NMR and IR analysis of natural substances isolated from Cordyceps medicinal mushrooms.

There exist about 750 species of Cordyceps at present. A high price of natural Cordyceps and its lack in nature caused that the attention has been focused to its cultivation in laboratory conditions. The demand for this “fungus-parasite” is still quite high nowadays, as shown by the amount of commercial nutritional supplements. Phytochemical diversity has ensured that Cordyceps is used as an immunomodulatory and an antioxidant; it has anti-cancer, anti-inflammatory, anti-diabetic, antibacterial, anti-HIV effects. In the present study we focused on NMR and IR analyses of natural substances isolated from two species of Cordyceps: Cordyceps sinensis MFTCCB025/0216, MFTCCB026/0216 and Paecilomyces hepiali MFTCCB023/0216. Two types of rice substrates (Oryza sativa Indica and Oryza sativa Japonica) were used for cultivation. A total of five methanol extracts obtained by a reflux method of the ground mushroom were analysed. To determine the quality and quantity of the major chemical compounds, 1D and 2D NMR analysis has been used with 1H, 13C, COSY, NOESY, HSQC, HMBC and DEPT spectra. IR spectroscopy was chosen as a complementary analysis to determine functional groups. Linoleic acid, oleic acid and mannitol were identified as major compounds of the methanol extracts. Tyrosine, alanine, urea and the others biologically interesting substances were found as minor components.

Antioxidative Protection of Squalene Adjuvant and Rabies Vaccine with Adjuvant.

The authors verified the possibility of antioxidative protection of squalene adjuvant emulsions by the antioxidants α-tocopherol and ß-carotene. They determined the influence of ß-carotene on the stability and antigenic effectiveness of adjuvant emulsion in combination with rabies vaccine. The composition of the adjuvant emulsions or vaccines was: 2.5% squalene; 6% detergents; 0.5% antioxidant; 91% water phase. The oxidative injury after UV-irradiation was followed by the detection of the peroxide value of the emulsions. The stability of the emulsions was evaluated by the determination of the emulsion's particle size. The level of rabies antibodies (RAB) in mice sera until day 90 after vaccination, was determined by the rapid fluorescent focus inhibition test. In the in vitro system of squalene adjuvant, α-tocopherol acted as a prooxidant, while ß-carotene effectively reduced the oxidative injury. The homogenization of the squalene adjuvant during a prolonged period from 8 to 10 min did not change the particle size. The oxidation processes were efficiently reduced by ß-carotene during the preparation process and also during the 70-d storage. The vaccine with ß-carotene induced a gradual increase in the RAB levels with the highest value on day 28. While the inactivated rabies vaccine with adjuvant without ß-carotene developed a rapid formation of RAB, the application of the vaccine with ß-carotene induced a slower but more uniform production of RAB. The level of RAB was significantly higher after the application of the vaccine with ß-carotene and reached the protective value of 0.5 IU/mL, in contrast to the vaccine without ß-carotene.

Antibacterial activity, quality and stability study of creams with new potential silver(I) complexes and in vivo case report.

The aim of this study was to evaluate the antibacterial activity, quality and stability of creams (at 1 % concentration) prepared with synthesized silver(I) complexes: [Ag(Nam)2]NO3·H2O ( AgNam), [Ag2(HGly)2]n(NO3)2n (AgGly) (Nam - nicotin-amide, Gly - glycine) and silver(I) sulfadiazine (AgSD), which is commercially available. Antibacterial activity was evaluated by agar well diffusion method and in in vivo case. The pure silver(I) complexes as well as all three tested creams loaded with AgGly, AgSD and AgNam showed antibacterial potential. Moreover, the creams loaded with AgGly and AgNam showed higher antibacterial effects against S. aureus and B. subtilis than the cream loaded with AgSD. In terms of appearance, all cream samples were opaque and odourless, and no phase separation was observed. Creams were soluble in water (o/w emulsions) and they had a pseudoplastic behaviour. The pH of the creams was in the range of 4.87-5.75. No visible changes were observed in the case of commercially used AgSD cream during one month testing period at conditions -16 ± 1 °C; 6 ± 1 °C and 56 % relative humidity; 20 ± 1 °C and 58 % relative humidity and 40 ± 1 °C and 75 % relative humidity. However, creams containing AgGly and AgNam changed their colour depending on the tested conditions.

Selected aspects of integrated environmental management.

Current trends in environmental care are characterised by the principles of comprehensiveness, integration, interdisciplinarity and transregionality. It is in this spirit that the methodological, legislative and economic instruments develop, and this whole trend is referred to using the term 'environmental management'. Environmental management constitutes a set of technical instruments and methods for managing the environment, based upon an identification of the environmental aspects of products, activities and services of any type of organisation, with the aim to adopt and implement effective proactive measures to reduce their negative impact on the environment. These measures, along with their level, relate to an organisation's possibilities, the market's pressure on it, and primarily with the organisation's management's awareness, maturity and ability to anticipate the introduction of stricter requirements. Environmental management, in the industrial production and service sectors, is one of the the most effective instruments for achieving the priority goal, which is to minimise the negative impact of production activities on the individual elements of the environment as part of the global trend of reducing the negative impact of human activity on the environment. The objective of this article is to indicate possibilities to improve the environment through the basic principles and techniques of integrated environmental management.

Antioxidant Activity and Infrared Spectroscopy Analysis of Alcoholic Extracts Obtained from Paecilomyces hepiali (Ascomycetes).

This research studies the influence of substrate on the antioxidant activity of alcohol extracts of Paecilomyces hepiali. We used corn, rice, millet, and peas as substrates. Antioxidant activity was measured with the DPPH radical scavenging method. Concentrations of extracts (6.25, 3.12, 1.56, 0.78, and 0.39 mg/mL) were applied in all evaluations. Overall antioxidant activity was expressed as the concentration of substrate that decreased DPPH radical levels by 50% (IC50DPPH) for 7 methanol and 7 ethanol extracts. A comparison of IC50DPPH allowed us to conclude that the methanol extracts are more active in scavenging stable DPPH radicals than are the ethanol extracts. The substrate with antioxidant properties most suitable for cultivation of P. hepiali was rice supplemented with non-defatted soy flour. The extract most effective in scavenging stable radicals was the methanol extract of sample 4 (IC50DPPH = 2.33 mg/mL) cultivated on rice with nondefatted soy flour. The methanol extract of sample 7 cultivated on peas was less effective (IC50DPPH = 11.50 mg/mL). By crystallizing these extracts, we managed to obtain sufficient quantities of 6 samples in a solid state, for which infrared spectra were measured and confirmed the presence of amino acids in the extracts.

Free-Radical Scavenging Activities of Cultured Mycelia of Paecilomyces hepiali (Ascomycetes) Extracts and Structural Characterization of Bioactive Components by Nuclear Magnetic Resonance Spectroscopy.

Current research is focused on testing the cultivation of Paecilomyces hepiali mycelia on various plant substrates and producing fungus or mycelial biomass with qualitatively interesting substances. P. hepiali mycelia was cultivated using solid-state fermentation of different substrates. Mycelial biomass was then analyzed, and antioxidant activity was evaluated using the DPPH radical scavenging method for different ethanolic extracts based on a millet substrate (extract 1) or a chickpea substrate (extract 2). Extract 1 corresponds to a half-maximal DPPH radical inhibitory concentration of 1.73 mg/mL; the inhibitory concentration of ethanol extract 2 was almost 4.5 times higher at 7.92 mg/mL. Extracts 1 and 2 were separated into fractions by column chromatography and the chemical structures were determined for the substances that formed the most effective fraction of sample 1. The chemical structures of all compounds in the most active fraction of sample 1 were analyzed by 1H, 13C, distortionless enhancement by polarization transfer, correlation spectroscopy, heteronuclear single-quantum correlation spectroscopy, and heteronuclear multiple-bond correlation spectra.