Biological Active Components Research Articles

Ein Extrakt aus Pelargonium sidoides (EPs® 7630) stimuliert die Phagozytose, den oxidativen Burst und die intrazelluläre Abtötung von Pathogenen in humanen peripheren Blutphagozyten

EXTRACT OF PELARGONIUM SIDOIDES (EPS® 7630) IMPROVES PHAGOCYTOSIS, OXIDATIVE BURST, AND INTRACELLULAR KILLING OF HUMAN PERIPHERAL BLOOD PHAGOCYTES IN VITROClinical data show that EPs ® 7630, an aqueous ethanolic extract from the roots of PELARGONIUM SIDOIDES, can be used for the treatment of upper respiratory tract infections (URTI). The biological effects of the preparation have not been fully investigated. The objective of this study was to examine the impact of EPs ® 7630 on the activity of human peripheral blood phagocytes (PBP). A whole blood-based, flow cytometric assay was used to simultaneously assess phagocytosis and oxidative burst. EPs ® 7630 was applied in therapeutically relevant concentrations between 0 and 30 μg/ml. Compared with controls EPs ® 7630 increased the number of phagocytosing PBP in a concentration-dependent manner, with a maximum enhancement of 56 % at 2 min. The application of EPs ® 7630 also led to a significant increase in the number of burst-active PBP for all time points observed beyond 2 min. Using a microbiological assay, intracellular killing was also enhanced by EPs ® 7630. In conclusion, the positive effects of EPs ® 7630 on phagocytosis, oxidative burst, and intracellular killing of yeast cells as test organisms are important components of the compound's biological activity. Our findings constitute a valuable contribution to understanding the clinical effects of EPs ® 7630.

A hypothetical biological synovial fluid for treatment of temporomandibular joint disease

Temporomandibular Joint disorder (TMD) is a common disorder of mandibular motion system with distinct clinicopathological characteristics. TMD may cause to change in the components of synovial fluid, that affects the functions on lubrication and nutrition of cartilage. Boundary lubrication system contributing to the low friction of joint consists of three parts: lubricin, surface-active phospholipids and hyaluronan (HA). Diminishment of lubrication function is thereby implicated as an adverse contributing factor in degenerative joint diseases such as internal derangement, osteoarthrosis. Moreover, mesenchymal stem cells (MSCs) of synovial membrane can be obtained without irreversible damage, are easily expandable with limited senescence. We postulate that biological active components secreted from MSCs are separated and accumulated by gel permeation chromatography, and then we use the ultra-flirtation of serum and biologically active components to reconstruct the biological synovial fluid in order to rehabilitate the boundary lubrication system and the nutrition of cartilage. Further study investigating the components of biological synovial fluid provides with new treatment strategy for TMD.

Characterization of a heterogalactan: Some nutritional values of the edible mushroom Flammulina velutipes

Production and consume of mushrooms have grown in the world, and beside these, the nutritional properties and biological active components of fungi have received more attention by researchers. Considering these, a mannofucogalactan was isolated from Flammulina velutipes, and characterized using 13C and 1H (obs.), 13C HMQC nuclear magnetic resonance spectroscopy and methylation analysis. The monosaccharide composition of this polymer was determined by GC–MS and showed Fuc p, Man p, and Gal p in the molar ratio 20:16:64, respectively. 13C NMR and 1H (obs.), 13C HMQC indicated an anomeric region containing signals (C-1/H-1) at δ 102.9/5.19, 102.0/5.16, and 98.8/5.05 corresponding, sequentially, to non-reducing end of α- d-Man p, 3- O-substituted α- l-Fuc p, and 6- O- and 2,6-di- O-substituted α- d-Gal p units. Along with methylation analysis, these data showed a structure with a main chain composed of 6- O-substituted Gal p units, partially substituted at O-2 by 3- O- d-mannopyranosyl- l-fucopyranosyl, α- d-mannopyranosyl, and in a minor proportion, α- l-fucopyranosyl groups. Furthermore, some nutritional values of this edible mushroom were evaluated, like amino acid and mineral nutrient contents.

Ganodermataceae: Natural Products and Their Related Pharmacological Functions

The objective of this paper is to review the natural products and the pharmacological functions of Ganodermataceae family. Presently, studies on the bioactive components of Lingzhi are focused on polysaccharides and triterpenes/triterpenoids compounds. New Ganoderma polysaccharides, including their molecular weights, glycosyl residue compositions, glycosyl linkage and branches, are summarized in this paper. Also presented are new types of triterpenes and their characteristics from Lingzhi. Taking Ganoderma lucidum as an example, we reviewed its pharmacological functions in anti-tumor and immune-modulating activities for treating hypoglycemosis, hepatoprotection, and the effect on blood vessel system. Based on the advances in Lingzhi research in the past few decades, both G. lucidum and G. sinense are considered as the representative species of medicinal mushroom Lingzhi in China. Until 2001, G. tsugae was only advised to be used as the materials of the health products. The biologically-active components related to pharmacological functions of these three species were studied more than other Ganodermataceae family species; however, which have been used in less modern folk medicine.

Extract of Pelargonium sidoides (EPs® 7630) improves phagocytosis, oxidative burst, and intracellular killing of human peripheral blood phagocytes in vitro

Clinical data show that EPs 7630, an aqueous ethanolic extract from the roots of Pelargonium sidoides, can be used for the treatment of upper respiratory tract infections (URTI). The biological effects of the preparation have not been fully investigated. The objective of this study was to examine the impact of EPs 7630 on the activity of human peripheral blood phagocytes (PBP). A whole blood-based, flow cytometric assay was used to simultaneously assess phagocytosis and oxidative burst. Calcein-AM stained Candida albicans (DSM 1386) were used as target organisms. Oxidative burst was measured by addition of dihydroethidium (DHE). Target organisms and whole blood were co-incubated and analyzed after 0, 2, 4, 6, 10, and 30 min. Intracellular killing of the target organisms was evaluated by determining the number of surviving yeast cells after co-incubation of C. albicans and human whole blood. EPs 7630 was applied in therapeutically relevant concentrations between 0 and 30 microg/ml. Compared with controls EPs 7630 increased the number of phagocytosing PBP during the observed time points between 2 and 10 min in a concentration-dependent manner, with a maximum enhancement of 56% at 2 min (p=0.002). The application of EPs 7630 also led to a significant increase in the number of burst-active PBP for all time points observed beyond 2 min (p<0.001). The maximum augmentation was 120% after application of 30 microg/ml EPs 7630 at 4 min. Using a microbiological assay, intracellular killing was also enhanced by EPs 7630. This was expressed by a significant reduction in the number of surviving target organisms (p<0.001). The maximum reduction in viable yeast cells (-31%) was observed after co-incubation for 120 min with the highest concentration of EPs 7630 (30 microg/ml). In conclusion, the positive effects of EPs 7630 on phagocytosis, oxidative burst, and intracellular killing of yeast cells as test organisms are important components of the compound's biological activity. Our findings constitute a valuable contribution to understanding the clinical effects of EPs 7630.

The Pharmacological Effects of Odonthobuthus doriae Scorpion Venom and Its Extracted Fractions on Neuro-Muscular Transmission

The effect of Odonthobuthos doriae (O.d) scorpion venom at 0/3, 1 and 3, 10 µg/ml concentrations were investigated on nerve-muscle transmission, using the Twitch tension technique. A concentration of 0.3 µg/ml caused a small change in the twitch height in response to indirect muscle stimulation, but higher concentrations (1, 3, 10 µg/ml) caused a transient augmentation in twitch response followed by a large contracture in the chick biventer cervices (CBC) preparation. This effect could be defined as a complex action of the venom, predominately presynaptic, in which its’ effects on postjunctional synapses is also maintained. In order to find out which bioactive fraction could explain the venom effects, the soluble crude venom was partially separated by the gel filtration method, using a Sephadex G50 column, and four fractions were separated. Two of the four purified fractions (O.d F3, O.d F4) and their LD50 values were lower than the crude venom. Unlike the O.d F1 and O.d F2 fractions, O.d F3 and O.d F4 fractions caused a significant block in the twitch and contracture, in comparison to the control sample . in conclusion, fractions O.d F3 and O.d F4 are supposed to be as the biological active components of the O.d. venom.

Safety evaluation of a triterpenoid-rich extract from bamboo shavings

Triterpenoids, which may have significant application to the development of natural medicines and functional foods as biological active components, are widely distributed throughout the plant kingdom. This paper evaluated the safety of a triterpenoid-rich extract of bamboo shavings (EBS) systematically. (i) Acute toxicity test: The oral maximum tolerated dose of EBS was more than 10 g/kg body weight both in rats and in mice, due to the absence of toxicity according to the criteria of acute toxic classifications. (ii) Mutagenicity test: It had no mutagenicity judged by negative experimental results of Ames test, mouse bone marrow cell micronucleus test and mouse sperm abnormality test. (iii) 30 days feeding study: No abnormal symptoms and clinical signs or deaths had been found in rats in each group during the test. No significant difference had been found in body weight, food consumption and food availability of rats in each test group ( P > 0.05). In addition, no significant differences were found in each hematology value, clinical chemistry value and organ/body weight ratio, either ( P > 0.05). No abnormality of any organ was found during histopathological examination. It can be concluded that the extract of bamboo shavings is of low toxicity and support the use of EBS for various foods.

Cardiovascular protective effects of n-3 polyunsaturated fatty acids with special emphasis on docosahexaenoic acid.

It is widely accepted that n-3 polyunsaturated fatty acids (PUFAs) rich in fish oils protect against several types of cardiovascular diseases such as myocardial infarction, arrhythmia, atherosclerosis, or hypertension. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) may be the active biological components of these effects. Although the precise cellular and molecular mechanisms underlying the beneficial effects are still uncertain, the protective effects of n-3 PUFAs are attributable to their direct effects on vascular smooth muscle cell (VSMC) functions. These n-3 PUFAs activate K(+)(ATP) channels and inhibit certain types of Ca(2+) channels, probably via at least 2 distinct mechanisms. N-3 PUFAs favorably alter the eicosanoid profile and regulate cytokine-induced expression of inducible nitric oxide synthase and cyclooxygenase-2 via mechanisms involving modulation of signaling transduction events. N-3 PUFAs also modulate VSMC proliferation, migration, and apoptosis. These recent data suggest that modulation of these VSMC functions contribute to the beneficial effects of n-3 PUFAs on various cardiovascular disorders. Furthermore, recent studies strongly suggest that DHA has more potent and beneficial effects than EPA. However, many questions about the cellular and molecular mechanisms still remain to be answered.

Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood

Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only β-OH-C14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.

Simultaneous analysis of caffeic acid derivatives and alkamides in roots and extracts of Echinacea purpurea by high-performance liquid chromatography–photodiode array detection–electrospray mass spectrometry

High-performance liquid chromatography (HPLC) coupled with UV photodiode-array detection and electrospray ionization mass spectrometry was developed for the simultaneous analysis of caffeic acid derivatives and alkamides in the roots and extracts of Echinacea purpurea. Caffeic acid derivatives and alkamides produced very abundant peaks in the total ion current chromatogram during negative and positive cone voltage switching. Cichoric acid and the isomer pair, dodeca-2 E,4 E,8 Z,10 E/ Z-tetraenoic acid isobutylamide, were used as a standard for quantification of caffeic acid derivatives and alkamides in E. purpurea. This novel method surpasses previously published ones in product quality control and providing the HPLC chromatographic fingerprints of biological active components in E. purpurea.

Application of Polypyrrole for the Creation of Immunosensors

This review focuses on the use of conducting polymer (CP) films in electrochemical affinity sensors and emphasizes innovative designs and unique applications of immunosensors. The review covers some aspects in the application of polypyrrole (Ppy) for the creation of immunosensors. Polypyrrole film fabrication methods like solvent casting, adsorption, and electropolymerization are presented. The focus, is on electrochemically synthesized Ppy as very promising material for the formation of miniaturized electrochemical immunosensors. Polypyrrole films implemented in various capacities in amperometric, conductometric, and potentiometric immunosensor design are reviewed. The acceptance of immobilization and detection approaches used recently in affinity sensors with critical analysis applied in certain techniques is discussed. The biologically active components (BAC) used for the creation of polypyrrole-based immunosensors are described briefly. Some future trends in the development of polypyrrole-based immunosensors are predicted, as well as possible directions discussed.

Chemopreventive effects of scordinin on diethylnitrosamine and phenobarbital-induced hepatocarcinogenesis in male F344 rats.

Modifying effects of scordinin, a biological active component in garlic, on diethylnitrosamine (DEN)‐ and phenobarbital (PB)‐induced hepatocarcinogenesis were examined in rats. Male F344 rats, 5 weeks old, were divided into 8 groups. After a week, groups 1‐5 were given DEN (100 mg/ kg body weight, i.p.) once a week for 3 weeks, whereas groups 6‐8 received vehicle treatment. Group 2 was given 600 ppm scordinin‐containing diet in the initiation phase. From 4 weeks after the start of experiment, groups 3 and 5 were fed scordinin, and groups 1‐3 and 7 received drinking water containing 500 ppm PB. Group 6 was given scordinin diet alone throughout the experiment (24 weeks). The incidences of hepatocellular adenoma and carcinoma were significantly smaller in group 3 than those in group 1 (P<0.005 and P<0.05, respectively). The average numbers of liver neoplasms in groups 2 and 3 were significantly smaller than in group 1 (P<0.01 and P<0.0001, respectively). Glutathione S‐transferase placental form‐positive foci were also significantly decreased by scordinin treatment in the initiation or promotion phase. Scordinin significantly decreased the mean number of nucleolar organizer regions’ protein (AgNORs)/nucleus in hepatocellular adenoma and carcinoma. AgNORs/nucleus in the non‐lesional area was also significantly decreased by scordinin treatment during the promotion phase. These results suggest that scordinin is a promising chemopreventive agent for liver neoplasia.

Issues and perspectives on the biocompatibility and immunotoxicity evaluation of implanted controlled release systems

Over the past decade, significant advances and discoveries in cell and molecular biology and biomaterials have provided a foundation for the research and development of new, complex controlled release systems. Many of these new controlled release systems utilize active biological components, i.e., proteins and cells, to achieve their intended therapeutic goal. Utilization of bioactive biological materials in implantable controlled release systems has prompted a broad as well as an in-depth interest in the safety and efficacy of these systems. This short review is intended to provide individuals with a perspective on standards and guidance documents which specifically address biological and immunotoxicity response evaluation for safety of controlled release systems from a regulatory point of view.

Effect of docosahexaenoic acid on smooth muscle cell functions

There is increasing evidence that fish oil-enriched diets attenuate the progression of several types of human and experimental renal, intestinal and cardiovascular disorders, including hypertension. Docosahexaenoic acid (DHA), may be one of the active biological component. We previously reported that dietary DHA suppressed the progression of hypertension in stroke-prone spontaneously hypertensive rats (SHRSP). The purpose of this study is to clarify the in vitro effect of DHA on cultured smooth muscle cell functions such as cell growth, hypertrophy, NO release, and intracellular Ca +2 metabolism, which are involved in the regulatory mechanisms of vascular tone. Addition of DHA to the culture medium of aortic smooth muscle cells isolated from SHRSP and normotensive Wistar Kyoto rats (WKY) had no significant effects on cell growth or on cell hypertrophy induced by angiotensin II as measured by flow cytometry. DHA had no stimulatory effect on interleukin-1β (10 mg ml )-induced nitric oxide release from smooth muscle cells of SHRSP, but rather slightly inhibited it. However, the treatment of smooth muscle cells with DHA (30 μM) for 2 days significantly suppressed the increase in intracellular Ca 2+ concentration induced by angiotensin II, but not by thapsigargin. This was due to the suppression of Ca 2+ influx, as determined by Mn 2+ influx experiment. These results indicate that DHA specifically suppresses Ca 2+ mobilization into smooth muscle cells. This may be one of the mechanisms by which dietary DHA prevents the development of hypertension in SHRSP.

Inactivation and toxoiding of biologically-active components of Bordetella pertussis by tea catechins

An ability of tea catechins known as agents for the disinfection to bacteria and viruses were tested on application for toxoiding biologically active components of Bordetella pertussis. The effects on the activities and antigenicity of filamentous hemagglutinin (FHA) and pertussis toxin (PT) were investigated. The activities of FHA and PT were inactivated by catechins at approximately 10(3) times lower dose (0.2 mM) compared with that of formalin. The activity of inactivated FHA was recovered by dialysis against Tris-HCl buffer, pH 8.0, containing glutathione or Tris-HCl buffer, pH 6.0. But the activity of inactivated PT was not recovered. Antigenicity of catechin-treated antigens were investigated by immunization to mice. The sera from mice immunized by catechin-treated FHA or PT were contained antibody against not only catechin-treated but also non-treated FHA or PT. These results suggest that antigenicity of FHA or PT was not destroyed by the treatment with catechin. We prepared pertussis-component vaccines by treatment of several catechins on the condition that FHA or PT activity was not recovered. Higher efficacy were found on the vaccines made by treatment of epicatechin, epicatechin gallate, or epigallocatechin than those by formalin. The vaccine prepared by using epigallocatechin gallate had significant efficacy as well as that by formalin treated one. From these results, it is suggested that tea leaf catechins were effective agents for toxoiding of vaccine components.

Specific modulation of vascular smooth muscle cell functions by docosahexaenoic acid

There are increasing evidences that fish oil-enriched diets attenuate the progression of several types of human and experimental renal, intestinal and cardiovascular disorders including hypertension. Docosahexaenoic acid (DHA) may be one of the active biological component. We previously reported that dietary DHA suppressed the progression of hypertension in stroke-prone spontaneously hypertensive rats (SHRSP). The purpose of this study was to clarify the in vitro effect of DHA on vascular smooth muscle cell functions such as cell growth, hypertrophy, NO release, and intracellular Ca+2 dynamics which involves in the regulatory mechanisms of vascular tone. Addition of DHA to the culture medium of aortic smooth muscle cells isolated from SHRSP and normotensive Wistar Kyoto rats (WKY) had no significant effects on the cell growth, and cell hypertrophy induced by angiotensin II as measured by flow cytometer. DHA did not have a significant effect on interleukin-1 beta (10 ng/ml)-induced nitric oxide release from smooth muscle cells of SHRSP. However, the treatment of smooth muscle cells with DHA (30 microM) for 2 days significantly suppressed the increase in the intracellular Ca2+ concentration induced by 5-hydroxytryptamine, angiotensin II, depolarizing concentration of KCl, but not by thapsigargin. This suppression seems to be due to the suppression of Ca2+ influx, as determined by Mn2+ influx experiment. These results suggest that DHA specifically suppresses receptor-mediated Ca2+ influx in smooth muscle cells. This may be one of the mechanisms by which dietary DHA prevents the development of hypertension in SHRSP.

Effect of serum subfractions from peritoneal dialysis patients on Hep-G2 cell apolipoprotein A-I and B metabolism

We previously showed that uremic serum subfractions isolated from hemodialysis (HD) patients inhibited the production of apolipoprotein (apo) A-I by human hepatoblastoma cells, Hep-G2. Because of the reported differences in atherogenic cardiovascular mortality between HD and peritoneal dialysis (PD) patients, we examined the effect of similar subfractions from PD patients on apo A-I and apo B synthesis. After obtaining informed consent, serum samples from five normal subjects and nine stable PD patients were applied to Sephadex G-25 columns to obtain the serum subfractions used in the various experiments. Sephadex G-25 chromatograms of PD sera showed a broad peak from fractions 30 through 60 (molecular wt 500 to 2000 Da). Control serum showed no peak in this region. PD serum subfractions decreased apo A-I synthesis, secretion, and apo A-I mRNA expression by Hep-G2 cells when compared to subfractions from control subjects. Cholesterol efflux studies showed that conditioned media obtained from Hep-G2 cells incubated with PD serum subfractions inhibited cholesterol efflux from fibroblasts, suggesting a biologically-significant decrease in apo A-I synthesis. PD serum subfractions increased protein synthesis and mRNA expressions of apo B by Hep-G2 cells. Therefore, serum subfractions obtained from PD patients decreased apo A-I and increased apo B synthesis, findings consistent with their serum lipoprotein profiles suggesting that a biologically-active component in these subfractions could contribute to the risk of atherogenic cardiovascular disease in PD.

A Preliminary Study on Strategies for Optimal Fluid-Bed Drying

ABSTRACT The degradation of product quality during convective drying depends on the temperature and water concentration history of the panicles and drying time. For improving product quality in combination with acceptable operation costs, optimal control of the operation variables is required. In this preliminary study the relevance of dynamic optimal operations for batch-wise fluid-bed drying is explored by simulation. Optimal trajectories of operation variables were calculated for a pilot-plant installation by using a model which concerned the drying history of the panicles and the product quality (inactivation of biological active components). The applied objective function was based on an economic criterion combining product quality and operation costs. Although the advances for the chosen pilot-plant application are moderate, in future studies the potentials and relevance of dynamic optimal operations for drying will be quantified for installations on industrial scale.

Pheromone 97*. Der Sexualpheromonkomplex Des Weiblichen Blutbär Thyria Jacobaeae (Lepidoptera, Arctiidae) /Pheromones, 97*. The Sex Pheromone Complex of the Female Arctiid Moth Thyria jacobaeae (Lepidoptera, Arctiidae)

By means of gas chromatographic, mass spectrometric methods and combined GC-electroantennogramm techniques as well as activity comparison with pheromones of related species (3Z,6Z)-c/s-9,10-epoxyeicosadiene 1 and (3Z,6Z)-cis-9,10-epoxyheneicosadiene 2 were identified as the biological active components of the sex pheromone complex of the female arctiid moth Thyria jacobaeae

The role of complement in pregnancy-induced hypertensive disease

Preeclampsia is a pregnancy-induced hypertensive disease with an incidence of about 5% in primigravidas and, being common, it significantly contributes to maternal and neonatal morbidity and mortality. The primary cause remains unknown but might be immunologic, since immunologic aberrations are described in preeclampsia. Activation of the complement system in pregnancy-induced hypertensive disease has been discussed during the last 60 years. It is now strongly indicated that complement activation occurs in preeclampsia. The complement system is very potent and one of the major effector pathways of the process of inflammation. The pathological manifestations, endothelial damage and microvascular injury, and thereby the clinical findings in preeclampsia, may be explained by complement activation, resulting in the direct vascular effects of biological active complement components and complement-mediated activation of leukocytes, with release of potent inflammatory mediators. This new etiological hypothesis might give other options in therapy and prevention of pregnancy-induced hypertensive disease.