Factors affecting the contents of iridoid oleuropein in olive leaves (Olea europaea L.)

In this study, for the first time, the impact of the genetic factor on the contents of oleuropein in olive leaves was not only evaluated but the influence exerted by the color/age of leaves (green, green-yellowish, and yellow) and the collecting period (spring or autumn) was also evaluated. A repetitive high-resolution gas chromatographic quantitation method and an accurate high-performance liquid chromatographic method were developed. These analytical methods gave results showing a highly linear relationship. Samples of olive leaves were taken from seven major Italian olive cultivars, such as Dritta, Leccino, Caroleo, Coratina, Castiglionese, Nebbio, and Grossa di Cassano. Such a vegetal raw material could actually be exploited for recovering oleuropein, considered to be a high-added value molecule. This could be converted into hydrxytyrosol, a compound known to possess strong bioactive properties. Olive leaves showed considerable contents of oleuropein, which with some cultivars were even higher with respect to those present in the corresponding olive fruits (reported in the literature). The amounts of oleuropein in the collected leaves were markedly modified by the color/age and genetic factors, whereas meaningless variations were ascribable to the quantitation method and the collecting period factors. Various chemometrics, applied to the obtained analytical data, appeared to be effective in discriminating the samples on the basis of the above-examined experimental factors, thus confirming how these should be taken into account in future industrial recovery of oleuropein from olive leaves.     

Chromogranin- and Somatostatin-containing Neuroendocrine Cells in the Porcine Uterus An Immunocytochemical Study

Endocrine cells scattered in organic mucosae were defined "Neuroendocrine" (NE) cells because they constitute a section of the Diffuse Neuroendocrine System (DNES). Such cells have never been evidentiated in the normal endometrium. By means of histochemical and immunohistochemical techniques, NE cells, some of which contain the hormone somatostatin, are described in the glandular epithelium of the uterine horn in non-pregnant, non-castrated, young and adult sows. As is known, the uterine horn is the organ of pregnancy in the pig. The localization, distribution and morphology of the uterine NE and somatostatin-containing cells are reported and the importance of their function, in the pregnant and non-pregnant porcine uterus, is discussed.     

Presence and Distribution of Urocortin and Corticotrophin‐Releasing Hormone Receptors in the Bovine Thyroid Gland

Urocortin (UCN), a 40 amino acid peptide, is a corticotrophin‐releasing hormone (CRH)‐related peptide. The biological actions of CRH family peptides are mediated via two types of G‐protein‐coupled receptors, CRH type 1 (CRHR1) and CRH type 2 (CRHR2). The aim of this study was to investigate the expression of UCN, CRHR1 and CRHR2 by immunoprecipitation, Western blot, immunohistochemistry and RT‐PCR in the bovine thyroid gland. Immunoprecipitation and Western blot analysis showed that tissue extracts reacted with the anti‐UCN, anti‐CRHR1 and anti‐CRHR2 antibodies. RT‐PCR experiments demonstrated that mRNAs of UCN, CRHR1 and CRHR2 were expressed. UCN immunoreactivity (IR) and CRHR2–IR were found in the thyroid follicular and parafollicular cells and CRHR1‐IR in the smooth muscle of the blood vessels. These results suggest that a regulatory system exists in the bovine thyroid gland based on UCN, CRHR1 and CRHR2 and that UCN plays a role in the regulation of thyroid physiological functions through an autocrine/paracrine mechanism.     

Role of soil properties in sewage sludge toxicity to soil collembolans

Soil properties are one of the most important factors explaining the different toxicity results found in different soils. Although there is knowledge about the role of soil properties on the toxicity of individual chemicals, not much is known about its relevance for sewage sludge amendments. In particular little is known about the effect of soil properties on the toxicity modulation of these complex wastes. In addition, in most studies on sewage sludges the identity of the main substances linked to the toxicity and the influence of soil properties on their bioavailability remains unknown.In this study, the toxicity of a sewage sludge to the soil collembolan Folsomia candida was assessed in nine natural soils from agricultural, grassland and woodland sites, together with the OECD soil. Correlations between the relative toxicity of sludge for collembolans in the different soils and their physical and chemical soil properties were assessed in order to identify the main compounds responsible for the effects observed. Furthermore, the relationships between the toxic effects to collembolans and water-soluble ions released by sludge, pH and electric conductivity were also assessed, together with the modulating effects of soil properties.Sludge toxicity was directly linked to the water extractable ammonium, which explained most of the mortality of the collembolans, and part of the inhibition of reproduction. For the last endpoint, nitrite also contributed significantly to the inhibition observed. The varied levels in water extractable ammonium in the different soils at equal dosages seem to be, in turn, modulated by some soil properties. Higher organic carbon contents were associated with lower toxicity of sludge, both for survival and reproduction, probably related to its higher ammonium sorption capacity. In addition, for reproduction, increasing the C/N ratio and pH appeared to increase the toxicity, probably due to both the greater difficultly in nitrification and the known unsuitability of alkaline soils for this species.     

Structural elucidation of phototransformation products of azimsulfuron in water

The photodegradation of the sulfonylurea herbicide azimsulfuron, N-[[(4,6-dimethoxypyrimidin-2-yl)amino]carbonyl]-1-methyl-4-(2-methyl-2H-tetrazole-5-yl)-1H-pyrazole-5-sulfonamide (AZS), was studied in water at different wavelengths and in the presence of photocatalysts. AZS was rapidly degraded by UV light, affording three photoproducts. The main product, accounting for about 70% of photodegraded herbicide, was identified as 6-amino-5-[(4,6-dimethoxypyrimidin-2-yl)methylamino]-1,5,6,8-tetrahydro-7-oxa-8lambda(6)-tia-1,2,5,6-tetraza-azulen-4-one (ADTA) by single-crystal X-ray diffraction. With simulated sunlight irradiation, the reaction was slower and 2-amino-4,6-dimethoxypyrimidine (DPA) and 1-methyl-4-(2-methyl-2H-tetrazole-5-yl)-1H-pyrazole-5-sulfonamide (MPS), arising from a photohydrolytic cleavage of the sulfonylurea bridge, were the only byproducts observed. The reactions followed first-order kinetics. The addition of dissolved organic matter (DOM) did not modify significantly the AZS photodegradation rate. The presence of Fe2O3 accelerated more than twice the reaction rate affording two major products, DPA and MPS, together with minor amounts of N-[[(5-hydroxy-4,6-dimethoxypyrimidin-2-yl)amino]carbonyl]-1-methyl-4-(2-methyl-2H-tetrazole-5-yl)-1H-pyrazole-5-sulfonamide (AZS-OH). The greatest degradation rate was detected in the presence of TiO2. Only the photohydroxylation product AZS-OH was observed, which was transformed rapidly into oxalic acid.     

On the origin of artichoke and cardoon from the <Emphasis Type="Italic">Cynara</Emphasis> gene pool as revealed by rDNA sequence variation

The evolutionary history of artichoke and cultivated cardoon and their relationships to wild allies of the genus Cynara are not fully understood yet. To try resolve the evolutionary patterns leading to the domestication of these two crops, a study of molecular evolution was undertaken. The species C. cardunculus, including artichoke, cultivated and wild cardoon, together with four wild Cynara species were taken into consideration. Internal (ITS) and external (ETS) rDNA transcribed spacers were used as markers of nuclear genome, the psbA-trnH spacer as a marker of chloroplast genome. Sequences were analysed using phylogenetic analysis packages. Molecular data indicate that the whole genus is quite recent and that the domestication of artichoke and cultivated cardoon, crops diverging for reproduction system and use, are independent events which diverge in time and space. As for wild Cynara species, an evolutionary pattern consistent with their present geographical distribution was hypothesized in relation to the climatic changes occurring in the Mediterranean during the last 20 millennia: C. humilis and C. cornigera appeared to have differentiated first, C. syriaca and C. baetica were differentiated in a second period, while C. cardunculus showed to be the most recent and plastic species. The high plasticity of C. cardunculus has not only allowed its nowadays wide distribution, but has also given the potential for domestication. Dedicated to the memory of Richard Neville Lester (1937–2006) who greatly contributed to the understanding of the evolution, domestication, and genetic resources of eggplants as well as to the biosystematics and taxonomy of allied species. IGV Publication N. 78 of the Institute of Plant Genetics, CNR     

Response of rainbow trout gill (Na++K+)-ATPase and chloride cells to T3 and NaCl administration

With the aim of comparing the effects of oral T₃ and NaCl administration on trout hypoosmoregulatory mechanisms, three groups of rainbow trout (Oncorhynchus mykiss Walbaum) held in freshwater (FW) were fed a basal diet (C), the same diet containing 8.83 ppm of 3,5,3-triiodo-L-thyronine (T₃) (T) or 10% (w/w) NaCl (N) respectively for 30 d. They were then transferred to brackish water (BW) for 22 d and fed on diet C. Gill (Na⁺+K⁺)-ATPase activity and its dependence on ATP, Na⁺ and pH, number of gill chloride cells (CC), serum T₃ level as well as fish growth, condition factor (K) and mortality were evaluated. During the FW phase, as compared to C trout, T trout showed a two fold higher serum T₃ level, had unchanged gill (Na⁺+K⁺)-ATPase activity and increased CC number, whereas N trout showed higher gill (Na⁺+K⁺)-ATPase activity and CC number. At the end of the experiment the enzyme activity was in the order T>N>C groups and all groups showed similar CC number. Both treatments changed the enzyme activation kinetics by ATP and Na⁺. A transient increase in K value occurred in N group during the period of salt administration. In BW, T and N groups had higher and lower survival than C group respectively. Other parameters were unaffected by the treatments. This trial suggests that T₃ administration promotes the development of hypoosmoregulatory mechanisms of trout but it leaves the (Na⁺+K⁺)-ATPase activity unaltered till the transfer to a hyperosmotic environment.     

Effect of thinning on above-ground biomass accumulation in a Douglas-fir plantation in southern Italy

We investigated the effects of a long-term thinning experiment on the distribution of above-ground biomass of Douglas-fir(Pseudotsuga menziesii [Mirb.]Franco var. menziesii) in a plantation in southern Italy.Allometric equations were used to estimate biomass and partitioning to stem and crown compartments. Variation in biomass stock estimated with allometric equations were evaluated according to seven thinning treatments: geometric-systematic(1 row every 3), selective(light-moderate-heavy), mixed systematic-selective(1 row every 4, 1 row every 5), unthinned(control). Over the experimental period of 13 years, current annual increments of carbon were lower(3.4 Mg ha~(-1)year~(-1)) in control plots than in treated plots. At age 30, plots subjected to light selective thinning showed higher values of above-ground biomass(249.7 Mg ha~(-1)). The biomass harvested with this treatment was 29.3 Mg ha~(-1), and the mean annual increment of carbon over 13 years was 4.8 Mg ha~(-1). Our results showed that light thinning stimulated increase in carbon stock, with a minimal loss of carbon during the treatment and a current annual increment of carbon higher than in control sub-plots and sub-plots thinned using systematic methods. This treatment yielded least carbon emissions and we affirm it has discrete global warming mitigation potential.     

Fenhexamid adsorption behavior on soil amended with wine lees

The adsorption of fenhexamid (FEN) [N-(2,3-dichloro-4-hydroxyphenyl)-1-methylcyclohexanecarboxamide] on vineyard soil amended with wine lees (WL) produced by vinery was studied. The adsorption extent depends on WL fraction. The addition of the centrifuged solid lees (SWL) increases the FEN adsorption on soil. Most likely, the organic insoluble fraction formed mainly by dead fermentation yeasts is responsible for the observed increase. The adsorption measured on some deactivated yeasts of wine fermentation shows that Saccharomyces cerevisiae are the most active in FEN retention. On the other hand, the soil amendment with whole WL decreases considerably the fungicide adsorption. This opposite effect may be the result of FEN hydrophobic bonds with the dissolved organic matter of lees that keeps fungicide in solution. This hypothesis is substantiated by the increased FEN solubility in the supernatant of centrifuged wine lees (LWL). The results of soil column mobility confirm that the elution with LWL increases the mobility of FEN in soil.     

Presence and Distribution of Urocortin and its Receptors in the Epididymis of Alpaca (Vicugna pacos)

Urocortin 1 (UCN) is a 40‐amino acid peptide belonging to the corticotrophin‐releasing hormone (CRH) family. The biological effects of this peptide are modulated by binding two G‐coupled receptors named CRH receptor 1 (CRHR1) and CRH receptor 2 (CRHR2). CRHR2 has high affinity for UCN. The aim of the present study was to investigate the presence and distribution of UCN, CRHR1 and CRHR2 in the epididymis of the South America camelid Alpaca (Vicugna pacos) by Western blotting analysis and immunohistochemistry. Tissue extracts of the organ reacted with the anti‐UCN, anti‐CRHR1 and anti‐CRHR2 antibodies, recognizing in all the cases a single specific protein band. UCN‐ and CRHR2‐immunoreactivities (IRs) were found in the cytoplasm of the principal cells (PCs) of the caput epididymis. A prevalent supranuclear localization of granular‐shaped positive material was observed. CRHR1‐IR was observed in the fibromuscular stromal cells encircling the tubules and in the smooth musculature of the blood vessels throughout the three epididymal segments. In addition, in the cauda, CRHR1‐IR was observed in some apical epithelial cells (ACs) which were morphologically similar to apical mitochondria‐rich cells (AMRCs). These results suggest that UCN, CRHR1 and CRHR2 are expressed in the alpaca epididymis and that CRH‐related peptides might play multiple roles in maturation and storage of spermatozoa.