Carvacrol as a Stimulant of the Expression of Key Genes of the Ginsenoside Biosynthesis Pathway and Its Effect on the Production of Ginseng Saponins in Panax quinquefolium Hairy Root Cultures.

The accumulation of ginsenosides (triterpenic saponins) was determined in Panax quinquefolium hairy root cultures subjected to an elicitation process using carvacrol at 5, 10, 25, 50, 100, 250, and 500 µM concentrations during 24 and 72 h exposure. This study was the first one in which carvacrol was applied as an elicitor. The content of eight ginsenosides, Rb1, Rb2, Rb3, Rc, Rd, Rg1, Rg2, and Re, was determined using HPLC analysis. Moreover, the quantitative RT-PCR method was applied to assess the relative expression level of farnesyl diphosphate synthase, squalene synthase, and dammarenediol synthase genes in the studied cultures. The addition of carvacrol (100 µM) was an effective approach to increase the production of ginsenosides. The highest content and productivity of all detected saponins were, respectively, 20.01 mg∙g-1 d.w. and 5.74 mg∙L-1∙day-1 after 72 h elicitation. The production profile of individual metabolites in P. quinquefolium cultures changed under the influence of carvacrol. The biosynthesis of most examined protopanaxadiol derivatives was reduced under carvacrol treatment. In contrast, the levels of ginsenosides belonging to the Rg group increased. The strongest effect of carvacrol was noticed for Re metabolites, achieving a 7.72-fold increase in comparison to the control. Saponin Rg2, not detected in untreated samples, was accumulated after carvacrol stimulation, reaching its maximum concentration after 72 h exposure to 10 µM elicitor.

Evaluation of antioxidant activity of NG-R1 saponin against bacterial cells in induced oxidative stress - a preliminary study.

INTRODUCTION AND OBJECTIVE: Notoginsenoside R1 (NG-R1) is isolated from Panax notoginseng, a medicinal herb well-known for its long use in traditional Chinese medicine. NG-R1 is relatively under-studied in research on bacteria. The aim of the study was to investigate antioxidant properties of NG-R1 saponin of selected bacterial strains of intestinal microbiota that may be involved in the pathogenesis of thromboembolic diseases. Enterococcus faecalis and Escherichia coli were used in the study. MATERIAL AND METHODS: The study determined the concentration of hydroperoxides, the level of lipid peroxidation, as well as carbonyl groups and free thiol groups. The research carried out in this way will allow determination of the influence of the above factors on bacteria living in intestinal microbiota. RESULTS: An evaluation of selected parameters of oxidative stress allowed to check whether the tested compound could reduce the pro-thrombotic activity of bacteria that were stimulated with H2O2. It was found that NG-R1 reduced hydroperoxide levels in both types of bacteria. In turn, lipid peroxidation initiated by H2O2 was suppressed by NG-R1. Hydrogen peroxide led to a strong increase in the level of carbonyl groups in Enterococcus faecalis and, to a lesser extent, in Escherichia coli. The addition of NG-R1 to the medium significantly reduced the level of carbonyls. Additionally, NG-R1 also induced a significant increase in the level of free thiol groups. CONCLUSIONS: Obtained results indicate that NG-R1 may have a protective effect on the intestinal microbiom through mechanisms involving changes in the redox state.

The Effect of Fermentation with Kefir Grains on the Physicochemical and Antioxidant Properties of Beverages from Blue Lupin (Lupinus angustifolius L.) Seeds.

Plant derived fermented beverages have recently gained consumers' interest, particularly due to their intrinsic functional properties and presence of beneficial microorganisms. Three variants containing 5%, 10%, and 15% (w/w) of sweet blue lupin (Lupinus angustifolius L. cv. "Boregine") seeds were inoculated with kefir grains and incubated at 25 °C for 24 h. After processing, beverages were stored in refrigerated conditions (6 °C) for 21 days. Changes in microbial population, pH, bioactive compounds (polyphenolics, flavonoids, ascorbic acid), reducing sugars, and free amino acids were estimated. Additionally, viscosity, firmness, color, and free radicals scavenging properties were determined. Results showed that lactic acid bacteria as well as yeast were capable of growing well in the lupin matrix without any supplementation. During the process of refrigeration, the viability of the microorganisms was over the recommended minimum level for kefir products. Hydrolysis of polysaccharides as well as increase of free amino acids was observed. As a result of fermentation, the beverages showed excellent DPPH, ABTS+·, ·OH, and O2- radicals scavenging activities with a potential when considering diseases associated with oxidative stress. This beverages could be used as a new, non-dairy vehicle for beneficial microflora consumption, especially by vegans and lactose-intolerant consumers.

Development, Characterization, and Bioactivity of Non-Dairy Kefir-Like Fermented Beverage Based on Flaxseed Oil Cake.

Flaxseed oil cake (FOC) was evaluated as a potential substrate for the production of a novel kefir-like fermented beverage. Three variants containing 5%, 10%, and 15% (w/w) of FOC were inoculated with kefir grains and incubated at 25 °C for 24 h. After processing, beverages were stored in refrigerated conditions (6 °C) for 21 days. Changes in microbial population, pH, acidity, levels of proteins, polyphenolics, flavonoids, ascorbic acid, and reducing sugars were estimated. Additionally, viscosity, firmness, color, and antioxidant properties were determined. Results showed that lactic acid bacteria as well as yeast were capable of growing well in the FOC without any supplementation. During refrigerated storage, the viability of the microorganisms were over the recommended minimum level for kefir products. As a result of fermentation, the beverages showed excellent antioxidant activity. Because of the functional characteristics conferred to the FOC beverages, the use of kefir grains showed adequate potential for the industrial application. Therefore, this beverages could be used as a new, non-dairy vehicle for beneficial microflora consumption, especially by vegans and lactose-intolerant consumers.

Abscisic Acid Regulates the 3-Hydroxy-3-methylglutaryl CoA Reductase Gene Promoter and Ginsenoside Production in Panax quinquefolium Hairy Root Cultures.

Panax quinquefolium hairy root cultures synthesize triterpenoid saponins named ginsenosides, that have multidirectional pharmacological activity. The first rate-limiting enzyme in the process of their biosynthesis is 3-hydroxy-3-methylglutaryl CoA reductase (HMGR). In this study, a 741 bp fragment of the P. quinquefolium HMGR gene (PqHMGR), consisting of a proximal promoter, 5'UTR (5' untranslated region) and 5'CDS (coding DNA sequence) was isolated. In silico analysis of an isolated fragment indicated a lack of tandem repeats, miRNA binding sites, and CpG/CpNpG elements. However, the proximal promoter contained potential cis-elements involved in the response to light, salicylic, and abscisic acid (ABA) that was represented by the motif ABRE (TACGTG). The functional significance of ABA on P. quinquefolium HMGR gene expression was evaluated, carrying out quantitative RT-PCR experiments at different ABA concentrations (0.1, 0.25, 0.5, and 1 mg·L-1). Additionally, the effect of abscisic acid and its time exposure on biomass and ginsenoside level in Panax quinquefolium hairy root was examined. The saponin content was determined using HPLC. The 28 day elicitation period with 1 mg·L-1 ABA was the most efficient for Rg2 and Re (17.38 and 1.83 times increase, respectively) accumulation; however, the protopanaxadiol derivative content decreased in these conditions.

The Antibacterial Activity of Lavender Essential Oil Alone and In Combination with Octenidine Dihydrochloride against MRSA Strains.

In the post-antibiotic era the issue of bacterial resistance refers not only to antibiotics themselves but also to common antiseptics like octenidine dihydrochloride (OCT). This appears as an emerging challenge in terms of preventing staphylococcal infections, which are both potentially severe and easy to transfer horizontally. Essential oils have shown synergisms both with antibiotics and antiseptics. Therefore the aim of this study was to investigate the impact of lavender essential oil (LEO) on OCT efficiency towards methicillin-resistant S. aureus strains (MRSA). The LEO analyzed in this study increased the OCT's susceptibility against MRSA strains. Subsequent FTIR analysis revealed cellular wall modifications in MRSA strain cultured in media supplemented with OCT or LEO/OCT. In conclusion, LEO appears to be a promising candidate for an efficient enhancer of conventional antiseptics.

The Increase of Triterpene Saponin Production Induced by Trans-Anethole in Hairy Root Cultures of Panax quinquefolium.

In vitro cultivation is an effective way to increase pharmaceutical production. To increase ginsenoside production in hairy root cultures of American ginseng, the present study uses trans-anethole as an elicitor. The content of nine triterpene saponins was determined: Rb1, Rb2, Rb3, Rc, Rd, Rg1, Rg2, Re and Rf. Trans-anethole was found to stimulate saponin synthesis regardless of exposure time (24 and 72 h). Twenty-four hour exposure to 1 µmol trans-anethole in the culture medium resulted in the highest increase of total saponin content (twice that of untreated roots), and optimum accumulation of Rb-group saponins, with ginsenoside Rc dominating (8.45 mg g-1 d.w.). In contrast, the highest mean content of protopanaxatriol derivatives was obtained for 10 µmol trans-anethole. The Re metabolite predominated, reaching a concentration of 5.72 mg g-1 d.w.: a 3.9-fold increase over untreated roots. Elicitation with use of trans-anethole can therefore be an effective method of increasing ginsenoside production in shake flasks.

Abies Concolor Seeds and Cones as New Source of Essential Oils-Composition and Biological Activity.

The chemical composition, including the enantiomeric excess of the main terpenes, of essential oils from seeds and cones of Abies concolor was studied by chromatographic (GC) and spectroscopic methods (mass spectrometry, nuclear magnetic resonance), leading to the determination of 98 compounds. Essential oils were mainly composed of monoterpene hydrocarbons. The dominant volatiles of seed essential oil were: limonene (47 g/100 g, almost pure levorotary form) and α-pinene (40 g/100 g), while α-pinene (58 g/100 g), sabinene (11 g/100 g), and ß-pinene (4.5 g/100 g) were the predominant components of the cone oil. The seed and cone essential oils exhibited mild antibacterial activity, and the MIC ranged from 26 to 30 µL/mL against all of the tested bacterial standard strains: Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, and Klebsiella pneumoniae. The cytotoxic studies have demonstrated that tested essential oils were cytotoxic to human skin fibroblasts and human microvascular endothelial cells at concentrations much lower than the MIC. The essential oils from A. concolor seeds and cones had no toxic effect on human skin fibroblasts and human microvascular endothelial cells, when added to the cells at a low concentration (0-0.075 µL/mL) and (0-1.0 µL/mL), respectively, and cultured for 24 h.

The ability of selected plant essential oils to enhance the action of recommended antibiotics against pathogenic wound bacteria.

The aim of this work was to characterize the ability of essential oils to support antibiotics against pathogenic bacteria in wounds. Gram-positive and Gram-negative bacteria obtained from wound infections were identified according to standard microbiological methods. Essential oils were analysed by GC-FID-MS. The susceptibility of bacteria to antibiotics, essential oils and their combination was assessed using the disc-diffusion method. The Minimal Inhibitory Concentration and Minimum Bactericidal Concentration of the essential oils were established by the micro-dilution broth method. Although cinnamon, clove, thyme and lavender essential oils were found to have the greatest antibacterial activity when used alone, the greatest additive and synergistic effects against pathogenic wound bacteria in combination with recommended antibiotics were demonstrated by basil, clary sage and rosemary oils.

Chemical Composition and Antibacterial Activity of the Essential Oil of Phellodendron lavallei.

The essential oils from inflorescences, fruits and leaves of Phellodendron lavallei Dode were analyzed by GC, GC/MS and IH NMR spectroscopy. About 100 compounds were identified. The principal components in the fruit oil were myrcene (47.7-52.0%) and limonene (38.4-40.9%), followed by germacrene D (1.2-3.2%) and (E)-p-caryophyllene (2.4-2.9%). The inflorescence oil contained mainly myrcene (44.8%), limonene (43.2%) and (E)-ß-ocimene (4.1%). The main constituents of the leaf oil were limonene (26.7%) myrcene (22.1%), (E,E)-a-famesene (5.6%) and a-pinene (5.0%). The antibacterial activity was examined towards both Gram-positive and Gram-negative bacteria. The fruit oil showed weak activity against the bacteria tested.

The influence of NDT-Bobath and PNF methods on the field support and total path length measure foot pressure (COP) in patients after stroke.

In stroke patients, the NDT - (Bobath - Neurodevelopmental Treatment) and PNF (Proprioceptive Neuromuscular Facilitation) methods are used to achieve the main objective of rehabilitation, which aims at the restoration of maximum patient independence in the shortest possible period of time (especially the balance of the body). The aim of the study is to evaluate the effect of the NDT-Bobath and PNF methods on the field support and total path length measure foot pressure (COP) in patients after stroke. The study included 72 patients aged from 20 to 69 years after ischemic stroke with Hemiparesis. The patients were divided into 4 groups by a simple randomization. The criteria for this division were: the body side (right or left) affected by paresis and the applied rehabilitation methods. All the patients were applied the recommended kinesitherapeutic method (randomized), 35 therapy sessions, every day for a period of six weeks. Before initiation of therapy and after 6 weeks was measured the total area of the support and path length (COP (Center Of Pressure) measure foot pressure) using stabilometer platform - alpha. The results were statistically analyzed. After treatment studied traits decreased in all groups. The greatest improvement was obtained in groups with NDT-Bobath therapy. NDT-Bobath method for improving the balance of the body is a more effective method of treatment in comparison with of the PNF method. In stroke patients, the effectiveness of NDT-Bobath method does not depend on hand paresis.

The Sensitivity of Endodontic Enterococcus spp. Strains to Geranium Essential Oil.

Enterococci are able to survive endodontic procedures and contribute to the failure of endodontic therapy. Thus, it is essential to identify novel ways of eradicating them from infected root canals. One such approach may be the use of antimicrobials such as plant essential oils. Enterococcal strains were isolated from endodontically treated teeth by standard microbiological methods. Susceptibility to antibiotics was evaluated by the disc-diffusion method. The minimal inhibitory concentration (MIC) of geranium essential oil was investigated by microdilution in 96-well microplates in Mueller Hinton Broth II. Biofilm eradication concentrations were checked in dentin tests. Geranium essential oil inhibited enterococcal strains at concentrations ranging from 1.8-4.5 mg/mL. No correlation was shown between resistance to antibiotics and the MICs of the test antimicrobials. The MICs of the test oil were lower than those found to show cytotoxic effects on the HMEC-1 cell line. Geranium essential oil eradicated enterococcal biofilm at concentrations of 150 mg/mL. Geranium essential oil inhibits the growth of endodontic enterococcal species at lower concentrations than those required to reach IC50 against the HMEC-1 cell line, and is effective against bacteria protected in biofilm at higher concentrations. In addition, bacteria do not develop resistance to essential oils. Hence, geranium essential oil represents a possible alternative to other antimicrobials during endodontic procedures.

The biological activities of cinnamon, geranium and lavender essential oils.

Acinetobacter sp. represent an important cause of nosocomial infections. Their resistance to some antibiotics, their ability to survive on inanimate surfaces in the hospital environment and their ability to produce biofilms contributes to their virulence. The aim of the study was to determine the antibacterial properties of cinnamon, lavender and geranium essential oils against bacteria of the genus Acinetobacter isolated from several clinical materials and from the hospital environment. A comprehensive evaluation of the susceptibility of Acinetobacter sp. clinical strains to recommended antibiotics was performed. The constituents of cinnamon, lavender and geranium essential oils were identified by GC-FID-MS analysis, and their Minimal Inhibitory Concentrations (MICs) against tested clinical strains were determined by the micro-dilution broth method. In addition, the effects of essential oils on the viability of human microvascular endothelial cells (HMEC-1) and glioblastoma cell line (T98G) were evaluated. Cinnamon bark oil was the most active against clinical and environmental strains of Acinetobacter baumannii with MIC values ranging from 0.5 to 2.5 µL/mL. The MIC values for geranium oil were between 7.5 and 9.5 µL/mL, and between 10.5 and 13.0 µL/mL for lavender oil. These essential oils can be best employed in the fight against infections caused by bacteria from Acinetobacter genus as components of formulations for hygiene and disinfection of hospital environment.

[The antibacterial activity of cinnamon oil on the selected gram-positive and gram-negative bacteria]. / Przeciwbakteryjne dzialanie olejku cynamonowego na wybrane bakterie gram-dodatnie i gram-ujemne.

INTRODUCTION: The aim of our study was to determine the antibacterial activity of cinnamon bark oil against Gram-positive and Gram-negative isolates belonging to Staphylococcus, Enterococcus, Enterobacter and Acinetobacter genera come from different clinical specimens. METHODS: The microdilution method was used to determine the minimum inhibitory concentration--MIC for cinnamon bark oil. Susceptibility testing to antibiotics was carried out using disc-diffusion method. RESULTS: Our investigations showed that the tested cinnamon bark oil was inhibiting activity against all isolates. The MIC for Gram-positive bacteria were between 01.25 and 1.5 µl/ml and for Gram-negative between 1.0 and 1.75 µl/ml. The tested bacteria come from Staphylococcus, Enterococcus, Enterobacter and Acinetobacter genera were susceptible to essential oil obtained from Cinnamomum zeylanicum Ness in low concentrations, despite the fact that the bacteria characterized the high resistance to recommended antibiotics. No correlation was found between the antibiotic resistance of the bacterial strains and their sensitivity to essential oil. CONCLUSIONS: The cinnamon bark oil due to the strong activity can be used as alternative antibacterial agents in cosmetics, toiletries and disinfectants applied in hospital environment.

Influence of the surface electrostimulation controlled by muscle contraction on the bioelectric muscle activity and restoration of the hand function in cerebral stroke patients.

BACKGROUND: Impaired limb mobility is the most troublesome condition in patients after cerebral stroke. The application of the electrode-glove might reduce dysfunctions in patients treated due to stroke. OBJECTIVE: The electrode-glove use in rehabilitation is investigated. The study was conducted on 54 patients aged 28 - 72 (the mean age 54.3 ± 10.3) with hemiparesis who had suffered from cerebral stroke. The patients from Group 1 were applied bipolar stimulation of forearm muscles (flexors and extensors of the wrist joint and hand) with the use of two flat electrodes of the same size. With regards to Group 2, in the study used an electrode-glove. METHODS: The excitation threshold was determined once a week with the use of the EMG. SECONDARY OUTCOMES: The evaluation of the electromyographic bioelectric activity of flexor and extensor muscles in the forearm, of the functional mobility of the hand according to the Brunnström test and of the functional mobility of the hand according to the seven-graded task Frenchay scale. RESULTS: The authors of the study observed a much greater improvement in the function of the hand in Group 2, where the mean value increased by 2.1 points. With regards to Group 1, the mean value increased by 1 point and the differences of the results in the studied groups after the therapy were statistically significant (p = 2.830). CONCLUSIONS: The electrical stimulation method controlled/triggered by muscle contractions is an effective method of normalization of muscle tension in the forearm and hand as well as restoration of the hand function in patients with cerebral stroke.

The potential of use basil and rosemary essential oils as effective antibacterial agents.

The considerable therapeutical problems of persistent infections caused by multidrug-resistant bacterial strains constitute a continuing need to find effective antimicrobial agents. The aim of this study was to demonstrate the activities of basil (Ocimum basilicum L.) and rosemary (Rosmarinus officinalis L.) essential oils against multidrug- resistant clinical strains of Escherichia coli. A detailed analysis was performed of the resistance of the drug to the strains and their sensitivity to the tested oils. The antibacterial activity of the oils was tested against standard strain Escherichia coli ATCC 25922 as well as 60 other clinical strains of Escherichia coli. The clinical strains were obtained from patients with infections of the respiratory tract, abdominal cavity, urinary tract, skin and from hospital equipment. The inhibition of microbial growth by both essential oils, presented as MIC values, were determined by agar dilution. Susceptibility testing to antibiotics was carried out using disc diffusion. The results showed that both tested essential oils are active against all of the clinical strains from Escherichia coli including extended-spectrum ß-lactamase positive bacteria, but basil oil possesses a higher ability to inhibit growth. These studies may hasten the application of essential oils in the treatment and prevention of emergent resistant strains in nosocomial infections.

Antimicrobial activity of geranium oil against clinical strains of Staphylococcus aureus.

The aim of this work was to investigate the antibacterial properties of geranium oil obtained from Pelargonium graveolens Ait. (family Geraniaceae), against one standard S. aureus strain ATCC 433000 and seventy clinical S. aureus strains. The agar dilution method was used for assessment of bacterial growth inhibition at various concentrations of geranium oil. Susceptibility testing of the clinical strains to antibiotics was carried out using the disk-diffusion and E-test methods. The results of our experiment showed that the oil from P. graveolens has strong activity against all of the clinical S. aureus isolates-including multidrug resistant strains, MRSA strains and MLS(B)-positive strains-exhibiting MIC values of 0.25-2.50 µL/mL.

Recent patents regarding essential oils and the significance of their constituents in human health and treatment.

Essential oils derived from aromatic plants possess useful properties concerning human health, such as antibacterial, antiviral and antifungal activities. They can be used as natural alternatives to synthetic preparations to prevent and treat infectious diseases caused by multidrug resistant microorganisms. They are used successfully to treat respiratory tract, digestive system and skin infections. Many of them may be applied in anticancer therapy, cardiovascular and nervous system disorders, as well as to reduce the level of cholesterol and decrease and regulate the glucose level. Due to their ability to stimulate adrenal and estrogen hormone production and their antimicrobial properties, they are useful in the treatment of gynecological diseases. Oils are commonly used in the food and cosmetic industry. The present paper describes recent patents concerning potential uses of essential oils in human health and treatment of diseases.

[The antibacterial activity of oregano essential oil (Origanum heracleoticum L.) against clinical strains of Escherichia coli and Pseudomonas aeruginosa]. / Aktywnosc przeciwbakteryjna olejku oreganowego (Origanum heracleoticum L.) wobec szczepów klinicznych Escherichia coli i Pseudomonas aeruginosa.

INTRODUCTION: The aim of this study was to investigate the antibacterial properties of oregano (Origanum heracleoticum L.) essential oil against clinical strains of Escherichia coli and Pseudomonas aeruginosa. The antibacterial activity of oregano essential oil was investigate against 2 tested and 20 clinical bacterial strains of Escherichia coli and 20 clinical strains o Pseudomonas aeruginosa come from patients with different clinical conditions. METHODS: The agar dilution method was used for microbial growth inhibition at various concentrations ofoil. Susceptibility testing to antibiotics was carried out using disc-diffusion method. RESULTS: The results of experiments showed that the tested oil was active against all of the clinical strains from both genus of bacteria, but strains of Escherichia coli were more sensitive to tested oil. Essential oil from Origanum heracleoticum L. inhibited the growth of Escherichia coli and Pseudomonas aeruginosa clinical strains with different patters of resistance. CONCLUSIONS: The obtained outcomes will enable further investigations using oregano essential oil obtained from Origanum heracleoticum L. as alternative antibacterial remedies enhancing healing process in bacterial infections and as an effective means for the prevention of antibiotic-resistant strain development.

The antimicrobial activity of thyme essential oil against multidrug resistant clinical bacterial strains.

AIM: The aim of this work was to investigate the antimicrobial activity of thyme essential oil against clinical multidrug resistant strains of Staphylococcus, Enterococcus, Escherichia, and Pseudomonas genus. MATERIALS: The antibacterial activity of oil was tested against standard strains of bacteria and 120 clinical strains isolated from patients with infections of the oral cavity, abdominal cavity, respiratory and genitourinary tracts, skin, and from the hospital environment. METHODS: Agar diffusion was used to determine the microbial growth inhibition of bacterial growth at various concentrations of oil from Thymus vulgaris. Susceptibility testing to antibiotics was carried out using disk diffusion. RESULTS: Thyme essential oil strongly inhibited the growth of the clinical strains of bacteria tested. CONCLUSIONS: The use of phytopharmaceuticals based on an investigated essential oil from thyme in the prevention and treatment of various human infections may be reasonable.