Effect of dietary l-glutamine supplementation on the intestinal physiology and growth during Solea senegalensis larval development.

The maturation of the intestinal digestive and absorptive functions might limit the amount of absorbed nutrients to fulfil the high requirements of the fast-growing marine fish larva. Glutamine (Gln) has been described to improve intestinal epithelium functions, due to its involvement in energy metabolism and protein synthesis. The purpose of this study was to evaluate dietary 0.2% Gln supplementation on aspects of intestinal physiology, protein metabolism and growth-related genes expression in Senegalese sole larvae. Experiment was carried out between 12 and 33 days post hatching (DPH) and fish were divided into two experimental groups, one fed Artemia spp. (CTRL) and the other fed Artemia spp. supplemented with Gln (GLN). GLN diet had two times more Gln than the CTRL diet. Samples were collected at 15, 19, 26 and 33 DPH for biometry, histology, and digestive enzymes activity, and at 33 DPH for gene expression, protein metabolism and AA content determination. Growth was significantly higher for Senegalese sole fed GLN diet, supported by differences on protein metabolism and growth-related gene expression. Slight differences were observed between treatments regarding the intestinal physiology. Overall, GLN diet seems to be directed to enhance protein metabolism leading to higher larval growth.

Effect of Dietary Incorporation of Yellow Mealworm as a Partial Fishmeal Replacer on Growth, Metabolism, and Intestinal Histomorphology in Juvenile Meagre (Argyrosomus regius).

Efforts have been made to find alternatives to fish meal (FM), as the sustainability of aquaculture depends on it. Insect meal (IM) is a potential candidate to partially replace FM, being more sustainable and economically viable. In this experimental trial, three diets were tested with different yellow mealworm incorporation: a control diet with no IM, a diet with an inclusion of 10% IM (Ins10), and a diet with an incorporation of 20% IM (Ins20). The diets were tested on 10.5 g meagre for 47 days. The results showed that an IM inclusion higher than 10% affected both growth (2.6 vs. 2.2) and FCR (1.5 vs. 1.9) of meagre juveniles. However, this reduction in growth did not result from lower protein retention or changes in muscle fibre area or density. Little differences were observed in the activity of pancreatic and intestinal enzymes except for aminopeptidase total activity which was higher in the control and Ins10 compared to Ins20 (3847 vs. 3540 mU/mg protein), suggesting no limitations in protein synthesis. Also, the alkaline phosphatase intestinal maturation index was higher in the control group compared to the IM groups (437 vs. 296). On the contrary, several differences were also found in the proteolytic activity in the hepatic and muscle tissues of meagre juveniles fed the Ins10 diet. The inclusion of IM had no impact on intestine histomorphology but changes were detected in the enterocytes of fish from control and Ins10 which showed hypervacuolization and nucleus misplacement compared to the Ins20 treatment. Nevertheless, a higher percentage of Vibrionaceae was recorded for meagre fed on the Ins20 diet. Since no signs of inflammation were observed in the distal intestine, this suggests IM incorporation could have had an important impact on intestinal health due to its antimicrobial properties. This is supported by an increase in the haematocrit in the treatments where IM was added (20 to 25%). In conclusion, incorporations of IM at percentages up to 10% do not seem to have a negative impact on meagre performance at this age but can enhance the fish immune system and protection against intestinal inflammation.

Postprandial pattern of digestive enzymes and protein turnover in meagre (Argyrosomus regius) juveniles.

After a meal, a sequence of physiological changes occurs in animals in response to digestion, absorption and assimilation of the ingested nutrients. These processes are very important for the aquaculture sector since they will define the efficiency by which food is converted into growth, thus contributing to reduce the production costs and also undigested food in the effluents. Here we investigated the activity of digestive enzymes in the midgut and the protein degradation systems following a single meal to define postprandial patterns of action in hepatic and muscle tissues of meagre juveniles. Fish were fed with a single meal followed by a period of 24 h without feed. The activity of several digestive enzymes (α-amylase, trypsin, aminopeptidase, alkaline and acid phosphatases) plus the analysis of key players of the ubiquitin-proteasome (UPS) and autophagy-lysosomal (ALS) systems were examined just before feeding (0 h, basal point) and 2, 4, 6, 8 and 24 h after food ingestion. Digestion was activated around 4 h after food ingestion and nutrients available for protein degradation 2 h later. This work provided information about the short-term physiological effects induced by a single meal to support scientists' decision when planning a specific study involving digestion and protein degradation, and also to fish farmers on how to better manage feeding protocols when producing A. regius juveniles. Our results suggested that meagre juveniles, under the experimental conditions used, can be fed every 4 h, time when digestion starts, however further studies should be addressed to find the optimal feeding regime for this juvenile species.

Outcome measures and motion capture systems for assessing lower limb orthosis-based interventions after stroke: a systematic review.

PURPOSE: To review and categorize, according to the International Classification of Functioning, the outcome measures, and motion capture systems for studying the evidence-based practice of orthotic-based interventions in post-stroke gait rehabilitation. METHODS: An electronic literature search was conducted up to February 2018 in Web of Science, Scopus, MEDLINE and Physiotherapy Evidence Database. Randomized trials measuring activity, impairment or participation outcome measures for studying the evidence-based practice of orthoses in gait rehabilitation after an acute or chronic stroke were identified. The studies were assessed through the Cochrane risk-of-bias tool by three authors. Information about stroke's stage, assessment protocol (goal, timing and motion capture system), orthosis configuration and outcome measures were extracted. RESULTS: Eighteen randomized trials, including 387 post-stroke adults, mostly in the chronic stage, were selected. They assessed 39 outcomes, mainly activity outcome measures such as spatiotemporal (72.2%), kinematic (44.4%) and functional (33.3%) outcomes. Gait speed was the primary outcome in most studies. Participation (22.2%) and impairment (16.7%) outcome measures were less explored. Mostly, non-portable motion capture systems were employed opposing the freely-use of the wearable orthosis. The detection bias risk and the shortage of baseline and follow-up outcome measures affected the studies' assessment quality. CONCLUSIONS: Studies showed heterogeneity in selecting outcomes and timings for assessment. There is evidence for assessing the evidence of orthosis-based gait rehabilitation after stroke through activity outcome measures, primarily the gait speed, recorded by non-wearable motion capture systems. A unified methodology considering wearable sensors for tracking baseline and follow-up measures is needed.Implications for rehabilitationThere is evidence on use activity outcome measures to assess the meaningful evidence-based practice of orthosis-based gait rehabilitation in post-stroke.Gait speed was the primary outcome measure most reported.There is limited evidence on use impairment and participation outcome measures to measure meaningful changes due to orthotic-based rehabilitation.Outcome measures were mainly recorded by non-wearable motion capture systems.

Ectopic expression of CITED2 prior to reprogramming, promotes and homogenises the conversion of somatic cells into induced pluripotent stem cells.

Cited2 plays crucial roles in mouse embryonic stem cells self-renewal, the initiation of the somatic reprogramming process into induced pluripotent stem cells (iPSC) and the suppression of cell senescence. Here, we investigated the potential of CITED2 expression in combination with the Oct4, Sox2, Klf4 and c-Myc factors for reprogramming of primary mouse embryonic fibroblasts (MEF) at passage 2 and 4. The ectopic CITED2 expression in primary MEF prior to the onset of the reprogramming process, generated iPSC with less variability in the expression of endogenous pluripotency-related genes. In contrast, part of the MEF reprogrammed without ectopic expression of CITED2 at passage 4 originated partially reprogrammed iPSC or pre-iPSC. However, the overexpression of CITED2 in the pre-iPSC was insufficient to complete the reprogramming process into iPSC. These results indicated that ectopic CITED2 expression at the onset of the reprogramming process in combination with the reprogramming factors promotes a complete and homogeneous conversion of somatic cells into iPSC.

CITED2 Cooperates with ISL1 and Promotes Cardiac Differentiation of Mouse Embryonic Stem Cells.

The transcriptional regulator CITED2 is essential for heart development. Here, we investigated the role of CITED2 in the specification of cardiac cell fate from mouse embryonic stem cells (ESC). The overexpression of CITED2 in undifferentiated ESC was sufficient to promote cardiac cell emergence upon differentiation. Conversely, the depletion of Cited2 at the onset of differentiation resulted in a decline of ESC ability to generate cardiac cells. Moreover, loss of Cited2 expression impairs the expression of early mesoderm markers and cardiogenic transcription factors (Isl1, Gata4, Tbx5). The cardiogenic defects in Cited2-depleted cells were rescued by treatment with recombinant CITED2 protein. We showed that Cited2 expression is enriched in cardiac progenitors either derived from ESC or mouse embryonic hearts. Finally, we demonstrated that CITED2 and ISL1 proteins interact physically and cooperate to promote ESC differentiation toward cardiomyocytes. Collectively, our results show that Cited2 plays a pivotal role in cardiac commitment of ESC.

FBXL5 modulates HIF-1α transcriptional activity by degradation of CITED2.

CITED2 is a ubiquitously expressed nuclear protein exhibiting a high affinity for the cysteine-histidine-rich domain 1 (CH1) of the transcriptional co-activators CBP/p300. CITED2 is particularly efficient in the inhibition of the hypoxia-inducible factor-1α (HIF-1α) dependent transcription by competing with it for the interaction with the CH1 domain. Here we report a direct and specific interaction between CITED2 and the F-box and leucine rich repeat protein 5 (FBXL5), a substrate adaptor protein which is part of E3 ubiquitin ligase complexes mediating protein degradation by the proteasome. We demonstrated that depletion of FBXL5 by RNA interference led to an increase of CITED2 protein levels. Conversely, overexpression of FBXL5 caused the decrease of CITED2 protein levels in a proteasome-dependent manner, and impaired the interaction between CITED2 and the CH1 domain of p300 in living cells. In undifferentiated mouse embryonic stem cells, the overexpression of FBXL5 also reduced Cited2 protein levels. Finally, we evidenced that FBXL5 overexpression and the consequent degradation of CITED2 enabled the transcriptional activity of the N-terminal transactivation domain of HIF-1α. Collectively, our results highlighted a novel molecular interaction between CITED2 and FBXL5, which might regulate the steady state CITED2 protein levels and contribute to the modulation of gene expression by HIF-1α.

Autonomic dysreflexia and posterior reversible encephalopathy syndrome.

Autonomic dysreflexia is a syndrome of massive imbalanced reflex sympathetic discharge in patients who had a spinal cord injury above the splanchnic sympathetic outflow resulting in a sudden increase in blood pressure. Posterior reversible encephalopathy syndrome (PRES) refers to a clinicoradiologic entity characterized by headache, consciousness impairment, visual disturbances, seizures, and posterior transient changes on neuroimaging (cerebral vasogenic edema). Hypertension is a common cause of PRES. The authors describe two case reports of patients with tetraplegia who developed PRES after an autonomic dysreflexia episode. One of them had recurrence of PRES in a similar clinical context. The authors discuss further aspects of PRES and its recurrence, which seems to be unusual particularly after autonomic dysreflexia.