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.     

Development of specific diagnostic test for small ruminant lentivirus genotype E

Small ruminant lentivirus (SRLV) belonging to the highly divergent genotype E has recently been identified in the Italian goat breed Roccaverano. In this report we have developed a specific serological test based on recombinant matrix/capsid antigen fusion protein. Performance has been evaluated and compared with a similar test based on genotype B antigen. Herds under study were selected according to the infectious status characterized by blood PCR and sequencing. Results clearly showed that B and E based recombinant ELISA only detected homologous infection and an apparent cross-reactivity was recorded in a herd in which co-infection was present. Three commercially available ELISAs showed different abilities in detecting genotype E infection, being the whole virus-based immunoassay the best choice. Genotype E-recombinant antigen was not detected in ELISA by three commercially available Mabs known to be cross-reactive among CAEV and MVV capsid antigens, further supporting the high divergence of the E genotype from others. Finally, a SRLV-free herd according to commercial ELISA testing, was analysed in the same area where genotype E was identified and few animals belonging to Roccaverano breed were found slightly reactive with the E antigens. Our results suggest that the prevalence of genotype E in other small ruminant populations may be conveniently estimated using a comparative assay based on a combination of genotype specific recombinant antigens and may highlight a wider space in which SRLVs evolve.     

Chromium ions toxicity on the membrane transport mechanism in segments of maize seedling roots

Chromium ions affect the proton extrusion and the potassium uptake capacity in segments of maize seedling roots. The hypothesis that these effects can explain the chromium ions toxicity in plants is discussed.     

Distribution,abundance, and movements of the bottlenose dolphin (Tursiops truncatus) in the Pelagos Sanctuary MPA (north‐west Mediterranean Sea)

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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.     

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.     

Corynebacterium urealyticum urinary tract infection in a cat with urethral obstruction

Corynebacterium urealyticum is an uncommon cause of urinary tract infections in cats. However, it is difficult to diagnose and if left untreated it may result in irreversible bladder lesions. C urealyticum is a multiantibiotic-resistant bacterium whose culture requires special care. Risk factors for the occurrence of this infection include urological procedures, foreign bodies, bladder mucosa abnormalities, immuno-suppressed states and antibiotic treatment. This report describes an unusual case of C urealyticum urinary infection in a young cat with pre-existing urethral obstruction. C urealyticum was isolated in pure cultures from two urine samples. Clinical and ultrasound features, results of the urinalysis and urine culture are described as well as therapeutic treatment and eventual favourable outcome to treatment with amoxycillin-clavulanic acid.     

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.     

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.