The effect of extraction process on polyphenol content, triterpene composition and bioactivity of yellow birch (Betula alleghaniensis Britton) extracts

Crude ethanol extracts from different tissues of Betula alleghaniensis: wood, inner and outer bark, foliage, and twigs were evaluated for their nitric oxide (NO)-radical scavenging activities and inhibitory effects on the production of NO in LPS/INFγ-stimulated RAW 264.7 macrophages. As a renewed interest in plant-derived drugs has led to an increased need for efficient extraction methods, ultrasonic-assisted extraction (UAE) was investigated and compared with conventional maceration. The lower energy consumption by the UAE process compared to maceration determined in our study qualifies also this extraction as an environmentally friendly process. Our results indicate that the foliage extract has the most potent radical scavenging capacity while wood and twigs extracts exert the highest inhibitory effects on the production of NO in LPS/INFγ-stimulated RAW 264.7 macrophages. Extracts obtained by UAE present lower cytotoxic activity on RAW cells. Our results also demonstrate that ultrasounds help to selectively extract the bioactive molecules from foliage, twigs and wood, which inhibit the production of NO by macrophages. The highest total phenol content has been determined for the inner bark extracts and the flavonoids are the major phenolics present in foliage extracts. Poor correlations determined between the total phenols and radical scavenging capacity of the extracts indicates to the synergistic or antagonist effects of molecules present, various polyphenols and triterpenes being identified in the extracts studied in this research.     

Genotype by environment interactions in barley (<Emphasis Type="Italic">Hordeum</Emphasis><Emphasis Type="Italic"> vulgare</Emphasis> L.): different responses of landraces,recombinant inbred lines and varieties to Mediterranean environment

Genotype-by-environment interactions (GEIs) can affect breeding progress because they often complicate the evaluation and selection of superior genotypes. This drawback can be reduced by gaining insights into GEI processes and genotype adaptation. Here, we have studied the GEIs over a set of 24 barley genotypes that were grown across six environments (location-by-year combinations) in Sardinia, Italy. Three groups of genotypes were analysed: barley landraces (LANs), recombinant inbred lines (RILs), and commercial varieties (VARs). The additive main effects and multiplicative interaction (AMMI) model was used for data analysis, and results evidenced no significant differences in grain yield averages for the 24 genotypes. However, there was a relevant GEI for yield mainly between two of the six environments (one characterised by warm pre-anthesis period and high spring rainfalls, and the other characterised by opposite features) and two groups of genotypes (VAR and LAN). Moreover, a negative trade-off between yield levels of genotypes was seen when the barley genotypes were grown in the contrasting environments. Overall, intermediate GEI levels were seen for RILs in comparison to LANs and VARs, and some of the RILs provided valuable yield levels (e.g. RILs 23 and 52). The results thus show the potential usefulness of LANs as a genetic resource for breeding, e.g. to obtain genotypes adapted to Mediterranean environments, such as the RILs analysed in this study. Most of the actual work was carried out when the first author was a PhD student in ‘Agro-meteorology and ecophysiology of agricultural and forest systems’ and she was affiliated to Dipartimento di Scienze Agronomiche e Genetica Vegetale Agraria, Università degli Studi di Sassari, Via E. de Nicola, Sassari 07100, Italy.     

Identification of non-alkaloid acetylcholinesterase inhibitors from Ferulago campestris (Besser) Grecescu (Apiaceae)

Inhibition of Acetylcholinesterase (AChE) is still considered as a strategy for the treatment of neurological disorders such as Alzheimer's disease. Many plant derived alkaloids (such as galantamine and rivastigmine) are known for their AChE inhibitory activity. Recently, other classes of natural compounds such as terpenoids, sesquiterpene glycosides and coumarins have been studied as new AChE inhibitors, with the aim to discover less toxic compounds compared to alkaloidal ones. The Ferulago campestris roots dichloromethane extract was used for a bioassay-guided fractionation for the search of AChE inhibitors. Three coumarin derivatives (umbelliprenin 1, coladonin 2 and coladin 3), three daucane ester derivatives (siol anisate 4, ferutinin 5 and 1-acetyl-5-angeloyl lapiferol 6), two phenol derivatives (2-epilaserine 7 and epielmanticine 8) and one polyacetylene (9-epoxyfalcarindiol 9) were isolated by the bioassay-guided approach. Their structures were characterized on the basis of spectral methods (1D and 2D NMR, and MS spectroscopy). All the isolated compounds were able to inhibit the AChE (IC₅₀ 1.2–0.1 mM) although at higher doses if compared to galantamine (6.7 μM) measured in the same conditions. The most active compounds were the daucane derivative siol anisate 4 and the epielmanticine 8, with IC₅₀ of 0.172 and 0.175 mM respectively.     

Response of alfalfa (Medicago sativa L.) to single and mixed inoculation with phosphate-solubilizing bacteria and Sinorhizobium meliloti

The objectives of this work were to phenotypically and genetically characterize alfalfa rhizosphere bacteria and to evaluate the effect of single or mixed inoculation upon nodulation and biological nitrogen fixation. Thirty-two strains showed tricalcium phosphate solubilization ability, and two of them caused bigger or equal solubilization halos than the control strain P. putida SP22. The comparison of the 16S ribosomal DNA sequences indicated that these strains are phylogenetically related to Bacillus spp. and Pseudomonas spp. A beneficial effect of both isolates on alfalfa growth was observed in coinoculation assays. Pseudomonas sp. FM7d caused a significant increase in root and shoot dry weight, length, and surface area of roots, number, and symbiotic properties of alfalfa plants. The plants coinoculated with Sinorhizobium meliloti B399 and the Bacillus sp. M7c showed significant increases in the measured parameters. Our results indicating that strains Pseudomonas sp. FM7d and Bacillus sp. M7c can be considered for the formulation of new inoculants.     

Influence of Steroidal Anti-Inflammatory Drugs on Viability and Fertility of Equine Semen

Considering the efficiency of steroidal anti-inflammatory drugs (SAIDs) in the immunomodulation of breeding-induced endometritis and the possibility of using these drugs by intrauterine route instead of the parenteral application, the present study aimed at evaluating the effect of SAIDs added to the semen extender on equine sperm viability and fertility. In experiment 1, 15 SAIDs were individually added to a skim milk-based extender and, based on the results of sperm motility, dexamethasone was the drug of choice for the subsequent trials. The effect of dexamethasone on the viability of fresh and 24-hour cooled semen was investigated in experiment 2. In experiment 3, fertility rate was measured in both post-breeding endometritis-resistant and susceptible mares. Although dexamethasone supplementation caused a premature decrease in sperm total and progressive motility and in sperm velocities (P < .05), no difference was observed for sperm membrane integrities and fertility (P > .05). Based on these results, we can conclude that dexamethasone can be added to equine semen at the time of insemination or before cooling, although its use was not able to increase fertility.     

Resistance to ETEC F4/F18–mediated piglet diarrhoea: opening the gene black box

Diarrhoea, a significant problem in pig rearing industry affecting pre- and post-weaning piglets is caused by enterotoxigenic Escherichia coli (ETEC). The ETEC are classified as per the fimbriae types which are responsible for bacterial attachment with enterocytes and release of toxins causing diarrhoea. However, genetic difference exists for susceptibility to ETEC infection in piglets. The different phenotypes found in pigs determine their (pigs’) susceptibility or resistance towards fimbrial subtypes/variants (F4ab, F4ac, F4ad and F18). Specific receptors are present on intestinal epithelium for attachment of these fimbriae, which do not express to same level in all animals. This differential expression is genetically determined and thus their genetic causes (may be putative candidate gene or mutations) render some animals resistant or susceptible to one or more fimbrial subtypes. Genetic linkage studies have revealed the mapping location of the receptor loci for the two most frequent variants F4ab and F4ac to SSC13q41 (i.e. q arm of 13th chromosome of Sus scrofa). Some SNPs have been identified in mucin gene family, transferring receptor gene, fucosyltransferase 1 gene and swine leucocyte antigen locus that are proposed to be linked mutations for resistance/susceptibility towards ETEC diarrhoea. However, owing to the variety of fimbrial types and subtypes, it would be difficult to identify a single causative mutation and the candidate loci may involve more number of genes/regions. In this review, we focus on the genetic mutations in genes involved in imparting resistance/susceptibility to F4 or F18 ETEC diarrhoea and possibilities to use them as marker for selection against susceptible animals.

     

Nitrate and ammonium nutrition in chicory and rocket salad plants

To evaluate chicory (Cichorium intybus L.) and rocket salad [Eruca vesicaria (L.) Cav.subsp. sativa (Mill.)] capability to use ammonium‐nitrogen (NH₄‐N) even in the absence of nitrate‐nitrogen (NO₃‐N) in the nutrient solution, and the chances they offer to reduce leaf NO₃ content, cultivated rocket and two cultivars of chicory ('Frastagliata’, whose edible parts are leaves and stems, and ‘Clio’, a leaf hybrid) were hydroponically grown in a growth chamber. Three nutrient solutions with the same nitrogen (N) level (4 mM) but a different NH₄‐N:NO₃‐N (NH₄:NO₃) ratio (100:0, 50:50, and 0:100) were used. Rocket growth was inhibited by NH₄ nutrition, while it reached the highest values with the NH₄:NO₃ ratio 50:50. Water and N‐use efficiencies increased in rocket with the increase of NO₃‐N percentage in the nutrient solution. In the best conditions of N nutrition, however, rocket accumulated NO₃ in leaves in a very high concentration (about 6,300 mg kg^‐1 fresh mass). For all the morphological and yield features analyzed, chicory resulted to be quite unresponsive to N chemical forms, despite it took more NO₃‐N than NH₄‐N when N was administered in mixed form. By increasing NO₃‐N percentage in the nutrient solution, NO₃ leaf content increased (5,466 mg kg^‐1 fresh mass with the ratio NH₄:NO₃ 0:100). On average, both chicory cultivars accumulated 213 mg NO₃ kg^‐1 fresh mass with the ratio NH₄:NO₃ 100:0 and, differently from rocket, they showed that by using NH₄ produce can be obtained very low in NO₃ content.     

The Effects of Refrigeration Temperature and Storage Time on Apoptotic Markers in Equine Semen

Evaluation of the damage caused by the sperm preservation process is crucial to improving fertilization rates. The objective of this study was to evaluate the effects of refrigeration temperature (5°C and 15°C) and storage time (0, 12, 24, 48, and 72 hours) on apoptotic markers in equine semen. Membrane phosphatidylserine translocation index, caspase activation index, and DNA fragmentation index were analyzed using epifluorescence microscopy. Analysis of variance was used for statistical analysis, and Tukey test was used to compare means. The significance level was set at P < .05. The results demonstrated that for transport duration shorter than 24 hours, semen quality was maintained when stored at either 5°C or 15°C. A storage temperature of 5°C should be used when it is necessary to transport semen for longer than 24 hours. There was a significant decrease in semen quality after 48 hours of refrigeration.     

Effect of Storage Time and Temperature of Equine Epididymis on the Viability,Motion Parameters,and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time.