Activity of Matricaria chamomilla essential oil against anisakiasis

The increase in diagnosed cases of anisakiasis and the virtual absence of effective treatments have prompted the search for new active compounds against Anisakis L(3) larvae. The biocidal efficacy against different pathogens shown by various essential oils (EO) led us to study the Matricaria chamomilla EO and two of its main components (chamazulene and α-bisabolol) against the L(3) larvae of Anisakis type I. The activity of M. chamomilla EO, chamazulene and α-bisabolol was established by in vitro and in vivo experiments. The EO (125μg/ml) caused the death of all nematodes, which showed cuticle changes and intestinal wall rupture. In the in vivo assays, only 2.2%±1.8 of infected rats treated with M. chamomilla EO showed gastric wall lesions in comparison to 93.3%±3.9 of control. Chamazulene was ineffective, while α-bisabolol showed a high activity to that of the EO in vitro tests but proved less active in vivo. These findings suggest that the larvicidal activity may result from the synergistic action of different compounds of M. chamomilla EO. Neither of the tested products induces irritative damage in the intestinal tissues. In conclusion, M. chamomilla EO is a good candidate for further investigation as a biocidal agent against Anisakis type I.     

Psychopharmacological profile of Chamomile (Matricaria recutita L.) essential oil in mice

In this study, the effect of Matricaria recutita L. essential oil (MEO) on the central nervous system (CNS) of mice was investigated using some behavioral methods. Chemical profiling both by GC and GC-MS analyses of the hydrodistilled essential oil of M. recutita revealed α-bisabolol oxide A (28%), α-bisabolol oxide B (17.1%), (Z)-β-Farnesene (15.9%) and α-bisabolol (6.8%) as the main components. Changes induced by MEO (25, 50 and 100 mg/kg) and reference drug caffeine (25 mg/kg) in spontaneous locomotor activities and motor coordinations of mice were investigated by activity cage measurements and Rota-Rod tests, respectively. Open field, social interaction and elevated plus-maze tests were applied to assess the emotional state of the animals. Further, tail-suspension test was performed for evaluating the effect of MEO on depression levels of mice. As a result, at 50 and 100 mg/kg, MEO significantly increased the numbers of spontaneous locomotor activities, exhibited anxiogenic effect in the open field, elevated plus-maze and social interaction tests and decreased the immobility times of animals in tail suspension tests. The falling latencies in Rota-Rod tests did not change. This activity profile of MEO was similar to the typical psychostimulant caffeine. The exact mechanism of action underlying this stimulant-like effect should be clarified with further detailed studies.     

Drug effect on the rotation thromboelastogram in newborn infants

The impact of the antibiotics carbenicilin and gentamycin on the aggregation and adhesiveness of thrombocytes and on the rotation thrombelastogramme is investigated in healthy newborns. Thrombocyte aggregation was markedly inhibited by carbenicilline; gentamycine had no influence. Thrombocyte adhesiveness was less influenced by both medicaments. The parameters of TEG, viz. reaction time, clotforming time, thrombus elasticity and index were compared before and after rotation and with and without medicaments. Under carbenicilline a prolongation of the clotforming time and a decrease of thrombus elasticity could be shown to exist after rotation. The impact of medicaments on the rotation thrombelastogramme can only be conceived by comparing the values before and after rotation in each case with and without medicament and not simply evaluating it, so that the rotation thrombelastogramms cannot be recommended to be used for assessing the disturbance of thrombocyte function induced by medicaments.     

In vitro propagation and chemical and biological studies of the essential oil of Salvia przewalskii Maxim

The procedure of Salvia przewalskii shoot multiplication and the ability of regenerated plants to produce essential oil is reported. The essential oil was obtained by hydrodistillation from leaves and flowering stems of field-grown plants, and their chemical composition was examined by GC, GC-MS and 1H NMR. The differences in yield as well as qualitative and quantitative composition between the oils isolated from in vitro and in vivo plants were observed. S. przewalskii essential oil was tested for its antimicrobial and cytotoxic properties. It was found that cytotoxicity against human leukemia HL-60 cells and antimicrobial activity (especially, against Staphylococcus aureus and S. epidermidis strains) of oils isolated from in vitro plants were higher than those for oils from in vivo S. przewalskii plants.     

A newly developed assay to study the minimum inhibitory concentration of Satureja spinosa essential oil

AIMS: The minimum inhibitory concentration (MIC) of Satureja spinosa essential oil against Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica, Salmonella serovar Enteritidis PT4 and Bacillus cereus was comparatively assessed with an established optical density method as well as a novel impedimetric method. METHODS AND RESULTS: The impedimetric analysis takes into account information of microbial growth, such as detection time, maximum conductance, and slope of the conductance curve. For each pathogen two levels of inoculation were studied, a high (10(5) CFU ml(-1)) and a low level (10(2) CFU ml(-1)). Non-linear regression analysis was used to fit the data using a modification of a previously published model, from which a more exact value can be obtained for the MIC. Both methods gave similar MICs as shown by t-test statistical analysis. Salm. Enteritidis seems to be the least sensitive to the action of S. spinosa essential oil followed by L. monocytogenes, E. coli, B.cereus and Staph. aureus. The MICs of low inoculum were lower than that of high inoculum. CONCLUSIONS: The new impedimetric assay of MIC of essential oils can be considered a reliable rapid method for screening antimicrobial effectiveness of natural additives. SIGNIFICANCE AND IMPACT OF THE STUDY: Determination of the minimum inhibitory concentration of an essential oil with the simple conductance technique and further study of the mode of action of its components is a good combination for obtaining additional knowledge for industrial application of such natural additives.     

Capability of yeast derivatives to adhere enteropathogenic bacteria and to modulate cells of the innate immune system

Yeast derivatives including yeast cell wall components are promising alternatives to antibiotics with respect to the promotion of health and performance in livestock, based on their capacity to bind enteropathogenic bacteria and to beneficially modulate the immune system. However, these mode(s) of action both in vitro and in vivo are still not well understood. Furthermore, standardization and reproducibility of in vitro techniques (microbiology, cell culture assays) are critical features for the application of yeast derivatives as well as for the proof of effectiveness. Yeast cell wall products are suggested as anti-adhesive agents and are thus proposed to prevent attachment of certain intestinal bacteria by providing alternative adhesion sites to enterobacteria, which contain mannose-specific type I fimbriae such as Escherichia coli or Salmonella spp. and which is well documented. Various in vitro assay techniques have become of paramount importance for biotechnological research since they allow for determination and quantification of potential mode(s) of action. However, in vitro assays may be criticized by product end users as not accurately reflecting in vivo responses. Pro and cons of different assays and their bias will be discussed specifically regarding yeast cell wall components and adhesion of enteropathogenic bacteria. Immunomodulation is a therapeutic approach intervening in auto-regulating processes of the defense system. Yeast derivatives such as beta-glucans are proposed to interact with cells of the innate immune system by receptor recognition. Controversial data in literature and mode(s) of action are reviewed and discussed here.     

Virucidal effect of peppermint oil on the enveloped viruses herpes simplex virus type 1 and type 2 in vitro

The virucidal effect of peppermint oil, the essential oil of Mentha piperita, against herpes simplex virus was examined. The inhibitory activity against herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) was tested in vitro on RC-37 cells using a plaque reduction assay. The 50% inhibitory concentration (IC50) of peppermint oil for herpes simplex virus plaque formation was determined at 0.002% and 0.0008% for HSV-1 and HSV-2, respectively. Peppermint oil exhibited high levels of virucidal activity against HSV-1 and HSV-2 in viral suspension tests. At noncytotoxic concentrations of the oil, plaque formation was significantly reduced by 82% and 92% for HSV-1 and HSV-2, respectively. Higher concentrations of peppermint oil reduced viral titers of both herpesviruses by more than 90%. A clearly time-dependent activity could be demonstrated, after 3 h of incubation of herpes simplex virus with peppermint oil an antiviral activity of about 99% could be demonstrated. In order to determine the mode of antiviral action of the essential oil, peppermint oil was added at different times to the cells or viruses during infection. Both herpesviruses were significantly inhibited when herpes simplex virus was pretreated with the essential oil prior to adsorption. These results indicate that peppermint oil affected the virus before adsorption, but not after penetration into the host cell. Thus this essential oil is capable to exert a direct virucidal effect on HSV. Peppermint oil is also active against an acyclovir resistant strain of HSV-1 (HSV-1-ACV(res)), plaque formation was significantly reduced by 99%. Considering the lipophilic nature of the oil which enables it to penetrate the skin, peppermint oil might be suitable for topical therapeutic use as virucidal agent in recurrent herpes infection.     

Concerning the sensitivity of Acinetobacter calcoaceticus

The strains of Acinetobacter calcoaceticus, var. anitratus (A. c. a.) were isolated in the nosocomial environment as an opportune pathogen. The therapy of choice may be determined after in vitro tests. Our results show following therapeutical possibilities: beta-lactam antibiotics--cephalosporins of IIIrd generation (cefotaxime), also combinations of antimicrobials have shown good results: amoxycillin or ticarcillin with clavulanic acid. Best synergistic effect was found in combination ticarcillin-amikacin.     

A global approach to identify novel broad-spectrum antibacterial targets among proteins of unknown function

Attempted allelic replacement of 144 Streptococcus pneumoniae open reading frames of previously uncharacterized function led to the identification of 36 genes essential for growth under laboratory conditions. Of these, 14 genes (obg, spoIIIJ2, trmU, yacA, yacM, ydiC, ydiE, yjbN, yneS, yphC, ysxC, ytaG, yloI and yxeH4) were also essential in Staphylococcus aureus and Haemophilus influenzae or Escherichia coli, 2 genes (yrrK and ydiB) were only essential in H. influenzae as well as S. pneumoniae and 8 genes were necessary for growth of S.pneumoniae and S. aureus and did not have a homolog in H. influenzae(murD2, ykqC, ylqF, yqeH, ytgP, yybQ) or were not essential in that organism (yqeL, yhcT). The proteins encoded by these genes could represent good targets for novel antibiotics covering different therapeutic profiles. The putative functions of some of these essential proteins, inferred by bioinformatic analysis, are presented. Four mutants, with deletions of loci not essential for in vitro growth, were found to be severely attenuated in a murine respiratory tract infection model, suggesting that not all targets for antibacterial therapeutics are revealed by simple in vitro essentiality testing. The results of our experiments together with those collated from previously reported studies including Bacillus subtilis, E. coli and Mycoplasma sp. demonstrate that gene conservation amongst bacteria does not necessarily indicate that essentiality in one organism can be extrapolated to others. Moreover, this study demonstrates that different experimental procedures can produce apparently contradictory results.     

Synergistic activity of coriander oil and conventional antibiotics against Acinetobacter baumannii

In this study we investigated the existence of synergistic antibacterial effect between coriander (Coriandrum sativum L.) essential oil and six different antibacterial drugs (cefoperazone, chloramphenicol, ciprofloxacin, gentamicin, tetracycline and piperacillin). The antibacterial activity of coriander oil was assessed using microdilution susceptibility testing and synergistic interaction by checkerboard assays. The association of coriander essential oil with chloramphenicol, ciprofloxacin, gentamicin and tetracycline against Acinetobacter baumannii showed in vitro effectiveness, which is an indicator of a possible synergistic interaction against two reference strains of A. baumannii (LMG 1025 and LMG 1041) (FIC index from 0.047 to 0.375). However, when tested the involvement between coriander essential oil and piperacillin or cefoperazone, the isobolograms and FIC index showed an additive interaction. The in vitro interaction could improve the antimicrobial effectiveness of ciprofloxacin, gentamicin and tetracycline and may contribute to resensitize A. baumannii to the action of chloramphenicol.     

Effect of atorvastatin and clopidogrel on cellular immune function

Lipid lowering therapy by statins and antiaggregation have become the basis of any anti-atherosclerotic prophylaxis either as primary or secondary prophylaxis. As several recent papers indicated immunosuppressive properties of statins we investigated changes in lymphocyte subpopulations, apoptosis markers, and cellular immune response towards mitogens after a short-term therapy with atorvastatin and clopidogrel.Nine healthy volunteers (four male, five female, age ranging from 26 to 43 years) were treated with 20 mg atorvastatin for 4 weeks and for 2 additional weeks with 20 mg atorvastatin and 75 mg clopidogrel after oral consent was given. Lymphocyte subpopulations were counted by flow cytometry. To assess cellular in vitro immune function, lymphocyte transformation tests with four mitogens (PHA, ConA, PWM, and OKT3) were performed.Absolute leucocyte counts remained unchanged as well as the granulocyte, monocyte, lymphocyte, and lymphocyte subpopulation counts. There were no detectable changes in markers of cell activation (HLA-DR, CD25, CD69, and CD86) or apoptosis (CD95, annexin). Cellular in vitro responses towards four mitogens did not show significant changes after atorvastatin nor after atorvastatin plus clopidogrel treatment.In conclusion, our data show that atorvastatin is not an immunosuppressive drug under therapeutical conditions.     

Antibacterial and antioxidant activity and essential oil composition of Grammosciadium scabridum Boiss. from Iran

The in vitro antibacterial and antioxidant activity of the essential oil and its two main components of Grammosciadium scabridum Boiss. (Apiaceae) growing wild in Iran, as well as the composition of its essential oil were studied. A total of 19 compounds representing 99.9% of the oil has been identified. Gamma-Terpinene (73.5%), p-cymene (14.2%) and (E)-beta-farnesene (5.3%) were characterized as the main components. The oil showed remarkable activity against three Gram-negative and four Gram-positive test bacteria, with minimal inhibitory concentration (MIC) values ranging from 0.31 to 10.00 mg/ml. The oil and its two main components were also subjected to screening for their possible antioxidant activity by using the 2,2-diphenyl-l-picrylhydrazyl (DPPH) assay. The free radical scavenging capacity of the oil was determined with an IC50 value of 6.6 mg/ml.     

Chemical composition and antitermitic activity against Coptotermes formosanus Shiraki of Cryptomeria japonica leaf essential oil

Cryptomeria japonica D. Don, called 'sugi' in Japanese, is an abundant and renewable potential resource of valuable natural products that may serve as natural biocides in Taiwan. Hydrodistillation (HD) and steam distillation (SD) were used for extracting the essential oils from C. japonica leaves in this study. The chemical constituents of the two leaf essential oils were identified by GC/MS analysis, and their antitermitic activities were evaluated. The results from the antitermitic tests against Coptotermes formosanus Shiraki showed that the leaf essential oils extracted by HD and SD had excellent antitermitic activities, with LD(50) values of 1.57 and 1.72 mg/g after 7 d of testing, respectively. Comparison of the chemical composition of the two leaf essential oils revealed that oxygenated diterpenes (kaur-16-ene) and oxygenated sesquiterpenes (β-elemol) were the dominant constituents. In addition, 13 constituents isolated from the C. japonica leaf essential oils were tested individually against C. formosanus. Among these constituents, β-elemol and α-terpineol achieved 100% termite mortality at the dosage of 1 mg/g after 7 d of testing. These results demonstrated that both C. japonica leaf essential oils as well as β-elemol and α-terpineol possessed commendable antitermitic activity.     

Purification of active matrix metalloproteinase catalytic domains and its use for screening of specific stromelysin-3 inhibitors.

The matrix metalloproteinase (MMP) stromelysin-3 (ST3) has been shown to be involved in malignant tumor progression and therefore represents an attractive therapeutical target. In order to screen for ST3 synthetic inhibitors, we have produced and purified the catalytic domain of ST3, matrilysin, stromelysin-2, and membrane type-1 MMP from inclusion bodies in a bacterial system. Our strategy allowed the purification of MMPs directly in the active form, thereby avoiding in vitro activation. A total of 140,000 synthetic compounds from the Bristol-Myers Pharmaceutical Research Institute chemical deck were tested, using a substrate-based colorimetric enzymatic assay, in which ST3 activity was evaluated through its ability to cleave and inactivate alpha-1 proteinase inhibitor. One ST3 inhibitor belonging to the cephalosporin family of antibiotics was thereby identified.     

儿童泌尿系感染的病原学分析

探讨目前儿童泌尿系感染病原体的变化趋势,为临床治疗提供实验依据。分析2008年1月至2009年10月住院治疗的357例尿细菌培养、支原体体外培养、衣原体检测阳性的泌尿系感染患儿病原体的分布情况。结果显示,尿细菌培养和支原体体外培养、衣原体检测前未应用过抗生素的患儿其阳性率为82.2%,而应用过抗生素的患儿其阳性率为31.8%,两者相比具有统计学意义(P0.01)。在检测的357例阳性标本中,革兰阴性杆菌占74.7%,其中以大肠埃希菌为主,占46.2%;革兰阳性球菌占14.8%,其中肠球菌占10.9%;真菌占3.1%,支原体占4.8%,衣原体占2.5%。临床要密切关注儿童泌尿系感染病原体的分布变迁情况,以便于为临床的诊断和治疗提供可靠的实验依据。     

Anti-inflammatory effects of Melaleuca alternifolia essential oil on human polymorphonuclear neutrophils and monocytes

OBJECTIVE AND DESIGN: The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and anti-oxidative effects remain unclear. In this study, we investigated in vitro the possible role of whole M. alternifolia EO as a modulator of the oxidative response, i.e. reactive oxygen species (ROS) production, of leukocytes (monocytes and polymorphonuclear neutrophils (PMNs)) in humans. METHODS: Whole blood leukocytes from healthy human volunteers (n = 7), isolated from erythrocytes by haemolytic shock, were incubated for 30 min with M. alternifolia EO (0-0.1%) to determine their ROS production by flow cytometry with or without stimulation of cells. We compared the effects of 3 different stimulating agents acting differently on transductional pathways to stimulate the ROS production: a phorbol ester (PMA), formyl-methionyl-leucyl-phenylalanine (fMLP) and opsonised zymosan (OZ). RESULTS: As attested by the Krüskall-Wallis test, M. alternifolia EO at 0.1% directly stimulated ROS production by PMNs (x 8.7 vs. 0% EO, p < 0.05) and increased the intracellular ROS produced by monocytes. Whichever the stimulating agent used (PMA, fMLP or OZ), M. alternifolia EO decreased the intracellular ROS production at the dilution of 0.1% by PMNs and monocytes, more so with PMNs. CONCLUSION: M. alternifolia EO may be both a direct active mediator of the bactericidal action of the circulating leukocytes and may be efficient in protecting the organism from an excess of ROS, through an anti-oxidant and radical scavenging activity.     

Chemical composition and anti-Helicobacter pylori effect of Satureja bachtiarica Bunge essential oil

Resistance of H. pylori strains to common antibiotics has been developed in different parts of the world and continues to increase. It is important to investigate the novel and efficient anti-H. pylori drugs, among which the plants would be suitable sources.Satureja bachtiarica Bunge is traditionally used as antimicrobial agent. In this study, we evaluated the antibacterial activity of S. bachtiarica Bunge essential oil against 10 clinical isolates of Helicobacter pylori by disc diffusion and agar dilution methods. The chemical composition of essential oil was analyzed by GC and GC–MS. Carvacrol (45.5%) and thymol (27.9%) were the primary constituents of oil, followed by p-cymene (4.4%), and γ-terpinene (4.0%). S. bachtiarica essential oil showed strong antibacterial activity against clinical isolates of H. pylori (17.6 ± 1.1 mm and 0.035 ± 0.13 μl/ml). Carvacrol, as the first main component, had a significant role in this effect, whereas in the presence of thymol, the antibacterial effect of carvacrol was reduced. Therefore, S. bachtiarica essential oil can be applied as an alternative agent for treatment of H. pylori infections. More studies would be required to better clarify its mechanism of action on H. pylori.     

Determination of amphetamine,methamphetamine and amphetamine-derived designer drugs or medicaments in blood and urine

This paper reviews procedures for the determination of amphetamine, methamphetamine and amphetamine-derived designer drugs or medicaments in blood and urine. Papers published from 1991 to early 1997 were taken into consideration. Gas chromatographic and liquid chromatographic procedures with different detectors (e.g., mass spectrometer or diode array) were considered as well as the seldom used thin-layer chromatography and capillary electrophoresis. Enantioselective procedures are also discussed. A chapter deals with amphetamine-derived medicaments, e.g. anoretics, antiparkinsonians or vasodilators, which are metabolized to amphetamine or methamphetamine. Differentiation of an intake of such medicaments from amphetamine or methamphetamine intake is discussed. Basic information about the biosample assayed, internal standard, work-up, GC column or LC column and mobile phase, detection mode, reference data and validation data of each procedure is summarized in Tables. Examples of typical applications are presented.     

Praziquantel and albendazole damaging action on in vitro developing Mesocestoides corti (Platyhelminthes: Cestoda)

Parasitic flatworms present several steps of body architecture rearrangement during their fast transition from one developmental stage to another, which are, at least in part, responsible for their evasion from host immune response. Besides, different developmental stages present different degrees of susceptibility to drug action, and the identification of more susceptible stages is of importance for the definition of therapeutical approaches. Mesocestoides corti (syn. Mesocestoides vogae) is considered a good model to study cestode biology because it can be easily manipulated both in vivo and in vitro and due to its relatively close relationship to cestodes of medical relevance, such as those from genera Echinococcus or Taenia. We have analyzed the damaging action of two broad spectrum anthelmintic drugs (praziquantel and albendazole) throughout the in vitro strobilization process of M. corti in order to identify developmental stages or body structures more susceptible to these drugs. Tetrathyridia (larval stage) and segmented-induced worms were cultivated and treated with praziquantel and albendazole. Whole mounted samples, taken from different developmental stages, were fixed and stained with fluorophore-labeled WGA lectin and phalloidin for the analysis of tegument and muscles, respectively. Confocal laser scanning microscopy was used to identify anatomical changes and lesions caused by each anthelmintic drug in a 3D view. We demonstrated that both praziquantel and albendazole cause extensive tissue damage, especially on tegument, and that adult forms were the most susceptible to drug exposure.