptibility-short (MSSQ-short). Aim: Linguistically and cross-culturally adapt the MSSQ-short questionnaire to Spanish. Material and Method: Four stages were carried out: direct translation, reverse translation (back translation), consolidation by a committee of experts, and pretest (applicability/feasibility). In the pre-test stage, 51 people answered the questionnaire. Results: The discrepancies found in the early stages were resolved by a third translator, which concluded in a final document in Spanish that was analyzed and reviewed by the expert committee. The percentiles from 0 to 100 were determined, 50th percentile with 9.0 points, 25th percentile with 2.13 points, and 75th percentile with 17.4 points. The internal consistency of the questionnaire was 0.889. Conclusion: The cross-cultural translation and adaptation were accepted by a committee of experts and participants with different demographic and educational characteristics. The questionnaire obtained good internal consistency and results consistent with the original version.

A Behavioral Lifestyle Intervention to Improve Frailty in Overweight or Obese Older Adults with Type 2 Diabetes: A Feasibility Study.

BACKGROUND: Older adults with Type 2 diabetes (T2D) are more likely to be frail, which increases the risk for disability and mortality. OBJECTIVES: To determine the feasibility of a behavioral lifestyle intervention, enhanced with mobile health technology for self-monitoring of diet and activity, to improve frailty in overweight/obese older adults (≥65 years) diagnosed with T2D. DESIGN, SETTING, AND PARTICIPANTS: Single arm, 6-month study of a behavioral lifestyle intervention in 20 overweight/obese (BMI>25) older adults (≥ 65 years) with self-reported T2D diagnosis who owned a smartphone. A Fitbit tracker was provided to all participants for self-monitoring of diet and physical activity. Our primary outcome of feasibility was measured by session attendance, adherence to Fitbit usage to self-monitor diet and physical activity, and study retention. Secondary outcomes included the preliminary efficacy of the intervention on frailty, physical function, quality of life, and T2D-related outcomes. RESULTS: Eighteen participants completed the study. The mean age was 71.5 (SD ± 5.3) years, 56% were female, and half were Hispanic. At baseline, 13 (72%) were pre-frail, 4 (22%) were frail, and 1 (6%) were non-frail. At follow-up, frailty scores improved significantly from 1.61 ± 1.15 to 0.94 ± 0.94 (p=0.01) and bodyweight improved from 205.66 ± 45.52 lbs. to 198.33 ± 43.6 lbs. (p=<0.001). CONCLUSION: This study provides evidence for the feasibility of a behavioral lifestyle intervention in overweight/obese older adults with T2D and preliminary results support its potential efficacy in improving frailty score.

Persistent epiphora after endonasal ENT surgery: CT dacryography findings.

OBJECTIVES: To analyze postoperative CT dacryography features in patients with persistent epiphora after endonasal surgery. METHODS: We conducted a retrospective study of 76 patients with a history of persistent epiphora after endonasal ENT surgery who underwent CT dacryography between January 2014 and February 2020. Volume acquisition of sub-millimeter sections allowed 2D and 3D reconstructions with virtual endoscopy of the nasosinusal cavities and the lacrimal canal. RESULTS: The postsurgical appearance of the nasosinusal cavities revealed a middle meatal antrostomy in 37% of cases, less frequently an ethmoidectomy or an inferior meatal antrostomy, sometimes completed by a middle or inferior turbinectomy. In thirty-five patients (46%), the lacrimal canal was distant from the endonasal ENT procedure. Epiphora was related to mucosal hypertrophy, constricting all or part of the lacrimal canal. Thirty-three patients (43%) showed changes in the lacrimal canal at the surgical site. In the inferior meatus, the nasolacrimal orifice was sometimes involved in the inferior turbinectomy or meatotomy, but most of the time, in the middle meatus, resection of the uncinate process prior to ethmoidectomy or middle meatotomy was associated with a lesion of the contiguous lacrimal canal. CONCLUSION: As a rare cause of persistent tearing, involvement of the nasolacrimal duct at the edge of the endonasal ENT surgery highlights the importance of intraoperative localization of the nasolacrimal duct before resection of the uncinate process or the inferior turbinate, ideally predicted by preoperative CT imaging.

Actin protein inside DMPC GUVs and its mechanical response to AC electric fields.

Cells are dynamic systems with complex mechanical properties, regulated by the presence of different species of proteins capable to assemble (and disassemble) into filamentous forms as required by different cells functions. Giant unilamellar vesicles (GUVs) of DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) are systems frequently used as a simplified model of cells because they offer the possibility of assaying separately different stimuli, which is no possible in living cells. Here we present a study of the effect of acting protein on mechanical properties of GUVs, when the protein is inside the vesicles in either monomeric G-actin or filamentous F-actin. For this, rabbit skeletal muscle G-actin is introduced inside GUVs by the electroformation method. Protein polymerization inside the GUVs is promoted by adding to the solution MgCl2 and the ion carrier A23187 to allow the transport of Mg+2 ions into the GUVs. To determine how the presence of actin changes the mechanical properties of GUVs, the vesicles are deformed by the application of an AC electric field in both cases with G-actin and with polymerized F-actin. The changes in shape of the vesicles are characterized by optical microscopy and from them the bending stiffness of the membrane are determined. It is found that G-actin has no appreciable effect on the bending stiffness of DMPC GUVs, but the polymerized actin makes the vesicles more rigid and therefore more resistant to deformations. This result is supported by evidence that actin filaments tend to accumulate near the membrane.

Particle/wall electroviscous effects at the micron scale: comparison between experiments, analytical and numerical models.

We report a experimental study of the motion of 1 µm single particles interacting with functionalized walls at low and moderate ionic strengths conditions. The 3D particle's trajectories were obtained by analyzing the diffracted particle images (point spread function). The studied particle/wall systems include negatively charged particles interacting with bare glass, glass covered with polyelectrolytes and glass covered with a lipid monolayer. In the low salt regime (pure water) we observed a retardation effect of the short-time diffusion coefficients when the particle interacts with a negatively charged wall; this effect is more severe in the perpendicular than in the lateral component. The decrease of the diffusion as a function of the particle-wall distancehwas similar regardless the origin of the negative charge at the wall. When surface charge was screened or salt was added to the medium (10 mM), the diffusivity curves recover the classical hydrodynamic behavior. Electroviscous theory based on the thin electrical double layer (EDL) approximation reproduces the experimental data except for smallh. On the other hand, 2D numerical solutions of the electrokinetic equations showed good qualitative agreement with experiments. The numerical model also showed that the hydrodynamic and Maxwellian part of the electroviscous total drag tend to zero ash→ 0 and how this is linked with the merging of both EDL's at close proximity.

Effects of DC Magnetic Fields on Magnetoliposomes.

The potential use of magnetic nanoparticles (MNPs) in biomedicine as magnetic resonance, drug delivery, imagenology, hyperthermia, biosensors, and biological separation has been studied in different laboratories. One of the challenges on MNP elaboration for biological applications is the size, biocompatibility, heat efficiency, stabilization in physiological conditions, and surface coating. Magnetoliposome (ML), a lipid bilayer of phospholipids encapsulating MNPs, is a system used to reduce toxicity. Encapsulated MNPs can be used as a potential drug and a gene delivery system, and in the presence of magnetic fields, MLs can be accumulated in a target tissue by a strong gradient magnetic field. Here, we present a study of the effects of DC magnetic fields on encapsulated MNPs inside liposomes. Despite their widespread applications in biotechnology and environmental, biomedical, and materials science, the effects of magnetic fields on MLs are unclear. We use a modified coprecipitation method to synthesize superparamagnetic nanoparticles (SNPs) in aqueous solutions. The SNPs are encapsulated inside phospholipid liposomes to study the interaction between phospholipids and SNPs. Material characterization of SNPs reveals round-shaped nanoparticles with an average size of 12 nm, mainly magnetite. MLs were prepared by the rehydration method. After formation, we found two types of MLs: one type is tense with SNPs encapsulated and the other is a floppy vesicle that does not show the presence of SNPs. To study the response of MLs to an applied DC magnetic field, we used a homemade chamber. Digitalized images show encapsulated SNPs assembled in chain formation when a DC magnetic field is applied. When the magnetic field is switched off, it completely disperses SNPs. Floppy MLs deform along the direction of the external applied magnetic field. Solving the relevant magnetostatic equations, we present a theoretical model to explain the ML deformations by analyzing the forces exerted by the magnetic field over the surface of the spheroidal liposome. Tangential magnetic forces acting on the ML surface result in a press force deforming MLs. The type of deformations will depend on the magnetic properties of the mediums inside and outside the MLs. The model predicts a coexistence region of oblate-prolate deformation in the zone where χ = 1. We can understand the chain formation in terms of a dipole-dipole interaction of SNP.

The Association between Frailty and All-Cause Mortality in Community-Dwelling Older Individuals: An Umbrella Review.

Frailty is associated with multiple adverse health outcomes, including mortality. Several methods have been used to characterize frailty, each based on different frailty scales. These include scales based on phenotype, multidomain, and deficit accumulations. Several systematic reviews have examined the association between frailty and mortality; however, it is unclear whether these different frailty scales similarly predict mortality. This umbrella review aims to examine the association between frailty assessed by different frailty scales and all-cause mortality among community-dwelling older adults. A protocol was registered at PROSPERO, and it was conducted following the PRISMA statement. MEDLINE, Embase, PubMed, Cochrane Database of Systematic Reviews, Joanna Briggs Institute (JBI) EBP database, and Web of Science database was searched. Methodological quality was assessed using the JBI critical appraisal checklist and online AMSTAR-2 critical appraisal checklist. For eligible studies, essential information was extracted and synthesized qualitatively. Five systematic reviews were included, with a total of 434,115 participants. Three systematic reviews focused on single frailty scales; one evaluated Fried's physical frailty phenotype and its modifications; another focused on the deficit accumulation frailty index. The third evaluated the FRAIL (Fatigue, Resistance, Ambulation, Illness, and Loss of weight) scale. The two other systematic reviews determined the association between frailty and mortality using different frailty scales. All of the systematic reviews found that frailty was significantly associated with all-cause mortality. This umbrella review demonstrates that frailty is a significant predictor of all-cause mortality, irrespective of the specific frailty scale.

Desafío diagnóstico y terapéutico de carcinoma mucoepidermoide palatino: reporte de un caso

Resumen Las neoplasias en glándulas salivales son infrecuentes, representando menos del 3% de los tumores de cabeza y cuello. El carcinoma mucoepidermoide es el tumor maligno más común en glándulas salivales, siendo su principal ubicación la parótida. Clínicamente se asemeja a otras lesiones de mucosa oral, por lo cual, es importante realizar un correcto diagnóstico diferencial. Su comportamiento biológico se relaciona con el grado histológico tumoral, factor relevante en el pronóstico y tratamiento de esta neoplasia. Presentamos el caso de un paciente hombre de 75 años afectado con un tumor en paladar con diagnóstico de carcinoma mucoepidermoide de bajo grado. Como tratamiento se realizó una maxilectomía parcial y una placa obturadora en base a una prótesis removible y posterior reconstrucción con un colgajo libre microvascularizado. Actualmente el paciente se encuentra en controles periódicos, libre de enfermedad. Los tumores de glándulas salivales son un desafío diagnóstico, requieren de exámenes imagenológicos y del estudio histopatológico. Cuando existen dudas en el diagnóstico, se debe considerar repetir la toma de la muestra o la obtención de biopsias de más de una zona representativa que permita el diagnóstico de la lesión.

Abstract Salivary gland neoplasms are infrequent lesions representing less than 3% of head and neck tumors. Mucoepidermoid carcinoma is the most common malignant tumor in salivary glands, being the parotid the most usual location. Clinically, it resembles other oral mucosa lesions, therefore, it is important to make a correct differential diagnosis. Its biological behavior is related to the tumor histological grade, a relevant factor in the prognosis and treatment of this neoplasm. We reported a case of a 75-year-old-man, with a tumor in the palate, diagnosed as low-grade mucoepidermoid carcinoma. A partial maxillectomy and an obturator plate were performed based on a removable prosthesis and subsequent reconstruction with a microvascularized free flap. The patient is currently undergoing regular checkups, maintaining disease free. Salivary gland tumors are a diagnostic challenge, requiring imaging tests and histopathological study. In case of doubts with the diagnosis, it should be considered to biopsy more than area or to repeat the biopsy in order to obtain a representative sample that allows the diagnosis of the lesion.

An Individualized Low-Intensity Walking Clinic Leads to Improvement in Frailty Characteristics in Older Veterans.

BACKGROUND: Sedentary lifestyle leads to worse health outcomes with aging, including frailty. Older adults can benefit from regular physical activity, but exercise promotion in the clinical setting is challenging. OBJECTIVES: The objective of this clinical demonstration project was to implement a Geriatric Walking Clinic for older adults and determine whether this clinical program can lead to improvements in characteristics of frailty. DESIGN: This was a clinical demonstration project/quality improvement project. SETTING: Outpatient geriatrics clinic at the South Texas Veterans Health Care System (STVHCS). PARTICIPANTS: Older Veterans, aged ≥60 years. INTERVENTION: A 6-week structured walking program, delivered by a registered nurse and geriatrician. Patients received a pedometer and a comprehensive safety evaluation at an initial face-to-face visit. They were subsequently followed with weekly phone calls and participated in a final face-to-face follow-up visit at 6 weeks. MEASUREMENTS: Grip strength (handheld dynamometer), gait speed (10-ft walk), Timed Up and Go (TUG), and body mass index (BMI) were assessed at baseline and follow-up. Frailty status for gait speed was assessed using Fried criteria. RESULTS: One hundred eighty five patients completed the program (mean age: 68.4 ±7 years, 88% male). Improvements from baseline to follow-up were observed in average steps/day, gait speed, TUG, and BMI. Improvement in gait speed (1.13 ±0.20 vs. 1.24 ± 0.23 meter/second, p<0.0001) resulted in reduced odds of meeting frailty criteria for slow gait at follow-up compared to the baseline examination (odds ratio = 0.31, 95% confidence interval: 0.13-0.72, p = 0.01). CONCLUSIONS: Our findings demonstrate that a short duration, low-intensity walking intervention improves gait speed and TUG. This new clinical model may be useful for the promotion of physical activity, and for the prevention or amelioration of frailty characteristics in older adults.

Network approach identifies Pacer as an autophagy protein involved in ALS pathogenesis.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a multifactorial fatal motoneuron disease without a cure. Ten percent of ALS cases can be pointed to a clear genetic cause, while the remaining 90% is classified as sporadic. Our study was aimed to uncover new connections within the ALS network through a bioinformatic approach, by which we identified C13orf18, recently named Pacer, as a new component of the autophagic machinery and potentially involved in ALS pathogenesis. METHODS: Initially, we identified Pacer using a network-based bioinformatic analysis. Expression of Pacer was then investigated in vivo using spinal cord tissue from two ALS mouse models (SOD1G93A and TDP43A315T) and sporadic ALS patients. Mechanistic studies were performed in cell culture using the mouse motoneuron cell line NSC34. Loss of function of Pacer was achieved by knockdown using short-hairpin constructs. The effect of Pacer repression was investigated in the context of autophagy, SOD1 aggregation, and neuronal death. RESULTS: Using an unbiased network-based approach, we integrated all available ALS data to identify new functional interactions involved in ALS pathogenesis. We found that Pacer associates to an ALS-specific subnetwork composed of components of the autophagy pathway, one of the main cellular processes affected in the disease. Interestingly, we found that Pacer levels are significantly reduced in spinal cord tissue from sporadic ALS patients and in tissues from two ALS mouse models. In vitro, Pacer deficiency lead to impaired autophagy and accumulation of ALS-associated protein aggregates, which correlated with the induction of cell death. CONCLUSIONS: This study, therefore, identifies Pacer as a new regulator of proteostasis associated with ALS pathology.

Identifying Exosome-Derived MicroRNAs as Candidate Biomarkers of Frailty.

Frailty is a geriatric syndrome associated with progressive physical decline and significantly increases risk for falls, disability, hospitalizations, and death. However, much remains unknown regarding the biological mechanisms that contribute to aging and frailty, and to date, there are no clinically used prognostic or diagnostic molecular biomarkers. The present study profiled exosome-derived microRNAs isolated from the plasma of young, robust older, and frail older individuals and identified eight miRNAs that are uniquely enriched in frailty: miR-10a-3p, miR-92a-3p, miR-185-3p, miR-194-5p, miR-326, miR-532-5p, miR-576-5p, and miR-760. Furthermore, since exosomes can deliver miRNAs to alter cellular activity and behavior, these miRNAs may also provide insights into the biological mechanisms underlying frailty; KEGG analysis of their target genes revealed multiple pathways implicated in aging and age-related processes. Although further validation and research studies are warranted, our study identified eight novel candidate biomarkers of frailty that may help to elucidate the multifactorial pathogenesis of frailty.

Increased dietary protein or free amino acids supply for heat stress pigs: effect on performance and carcass traits.

Heat stress (HS) pigs reduce their voluntary feed intake (VFI) and ingestion of indispensable amino acids (AA). Increasing the dietary crude protein (CP) content may help to correct the reduced AA intake by HS pigs, but it may further increase their body heat load. Increasing the AA intake by adding free AA to the diet does not affect the heat load of HS pigs. Two 21-d experiments were conducted. In Exp. 1, 30 pigs (31.1 ± 1.2 kg initial body weight) were used to determine the performance depression because of HS. Treatments were: thermo neutral pigs fed a 22% CP control diet (TN-C); HS pigs fed the control diet (HS-C); HS pigs fed a 14% CP, AA supplemented diet (HS-AA). HS pigs had lower ADG and Lys utilization efficiency, and consumed 20 and 25% less Lys and Thr, respectively, than the TN-C pigs (P < 0.05). In Exp. 2 (comparative slaughter), 25 pigs (33.6 ± 0.65 kg initial body weight) were used to evaluate the effect of extra dietary AA either as protein-bound or free AA on the performance and carcass traits of HS pigs. Treatments were: control wheat-SBM-free Lys, Thr and Met diet (CON); diet with 30% more CP than CON (HSxP); diet added with free AA to contain at least 25% more of each AA than the recommended level (HSxAA). Ambient temperature (AT) ranged from 27.7 to 37.7°C, and body temperature (39.9 to 41.2°C) followed a similar daily pattern as the AT did. There was no dietary treatment effect on daily feed and NE intake (P > 0.10), but the Lys, Thr, and Met intake was higher in pigs fed the HSxP or HSxAA diets than in pigs fed the CON diet (P < 0.05). The daily weight gain (ADG) was not affected (P > 0.10) but G:F tended to be higher and the Lys utilization efficiency (ADG, g/g Lys intake) tended to be lower in HSxP pigs than in CON pigs (P < 0.10). The HSxAA pigs had higher ADG (P < 0.05), and tended to have higher weight of hot carcass and leg muscle, and the weight gain of hot carcass and leg muscle than the CON pigs (P < 0.10). The weight and daily weight gain of loin muscle was higher in the HSxAA than in the HSxP pigs (P < 0.05). Kidney weight and serum urea in HSxP pigs were higher than in CON and HSxAA pigs, but spleen weight was higher in HSxAA pigs than in CON and HSxP pigs (P < 0.05). These results confirm that HS reduces the VFI, and show that increased levels of AA either as free or protein-bound do not additionally reduce the VFI of HS pigs. These also show that extra free AA supply rather than protein-bound AA better ameliorate the reduced growth performance of HS pigs.

Recombinant Adeno-Associated Virus-mediated rescue of function in a mouse model of Dopamine Transporter Deficiency Syndrome.

Dopamine Transporter Deficiency Syndrome (DTDS) is a rare autosomal recessive disorder caused by loss-of-function mutations in dopamine transporter (DAT) gene, leading to severe neurological disabilities in children and adults. DAT-Knockout (DAT-KO) mouse is currently the best animal model for this syndrome, displaying functional hyperdopaminergia and neurodegenerative phenotype leading to premature death in ~36% of the population. We used DAT-KO mouse as model for DTDS to explore the potential utility of a novel combinatorial adeno-associated viral (AAV) gene therapy by expressing DAT selectively in DA neurons and terminals, resulting in the rescue of aberrant striatal DA dynamics, reversal of characteristic phenotypic and behavioral abnormalities, and prevention of premature death. These data indicate the efficacy of a new combinatorial gene therapy aimed at rescuing DA function and related phenotype in a mouse model that best approximates DAT deficiency found in DTDS.

Diffusion MRI: literature review in salivary gland tumors.

Surgical resection is currently the best treatment for salivary gland tumors. A reliable magnetic resonance imaging mapping, encompassing tumor grade, location, and extension may assist safe and effective tumor resection and provide better information for patients regarding potential risks and morbidity after surgical intervention. However, direct examination of the tumor grade and extension using conventional morphological MRI remains difficult, often requiring contrast media injection and complex algorithms on perfusion imaging to estimate the degree of malignancy. In addition, contrast-enhanced MRI technique may be problematic due to the recently demonstrated gadolinium accumulation in the dentate nucleus of the cerebellum. Significant developments in magnetic resonance diffusion imaging, involving voxel-based quantitative analysis through the measurement of the apparent diffusion coefficient, have enhanced our knowledge on the different histopathological salivary tumor grades. Other diffusion imaging-derived techniques, including high-order tractography models, have recently demonstrated their usefulness in assessing the facial nerve location in parotid tumor context. All of these imaging techniques do not require contrast media injection. Our review starts by outlining the physical basis of diffusion imaging, before discussing findings from diagnostic studies testing its usefulness in assessing salivary glands tumors with diffusion MRI.

Dietary levels of protein and free amino acids affect pancreatic proteases activities, amino acids transporters expression and serum amino acid concentrations in starter pigs.

The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA-supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat-soybean meal diet were measured in two 14-d trials. The LPAA diet contained free L-Lys, L-Thr, DL-Met, L-Leu, L-Ile, L-Val, L-His, L-Trp and L-Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b0,+ AT in duodenum, B0 AT1 in ileum (p < 0.05), and tended to express more y+ LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b0,+ AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b0,+ AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.

3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates.

3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.