Chemical, antioxidant, and antifungal analysis of oregano and thyme essential oils from Ecuador: Effect of thyme against Lasiodiplodia theobromae and its application in banana rot.

The objective of this study was to evaluate the antioxidant capacity by spectrophotometric methods, the in vitro and in vivo antifungal effect against Lasiodiplodia theobromae and the constitution of the essential oils (EO) of oregano and thyme in comparison with their commercial counterparts. The results showed by the EOs of extracted thyme (T-EO), commercial thyme (CT-EO), extracted oregano (O-EO) and commercial oregano (CO-EO), demonstrated antioxidant profiles with a radical neutralizing potential (DPPH•) of IC50: 1.11 ± 0.019; 1.08 ± 0.05; 40.56 ± 0.227 and 0.69 ± 0.004 mg/mL, respectively. They also revealed a ferric ion reducing capacity (FRAP) of 93.05 ± 0.52; 97.72 ± 0.42; 21.85 ± 0.57 and 117.24 ± 0.64 mg Eq Trolox/g. A reduction in ß-carotene degradation of 65.71 ± 0.04; 51.97 ± 0.66; 43.58 ± 1.56 and 57.46 ± 1.56 %. A total phenol content (Folin-Ciocalteu) of 132.97 ± 0.77; 141.89 ± 2.56; 152.04 ± 0.10 and 25.66 ± 0.40 mg EGA/g. Chemical characterization performed by gas chromatography mass spectrometry (GC-MS) showed that the respective major components of the samples were thymol (T-EO: 45.78 %), thymol (CT-EO: 43.57 %), alloaromadendrene (O-EO: 25.17 %) and carvacrol (CO-EO: 62.06 %). Regarding antifungal activity, it was evident that at the in vitro level, both commercial EOs had a MIC of 250 ppm while the extracted thyme EO had a MIC of 500 ppm; In vivo studies demonstrated that the application of thyme EO had a behavior similar to the synthetic fungicide, slowing down rot in bananas under storage conditions. Finally, partial least squares discriminant analysis (PLS-DA) and heat maps suggest p-cymene, carvacrol, linalool, eucalyptol, 4-terpineol, (z)-ß-terpineol, alkanhol, caryophyllene, ß-myrcene, d-limonene, α-terpinene, α-terpineol, d-α-pinene, camphene, caryophyllene oxide, δ-cadinene, terpinolene and thymol as relevant biomarkers associated with the assessed bioactive properties demonstrating the potential of extracted essential oils for the development of a botanical biofungicide.

Validation of a method of broth microdilution for the determination of antibacterial activity of essential oils.

OBJECTIVE: The aim of the present study was to adapt and optimize a broth microdilution method and compare it to the agar dilution method for the evaluation of activity of essential oils from medicinal plants against Gram-negative bacteria. Based on bibliographic research, active and not active oils were selected. The sensitivity and specificity were established as parameters for validation. The comparison between both methods was made using contingency analysis tables, based on the observed frequencies. For both methods, the minimum inhibitory concentration was determined against Escherichia coli strains, in an essential oil concentration range between 0.03 and 0.48% (v/v). RESULTS: A stable emulsion formation was achieved with the addition of Tween 80 and constant agitation, guaranteeing the continuous contact of oil with bacteria (critical step in the microdilution method). The statistical analysis of results obtained with both methods presented a good sensitivity and specificity (100% in both cases), which let us correctly discriminate between active and non-active oils. The values obtained for the minimal inhibitory concentration were independent of the technique used. Finally, the obtained results show that the validated microtechnique allows important diminishment of time and resources for investigations dealing with essential oils or lipophilic extracts evaluation.

Plants and Natural Products with Activity against Various Types of Coronaviruses: A Review with Focus on SARS-CoV-2.

COVID-19 is a pandemic disease caused by the SARS-CoV-2 virus, which is potentially fatal for vulnerable individuals. Disease management represents a challenge for many countries, given the shortage of medicines and hospital resources. The objective of this work was to review the medicinal plants, foods and natural products showing scientific evidence for host protection against various types of coronaviruses, with a focus on SARS-CoV-2. Natural products that mitigate the symptoms caused by various coronaviruses are also presented. Particular attention was placed on natural products that stabilize the Renin-Angiotensin-Aldosterone System (RAAS), which has been associated with the entry of the SARS-CoV-2 into human cells.

Anti-inflammatory and sedative activities of Peperomia galioides: in vivo studies in mice.

Aerial parts (leaves, flowers, stem) of Peperomia galioides extract administered to mice, was used to confirm its anti-inflammatory and sedative folk uses. The anti-inflammatory activity was assessed by croton oil-induced ear oedema and myeloperoxidase (acute inflammation); cotton pellet-induced granuloma (sub-acute inflammation) and Escherichia coli Lipopolysaccharide (LPS) induced inflammation (cellular mediators). The sedative activity was studied by the pentobarbital-induced sleeping time test. Single doses (300 and 600 mg/kg; i.p.) of the extract reduced croton oil-induced ear oedema and myeloperoxidase activity. Six days administration of the extract (300 mg/kg, i.p.) to mice implanted with cotton pellets diminished granuloma formation. LPS (20 mg/kg, i.p.) enhanced plasma nitrites and TNF-α levels that were inhibited by the extract. The duration but not the onset of sleeping time was enhanced by 300 and 600 mg/kg of the extract. Our results show that P. galioides has anti-inflammatory and sedative activities in mice, which validates its traditional use.

Changes in phytochemical composition, bioactivity and in vitro digestibility of guayusa leaves (Ilex guayusa Loes.) in different ripening stages.

BACKGROUND: Guayusa (Ilex guayusa Loes.) leaves, native of the Ecuadorian Amazon, are popularly used for preparing teas. This study aimed to assess the influence of leaf age on the phenolic compounds and carotenoids and the bioactivity and digestibility (in vitro) of aqueous and hydroalcoholic leaf extracts. RESULTS: In total, 14 phenolic compounds were identified and quantified. Chlorogenic acid and quercetin-3-O-hexose were the main representatives of the hydroxycinnamic acids and flavonols respectively. Seven carotenoids were quantified, lutein being the main compound. Ripening affected phenolic content significantly, but there was no significant difference in carotenoid content. Antioxidant capacity, measured by the DPPH• method, was also significantly affected by leaf age. The measurement of in vitro digestibility showed a decrease in phenolic content (59%) as well as antioxidant capacity, measured by the ABTS•+ method, in comparison with initial conditions of the guayusa infusion. Antibacterial and anti-inflammatory activities were assayed with young leaves owing to their higher phenolic contents. Guayusa did not show any antibacterial activity against Escherichia coli ATCC 25922 or Staphylococcus aureus ATCC 25923. Finally, the hydroalcoholic and aqueous extracts exhibited high in vitro anti-inflammatory activity (>65%). CONCLUSION: Young guayusa leaves have potential applications as a functional ingredient in food and pharmaceutical industries. © 2017 Society of Chemical Industry.

Evaluation of anti-inflammatory activity of the methanolic extract from Jungia rugosa leaves in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Jungia rugosa Less (Asteraceae), popularly known in Ecuador as "Carne humana" or "Fompo", is a vine present into the Andean region. It is traditionally used as medicine for the treatment of bruises, cuts and other external inflammatory processes. This study was designed to investigate the anti-inflammatory activity of J. rugosa leaves extract (JRLE) in rodents. MATERIAL AND METHODS: The acute anti-inflammatory activity was evaluated by animal models, including croton oil-induced ear oedema in mice, carrageenan-induced paw oedema in rats and myeloperoxidase (MPO); the chronic anti-inflammatory activity was evaluated by cotton pellet-induced granuloma. RESULTS: Intraperitoneal administration of JRLE (125, 250, 500mg/kg) significantly (p<0.01-0.001) inhibited the croton oil-induced ear oedema and MPO activity in mice; the carrageenan-induced paw oedema in rats was significantly (p<0.05) reduced by 500mg/kg. Repeated (6 days) administration of the extract to mice previously implanted with cotton pellets reduced the formed granuloma (125mg/kg: 11.7%; 250mg/kg: 17.9%; 500mg/kg: 32.4%) but only the inhibition by 500mg/kg reached statistical significance (p<0.01). CONCLUSIONS: The results show that JRLE is effective as an anti-inflammatory agent in acute and chronic inflammation in mice, supporting its traditional use.

Anticonvulsant activity of bisabolene sesquiterpenoids of Curcuma longa in zebrafish and mouse seizure models.

Turmeric, obtained from the rhizomes of Curcuma longa, is used in South Asia as a traditional medicine for the treatment of epilepsy. To date, in vivo studies on the anticonvulsant activity of turmeric have focused on its principal curcuminoid, curcumin. However, poor absorption and rapid metabolism have limited the therapeutic application of curcumin in humans. To explore the therapeutic potential of turmeric for epilepsy further, we analyzed its anticonvulsant activity in a larval zebrafish seizure assay. Initial experiments revealed that the anticonvulsant activity of turmeric in zebrafish larvae cannot be explained solely by the effects of curcumin. Zebrafish bioassay-guided fractionation of turmeric identified bisabolene sesquiterpenoids as additional anticonvulsants that inhibit PTZ-induced seizures in both zebrafish and mice. Here, we present the first report of the anticonvulsant properties of bisabolene sesquiterpenoids and provide evidence which warrants further investigation toward the mechanistic understanding of their neuromodulatory activity.

Biodistribution and pharmacokinetics of PEG-10kDa-cholecystokinin-10 in rats after different routes of administration.

Cholecystokinin, produced in the proximal small intestine, is a short acting satiating peptide hormone. CCK-10, before and after mono-iodination, was previously coupled to 10kDa polyethylene glycol (PEG). The formed conjugates PEG10kDa-CCK-10 and PEG10kDa-[(127)I]-CCK-10 show after i.p. administration to rats a sustained food intake reduction during 8h in comparison to 2h for free CCK-10. The present study examined the blood pharmacokinetics of this pharmacological interesting molecule by means of PEG10kDa-[(123)I]-CCK-10 following intravenous, intraperitoneal, intramuscular and nasal administration and the biodistribution after i.p. administration. HPLC analysis with radiometric detection allowed the differentiation between inorganic iodide and the intact tracer in blood. Blood kinetics after i.v. injection was fitted to a bi-exponential with a distribution half-life of 15 min and with an elimination half-life of 8 hours for intact PEG10kDa-[(123)I]-CCK-10. The biodistribution studies showed a higher accumulation of the tracer for all administration routes in organs expressing CCK receptors localized in the gastrointestinal tract such as pancreas, duodenum and small intestine. No indication of blood brain barrier crossing for the conjugate could be observed independently of the administration route. Main clearance was via the urinary pathway.

Lack of tolerance development with long-term administration of PEGylated cholecystokinin.

Cholecystokinin (CCK) is a short acting satiating peptide hormone produced in the proximal small intestine. Daily CCK injection in rats initially inhibits food intake, but after several days, food intake is no longer affected, suggesting development of tolerance. Previously, we covalently coupled CCK to a 10kDa polyethylene glycol (mPEG-OH) and showed that this conjugate, PEG-CCK(9), produced a significantly longer anorectic effect than unmodified CCK(9). The present study examined whether tolerance to the anorectic effect develops during long-term administration of PEG-CCK(9). For 14 consecutive days, male Wistar rats (n=12) received a daily i.p injection of 8microgkg(-1) of PEG-CCK(9) and a control group received a daily control injection of mPEG-OH. Body weight and food intake were monitored daily during the experiment. Effects on the pancreas were investigated. On each day, injection of PEG-CCK(9) induced an anorectic effect lasting 3-6h, but failed to significantly reduce daily total food intake compared to controls. The body weight gain of the PEG-CCK(9)-treated animals was not different from controls. The PEG-CCK(9)-treated group had a significantly higher pancreas weight, mainly due to hyperplasia. In conclusion, PEG-CCK(9) continued to have a daily suppressive effect on food intake when administered for 14 consecutive days, showing there was no development of tolerance.

PEGylation of cholecystokinin prolongs its anorectic effect in rats.

The anorectic compound CCK-9 was coupled to polyethylene glycol 5 kDa, 10 kDa, 20 kDa and 30 kDa, under different reaction conditions. Conjugates were purified by HPLC and characterized by MALDI-TOF MS. A 96% PEGylation yield was obtained in buffer pH 7.5 after 6h reaction at 20 degrees C. The anorectic activity was tested in vivo in rats. A single bolus intra-peritoneal injection of non-modified CCK-9 resulted in a significant initial food intake reduction 30 min after food presentation (87% compared to paired control group). When PEG-CCK-9 conjugates modified with polymers of molecular weight up to 20 kDa were injected, lower but statistically significant initial food intake reductions were obtained (76% for PEG 10 kDa-CCK-9 conjugate compared to control group). The cumulative food intake reduction of non-modified CCK-9 is normalized within 1-2h, whereas the PEG-CCK-9 molecules showed a prolonged anorectic activity lasting for 6h for PEG 5 kDa-CCK-9; 23 h for PEG 10 kDa-CCK-9 and between 8h and 23 h for PEG 20 kDa-CCK-9. For PEG 30 kDa-CCK-9 conjugate, neither an initial nor a cumulative FI reduction was observed. PEG-CCK-9 conjugates show a significantly prolonged anorectic activity in comparison to the non-modified peptide. This effect is most evident for the PEG 10 kDa-CCK-9 conjugate.