Oligonucleotide-Palladacycle Conjugates as Splice-Correcting Agents

2'-O-Methylribo phosphorothioate oligonucleotides incorporating cyclopalladated benzylamine conjugate groups at their 5'-termini have been prepared and their ability to hybridize with a designated target sequence was assessed by conventional UV melting experiments. The oligonucleotides were further examined in splice-switching experiments in human cervical cancer (HeLa Luc/705), human liver (HuH7_705), and human osteosarcoma (U-2 OS_705) reporter cell lines. Melting temperatures of duplexes formed by the modified oligonucleotides were approximately 5 degrees C lower than melting temperatures of the respective unmodified duplexes. The cyclopalladated oligonucleotides functioned as splice-correcting agents in the HeLa Luc/705 cell line somewhat more efficiently than their unmodified counterparts. Furthermore, the introduction of this chemical modification did not induce toxicity in cells. These results demonstrate the feasibility of using covalently metalated oligonucleotides as therapeutic agents

Oligonucleotides Targeting DNA Repeats Downregulate Huntingtin Gene Expression in Huntington's Patient-Derived Neural Model System

Huntington's disease (HD) is one of the most common, dominantly inherited neurodegenerative disorders. It affects the striatum, cerebral cortex, and other subcortical structures leading to involuntary movement abnormalities, emotional disturbances, and cognitive impairments. HD is caused by a CAG•CTG trinucleotide-repeat expansion in exon 1 of the huntingtin (HTT) gene leading to the formation of mutant HTT (mtHTT) protein aggregates. Besides the toxicity of the mutated protein, there is also evidence that mtHTT transcripts contribute to the disease. Thus, the reduction of both mutated mRNA and protein would be most beneficial as a treatment. Previously, we designed a novel anti-gene oligonucleotide (AGO)-based strategy directly targeting the HTT trinucleotide-repeats in DNA and reported downregulation of mRNA and protein in HD patient fibroblasts. In this study, we differentiate HD patient-derived induced pluripotent stem cells to investigate the efficacy of the AGO, a DNA/Locked Nucleic Acid mixmer with phosphorothioate backbone, to modulate HTT transcription during neural in vitro development. For the first time, we demonstrate downregulation of HTT mRNA following both naked and magnetofected delivery into neural stem cells (NSCs) and show that neither emergence of neural rosette structures nor self-renewal of NSCs is compromised. Furthermore, the inhibition potency of both HTT mRNA and protein without off-target effects is confirmed in neurons. These results further validate an anti-gene approach for the treatment of HD

Effect of long-term betaine supplementation on chemical and physical characteristics of three muscles from the Alentejano pig

BACKGROUND: The use of betaine as a dietary supplement to reduce fat deposition and increase lean muscle mass in pigs is increasing. However, there is no available information on its effect on the physical and chemical (i.e. fatty acid (FA) profile) characteristics of pork. RESULTS: The effects of long-term betaine intake (1 g betaine kg−1 diet for 20 weeks) on the chemical and physical characteristics of Musculus Longissimus dorsi, M. biceps femoris and M. semimembranosus from the Alentejano pig were investigated. The contents of total protein, intramuscular lipids (neutral and polar), myoglobin and total collagen as well as the water-holding capacity, physical colour characteristics and FA profile of the three muscles were determined. With the exception of a significant increase in the intramuscular lipid content of M. longissimus dorsi and M. biceps femoris, betaine supplementation did not influence muscle chemical and colour characteristics or body fat deposition. CONCLUSION: These data suggest that long-term betaine supplementation selectively increases intramuscular lipid deposition (a trait related to meat quality) while not affecting other chemical (including the atheroprotective FA profile) and physical characteristics of the Alentejano pig muscles studied

ANKRD54 preferentially selects Bruton's Tyrosine Kinase (BTK) from a Human Src-Homology 3 (SH3) domain library

Bruton's Tyrosine Kinase (BTK) is a cytoplasmic protein tyrosine kinase with a fundamental role in B-lymphocyte development and activation. The nucleocytoplasmic shuttling of BTK is specifically modulated by the Ankyrin Repeat Domain 54 (ANKRD54) protein and the interaction is known to be exclusively SH3-dependent. To identify the spectrum of the ANKRD54 SH3-interactome, we applied phage-display screening of a library containing all the 296 human SH3 domains. The BTK-SH3 domain was the prime interactor. Quantitative western blotting analysis demonstrated the accuracy of the screening procedure. Revealing the spectrum and specificity of ANKRD54-interactome is a critical step toward functional analysis in cells and tissues.Peer reviewe

Low-intensity blue-enriched white light (750 lux) and standard bright light (10 000 lux) are equally effective in treating SAD. A randomized controlled study

<p>Abstract</p> <p>Background</p> <p>Photoreceptor cells containing melanopsin play a role in the phase-shifting effects of short-wavelength light. In a previous study, we compared the standard light treatment (SLT) of SAD with treatment using short-wavelength blue-enriched white light (BLT). Both treatments used the same illuminance (10 000 lux) and were equally highly effective. It is still possible, however, that neither the newly-discovered photoreceptor cells, nor the biological clock play a major role in the therapeutic effects of light on SAD. Alternatively, these effects may at least be partly mediated by these receptor cells, which may have become saturated as a result of the high illuminances used in the therapy. This randomized controlled study compares the effects of low-intensity BLT to those of high-intensity SLT.</p> <p>Method</p> <p>In a 22-day design, 22 patients suffering from a major depression with a seasonal pattern (SAD) were given light treatment (10 000 lux) for two weeks on workdays. Subjects were randomly assigned to either of the two conditions, with gender and age evenly distributed over the groups. Light treatment either consisted of 30 minutes SLT (5000°K) with the EnergyLight^®(Philips, Consumer Lifestyle) with a vertical illuminance of 10 000 lux at eye position or BLT (17 000°K) with a vertical illuminance of 750 lux using a prototype of the EnergyLight^®which emitted a higher proportion of short-wavelengths. All participants completed questionnaires concerning mood, activation and sleep quality on a daily basis. Mood and energy levels were also assessed on a weekly basis by means of the SIGH-SAD and other assessment tools.</p> <p>Results</p> <p>On day 22, SIGH-SAD ratings were significantly lower than on day 1 (SLT 65.2% and BLT 76.4%). On the basis of all assessments no statistically significant differences were found between the two conditions.</p> <p>Conclusion</p> <p>With sample size being small, conclusions can only be preliminary. Both treatment conditions were found to be highly effective. The therapeutic effects of low-intensity blue-enriched light were comparable to those of the standard light treatment. Saturation effects may play a role, even with a light intensity of 750 lux. The therapeutic effects of blue-enriched white light in the treatment of SAD at illuminances as low as 750 lux help bring light treatment for SAD within reach of standard workplace and educational lighting systems.</p

Effect of different levels of synthetic astaxanthin on growth, skin color and lipid metabolism of commercial sized red porgy (Pagrus pagrus)

A study was undertaken to evaluate the effects of supplementing commercial feeds with Car ophyll® Pink, a gelatin-encapsuled astaxanthin, on growth parameters, lipid composition, pigment concentration and skin color of commercial size red porgy. Three isonitrogenous diets were formulated containing different levels of astaxantin: Ctrl diet (no pigment added), A50 diet (with 50 mg/kg of astaxanthin) and A80 diet (with 80 mg/ kg of astaxanthin). Four groups of fish (386.29 ± 29.50 g initial weight) were grown in triplicate tanks (10 fish per tank). Ctrl, A50 and A80 groups were fed the respective diets to apparent satiation for 180 days, while a fourth group was fed A50 diet for 90 days followed by A80 diet for another 90 days (A50A80). At the end of trial muscle samples were collected for composition analysis, liver and mesenteric fat for lipid composition determination and skin samples for carotenoids quantification. Dietary astaxanthin supplementation had no effect on growth performance and hepatosomatic index. However, A50A80 group displayed a lower muscle protein content and higher fat content when compared to the other groups. There was no clear effect of dietary astaxanthin supplementation on the liver’s and mesenteric fatty acid profile. Although there was no significant effect on skin total carot enoids, the astaxanthin content tended to increase in fish fed astaxanthin supplemented diets, which was reflected on redness values. Accordingly, dietary astaxanthin supplementation (at 50 or 80 mg/kg for 6 months or at 50 mg/kg for 3 months followed by 80 mg/kg for 3 months) influenced positively skin hue and chroma of red porgy on both pectoral and caudal areas, improving the skin color in commercial sized red porgy and achieving hue and chroma values close to those previously reported for wild individuals. The results reported here provide evidence of the applicability of Carophyll® Pink to improve skin color of large size commercial red porgy, but further studies are needed to optimize this carotenoid supplementation in production farming.info:eu-repo/semantics/publishedVersio

GW190412: observation of a binary-black-hole coalescence with asymmetric masses

We report the observation of gravitational waves from a binary-black-hole coalescence during the first two weeks of LIGO’s and Virgo’s third observing run. The signal was recorded on April 12, 2019 at 05∶30∶44 UTC with a network signal-to-noise ratio of 19. The binary is different from observations during the first two observing runs most notably due to its asymmetric masses: a ∼30 M⊙ black hole merged with a ∼8 M⊙ black hole companion. The more massive black hole rotated with a dimensionless spin magnitude between 0.22 and 0.60 (90% probability). Asymmetric systems are predicted to emit gravitational waves with stronger contributions from higher multipoles, and indeed we find strong evidence for gravitational radiation beyond the leading quadrupolar order in the observed signal. A suite of tests performed on GW190412 indicates consistency with Einstein’s general theory of relativity. While the mass ratio of this system differs from all previous detections, we show that it is consistent with the population model of stellar binary black holes inferred from the first two observing runs