Suppression of Proinflammatory Cytokines and Mediators in LPS-Induced RAW 264.7 Macrophages by Stem Extract of Alternanthera sessilis via the Inhibition of the NF-κB Pathway.

Alternanthera sessilis, an edible succulent herb, has been widely used as herbal drug in many regions around the globe. Inflammation is a natural process of the innate immune system, accompanied with the increase in the level of proinflammatory mediators, for example, nitric oxide (NO) and prostaglandin (PGE2); cytokines such as interleukin 6 (IL-6), interleukin 1ß (IL-1ß), and tumor necrosis factor alpha (TNFα); and enzymes including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) via the activation and nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) subunit p65 due to the phosphorylation of inhibitory protein, IκBα. Inflammation over a short period of time is essential for its therapeutic effect. However, prolonged inflammation can be detrimental as it is related to many chronic diseases such as delayed wound healing, cardiovascular disease, arthritis, and autoimmune disorders. Therefore, ways to curb chronic inflammation have been extensively investigated. In line with that, in this present study, we attempted to study the suppression activity of the proinflammatory cytokines and mediators as a characteristic of anti-inflammatory action, by using stem extract of A. sessilis in the lipopolysaccharide- (LPS-) stimulated RAW 264.7 macrophage cell line. The results showed that the extract has significantly inhibited the production of the proinflammatory mediators including NO and PGE2; cytokines comprising IL-6, IL-1ß, and TNFα; and enzymes covering the iNOS and COX-2 by preventing the IκBα from being degraded, to inhibit the nuclear translocation of NF-κB subunit p65 in order to hinder the inflammatory pathway activation. These results indicated that the stem extract of A. sessilis could be an effective candidate for ameliorating inflammatory-associated complications.

Amaranthaceae pollens: review of an emerging allergy in the mediterranean area.

The Amaranthaceae family is composed of about 180 genera and 2500 species. These common weeds have become increasingly relevant as triggers of allergy in the last few years, as they are able to rapidly colonize salty and arid soils in extensive desert areas. The genera Chenopodium, Salsola, and Amaranthus are the major sources of pollinosis from the Amaranthaceae family in southern Europe, western United States, and semidesert areas of Saudi Arabia, Kuwait, and Iran. In Spain, Salsola kali is one of the most relevant causes of pollinosis, together with olive and grasses. To date, 9Amaranthaceae pollen allergens from Chenopodium album, Salsola kali, and Amaranthus retroflexus have been described and are listed in the International Union of Immunological Societies allergen nomenclature database.The major allergens ofAmaranthaceae pollen belong to the pectin methylesterase, Ole e 1-like, and profilin panallergen families, whereas the minor allergens belong to the cobalamin- independent methionine synthase and polcalcin panallergen families. These relevant allergens have been characterized physicochemically, and immunologically at different levels. Recombinant forms, allergenic fusion recombinant proteins, and hypoallergenic derivatives of these allergens have been expressed in bacteria and yeast and compared with their natural proteins from pollen. In this review, we provide an extensive overview ofAmaranthaceae pollen allergens, focusing on their physicochemical, and immunological properties and on their clinical significance in allergic patients. We also review studies where these recombinant allergens and their hypoallergenic derivatives have been used in clinical diagnosis and their potential use in personalized therapy.

Assessing degree of flowering implicates multiple Chenopodiaceae/Amaranthaceae species in allergy.

BACKGROUND: IgE-mediated sensitization to the Chenopodiaceae/Amaranthaceae families is a cause of allergic symptoms in arid areas. Salsola kali and Chenopodium album are considered the main species responsible; however, there is a discrepancy between the pollination period of these two plants and clinical symptoms. The objectives of this study were to identify new Chenopodiaceae/Amaranthaceae members with sensitization capacity and to correlate symptoms, pollen counts and degree of flowering of different species. METHODS: A total of 37 individuals monosensitized to S. kali and C. album were included in the study. All patients recorded daily symptom scores between May and October 2007. Extracts from Chenopodium (album, vulvaria and murale), Salsola (kali, vermiculata, and oppositifolia), Bassia scoparia, Atriplex (patula and halimus) and Amaranthus (deflexus and muricatus) were manufactured and used in skin prick tests (SPTs). Protein content and IgE binding were assessed for each extract. Pollen counts and degree of flowering (based on the Orshan specific semiquantitative method) were assessed weekly. RESULTS: Symptom scores demonstrated a positive correlation with pollen counts even outside the pollination period of S. kali. Positive SPTs were obtained with all 11 species tested, which showed common proteins with IgE-binding capacity. Different species flowered at different times during the pollen season. CONCLUSION: Different taxonomically related species of Chenopodiaceae/Amaranthaceae can induce allergic sensitization and should be considered for use in diagnosis and treatment. Degree of flowering is a complementary method for assessing pollination that could be used for botanical families with indistinguishable pollen grains.

Immunochemical characterization of Amaranthus retroflexus pollen extract: extensive cross-reactive allergenic components among the four species of Amaranthaceae/Chenopodiaceae.

The importance of Amaranthus retroflexus pollen in causing respiratory allergy has been well ascertained in many countries including Iran with a high positive rate (69%) among Iranian allergic patients. The aim of the present study is to identify the allergenic properties of A. retroflexus pollen. Sixteen patients with allergy to A. retroflexus pollen were selected for the study. The antigenic and allergenic profiles of the A. retroflexus pollen extract as well as pollen extracts from other species of the Amaranthaceae/Chenopodiaceae family, including Chenopodium album, Kochia scoparia, and Salsola kali, were evaluated by ELISA, immunoblotting, and immunoblot inhibition assays. The resolved protein fractions on SDS-PAGE ranged from 10-85 kDa. Several allergenic components (MW 85, 45, 39, 18, 15, and 10 kDa) of the A. retroflexus pollen extract were recognized by using patients' sera by specific antibody of IgE class using ELISA and immunoblot assays. The IgE reactivity of the A. retroflexus pollen extract was partially inhibited by all three pollen extracts tested. The inhibition by the S. kali pollen extract was more than those by other pollen extracts. Moreover, the wheal diameters by the A. retroflexus pollen extract were highly correlated with those by C. album, K. scoparia and S. kali pollen extracts. In conclusion, three proteins with apparent MWs of 39, 45, and 66 kDa are suggested as the common allergenic components among the four pollens from the Amaranthaceae/Chenopodiaceae family. It appears that there are some common (similar) epitopes among the four common allergenic pollens.

Correlation between Chenopodiacea/Amaranthacea pollen counts and allergic symptoms in Salsola kali monosensitized patients.

UNLABELLED: We performed a prospective observational study to establish a relationship between pollen counts of Chenopodiacea/Amaranthacea and clinical symptoms of rhinoconjunctivitis and asthma in a group of monosensitised patients. MATERIAL AND METHODS: A total of 60 patients (19 with asthma) were included in the study. All patients collected daily symptom scores during the summer months of 1999, 2000 and 2001. The questionnaire included ocular, nasal and pulmonary symptoms. Pollen counts were expressed as pollen grains/m3. Symptom scores and pollen counts were correlated using correlation coefficients and Log transformed variables. RESULTS: In the 3 seasons studied we identified a peak of pollen and clinical symptoms in the second half of August and first half of September. In 1999, there was a significant positive correlation between total symptoms and daily pollen grains/m3 (p<0.005, r = 0.347). This correlation was not significant for the summers of 2000 and 2001. After further analysis, and by displacing one of both variables between 11 to 17 days, the correlation coefficients for total symptoms, improved for 1999 (r = 0. 744; p < 0.0001) and became significant for 2000 (r = 0. 521; p < 0.0001) and 2001 (r = 0.635; p < 0.0001). CONCLUSION: We identified a significant time lag between pollen counts and symptom scores in S. kali monosensitized patients.

A monoclonal antibody to feruloylated-(1-->4)-beta-D-galactan.

We report the isolation and characterization of a monoclonal antibody, designated LM9, against feruloylated-(1-->4)-beta-D-galactan. This epitope is a structural feature of cell wall pectic polysaccharides of plants belonging to the family Amaranthaceae (including the Chenopodiaceae). Immuno-assays and immunofluorescence microscopy indicated that LM9 binding is specific to samples and cell walls obtained from species belonging to this family. In a series of competitive-inhibition enzyme-linked immunosorbent assays with potential oligosaccharide haptens, the most effective inhibitor was O-[6-O-(trans-feruloyl)-beta-D-galactopyranosyl]-(1-->4)-D-galactopyranose (Gal2F). LM9 is therefore a useful antibody probe for the analysis of phenolic substitution of cell wall pectic polymers and of cell wall structure in the Amaranthaceae including sugar beet (Beta vulgaris L.) and spinach (Spinacia oleracea L.).

The seasonal variation in allergic rhinitis and its correlation with outdoor allergens in Kuwait.

BACKGROUND: Allergic rhinitis (AR) is the most common allergic problem in Kuwait. Most of the patients who have either AR or asthma are referred to the Al-Rashed Allergy Center. OBJECTIVE: To determine if there is a seasonal variation in AR in Kuwait and to correlate it with the daily pollen count. METHODS: Information about the new patients referred to the center over a 5-year study period (1996-2000) was extracted from the center's records. The daily pollen count in Kuwait city was obtained from the Air Biology Laboratory. RESULTS: There was a significant seasonal variation with a bimodal increase in the number of patients with AR referred to the center. The main peak in the number of patients occurred in September-October, and there was a smaller peak in April-May. The mean number +/- SD of new patients per month over the 5-year period varied from 87 +/- 32 for December to 367 +/- 104 for September. Similarly, the average daily pollen count varied from 3.7 +/- 1.0 pollens per mm3 in January to 124 +/- 92 in October. There was high correlation between the number of new AR patients and the average total pollen count (Pearson correlation, r=0.77, p<0.001), as well as with Chenopodiaceae and Amaranthaceae (weed) pollens (r=0.75, p<0.001), while there was no correlation between the number of new patients and either tree or grass pollens. CONCLUSION: Seasonal AR occurs during two periods in Kuwait, i.e. September-October and April-May, with September-October being the main season. The rise in AR during late summer in Kuwait is mainly associated with the pollination of Chenopodiaceae species.

Immunological characteristics of PEP carboxylase from leaves of C3-, C4- and C3-C4 intermediate species of Alternanthera--comparison with selected C3- and C4- plants.

Immunological cross-reactivity of phosphoenolpyruvate carboxylase (PEPC) in leaf extracts of C3-, C4- and C3-C4 intermediate species of Alternanthera (along with a few other C3- and C4- plants) was studied using anti-PEPC antibodies raised against PEPC of Amaranthus hypochondriacus (belonging to the same family as that of Alternanthera, namely Amaranthaceae). Antibodies were also raised in rabbits against the purified PEPC from Zea mays (C4- monocot-Poaceae) as well as Alternanthera pungens (C4- dicot-Amaranthaceae). Monospecificity of PEPC-antiserum was confirmed by immunoprecipitation. Amount of PEPC protein in leaf extracts of A. hypochondriacus could be quantified by single radial immunodiffusion. Cros- reactivity of PEPC in leaf extracts from selected C3-, C4-, and C3-C4 intermediate species (including those of Alternanthera) was examined using Ouchterlony double diffusion and Western blots. Anti-PEPC antiserum raised against A. hypochondriacus enzyme showed high cross-reactivity with PEPC in leaf extracts of A. hypochondriacus or Amaranthus viridis or Alternanthera pungens (all C4 dicots), but limited cross-reactivity with that of Zea mays, Sorghum or Pennisetum (all C4 monocots). Interestingly, PEPC in leaf extracts of Alternanthera tenella, A. ficoides, Parthenium hysterophorus (C3-C4 intermediates) exhibited stronger cross-reactivity (with anti-serum raised against PEPC from Amaranthus hypochondriacus) than that of Pisum sativum, Commelina benghalensis, Altenanthera sessilis (C3 plants). Further studies on cross-reactivities of PEPC in leaf extracts of these plants with anti-PEPC antisera raised against PEPC from leaves of Zea mays or Alternanthera pungens confirmed two points--(i) PEPC of C3-C4 intermediate is distinct from C3 species and intermediate between those of C3- and C4-species; and (ii) PEPC of C4-dicots was closer to that of C3-species or C3-C4 intermediates (dicots) than to that of C4-monocots.