Effects of Pair versus Individual Housing on Romanian Simmental x Limousine Crossbreed Calves Behaviour

The aim of the study was to assess the effects of housing method on Romanian Simmental x Limousine (RS x L) crossbred calves behaviour. To investigate these effects 12 RS x L calves were separated from damns within the first hour after birth and were housed either individually (n=6) or in pair (n=3 pairs) on straw bedded pens. The inactive standing, habitat exploring and playing behaviours were recorded on days 15, 30, 45 and 60 after birth. Individually housed calves spent more time (P≤0.016) standing inactive (means ±SEM) 5.16±0.47 bouts / day compared to pair housed calves (3.12±0.57 bouts / day). In gregarious animals, presence of a partner reduces stress and fear due to the social isolation. The pair housed calves spent significantly less time (P≤0.039) to explore the habitat (3.5±0.56 bouts / day) compared to individually housed calves, which resulted in higher frequency of exploration bouts (5.83±0.79 bouts / day). Also, paired calves spent more time (P≤0.001) in social contact, playing with the partner (4.16±0.7 bouts / day) compared with individually housed calves (0.16±0.06 bouts / day). These results indicate that housing calves in pairs generated benefits for calves such as better welfare condition, social opportunities and expression of desired behaviours patterns

Mammalian Oocyte Cryopreservation - Review

Although oocyte cryopreservation represents one of the main objectives of the reproductive medicine and cryobiology in the last years, until now, regardless of the studied specie, there is no freezing protocol that will assure satisfactory survival rates after thawing. Oocyte cryopreservation is now one of the most problematic issues in cryobiology aria, especially because they are extremely sensitive to low temperatures, and the maintaining of a normal development potential after thawing is very difficult. For animals cryopreservation is offering the possibility of preserving the genetically valuable genotypes or endangered species, with reducing the costs associated with conventional breeding (shelter, food, bed, drugs, human resources, etc.). The main factors affecting the viability of the oocyte are cryoprotectors, freezing method, disruption of meiotic spindle, zona pellucida hardening and oocyte maturation stage. The present paper is aiming to review the historic of oocyte cryopreservation, the main factor involved in oocyte viability and current trends in gamete preservation

Comparative study of the differentiation potential of rat bone marrow mesenchymal stem cells and rat muscle-derived stem cells

We present a comparative study of the plasticity of rat bone marrow mesenchymal stem cells (MSCs) and rat muscle-derived stem cells (MDSCs). The study was performed on two cell populations that were isolated by aspiration from the femur bone marrow and gastrocnemius muscle biopsy of 6-week-old albino rats. Both cell populations were exposed to identical stimulation conditions. The cells were capable of undergoing osteogenic, chondrogenic, adipogenic and epithelial differentiation, as shown by histochemistry and immunostaining techniques. The MDSC population showed behavior and characteristics similar to the bone marrow MSC population; however, the osteogenic and adipogenic potential was more reduced compared to MSCs. Our results indicate a positive expression of E cadherin and Cytokeratin 10 after 28 days under epithelial stimulation, suggesting a potential use for gastrocnemius muscle MDSCs as a promising source for regenerative therapies, including re-epithelialization and skin regeneration

Effect of ß-lactoglobulin Locus Polymorphism on Milk Related Traits in Romanian Spotted

The main aim of the study was to assess the influence of ß-lactoglobulin (LGB) genotype on milk related traits in Romanian Spotted (R.S.) breed. Altogether 254 cattle were genotyped for in order to establish the share of A and B allele in LGB locus using PCR-RFLP assay. The most prevalent was A allele (0.662) compared to B allele (0.338). Comparable frequencies (P>0.084) were recorded for AA (0.434) and AB (0.455) genotypes. The BB heterozygous recorded a lower frequency (0.111) compared with others (P≤0.001). For the LGB polymorphism, no significant differences (P>0.05) were observed according to milk production. The higher milk production was associated to AB (6094.31±103.22 kg) compared to AA (5912.22±91.7 kg, P>0.53) and BB (5977.7±81.12 kg, P>0.71) genotypes. The higher fat percentage (4.26±0.02%) was recorded for BB genotype, compared to AA genotype (4.19±0.02%, (P≤0.019). A significantly increased protein percentage was associated with AB genotype (3.43±0.03%) compared with AA (3.28±0.02%, P≤0.027). No significant difference (P>0.66) was recorded compared to BB genotype (3.42±0.01%) related to this trait. The results obtained encourage including marker assisted-selection and use the genotyped sires for genes with economic values in the future breeding scheme of Romanian Spotted breed

PLURIPOTENT STEM CELLS FROM THE ADULT MOUSE UTRICLE

Researchers discovered that cochlear epithelia in mice especially the vestibular one, contains stem cells that have the capacity to differentiate in sensorial auditory hair cell progenitors specific to the organ. They are reduced in number as the animal progresses in age. This process leads to a loss in the regenerative and proliferative potential of sensorial inner ear epithelia secondary to different injuries. Isolation, cultivation and than in vitro differentiation of vestibular stem cells could become a regenerative implant for acquired hearing loss. These were the motives that determined us to try to isolate, cultivate and finally differentiate vestibular stem cells from vestibular epithelia.Utricles from 7 days old mice NMRI were harvested, the otolites were removed, the utricles were trypsinized in order to isolate cells. Obtained cells were cultivated at 37ºC and 5% CO2 in DMEM with F12 Nutrient mixture, B27, N2 supplement. Pluripotency of obtained spheres was established with the help of stem cell markers Nanog and Oct-4. For identification of progenitor cells we used the marker, which reveals the gene which encodes the protein nestin. In all experiments we obtained floating colonies called spheres, formed by mitotic multiplying. For testing the pluripotency of spheres we used Nanog and Oct-4, two transcription factors that are expressed at high levels in stem cells and that we found to be expressed in our spheres. The presence of nestin mRNA in cells composing the spheres showed that these progressed to a progenitor cell stage.We concluded that utricular epithelia in 7 days old mice contains sufficient stem cells that can be cultivated and that can be later differentiated

TargetPlex FFPE-Direct DNA Library Preparation Kit for SiRe NGS panel: an international performance evaluation study

Next generation sequencing (NGS) represents a key diagnostic tool to identify clinically relevant gene alterations for treatment-decision making in cancer care. However, the complex manual workflow required for NGS has limited its implementation in routine clinical practice. In this worldwide study, we validated the clinical performance of the TargetPlex FFPE-Direct DNA Library Preparation Kit for NGS analysis. Impressively, this new assay obviates the need for separate, labour intensive and time-consuming pre-analytical steps of DNA extraction, purification and isolation from formalin-fixed paraffin embedded (FFPE) specimens in the NGS workflow

TargetPlex FFPE-Direct DNA Library Preparation Kit for SiRe NGS panel: an international performance evaluation study

Aim Next generation sequencing (NGS) represents a key diagnostic tool to identify clinically relevant gene alterations for treatment-decision making in cancer care. However, the complex manual workflow required for NGS has limited its implementation in routine clinical practice. In this worldwide study, we validated the clinical performance of the TargetPlex FFPE-Direct DNA Library Preparation Kit for NGS analysis. Impressively, this new assay obviates the need for separate, labour intensive and time-consuming pre-analytical steps of DNA extraction, purification and isolation from formalin-fixed paraffin embedded (FFPE) specimens in the NGS workflow. Methods The TargetPlex FFPE-Direct DNA Library Preparation Kit, which enables NGS analysis directly from FFPE, was specifically developed for this study by TargetPlex Genomics Pleasanton, California. Eleven institutions agreed to take part in the study coordinated by the Molecular Cytopathology Meeting Group (University of Naples Federico II, Naples, Italy). All participating institutions received a specific Library Preparation Kit to test eight FFPE samples previously assessed with standard protocols. The analytical parameters and mutations detected in each sample were then compared with those previously obtained with standard protocols. Results Overall, 92.8% of the samples were successfully analysed with the TargetPlex FFPE-Direct DNA Library Preparation Kit on Thermo Fisher Scientific and Illumina platforms. Altogether, in comparison with the standard workflow, the TargetPlex FFPE-Direct DNA Library Preparation Kit was able to detect 90.5% of the variants. Conclusion The TargetPlex FFPE-Direct DNA Library Preparation Kit combined with the SiRe panel constitutes a convenient, practical and robust cost-saving solution for FFPE NGS analysis in routine practice