Antimicrobial,modulatory and chemical analysis of the oil of Croton limae

Context: Croton sp. are plants with a well-reported antimicrobial activity. Croton limae A.P. Gomes, M.F. Sales P.E. Berry (Euphorbiaceae), known as ‘marmeleiro-prateado’, is commonly used to manage abdominal pain in Brazil.

Objective: This work evaluates the phytochemical composition, antimicrobial and modulatory activities of the essential oil of C. limae leaves (EOCL).

Materials and methods: The minimum inhibitory concentration (MIC) and the modulation of the antibiotic activity were determined using a microdilution method. The concentration of EOCL ranged between 512 and 8?μg/mL. Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Candida tropicalis, C. krusei and C. albicans strains were used in the MIC and modulation assays. The antibiotics, amikacin, gentamicin and neomycin, and the antifungals, amphotericin B, benzoylmetronidazole and nystatin, were used in concentrations ranging between 2500 and 2.5?μg/mL. The phytochemical analysis of the EOCL was performed through gas chromatography coupled to a mass spectrometer (GC/MS).

Results: Only Staphylococcus aureus was inhibited by a clinically relevant concentration of EOCL (MIC 512?μg/mL). Synergism between the EOCL and amikacin against S. aureus (9.76?μg/mL) and E. coli (39.062?μg/mL); neomycin against E. coli (2.44?μg/mL); and benzoylmetronidazole against C. krusei (256?μg/mL) were observed. The GC/MS analysis identified cedrol, eucalyptol and α-pinene as the main compounds of EOCL.

Conclusion: EOCL inhibited the growth of S. aureus and potentiated the antibiotic and antifungal effects of drugs against all bacterial and Candida strains, respectively.     

Screening of in vitro antimicrobial activity of plants used in traditional Indonesian medicine

Context: In many regions of Indonesia, there are numerous traditional herbal preparations for treatment of infectious diseases. However, their antimicrobial potential has been poorly studied by modern laboratory methods.

Objective: This study investigates in vitro antimicrobial activity of 49 ethanol extracts from 37 plant species used in Indonesian traditional medicine for treatment against Candida albicans, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus.

Materials and methods: The plants were collected from the Biopharmaca collection garden, Bogor, Indonesia. The plant material was dried, finely grounded, extracted using ethanol, concentrated, and the dried residue was dissolved in 100% DMSO. Antimicrobial activity was determined in terms of a minimum inhibitory concentration (MIC) using a broth microdilution method in 96-well microplates.

Results: The extract of Orthosiphon aristatus (Blume) Miq. (Lamiaceae) leaf produced the strongest antimicrobial effect, inhibiting the growth of C. albicans (MIC 128?μg/mL), S. aureus (MIC 256?μg/mL), E. faecalis (MIC 256?μg/mL) and P. aeruginosa (MIC 256?μg/mL). The leaf extract of Woodfordia floribunda Salisb. (Lythraceae) also exhibited significant effect against C. albicans (MIC 128?μg/mL), S. aureus (MIC 256?μg/mL) and E. faecalis (MIC 256?μg/mL). Rotheca serrata (L.) Steane &; Mabb. (Lamiaceae) leaf extract inhibited the growth of S. aureus (MIC 256 µg/mL) and C. albicans (MIC 256 µg/mL).

Discussion and conclusions: The leaf extract of O. aristatus and W. floribunda exhibited a significant anti-candidal effect. Therefore, both of these plants can serve as prospective source materials for the development of new anti-candidal agents.     

Anti-cryptococcal activity of ethanol crude extract and hexane fraction from Ocimum basilicum var. Maria bonita: mechanisms of action and synergism with amphotericin B and Ocimum basilicum essential oil

Context: Ocimum basilicum L. (Lamiaceae) has been used in folk medicine to treat headaches, kidney disorders, and intestinal worms.

Objective: This study evaluates the anti-cryptococcal activity of ethanol crude extract and hexane fraction obtained from O. basilicum var. Maria Bonita leaves.

Materials and methods: The MIC values for Cryptococcus sp. were obtained according to Clinical and Laboratory Standards Institute in a range of 0.3–2500?μg/mL. The checkerboard assay evaluated the association of the substances tested (in a range of 0.099–2500?μg/mL) with amphotericin B and O. basilicum essential oil for 48?h. The ethanol extract, hexane fraction and associations in a range of 0.3–2500?μg/mL were tested for pigmentation inhibition after 7?days of treatment. The inhibition of ergosterol synthesis and reduction of capsule size were evaluated after the treatment with ethanol extract (312?μg/mL), hexane fraction (78?μg/mL) and the combinations of essential oil?+?ethanol extract (78?μg/mL?+?19.5?μg/mL, respectively) and essential oil?+?hexane fraction (39.36?μg/mL?+?10?μg/mL, respectively) for 24 and 48?h, respectively.

Results: The hexane fraction presented better results than the ethanol extract, with a low MIC (156?μg/mL against C. neoformans T₄₄₄ and 312?μg/mL against C. neoformans H99 serotype A and C. gattii WM779 serotype C). The combination of the ethanol extract and hexane fraction with amphotericin B and essential oil enhanced their antifungal activity, reducing the concentration of each substance needed to kill 100% of the inoculum. The substances tested were able to reduce the pigmentation, capsule size and ergosterol synthesis, which suggest they have important mechanisms of action.

Conclusions: These results provide further support for the use of ethanol extracts of O. basilicum as a potential source of antifungal agents.     

Chemical composition and pharmacological properties of the essential oils obtained seasonally from Lippia thymoides

Context: Lippia thymoides Mart. & Schauer (Verbenaceae) is used in folk medicine to treat wounds, fever, bronchitis, rheumatism, headaches, and weakness.

Objective: This study determinates the chemical composition of essential oils from L. thymoides, obtained at during each of the four seasons and correlates with pharmacological properties.

Materials and methods: Essential oils were obtained by hydrodistillation and analyzed by gas chromatography coupled to mass spectroscopy (GC-MS). Antioxidant activity was determined by DPPH free radical scavenging and β-carotene bleaching methods. The antimicrobial assays were performed by minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) methods. Isolated rat aorta and uterus, and guinea-pig trachea were utilized to evaluate relaxant potential in pre-contracted smooth muscle.

Results and discussion: Essential oils from leaves of L. thymoides had the sesquiterpene β-caryophyllene (17.22–26.27%) as the major constituent followed by borneol (4.45–7.36%), camphor (3.22–8.61%), camphene (2.64–5.66%), and germacrene D (4.72–6.18%). In vitro assays showed that these essential oils do not have antioxidant activity, have antimicrobial selectivity to Gram-positive bacteria Staphylococcus aureus (MIC?=?0.004?mg/mL and MMC?=?0.26–10.19?mg/mL) and Micrococcus luteus (MIC?=?0.03?mg/mL and MMC?=?8.43?mg/mL), relax isolated rat aorta (EC₅₀?=?305–544?μg/mL, with endothelium; and EC₅₀?=?150–283?μg/mL, without endothelium), and uterus (EC₅₀?=?74–257?μg/mL), and minor potency, isolated guinea-pig trachea.

Conclusions: Lippia thymoides is a source of natural products of pharmaceutical interest, being necessary additional studies to determine the substances involved in the biological activities.     

Antibacterial and modulatory effect of Stryphnodendron rotundifolium

Context: Stryphnodendron rotundifolium Mart. (Leguminosae), a tree in Northeast Brazil (Chapada do Araripe), is used in popular medicine to treat different processes such as inflammation and infectious diseases, mainly caused by bacterial pathogens.

Objective: This study determined the modulatory and antimicrobial activity of the hydroethanol extract of dried stem bark, the most used form of this natural product, as a remedy by the traditional communities, against standard and clinical isolates of Escherichia coli and Staphylococcus aureus.

Material and methods: The antibacterial and modulatory activities of the hydroalcoholic extract from the leaves were obtained by maceration/hydrodistillation method and assayed by microdilution.

Results: In the microbiological assays, growth inhibition was demonstrated by this extract against the bacterial strains tested, with minimal inhibitory concentration (MIC) values of 512 µg/mL. However, when a subinhibitory concentration (MIC/8?=?64 µg/mL) was combined with conventional antimicrobial drugs (gentamicin, kanamycin, amikacin and neomycin), the extract showed a potentiating effect, reducing the MIC for all drugs assayed in a range between 312.5 and 2.4 µg/mL.

Conclusions: We indicate that the extract of S. rotundifolium showed potential synergistic antibiotic activity. With the results obtained, these extracts proved to be a promising source of antibacterial and modulatory agents.     

Ocimum basilicum: Antibacterial activity and association study with antibiotics against bacteria of clinical importance

Context: Ocimum basilicum L. (Lamiaceae), popularly known as basil, is part of a group of medicinal plants widely used in cooking and known for its beneficial health properties, possessing significant antioxidant effects, antinociceptive, and others.

Objective: The objective of this study is to determine the pharmacological effects produced on the bacterial strains Staphylococcus aureus and Pseudomonas aeruginosa when standard antibiotics and O. basilicum essential oil are combined.

Materials and methods: The extraction of O. basilicum (leaves) components was done by steam distillation. The Minimum inhibitory concentration (MIC) was calculated using microdilution technique, where the oil concentrations varied from 2 to 1024?μg/mL. The combinations of O. basilicum oil with ciprofloxacin or imipenem were analyzed by the checkerboard method where fractional inhibitory concentration (FIC) indices were calculated.

Results: Ocimum basilicum essential oil, imipenem, and ciprofloxacin showed respective MIC antibacterial activities of 1024, 4, and 2?μg/mL, against S. aureus. In S. aureus, the oil with imipenem association showed synergistic effect (FIC?=?0.0625), while the oil with ciprofloxacin showed antagonism (FIC value?=?4.25). In P. aeruginosa, the imipenem/oil association showed additive effect for ATCC strains, and synergism for the clinical strain (FIC values?=?0.75 and 0.0625). The association of O. basilicum essential oil with ciprofloxacin showed synergism for clinical strains (FIC value?=?0.09).

Conclusion: Ocimum basilicum essential oil associated with existing standard antibiotics may increase their antibacterial activity, resulting in a synergistic activity against bacterial strains of clinical importance. The antibacterial activity of O. basilicum essential oil may be associated with linalool.     

“Aroeira” (Myracrodruon urundeuva) methanol extract: the relationship between chemical compounds and cellular effects

Context: “Aroeira” [Myracrodruon urundeuva Allemão (Anacardiaceae)] is a tree whose leaves have been studied for therapeutic purposes in medicine and dentistry.

Objective: The study chemically identifies the leaf extract of aroeira and determines its effect on human gingival fibroblasts.

Materials and methods: An 80% methanol leave extract was obtained by maceration and chemically identified through flow-injection analysis–electrospray ionization–ion trap–tandem mass spectrometry (FIA–ESI–IT–MSⁿ). Cytotoxicity of the aroeira’s methanol extract was evaluated in lineage of fibroblasts. Adherent cells were treated with different concentrations of aroeira’s methanol extract in the medium: 0.1, 1, 10, 100 and 1000?μg/mL. Control cells were cultivated in the medium only. Analyses were done at 24, 48, 72 and 96?h of culture by neutral red assay; and at 24, 48 and 96?h by crystal violet assay.

Results: FIA–ESI–IT–MS analysis determined the presence of compounds, for the first time in the species: quercetin-O-glucuronide and quercetin-O-deoxyhexose-O-glucose in the extract. On one hand, neutral red and crystal violet assay showed a reduction (to 50% up until 100%) of cellular viability of groups of 100 and 1000?μg/mL compared with control at 96?h (p?<?0.05). On the other hand, lower concentrations (0.1; 1 and 10?μg/mL) of the extract were similar to that of the control at 96?h (p?<?0.05), in general.

Conclusions: In view of the results, we can conclude that the extract of aroeira presents tannins and flavonoids. Furthermore, the extract is capable of modulating the viability of human gingival fibroblasts according to its concentration.     

Potential anti-inflammatory,antioxidant and antimicrobial activities of Sambucus australis

Context: Sambucus australis Cham. &; Schltdl. (Adoxaceae) is used in Brazilian folk medicine to treat inflammatory disorders.

Objective: To evaluate the in vitro anti-inflammatory, antioxidant and antimicrobial properties of S. australis.

Materials and methods: The anti-in?ammatory activity of ethanol extracts of the leaf and bark of S. australis (1–100?μg/mL) were studied in lipopolysaccharide/interferon γ stimulated murine macrophages RAW 264.7 cells (24?h incubation) by investigating the release of nitric oxide (NO) and tumour necrosis factor-alpha (TNF-α) and in the TNF-α-induced nuclear factor kappa (NF-κB) assay. Minimum inhibitory concentration (MIC) was determined by the microdilution test (24?h incubation). Antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and the NO scavenging assays. Chemical composition was assessed by LC-MS/MS.

Results: Antioxidant activities in the DPPH (IC₅₀ 43.5 and 66.2?μg/mL), FRAP (IC₅₀ 312.6 and 568.3?μg/mL) and NO radical scavenging assays (IC₅₀ 285.0 and 972.6?μg/mL) were observed in the leaf and bark ethanol extracts, respectively. Solely the leaf extract showed significant inhibition of NO and TNF-α production in RAW264.7 cells at concentrations of 2 and 100?μg/mL, respectively, and suppression of TNF-α inhibition of NF-κB by 12.8 and 20.4% at concentrations of 50 and 100?μg/mL, respectively. The extract also exhibited antibacterial activity against Salmonella typhimurium (MIC 250?μg/mL) and Klebsiella pneumoniae (MIC 250?μg/mL). LC-MS/MS revealed the presence of chlorogenic acid and rutin as major compounds.

Discussion and conclusion: The results indicate that the ethanol leaf extract of S. australis exhibit prominent anti-in?ammatory effects.     

Preliminary studies on the antibacterial activity of crude extracts and alkaloids from species of Aspidosperma

Screening tests of hydroethanolic crude extracts of six species of Aspidosperma (Apocynaceae) against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Pseudomonas aeruginosa were performed. Aspidosperma ramiflorum Muell. Arg. showed good activity against Bacillus subtilis with MIC and MBC of 15.7 and 125?μg/mL, moderate activity against Staphylococcus aureus with MIC and MBC of 250 and 500?μg/mL, and weak activity against Escherichia coli with MIC and MBC of 1000?μg/mL. Aspidosperma pyricolum Muell. Arg. (MIC/MBC 125/250?μg/mL) and Aspidosperma olivaceum Muell. Arg. (MIC/MBC 250/?>?1000?μg/mL) displayed moderate antibacterial activity against Bacillus subtilis. Separation of the crude extract of Aspidosperma ramiflorum was performed according to the usual acid–base process, which produces alkaloid mixtures and closely related metabolites. The basic fraction was active against Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, with MICs of 31.2, 62.5, and 250?μg/mL, respectively. The basic fractions were more active than the acid fractions, probably because they contained some active alkaloids and/or closely related metabolites absent from the other fractions, or they contained a higher concentration of these active compounds.     

Bioactive compounds from Hypericum humifusum and Hypericum perfoliatum: inhibition potential of polyphenols with acetylcholinesterase and key enzymes linked to type-2 diabetes

Context: Natural products are reported to have a wide spectrum of pharmacological properties such as antimicrobial, anti-inflammatory and anti-cholinesterase. The genus Hypericum (Hypericaceae) is a source of a variety of molecules with different biological activities, notably hypericin and various phenolics.

Objectives: The goals of the present work were the determination of total phenolic and flavonoid content, hypericin and hyperforin concentration as well as the evaluation of biological of Hypericum humifusum L. (Hhu) and Hypericum perfoliatum L. (Hper).

Materials and methods: The various extracts of aerial parts were powdered, and then extracted with methanol. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) methods against four Gram-positive bacteria, four Gram-negative bacteria and yeast.

Results: The results revealed that H. humifusum, bear the highest total phenolic and flavonoid content (48–113?mg GAE/g and 8–41?mg RE/g, respectively) as well as hypericin (60–90?mg/g) and hyperforin (8–30?mg/g) concentration. Both species showed significant antioxidant activity as revealed by DPPH, FRAP, ABTS, and metal chelating assays. H. humifusum exhibited a strong acetylcholinesterase (3.86–4.57?mg GALAEs/g), α-glucosidase (0.73–2.55?mmol ACEs/g) and α-amylase (3–8?mmol ACEs/g) inhibitory activity. The extract of H. humifusum exhibited strong antibacterial activity mainly against Staphylococcus epidermidis, Staphylococus aureus, and Enterococcus faecium (MIC values ranging from 200 to 250?μg/mL). The highest antifungal activity was showed for H. perfoliatum extract (MIC value = 250?μg/mL).

Conclusion: The data suggest that H. humifusum could be used as valuable new natural agents with functional properties for pharmacology industries.     

Antimicrobial activity of pomegranate fruit constituents against drug-resistant Mycobacterium tuberculosis and β-lactamase producing Klebsiella pneumoniae

Abstract

Context: The global surge in multi-drug resistant bacteria and the imminence of tuberculosis pandemic necessitate alternative therapeutic approaches to augment the existing medications. Pomegranate, the fruit of Punica granatum Linn. (Punicaceae), widely recognized for potency against a broad spectrum of bacterial pathogens, deserves further investigation in this respect.

Objective: This study determines the therapeutic potential of pomegranate juice, extracts of non-edible peel prepared with methanol/water, and its four polyphenolic constituents, namely caffeic acid, ellagic acid, epigallocatechin-3-gallate (EGCG) and quercetin, against drug-resistant clinical isolates.

Materials and methods: Phenotypic characterisation of Mycobacterium tuberculosis, extended-spectrum β-lactamase (ESBL) and KPC-type carbapenemase producing Klebsiella pneumoniae was performed by biochemical and molecular methods. Resistance profiles of M. tuberculosis and K. pneumoniae were determined using LJ proportion and Kirby–Bauer methods, respectively. Pomegranate fruit extracts, and the compounds, were evaluated at a dose range of 1024–0.5?µg/mL, and 512–0.25?µg/mL, respectively, to determine minimum inhibitory (MIC) and bactericidal concentrations (MBC) against the drug-resistant isolates by the broth micro-dilution method.

Results: The peel extracts exhibited greater antimycobacterial activity (MIC 64–1024?μg/mL) than the potable juice (MIC 256?-?>?1024?μg/mL). EGCG and quercetin exhibited higher antitubercular (MIC 32–256?μg/mL) and antibacterial (MIC 64–56?μg/mL) potencies than caffeic acid and ellagic acid (MIC 64–512?μg/mL).

Discussion and conclusion: The pomegranate fruit peel and pure constituents were active against a broad panel of M. tuberculosis and β-lactamase producing K. pneumoniae isolates. EGCG and quercetin need further investigation for prospective application against respiratory infections.     

Bioactive potential of endophytic Myrothecium sp. isolate M1-CA-102, associated with Calophyllum apetalum

Context: Endophytes colonizing medicinal plants are diverse, constituting a rich bioresource for novel natural products.

Objective: Myrothecium sp. isolate M1-CA-102 was the most promising among the 16 Myrothecium isolates screened. The bioactive potential of the crude extract from the Calophyllum apetalum Willd. endophytic Myrothecium sp. (Alb. &; Schwein.) Ditmar (Incertae sedis) isolate M1-CA-102 and its thin layer chromatography (TLC) fractions were screened based on antioxidant, anti-inflammatory, antimicrobial activities, and cytotoxicity.

Materials and methods: The antioxidant activity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging capacities. Further, 15-lipoxygenase (15-LOX) and human cyclooxygenase-2 (COX-2) inhibition were assessed at different concentrations (25, 50, and 100?μg/mL for the crude extract, 5, 25, and 50?μg/mL for the TLC fractions). DNA-nicking assay as an indicator of the capacity of extracts to scavenge hydroxyl radical was recorded at a concentration of 50?μg/mL. Cell cytotoxicity was recorded by colorimetric 3-(4,5-dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Antibacterial (Bacillus subtilis) and anti-Candida (Candida albicans) assays were performed by the microdilution method.

Results: The DPPH and ABTS IC₅₀ values of M1-CA-102 extract were 10 and 6?μg/mL compared with 6.1 and 7.03?μg/mL for the positive control quercetin. The cytotoxicity IC₅₀ value of M1-CA-102 extract was 37?μg/mL, while the M-I TLC fraction was 21?μg/mL. The M1-CA-102 extract gave an IC₅₀ value of 58 and 8?μg/mL for 15-LOX and COX-2, respectively. The MIC values for antimicrobial activity for M1-CA-102 extract ranged from 35 to 54?μg/mL, while for the TLC fractions, it ranged from 91 to 515?μg/mL.

Conclusion: The results indicate that Myrothecium M1-CA-102 isolated from C. apetalum is a potential source of natural metabolites of pharmaceutical importance.     

Chemical composition along with anti-leishmanial and cytotoxic activity of Zataria multiflora

Context: Natural products and their compounds are some of the most interesting sources of new drugs. Reviews have reported various pharmacological properties such as antimicrobial effects of Zataria multiflora Boiss (Lamiaceae).

Objective: The present study investigates the chemical composition of Z. multiflora essential oil and evaluates its cytotoxic effects and anti-leishmanial activities against Leishmania tropica in an in vitro model.

Materials and methods: The components of Z. multiflora oil were identified by gas chromatography/mass spectroscopy (GC/MS) analysis. Anti-leishmanial effects of the essential oil (0–100?μL/mL) and methanol extract of Z. multiflora (0–100 μg/mL) on promastigote forms as well as their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 h. The leishmanicidal activity against amastigote forms of L. tropica was evaluated at the concentrations of 0–50 μg/mL in a macrophage model for 48 h.

Results: The chemical analyses demonstrated that the main components of essential oil were thymol (41.81%), carvacrol (28.85%), and p-cymene (8.36%). Regarding leishmanicidal activity, the IC₅₀ values for the essential oil and methanol extract were 3.2?μL/mL and 9.8 μg/mL against promastigote forms and 8.3?μL/mL and 34.6 μg/mL against amastigote forms, respectively. Essential oil (CC₅₀ 89.3?μL/mL) indicated a higher cytotoxic effect than the methanol extract (CC₅₀ 591.6 μg/mL) of Z. multiflora.

Conclusion: The present study revealed the chemical composition of Z. multiflora that might be a natural source of new anti-leishmanial agents in terms of use against cutaneous leishmaniasis.     

The anti-dermatophyte activity of Commiphora molmol

Context: Commiphora molmol Engl (Burseraceae) or myrrh has been traditionally used for the treatment of skin fungal infections.

Objective: This study evaluates the antifungal activity of myrrh ethanol extract and essential oil against skin dermatophytes.

Materials and methods: The antifungal evaluations were performed by the food poisoning technique (250?ppm) and micro-broth dilution assay (800–6.25?µg/mL) against Trichophyton rubrum, T. mentagrophytes, Microsporum canis, M. gypseum, and T. verrucosum. The chemical composition of myrrh oil and ethanol extract was analyzed by GC and GC-MS.

Results: Furanoeudesma 1,3-diene and menthofuran were the main components of myrrh oil, while 2-tert-butyl-1,4-naphthoquinone, benzenemethanol,3-methoxy-α-phenyl, and curzerene were the main components of myrrh ethanol extract. The inhibitory effect of myrrh oil and ethanol extract against dermatophytes were 43.1–61.6% and 12.5–27.5%, respectively. The MIC and MFC values of myrrh oil were 25–100 and 25–200?µg/mL while these amounts for ethanol extract were 25–400 and 25–400?µg/mL, respectively. Therefore, myrrh oil had higher antifungal activity than that of the ethanol extract. Both extracts showed good anti-elastase activity.

Conclusion: The results of our investigation confirmed the traditional uses of C. molmol as a poultice for the treatment of cutaneous fungal infections.     

Leishmanicidal and cytotoxic activities of Nigella sativa and its active principle,thymoquinone

Abstract

Context: Leishmaniasis is a complex disease with a broad spectrum of clinical presentations.

Objective: We evaluated the anti-leishmanial effects of Nigella sativa L. (Ranunculaceae) against Leishmania tropica and Leishmania infantum with an in vitro model.

Materials and methods: Antileishmanial effects of essential oil and methanolic extract of N. sativa (0–200?µg/mL) and thymoquinone (0–25?µg/mL) on promastigotes of both species and their cytotoxicity activities against murine macrophages were evaluated using the MTT assay at 24, 48, and 72?h. Moreover, their leishmanicidal effects against amastigotes were investigated in a macrophage model, for 48 and 72?h.

Results: The findings showed that essential oil (L. tropica IC₅₀ 9.3?μg/mL and L. infantum IC₅₀ 11.7?μg/mL) and methanolic extract (L. tropica IC₅₀ 14.8?μg/mL and L. infantum IC₅₀ 15.7?μg/mL) of N. sativa, particularly thymoquinone (L. tropica IC₅₀ 1.16?μg/mL and L. infantum IC₅₀ 1.47?μg/mL), had potent antileishmanial activity on promastigotes of both species after 72?h. In addition, essential oil (L. tropica IC₅₀ 21.4?μg/mL and L. infantum IC₅₀ 26.3?μg/mL), methanolic extract (L. tropica IC₅₀ 30.8?μg/mL and L. infantum IC₅₀ 34.6?μg/mL), and thymoquinone (L. tropica IC₅₀ 2.1?μg/mL and L. infantum IC₅₀ 2.6?μg/mL) mediated a significant decrease in the growth rate of amastigote forms of both species. Thymoquinone (CC₅₀ 38.8?μg/mL) exhibited higher cytotoxic effects against murine macrophages than the other extracts.

Conclusion: N. sativa, especially its active principle, thymoquinone, showed a potent leishmanicidal activity against L. tropica and L.infantum with an in vitro model.     

Anti-acetylcholinesterase activity and antioxidant properties of extracts and fractions of Carpolobia lutea

Context: There is an unmet need to discover new treatments for Alzheimer’s disease. This study determined the anti-acetylcholinesterase (AChE) activity, DPPH free radical scavenging and antioxidant properties of Carpolobia lutea G. Don (Polygalaceae).

Objective: The objective of this study is to quantify C. lutea anti-AChE, DPPH free radical scavenging, and antioxidant activities and cell cytotoxicity.

Materials and methods: Plant stem, leaves and roots were subjected to sequential solvent extractions, and screened for anti-AChE activity across a concentration range of 0.02–200?μg/mL. Plant DPPH radical scavenging activity, reducing power, and total phenolic and flavonoid contents were determined, and cytotoxicity evaluated using human hepatocytes.

Results: Carpolobia lutea exhibited concentration-dependent anti-AChE activity. The most potent inhibitory activity for the stem was the crude ethanol extract and hexane stem fraction oil (IC₅₀?=?140?μg/mL); for the leaves, the chloroform leaf fraction (IC₅₀?=?60?μg/mL); and for roots, the methanol, ethyl acetate and aqueous root fractions (IC₅₀?=?0.3–3?μg/mL). Dose-dependent free radical scavenging activity and reducing power were observed with increasing stem, leaf or root concentration. Total phenolic contents were the highest in the stem: ～632?mg gallic acid equivalents/g for a hexane stem fraction oil. Total flavonoid content was the highest in the leaves: ～297?mg quercetin equivalents/g for a chloroform leaf fraction. At 1?μg/mL, only the crude ethanol extract oil was significantly cytotoxic to hepatocytes.

Discussion and conclusions: Carpolobia lutea possesses anti-AChE activity and beneficial antioxidant capacity indicative of its potential development as a treatment of Alzheimer’s and other diseases characterized by a cholinergic deficit.     

In vitro antimicrobial and anti-proliferative activities of plant extracts from Spathodea campanulata,Ficus bubu,and Carica papaya

Context African medicinal plants represent a prominent source of new active substances. In this context, three plants were selected for biological investigations based on their traditional uses.

Objective The antimicrobial and anti-proliferative features of three plants used for medicinal purpose were evaluated.

Materials and methods The antimicrobial activities of methanol extracts of Ficus bubu Warb. (Moraceae) stem bark and leaves, of Spathodea campanulata P. Beauv. (Bignoniaceae) flowers, as well as those of Carica papaya Linn. (Caricaceae) latex, were determined using the microbroth dilution method against a set of bacteria and fungi pathogens including: Enterococcus faecalis, Staphylococcus aureus, S. saprophyticus, S. epidermididis, Escherichia coli, Klebsiella pneumonia, Salmonella typhimurium, Candida albicans, and Trichophyton rubrum. The tested concentrations of extracts ranged from 2500.0 to 2.4?μg/mL and MIC values were evaluated after 24?h incubation at 37?°C. Subsequently, MTT assay was used to estimate anti-proliferative activity of these methanol extracts and of F. bubu latex on three human cancer cell lines (U373 glioblastoma, A549 NSCLC, and SKMEL-28 melanoma).

Results The methanol extract of F. bubu stem bark exhibited the highest antimicrobial activity against C. albicans with a MIC value of 9.8?μg/mL, while the F. bubu latex and the methanol extract of F. bubu leaves induced significant anti-proliferative activity against lung (IC₅₀ values of 10 and 14?μg/mL, respectively) and glioma (IC₅₀ values of 13 and 16?μg/mL, respectively) cancer cells.

Conclusion These results indicate that effective drugs could be derived from the three studied plants.     

Using rifapentine – hen egg lipoprotein conjugate as macrophage-targeted drug delivery carrier against intracellular Staphylococcus aureus

Abstract

Context: Hen egg low-density lipoprotein (heLDL), which is present in large quantities in egg yolk, share a high identity with human apolipoprotein B-100 precursor.

Objective: This study investigated the use of heLDL as a macrophage-targeted drug delivery carrier against intracellular Staphylococcus aureus.

Methods: Rifapentine (RPT) was incorporated into heLDL (RPT–heLDL). Staphylococcus aureus ATCC 29740 and human U937 macrophage were used as intracellular infection models.

Results and conclusion: The loading efficiency of RPT into the heLDL was 66.10?±?2.28?μg RPT/mg heLDL. Fluorescence microscopy and oil red O staining results indicated RPT–heLDL can be taken up by U937 macrophages. The cell viability (MTT assay) was increased when the concentration of heLDL was <150?μg/mL. Unloaded heLDL (100?μg/mL) can inhibit the growth of intracellular S. aureus compared with the untreated control group after 18?h incubation. RPT–heLDL (6.6?μg/mL RPT, 100?μg/mL heLDL) eliminated 94% of intracellular S. aureus, whereas the corresponding dose of free RPT (6.6?μg/mL) induced an 87% reduction. The in vitro results of the current study indicated that heLDL might be used as a suitable drug carrier for targeting human macrophages.     

Unlocking the in vitro anti-inflammatory and antidiabetic potential of Polygonum maritimum

Context: Several Polygonum species (Polygonaceae) are used in traditional medicine in Asia, Europe and Africa to treat inflammation and diabetes.

Objective: Evaluate the in vitro antioxidant, anti-inflammatory and antidiabetic potential of methanol and dichloromethane extracts of leaves and roots of the halophyte Polygonum maritimum L.

Material and methods: Antioxidant activity was determined (up to 1?mg/mL) as radical-scavenging activity (RSA) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), copper (CCA) and iron (ICA) chelating activities and iron reducing power (FRAP). NO production was measured in lipopolysaccharide (LPS)-stimulated macrophages for 24?h at concentrations up to 100?μg/mL and antidiabetic potential was assessed by α-amylase and α-glucosidase inhibition (up to 10?mg/mL) assays. The phytochemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS).

Results: The methanol leaf extract had the highest activity against DPPH? (IC₅₀ =?26?μg/mL) and ABTS⁺? (IC₅₀ =?140?μg/mL), FRAP (IC₅₀ =?48?μg/mL) and CCA (IC₅₀ =?770?μg/mL). Only the dichloromethane leaf extract (LDCM) showed anti-inflammatory activity (IC₅₀ =?48?μg/mL). The methanol root (IC₅₀ =?19?μg/mL) and leaf (IC₅₀ =?29?μg/mL) extracts strongly inhibited baker’s yeast α-glucosidase, but LDCM had higher rat’s α-glucosidase inhibition (IC₅₀ =?2527?μg/mL) than acarbose (IC₅₀ =?4638?μg/mL). GC-MS analysis identified β-sitosterol, stigmasterol, 1-octacosanol and linolenic acid as possible molecules responsible for the observed bioactivities.

Conclusions: Our findings suggest P. maritimum as a source of high-value health promoting commodities for alleviating symptoms associated with oxidative and inflammatory diseases, including diabetes.     

Chemical composition,antioxidant and antibacterial activities of essential oils from Ferulago angulata

Context: Ferulago angulata Boiss. (Apiaceae), a perennial aromatic herb, grows wild in Iran. The aerial parts of F. angulata are used as a flavouring in foods, especially dairy foods by indigenous people in western and southwestern Iran.

Objective: This study investigates variation in chemical compositions, antioxidant and antibacterial activities of the essential oils from F. angulata collected from natural habitats in the alpine regions of southwestern Iran.

Materials and Methods: The antimicrobial activity, minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) of the essential oils were evaluated against four bacteria (Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and Salmonella typhimurium). Antioxidant activity of the oils was determined by DPPH assay.

Results: The essential oils were analyzed by GC-FID and GC/MS, which 49 volatile components were identified. There were significant differences between the various populations for oil yield and some main compounds. The major constituents of the essential oils from F. angulata were α-pinene, and cis-β-ocimene. The MICs of the essential oils were within concentration ranges from 62 to 250?μg/mL and the respective MBCs were 125 to?>?500?μg/mL. Generally, the oils from F. angulata indicated weak to moderate inhibitory activities against bacteria, especially against Listeria monocytogenes. The highest antioxidant activity was obtained from the oil of the Kallar population (IC₅₀ value_?=_?488?μg/mL) and BHT as positive control (IC_50? value =_?321?μg/mL).

Discussion and conclusion: The essential oil of F. angulata could be serving as a potential source of α-pinene and cis-β-ocimene for use in the food, cosmetic and pharmaceutical industries.