Antiproliferative terpenoids from almond hulls (Prunus dulcis): identification and structure-activity relationships

Bioassay-guided fractionation of the EtOAc crude extract from Sicilian almond hulls, a waste material from Prunus dulcis crop, allowed identification of 10 constituents, isolated as pure compounds (1-5, 7, and 10) or unseparable mixtures (5 + 6 and 8 + 9). All compounds were subjected to spectroscopic analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide bioassay on MCF-7 human breast cancer cells. In addition to the main components oleanolic (1), ursolic (2), and betulinic (3) acids, the 2-hydroxy analogues alphitolic (4), corosolic (5), and maslinic (6) acids, as well as the related aldehydes, namely, betulinic (7), oleanolic (8), and ursolic (9), were identified. From a more polar fraction, the beta-sitosterol 3-O-glucoside (10) was also identified. A sample of commercially available betulin (11) was also included in bioassays as further support to a structure-activity relationship study. Betulinic acid showed antiproliferative activity toward MCF-7 cells (GI50 = 0.27 microM), higher than the anticancer drug 5-fluorouracil.     

Effects of after-ripening,stratification and GA3 on dormancy release and on germination of wild asparagus (Asparagus acutifolius L.) seeds

Seeds of wild asparagus (Asparagus acutifolius L.) were treated and compared in this research to investigate seed dormancy class and level involved in this species. Four seed lots were compared: (i) freshly harvested seeds in 2007 (07Fr); (ii) freshly harvested seeds in 2008 (08Fr); (iii) after-ripened (AR) 2007 seeds dry stored in glass jars (ARg); (iv) AR 2007 seeds dry stored in paper bags (ARp). The 07Fr seeds were exposed to (1) chemical scarification combined with gibberellic acid (GA₃) levels (0, 200, 400, and 600 mg L⁻¹) and to (2) 28-day moist stratification at 5 and 23 °C, and two sequences of 5/23 °C combined with 0 and 400 GA₃ mg L⁻¹ levels, and (3) together to the 08Fr and AR seeds were exposed to 56-day moist stratification at 5, 23, or 5/23 °C. With the 08Fr and AR seed lots this last stratification treatment was combined with 0 or 800 GA₃ mg L⁻¹ levels. The dormancy depth of 08Fr (32% germination) was less than 07Fr seeds (2%). The latter after-ripened during dry storage and when stored in glass germinated more (47.5%) than in paper (12%). Stratification for 4 weeks was ineffective in improving germination of 07Fr seeds; when chemically scarified they did not germinate at all. The highest (nearly 70%) and the most rapid and uniform germination were observed for all the lots when they were warm stratified for 56 days. Warm stratification improved germination more than alternate temperature stratification, while cold stratification inhibited germination especially for the 08Fr and ARg lots, thus seeds seem not to have a morphological component to their dormancy. GA₃ only improved germination of 07Fr seeds, at a low rate. A. acutifolius seeds fit the characteristics of a non-deep physiological dormancy.     

SSR markers reveal the uniqueness of olive cultivars from the Italian region of Liguria

Twenty-three important Ligurian olive accessions corresponding to 16 cultivars were studied using 12 SSR markers and 40 Mediterranean cultivars were included in the study in order to investigate the relationships between Ligurian and Mediterranean germplasm. All SSRs produced polymorphic amplifications. One hundred and forty-nine alleles were found in the 63 accessions analysed. Twenty-two alleles were specific to germplasm from Liguria and of these 12 were unique to single cultivars. Heterozygosity and discriminating power calculated in this regional germplasm were high on average (0.70 and 0.74) and not so much lower than the values in the total sample that includes cultivars from different Mediterranean countries (0.77 and 0.88 respectively). No cases of genetic identities were found between Ligurian and Mediterranean accessions. Several cases of homonyms and synonyms within the Ligurian germplasm were explained. Cluster analysis generally revealed a clear discrimination of the profiles from Liguria and Italy with respect to the cultivars from other Mediterranean countries. Only one Ligurian cultivar, “Negrea”, appeared to have a different origin, grouping with the Mediterranean cultivars.     

Estimating clinical chemistry reference values based on an existing data set of unselected animals

In an attempt to standardise the determination of biological reference values, the International Federation of Clinical Chemistry (IFCC) has published a series of recommendations on developing reference intervals. The IFCC recommends the use of an a priori sampling of at least 120 healthy individuals. However, such a high number of samples and laboratory analysis is expensive, time-consuming and not always feasible, especially in veterinary medicine. In this paper, an alternative (a posteriori) method is described and is used to determine reference intervals for biochemical parameters of farm animals using an existing laboratory data set. The method used was based on the detection and removal of outliers to obtain a large sample of animals likely to be healthy from the existing data set. This allowed the estimation of reliable reference intervals for biochemical parameters in Sarda dairy sheep. This method may also be useful for the determination of reference intervals for different species, ages and gender.     

Intrinsic innervation of the horse ileum

This paper describes the morphology and distribution of the enteric nervous system (ENS) cells and fibres immunoreactive for choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene-related peptide (CGRP), NF200 kDa (NF200), and S100 protein. The percentages of subclasses of enteric neurons in the total neuronal population were investigated by the use of anti-PGP 9.5 or anti-NSE antibodies.ChAT-IR myenteric plexus (MP) and submucosal plexus (SMP) neurons were 66 ± 7% and 74 ± 15%, respectively, whereas those cells expressing nNOS-IR were 38 ± 7% and 5 ± 1%, respectively. MP and SMP neurons expressing both phenotypes were also present. SP-IR was expressed by 14 ± 13% of MP and 66 ± 8% of SMP neurons whereas CGRP-IR was observed only in the SMP (43 ± 6%). NF200-IR was expressed by 61 ± 15% and 91 ± 10% of the MP and SMP neurons, respectively. The majority of the CGRP-IR SMP neurons expressed also SP-IR. Almost all SP-IR neurons in both the plexuses were cholinergic. The present study quantifies the main neuronal subpopulations of the ENS of the horse ileum; these data might be utilized to understand the neuronal modifications which occur in several gastrointestinal tract disorders.     

Quantitative determination of dietary lectin activities by enzyme-linked immunosorbent assay using specific glycoproteins immobilized on microtiter plates

An immunoenzymatic method for the quantitative determination of dietary lectin activities employing immobilized glycoproteins was studied. Lectins from wheat germ (WGA), peanut (PNA), and jack bean (ConA) were added to microtiter plates coated with ovalbumin or asialofetuin and quantified by enzyme-linked immunosorbent assay (ELISA) with lectin-specific antibodies. ELISA responses for lectin activity were dose-dependent in the concentration range 30-1000 ng/mL for WGA and 80-1000 ng/mL for both PNA and ConA. Inhibition assays carried out with different saccharides confirmed that the binding of lectins to immobilized glycoproteins was specific. The proposed method is specific and sensitive, allowing the quantitative determination of lectin activities on raw samples by simple dilution of the extracts. Examples of application to wheat germ and roasted peanut extracts are reported.     

A contribution to the environmental biology of mercury accumulation in plants

Samples of six common plant species collected in the old mining areas near Prince George, British Columbia (Canada) and Mount Amiata, Tuscany (Italy) show remarkable similarities in the variation of plant/soil Hg concentration ratio with soil Hg content irrespective of species or other biological differences. In contrast, plants sampled in the geothermally active areas of New Zealand, Hawaii and around Mount St. Helens exhibit more individuality in the concentration ratio to soil Hg relationship, but the relationships are distinctly different from the mine site specimens. This distinction is particularly evident when the same species of Equisetum and Plantago taken from these two different areas are compared. These and other data support the hypothesis that specific local environmental factors strongly influence the accumulation of Hg in plants even when the immediate soil concentrations are the same. Our findings show that some plants contain concentrations of total Hg as high as 5500 to 14000 μg kg^?1 (dw).     

Transgenic plants expressing immunosuppressive dsRNA improve entomopathogen efficacy against Spodoptera littoralis larvae

Journal of Pest Science - Transgenic plants that express double-stranded RNA (dsRNA) targeting vital insect genes have recently emerged as a valuable new tool for pest control. In this study,...     

Physical, Hematological, and Biochemical Responses to Acute Intense Exercise in Polo Horses

The aim of this study was to evaluate the response of physical, hematological, and biochemical parameters after acute intense exercise in polo horses playing in an outdoor international competition. The game consisted of four periods (chukkas) and each period consisted a playing time of 7 minutes. Two matches were played everyday for a week. A total of 12 horses were examined. Each “high-goal” polo horse played one chukka a day for 4 days. Horses were clinically examined the day before the games started and then daily during the 4 days of their participation in the games. During these days, physical examination was performed and blood sample was collected at rest (T0), immediately (T1) after exercise, and after 30 minutes of exercise (T2). Blood samples were analyzed for total cell counts and for determination of creatine kinase, lactate dehydrogenase (LDH), aspartate aminotransferase, lactate, total proteins, calcium, magnesium, phosphorus , and cortisol. Data were evaluated using two-way analysis of variance. Exercise caused significant dehydration (P < .01), mucous membranes congestion, increased heart rate (P < .001), and capillary refill time (P < .001). It also caused increased value of the following parameters: hematocrit (P < .001), red blood cells (P < .001), hemoglobin (P < .001), white blood cells (P < .05), lymphocyte (P < .001), total proteins (P < .001), creatine kinase (P < .05), LDH (P < .01), lactate (P < .001), and cortisol (P < .01), and a decrease in the platelet count (P < .001), calcium (P < .01), phosphorus (P < .001), and magnesium (P < .001). All parameters returned within or near the reference range by 30 minutes postexercise. On the basis of these observations, data were considered indicative of a good response to an acute intense exercise. Moreover statistical results obtained were typical of a mixed aerobic/anaerobic metabolic pathway that is prevailing in this sport.     

Expression of verocytotoxic Escherichia coli antigens in tobacco seeds and evaluation of gut immunity after oral administration in mouse model

Verocytotoxic Escherichia (E.) coli strains are responsible for swine oedema disease, which is an enterotoxaemia that causes economic losses in the pig industry. The production of a vaccine for oral administration in transgenic seeds could be an efficient system to stimulate local immunity. This study was conducted to transform tobacco plants for the seed-specific expression of antigenic proteins from a porcine verocytotoxic E. coli strain. Parameters related to an immunological response and possible adverse effects on the oral administration of obtained tobacco seeds were evaluated in a mouse model. Tobacco was transformed via Agrobacteium tumefaciens with chimeric constructs containing structural parts of the major subunit FedA of the F18 adhesive fimbriae and VT2e B-subunit genes under control of a seed specific GLOB promoter. We showed that the foreign Vt2e-B and F18 genes were stably accumulated in storage tissue by the immunostaining method. In addition, Balb-C mice receiving transgenic tobacco seeds via the oral route showed a significant increase in IgA-positive plasma cell presence in tunica propria when compared to the control group with no observed adverse effects. Our findings encourage future studies focusing on swine for evaluation of the protective effects of transformed tobacco seeds against E. coli infection.