In vitro and in vivo stability of caffeic acid phenethyl ester, a bioactive compound of propolis

The in vitro biochemical stability of caffeic acid phenethyl ester in rat and human plasma was investigated and compared with the stability of other caffeic acid esters (chlorogenic acid and rosmarinic acid). The incubation of the compounds in rat plasma for up to 6 h showed that caffeic acid phenethyl ester, but not the other compounds, was hydrolyzed, whereas human plasma did not affect the stability of all the assayed compounds. The products in rat plasma were caffeic acid and an unknown compound, which was identified by mass spectrometry as caffeic acid ethyl ester, produced by transesterification in the presence of ethanol used as vehicle for standard compounds. Specific inhibitors of different plasma esterases allowed the identification of a carboxylesterase as the enzyme involved in the metabolism of caffeic acid phenethyl ester. The oral administration in rats of caffeic acid phenethyl ester in the presence of both ethanol and 2-(2-ethoxyethoxy)ethanol gave rise to a dramatic increase of caffeic acid, as well as low levels of caffeic acid phenethyl ester, caffeic acid ethyl ester, and caffeic acid 2-(2-ethoxyethoxy)ethyl ester, in urine collected within 24 h after treatment. These results suggest that caffeic acid phenethyl ester is hydrolyzed also in vivo to caffeic acid as the major metabolite and that its biological activities should be more properly assayed and compared with those of caffeic acid, its bioactive hydrolysis product. Moreover, alcohols should be carefully used in vivo as solvents for caffeic acid phenethyl ester, since they can give rise to new bioactive caffeic acid esters.     

Phytodecontamination of the Endocrine Disruptor 4-Nonylphenol in Water Also in the Presence of Two Natural Organic Fractions

The objective of this study was to assess the removal of the endocrine disruptor 4-nonylphenol (NP) at a concentration of 1?mg?L^?1 by ryegrass and radish during germination and growth. The decontamination process was evaluated in water only or in water containing two organic fractions, a soil humic acid (HA) and a river natural organic matter (NOM) at concentrations of 10 and 200?mg?L^?1. The addition of these fractions aimed to simulate the organic content of real aqueous systems. At the end of germination and growth, residual NP was measured by chromatographic analysis. Also, NP phytotoxicity was evaluated during germination. In germination experiments, NP in water only was not toxic for ryegrass and radish which removed, respectively, 37 and 51?% of the initial NP added. In water added with HA or NOM at both doses, in general, NP did not influence or inhibited seed germination. Both doses of HA in water promoted the removal of NP by germinating seeds, whereas NOM exerted differentiated results in the two species as a function of the dose applied. After 2?days of growth, in all treatments both species almost completely removed NP and accumulated a very little fraction of product. This study demonstrated that both ryegrass and radish possess a relevant capacity to remove the endocrine disruptor NP from water also in the presence of different organic fractions, thus suggesting their use in the decontamination of real aquatic systems.     

Extenders modify the seminal plasma ability to minimize freeze‐thaw damage on ram sperm

Seminal plasma (SP) proteins interact with sperm plasma membrane (PM) modulating its functionality. It has been shown that SP proteins can reverse the damage caused by freeze‐thaw; however in these studies, SP has been added to washed sperm (i.e., cells depleted from homologous SP and extender). The aim of the current study was to assess whether the egg yolk‐based extender (EY) modifies SP ability to ameliorate sperm parameters in frozen‐thawed ram spermatozoa. Ejaculates were diluted in EY or soybean lecithin‐based extender (SL) and evaluated before and after freezing to measure the cell damage according to the extender. Even when all classical parameters decreased after freezing, as expected (p < .05), there was no effect of the extender. SP treatment was applied after freeze‐thaw. Sperm were incubated with SP (20% v/v) in the presence of either EY or SL, and sperm parameters were assessed after thawing compared with the same treatments after Percoll sperm selection (washed). Treatments with 20% SP improved sperm total and progressive motility compared with controls regardless of washing and extender (p < .05); however, washed sperm showed higher percentage of total sperm motility compared with those unwashed (p < .05). Moreover, treatment with 20% SP showed significantly higher percentages of PM integrity, sperm with intact acrosomes, integrity of chromatin and non‐capacitated sperm in samples diluted with EY when washed before treatment compared with the other conditions (p < .05). It was concluded that the presence of the extenders and particularly egg yolk alters the SP capacity to reduce the cryodamage.     

Properties of dissolved organic matter in forest soils: Influence of different plant covering

The nature of different forest plant litters and the effect of their degradation processes on the properties of soil dissolved organic matter is a very important environmental issue in protected areas such as the “Natural Oriented Reserve Bosco delle Pianelle” in Southern Italy, Chemical and spectroscopic techniques were used to evaluate the influence of four different tree covers on the properties of dissolved organic matter (DOM) isolated from litter and surface soil layers. Fourier transform infrared spectroscopy analysis showed that all litter samples feature a prevalent aromatic character. Ligninolitic components of litters produced by Pinus halepensis L. and a mixture of Quercus trojana Webb. and Q. ilex L. were more degraded than those produced by either Q. ilex or a mixture of Carpinus betulus L. and C. orientalis Miller. The larger C/N ratios of the former litters might account for the greater degradation. Further, a lesser lignin decomposition was shown also for the latter samples, likely due to adverse topographical factors, such as less intense sunlight and greater moisture content. Spectroscopic analysis of litter DOM showed that easily degraded components, such as water-soluble compounds and carbohydrates, were preferentially degraded resulting in DOM that was enriched in lignin-derived compounds. Fluorescence analysis data of all litter DOM samples showed the occurrence of fluorescent units qualitatively similar regardless of plant covering, which suggests that in all cases simple structural components of low-molecular weight and small degree of aromatic polycondensation and content of conjugated chromophores are present. Soil DOM featured several numerous fluorescent units that differ as a function of the parent litter and/or its decomposition processes. Along the soil profiles water-soluble aliphatic compounds of low-molecular weight are found down to the deepest layer, whereas aromatic–ligninolitic compounds are decreasing with depth, probably because of their adsorption by mineral soil components. A greater amount of aromatic units, likely lignin-derived compounds, was found in the DOM from P. halepensis L., which suggested a more extended lignin decomposition. Further, with increasing soil depth, DOM was characterized by a decrease of low-molecular weight organic molecules and lower degree of humification.     

Clinicopathological features and outcome for dogs with mast cell tumors and bone marrow involvement

BACKGROUND: Mast cell tumors (MCTs) with bone marrow (BM) involvement are poorly documented in dogs and are associated with a poor prognosis. Successful treatment strategies have not been described. HYPOTHESIS: Clinicopathologic findings of affected dogs are not specific. Administration of lomustine or imatinib is beneficial. ANIMALS: Fourteen dogs with MCT and BM involvement. METHODS: Clinical and laboratory evaluations were performed in each dog on admission and during follow-up. All dogs received prednisone. Additionally, 8 dogs received lomustine and 3 dogs received imatinib. Imatinib was administered if tumor-associated tyrosine kinase KIT was aberrant. RESULTS: On admission, 11 dogs had a single cutaneous nodule and 3 dogs had multiple nodules. Involvement of regional lymph nodes, liver, or spleen was observed in each dog. BM infiltration with mast cells (MCs) was observed in all dogs. On CBC, nonregenerative anemia, leukopenia, or thrombocytopenia was common. Four dogs had circulating MCs. Increased alkaline phosphatase or alanine transferase activity was observed in 12 and 10 dogs, respectively. Treatment with lomustine induced partial remission in 1 of 8 dogs. Median survival time was 43 days (range, 14-57). Dogs on imatinib experienced complete remission. Two dogs survived for 117 and 159 days, and the third was alive after 75 days. Dogs treated symptomatically did not improve and were euthanized after 1, 14, and 32 days. CONCLUSIONS AND CLINICAL IMPORTANCE: A combination of clinical and laboratory evaluation helps in identifying dogs with MCT and BM infiltration. Administration of lomustine is not helpful in affected dogs. The beneficial effect of imatinib warrants further investigation.     

Soil and compost humic fractions regulate the response of Sclerotinia sclerotiorum to exogenously added allelochemical compounds

Allelochemical compounds released by plants to signal their presence and needs interact in soils with very important macromolecules, such as humic acids (HAs), which are able to modulate the ultimate effects on target organisms. Most of the available studies on plants and microorganisms report the effects of allelochemicals or those of humic fractions, separately. In this study, we investigated the combined activity of these two types of compounds on the soil-resident fungus Sclerotinia sclerotiorum. Thus, ferulic acid (FA), caffeic acid (CA), benzoic acid (BA), salicylic acid (SA), gallic acid (GA) and phthalic acid (PA), exogenously applied to the fungal growth medium, were tested both alone and in combination with a soil HA (SHA) and a compost HA (CHA). The two HAs were also tested alone on the fungus. When the allelochemicals were applied alone, only FA, BA and SA evidenced a significant inhibition of mycelial growth, whereas FA, BA and CA increased the number of sclerotia formed. The two HAs alone reduced the early growth of the fungus and markedly stimulated sclerotia formation. A significant attenuation or, in some cases, suppression of the allelochemical effect on mycelial growth was caused by the coexistence in the medium of the allelochemical and each HA, especially CHA. Moreover, in general, the combinations of HA-allelochemical significantly stimulated sclerotia formation, with respect to the sole allelochemical, but decreased it with respect to HA alone. Thus, investigations on the response of fungi to plant-released allelochemicals should not exclude interactive aspects of these compounds with ubiquitous coexisting humic macromolecules.     

Electroejaculation Increases Low Molecular Weight Proteins in Seminal Plasma Modifying Sperm Quality in Corriedale Rams

This study was conducted to evaluate the effect of seminal collection method (artificial vagina or electroejaculation) on the protein composition of seminal plasma and sperm quality parameters in Corriedale rams. To address this question, we assessed the effect of seminal collection method on motility, plasma membrane integrity and functionality, mitochondrial functionality and the decondensation state of nuclear chromatin in sperm cells. Volume, pH, osmolarity, protein concentration, total protein content and protein profile using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE) and 2‐D polyacrylamide electrophoresis of seminal plasma collected with artificial vagina and electroejaculation were also analysed. The main findings from this study were that ejaculates obtained with electroejaculation had (i) a higher number of spermatozoa with intact plasma membrane and functional mitochondria and (ii) a higher proportion of seminal plasma, total protein content and relative abundance of low molecular weight proteins than ejaculates obtained with artificial vagina. Five of these proteins were identified by mass spectrometry: binder of sperm 5 precursor; RSVP14; RSVP22; epididymal secretory protein E1 and clusterin. One protein spot with molecular weight of approximately 31 kDa and isoelectric point of 4.8 was only found in the seminal plasma from electroejaculation.     

A pest alien invasion in progress: potential pathways of origin of the brown marmorated stink bug Halyomorpha halys populations in Italy

Journal of Pest Science - The brown marmorated stink bug, Halyomorpha halys, is an agricultural and household pest native of far East. In the last years, it has spread to and established in...     

Properties of humic acids in Mediterranean forest soils (Southern Italy): influence of different plant covering

The chemical and spectroscopic properties of humic acids (HAs) isolated from four litters and their corresponding underlying soils at three depths in a protected forest area in Southern Italy were investigated as a function of four different plant coverings: Quercus ilex L., mixed Carpinus betulus L. and C. orientalis Mill., Pinus halepensis L., and mixed Quercus trojana Webb. and Q. ilex L.. The forested site is a part of a calcareous plateau, characterized by homogeneous soils classified as Eutric Cambisols associated with Calcic Luvisols. The changes in the composition of HAs with soil depth have been evaluated on the basis of chemical (elemental and COOH groups) and spectroscopic analyses (E₄/E₆ ratio and FT IR spectra), and lignin-derived CuO oxidation products. A different distribution of the main elements was found in the various HAs which is apparently related to the type of humic acid-precursor biomolecules in the parent litters. The HAs isolated from soils under Q. ilex and mixed Carpinus species showed a slight increase in the C/H ratio and COOH content downward the soil profile, suggesting increasing aromatic polycondensation and humification degree with depth. On the contrary, no trend was observed for HAs from soils under Pinus halepensis L. and mixed Quercus species, indicating a partial incorporation of residues deriving from litter degradation into these HAs. Further, the content of lignin-derived phenols was higher in Pinus halepensis L. and mixed Quercus species layers, with the same trend measured for the corresponding HAs, thus confirming a lignin contribution related to the lignin type of plant covering.