Analyzing the Carotenoid Composition of Melilot (Melilotus officinalis (L.) Pall.) Extracts and the Effects of Isolated (All-E)-lutein-5,6-epoxide on Primary Sensory Neurons and Macrophages.

Melilotus officinalis is known to contain several types of secondary metabolites. In contrast, the carotenoid composition of this medicinal plant has not been investigated, although it may also contribute to the biological activities of the drug, such as anti-inflammatory effects. Therefore, this study focuses on the isolation and identification of carotenoids from Meliloti herba and on the effect of isolated (all-E)-lutein 5,6-epoxide on primary sensory neurons and macrophages involved in nociception, as well as neurogenic and non-neurogenic inflammatory processes. The composition of the plant extracts was analyzed by high performance liquid chromatography (HPLC). The main carotenoid was isolated by column liquid chromatography (CLC) and identified by MS and NMR. The effect of water-soluble lutein 5,6-epoxide-RAMEB (randomly methylated-ß-cyclodextrin) was investigated on Ca2+-influx in rat primary sensory neurons induced by the activation of the transient receptor potential ankyrin 1 receptor agonist to mustard-oil and on endotoxin-induced IL-1ß release from isolated mouse peritoneal macrophages. (all-E)-Lutein 5,6-epoxide significantly decreased the percent of responsive primary sensory neurons compared to the vehicle-treated stimulated control. Furthermore, endotoxin-evoked IL-1ß release from macrophages was significantly decreased by 100 µM lutein 5,6-epoxide compared to the vehicle-treated control. The water-soluble form of lutein 5,6-epoxide-RAMEB decreases the activation of primary sensory neurons and macrophages, which opens perspectives for its analgesic and anti-inflammatory applications.

[The medical psychology in Hungary: way of thinking, frame of references and applications]. / Az orvosi pszichológia fejlodése hazánkban: szemléleti keretek, határterületek és alkalmazások.

The present study aims at overviewing the different stages of the education of medical psychology in Hungary. In the perspective of the achievements of cognitive and affective neuroscience as well as developmental psychology, we intend to define the roots of doctor-patient relationship, certain determining factors of diagnosis forming and therapy efficiency as well as the essential components of empathy. We attest to the fact that the current psychosomatic medicine with its evidence-based achievements is a valid component and constitutive pillar of everyday medical care. The integrated university level instruction of medical psychology and behavioral sciences aims at consolidating this perspective. We also emphasize that the ill body hinders adequate psychological functioning and mental disorders provide risk factors concerning somatic illnesses. Through spreading health psychology perspective, we support the concept that in everyday medical care, health protection and the development of health conscious behavior relies on responsible, informed and committed decision-making patients as well as on similarly educated extended medical care teams. Orv Hetil. 2018; 159(36): 1455-1464.

A phenotypic in vitro model for the main determinants of human whole heart function.

This article details the construction and testing of a phenotypic assay system that models in vivo cardiac function in a parallel in vitro environment with human stem cell derived cardiomyocytes. The major determinants of human whole-heart function were experimentally modeled by integrating separate 2D cellular systems with BioMicroelectromechanical Systems (BioMEMS) constructs. The model features a serum-free defined medium to enable both acute and chronic evaluation of drugs and toxins. The integration of data from both systems produced biologically relevant predictions of cardiac function in response to varying concentrations of selected drugs. Sotalol, norepinephrine and verapamil were shown to affect the measured parameters according to their specific mechanism of action, in agreement with clinical data. This system is applicable for cardiac side effect assessment, general toxicology, efficacy studies, and evaluation of in vitro cellular disease models in body-on-a-chip systems.

Effects of some natural carotenoids on TRPA1- and TRPV1-induced neurogenic inflammatory processes in vivo in the mouse skin.

Mechanisms of the potent anti-inflammatory actions of carotenoids are unknown. Since carotenoids are incorporated into membranes, they might modulate transient receptor potential ankyrin 1 and vanilloid 1 (TRPA1 and TRPV1) activation predominantly on peptidergic sensory nerves. We therefore investigated the effects of three carotenoids (ß-carotene, lutein and lycopene) on cutaneous neurogenic inflammation. Acute neurogenic edema and inflammatory cell recruitment were induced by smearing the TRPA1 agonist mustard oil (5%) or the TRPV1 activator capsaicin (2.5%) on the mouse ear. Ear thickness was then determined by micrometry, microcirculation by laser Doppler imaging and neutrophil accumulation by histopathology and spectrophotometric determination of myeloperoxidase activity. The effects of lutein on the stimulatory action of the TRPA1 agonist mustard oil were also tested on the guinea-pig small intestine, in isolated organ experiments. Mustard oil evoked 50-55% ear edema and granulocyte influx, as shown by histology and myeloperoxidase activity. Swelling was significantly reduced between 2 and 4 h after administration of lutein or ß-carotene (100 mg/kg subcutane three times during 24 h). Lutein also decreased neutrophil accumulation induced by TRPA1 activation, but did not affect mustard oil-evoked intestinal contraction. Lycopene had no effect on any of these parameters. None of the three carotenoids altered capsaicin-evoked inflammation. It is proposed that the dihydroxycarotenoid lutein selectively inhibits TRPA1 activation and consequent neurogenic inflammation, possibly by modulating lipid rafts.

Composition and antipseudomonal effect of essential oils isolated from different lavender species.

The aims of our research include the phytochemical characterization of the essential oils extracted from different lavender species (Lavandula vera L., L. intermedia L., L. pyrenaica DC., and L. stoechas subsp. stoechas) that are important from therapeutic and economic aspects, as well as the optimization of the tube dilution method for microbiological investigation of the effect of essential oils on the selected Pseudomonas strain. The chemical and percentage compositions of the essential oils were analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The main component of the essential oils in the majority of lavender species was linalool, while L-fencone was identified in the largest amount in L. stoechas subsp. stoechas. Tube dilution is one of the most frequently used methods for microbiological investigation. Its advantages are that it is easy to perform and provides information about minimum bactericidal concentration (MBC). Because of the lipophilic character of essential oils, the tube dilution method should be optimized for the detection of antibacterial activity of these plant extracts, and, therefore, emulsions containing 0.2% polysorbate 80 were prepared from the samples. In each of the eight investigated essential oils an inhibitory effect was detected, and MBC values were in the range 12.5-50 microL/mL in seven cases.

Comparison between ultrasound-guided and digital-guided anesthesia before prostatic biopsy.

OBJECTIVE: Our target in this study is to evaluate the efficacy of ultrasound (US)-guided anesthesia in comparison it to the digital-guided one, considering pain and discomfort reduction, during prostate biopsy. MATERIALS AND METHODS: We analyzed 150 patients that underwent prostate biopsy between March 2011 and January 2012; conditions to enter the sample were: elevated PSA levels and/or psa ratio free/total less than 15% and/or detection of alteration via ultrasound examination and/or a positive outcome of a digital rectal examination. Patients were randomized into two groups. In 75 patients (group A) was performed local US-guided anesthesia with a dose of 10 ml of mepivacaine 1%, in the other 75 patients (group B) a local digital-guided anesthesia was performed, again with an equal dose of 10 ml of mepivacaine 1%. After the biopsy patients were kept under observation for two hours, after that they were asked to provide description of the pain experienced during biopsy, using a 10-point visual analog scale (visual analogue scale; 0 for no pain, 10 for excruciating pain). RESULTS: In group A, 49 patients scored a VAS value of zero, 23 a value of 1 and 2 a value of 3. On the other side, in group B, 9 patients scored a VAS value of 1, 36 a value of 2, 28 a value of 3 and 2 a value of 4. In comparison patients in group A scored VAS values statistically lower than patients in group B (t student test, p < 0.01). CONCLUSIONS: The ultrasound-guided prostatic anesthesia is preferable to the digital-guided, because it considerably reduce the pain related to this procedure.

Patterned cardiomyocytes on microelectrode arrays as a functional, high information content drug screening platform.

Cardiac side effects are one of the major causes of drug candidate failures in preclinical drug development or in clinical trials and are responsible for the retraction of several already marketed therapeutics. Thus, the development of a relatively high-throughput, high information content tool to screen drugs and toxins would be important in the field of cardiac research and drug development. In this study, recordings from commercial multielectrode arrays were combined with surface patterning of cardiac myocyte monolayers to enhance the information content of the method; specifically, to enable the measurement of conduction velocity, refractory period after action potentials and to create a functional re-entry model. Two drugs, 1-Heptanol, a gap junction blocker, and Sparfloxacin, a fluoroquinone antibiotic, were tested in this system. 1-Heptanol administration resulted in a marked reduction in conduction velocity, whereas Sparfloxacin caused rapid, irregular and unsynchronized activity, indicating fibrillation. As shown in these experiments, patterning of cardiac myocyte monolayers solved several inherent problems of multielectrode recordings, increased the temporal resolution of conduction velocity measurements, and made the synchronization of external stimulation with action potential propagation possible for refractory period measurements. This method could be further developed as a cardiac side effect screening platform after combination with human cardiomyocytes.

Carotenoids with anti-Helicobacter pylori activity from Golden delicious apple.

Previously it was reported that hypophasic carotenoids of Golden delicious apple peel showed potent anti-H. pylori activity (MIC(50) = 36 microg/mL), comparable to metronidazole (MIC(50) = 45 microg/mL). To further investigate the involved active carotenoids of the apple peel extracts, seven carotenoids were isolated for the current study: (all-E)-luteoxanthin, (all-E)-neoxanthin, (9'Z)-neoxanthin, (all-E)-antheraxanthin, (all-E)-violaxanthin, (9Z)-violaxanthin and (all-E)-lutein. The MIC(50) values of (all-E)-luteoxanthin, (all-E)-neoxanthin and (9'Z)-neoxanthin were 7.9, 11 and 27 microg/mL, respectively. Other carotenoids and beta,beta-carotene did not exhibit potent anti-H. pylori activity (MIC(50) > 100 microg/mL). An examination of structure and function suggested that active carotenoids contained a monofuranoid ring or an allenic bond in addition to an epoxy group and an additional two or three hydroxyl substituents on the side group.

Pulsed EPR and DFT characterization of radicals produced by photo-oxidation of zeaxanthin and violaxanthin on silica-alumina.

Pulsed electron nuclear double resonance (ENDOR) and two-dimensional (2D)-hyperfine sublevel correlation spectroscopy (HYSCORE) studies in combination with density functional theory (DFT) calculations revealed that photo-oxidation of natural zeaxanthin (ex Lycium halimifolium) and violaxanthin (ex Viola tricolor) on silica-alumina produces the carotenoid radical cations (Car*+) and also the neutral carotenoid radicals (#Car*) as a result of proton loss (indicated by #) from the C4(4') methylene position or one of the methyl groups at position C5(5'), C9(9'), or C13(13'), except for violaxanthin where the epoxide at positions C5(5')-C6(6') raises the energy barrier for proton loss, and the neutral radicals #Car*(4) and #Car*(5) are not observed. DFT calculations predict the largest isotropic beta-methyl proton hyperfine couplings to be 8 to 10 MHz for Car*+, in agreement with previously reported hyperfine couplings for carotenoid pi-conjugated radicals with unpaired spin density delocalized over the whole molecule. Anisotropic alpha-proton hyperfine coupling tensors determined from the HYSCORE analysis were assigned on the basis of DFT calculations with the B3LYP exchange-correlation functional and found to arise not only from the carotenoid radical cation but also from carotenoid neutral radicals, in agreement with the analysis of the pulsed ENDOR data. The formation of the neutral radical of zeaxanthin should provide another effective nonphotochemical quencher of the excited state of chlorophyll for photoprotection in the presence of excess light.

[Medicinal plant research and phytochemical traditions of Pécs]. / Pécsi gyógynövenykutatási és fitokémiai hagyományok.

The authors have summarized the local history of pharmacobotany, phytochemistry and pharmacognosy remembering Szabolcs Nyiredy, the honorary professor of Pécs University.

Status of complementary and alternative medicine in European medical schools.

BACKGROUND: The aim of this study was to investigate whether Complementary and Alternative Medicine (CAM) education is part of the regular medical curriculum in European universities and whether it correlates with the public demand for CAM methods. MATERIALS AND METHODS: Using the World Directory of Medical Schools, 265 medical faculties in EU countries were contacted via e-mail or regular post and asked to fill out a questionnaire of 7 questions concerning CAM education in their establishments. RESULTS: Only 40% of the responding universities were offering some form of CAM training. We could not show any correlation between the public demand for CAM methods and the availability of CAM training in medical universities. CONCLUSIONS: The growing popularity of CAM makes it very important for physicians to gain information on these methods and techniques, thus better incorporation of CAM education into the regular medical curriculum is needed in all European medical faculties.

Inhibition of multidrug resistance of cancer cells by natural diterpenes, triterpenes and carotenoids.

The multidrug resistance (MDR) proteins are member of the ATP-binding cassette superfamily and are present in a majority of human tumors. Their activity is a crucial factor leading to therapeutic failure. It is likely that compounds which inhibit the function of the MDR-efflux proteins such as MDR1 will improve the cytotoxic action of anticancer chemotherapy. Therefore, a search for MDR reversing compounds was conducted among three classes of plant derived compounds such as diterpenes, triterpenes and carotenoids in a hope to find inhibitors without adverse effects in these natural compounds. The inhibition of efflux activity was determined by measuring the accumulation of substrate analogues such as rhodamine in tumor cells in the presence of potential inhibitors. Thus we determined the effect of structurally unrelated diterpenes, triterpenes and carotenoids on reversal of multidrug resistance in MDR-1 gene-transfected L1210 mouse lymphoma cells and MDR mediated multidrug resistance of human breast cancer cells MDA-MB-231 (HTB-26) and MCF-7. The majority of diterpenes, cycloartane triterpenes and carotenoids isolated from vegetables and medicinal plants were able to enhance rhodamine 123 accumulations of MDR-cells. Synergistic interaction was found between epirubicine and resistance modifier terpenoids in vitro. It is supposed that these MDR modulators bind into transmembrane domains and the action of ABC transporters is inhibited by induced conformational changes.

Biological activity of carotenoids in red paprika, Valencia orange and Golden delicious apple.

Carotenoid fractions were extracted from red paprika, Valencia orange peel and the peel of Golden delicious apple. Thus, hypophasic carotenoids of paprika (PM1), orange (PM3) and apple (PM4), and epiphasic extractions of paprika (PM2) and apple (PM5) were obtained by extraction, saponification and partition between MeOH-H(2)O (9:1) (hypophasic) and hexane (epiphasic). A high content of capsanthin was quantified in hypophasic carotenoids (PM1) from red spice paprika, whereas the hypophasic fractions from orange (PM3) and apple (PM4) were mainly composed of violaxanthin, zeaxanthin and lutein. On the other hand, a high content of beta,beta-carotene and beta-cryptoxanthin was found in epiphasic fractions (PM2 and PM5). The extracts were studied for their anti-Helicobacter pylori (H. pylori), anti-human immunodeficiency virus (HIV), cytotoxic, multidrug resistance (MDR) reversal and radical scavenging activity. Among five PM extracts and beta,betacarotene, PM4 showed potent anti-H. pylori activity (MIC(50) = 36 microg/mL), comparable to metronidazole (MIC(50) = 45 microg/mL). The extracts were inactive against HIV. PM3 and PM4 showed slightly higher cytotoxic activity against three human tumor cell lines (squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG) and human promyelocytic leukemic HL-60 cells than against three normal human oral cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF), suggesting a tumor-specific cytotoxic activity. PM1, PM3 and PM4 displayed much higher MDR-reversing activity than (+/-)-verapamil. ESR spectroscopy demonstrated that PM1-5 and beta,beta-carotene produced little or no detectable radical under alkaline conditions and did not scavenge the O(2) (-) produced by the hypoxanthine and xanthine oxidase reaction. On the other hand, PM1 and PM2 scavenged efficiently 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, whereas singlet oxygen was also quenched efficiently by PM5 and PM2. The data suggest the potential importance of carotenoids as possible anti-H. pylori and MDR reversal agents. The active principles in the carotenoid extract might differ, depending upon the types of fruits and vegetables.

Modulation of multidrug resistance and apoptosis of cancer cells by selected carotenoids.

The multidrug resistance (MDR) proteins that belong to the ATP-binding casette superfamily are present in a majority of human tumors and are an important final cause of therapeutic failure. Therefore, compounds which inhibit the function of the MDR-efflux proteins may improve the cytotoxic action of anticancer chemotherapy. The effects of carotenoids were studied on the activity of the MDR-1 gene-encoded efflux pump system. The carotenoids, isolated from paprika and other vegetables, were tested on the rhodamine 123 accumulation of human MDR-1 gene-transfected L1210 mouse lymphoma cells and human breast cancer cells MDA-MB-231 (HTB-26). Capsanthin and capsorubin enhanced the rhodamine 123 accumulation 30-fold relative to nontreated lymphoma cells. Lycopene, lutein, antheraxanthin and violaxanthin had moderate effects, while alfa- and beta-carotene had no effect on the reversal of MDR in the tumor cells. Apoptosis was induced in human MDR1 transfected mouse lymphoma cells and human breast cancer MDA-MB-231 (HTB-26) cell lines in the presence of lycopene, zeaxanthin and capsanthin. The data suggest the potential of carotenoids as possible resistance modifiers in cancer chemotherapy.

[Biomechanical experiments for measuring traction lengthening of the lumbar spine during weight bath therapy]. / A lumbalis gerinc nyúlásának meghatározása súlyfürdóben biomechanikai kísérletek alapján.

INTRODUCTION: Weight-bath as an effective traction therapy has successfully been applied in Hungary for nearly a half century, however, it has still been constrained exclusively to empirical bases until the numerical biomechanical analysis of Bene and Kurutz appeared in 1993. Due to their calculations, for cervical suspension in traction bath, the value and distribution of tensile force along the spine became known, however, the most important information, namely, the deformation of spine segments to be stretched by the therapy is so far unknown. AIMS: In this paper the results of a wide-ranging in vivo biomechanical experimental analysis are presented, aimed partly to obtain the traction deformation effects of weight-bath therapy, partly to obtain the biomechanical traction model of human lumbar segments. The analysis aimed to clear the effect of decompression, extra weights, sex, aging, body height and weight. PATIENTS/METHODS: The experiments have been executed during the prescribed 20 minutes long weight-bath treatment of patients having indication for cervical suspension. Patients with other forms of suspension, like armpit bars, have been excluded from the analysis. Two groups of patients have been distinguished: 67 patients without and 88 patients with extra weight loads of 20-20 N (2-2 kgf) applied on the ankles. Extension values of lumbar segments LIII-IV, LIV-V and LV-SI have been measured. Tensile deformations have been specified as the change of the distance between two spinous process of vertebrae, measured by a special subequal ultrasound method developed by the research group. The experimental results have been evaluated by using special software for analyzing ultrasound pictures. More than 3500 ultrasound pictures of 400 lumbar segments of 155 patients have been measured and evaluated. RESULTS: As for the results, at the end of the treatment, elongation of lumbar segments has been demonstrated practically in 60% of patients without and in 75% of patients with extra weights. The mean extension of a lower lumbar segment after a 20 minutes long weight-bath treatment is about 1.2-1.4 mm, while with extra weights it is about 1.3-1.6 mm, in the average of the deformed segments only. In the average of all segments, the above mean extensions are 0.7-0.9 mm, and with extra weights yield 0.8-1.4 mm. CONCLUSIONS: Just being suspended in water, due to the decompression, even without any extra weights, significant extension has been registered: a mean value of 1.0-1.2 mm per segments for the deformable, and 0.4-0.6 mm for all segments. The 20 minutes long treatment time in itself has a significant effect: the average additional extension is 0.2-0.4 mm per segments without, and about 0.5 mm with extra weights. The deformation capacity of lumbar segments decreases with increasing age of patients. Elongations increase with increasing body height. Body weight shows different tendency depending on the sudden elastic and time-dependent viscous deformations. Significant difference has been observed in reaction time of male and female patients: female patients react later, however the final traction effect seems to be equal.