Immunodetection of Pectic Epitopes, Arabinogalactan Proteins, and Extensins in Mucilage Cells from the Ovules of Pilosella officinarum Vaill. and Taraxacum officinale Agg. (Asteraceae).

The main aim of this study was to compare the cytological difference between ovular mucilage cells in two Asteraceae species-Pilosella officinarum and Taraxacum officinale-in order to determine whether pectic epitopes, arabinogalactan proteins, or extensins are present. The immunocytochemical technique was used. Both the Taracacum and Pilosella genera have been used recently as models for understanding the mechanisms of apomixis. Knowledge of the presence of signal molecules (pectic epitopes, arabinogalactan proteins, and extensins) can help better understand the developmental processes in these plants during seed growth. The results showed that in Pilosella officinarum, there was an accumulation of pectins in the mucilage, including both weakly and highly esterified pectins, which was in contrast to the mucilage of Taraxacum officinale, which had low amounts of these pectins. However, Taraxacum protoplasts of mucilage cells were rich in weakly methyl-esterified pectins. While the mucilage contained arabinogalactan proteins in both of the studied species, the types of arabinogalactan proteins were different. In both of the studied species, extensins were recorded in the transmitting tissues. Arabinogalactan proteins as well as weakly and highly esterified pectins and extensins occurred in close proximity to calcium oxalate crystals in both Taraxacum and Pilosella cells.

Asteraceae species as potential environmental factors of allergy.

The statistics from Europe and the USA have proven a high risk for skin diseases associated with plant contact. Therefore, plant-induced dermatitis is of increasing attention in dermatology. The focus of this paper was to present the current knowledge on aspects of contact allergy related to Asteraceae (Compositae) species. The Asteraceae family is one of the largest in the world with members across all continents. The PubMed/Medline databases have been searched. The Asteraceae representatives consist of diverse secondary metabolites, which exhibit various advantageous effects in humans. In particular, sesquiterpene lactones (SLs) may cause sensitization resulting in skin irritation and inflammation. In this study, we tried to reveal the allergenic potential of several Asteraceae species. The Asteraceae-related allergy symptoms involve eczema, hay fever, asthma, or even anaphylaxis. Furthermore, the evidence of severe cross-reactivity with food and pollen allergens (PFS) in patients sensitive to Asteraceae allergens have been announced. Further identification and characterization of secondary metabolites and possible allergens in Asteraceae are necessary for the better understanding of Asteraceae-related immune response. The Asteraceae allergy screening panel (the SL mix and the Compositae mix of five plant species) is a promising tool to improve allergy diagnostics and therapy.

Anti-inflammatory and antioxidant potential of Guaianolide isolated from Cyathocline purpurea: Role of COX-2 inhibition.

BACKGROUND: Inflammation activated by oxidative stress can cause various diseases, such as asthma, rheumatoid arthritis, cancer, diabetes, etc. Plant constituents with sesquiterpene lactones possess antioxidant and anti-inflammatory properties. AIM: To determine the antioxidant and anti-inflammatory potential of isolated phytoconstituent from Cyathocline purpurea Buch-Ham ex D (CP). Don in laboratory animals. Furthermore, to understand the interactions involved in the binding of this compound to cyclooxygenase-2 (COX-2) via computational docking. METHODS: Phytoconstituent was isolated, purified and well characterized (using IR, NMR, and MS) from ethyl acetate fraction of CP methanolic extract. It was then evaluated for its in-vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) and hydroxyl (OH) radical assays as well as in-vivo anti-inflammatory potential against carrageenan-induced paw edema model in rats. The molecular docking study was performed against the crystal structure of COX-2 to evaluate the binding potential of phytoconstituent towards this enzyme. RESULTS: The isolated compound 6α-hydroxy-4 [14], 10 [15]-guainadien-8α, 12-olide (HGN) showed significant (p<0.001) antioxidant activity with IC50 values of 76µg/mL. Administration of HGN (10 and 20mg/kg) significantly (p<0.001) reduced the increased paw volume after subplantar administration of carrageenan. It also exhibits good binding affinity towards with COX-2 with a docking score of -8.98 and Glide binding energy of -36.488kcal/mol shedding light on the potential mechanism of anti-inflammatory action. CONCLUSIONS: The presence of hydroxyl group in HGN provides a credential to its in-vivo anti-inflammatory and in-vitro antioxidant activities. Furthermore, the good binding affinity of HGN for the active site of COX-2 may open novel vistas in therapeutic option with natural antioxidants like Cyathocline purpurea to treat various inflammatory disorders.

Performance of a commercially available plant allergen series in the assessment of suspected occupational contact dermatitis to plants in north Indian patients.

BACKGROUND: Parthenium hysterophorus is the leading cause of phytogenic allergic contact dermatitis in India. The Indian Standard Series currently supplied by Systopic Laboratories Ltd and manufactured by Chemotechnique Diagnostics ® contains parthenolide as the only allergen representing plant allergens. AIM: The study was conducted to assess the performance of the Chemotechnique plant series (PL-1000), consisting of 14 allergens, in patients with clinically suspected occupational contact dermatitis to plant allergens. METHODS: Ninety patients were patch tested with the Chemotechnique plant series from 2011 to 2013. Demographic details, clinical diagnosis and patch test results were recorded in the contact dermatitis clinic proforma. RESULTS: Of 90 patients, 24 (26.7%) showed positive reactions to one or more allergens in the plant series. Positive patch tests were elicited most commonly by sesquiterpene lactone mix in 19 (78.6%) patients, followed by parthenolide in 14 (57.1%), Achillea millefolium in 10 (42.9%) and others in decreasing order. CONCLUSION: The plant allergen series prepared by Chemotechnique Diagnostics is possibly not optimal for diagnosing suspected allergic contact dermatitis to plants in north Indians. Sesquiterpene lactone mix should replace parthenolide as the plant allergen in the Indian Standard Series until relevant native plant extracts are commercially available for patch testing.

The effect of 4α,5α-epoxy-10α,14-dihydro-inuviscolide, a novel immunosuppressant isolated from Carpesium abrotanoides, on the cytokine profile in vitro and in vivo.

The plant Carpesium abrotanoides (CA) is used in Asian herbal medicines as an insecticide and to treat bruises. However, the effect of single compounds from CA blooms and the mechanism of its immunosuppressive effect remain poorly understood. The aim of this study was to investigate the mechanism of the immunosuppressive effect in the three kinds of immune cells, and the immunosuppressive effect of CA bloom extract (CAE) in acute inflammation models (LPS and ConA-induced inflammation). Interleukin-6, IL-4, IL-13, IFNγ, and IL-10-but not TNFα-were significantly reduced in a dose-dependent manner by 4α,5α-epoxy-10α,14-dihydro-inuviscolide (INV). Furthermore, INV inhibited NF-κB transcriptional activation and IL-10 promoter activity in the same manner as for Bay11. Meanwhile, treatment with dexamethasone reduced the levels of IFNγ, but not IL-10, and resulted in no change in NF-κB transcriptional activation or the IL-10 promoter. INV did not affect PMA-induced IκB kinase complex phosphorylation, IκB degradation, or MAPK and the nuclear translocation of p65, as with DEX. The in vivo, CAE has an immunosuppressive effect on the LPS-induced inflammation response model by inhibiting the plasma level of IFNγ and IL-6 levels. CAE treatment also tends to attenuate the plasma level of IFNγ, IL-4, and IL-6 in ConA-induced inflammation. These findings indicate that INV causes the reduction of the cytokine profile by blocking the NF-κB transcription factor activation and the molecular mechanism by which INV operates could provide new insights into the unique mechanisms responsible for NF-κB inhibition, in contrast to established immunosuppressants, as a therapeutic agent for immunopathological treatment.

Genetic endowment of proinflammatory cytokine tumor necrosis factor-α (-) 308 G > A polymorphism to Parthenium hysterophorus-induced airborne contact dermatitis in an Indian cohort.

BACKGROUND: Parthenium dermatitis is a common airborne allergic health problem that induces a cell-mediated hypersensitivity immune response involving activated T lymphocytes, which culminates in injury to the skin. The disease is manifested as itchy erythematous papules and plaques and primarily affects the exposed areas and flexures. This study aimed to identify the role of tumor necrosis factor (TNF)-α (-) 308 G>A polymorphism in the pathogenesis of parthenium dermatitis. MATERIALS AND METHODS: A total of 120 subjects, including 60 patients exclusively diagnosed for parthenium dermatitis and 60 healthy individuals, were included in the study. The genotyping of the TNF-α (-) 308 G>A region was carried out by the amplification refractory mutational system. RESULTS: In the present study, we demonstrated that polymorphism of the TNF-α (-) 308 position (A and/or G) was not statistically significant, and there was no difference in the distribution of any alleles of this locus in cases and controls. CONCLUSION: The present study suggests that there is a lack of association of potent proinflammatory cytokine TNF-α (-) 308 G>A polymorphism in parthenium dermatitis in the Indian cohort. It interprets genetically endowed transcriptional capacity due to this particular single nucleotide polymorphism but does not support the prevalence of high serum levels of TNF-α in parthenium-induced skin allergic inflammation.

Plantas medicinales utilizadas en las comunidades de Itá Azul y San Gervasio (Paraguay) / Plantas medicinais utilizadas nas comunidades de Ita Azul e San Gervasio (Paraguai) / Medicinal plants used in the communities of Ita Azul and San Gervasio (Paraguay)

El presente trabajo se desarrolló dentro del marco del Proyecto: «Conservación, fortalecimiento y uso sostenible de la flora de Itá Azul y San Gervasio, Colonia Independencia, Reserva de Recursos Manejados Yvyturuzú. Paraguay», tuvo por objetivo determinar las plantas medicinales empleadas por las comunidades de Itá Azul y San Gervasio, (Colonia Independencia, Departamento de Guairá), Paraguay. Para ello se realizaron encuestas en las mencionadas comunidades, siendo encuestadas en total 81 personas, quienes reportaron un total de 68 especies de uso medicinal distribuidas en 35 familias, siendo las familias con mayor número de especies las Asteráceas (19%), Rutáceas (9%) y Fabáceas, Lamiáceas y Verbenáceas (6% cada una). El 53 % de las especies reportadas son nativas. Se prefiere el uso de las plantas en estado fresco (36%), siendo la parte de la planta más empleada la hoja (32%) y la forma de uso mayormente mencionada el tereré (infusión de las plantas conjuntamente con hoja de mate, 43%). Las afecciones reportadas se agruparon en 11 categorías, de las cuales los trastornos digestivos seguidos por las afecciones respiratorias fueron las más reportadas en el uso de plantas medicinales. Para 11 especies no se ha reportado el uso. El conocimiento etnofarmacobotánico se transmite oralmente de una generación a otra, principalmente a través de las mujeres. La investigación científica puede contribuir eficazmente a preservar este patrimonio cultural (AU)

O presente trabalho foi desenvolvido no âmbito do projeto "Conservación, Fortalecimiento y Uso Sostenible de la flora de Itá Azul y San Gervasio, Colonia Independencia, Reserva de Recursos Manejados Yvyturuzú. Paraguay", com o objetivo de determinar as plantas medicinais utilizadas pelas comunidades de Ita Azul e San Gervasio, Colonia Independencia, Departamento de Guairá, no Paraguai. Para isso foram realizados inquéritos nas comunidades acima mencionadas, tendo sido inquiridas um total de 81 pessoas, que reportaram um total de 68 espécies de uso medicinal, distribuídas em 35 famílias, sendo as famílias com maior número de espécies as Asteraceae (19%), Rutaceae (9 %), seguindo-se Fabaceae, Lamiaceae e Verbenaceae (com 6% cada uma). Cerca de 53% das espécies reportadas são nativas. Utilizamse preferencialmente plantas frescas (36%), sendo a parte da planta mais utilizada a folha (32%) e a forma de uso principalmente mencionada o tereré (43%). As afeções reportadas foram agrupadas em 11 categorias, das quais os distúrbios digestivos foram os mais relatados sobre o uso de plantas medicinais e em segundo lugar as afeções respiratórias. Para 11 espécies não foi reportado o seu uso. O conhecimento etnofarmacobotánico é transmitido oralmente de uma geração para outra, principalmente através das mulheres. A investigação científica pode contribuir de forma eficaz para preservar este património cultural

The present work was developed within the framework of the project «Conservation, strengthening and sustainable use of the flora of Ita Azul and San Gervasio, Colonia In dependencia, Managed Resources Reserve Yvyturuzú. Paraguay», aimed to determine the medicinal plants used by the communities of Ita Azul and San Gervasio, Colonia Independencia, Guairá Department, Paraguay. Surveys were conducted in the aforementioned communities to 81 people, who reported the use of 68 species of medicinal plants belonging to 35 families, mainly to Asteraceae (19%), Rutaceae (9 %), as well as Fabaceae, Lamiaceae and Verbenaceae (6% each). 53% of the species reported were native. The use of fresh plants (36%) was preferred, being the leaf the part of the plant most frequently used (32%). The plants were mainly prepared as infusion, mixed with mate leaf, in a preparation which is known as «tereré» (43%). The conditions for which the medicinal plants were used were grouped in 11 categories, being digestive disorders followed by respiratory conditions the most reported. For 11 species, no use was reported. The ethnofarmacological and ethnobotanical knowledge is transmitted orally from one generation to another, mainly by women. Scientific research can contribute efficiently to preserve this cultural heritage (AU)

Anti-inflammatory and antioxidant activities of the methanol extract of Gynura segetum leaf.

Gynura segetum, family Compositae, is a cultivated species and can be found growing in the tropical regions of Indonesia and Malaysia. The plant is known for its use for the treatment of cancer, inflammation, diabetes, hypertension and skin afflictions. In the current study, in vivo anti-inflammatory effect of the methanol extract G. segetum leaf and its antioxidant effect in vitro have been investigated for the first time. The in vitro antioxidant activities of the methanol extract were measured using common methods including total phenolic content; total flavonoid content; scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ß-carotene bleaching assays. The in vivo anti-inflammatory activities were tested using the cotton pellet implanted animal model. The measurement of pro-inflammatory cytokine (TNF-α and IL-1) levels in the blood samples of the rats was carried out by using ELISA kits. The inhibitory activity on cyclooxygenase (COX) enzyme of methanol extract was also evaluated. The methanol extract exhibited good antioxidant activity which is associated with their total phenolic and flavonoid contents. Methanol extract strongly inhibited the granuloma tissue formation in rats and the anti-inflammatory potential was mediated through the inhibition of pro-inflammatory cytokines and COX-2 enzyme activities. Taken together, the present study suggests that G. segetum's leaf is a natural source of antioxidants and has potential therapeutic benefits against chronic inflammation.

Allergens of weed pollen: an overview on recombinant and natural molecules.

Weeds represent a botanically unrelated group of plants that usually lack commercial or aesthetical value. Pollen of allergenic weeds are able to trigger type I reactions in allergic patients and can be found in the plant families of Asteraceae, Amaranthaceae, Plantaginaceae, Urticaceae, and Euphorbiaceae. To date, 34 weed pollen allergens are listed in the IUIS allergen nomenclature database, which were physicochemically and immunologically characterized to varying degrees. Relevant allergens of weeds belong to the pectate lyase family, defensin-like family, Ole e 1-like family, non-specific lipid transfer protein 1 family and the pan-allergens profilin and polcalcins. This review provides an overview on weed pollen allergens primarily focusing on the molecular level. In particular, the characteristics and properties of purified recombinant allergens and hypoallergenic derivatives are described and their potential use in diagnosis and therapy of weed pollen allergy is discussed.

Natural rubber biosynthesis in plants: rubber transferase.

Rubber biosynthesis in plants is a fascinating biochemical system, which evolved at the dawn of the dicotyledoneae and is present in at least four of the dictolydonous superorders. Rubber biosynthesis is catalyzed by a membrane complex in a monolayer membrane envelope, requires two distinct substrates and a divalent cation cofactor, and produces a high-molecular-weight isoprenoid polymer. A solid understanding of this system underpins valuable papers in the literature. However, the published literature is rife with unreliable reports in which the investigators have fallen into traps created by the current incomplete understanding of the biochemistry of rubber synthesis. In this chapter, we attempt to guide both new and more established researchers around these pitfalls.

Parthenium dermatitis in India: past, present and future.

Parthenium dermatitis is an immuno-inflammatory disease caused by Parthenium hysterophorus and is the commonest cause of plant dermatitis in India. It is caused by airborne dry and friable plant particles including trichomes, and the most important allergens responsible for allergic contact dermatitis are sesquiterpene lactones. The combined type IV and type I hypersensitivity to parthenium has been recently postulated. In sensitized individuals, it can cause a spectrum of clinical patterns, such as classical airborne pattern, chronic actinic dermatitis-like presentation, mixed pattern dermatitis, exfoliative dermatitis, widespread dermatitis, and other rare patterns. There is definite trend towards change from airborne pattern to chronic actinic pattern in natural history of parthenium dermatitis. Contact sensitivity to parthenium is everlasting, and hence the disease runs a chronic course with exacerbation during summers. Patch testing with acetone or aqueous plant extract is the simplest way of confirming parthenium contact allergy. Management includes avoiding contact with allergen, managing dermatitis with topical corticosteroids/tacrolimus, and other immunosupressives like azathioprine. In future, we expect parthenium dermatitis to become less prevalent due to rapid urbanization and possible development of new biological methods to eradicate the parthenium. Genetic factors associated with susceptibility to parthenium dermatitis need to be studied.

Immunotoxic sesquiterpene lactone from Carpesium rosulatum Miq.

The whole plants of Carpesium rosulatum were chloroform extracted and the isolated sesquiterpene lactones and immunotoxicity effects were studied. The structures and stereochemistry of these compounds were established on the basis of analysis of spectra including mp, [α](D)(25), IR, UV, EI-MS, MS, (1)H-NMR, (13)C-NMR and some chemical transformations as follows: 1 (4ß,10α-dihydroxy-guaia-8α,12-olide), 2 (4ß,10α-dihydroxy-1(2),11 (13)-guaiadien -8α,12-olide), 3 (3ß,8ß-dihydroxy-1α,5α-guaian-10(14)-ene-6α,12-olide). 4 (2ß,5-epoxy-5,10-dihydroxy-6α,9ß-diangeloyloxy-germacran-8α,12-olide) The chloroform extracted had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L with an LC(50) value of 13.11 ppm and an LC(90) value of 20.33 ppm. The results could be useful in search for newer, safer, and more effective natural immunotoxicity agents against A. aegypti.

Study of pro- and anti-inflammatory cytokine profile in the patients with parthenium dermatitis.

BACKGROUND: Parthenium dermatitis is a common airborne allergic contact dermatitis induced by exposures to the weed Parthenium hysterophorus. The disease manifests as itchy erythematous papules, papulovesicular and plaque lesions on exposed areas of the body. OBJECTIVES: The aim of this study was to show the alterations in pro/anti-inflammatory cytokines in parthenium dermatitis. METHODS: The study included 50 patients with parthenium dermatitis confirmed by patch testing using aqueous extracts of P. hysterophorus and 50 age-matched healthy controls. The levels of pro-inflammatory [tumour necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-17] and anti-inflammatory (IL-4 and IL-10) cytokines were estimated by commercially available high sensitivity enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: All the dermatitis patients showed significantly (P < 0.001) elevated levels of TNF-α, IL-6, IL-8, and IL-17 levels as compared to healthy controls. In contrast, the anti-inflammatory cytokine IL-4 showed an insignificant decrease (P < 0.217) and a decrease in level of IL-10 was statistically significant (0.001) compared with controls. CONCLUSIONS: The present study suggests the involvement of pro-inflammatory cytokines in the pathogenesis of parthenium dermatitis. A decrease in levels of anti-inflammatory cytokines was demonstrated, which could not downregulate pro-inflammatory cytokines in parthenium dermatitis.

Anti-allergic effects of fermented Ixeris sonchifolia and its constituent in mice.

To evaluate the antiallergic effect of fermented Ixeris sonchifolia (IS, family Compositae), we prepared IS Kimchi, isolated Lactic acid bacteria (LAB) from it, fermented IS with these LAB, and investigated their antiallergic effects. IS Kimchi more potently inhibited the passive cutaneous anaphylaxis (PCA) reaction induced by an IgE-antigen complex as well as the scratching behavior induced by compound 48/80 or histamine than IS. When IS was fermented with LAB isolated from IS Kimchi, its antiallergic effects was also increased. Of LAB used for fermentation, Lactobacillus brevis more potently increased the antiallergic effects. Its main constituents, chlorogenic acid and luteolin potently inhibited PCA reaction induced by IgE-antigen complex as well as pruritus induced by compound 48/80 or histamine. These constituents inhibited the expression of proinflammatory and allergic cytokines, TNF-alpha and IL-4, and transcription factor, NF-kappaB, activation induced by IgE-antigen complex in RBL-2H3 cells, as well as the degranulation of RBL-2H3 cells induced by an IgE-antigen complex. Luteolin more potently inhibited these allergic reactions than chlorogenic acid. These findings suggest that antiallergic effect of IS can be increased by LAB fermentation and fermented IS might improve allergic reactions, such as pruritus, anaphylaxis, and inflammation.

New aspects in allergic contact dermatitis.

PURPOSE OF REVIEW: To give selected new information on contact allergy and allergic contact dermatitis with focus on diagnostic procedures and pitfalls. RECENT FINDINGS: Recent studies dealing with common contact allergens have improved our understanding of the relationship between positive patch tests and the clinical interpretation and consequences for the patient. SUMMARY: Nickel allergy is still the most common contact allergy in Europe in spite of full implementation of the EU Nickel Directive in 2001. Contact allergens in cosmetics and topical drugs are another common cause of allergic contact dermatitis. The main culprits include fragrance chemicals, preservatives, and hair dyes. We are all more or less exposed to cosmetics and topical drugs on a daily basis. The labelling requirements given in the Cosmetics Directive is of great help in tracing the causative allergenic ingredients. Most of the components present in cosmetic products are also occurring in household and industrial products, often under other trade names. Patients with multiple contact allergies constitute a special problem because their quality of life is severely affected by the multitude of eliciting products in the environment. We still lack a good understanding of why these patients become so easily sensitized.

Allergenicity and cross-reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach.

BACKGROUND: The genus Senecio is the largest genus of the family Asteraceae (Compositae). The allergenicity of Senecio has not been assessed previously. OBJECTIVE: The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance. METHODS: Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one-dimensional electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross-reactivity with other pollen. RESULTS: Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human-allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two-dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross-reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies. CONCLUSION: S. jacobea constitute a newly discovered allergenic source. It shows cross-reactivity with other members of the Asteraceae plant family as well as with P. judaica.