Cell therapy of alpha-sarcoglycan null dystrophic mice through intra-arterial delivery of mesoangioblasts

Preclinical or clinical trials for muscular dystrophies have met with modest success, mainly because of inefficient delivery of viral vectors or donor cells to dystrophic muscles. We report here that intra-arterial delivery of wild-type mesoangioblasts, a class of vessel-associated stem cells, corrects morphologically and functionally the dystrophic phenotype of virtually all downstream muscles in adult immunocompetent alpha-sarcoglycan (alpha-SG) null mice, a model organism for limb-girdle muscular dystrophy. When mesoangioblasts isolated from juvenile dystrophic mice and transduced with a lentiviral vector expressing alpha-SG were injected into the femoral artery of dystrophic mice, they reconstituted skeletal muscle in a manner similar to that seen in wild-type cells. The success of this protocol was mainly due to widespread distribution of donor stem cells through the capillary network, a distinct advantage of this strategy over previous approaches.     

Effect of antioxidant protection of must on volatile compounds and aroma shelf life of Falanghina (Vitis vinifera L.) wine

Two vinification methods involving different degrees of antioxidant protection of Falanghina must during prefermentative steps, and referred as HAMP (high antioxidant must protection) and LAMP (low antioxidant must protection), were compared in terms of fermentation performances of four different yeast strains, composition of the volatile fraction of wines at the end of alcoholic fermentation, and shelf life of wines during storage. The use of HAMP technology resulted in wines with lower volatile acidity and higher concentrations of medium-chain fatty acid ethyl esters, acetates, and volatile fatty acids. For two of the four strains a lower concentration of isoamyl alcohol was also observed. HAMP wines also revealed increased shelf life because of the higher concentration of odor active esters at the end of storage and better preservation of varietal aromas.     

Microwave-assisted Acid decomposition of animal- and plant-derived samples for element analysis

A single microwave-assisted acid-decomposition procedure is proposed for sample preparation of plant- and bovine-derived materials prior to multielement determination by inductively coupled plasma-optical emission spectrometry (ICP-OES). The procedure involved sample grinding in a cryogenic mill to reduce particle sizes and increase the surface area for acid attack, followed by decomposition under high pressure and temperature. A single heating program was employed and efficiently destroyed most of the organic matrix by using small volumes of nitric acid and hydrogen peroxide. Accuracy of the elemental analysis was indicated by the good agreement between certified and found values of elements in plant and animal samples at 95% confidence level. The sample preparation procedure was fast and reproducible, and proved to be efficient for elemental analysis in agricultural and food-related samples by a sensitive multielement technique such as ICP-OES.     

New self-incompatibility alleles in apricot (Prunus armeniaca L.) revealed by stylar ribonuclease assay and S-PCR analysis

Apricot (Prunus armeniaca L.) shows gametophytic self-incompatibility controlled by a single locus with several allelic variants. An allele for self-compatibility (S_C) and seven alleles for self-incompatibility (S₁–S₇) were described previously. Our experiments were carried out to ascertain whether the number of allelic variants of apricot S-locus was indeed so small. Twenty-seven apricot accessions were analysed for stylar ribonucleases by non-equilibrium pH gradient electrofocusing (NEpHGE) to determine their S-genotype. To validate the results of electrofocusing, the applicability of the S-gene-specific consensus PCR primers designed from sweet cherry sequences was tested. NEpHGE revealed 12 bands associated with distinct S-alleles in newly genotyped cultivars. Cherry consensus primers amplified 11 alleles out from 16 ones, which indicated that these primers could also recognize most of the S-RNase sequences in apricot, and provided an efficient tool to confirm or reject NEpHGE results. By combining the protein and DNA-based methods, complete or partial S-genotyping was achieved for 23 apricot accessions and nine putatively new alleles (provisionally labelled S₈–S₁₆) were found. Their identity needs to be confirmed by pollination tests or S-allele sequencing. This study provides evidence that similarly to other Prunus species, the S-locus of apricot is more variable than previously believed.     

In vitro culture of female date palm (Phoenix dactylifera L.) tissues

Summary Roots, leaf petioles, shoot tips and immature fruits of female date palm formed the source material for in vitro cultures. Isolated embryos excised from mature seeds were also cultured. Of all the tissues, some of the leaf petioles and fruit mesocarp gave rise to calluses in the media of Staritsky (1970) and Eeuwens (1976). Isolated embryos germinated to produce seedlings in MS (Murashige & Skoog, 1962) medium. Calluses were established from seedling sections but the growth conditions propitious for explants of seedlings were not suitable for female date palm tissues. The main problem in establishing long term tissue cultures has been the browning of the tissues and media. This problem was, at least partially, overcome by adding activated charcoal to the liquid media and cysteine to the solid madia. So far, shoot regeneration in the calluses could not be induced.     

Spectral Doppler ultrasound in selecting an equine embryo receiver

The aim of this study was to evaluate whether the RI and PI values would help in choosing the best embryo recipient, and observe whether CL vascularization would influence P4 production. During the breeding season 2018/2019, the study was conducted using 35 mares, which is used for reference to collect data for the project on the day of embryo transfer. The utilized mares were divided into five groups followed by the day after ovulation, with D0 being the day of ovulation. Therefore, the five groups are as follows: D4—mares that were on the 4th post-ovulation day; D5—mares that were on the 5th post-ovulation day; and doing so successively for D6, D7 and D8. On the day of embryo transfer, the CL of the mares that selected as recipients was evaluated by B-mode and power flow mode ultrasonography and the right and left dorsal branches of the uterine arteries by spectral Doppler ultrasonography. Blood samples were taken on the day of the embryo transfer for a dosage of P4 concentration by radioimmunoassay. No statistical difference was found between the variables when the mares were separated into pregnant and non-pregnant mares, or when they were separated by age groups. When the groups of mares were compared by the day of embryo transfer, the statistical difference was found between the groups D5 × D6 (p = .0053) and D6 × D8 (p = .0036) in RI variable. In PI variable, the statistical difference was found between the groups D4 × D8 (p = .049), D5 × D6 (p = .0446) and D6 × D8 (p = .0024). We conclude that the mares with RI measurement of uterine arteries near 1.0 are correlated to mares with high CL vascularization and elevated P4 concentration.     

Ovum pick-up and in vitro maturation in spotted paca (Cuniculus paca—Linnaeus, 1766)

We tested FSHp, eCG and FSHp + eCG to establish ovum pick-up (OPU) and in vitro maturation method in spotted paca. Eight healthy adult females were subjected to each of four treatments to stimulate ovarian follicular growth. All females were subjected to a hormonal protocol using a single dose of 45 mg of injectable progesterone and single intramuscular injection of 0.075 mg d-cloprostenol on day 6. Ovarian stimulation was carried out as follows: in Group TFE (FSHp and eCG), animals were treated with a single dose of 80 mg of FSHp and 200 IU of eCG intramuscularly on day 6 after the application of progesterone; in Group TF (FSHp), they were treated with a single dose of 80 mg of FSHp intramuscularly on day 6 after application of progesterone; in Group treatment eCG, they were treated with 200 IU of eCG intramuscularly on day 6 after application of progesterone; and in Group TC (saline solution), 1 ml of saline solution was administered to control does. The OPU was performed between 22 and 26 hr after gonadotropin treatments. All recovered oocytes were placed into maturation media and incubated for 24 hr. There were no differences among the mean number of observed follicles, aspirated follicles and oocytes recovered per treatment. Oocyte maturation rates did not differ among groups, except, TF and treatment eCG oocytes had greater maturation rates than TC oocytes. In this study, gonadotropin administration failed to superovulate treated does and increase oocyte retrieval efficiency. Despite the feasibility of the procedure, further studies are needed to develop and refine hormonal protocols for oocyte recovery and in vitro maturation in this species.     

Bryde's whales in the North-East Atlantic: New insights on site fidelity and connectivity between oceanic archipelagos

1. The conservation of marine megafauna presents numerous difficulties owing to their high mobility over difficult-to-access oceanic areas that impairs the collection of basic, but essential, biological information.
2. The Bryde's whale (Balaenoptera edeni) is one of the most elusive species of baleen whales, and although it is known to be a seasonal visitor to several archipelagos in Macaronesia (the Azores, Madeira, and Canaries), there are no studies regarding its occurrence or geographical connectivity in this area of the Atlantic.
3. A 14-year photographic database was used to determine short-term (intra-seasonal) and long-term (inter-annual) Bryde's whale site fidelity and to estimate individual residency times in Madeira, whereas photographic catalogues from Madeira and the Canaries were compared in order to assess large-scale movements (i.e. on the scale of hundreds of kilometres).
4. In Madeira, 59 individuals were identified, 27 (45.8%) of which were recaptured. Of these, 10 individuals (37.0%) presented short-term site fidelity and 17 individuals (63.0%) presented long-term site fidelity, with a maximum recapture interval of 12 years. Lagged identification rates showed that five individuals (SE = 2) remained in the area for 32 days (SE = 108 days) before leaving and not returning during the same year. Seven individuals were seen both in Madeira and the Canaries (catalogue comprising 51 individuals), three of which were identified multiple times in both archipelagos, with a minimum of 43 days between consecutive sightings.
5. This information combined with the fact that this species is commonly sighted accompanied by calves and feeding in both archipelagos highlights the ecological importance of this area for Bryde's whales. This should be taken into consideration by policymakers when implementing conservation measures, where coordination of effort among countries is needed. This study also reinforces the value of using data from platforms of opportunity and of making photographic data open access.