Phytocomplexes from liquorice (Glycyrrhiza glabra L.) leaves--chemical characterization and evaluation of their antioxidant, anti-genotoxic and anti-inflammatory activity

Three extracts of different polarities of Glycyrrhiza glabra L. leaves were characterized and evaluated for their antioxidant, anti-genotoxic and anti-inflammatory activity. In total, thirty components have been identified and quantified through the use of liquid chromatography (LC) with ultraviolet-visible diode-array-detector (UV-vis-DAD) and mass spectrometry (MS). The main components belong to the polyphenols family, being flavonoid and dihydrostilbene derivatives. The extracts have been investigated for their antioxidant, anti-genotoxic and anti-inflammatory activities, which are fundamental requirements of efficacious chemo-preventive agents. The ethyl acetate extract proved to be the most valuable, evidently for the conspicuous presence of several polyphenols, namely flavonoids and dihydrostilbenes.     

Environmental factors related to damage by Heterobasidion abietinum in Abies alba forests in Southern Italy

A study was carried out in southern Italy to determine the role of some factors in favouring Heterobasidion annosum s. l. spread and damage in eight Silver-fir (Abies alba Mill.) forests. All the studied stands were colonised by H. abietinum. The presence of the fungus (as carpophores on colonised wood) was detected by means of transects and varied from 7.0 to 56.0% of trees. Stand history, precipitation and forest origin, analysed by the χ²-test, had a significant effect on rot severity. Damage was more severe in stands more xerophytic where the woods were planted on pasturelands.     

Interaction of four monoterpenes contained in essential oils with model membranes: implications for their antibacterial activity

The present article reports the antimicrobial efficacy of four monoterpenes (thymol, carvacrol, p-cymene, and gamma-terpinene) against the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Escherichia coli. For a better understanding of their mechanism of action, the damage caused by these four monoterpenes on biomembranes was evaluated by monitoring the release, following exposure to the compounds under study, of the water-soluble fluorescent marker carboxyfluorescein (CF) from large unilamellar vesicles (LUVs) with different lipidic composition (phosphatidylcholine, PC, phosphatidylcholine/phosphatidylserine, PC/PS, 9:1; phosphatidylcholine/stearylamine, PC/SA, 9:1). Furthermore, the interaction of these terpenes with dimyristoylphosphatidylcholine multilamellar vesicles as model membranes was monitored by means of differential scanning calorimetry (DSC) technique. Finally, the results were related also with the relative lipophilicity and water solubility of the compounds examined. We observed that thymol is considerably more toxic against S. aureus than the other three terpenes, while carvacrol and p-cymene are the most inhibitory against E. coli. Thymol and carvacrol, but not gamma-terpinene and p-cymene, caused a concentration-dependent CF leakage from all kinds of LUVs employed; in particular, thymol was more effective on PC and PC/SA LUVS than on PC/PS vesicles, while carvacrol challenge evoked a CF leakage from PC/PS LUVs similar to that induced from PC/SA LUVs, and lower than that measured with PC vesicles. Concerning DSC experiments, these four terpenes caused a decrease in Tm and (especially carvacrol and p-cymene) DeltaH values, very likely acting as substitutional impurities. Taken together, our findings lead us to speculate that the antimicrobial effect of thymol, carvacrol, p-cymene, and gamma-terpinene may result, partially at least, from a gross perturbation of the lipidic fraction of the plasmic membrane of the microorganism. In addition to being related to the physicochemical characteristics of the compounds (such as lipophilicity and water solubility), this effect seems to be dependent on the lipidic composition and net surface charge of the microbic membranes. Furthermore, the compounds might cross the cell membranes, thus penetrating into the interior of the cell and interacting with intracellular sites critical for antibacterial activity.     

Positive correlation between high levels of ochratoxin A and resveratrol-related compounds in red wines

The natural occurrence of ochratoxin A in red wines has been widely reported by several authors, as well as a that of group of stilbenes including cis- and trans-resveratrols and related glucosylated forms. In the present study 112 samples of retail red wines were collected from northern (17), central (46), and southern (49) Italy and were analyzed for both ochratoxin A and resveratrol-related stilbenes. The mean levels of total resveratrols and total piceids were 3.14 and 5.80 mg/L, respectively, whereas the ochratoxin A mean level was 0.64 microg/L. The Merlot wines showed the highest mean value of total stilbenes, followed by Negroamaro and Negroamaro blend, Aglianico, and Syrah, all with mean levels of >10 mg/L. Ochratoxin A was detected in 70, 59, and 100% of wine samples from northern, central, and southern Italy, with mean levels of 0.12, 0.07, and 1.36 microg/L, respectively. The highest values of ochratoxin A were recorded in Negroamaro- and Primitivo-based wine samples from southern Italy, showing also the highest content of stilbenes. In wine samples from southern Italy, a positive correlation was obtained between levels of ochratoxin A and total stilbenes (r = 0.74) as well as between ochratoxin A and total resveratrols (r = 0.50) and between ochratoxin A and total piceids (r = 0.74). These results suggest that toxic levels of ochratoxin A in red wine may be, at some extent, counterbalanced by the beneficial effects of resveratrol derivatives. Further investigation should be warranted in this regard.     

Detection of Leishmania infantum DNA by real-time PCR in canine oral and conjunctival swabs and comparison with other diagnostic techniques

The use of non invasive sampling, such as collection of conjunctival swabs, as a diagnostic tool for the detection of Leishmania DNA is of interest. The purpose of this study was to evaluate the diagnostic utility of detecting Leishmania infection with the use of conjunctival swab samples in dogs living in a highly endemic area for leishmaniosis and to investigate, for the first time, the presence of Leishmania DNA in oral swabs in the same population. One hundred sixty-three dogs living outdoor and recruited in various provinces of Sicily were studied. Leishmania infantum indirect fluorescent antibody test (IFAT), delayed-type hypersensitivity reaction to leishmanin (DTH) and real-time PCR of blood (BL), lymph node (LN), conjunctival (CS) and oral swab (OS) samples were performed. The positive PCR percentages in LN, CS, OS and BL samples were: 24.5%, 22.1%, 8.7% and 5.5%, respectively. Serological and DTH positive percentages were 27.0% and 73.8%, respectively. Seropositive and LN-PCR positive dogs had a high likelihood to be positive by CS-PCR. The similar positive PCR percentages found in CS and LN samples suggest the use of CS-PCR as non-invasive alternative technique to LN-PCR for the detection of Leishmania infection in dogs. In addition, this study demonstrated, for the first time, the presence of Leishmania DNA in oral swabs in dogs.     

Evaluation of the presence of selected viral and bacterial nucleic acids in pericardial samples from dogs with or without idiopathic pericardial effusion

Many viruses have been identified in pericardial fluid and in tissue samples from humans with pericarditis by means of molecular diagnostics. In canine idiopathic pericardial effusion there is as yet no conclusive evidence to support the involvement of an infectious agent. This study was designed to investigate a possible relationship between idiopathic pericardial effusion in dogs and viruses most commonly encountered in humans affected with viral pericarditis. Coxsackievirus B3 RNA, influenza virus type A RNA, human adenovirus type 2 DNA, human cytomegalovirus DNA, and parvovirus B19 DNA were investigated using PCR on pericardial effusion samples and pericardial tissue specimens collected from 14 dogs with idiopathic pericardial effusion. PCR was also used to test for two bacteria, Borrelia burgdorferi and Chlamydia pneumoniae. The same microorganisms were also looked for in pericardial effusions or pericardial washes from 10 dogs with neoplastic pericardial effusion, and in samples collected from 10 dogs which died of a non-cardiac disease. One pericardial effusion sample from a dog with the idiopathic form of the disease tested positive for influenza virus type A and sequencing of the amplicon confirmed the PCR result. In another dog from the same group a cytomegalovirus was detected by PCR in the effusion, but sequencing showed this to be a false-positive result. The genomes of the microorganisms investigated were not detected in neoplastic effusions or pericardial washes. The results indicate that viral and bacterial DNA/RNA of relevance for human pericarditis is rare in pericardial samples from dogs with idiopathic pericardial effusion. The finding of influenza type A viral RNA in pericardial fluid from one dog with the idiopathic form of the disease warrants further investigation.     

Molecular basis of the interaction between proteins of plant origin and proanthocyanidins in a model wine system

Plant proteins are being used as a replacement for animal proteins in wine fining. The surface hydrophobicity of plant proteins in four commercial preparations differing for their origin and processing was assessed by using a fluorescent hydrophobic probe in wine-like media. Displacement of the probe by addition of wine phenolics was measured as a way to compare and predict to some extent the efficiency of these proteins in wine fining. It was found that the binding of polyphenols was much more specific than that of the hydrophobic probe. Further analysis of the polyphenol pattern in protein-treated wine-like solutions pointed out two relevant facts: (1) proteins may interfere with the chemistry of the interactions between polyphenols and other wine components; (2) individual protein preparation having different surface hydrophobicities also have different specificities in binding different polymeric forms of the polyphenols and in their substitution products. These findings are related to the possible carry-over of transition metals and may be worth exploring for custom tailoring the fining process. Whether the practical application of the latter finding will call for production and/or screening of plant-derived proteins with features appropriate to this task remains to be investigated. However, the approaches presented in this study may be used for large-scale screening of protein suitability for fining application under laboratory conditions, providing guidelines for their use in actual winemaking applications.     

Immunomodulatory and Antitumoral Activity of Gold Nanoparticles Synthesized by Red Algae Aqueous Extracts

This study reports on the green and cost-efficient synthesis of gold nanoparticles from three different red algae extracts. The nanoparticles synthesized were fully characterized by UV-Vis spectroscopy, HRTEM, and Z-potential. Relevant components occurring in the extracts, such as polysaccharides or phenolic content, were assessed by analytical techniques such as spectrophotometric assays and liquid chromatography. Finally, the antioxidant, antitumoral, and anti-inflammatory potential of both the extracts and the gold nanoparticles synthesized were analyzed in order to determine a possible synergistic effect on the nanoparticles. The results obtained confirmed the obtainment of gold nanoparticles with significant potential as immunotherapeutic agents. The therapeutic potential of these nanoparticles could be higher than that of inert gold nanoparticles loaded with bioactive molecules since the former would allow for higher accumulation into the targeted tissue.     

Analgesic and motor-blocking action of epidurally administered levobupivacaine or bupivacaine in the conscious dog

Objective  To compare the analgesic and motor-blocking effects of epidurally administered levobupivacaine and bupivacaine in the conscious dog.
Study design  Prospective, randomized, cross-over study.
Animals  Six adult female Beagle dogs.
Methods  Each animal received three doses of levobupivacaine or bupivacaine (0.5, 1.0 and 1.5 mg kg⁻¹; concentrations 0.25%, 0.50%, and 0.75%, respectively) in a total volume of 0.2 mL kg⁻¹ by means of a chronically implanted epidural catheter. Onset, duration (through pinch response in the sacral, lumbar and toe areas) and degree of analgesia and motor-blocking status was determined with a scoring system and at regular intervals over 8.5 hours before (baseline) and after drug administration.
Results  Epidurally administered levobupivacaine and bupivacaine had a similar dose-dependent analgesic action with no significant differences in onset (range: 5–8 minutes), duration (bupivacaine: 42 ± 28, 135 ± 68 and 265 ± 68 minutes, and levobupivacaine: 28 ± 33, 79 ± 55 and 292 ± 133 minutes; 0.25%, 0.50%, and 0.75%, respectively) or maximum degree of analgesia. However, levobupivacaine tended to produce a shorter duration of motor block than bupivacaine and the difference in the motor to nociceptive blockade times was significant at the highest dose.
Conclusion  Epidural levobupivacaine produced an analgesic action similar to that of bupivacaine.
Clinical relevance  Epidural levobupivacaine is suitable for clinical use in dogs, mostly at the highest dose if a high degree of analgesia is required.     

Scleral rupture in dogs, cats, and horses

OBJECTIVES: The aim of this retrospective study was to summarize the most frequent clinical signs, ultrasonographic, and histological findings accompanying scleral rupture as a result of blunt trauma in dogs, cats, and horses. ANIMALS STUDIED AND PROCEDURES: Thirty small animals and three horses diagnosed with scleral rupture resulting from blunt trauma. B-mode ultrasonography was performed on 20 animals. Histopathology was carried out on 18 enucleated globes. RESULTS: In small animals, 80% presented hyphema, 60% subconjunctival hemorrhage, and 53% eyelid and conjunctival swelling. In horses, 100% presented eyelid and conjunctival swelling, 67% hyphema, subconjunctival hemorrhage, and collapsed anterior chamber. Ultrasonographic findings were an area with ill-defined scleral margins (90%), echoic/hyperechoic contents in the anterior and posterior chamber (55%) and in the vitreous (80%). In small animals, scleral rupture location noted on gross examination was: at the posterior pole (4), close to the optic nerve (3), near the limbus (2), and in the dorsal aspect of the globe (1). In horses, the lesion was located at the limbus (3). In small animals, histopathology showed presence of hemorrhage in the anterior, posterior chamber, and vitreous (94%), retinal detachment (94%), choroidal edema and hemorrhages (88%), and choroidal detachment as a result of suprachoroidal hemorrhage (88%). The same lesions were found in the globes of two horses. In small animals, rupture location noted on histopathology was: at the posterior pole (8), close to the optic nerve (4), near the limbus (1), near the ciliary body (1). CONCLUSIONS: The most frequent clinical signs observed were hyphema, subconjunctival hemorrhage, and eyelid and conjunctival swelling. Ultrasonographic findings suggestive for scleral rupture were ill-defined scleral borders and/or echoic/hyperechoic material in the cavities of the globe. On histopathology, lesions severely altering the anatomy of the eye structures were: hemorrhage into the chambers of the globe, subretinal and suprachoroidal hemorrhage leading to retinal and choroidal detachment, respectively. In small animals, the most frequent locations for scleral rupture were the posterior pole and close to the optic nerve, whereas in horses it was the limbus.