Observações de cetáceos por iatistas no Atlântico Norte : um estudo piloto.

In the summer of 1998, yachtsmen sailing from the Caribbean to the Azores were encouraged to take part in an Atlantic cetacean survey. The principle aim of this project was to evaluate the potential of using regular seafarers as sources of data on cetacean distribution. Identification guides and sighting forms were distributed and participants were asked to record any cetacean sightings as well as to conduct routine set-effort watches. A secondary aim of the project was to investigate reports of illegal whaling in the Atlantic. Data collected from the yachtsmen reveal a concentration of sightings along the mid- Atlantic ridge, perhaps corresponding to an increase in productivity in this area. No further reports of whaling activity were made. Although this work only involves a small data set, it illustrates how useful yachtsmen can be in assisting research in otherwise inaccessible regions. Future involvement of yachtsmen in cetacean surveying should be encouraged, as long-term data sets gathered in this way can be invaluable in revealing offshore trends

A novel phosphorylation-independent interaction between SMG6 and UPF1 is essential for human NMD

Eukaryotic mRNAs with premature translation-termination codons (PTCs) are recognized and eliminated by nonsense-mediated mRNA decay (NMD). NMD substrates can be degraded by different routes that all require phosphorylated UPF1 (P-UPF1) as a starting point. The endonuclease SMG6, which cleaves mRNA near the PTC, is one of the three known NMD factors thought to be recruited to nonsense mRNAs via an interaction with P-UPF1, leading to eventual mRNA degradation. By artificial tethering of SMG6 and mutants thereof to a reporter mRNA combined with knockdowns of various NMD factors, we demonstrate that besides its endonucleolytic activity, SMG6 also requires UPF1 and SMG1 to reduce reporter mRNA levels. Using in vivo and in vitro approaches, we further document that SMG6 and the unique stalk region of the UPF1 helicase domain, along with a contribution from the SQ domain, form a novel interaction and we also show that this region of the UPF1 helicase domain is critical for SMG6 function and NMD. Our results show that this interaction is required for NMD and for the capability of tethered SMG6 to degrade its bound RNA, suggesting that it contributes to the intricate regulation of UPF1 and SMG6 enzymatic activitie

Nonsense-mediated mRNA decay in human cells: mechanistic insights, functions beyond quality control and the double-life of NMD factors

Nonsense-mediated decay is well known by the lucid definition of being a RNA surveillance mechanism that ensures the speedy degradation of mRNAs containing premature translation termination codons. However, as we review here, NMD is far from being a simple quality control mechanism; it also regulates the stability of many wild-type transcripts. We summarise the abundance of research that has characterised each of the NMD factors and present a unified model for the recognition of NMD substrates. The contentious issue of how and where NMD occurs is also discussed, particularly with regard to P-bodies and SMG6-driven endonucleolytic degradation. In recent years, the discovery of additional functions played by several of the NMD factors has further complicated the picture. Therefore, we also review the reported roles of UPF1, SMG1 and SMG6 in other cellular processe

3D ultrasound reconstruction of sonographic callus:a novel imaging modality for early evaluation of fracture healing

AIMS: The aim of this study was to establish a reliable method for producing 3D reconstruction of sonographic callus. METHODS: A cohort of ten closed tibial shaft fractures managed with intramedullary nailing underwent ultrasound scanning at two, six, and 12 weeks post-surgery. Ultrasound capture was performed using infrared tracking technology to map each image to a 3D lattice. Using echo intensity, semi-automated mapping was performed to produce an anatomical 3D representation of the fracture site. Two reviewers independently performed 3D reconstructions and kappa coefficient was used to determine agreement. A further validation study was undertaken with ten reviewers to estimate the clinical application of this imaging technique using the intraclass correlation coefficient (ICC). RESULTS: Nine of the ten patients achieved union at six months. At six weeks, seven patients had bridging callus of ≥ one cortex on the 3D reconstruction and when present all achieved union. Compared to six-week radiographs, no bridging callus was present in any patient. Of the three patients lacking sonographic bridging callus, one went onto a nonunion (77.8% sensitive and 100% specific to predict union). At 12 weeks, nine patients had bridging callus at ≥ one cortex on 3D reconstruction (100%-sensitive and 100%-specific to predict union). Presence of sonographic bridging callus on 3D reconstruction demonstrated excellent reviewer agreement on ICC at 0.87 (95% confidence interval 0.74 to 0.96). CONCLUSION: 3D fracture reconstruction can be created using multiple ultrasound images in order to evaluate the presence of bridging callus. This imaging modality has the potential to enhance the usability and accuracy of identification of early fracture healing. Cite this article: Bone Joint Res 2021;10(12):759–766

Single Eye mRNA-Seq Reveals Normalisation of the Retinal Microglial Transcriptome Following Acute Inflammation

Background: Whether retinal microglia can maintain or restore immune homeostasis during and after inflammation is unclear. We performed single-eye mRNA-sequencing on microglia at different timepoints following a single inflammatory stimulus to characterise their transcriptome during and after resolution of endotoxin-induced uveitis (EIU). / Experimental Approach: Cx3cr1CreER:R26-tdTomato (C57BL/6) male heterozygotes were administered tamoxifen via different regimes at 4–5 weeks of age. Four weeks post-tamoxifen, mice were injected intravitreally with 10 ng lipopolysaccharide (endotoxin induced uveitis, EIU). Six-hundred retinal microglia were obtained by FACS from individual naïve retinas and at 4 h, 18 h, and 2 weeks following EIU induction. Samples were sequenced to a depth of up to 16.7 million reads using the SMART-Seq v4 Ultra Low Input RNA kit. The data was analysed using Partek software and Ingenuity Pathway Analysis. Genes were considered differentially-expressed (DEG) if the FDR step-up p-value was ≤0.05 and the fold-change was ≥±2. / Results: Flow cytometric analysis indicates that the Cx3cr1CreER:R26-tdTomato strain is both sensitive (>95% tagging) and specific (>95% specificity) for microglia when tamoxifen is administered topically to the eye for 3 days. During “early” activation, 613 DEGs were identified. In contrast, 537 DEGs were observed during peak cellular infiltrate and none at 2 weeks, compared to baseline controls (1,069 total unique DEGs). Key marker changes were validated by qPCR, flow cytometry, and fluorescence microscopy. C5AR1 was identified and validated as a robust marker of differentiating microglial subsets during an LPS response. / Conclusion: Using EIU to provide a single defined inflammatory stimulus, mRNA-Seq identified acute transcriptional changes in retinal microglia which returned to their original transcriptome after 2 weeks. Yolk-sac derived microglia are capable of restoring their homeostatic state after acute inflammation

Cytokine Response to Traditional and Cluster Sets in Resistance-trained Women

Resistance exercise that incorporates intra-set rest between repetition blocks (i.e., cluster sets [CS]) can produce a smaller metabolic stress and endocrine response than traditional sets (TS). PURPOSE: To examine the effect of CS on the acute cytokine response in resistance trained women. METHODS: 12 resistance-trained women (mean ± SE; 23.7 ± 1.1 years; 160.1 ± 1.5 cm; 62.5 ± 1.7 kg; 5 ± 1 years training) completed 3 sessions in the follicular phase. One-repetition maximum (1RM) back squat (BS) (98.7 ± 4.1 kg), and BS:body mass (1.6 ± 0.1) were determined in Session 1. For Session 2 (3 days post Session 1) and Session 3 (7 days post Session 2), subjects were randomly assigned to either 4 sets of 10 reps with 120 seconds (s) inter-set rest (TS) or 4 x (2 x 5 reps) with 30s intra-set rest and 90s inter-set rest (CS). All performed both protocols at 70% 1RM BS. Instructions were to perform every rep “as explosively as possible”. Blood was collected pre-exercise (PRE), immediately after sets 1, 2, 3, 4 (IP), and at 5 (+5), 15 (+15), 30 (+30), and 60 (+60) min post-exercise and analyzed for interleukin (IL)-1β, IL-2, IL-6, IL-8, IL 10, and IL-15. Data were analyzed using repeated measures ANOVAs (2 × 9). RESULTS: A significant main effect of time (p\u3c0.05) was found for IL-1β, IL-2, IL-8, IL-10, and IL-15. Concentration of IL-1β was smaller at +5 (3.9 ± 0.4 ng/mL), +15 (3.6 ± 0.4) +30 (3.5 ± 0.3), and +60 (3.7 ± 0.4) compared to IP (4.1 ± 0.4). IL-2 was greater after set 1 (10.8 ± 1.0 ng/mL), and set 2 (11.0 ± 1.2) compared to PRE (10.2 ± 1.0), and smaller at +30 (9.9 ± 1.0) compared to IP (11.0 ± 1.0). IL-8 was greater after set 1 (8.4 ± 0.6 ng/mL), set 2 (8.6 ± 0.7), and set 3 (8.5 ± 0.7) compared to PRE (8.0 ± 0.6). IL-10 was smaller at +30 (31.3 ± 7.4 ng/mL) compared to PRE (34.0 ± 7.4), and also smaller at +15 (32.6 ± 7.9) +30 (31.3 ± 7.4), and +60 (33.4 ± 8.6) compared to IP (38.0 ± 8.6). IL-15 was greater at IP (15.5 ± 4.0 ng/mL) compared to PRE (13.4 ± 3.5), and smaller at PRE (13.4 ± 3.5), +30 (11.9 ± 3.3), and +60 (11.6 ± 3.2) compared to IP (15.5 ± 4.0). No condition × time point effects were observed. CONCLUSION: Both TS and CS induced an acute cytokine response in resistance-trained women; incorporating intra-set rest (CS) did not appear to affect this cytokine response