Advances in Antisense Oligonucleotide Development for Target Identification, Validation, and as Novel Therapeutics

Antisense oligonucleotides (As-ODNs) are single stranded, synthetically prepared strands of deoxynucleotide sequences, usually 18–21 nucleotides in length, complementary to the mRNA sequence of the target gene. As-ODNs are able to selectively bind cognate mRNA sequences by sequence-specific hybridization. This results in cleavage or disablement of the mRNA and, thus, inhibits the expression of the target gene. The specificity of the As approach is based on the probability that, in the human genome, any sequence longer than a minimal number of nucleotides (nt), 13 for RNA and 17 for DNA, normally occurs only once. The potential applications of As-ODNs are numerous because mRNA is ubiquitous and is more accessible to manipulation than DNA. With the publication of the human genome sequence, it has become theoretically possible to inhibit mRNA of almost any gene by As-ODNs, in order to get a better understanding of gene function, investigate its role in disease pathology and to study novel therapeutic targets for the diseases caused by dysregulated gene expression. The conceptual simplicity, the availability of gene sequence information from the human genome, the inexpensive availability of synthetic oligonucleotides and the possibility of rational drug design makes As-ODNs powerful tools for target identification, validation and therapeutic intervention. In this review we discuss the latest developments in antisense oligonucleotide design, delivery, pharmacokinetics and potential side effects, as well as its uses in target identification and validation, and finally focus on the current developments of antisense oligonucleotides in therapeutic intervention in various diseases

Dynamic deformation of metastable austenitic stainless steels at the nanometric length scale

Cyclic indentation was used to evaluate the dynamic deformation on metastable steels, particularly in an austenitic stainless steel, AISI 301LN. In this work, cyclic nanoindentation experiments were carried out and the obtained loading-unloading (or P-h) curves were analyzed in order to get a deeper knowledge on the time-dependent behavior, as well as the main deformation mechanisms. It was found that the cyclic P-h curves present a softening effect due to several repeatable features (pop-in events, ratcheting effect, etc.) mainly related to dynamic deformation. Also, observation by transmission electron microscopy highlighted that dislocation pile-up is the main responsible of the secondary pop-ins produced after certain cycles.Peer ReviewedPostprint (author's final draft

Studies on mastitis in cattle caused by environmental organisms

SIGLEAvailable from British Library Document Supply Centre- DSC:DX87088 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Animal-based measurements of the severity of mastitis in dairy cows

A range of clinical parameters were studied to assess their usefulness as objective markers of the severity of clinical mastitis in dairy cows. Cows with moderate clinical mastitis had significantly higher rectal temperatures and heart and respiratory rates than cows with mild clinical mastitis or control cows. The difference in temperature between quarters did not vary significantly between the control cows and the cows with mastitis, but there was a larger difference between quarter temperatures in the cows with moderate mastitis than in the cows with mild mastitis or the control cows. The hock-to-hock distance in the control cows was significantly smaller than in the cows with mild to moderate mastitis, but there was no significant difference in the distance either between the cows with mild mastitis and those with moderate mastitis, or between the cows with mastitis in the front quarters and those with mastitis in the hind quarters. The mechanical threshold to pain of the cows with mild and moderate mastitis was significantly lower than that of the control cow

The usefulness of a single measurement of insulin-like growth factor-1 as a predictor of embryo yield and pregnancy rates in a bovine MOET program

The objective was to determine if a single measurement of plasma insulin-like growth factor-1 (IGF-1) could predict the number of viable embryos obtained from donors and the likelihood of pregnancy in recipients in multiple ovulation and embryo transfer (MOET) programs in cattle. The embryo yields from 101 embryo recoveries were examined in maiden Holstein heifers (n=75) and multiparous Holstein cows (lactating cows n=20, dry cows n=6). Donors were super stimulated with FSH and embryo recovery was done non-surgically 7 days after artificial insemination. Embryos were classified according to the IETS criteria. Pregnancy rates in 100 maiden Holstein heifer recipients were analysed. Recipients were on day 7+/-1 of the estrous cycle at transfer. Pregnancy diagnosis was carried out at day 30 (PD 30) and rechecked at day 60 (PD 60) after transfer. Blood samples from coccygeal vessels taken at the time of embryo recovery (donors) and transfer (recipients) were analysed for IGF-1, insulin, beta-hydroxybutyrate (beta-OHB), non-esterified fatty acids (NEFA), urea and cholesterol. There was a negative correlation between the number of viable embryos and insulin (r=-0.33, P=0.025) in donor heifers. In donor cows, the number of viable embryos was correlated with IGF-1 (r=0.43, P=0.028) and cholesterol (r=-0.43, P=0.027). In recipients, PD30 and PD 60 were not affected by any of the circulating parameters analysed. Insulin, IGF-1 and cholesterol only explained 8.9, 13.9 and 15.8% of the variation in the production of viable embryos, respectively. Several factors affect MOET programs and under the circumstances of the present study the usefulness of hormonal and metabolic profiles as predictors of the outcome of this biotechnology was limited

Mortality of dairy sheep during the peri-parturient period: Results of a field investigation in greece

Objectives of the study were (a) to investigate the incidence risk of peri-parturient mortality in dairy flocks in Greece, (b) to study when such mortality occurs in relation to lambing, (c) to identify clinical problems associated with the deaths, and (d) to evaluate potentially associated factors. The deaths of ewes during the peri-parturient period (10 days before to 7 days after lambing) were monitored in 60 flocks over two years. In the ewes that died, detailed post-mortems were performed. The incidence risk of peri-parturient deaths was 0.7%. The daily incidence rate of deaths before, at, and after lambing was 0.05, 0.04, and 0.03%, respectively. Clinical problems most frequently associated with peri-parturient deaths were pregnancy toxaemia, ruminal acidosis and post-partum genital problems. For the occurrence of peri-parturient deaths in flocks, the average lambing rate in the flocks was the only significant factor. Regarding deaths before or after lambing, the season during which lambing took place and the number of animals in the flock, respectively, were found to be significant. Most ewes (54.5%) that died at lambing (dystocia) were primigravidae. © 2021 by the authors. Licensee MDPI, Basel, Switzerland