Differentiation of species of the Baccharis genus by HPLC and chemometric methods.

Phytochemical investigation of the aerial parts of three Baccharis species (Asteraceae family) was performed using HPLC and chemometric methods, with the objective of distinguishing between three morphologically very similar species: Baccharis genistelloides Persoon var. trimera (Less.) DC, B. milleflora (Less.) DC and B. articulata (Lam.) Persoon. With the help of Principal Component Analysis (PCA) and variance weights, it was possible to characterize the chromatographic profiles of the alcoholic extracts of the three species. Application of Soft Independent Modeling of Class Analogy (SIMCA) and K-Nearest Neighbor (KNN) methods on a training set of 74 extracts resulted in models that correctly classified all eight samples in an independent test set.     

Numerical taxonomy characterization of Baccharis genus species by ultraviolet-visible spectrophotometry.

Numerical taxonomy characterization of Baccharis genus species was performed using ultraviolet-visible spectrophotometry. The aim was to present a more convenient, more practical, more economic and faster method based on chemometric methods and UV-vis absorbance to give the most information about species identity and discrimination, especially when their classification has been doubtful. Three Baccharis species: B. genistelloides Persoon var. trimera (Less.) DC, B. milleflora (Less.) DC, and B. articulata (Lam.) Persoon were included in the study. With the help of principal component analysis (PCA) and cluster analysis (CA), we could characterize the three species. Application of soft independent modeling of class analogy (SIMCA) and K-nearest neighbor (KNN) methods on a training set of 65 extracts resulted in models that correctly classified all samples of an independent validation set, eight samples of B. genistelloides Persoon var. trimera (Less.) DC and one sample donated by Prof. Alarich Schultz Herbarium, Porto Alegre-RS, Brazil.     

Short and long-term effects of Baccharis articulata on glucose homeostasis

In this study, the in vivo effect of the crude extract and n-butanol and aqueous residual fractions of Baccharis articulata (Lam.) Pers. on serum glucose levels, insulin secretion and liver and muscle glycogen content, as well as in vitro action on serum intestinal disaccharidase activity and albumin glycation were investigated. Oral administration of the extract and fractions reduced glycemia in hyperglycemic rats. Additionally, the n-butanol fraction, which has high flavonoids content, stimulated insulin secretion, exhibiting an insulinogenic index similar to that of glipizide. Also, the n-butanol fraction treatment significantly increased glycogen content in both liver and muscle tissue. In vitro incubation with the crude extract and n-butanol and aqueous residual fractions inhibited maltase activity and the formation of advanced glycation end-products (AGEs). Thus, the results demonstrated that B. articulata exhibits a significant antihyperglycemic and insulin-secretagogue role. These effects on the regulation of glucose homeostasis observed for B. articulata indicate potential anti-diabetic properties.     

Direct evidence for the plant origin of Brazilian propolis by the observation of honeybee behavior and phytochemical analysis

To identify the plant origin of Brazilian propolis directly, we observed the behavior of honeybees in Minas Gerais State of Brazil. Honeybee workers bit and chewed leaves of the plant, Baccharis dracunculifolia, packed the material into their pollen basket, brought it back to their nest, and used it as propolis. We collected the leaves of B. dracunculifolia and propolis, and compared their constituents by liquid chromatography-mass spectrometry (LC/MS) analysis. There was no difference between the chemical constituents of the ethanol extracts of B. dracunculifolia and those of propolis. This indicates directly that the plant origin of Brazilian propolis is B. dracunculifolia.     

Phenolic enriched extract of Baccharis trimera presents anti-inflammatory and antioxidant activities

Baccharis trimera is a plant popularly used as a tea and to treat gastrointestinal diseases and inflammatory processes as well. The total phenolic content was determined and the antioxidant and anti-inflammatory activities of six extracts (dichloromethane, ethyl acetate, butanol, aqueous, saponin and phenolic) from B. trimera were evaluated. Using carrageenan-induced pleurisy as a model of acute inflammation, the phenolic extract at 15 mg/kg decreased significantly the analyzed parameters when compared to the carrageenan group ( p < 0.05), thus showing potential anti-inflammatory activity. The total phenolic content and antioxidant activity were evaluated by the Folin-Ciocalteau and DPPH methods, respectively. Phenolic and ethyl acetate extracts presented higher antioxidant activity ( p < 0.05) than ascorbic acid. The phenolic extract also showed the highest antioxidant potential in relation to the other extracts, thus suggesting that the antioxidant and anti-inflammatory activities were due to the presence of phenolic compounds.     

Antimicrobial activity of three baccharis species used in the traditional medicine of Northern Chile

The antimicrobial activities of aqueous ethanol and chloroform extracts of three Baccharis species currently used in Northern Chile folk medicine for the treatment of several infectious and inflammatory disorders were tested against Gram-positive and negative bacteria and fungal spp. using the agar-disc diffusion assay. The results indicated that the activity was more pronounced against Gram-positive than against Gram-negative bacteria and yeast. No significant differences on the antibacterial activity were observed in the aqueous ethanol versus chloroform extracts. None of the plant extracts evaluated exhibited any activity against ten fungi tested.     

Kinetic, equilibrium and thermodynamic studies of cadmium (II) adsorption by modified agricultural wastes

Agricultural wastes have great potential for the removal of heavy metal ions from aqueous solution. The contamination of water by toxic heavy metals is a worldwide environmental problem. Unlike organic pollutants, the majority of which are susceptible to biological degradation, heavy metals do not degrade into harmless end products. Discharges containing cadmium, in particular, are strictly controlled because of the highly toxic nature of this element and its tendency to accumulate in the tissues of living organisms. This work aims to develop inexpensive, highly available, effective metal ion adsorbents from natural wastes as alternatives to existing commercial adsorbents. In particular, Tamrix articulata wastes were modified chemically by esterification with maleic acid to yield a carboxyl-rich adsorbent. The adsorption behavior of treated Tamrix articulata wastes toward cadmium ions in aqueous solutions in a batch system has been studied as a function of equilibration time, adsorbent dose, temperature and pH. Results showed that the maximum adsorption capacity was 195.5 mg/g in a pH 4 solution at 30 °C with a contact time of 120 min, an initial concentration of 400 mg/L and an adsorbent dose of 0.3 g/L. The kinetic data were analyzed using pseudo-first-order and pseudo-second-order kinetic models. It was shown that the adsorption of cadmium could be described by a pseudo-second-order equation. The experimental data were also analyzed using the Langmuir and Freundlich models of adsorption. Thermodynamic parameters such as ΔG°, ΔH° and ΔS° have been evaluated and it has been found that the sorption process was spontaneous and exothermic in nature. From all of our data, we conclude that the treated Tamrix articulata wastes investigated in this study showed good potential for cadmium removal from aqueous solutions.     

Chemical analysis and antiviral activity evaluation of Baccharis anomala

The chemical composition and antiviral activity of aqueous extract from Baccharis anomala was studied by bioactivity-guided fractionation. Ethanol precipitation and fractionation by molecular permeation allowed the separation of the anti-herpes simplex virus 1 (HSV-1) active fraction from aqueous extract (Fraction B). Natural Product Reagent A, FeCl₃ and thin-layer chromatography indicated the presence of phenolic compounds in the aqueous extract. Fraction B showed pronounced antiviral activity when tested with HSV-1 strains VR733/ATCC and Acyclovir-resistant 29-R, displaying virucidal but not virustatic activity.     

Unprecedented occurrence of isoaspartic acid in a plant cyclopeptide

Three structurally related cycloheptapeptides, cyclocitropsides A-C, have been isolated from a MeOH extract of the root bark of Citropsis articulata, a medicinal plant in Uganda. Their sequences were elucidated on the basis of their MS/MS fragmentation, extensive 2D-NMR, chemical degradation, and biochemical modifications. Surprisingly, the sequence of cyclocitropside C differed from that of cyclocitropside B only by an Asp(5)/isoAsp(5) substitution. This is the first report of an isoAsp residue in a plant cyclic peptide.     

Impact of gas chromatography and mass spectrometry combined with gas chromatography and olfactometry for the sex differentiation of Baccharis articulata by the analysis of volatile compounds

The Baccharis genus has more than 400 species of aromatic plants. However, only approximately 50 species have been studied in oil composition to date. From these studies, very few take into consideration differences between male and female plants, which is a significant and distinctive factor in Baccharis in the Asteraceae family. Baccharis articulata is a common shrub that grows wild in south Brazil, northern and central Argentina, Bolivia, Paraguay and Uruguay. It is considered to be a medicinal plant and is employed in traditional medicine. We report B. articulata male and female volatile composition obtained by simultaneous distillation–extraction technique and analyzed by gas chromatography with mass spectrometry. Also, an assessment of aromatic differences between volatile extracts was evaluated by gas chromatography with olfactometry. The results show a very similar chemical composition between male and female extracts, with a high proportion of terpene compounds of which β‐pinene, limonene and germacrene D are the main components. Despite the chemical similarity, great differences in aromatic profile were found: male plant samples exhibited the strongest odorants in number and intensity of aromatic attributes. These differences explain field observations which indicate differences between male and female flower aroma, and might be of ecological significance in the attraction of pollinating insects.     

Specific differentiation in Vicia genus by means of capillary electrophoresis

Capillary electrophoresis has been applied to the discrimination of 13 Vicia species, belonging to four sections of Vicia genus. The studied species necessitate of plant growing tests or DNA molecular markers to be distinguished being their seeds morphologically very similar. Alcoholic/saline extracts from dry cotyledons were separated in uncoated fused-silica capillary with iminodiacetic acid isolectric buffer containing hydroxypropylmethylcellulose and acetonitrile. The low intra-specific variation observed for 11 species, suggests that this approach is suitable to carry out species discrimination. Species-specific peaks were identified for V. articulata, V. atropurpurea, V. bithynica, V. benghalensis, V. disperma, V. ervilia, V. monantha, V. sativa and V. villosa. Conversely, V. lutea, V. melanops and V. peregrina, showing very similar electrophoregrams, require other methodological approaches to be discriminated. The discussed CE method appears to have a potential to be regarded as an alternative tool to identify some Vicia species being far less expensive and time consuming than plant growing tests and DNA molecular markers.     

UV-ENHANCED REACTIVATION IN MAMMALIAN CELLS: INCREASE BY CAFFEINE

Abstract— It has been reported that caffeine decreases UV-enhanced reactivation of UV-irradiated Herpes simplex virus in CV-1 monkey kidney cells. That occurred when there was no delay between cell irradiation and virus infection. In the present study, virus infection was delayed following cell irradiation to allow an 'induction'period separate from the 'expression'period which occurs during the virus infection. Thus, the effects of caffeine on 'induction'and 'expression'could be determined separately. Caffeine increased the expression of UV-enhanced reactivation, while causing a small decrease in the 'induction'of enhanced reactivation.     

Physical identification of a virus in a crude leaf extract by its Ferguson plot in agarose gel electrophoresis

Crude extracts of turnip crinkle virus upon agarose gel electrophoresis yield (i) virus patterns unperturbed by contaminants; (ii) plots of mobility vs. gel concentration (Ferguson plots) parallel with those of the purified virus. The parallelism suggests similarity in size and shape but a lower net charge for the crude virus. This result is obtained when gel electrophoresis is carried out either in a continuous buffer or in a discontinuous (moving boundary electrophoresis) buffer system. The latter mode has the substantial benefit of electrophoretic (auto-)concentration of dilute virus sample prior to resolution. Thus, the Ferguson plot analysis in a discontinuous buffer system of turnip crinkle virus can be viewed as a model procedure for the physical identification of other viruses contained in dilute extracts, feasible even in the absence of a prior knowledge as to the nature of, or isolation of, the virus.     

Determination of ginkgolide A,B, and bilobalide in<Emphasis Type="Italic"> biloba</Emphasis> L. extracts by microdialysis-HPLC

A method has been developed for measuring ginkgolide A, B, and bilobalide in ginkgo biloba L. extracts by using a self-assembled microdialysis device coupled on-line to high-performance liquid chromatography. The optimal conditions for dialysis efficiency and chromatographic analysis were investigated. The results show that the dialysis efficiencies for ginkgolide A, B, and bilobalide are 97.8–100.4%. For the spike concentration of 45 mg L^–1, the recoveries of ginkgolide A, B, and bilobalide in ginkgo biloba L. extracts were at 97–100.7% with 3.1–4.7% RSD. The proposed method has therefore been shown to be applicable to the analysis of ginkgolide A, B, and bilobalide in ginkgo biloba L. extracts.     

High-performance liquid chromatographic method for identification and quantification of two isomeric coumarinolignoids-cleomiscosin A and cleomiscosin B-in extracts of Cleome viscosa

A simple, accurate and reproducible reverse-phase HPLC method has been developed for identification and quantification of two isomeric coumarinolignoids, cleomiscosin A and B in different extracts of the seeds of Cleome viscosa using photodiode array detection at 326 nm. Cleomiscosin A and B were separated on a Waters symmetry C(18) column (250 x 4.6 mm with 5.0 microm particle size) with an isocratic elution system composed of acetonitrile-methanol (1:2, v/v) and acetic acid-water (0.5:99.5, v/v) in the ratio of 40:60 (v/v). The calibration curves were linear (r(2) > 0.997) in the concentration ranges of 20-100 microg/mL for both compounds. The limits of detection and quantification were 15 and 20 microg/mL for both cleomiscosin A and B. The intra- and inter-day precisions were 3.68 and 2.22% for cleomiscosin A and 4.22 and 5.06% for cleomiscosin B. The recoveries measured at two different concentration levels varied from 98.03 to 110.06%. The method was used to identify and quantify cleomiscosins A and B in different extracts of Cleome viscosa seeds.     

Antimicrobial activity and phytochemical analysis of crude extracts and essential oils from medicinal plants

We aimed to establish a phytochemical analysis of the crude extracts and performed GC-MS of the essential oils (EOs) of Eugenia uniflora L. (Myrtaceae) and Asteraceae species Baccharis dracunculifolia DC, Matricaria chamomilla L. and Vernonia polyanthes Less, as well as determining their antimicrobial activity. Establishment of the minimal inhibitory concentrations of the crude extracts and EOs against 16 Staphylococcus aureus and 16 Escherichia coli strains from human specimens was carried out using the dilution method in Mueller-Hinton agar. Some phenolic compounds with antimicrobial properties were established, and all EOs had a higher antimicrobial activity than the extracts. Matricaria chamomilla extract and E. uniflora EO were efficient against S. aureus strains, while E. uniflora and V. polyanthes extracts and V. polyanthes EO showed the best antimicrobial activity against E. coli strains. Staphylococcus aureus strains were more susceptible to the tested plant products than E. coli, but all natural products promoted antimicrobial growth inhibition.     

Characterization of isomeric cationic porphyrins with β-pyrrolic substituents by electrospray mass spectrometry: The singular behavior of a potential virus photoinactivator

Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS/MS) have been used to differentiate the 2- and 4-methylpyridyl isomers of free-base and metallated cationic beta-vinylpyridylporphyrins. The analysis by ESI-MS/MS of the deuterated analogs and semiempirical calculations of structural and electronic parameters were also undertaken. The two free-base isomers are easily differentiated by ESI-MS/MS but the presence of a metallic center renders differentiation of the metallated isomers less effective. The data acquired show that of all the studied compounds, the free-base 2-methylpyridyl isomer, which was operative in the in vitro photoinactivation of Herpes simples virus, has a different gas-phase behavior. Local distortion of the macrocycle due to the presence of the beta-vinylpyridyl substituent occurs for all the compounds, but a different electron density distribution can account for the observed gas-phase behavior of this potential virus photoinactivator.     

Novel In Vitro Antioxidant and Photoprotection Capacity of Plants from High Altitude Ecosystems of Colombia

Currently, plants have gained widespread interest as a source of natural sunscreen. Specifically, plants from high altitude ecosystems are exposed to high UVR levels; therefore, they must produce an adaptive chemical response. The aim of this study was to evaluate the photo‐protection and antioxidant capacity in vitro of nine plants from high altitude ecosystems in Antioquia, Colombia (Sphagnum meridense, Calamagrostis effusa, Lycopodiella alopecuroides, Morella parvifolia, Baccharis antioquensis, Pentacalia pulchella, Castilleja fissifolia, Hesperomeles ferruginea and Hypericum juniperinum). B. antioquensis and P. pulchella extracts showed the best results over a broad spectrum UVA‐UVB with antioxidant capacity in vitro. However, B. antioquensis extracts presented the highest absorption coefficient in UVB‐UVA range among plants under study. Furthermore, the gel formulation containing the crude extract of B. antioquensis showed significant values of UVAPF, UVA/UVB ratio, critical wavelength (λ_c) and SPF (3, 0.78 380 nm and 4.73 ± 0.26; respectively), indicating interesting photostability and antiradical capacities. All of these properties could be improve in order to satisfy the requirements for broad‐spectrum UVB/UVA protection. Finally, P. pulchella and B. antioquensis extracts could be a potential source of a new natural sunscreen compounds with photostable and antiradical properties.     

Identification of natural dyes in archeological Coptic textiles by liquid chromatography with diode array detection

Reversed-phase HPLC with diode-array UV-Vis spectrophotometric detection has been used for identification of natural dyes in extracts from wool and silk fibres from archeological textiles. The examined objects originate from 4th to 12th Century Egypt and belong to the collection of Early Christian Art of the National Museum in Warsaw. Extraction from fibres was carried out with HCl solution containing ethanol or with warm pyridine. As the main individual chemical components of natural dyes, anthraquinone, indigoid and flavonoid dyes including alizarin, purpurin, luteolin, apigenin, carminic acid, ellagic acid, gallic acid, laccaic acids A and B and indigotin were found. For pyridine extracts another mobile phase with an optimized gradient of organic modifier concentration was used. With such an eluent the appearance of double peaks for indigotin and indirubin was eliminated. For acidic extraction of dyes from fibres, ethanol was used. Due to its higher boiling point than methanol it evaporates slower from the extraction solution enabling a more efficient extraction of dyes.     

High levels of soluble herpes virus entry mediator in sera of patients with allergic and autoimmune diseases

Herpes virus entry mediator (HVEM) is a newly discovered member of the tumor necrosis factor receptor (TNFR) superfamily that has a role in herpes simplex virus entry, in T cell activation and in tumor immunity. We generated mAb against HVEM and detected soluble HVEM (SHVEM) in the sera of patients with various autoimmune diseases. HVEM was constitutively expressed on CD4(+) and CD8(+) T cells, CD19(+) B cells, CD14(+) monocytes, neutrophils and dendritic cells. In three-way MLR, mAb 122 and 139 were agonists and mAb 108 had blocking activity. An ELISA was developed to detect sHVEM in patient sera. sHVEM levels were elevated in sera of patients with allergic asthma, atopic dermatitis and rheumatoid arthritis. The mAbs discussed here may be useful for studies of the role of HVEM in immune responses. Detection of soluble HVEM might have diagnostic and prognostic value in certain immunological disorders.