Inconspicuous post-mortem findings in rabbits from Switzerland naturally -infected with Rabbit Haemorrhagic -Disease Virus 2.

INTRODUCTION: Rabbit Haemorrhagic Disease Virus 2 (RHDV-2) emerged in France in 2010. In Switzerland, RHDV-2 was first identified in 2015 and apparently has almost replaced the classical Rabbit Haemorrhagic Disease Virus (RHDV) by now. Like RHDV, RHDV-2 causes a viral hepatitis with a peracute course and an increased mortality rate within the rabbitry. RHDV infection causes consistent gross pathological findings, especially in the liver and respiratory tract. Reports about gross pathology for animals naturally infected with RHDV-2 is scarce. The present study analysed the anamnesis and necropsy reports of 35 rabbits examined during routine diagnostics between March 2015 and May 2017. A reverse transcriptase real-time polymerase chain reaction (RT-qPCR) specific for RHDV-2 and RHDV proved a total of 25 animals to be positive for RHDV-2, while none was positive for RHDV. Additionally, histological examinations were performed on liver, lung, and kidney of 18 rabbits that had tested positive by RHDV-2 RT-qPCR. The anamnestic report more often stated an increased mortality rate in RHDV-2 positive (16/18, 89 %) compared to RHDV-2 negative rabbits (3/9, 33 %). Gross pathology did not reveal any pathognomonic changes in RHDV-2 positive animals. Histologically, the liver showed the most severe lesions followed by lung and kidney. Animals positive for RHDV-2 frequently showed signs of gastro-intestinal disease (n = 5) and/or septicaemia (n = 6) masking possible indicators of an RHDV-2 infection, such as the rather unspecific findings of an enlarged spleen or an enlarged, friable, tan-coloured liver. The authors want to raise awareness among clinicians and pathologists that in case of sudden death in commercial or pet rabbits, RHDV-2 needs to be considered as differential diagnosis and should be confirmed by laboratory diagnosis.

INTRODUCTION: Le virus 2 de la maladie hémorragique du lapin (RHDV-2) est apparu en France en 2010. En Suisse, le RHDV-2 a été identifié pour la première fois en 2015 et semble avoir presque remplacé le virus classique de la maladie hémorragique du lapin (RHDV). Comme le RHDV, le RHDV-2 provoque une hépatite virale avec une évolution suraiguë et un taux de mortalité élevé chez les lapins. L'infection par le RHDV entraîne des constatations pathologiques bruts cohérents, notamment au niveau du foie et des voies respiratoires. Les rapports sur la pathologie macroscopique des animaux naturellement infectés par le RHDV-2 sont rares. La présente étude a analysé les rapports d'anamnèse et de nécropsie de 35 lapins examinés lors de diagnostics de routine entre mars 2015 et mai 2017. Une réaction en chaîne par polymérase en temps réel à la transcriptase inverse (RT-qPCR) spécifique pour le RHDV-2 et le RHDV a prouvé qu'un total de 25 animaux étaient positifs pour le RHDV-2, tandis qu'aucun n'était positif pour le RHDV. De plus, des examens histologiques ont été réalisés sur le foie, les poumons et les reins de 18 lapins qui avaient été testés positifs par RT-qPCR pour le RHDV-2. Le rapport anamnestique faisait plus souvent état d'un taux de mortalité accru chez les lapins RHDV-2 positifs (16/18, 89 %) que chez les lapins RHDV-2 négatifs (3/9, 33 %). La pathologie macroscopique n'a révélé aucun changement pathognomonique chez les animaux RHDV-2 positifs. Sur le plan histologique, le foie présentait les lésions les plus graves, suivi des poumons et des reins. Les animaux positifs pour le RHDV-2 présentaient fréquemment des signes de maladie gastro-intestinale (n = 5) et/ou de septicémie (n = 6) masquant les indicateurs possibles d'une infection par le RHDV-2, tels que les découvertes plutôt peu spécifiques d'une rate hypertrophiée ou d'un foie hypertrophié, friable et de couleur beige. Les auteurs souhaitent sensibiliser les cliniciens et les pathologistes au fait qu'en cas de mort subite chez des lapins d'élevage ou de compagnie, le RHDV-2 doit être considéré comme un diagnostic différentiel et doit être confirmé par un diagnostic de laboratoire.

Gonadal activity in male and female captive fossas (Cryptoprocta ferox) during the mating season.

The fossa is an endangered, mongoose-like carnivore species endemic to Madagascar with a breeding season (BS) in the southern hemisphere spring. For the present study, faecal samples of captive fossas were collected for over 1 year for five males and four non-pregnant females, and for two pregnant animals during the reproductive period. The goal was to assess gonadal activity using non-invasive hormone measurements of faecal testosterone (T) and gestagen metabolites using assays previously established in our laboratory and further validated in this study. All study animals were housed in northern hemisphere zoos. In males, the seasonal T metabolite profile revealed a peak in spring (March). High performance liquid chromatography (HPLC) analysis was used to characterize the faecal hormone metabolite composition. The highest immunoreactivity was detected in the position of dihydrotestosterone, whereas native T was not detected. In the two pregnant females, gestagen metabolite concentrations increased 4-9 days after the initial matings and remained elevated throughout gestation with concentrations dramatically higher than those of the non-pregnant females during the BS. In these females, gestagen metabolite analyses did not reveal a seasonal pattern similar to the males. The HPLC-analysis revealed that the major proportion of immunoreactivity was associated with an unknown metabolite, whereas native progesterone was undetectable. The seasonal hormone pattern of male fossas gives proof of the reproductive seasonality of this species. The elevated 5alpha-pregnan-3beta-ol-20-one levels in pregnant animals allows for the characterization of pregnancy in the fossa based on analysis of faecal steroid metabolite concentrations.

The in vitro anti-denaturation effects induced by natural products and non-steroidal compounds in heat treated (immunogenic) bovine serum albumin is proposed as a screening assay for the detection of anti-inflammatory compounds, without the use of animals, in the early stages of the drug discovery process.

There are emerging ethical issues with regards to the use of animals in the early stages of drug discovery for anti-inflammatory and degenerative diseases from natural products using the activity-directed isolation pathways when many compounds (eg > 100) are present in the crude extract or fraction and are to be tested The above-mentioned is the main reason for proposing the use of the in vitro anti-denaturation (stabilization) effects of heat treated (immunogenic) bovine serum albumin (BSA) as an assay. Current methods used for detecting and isolating a wide range of anti-inflammatory compounds in the early stages of the drug discovery process utilize a large number of animals. When BSA is heated and is undergoing denaturation, it expresses antigens associated to Type III hypersensitive reaction and which are related to diseases such as serum sickness, glomerulonephritis, rheumatoid arthritis and systemic lupus erythematosus. Thus, the assay that is being proposed should be applicable to the discovery of drugs for treating the above mentioned diseases and others, once the compounds stabilize the denaturation process.

In vitro anti-proliferation/cytotoxic activity of epingaione and its derivatives on the human SH-SY5Y neuroblastoma and TE-671 sarcoma cells.

Epingaione (4-Methyl-1-(5-methyl-2, 3,4,5-tetrahydro-[2,3']bifuranyl-5-yl)-pentan-2-one) was isolated as one of the major lipophilic secondary metabolites from the leaves and stems of Bontia daphnoides L. The compound gave 79.24% and 50.83% anti-proliferation/cytotoxic activity on the human SH-SY5Y neuroblastoma and TE-671 sarcoma cells in vitro at 50 pg/mL, respectively. Epingaione was transformed into eleven derivatives under laboratory conditions using ethanol, some gave greater anti-proliferation/cytotoxic activity on the cancer cell lines tested. One of the derivatives (compound 2) with enhanced cytotoxic activity was elucidated as 5'-Ethoxy-5-methyl-5-(4-methyl-2-oxo-pentyl)-2,3,4,5-tetrahydro-5'H-[2,3']bifuranyl-2'-one. Both epingaione and compound 2 caused an accumulation of arrested or dead SH-SY5Y neuroblastoma in the m-phase of the cell cycle as revealed by the m-phase specific marker KE 67.

Role of transient receptor potential ankyrin 1 receptors in rodent models of meningeal nociception - Experiments in vitro.

BACKGROUND: The TRP channel ankyrin type 1 (TRPA1) is a nonselective cation channel known to be activated by environmental irritants, cold and endogenous mediators of inflammation. Activation of TRPA1 in trigeminal afferents innervating meningeal structures has recently been suggested to be involved in the generation of headaches. METHODS: Two in vitro models of meningeal nociception were employed using the hemisected rodent head preparation, (1) recording of single meningeal afferents and (2) release of calcitonin gene-related peptide (CGRP) from the cranial dura mater. The role of TRPA1 was examined using the TRPA1 agonists acrolein and mustard oil (MO). BCTC, an inhibitor of TRP vanilloid type 1 receptor channels (TRPV1), and the TRPA1 inhibitor HC030031 as well as mice with genetically deleted TRPA1 and TRPV1 proteins, were used to differentiate between effects. RESULTS: Acrolein did not cause discharge activity in meningeal Aδ- or C-fibres but increased the electrical activation threshold. Acrolein was also effective in releasing CGRP from the dura of TRPV1-/- but not of TRPA1-/- mice. MO increased the discharge activity of afferent fibres from rat as well as C57 wild-type and TRPA1-/- but not TRPV1-/- mice. The effect was higher in C57 compared to TRPA1-/- mice. CONCLUSION: Sole TRPA1 receptor channel activation releases CGRP and increases the activation threshold of meningeal afferents but does not generate propagated activity, and so would be capable of causing local effects like vasodilatation but not pain generation. In contrast, combined TRPA1 and TRPV1 activation may be rather pronociceptive supporting headache generation. SIGNIFICANCE: Sole activation of TRPA1 receptor channels increases the activation threshold but does not cause propagated action potentials in meningeal afferents. TRPA1 agonists cause CGRP release from rodent dura mater. Peripheral TRPA1 receptors may have a pronociceptive function in trigeminal nociception only in combination with TRPV1.

From Hospital to Home: Limited Nutritional and Functional Recovery for Older Adults.

BACKGROUND: The post-hospital period may be a vulnerable time for elders recovering from acute illness. Few studies have examined nutrition outcomes of older people at nutrition risk after acute hospitalisation. OBJECTIVES: This study aims to describe a) standard nutrition care received by recently discharged older medical patients, b) change in nutritional and functional status at six weeks post-discharge and c) clinical outcomes at twelve weeks post discharge. DESIGN: Prospective cohort study. SETTING: Two metropolitan teaching hospitals in Brisbane, Australia. PARTICIPANTS: Medical patients aged ≥65 years at risk of malnutrition (Malnutrition Screening Score ≥2) and discharged to independent living in the community. MEASUREMENT: Nutritional status (Mini Nutritional Assessment (MNA), weight, lean body mass), functional status (grip strength, walk speed, activities of daily living) and health-related quality of life assessed on discharge and six weeks post-discharge. Inpatient and post-discharge nutrition intervention was recorded. Death and unplanned admissions were measured at 12 weeks. RESULTS: Of the 42 consented participants, only 14% (n=6) received post-discharge dietitian review and 19% (n=8) received practical nutrition supports at home (meal delivery, shopping assistance) as part of standard care. While there was a small improvement in MNA (18.4±4.0 to 20.1±4.2, p=0.004) and walk speed (0.7±0.3 m/s to 0.9±0.3, p=0.004) at six weeks, there was no difference in mean weight, lean body mass, grip strength or activities of daily living. Five (15%) participants lost ≥5% body weight. By twelve weeks, 17 participants (46%) had at least one unplanned hospital admission and four (10%) had died. CONCLUSIONS: Few patients at nutrition risk received nutrition-focussed care in the post-hospital period, and most did not improve nutritional or functional status at 6 weeks.

Transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of Pseudomonas putida 86.

Resting cells of Pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-Dioxygenating activity was present when the enzymes of quinoline catabolism were induced. Quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and D-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. Succinate-grown resting cells did not oxidize 2-chloroquinoline. Acid-catalyzed decomposition of 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline predominantly yielded 2-chloro-8-hydroxyquinoline. By analogy, accumulation of the putative dead-end metabolite 1H-8-hydroxy-2-oxoquinoline during growth of P. putida 86 on quinoline is suggested to likewise result from dehydration of the 7,8-dihydrodiol of 1H-2-oxoquinoline.

Metabolism of 4-chlorophenol by Azotobacter sp. GP1: structure of the meta cleavage product of 4-chlorocatechol.

A mutant strain of Azotobacter sp. GP1 converted 4-chlorophenol to 4-chlorocatechol under cometabolic conditions. Under the same conditions the wild-type strain accumulated a yellow compound, which by chemical and spectroscopic methods was identified as 5-chloro-2-hydroxy-6-oxohexadienoic acid (5-chloro-2-hydroxy-muconic semialdehyde). The structure of this compound indicates a meta-proximal cleavage of 4-chlorocatechol.

Isolation and identification of N-mercapto-4-formylcarbostyril, an antibiotic produced by Pseudomonas fluorescens.

Pseudomonas fluorescens strain G308 isolated from barley leaves produces a novel antibiotic substance that was purified by preparative TLC and HPLC and identified as N-mercapto-4-formylcarbostyril (Cbs) by LC/DAD, IR, LC-ES(+)/MS, LC-ES(-)/MS, GC-EI/MS, LC-HRES(+)/MS, mass isotope ratios analysis, 1H NMR and 13C NMR analysis. The purified new antibiotic compound is effective against many phytopathogenic fungi in vitro. The compound inhibited at 25 ppm spore germination and germ tube growth of the following fungi; Fusarium oxysporum f. sp. lycopersici, Fusarium culmorum, Cladosporium cucumerinum and Colletotrichum lagenarium. At concentrations up to 125 ppm, the compound did not interfere with release of zoospores from sporangia and germination of encysted zoospores of Phytophthora infestans.

Sesquiterpene lactones in Viguiera eriophora and Viguiera puruana (Heliantheae; Asteraceae).

Extracts of the aerial parts of Viguiera eriophora ssp. eriophora and Viguiera puruana afforded, in addition to known compounds, six new heliangolides and a germacrolide, whose structures were determined by spectral analysis. HPLC analysis and LC NMR experiments revealed the natural occurrence of the compounds in glandular trichomes. The taxonomic relevance of the results is briefly discussed.

Sesquiterpene lactones and a myoinositol from glandular trichomes of Viguiera quinqueremis (Heliantheae; Asteraceae).

The extract of the floral parts of Viguiera quinqueremis afforded, in addition to known compounds, six new sesquiterpene lactones as well as a new myoinositol derivative. All compounds were detected in glandular trichomes which were collected micromechanically from the anther appendages and were analyzed by HPLC. Structure identification was performed by 1H NMR measurements including LC NMR and LC MS experiments.

Scaphopetalone and scaphopetalumate, a lignan and a triterpene ester from Scaphopetalum thonneri.

From the methanol extract of the stem bark of Scaphopetalum thonneri, two new compounds, including one lignan, named scaphopetalone, one new ester of ferulic acid, named scaphopetalumate were isolated together with three known compounds including: two coumarins (scopoletin and scopolin), and one pentacyclic triterpene (oleanolic acid). The structure of the new compounds were elucidated by means of spectroscopic analyses.

Application of high-performance liquid chromatography coupled to nuclear magnetic resonance spectrometry, mass spectrometry and bioassay for the determination of active saponins from Bacopa monniera Wettst.

Reversed-phase high-performance liquid chromatographic separation coupled to (structurally informative) spectroscopic methods like NMR and MS and an efficient bioassay have been used to determine the active compounds from a crude fraction of Bacopa monniera. The fraction containing a mixture of saponins with closely related structures was found to show a significant anthelmintic activity against Caenorhabditis elegans (used as a model test organism for determining anthelmintic activity). The activity was correlated to two dammarane type triterpenoidal saponins containing at least three sugar units. The optimization of separation for 1 mg of the crude sample on column and the sensitivity of on-flow one- and two-dimensional NMR experiments to the high-molecular-mass compounds (M(r) 890-930) has been demonstrated.

Combination of LC-MS and LC-NMR as a Tool for the Structure Determination of Natural Products

Application of both LC-MS and LC-NMR to a partially purified extract of Vernonia fastigiataled to the direct identification of antibacterial sesquiterpene lactones 1-9 without isolation of individual compounds. The rapid structural analysis of both major and minor components of this class of compounds demonstrated the power of structure-guided screening as a complementary method to assay-guided screening.

Aloe vera: a systematic review of its clinical effectiveness.

BACKGROUND: The use of aloe vera is being promoted for a large variety of conditions. Often general practitioners seem to know less than their patients about its alleged benefits. AIM: To define the clinical effectiveness of aloe vera, a popular herbal remedy in the United Kingdom. METHOD: Four independent literature searches were conducted in MEDLINE, EMBASE, Biosis, and the Cochrane Library. Only controlled clinical trials (on any indication) were included. There were no restrictions on the language of publication. All trials were read by both authors and data were extracted in a standardized, pre-defined manner. RESULTS: Ten studies were located. They suggest that oral administration of aloe vera might be a useful adjunct for lowering blood glucose in diabetic patients as well as for reducing blood lipid levels in patients with hyperlipidaemia. Topical application of aloe vera is not an effective preventative for radiation-induced injuries. It might be effective for genital herpes and psoriasis. Whether it promotes wound healing is unclear. There are major caveats associated with all of these statements. CONCLUSION: Even though there are some promising results, clinical effectiveness of oral or topical aloe vera is not sufficiently defined at present.

Feverfew for preventing migraine.

BACKGROUND: Feverfew (Tanacetum parthenium L.) is a popular herbal remedy for migraine. OBJECTIVES: To systematically review the evidence for or against the efficacy of feverfew versus placebo for the prevention of migraine. SEARCH STRATEGY: Electronic literature searches were performed using the databases CISCOM (Research Council for Complementary Medicine, London, UK), MEDLINE, EMBASE, Biosis and the Cochrane Library (each from its inception to April 1998). Manufacturers were contacted and the bibliographies of identified articles checked for further trials. SELECTION CRITERIA: Randomised, placebo-controlled, double-blind trials assessing the efficacy of feverfew for preventing migraine were included. No restrictions regarding the language of publication were imposed. DATA COLLECTION AND ANALYSIS: Data on patients, interventions, methods, outcomes and results were extracted in a pre-defined, standardised manner. Methodological quality was evaluated using the scoring system developed by Jadad and colleagues. Both data extraction and the assessment of methodological quality were performed independently by two reviewers. MAIN RESULTS: Four trials met the inclusion criteria. The majority of these trials suggested beneficial effects of feverfew compared with placebo. However, the trial with the highest methodological quality, which was also among the largest, found no significant difference between feverfew and placebo. REVIEWER'S CONCLUSIONS: The efficacy of feverfew for the prevention of migraine has not been established beyond reasonable doubt.

Escuside, a new coumarin-secoiridoid from Fraxinus ornus bark.

A new coumarin-secoiridoid, named escuside, was isolated from the bark of Fraxinus ornus and its structure determined by spectroscopic methods.

Diterpenoid and limonoids from the stem of Pterorhachis zenkeri.

One new diterpenoid, methyl 3alpha-hydroxy-7-oxo-dehydroabietate (1), two new limonoids, 3alpha-deacetyl-amoorastatin (2) and 9beta-amoorastatin (3), and the known limonoid amoorastatin (4) were isolated from the stem of Pterorhachis zenkeri.