Daily variations of plasma serotonin levels in 2-year-old horses

Serotonin (5-HT) is a neurotransmitter implicated in the regulation of circadian rhythms. Synthesis and release of 5-HT are subjected to daily fluctuations, but there are no data on plasma serotonin levels in 2-year-old horses. Aim of this study was to investigate the daily variations in 5-HT levels in platelet-poor plasma from 2-year-old horses, so as to understand whether plasma 5-HT could be considered as an additional variable to assess the natural adaptability of horses to training. The research was carried out on 6 clinically healthy crossbred mares. Data showed higher levels of platelet-poor plasma 5-HT at 5 p.m. (P < 0.001) than at 1 p.m. and lower values at 5 a.m. (P < 0.001) than at 1 a.m. One-way repeated-measures analysis of variance showed a significant effect of sampling time (P < 0.0001) on plasma 5-HT changes. Results suggest that the increase of 5-HT levels could be related to an enhanced 5-HT output and release, and that their daily variations could be considered as an additional variable to assess the natural adaptability of horses for being trained. Tryptophan levels, the essential amino acid precursor of 5-HT, seemed to hint at slight fluctuations at regular intervals, with the highest amounts at 9 a.m. and at 9 p.m. and lowest at 1 a.m. and at 1 p.m., in agreement with data reported in adult horses. Cortisol levels showed wide fluctuations, with the highest amounts at 1 a.m. and the lowest amounts at 9.00 p.m.     

Capillary electrophoresis for the detection of bilirubin in rat serum

Background: The rat is used often to assess the toxicity of new chemical entities in preclinical drug development. Bilirubin concentration in rat serum is routinely determined by colorimetric methods, but false positive results due to hemolyzed serum or direct interferences by test compounds may occur. Capillary electrophoresis (CE) is an automated method that requires small sample volume and facilitates the direct detection of bilirubin. Objective: The purpose of this study was to evaluate a CE method for detecting bilirubin and albumin‐bound bilirubin in rat serum and to measure potential interference by hemolysis and specific test compounds. Methods: Serum samples from male Sprague Dawley rats (n=20) were used in the study. Results obtained on a Beckman P/ACE MDQ CE instrument equipped with a UV‐detector were compared with those obtained using a colorimetric method on a Hitachi 912 analyzer. Bilirubin standards were used to evaluate the detection and stability of bilirubin in rat serum, and vials with ultrafiltration membranes were used to separate albumin‐bound bilirubin. Intraday and interday coefficients of variation (CV), linearity, and the effects of added hemoglobin and a test compound on CE results were determined. Results: The CE method was capable of detecting bilirubin and albumin‐bound bilirubin in rat serum samples with reproducible results and good accuracy. CVs were <3% and linearity of the CE assay was high (R²=0.9951). Abnormally high bilirubin peaks due to the presence of hemoglobin or the test compound were easily distinguished by means of CE. Conclusion: CE is a good alternative to the colorimetric methods currently used for the determination of bilirubin in rat serum.     

Role of equine herpesviruses as co-infecting agents in cases of abortion, placental disease and neonatal foal mortality

Herpesviral infections frequently occur in horses. The objective of this study was to investigate the possible association of equine herpesviruses (EHV-1, EHV-2, EHV-3, EHV-4, EHV-5) with other causes of abortion, neonatal mortality or placental disorder. Sixty-seven abortions, 22 stillbirths, 14 cases of neonatal foal mortality and 3 cases of placental disease were investigated for infectious and non-infectious causes. Type-specific nested PCR assays and virus isolation were performed to detect EHV infections. A cause of fetal loss or placental disease was reached in 68 out 116 (58.7 %) cases. Twenty-seven cases were positive for EHV, and 22/27 (81.5 %) were positive for EHV-1 (16 neuropathogenic and 6 non-neuropathogenic strains), 4 (14.8 %) for EHV-2 and 3 (11.1 %) for EHV-5. The association between EHV infections and other etiological agents was statistically significant (two sided P?=?0.002). The odds ratio of EHV DNA associated with other diagnoses, especially with bacterial infection and premature placental separation, was 10.88 (95 % confidence interval: 2.15–55.16). EHV-1 was the main viral cause of pregnancy loss in this study, also associated with other etiological agents, including EHV-2 and EHV-5. The latter viruses in particular need to be more fully investigated to elucidate what role either or both may play as co-infecting agents with other established infectious causes of reproductive disease.     

The new French 2010 Rabbit Hemorrhagic Disease Virus causes an RHD-like disease in the Sardinian Cape hare (Lepus capensis mediterraneus)

Lagovirus is an emerging genus of Caliciviridae, which includes the Rabbit Hemorrhagic Disease Virus (RHDV) of rabbits and the European brown hare syndrome virus (EBHSV) of hares that cause lethal hepatitis. In 2010, a new RHDV related virus (RHDV2) with a unique genetic and antigenic profile and lower virulence was identified in France in rabbits. Here we report the identification of RHDV2 as the cause in Sardinia of several outbreaks of acute hepatitis in rabbits and Cape hare (Lepus capensis mediterraneus). This is the first account of a lagovirus that causes fatal hepatitis in both rabbits and hares.     

Effects of municipal solid waste compost amendments on soil enzyme activities and bacterial genetic diversity

Municipal solid waste (MSW) composts have been used to maintain the long-term productivity of agroecosystems and to protect the soil environment from overcropping, changes in climatic conditions and inadequate management; they also have the additional benefit of reducing waste disposal costs. Since MSW may contain heavy metals and other toxic compounds, amendments cannot only influence soil fertility, but may also affect the composition and activity of soil microorganisms. The effects of MSW compost and mineral N amendments in a 6-year field trial on some physical-chemical properties, enzyme activities and bacterial genetic diversity of cropped plots (Beta vulgaris-Triticum turgidum rotation) and uncropped plots were investigated. The compost was added at the recommended and twice the recommended dosage (12, 24 t ha⁻¹). Amendments of cropped plots with MSW compost increased the contents of organic C from 13.3 to 15.0 g kg⁻¹ soil and total N from 1.55 to 1.65 g kg⁻¹ soil. There were significant increases in dehydrogenase (9.6%), β-glucosidase (13.5%), urease (15.4%), nitrate reductase (21.4%) and phosphatase (9.7%) activities. A significant reduction in protease activity (from 3.6 to 2.8 U g⁻¹ soil) was measured when a double dose of compost was added to the cropped plots. No dosage effect was detected for the other enzymes. Changes in the microbial community, as a consequence of MSW amendment, were minimal as determined using denaturing gradient gel electrophoresis, rDNA internal spacer analysis and amplified ribosomal DNA restriction analysis of bacteria, archaea, actinomycetes, and ammonia oxidizers. This indicates that there was no significant variation in the overall bacterial communities nor in selected taxonomic groups deemed to be essential for soil fertility.     

Synthesis and Antiproliferative Activity of Thiazolyl-bis-pyrrolo[2,3-b]pyridines and Indolyl-thiazolyl-pyrrolo[2,3-c]pyridines,Nortopsentin Analogues

Two new series of nortopsentin analogues, in which the imidazole ring of the natural product was replaced by thiazole and indole units were both substituted by 7-azaindole moieties or one indole unit was replaced by a 6-azaindole portion, were efficiently synthesized. Compounds belonging to both series inhibited the growth of HCT-116 colorectal cancer cells at low micromolar concentrations, whereas they did not affect the viability of normal-like intestinal cells. A compound of the former series induced apoptosis, evident as externalization of plasma membrane phosphatidylserine (PS), and changes of mitochondrial trans-membrane potential, while blocking the cell cycle in G2/M phase. In contrast, a derivative of the latter series elicited distinct responses in accordance with the dose. Thus, low concentrations (GI₃₀) induced morphological changes characteristic of autophagic death with massive formation of cytoplasmic acid vacuoles without apparent loss of nuclear material, and with arrest of cell cycle at the G1 phase, whereas higher concentrations (GI₇₀) induced apoptosis with arrest of cell cycle at the G1 phase.     

Role of secondary metabolites in the biocontrol activity of <Emphasis Type="Italic">Pseudomonas corrugata</Emphasis> and <Emphasis Type="Italic">Pseudomonas mediterranea</Emphasis>

Colletotrichum gloeosporioides is the causal agent of Camellia oleifera anthracnose, mainly infecting fruits and leaves. The fungus secretes degrading enzymes to destroy the cuticle of aerial plant parts and help infect the host successfully. To validate whether a cutinase gene (CglCUT1) was required for cutinase activity and pathogenicity of C. gloeosporioides, the CglCUT1 gene was cloned and analyzed. The characterization of CglCUT1 predicted protein suggests that the cloned DNA encoded a cutinase in C. gloeosporioides affecting C. oleifera. The CglCUT1 showed a high homology to those from C. gloeosporioides causing papaya anthracnose and C. capsici causing pepper anthracnose, as well as those of other ascomycetes. The whole CglCUT1 gene was knocked-out and the knockout mutant (?CglCUT39) was subsequently complemented using Agrobacterium tumefaciens mediated transformation. The knockout transformants exhibited significant decreases in cutinase activity and virulence compared with the wild-type strain. The complemented transformants of the disrupted transformant ?CglCUT39 showed a significant increase in cutinase activity and virulence compared with the disrupted transformant ?CglCUT39. This study suggests that the CglCUT1 gene has a positive effect on fungal virulence of the hemibiotrophic C. gloeosporioides on C. oleifera.     

Ecological thresholds in herb communities for the management of suburban fragmented forests

Fragmentation is a major cause of biodiversity loss. The relationship between fragmentation and biodiversity might be non-linear, due to the presence of ecological thresholds; however, the application of the threshold concept on the conservation of herb biodiversity has received limited attention. We assessed the relationship between a pool of woodland indicators and features of forest fragments (area, isolation, shape), and we evaluated whether ecological thresholds might provide guidelines for conservation. We performed our study in a human dominated area of northern Italy. In order to obtain a group of floristic indicators, we chose perennial herbs characteristic of the phytosociological syntaxa Fagetalia sylvaticae, Carpinion betuli and Erythronio-Carpinion, with suitable Landolt L and H index, obtained from previous relevés in reference and high-quality forests. Subsequently, we analyzed the relationship between richness of indicators and patch features recorded in a highly fragmented landscape. Generalized linear models showed that the number of forest indicators was positively related to fragment size and connectivity. Generalized Additive Models showed that the relationship between indicator richness and fragment size was non-linear, with the presence of two thresholds. The number of forest indicators increased until 35–40 ha; above this size the richness remained constant. Fragments above this threshold should be the target of conservation. Furthermore, the relationship between area and number of indicators was less steep in patches smaller than 1–1.5 ha. Patches between these two thresholds can be optimal targets of management actions, as small size increases can greatly improve their value for biodiversity. The identification of two thresholds in the relationship between patch area and indicators suggests an abrupt response to changing landscape features and provide quantitative targets for conservation.     

Autophagy-dependent anticancer immune responses induced by chemotherapeutic agents in mice

Antineoplastic chemotherapies are particularly efficient when they elicit immunogenic cell death, thus provoking an anticancer immune response. Here we demonstrate that autophagy, which is often disabled in cancer, is dispensable for chemotherapy-induced cell death but required for its immunogenicity. In response to chemotherapy, autophagy-competent, but not autophagy-deficient, cancers attracted dendritic cells and T lymphocytes into the tumor bed. Suppression of autophagy inhibited the release of adenosine triphosphate (ATP) from dying tumor cells. Conversely, inhibition of extracellular ATP-degrading enzymes increased pericellular ATP in autophagy-deficient tumors, reestablished the recruitment of immune cells, and restored chemotherapeutic responses but only in immunocompetent hosts. Thus, autophagy is essential for the immunogenic release of ATP from dying cells, and increased extracellular ATP concentrations improve the efficacy of antineoplastic chemotherapies when autophagy is disabled.