Inula viscosa (L.) Aiton leaves and flower buds: Effect of extraction solvent/technique on their antioxidant ability, antimicrobial properties and phenolic profile.

This study was designed to establish the most effective solvent/technique for extracting antioxidant phytoconstituents from leaves and flower buds of Inula viscosa (L.) Aiton (Asteraceae) grown wild in Morocco. Maceration and hot extraction with methanol or water and Soxhlet ethanol extraction were utilized. The antioxidant potential was evaluated in vitro by DPPH, reducing power, and ferrous ions chelating activity assays. I. viscosa leaf and flower bud extracts displayed the strongest effect in the DPPH test, being the Soxhlet ethanol the most active ones (IC50 = 54.24 ± 0.21 µg/mL and 39.77 ± 0.23 µg/mL); thus, they were selected for further investigations. The antimicrobial efficacy of the Soxhlet ethanol extracts against ATCC and food isolates strains was assayed; the leaf extract showed the best activity, and Candida albicans was the most sensitive strain (MIC = 125 µg/mL). The extracts resulted non-toxic against Artemia salina. Among the phenolics characterised by HPLC-PDA-ESI-MS, hispidulin hexoside, patuletin and spinacetin were identified for the first time.

Antiinflammatory and antioxidant activity of Hypericum rumeliacum Boiss. subsp. apollinis (Boiss. & Heldr.) Robson & Strid methanol extract.

Plants of the genus Hypericum are widely used in folk medicine for the treatment of gastric ailments (including pectic ulcers), burns, swelling, inflammation, anxiety, as well as for bacterial and viral infections. The aerial parts of Hypericum rumeliacum Boiss. subsp. apollinis (Boiss. & Heldr.) Robson & Strid (Guttiferae), a perennial herb growing wild in Greece, were subjected to biological and phytochemical studies. The H. rumeliacum methanol extract showed, on the DPPH test, antioxidant activity (IC(50) = 23.61 microg/mL). The antiinflammatory activity of the extract (70 mg/kg, i.p.), in the carrageenan-induced paw oedema in rat, was studied. In this experimental model an inhibition of paw oedema, significant from the first to third hour, was observed. The histological examination confirmed resolution of the inflammatory process observed in vivo. Through HPLC analysis of the H. rumeliacum methanol extract, the naphthodianthrones hypericin, pseudohypericin and some polyphenol compounds, such as chlorogenic acid, rutin and isoquercitrin, were identified and quantified. The antioxidant activity of H. rumeliacum could be chiefly related to polyphenol compounds that are well known as the main contributors to the free radical-scavenging effects. The antiinflammatory effect of the methanol extract of the aerial parts could be due to the synergic action both of hypericin and polyphenol compounds.

Anti-inflammatory activity of extract and fractions from Nepeta sibthorpii Bentham.

Several species of Nepeta genus are utilized in folk medicine for treatment of contusions, rheumatic pains, fever, cutaneous eruptions. Some species are employed for their anti-inflammatory properties. In this paper, we report the results of phytochemical studies on aerial parts of Nepeta sibthorpii Bentham (Lamiaceae), an endemic plant of Greece. The bioassay-guided fractionation of methanol extract led to the isolation of ursolic acid and polyphenol fraction. By HPLC, we determined some phenolics: chlorogenic acid (0.315 mg/g) and the flavonoids rutin (0.091 mg/g), luteolin-7-O-glucoside (0.387 mg/g) and a luteolin derivative. We assayed the radical scavenging activity of Nepeta sibthorpii methanol extract by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Moreover, we studied the anti-inflammatory activity of Nepeta sibthorpii methanol extract (50 mg/kg, os), ursolic acid and polyphenol fraction (dose corresponding to 50 mg/kg of methanol extract, os) in the carrageenan-induced paw oedema in rat. In this experimental model, we observed a significant inhibition of paw oedema. We suppose that the anti-inflammatory effect of methanol extract could be related to the free radical scavenging activity and that it depends on a synergic action of all the components of the methanol extract, even if ursolic acid can be considered the main responsible for this activity.

Biological effect of Opuntia ficus indica (L.) Mill. (Cactaceae) waste matter. Note I: diuretic activity.

In this work we studied in rat the diuretic activity of Opuntia ficus indica (L.) Mill. (Cactaceae) waste matter. The cladodes, flowers and non commerciable fruits were collected in S. Cono (CT, Sicily) cultivation. Acute and chronic diuretic activity of 15% infusion of cladodes, flowers and fruits were assayed. Natriuresis, kaliuresis and the activity on fructose-induced hyperuricemia was also studied. The results show that O. ficus indica cladode, fruit and flower infusions significantly increase diuresis. This effect is more marked with the fruit infusion and it is particularly significant during the chronic treatment. The fruit infusion shows also antiuric effect. In all experiments cladode, flower and fruit infusions showed a modest but not significant increase in natriuresis and kaliuresis.

Study on the increment of the production of gastric mucus in rats treated with Opuntia ficus indica (L.) Mill. cladodes.

Opuntia ficus indica cladodes are used in traditional medicine of many countries for their cicatrisant activity. The major components of cladodes are carbohydrate-containing polymers, which consist of a mixture of mucilage and pectin. In this paper we studied the cytoprotective effects of cladodes on experimental ethanol-induced ulcer in rat. The O. ficus indica cladodes administration gives rise to cytoprotection phenomena by breaking up the epithelial cells and stimulating an increase in mucus production. When O. ficus indica cladodes are administered as a preventive therapy, keep the gastric mucosa under normal condition by preventing mucus dissolution caused by ethanol and favouring mucus production. An increase of mucus production is also observed during the course of the curative treatment. The treatment with O. ficus indica cladodes provokes an increase in the number of secretory cells. Probably, the gastric fibroblasts are involved in the antiulcer activity.

Effects of Teucrium divaricatum Heldr. ssp. divaricatum decoction on experimental ulcer in rats.

The effects of chronic intragastric administration of the aerial parts decoction of Teucrium divaricatum Heldr. ssp. divaricatum (Labiateae) on experimental acetylsalicylic acid (ASA)-induced ulcer were studied in rats. The ulcer index and histological mucosa regeneration were evaluated. The ulcer index significantly decreased (U.I. 1.7+/-1.5; P<0.05) after treatment with a lyophilized decoction of T. divaricatum (500 mg/kg os), once daily for 7 days, compared to the control (U.I. 11.4+/-2.3). The ultrastructure modifications of gastric mucosa were observed by transmission electronic microscopy (TEM) confirming the antisecretory effect exercised by administration of the T. divaricatum decoction. Treatment with T. divaricatum seems to block up parietal cell acid secretion and to stimulate in the chief cells the formation of zymogen granules.

[Entomological survey of sandflies (Diptera, Psychodidae) in the province of L'Aquila (Abruzzo)]. / Indagine entomologica sui flebotomi (Diptera, Psychodidae) in provincia di L'Aquila (Abruzzo).

A survey on phlebotomine sandflies (Diptera, Psychodidae) was carried out in the province of L'Aquila, Abruzzo region (Italy). Ten collecting sites at altitudes between 700 and 1,070 m a.s.l., produced a total of 324 specimens (81.1% males), 72.86% being Phlebotomus perniciosus, 3.08% P. mascittii, 0.30% P. papatasi and 23.76% Sergentomyia minuta. The collecting sites at 1,070, 950 and 775 m a.s.l. are the highest levels recorded in Italy for P. perniciosus, P. mascittii and P. papatasi, respectively. P. perfiliewi, the proven vector of cutaneous leishmaniasis (CL) in the Abruzzo region (Maroli et al., 1987) was absent. The probable role of P. perniciosus in the transmission of the sporadic CL cases reported from the area is discussed.

Nepeta sibthorpii Bentham (Lamiaceae): micromorphological analysis of leaves and flowers.

Nepeta sibthorpii Bentham (Lamiaceae) is an herbaceous aromatic plant, growing in dry, usually rocky areas of Southern Greece. In this paper, we report the study of micromorphological features of the flowers and leaves of Nepeta sibthorpii Bentham. The analysis by means of scanning electron microscopy has permitted to characterize both the covering trichomatous garment, abundantly present on the calyx and on the leaves, and the glandular hairs, distributed on the adaxial leaf surface and on the outer surfaces of calyx and corolla, showing also the peculiar features of the epidermal cells, all useful diagnostic elements for the identification of this medicinal plant.

Anticonvulsant and sedative effects of some 5-substituted bromopyrazolinic spirobarbiturates.

We studied in mice the neuropharmacological effect of some 5-substituted bromopyrazolinic spirobarbiturates. LD50 and CL50 values were estimated for all the investigated compounds. The effect on potentiation of sodium pentobarbital activity and on generalised tonic-clonic seizures produced by PTZ on mice was studied. All the compounds had the same pharmacological profile, they extended sleeping-time induced by sodium pentobarbital and showed protection against PTZ-induced convulsions, decreasing the death rate.

Phenolic composition and biological activities of Juniperus drupacea Labill. berries from Turkey.

The present study was designed to define the phenolic profile and the biological potential of berries methanol extract of Juniperus drupacea Labill. from Turkey. The total phenolic content (Folin-Ciocalteau assay) was 48.06±0.99mgGAE/g extract. The HPLC-DAD-ESI-MS analysis allowed the determination of the complete phenolic profile of J. drupacea berries. Phenolic acids represented more than 60% of the total phenolics, and tyrosol was the major one (1324±0.64µg/g extract); within the flavonoids amentoflavone was detected as the main constituent (927±0.35µg/g extract). The extract exhibited good antioxidant properties, as determined by different in vitro models: DPPH test (IC(50) 0.38±0.02mg/mL), reducing power (12.63±0.14ASE/mL), Fe(2+) chelating ability (IC(50) 2.26±0.06mg/mL), and TBA test (IC(50) 2.47±1.13µg/mL). Cytotoxicity against Artemia salina was highlighted (LC(50) 489.47±27.8µg/mL), and a significant decrease (p⩽0.05; p⩽0.01) in HepG2 cells viability was observed at the higher concentrations (5-10µg/mL). The extract displayed good antibacterial activity towards Gram-positive bacteria and in particular Staphylococcus aureus was the most susceptible strain (MIC 78.12µg/mL).

Doppler and elastography as complementary diagnostic methods for mammary neoplasms in female cats / Doppler e elastografia como métodos diagnósticos complementares de neoplasias mamárias em felinos

Objetivou-se descrever a utilização das técnicas Doppler e elastografia ARFI (acoustic radiation force impulse) na avaliação de neoplasias mamárias de gatas, métodos ainda não utilizados para tal estudo em fêmeas felinas. Após a avaliação física e específica das neoplasias mamárias de duas gatas, foram realizadas as avaliações ultrassonográfica modo-B (ecogenicidade, ecotextura e margeamento), Doppler colorido (característica do fluxo sanguíneo) e espectral (velocidade sistólica/VS; diastólica/VD; e índice de resistência/IR) e a eslatografia ARFI qualitativa (escala de cores) e quantitativa (velocidade de cisalhamento) dos nódulos mamários, avaliando a aplicabilidade destas técnicas para determinar a malignidade dos tumores mamários. Após mastectomia e histopatologia foram diagnosticados: caso 1: carcinoma tubular; e caso 2: carcinoma mamário cribiforme. As características do modo-B foram similares para ambos os casos (heterogeneidade e margens regulares e não invasivas). Ao Doppler verificou-se presença de neovascularização e valores considerados elevados para os índices vasculares (quando comparado com tecidos benignos de outras espécies, como por exemplo caninos) (caso 1 - VS: 23,1cm/s, VD: 8,5cm/s e IR: 0,63; caso 2 - VS: 47,4 e 24,5cm/s; VD: 16,1 e 6,1cm/s e IR: 0,66 e 0,75); assim como os valores da velocidade de cisalhamento dos tecidos (caso 1: 4,07 m/s; caso 2: 4,54 e 6,85 m/s) na elastografia quantitativa; sendo que na avaliação qualitativa observou-se predominante rigidez e não deformidade tecidual, características que podem indicar a malignidade tecidual. Conclui-se que estas técnicas podem auxiliar no estudo de neoplasias mamárias em felinos, sugerindo a implementação destas técnicas de imagem para estudos de tumores mamários em felinos.

Erucin, a new promising cancer chemopreventive agent from rocket salads, shows anti-proliferative activity on human lung carcinoma A549 cells.

Erucin (ER) is a dietary isothiocyanate present in cruciferous vegetables, such as rocket salads (Erucasativa Mill., Diplotaxis sp.), that has been recently considered a promising cancer chemopreventive phytochemical. Biological activity of ER was investigated on human lung adenocarcinoma A549 cells, analyzing its effects on molecular pathways involved in apoptosis and cell cycle arrest, such as PARP-1 cleavage, p53 and p21 protein expression. Our results show that ER affects the A549 cell proliferation, enhancing significantly p53 and p21 protein expression in a dose-dependent manner (p<0.001). PARP-1 cleavage occurs only after exposure to high concentrations of ER (50 microM), in accordance to previous studies showing similar bioactivity of other isothiocyanates (ITCs). Our study reports for the first time that the induction of p53, p21 and PARP-1 cleavage may participate in the anti-proliferative activity of ER in human lung adenocarcinoma A549 cells. Comparison of data with those obtained with the isothiocyanate sulforaphane (SF), structurally related to ER, underlines the strong relationship between structural analogy of ITCs and their biological activity. The ability of dietary compounds to modulate molecular mechanisms that affect cancer cell proliferation is certainly a key point of the cancer prevention potential by functional foods.

Ursolic acid plays a role in Nepeta sibthorpii Bentham CNS depressing effects.

The sedative, anticonvulsant and analgesic activity of ursolic acid, a terpenoid bioassay-isolated from Nepeta sibthorpii Bentham, was evaluated in mice. The oral administration of ursolic acid (2.3 mg/kg) produced a significant depressant effect on CNS by reducing spontaneous motor activity and the number and lethality of pentylenetetrazol (PTZ)-induced seizures. Two models of nociception, the writhing test and the hot plate test, were also used to examine the analgesic effect of ursolic acid. At a dose of 2.3 mg/kg, ursolic acid caused an inhibition of acetic acid-induced abdominal constriction, but was inactive in the hot plate test. Treatment at a higher dose (20 mg/kg) significantly increased the reaction time in the hot plate test. This effect, reversed by naloxone, evidently involves opioid receptors, but the analgesic activity of ursolic acid may be related also to the antiinflammatory and antioxidant properties of this compound.

Opuntia ficus indica (L.) Mill. mucilages show cytoprotective effect on gastric mucosa in rat.

Opuntia ficus indica cladodes possess a protective action against ethanol-induced ulcer in the rat. The major components of cladodes are carbohydrate polymers, mainly mucilages and pectin. To clarify the cytoprotective effects of cladodes on experimental ethanol-induced ulcer in rat, mucilages and pectin were extracted and were administered instead of cladodes. The above mentioned effects induced by cladodes may be attributed to mucilages, and not significantly to pectin.

Opuntia ficus indica (L.) Mill. fruit juice protects liver from carbon tetrachloride-induced injury.

The protective effects of the juice of Opuntia ficus indica fruit (prickly pear) against carbon tetrachloride (CCl(4))-induced hepatotoxicity were examined in rats. The animals were treated orally with the juice (3 mL/rat) 2 h after administration of the hepatotoxic agent. Preventive effects were studied by giving the juice (3 mL/rat) for 9 consecutive days. On day 9 the rats received the hepatotoxic agent. Morphological and biochemical evaluations were carried out 24, 48 and 72 h after induction of the hepatic damage. Data show that O. ficus indica fruit juice administration exerts protective and curative effects against the CCl(4)-induced degenerative process in rat liver. Histology evaluation revealed a normal hepatic parenchyma at 48 h; the injury was fully restored after 72 h. Moreover, a significant reduction in CCl(4)-induced increase of GOT and GPT plasma levels is evident; these data are in agreement with the functional improvement of hepatocytes. O. ficus indica fruit juice contains many phenol compounds, ascorbic acid, betalains, betacyanins, and a flavonoid fraction, which consists mainly of rutin and isorhamnetin derivatives. Hepatoprotection may be related to the flavonoid fraction of the juice, but other compounds, such as vitamin C and betalains could, synergistically, counteract many degenerative processes by means of their antioxidant activity.

Salvadora persica L.: hypolipidemic activity on experimental hypercholesterolemia in rat.

Stems of Salvadora persica are widely used as tooth cleaning sticks in Arabic countries and decoctions show hypocholesterolemic properties. The effects of prolonged administration of a lyophilized stem decoction of Salvadora persica were evaluated in diet-induced rat hypercholesterolemia. The preparation was administered for 15 and 30 days (by gavage at a dose of 500 mg/kg in an aqueous vehicle in a volume of 0.5 ml/100 g b.w.). Cholesterol, HDL, LDL and triglyceride plasma levels were assayed. The results showed that the S. persica decoction significantly lowered cholesterol and LDL plasma levels in rats, proving to be more active at 30 days of treatment. The systemic administration of Triton results in a rise in plasma cholesterol and triglyceride levels. The results obtained show that S. persica decoction was inactive at 18 hr after treatment, whereas at 27 hr, it was able to reduce cholesterol and LDL plasma levels. In all experiments HDL and triglycerides were unchanged.

Effects of naringin on experimental ulcer in rats.

The effects of chronic intragastric administration of naringin (200 mg/kg) on experimental acetylsalicylic acid (ASA)-induced ulcer were studied in rat. The ulcer index and histological mucosa regeneration were evaluated. The ulcer index significantly decreased after treatment with naringin (200 mg/kg) once daily for seven days. Microscopic observations confirm these results.

Anticonvulsant and sedative effects of Salvadora persica L. stem extracts.

The effect of Salvadora persica L. stem extracts on the potentiation of sodium pentobarbital activity and on generalized tonic-clonic seizure produced by pentylenetetrazol (PTZ) on the rat is reported. The extracts of Salvadora persica L. extended sleeping-time and decreased induction-time induced by sodium pentobarbital; in addition it showed protection against pentylenetetrazol-induced convulsion by increasing the latency period and diminishing the death rate.

Chiral 3,3'-(1,2-ethanediyl)-bis[2-(3,4-dimethoxyphenyl)-4-thiazolidinones] with anti-inflammatory activity. Part 11: evaluation of COX-2 selectivity and modelling.

Anti-inflammatory/analgesic 3,3'-(1,2-ethanediyl)-bis[2-(3,4-dimethoxyphenyl)-4-thiazolidinones] 1, obtained as racemic mixtures (a) and mesoforms (b), have two equivalent stereogenic centres (C-2 and C-2') and exist as RR, SS and RS isomers. The enantioseparation of 1a provided the single enantiomers that displayed different in vitro cyclooxygenase-1/cyclooxygenase-2 selectivity ratios. In particular the dextrorotatory compound is a highly selective COX-2 inhibitor and the levorotatory one is moderately selective. Instead, RS-meso isomer (1b) exhibited similar levels of inhibitory activity on both COX isozymes. The diastereo- and enantioselectivity has been explained by molecular modelling of RR, SS and RS compounds into COX-1 and COX-2 binding sites. Theoretical results indicated SS>RS>RR affinity order towards COX-2 isoenzyme, in agreement with in vitro and previous in vivo pharmacological results.