Intrafollicular and systemic serotonin,oestradiol and progesterone concentrations in cycling mares

The hypothesis that a local serotonergic network might also exist in the follicle of mares remains poorly documented, with exception for humans and laboratory species. For this reason, the aim of the present study was to clarify this possibility, investigating intrafollicular serotonin concentrations of the cycling mare at ovulation time. Sixty ovaries collected from 30 clinically healthy mares of slaughterhouse meat production with clinically normal reproductive tracts after slaughtering were evaluated. Blood samples were taken prior to sacrifice. Follicles were classified in three categories in relation to size, as small (20–30 mm), medium (31–40 mm) and large (>41 mm), and the follicular fluid samples were extracted from each follicle. Intrafollicular and systemic serotonin (5‐HT), oestradiol‐17β (E₂) and progesterone (P₄) were determined by means of enzyme‐linked immunosorbent assay and RIA, respectively. Intrafollicular 5‐HT, E₂ and P₄ concentrations were higher than systemic ones (p < .05). 5‐HT concentrations increased in larger compared to medium follicles, without differences compared to small size follicles (p < .05). 5‐HT and E₂ (r = .79) and 5‐HT and P₄ (r = .79; p < .05) were positively correlated. 5‐HT and P₄ concentrations in follicular fluid increased progressively with the increase in follicular size (p < .05). Follicle diameter and E₂ (r = .85) and P₄ (r = .68) were correlated (p < .05). Since serotonin interacts with steroids, its role on steroidogenesis during growth of the dominant follicle may be suggested.     

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.     

High lysyl oxidase expression is an indicator of poor prognosis in dogs with cutaneous mast cell tumours

Mast cell tumour (MCT) is one of the most frequent skin tumours in dogs. Due to their unpredictable biological behaviour, MCTs often cause several therapeutic frustrations, leading to investigation regarding prognostic markers. Lysyl oxidase (LOX) is an enzyme that promotes extracellular matrix stability and contributes to cell migration, angiogenesis and epithelial-mesenchymal transition. Its expression positively correlates with poor prognoses in several human and canine mammary cancers. The aim of this study was to characterise the immunohistochemical expression of LOX in MCT samples and compare it with histological grading and post-surgical survival. Twenty-six tumours were submitted to immunohistochemistry for LOX expression evaluation. All samples were positive for LOX, with variable percentages of cytoplasmic and nuclear positivity. Cytoplasmic positivity was significantly higher in high-grade MCTs (P = .0297). Our results indicate that high expression of cytoplasmic LOX in neoplastic mast cells is an indicator of poor prognosis for canine cutaneous MCTs.     

Photoprotection of vitamins in skimmed milk by an aqueous soluble lycopene-gum Arabic microcapsule

Riboflavin (Rf)-mediated photosensitized degradation of vitamins A and D3 in skimmed milk under illumination with a white fluorescence lamp was studied by using the HPLC technique. The photosensitized degradation of both vitamins followed first-order kinetics, and the temperature effect on the observed photodegradation rate constant allowed the determination of the activation energy Ea as being 4 and 16 kcal/mol for vitamins A and D3, respectively. The addition of lycopene microencapsulated by spray-drying with a gum arabic-sucrose (8:2) mixture (MIC) produced a reduction of ca. 45% in the photosensitized degradation rate of both vitamins. Front-face fluorescence experiments showed the same photoprotection factor in the degradation of Rf itself, indicating that the photodegradation mechanism involved Rf-mediated reactive species, such as the excited triplet state of Rf, 3Rf*, and/or singlet molecular oxygen, 1O2. The interaction of both 3Rf* and 1O2 with MIC was evaluated in aqueous solutions by using laser-induced time-resolved absorption or emission spectroscopy, and the contribution of an inner-filter effect in the presence of MIC in skimmed milk was evaluated by diffuse reflectance spectroscopy. The main operating mechanism of photoprotection is due to the deactivation of 3Rf* by the proteic component of gum arabic; thus, gum arabic based microcapsules could be used to improve the photostability of milk during its storage and/or processing under light.     

Lectin from Phaseolus acutifolius var. escumite: chemical characterization, sugar specificity, and effect on human T-lymphocytes

Purification of the lectin from Phaseolus acutifolius var. escumite was achieved by affinity chromatography on a column containing glutaraldehyzed membranes from blood group O erythrocytes. The lectin is a tetrameric glycoprotein of 121 kDa with 10% of sugar by weight composed by four subunits of 30 kDa as determined by SDS-PAGE. The lectin is composed of four isolectins as determined by ion-exchange chromatography on a mono-S column. The lectin and its isolectins showed identical NH2 terminal residues (ANDLSFNFQR FNETN) with homology to the PHA leucoagglutinin-precursor. Peptide mass fingerprint from each lectin isoform determined from tryptic peptides by MALDI-TOF (matrix assisted laser desorption ionization-time-of-flight) showed differences among subunits, thus suggesting microheterogeneity in their amino acid sequences or different glycosylation patterns. The lectin and its four isolectins agglutinated erythrocytes without serological specificity and showed mitogenic activity on human leukocytes; moreover, the main effect was rather toward CD8+ than to CD4+ human peripheral lymphocytes. The lectin from escumite was not inhibitable by simple sugars; however, the specificity of the lectin and its isoforms was mainly addressed toward galactose residues present in bi- or triantennary N-acetyllactosamine-type glycans.     

The efficacy of a petroleum spray oil against Aphis gossypii Glover on cotton. Part 1: mortality rates and sources of variation

Petroleum spray oils (PSOs) kill insect pests on contact, and the composition of modern PSOs is substantially different from the ones introduced earlier. The effects of direct application of a new nC24 PSO on the cotton aphid, Aphis gossypii Glover, were therefore determined. This covered not only aphid mortality rates but also the way in which the oils affected aphid behaviour at the time of contact with the oil. Direct application of the nC24 oil proved to be highly effective in controlling A. gossypii at a range of concentrations between 1 and 10% v/v. The oil killed cotton aphids quickly, with most of the mortality occurring within the first 10 min of spraying. The fast killing action of the oils prevented any behavioural responses by the aphids. Aphids killed by the oils became flaccid and their legs and antennae extended horizontally relative to the body axis. With time, their cuticle became very shiny and began to darken. The quick death of the aphids suggested a contact mode of action of the oils, an interpretation supported by the lack of any negative effect on aphids not initially reached by the oils. However, those aphids not hit by the oils, but that subsequently encountered oil-treated areas when they moved elsewhere, also died, indicating that the oil deposits are also toxic to the aphids. The mode of action of the oil thus seems to be versatile, and the means by which it kills the aphids may be more complex than anoxia, which is the widely claimed mechanism attributed to PSOs. This oil now needs to be tested for any possible indirect effects on the cotton aphid (e.g. through its host-plant acceptance behaviour). The implications of the present findings for cotton aphid control and assessment of PSO efficacy in the field are discussed.