Aqueous Phase Extracts Research Articles

Non-targeted chemical analysis of consumer botanical products labeled as blue cohosh (Caulophyllum thalictroides), goldenseal (Hydrastis canadensis), or yohimbe bark (Pausinystalia yohimbe) by NMR and MS.

Consumers have unprecedented access to botanical dietary supplements through online retailers, making it difficult to ensure product quality and authenticity. Therefore, methods to survey and compare chemical compositions across botanical products are needed. Nuclear magnetic resonance (NMR) spectroscopy and non-targeted mass spectrometry (MS) were used to chemically analyze commercial products labeled as containing one of three botanicals: blue cohosh, goldenseal, and yohimbe bark. Aqueous and organic phase extracts were prepared and analyzed in tandem with NMR followed by MS. We processed the non-targeted data using multivariate statistics to analyze the compositional similarity across extracts. In each case, there were several product outliers that were identified using principal component analysis (PCA). Evaluation of select known constituents proved useful to contextualize PCA subgroups, which in some cases supported or refuted product authenticity. The NMR and MS data reached similar conclusions independently but were also complementary.

Novel isolate of Cladosporium subuliforme and its potential to control Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae)

BackgroundCitrus fruits are economically and nutritionally important but have been severely affected by Huanglongbing disease (HLB), its natural spread is mainly by the Asian Citrus Psyllid,Diaphorina citri Kuwayama (Hemiptera: Liviidae). Chemicals are often used to control this pest, but this is not sustainable. Meanwhile, there are few environmentally friendly bioinsecticides to control D. citri in China.ResultsIn this study, an entomofungal pathogen wz-1 was isolated from a D. citri carcass in the field, which resulted in a cumulative mortality rate of 75.27% in adult D. citri 7 days after inoculation of the spore suspension. It was identified as Cladosporium subuliforme based on morphological analysis as well as sequence analysis of several molecular markers (Internal Transcribed Spacers, Translation Elongation Factor 1-α and Actin). Remarkably, the lethality rate of adult D. citri reached 53.13%, 48 h after treatment with the aqueous phase extracts of wz-1. Hydroxyquinoline and phytosphingosine in the extracts were identified as potentially active metabolites using LC–MS.ConclusionsThe entomopathogenicity and bioinsecticidal potential of C. subuliforme were previously unknown. Obtained results showed that both spores and extracts of wz-1 can effectively kill adult D. citri, providing an available fungal resource and a theoretical basis for biocontrol of the HLB insect vector D. citri.

Assessment of weed root extracts for allelopathic activity against Orobanche and Phelipanche species

Broomrapes (Orobanche and Phelipanche species) are holoparasitic weeds that infect roots of crop hosts from Asteraceae, Brassicaceae, Apiaceae, Fabaceae, and Solanaceae. The parasitic weeds are difficult to control selectively without crop damage once attached to their roots. Identification of natural compounds with herbicidal activity against pre-attached broomrape stages can provide control alternatives. With the aim to identify plant species with efficacy for broomrape control, organic and residual aqueous phase extracts from roots of seventeen weed species common in south Spanish broomrape-infested fields were assessed as potential inducers of suicidal broomrape germination and as inhibitors of broomrape radicle growth. Assessments were carried out in vitro using seeds and seedlings of four noxious broomrape species, Orobanche crenata Forsk., Orobanche cumana Wallr., Orobanche minor Sm. and Phelipanche ramosa (L.) Pomel. While root extracts from all the weed species did not induce suicidal germination on O. crenata seeds, most of the extracts induced germination of P. ramosa except for those obtained from Amaranthus albus L., Amaranthus retroflexus L. and Convolvulus arvensis L. Moderate levels of germination activity were induced in O. cumana and O. minor seeds by some of the root extracts tested, with strongest induction obtained from Heliotropium europaeum L. on O. cumana seeds, and from Silybum marianum (L.) Gaertn. on O. minor seeds. For root extract inhibition of broomrape radicles, the extract from C. arvensis roots strongly inhibited radicles of all the broomrape species. While extracts from the other weed species induced low or negligible inhibition of O. cumana and O. crenata radicle growth, many inhibited P. ramosa and O. minor radicles. Exceptions were root extracts from Datura stramonium L., Heliotropium europaeum L., Malva sylvestris L., Solanum nigrum L. and Urtica dioica L., which did not inhibit P. ramosa radicles, and those from A. retroflexus, Datura stramonium L., Malva sylvestris L., Portulaca oleracea L. and S. nigrum, which did not inhibit O. minor radicles. Among the active organic extracts assessed, those showing promising chemical profiles were selected for future studies to characterize natural compounds with potential herbicidal activity on early stages of broomrape growth.

Abstract 2353: Metabolic reprogramming by SLC1A5 downregulation in pancreatic cancer cells

Abstract Introduction: Pancreatic cancers exhibit limited response to chemo- and radiation therapy. Identifying novel targets for pancreatic cancer treatment is important for new treatment strategies. Because pancreatic cancer cells are ‘glutamine avid' [1], targeting glutamine metabolic pathways can provide novel options for treatment. The glutamine transporter, SLC1A5, is being actively investigated as a pharmacological target in cancer [2]. Here we have engineered human pancreatic cells expressing shRNA to downregulate the glutamine transporter SLC1A5. We performed high-resolution 1H magnetic resonance spectroscopy (MRS) to understand the metabolic reprograming that occurs with SLC1A5 downregulation in these two cell lines. Method: Panc1 and Pa04C pancreatic cancer cell lines were genetically engineered to express shRNA against SLC1A5 using lentiviral transduction. Downregulation of SLC1A5 was verified with qRTPCR and immunoblotting. Control cells expressing an empty vector (EV) were also engineered. Dual phase extraction was performed as previously described [3]. High-resolution 1H MRS was performed on the aqueous phase extracts and recorded on a 750 MHz spectrometer. All data acquisition, processing and quantification was performed with TOPSPIN 3.5 software. Spectra were obtained from Panc1EV (n=4), Pa04CEV (n=4), Panc1_SLC1A5_shRNA (n=4) and Pa04C_SLC1A5_shRNA (n=4) cells. Results and Discussion: SLC1A5 downregulation significantly decreased the glutamine/glutamate ratio in Pa04C_SLC1A5_shRNA cells but not in Panc1_SLC1A5_shRNA cells. Instead, in Panc1_SLC1A5_shRNA cells, a significant decrease of the amino acids leucine, isoleucine, valine, tyrosine, histidine, and phenylalanine was observed. Lactate and fumarate also significantly decreased in Panc1_SLC1A5_shRNA cells. Other than the significant decrease of glutamine/glutamate in the Pa04C_SLC1A5_shRNA cells, no other significant metabolic differences were observed in these cells. Our data expand the understanding of the diverse metabolic reprogramming that occurs following SLC1A5 downregulation in two human pancreatic cancer cell lines that may lead to the development of additional metabolic targets. Supported by NIH R01CA193365 and R35CA209960.1. Son J et al., Nature. 2013; 2. Bhutia YD et al., Biochim Biophys Acta. 2016; 3. Winnard PT Jr. et al., J Cachexia Sarcopenia Muscle. 2020. Citation Format: Raj Kumar Sharma, Balalji Krishnamachary, Ishwarya Sivakumar, Yelena Mironchik, Santosh Kumar Bharti, Zaver M. Bhujwalla. Metabolic reprogramming by SLC1A5 downregulation in pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2353.

Catalytic hydrothermal liquefaction of biomass into bio-oils and other value-added products – A review

Hydrothermal liquefaction (HTL) is an effective method for the conversion of biomass into biofuel involving moderate temperature and high pressure. However, the yield and quality of bio-oil are not sufficient for commercialization under normal HTL conditions. In this regard, the review emphasizes the need to select the appropriate catalyst for achieving high bio-oil yields with improved quality. Besides, the factors influencing the catalytic HTL, including the types of catalyst used in HTL, the mechanism of catalytic-HTL reaction, the effect of the catalyst on other HTL products including biochar, aqueous phase extracts, and gas, are discussed in detail. This review also summarizes by mentioning the barriers to be overcome in catalytic HTL by focusing on promising approaches.

Assessment of in vitro potency of inactivated Newcastle disease oil-adjuvanted vaccines using hemagglutination test and blocking ELISA.

Aim:The present study was aimed to establish a protocol for the evaluation of the in vitro potency of commercial inactivated Newcastle disease virus (NDV) oil-adjuvanted vaccines using hemagglutination test (HA) and blocking ELISA (B-ELISA) based on polyclonal antibodies.Materials and Methods:Aqueous phases from a total of 47 batches of inactivated NDV vaccines manufactured by 20 different companies were extracted with isopropyl myristate. The viral antigen in each sample was detected and quantified by a standard HA test and a B-ELISA assay. To verify the efficiency of the antigen extraction method used in the batches which showed HA and to test the validity of using in vitro antigen quantification by HA and B-ELISA tests, a subset of 13 batches (selected from the total 47 batches) was inoculated in groups of 3-4-week-old specific pathogen-free chickens using the recommended vaccine dose. The immunogenicity of the selected vaccine batches was assessed by the NDV-hemagglutination inhibition antibody titers in individual serum samples collected 4 weeks after vaccination. Further, the efficacy of the vaccines and their protection rates were determined by a challenge test carried out for the vaccinated chickens with the Egyptian 2012 isolate of the virulent NDV genotype VII.Results:A strong correlation was observed between HA titers and B-ELISA mean titers in the tested 47 batches (R2=0.817). This indicated the possibility of using the latter in vitro assays for vaccine potency assessment. The recommended protective NDV antigen titer measured by B-ELISA was determined to be 28 ELISA units per dose. The comparison between the HA titers of the aqueous extracts of test vaccines and the corresponding results of in vivo potency assays (i.e., immunogenicity and efficacy), including antibody titers in the serum of vaccinated birds, indicated that the efficiency of the antigen extraction used may interfere with obtaining a strong correlation between the in vitro and in vivo results.Conclusion:HA or B-ELISA tests can be used as rapid and cost-effective alternatives to traditional in vivo potency tests for vaccine potency assessment by quantifying the NDV antigen present in aqueous phase extracts of the tested vaccines. The latter in vitro protocol, however, requires efficient extraction of the antigen to be able to obtain good correlation with the traditional in vivo potency tests.

Sulfate radical-initiated formation of isoprene-derived organosulfates in atmospheric aerosols

Recent studies show that isoprene-derived organosulfates are an important fraction of ambient secondary organic aerosol (SOA), adding up to 20% to the organic mass. Organosulfates with m/z of 199 and 183 relating to C4 compounds are found in ambient and laboratory generated SOA and a sulfate radical induced oxidation of methacrolein (MACR) and methyl vinyl ketone (MVK) has been shown to be a possible formation mechanism. In the present study, experiments on the sulfate radical-induced oxidation of methacrolein and methyl vinyl ketone were performed in bulk aqueous phase, as well as in an aerosol chamber, and finally compared with ambient PM10 samples collected at a rural East German village during the summer 2008, to investigate their relevance in aqueous phase SOA formation. Samples from aqueous phase experiments and extracts from filters were analysed with UPLC/(-)ESI-IMS-QTOFMS. All the samples showed the abundance of highly oxidised organosulfates with m/z 153, 155, 167, 183 and 199 corresponding to the species found in ambient particle samples. In the bulk phase studies with laser-induced sulfate radical formation, the signal intensities increased with increasing number of laser pulses, indicating the sulfate radical-induced formation of these organosulfates. Additionally, the chamber experiments showed a particle mass growth of about 10 microg m(-3) and 4 microg m(-3) for experiments on the reactive uptake of MACR and MVK with a sulfate radical precursor (K2S2O8) in the seed particles. Correlations of the C2 to C5 organosulfate species (including the m/z 215, C5H11O7S-), detected in the ambient samples were found to be very strong (r > 0.8), indicating that these compounds are formed from similar mechanisms and under equal environmental conditions. This study shows that sulfate radical-induced oxidation in the aqueous particle phase provides a reasonable explanation for the formation of these organosulfates from methacrolein and methyl vinyl ketone.

Effect of meconium on surface properties of surfactant monolayers and liposomes

The pathophysiology of meconium aspiration syndrome (MAS) is attributed in part to the development of lung surfactant dysfunction in vivo. Meconium (the first excrement of the newborn) and its components are responsible for development of lung surfactant dysfunction. In this study, the effect of meconium and its components on the surface activity of spread surfactant monolayers was evaluated using Wilhelmy balance at physiological temperature and pH. The effect of meconium on adsorption, airway patency and ultrastructure of liposomes made of various surfactant lipids was also evaluated. Significant inhibition of DPPC monolayer surface activity was observed on addition of meconium, its organic and aqueous phase extracts. All the surfactant lipid mixtures including DPPC attained significantly higher minimum surface tensions (between 12 and 20 mN/m) and lowered monolayer compressibility modulus in presence of meconium suggestive of surfactant monolayer fluidization. Meconium reduced the airway patency and affected the adsorption of DPPC liposomes and was associated with altered ultrastructure of DPPC liposomes. Meconium caused the maximum inhibitory effect on DPPC:PE and DPPC:PG and the least inhibitory effects on DPPC:POPC and DPPC:CHOL in terms of minimum surface tensions. All the surfactants were equally inhibited in terms of airway patency. These findings have implications for the differential inhibition of different surfactants in MAS and for the designing of surfactants that are resistant to meconium inhibition.

Nutritional Bioactives in Tropical Fruit

The nutritional benefits of tropical fruits are poorly understood. We assessed mango flavonoid and xanthone constituents (quercetin, mangiferin and its putative metabolite norathyriol) as well as whole extracts of mango peel and flesh on the proliferation of MCF‐7 breast cancer cells using an MTS assay. A significant decrease in cell proliferation was observed for the aqueous phase extracts from both peel and flesh. The compounds quercetin and norathyriol, also significantly inhibited MCF‐7 cell proliferation. We also assessed the effects of quercetin, mangiferin and norathyriol on human peroxisome proliferator‐activated receptors (PPARs), PPARα, PPARβ and PPARγ in Cos 7 cells. Norathyriol and quercetin dose‐dependently inhibited PPARs activated by their specific ligands. The IC50 value for quercetin was 59.6 μM for PPARα, 76.9 μM for PPARβ and 56.3 μM for PPARγ. For norathyriol, the IC50 value was 92.8 μM for PPARα, 102.4 μM for PPARβ and 153.5 μM for PPARγ. Mangiferin did not significantly inhibit PPAR transactivation (IC50 > 1 mM). Our results show that mangoes represent a potential source of bioactive compounds that have biological activity against the proliferation of cancer cells. Future studies will assess mangoes for other potential bioactive compounds and will assess other transcriptional pathways, such as the estrogen receptor pathway.

A review on advantages of implementing luminescence inhibition test (Vibrio fischeri) for acute toxicity prediction of chemicals

Evaluation of biological effects using a rapid, sensitive and cost effective method can indicate specific information on toxicity/ecotoxicity. Since assays based on animals, plants and algae are expensive, time consuming and require large sample volume, recent studies have emphasized the benefits of rapid, reproducible and cost effective bacterial assays for toxicity screening and assessment. This review focuses on a bacterial assay, i.e., Vibrio fischeri bioluminescence inhibition assay, which is often chosen as the first test in a test battery based on speed and cost consideration. The test protocol is simple and was originally applied for aqueous phase samples or extracts. The versatility of the assay has increased with subsequent modification, i.e., the kinetic assay for turbid and colored samples and the solid phase test for analyzing sediment toxicity. Researchers have reported the Vibrio fischeri bioluminescence assay as the most sensitive across a wide range of chemicals compared to other bacterial assays such as nitrification inhibition, respirometry, ATP luminescence and enzyme inhibition. This assay shows good correlations with other standard acute toxicity assays and is reported to detect toxicity across a wide spectrum of toxicants.

Characterization and mass load estimates of organic compounds in agricultural irrigation runoff

Investigations of agricultural chemicals in surface runoff typically target nutrients or specific pesticides; however, numerous other organic compounds are regularly applied to agricultural fields in pesticide formulations, irrigation water, soil amendments and fertilizers. Many of these compounds have toxicological significance. We conducted a broad spectrum analysis of surface runoff from individual irrigated agricultural fields in coastal southern California to characterize organic compounds amenable to analysis by gas chromatography-mass spectrometry and to estimate the mass flux of selected chemicals. Aqueous phase extracts contained several pesticides, as well as personal care product ingredients and pharmaceutically active compounds apparently derived from treated wastewater used for irrigation. Several compounds potentially associated with pesticide adjuvants were also present in aqueous phase extracts. Dissolved NOM constituents in water phase extracts included n-fatty acids, aliphatic alcohols and plant terpenoids. Tentatively identified compounds sorbed to suspended particles included pesticides, a fecal sterol, aliphatic and alicyclic hydrocarbons, aliphatic alcohols, aldehydes, and C14 and C16 n-fatty acids and fatty acid esters. Bicyclic and polycyclic aromatic hydrocarbons were identified in both aqueous and suspended particle phases. Constituent concentrations, including total suspended solids (TSS), varied over the course of the sampled events by up to an order of magnitude, and typically were not correlated with flow. Variation in sorbed organic compound concentrations often did not parallel those for TSS concentration. Mass load estimates were strongly influenced by the choice of sampling interval.

Lipo-oligosaccharide profiles of Serpulina pilosicoli strains and their serological cross-reactivities.

The purpose of this study was to determine the presence of lipopolysaccharide-like material in the intestinal spirochaete Serpulina pilosicoli and the extent of antigenic cross-reactivity of this material in different strains of the species. Hot water-phenol, aqueous-phase extracts from five porcine and three human strains of S. pilosicoli, and from seven strains of four other Serpulina spp., were separated by SDS-PAGE and silver-stained profiles were obtained. All S. pilosicoli strains had a predominant band at c. 16 kDa. Some also had a partial ladder-like profile, consistent with the presence of semi-rough lipo-oligosaccharide (LOS); this was more obvious in Western immunoblot analysis. LOS from each S. pilosicoli strain was serologically distinct in immunoblot analysis and there was no cross-reactivity with other Serpulina spp. The serological diversity found amongst the LOS of S. pilosicoli strains may help to explain why individual people and animals can suffer repeated infections with different strains of the organism.

Presence of proanthocyanidins in mutant green Sarracenia indicate blockage in late anthocyanin biosynthesis between leucocyanidin and pseudobase

The insectivorous pitcher plant genus Sarracenia contains several species with mutants which lack red pigment in leaves, flowers and growth point. Mutants and wild-type contain various glycosides of the flavonols quercetin and kaempferol and we hypothesized that the blockage in the mutant occurred between either dihydroflavonols and leucoanthocyanidin or between leucoanthocyanidin and the end product anthocyanin. Acid hydrolysis and acid addition were used to quantify and qualify spectral characteristics of intermediates. Aqueous phase extracts of leaves demonstrated the end product was most likely an anthocyanin because of red color, absorption at 530 nm and miscibility with amyl alcohol. Mutants and wild-type plants in Sarracenia purpurea and S. rubra ssp. jonesii are capable of producing proanthocyanidins while only wild-type contain pseudobase. Sarracenia purpurea mutants had a significantly greater accumulation of proanthocyanidins than wild-type while there was no such significant difference in S. rubra ssp. jonesii. These results suggest that the mutant is blocked in a late stage of anthocyanin biosynthesis between leucocyanidin and pseudobase. While mutants may accumulate precursors, additional investigation is needed. Sarracenia may serve as useful plants for research since the highly modified carnivorous and absorptive leaves may be fed suspected intermediates in this last step of anthocyanin synthesis.

Critical aspects of fluorescent age-pigment methodologies: Modification for accurate analysis and age assessments in aquatic organisms

Fluorescent age-pigment (FAP) quantification has recently been proposed as a means of estimating age in aquatic vertebrates and invertebrates. However, various aspects related to currently adopted procedures remain untested and a distinct relationship between FAP accumulation and chronological age has yet to be established. The present study was undertaken to critically evaluate the effects of specimen and extract handling procedures (including temperature, time, ultrasonication, and solvent systems) on the expression of native FAP fluorescence using the post-mitotic tissues of laboratoryreared specimens of the teleostOreochromis mossambicus as model systems. FAP-like fluorophores increased in vitro in brain, heart, and muscle tissues and their extracts with increased storage temperature (−20°C and above) and time. Sonification of homogenates greatly enhanced this effect and generated other non-native fluorophores in sample solution. Fluorescence assay temperature also affected expression of results, and aqueous-phase extracts, used in previous studies, were found to contain large amounts of fluorescent flavin contaminants. Using modifications of the above procedures, age-related patterns of accumulation were subsequently examined inO. mossambicus brain and, for the first time, positive correlations between chronological age and whole organ as well as weight-specific FAP concentration were validated. The significance of these findings are discussed in relation to previous FAP studies.

The I, i, and HI blood group antigens in extracts of human erythrocytes.

Extracts of stromata of human adult and newborn (cord) erythrocytes were prepared with n-butanol. The aqueous phase and butanol phase extracts were examined for the presence of blood group I, i, HI and H substances in inhibition of agglutination experiments. The recovery of HI activity in both the aqueous and butanol phase extracts of adult red cells but only in the aqueous phase of cord cells is reported heare for the first time. The i specificity was present in the aqueous phase but not in the butanol phase of cord erythrocytes, also not previously reported. Although the recovery of I but not i substance in aqueous phase extracts of adult cells had been shown by other workers, examination of the butanol phase extracts for I and i substances had not been documented. In the present study, Ii substances were not demonstrable in any of the butanol phase extracts of adult and cord erythrocytes.

The A, M, N, and P1 blood group substances in butanol extracts of human erythrocytes.

Aqueous phase and butanol phase extracts of group A1, O, M, N, P1 and P2 human erythrocytes perpared by extraction of red cell stromata with n-butanol were examined for the presence of blood group substances in inhibition of agglutination experiments using antisera and test cells of human origin. Group A substance was recovered in both aqueous phase and butanol phase extracts, M and N substance in aqueous phase extracts only, and P1 substance only in butanol phase extracts. Use of high concentrations of extract resulted in detection of P1 activity in aqueous phase extracts and of N specific inhibition in the aqueous phase extracts of group M erythrocytes. Distilled water appeared to be superior to digitonin and hypotonic phosphate buffer solutions for preparation of stromata for the recovery of group A and M substances in the butanol extracts but not for group N and P1 substances.

The H, HI, I and i blood group antigens in butanol extracts of rabbit tissues.

Aqueous and butanol phase extracts of rabbit erythrocytes and the tissues gastric mucosa, liver, lung and kidney were examined by inhibition of agglutination for the presence of H, HI, I and i blood group substances. Of the four specificities, only I substance was recovered from erythrocytes and only in the aqueous phase extracts. Both aqueous and butanol extracts of gastric mucosae contained H substance, but HI and I were present only in the aqueous extracts of this tissue. Extracts of rabbit lungs only had H and I activity and only in the aqueous phase. No H, HI, I or i substance was detected in any of the extracts of liver and kidney. Reduced levels of H substance were found in aqueous extracts of gastric mucosae of 'A-like' rabbits. Three H specificities were demonstrated in the butanol phase extracts of gastric mucosae of rabbits. These specificities were similar to those previously shown in extracts of gastric mucosae and erythrocytes of humans.

The H specificities of butanol extracts of human erythrocytes.

A previous report of other researchers that aqueous phase fractions of erythrocyte stromata of secretors contained more ABH blood group activity than similar fractions of non-secretors was confirmed. The recovery of H activity in the butanol phase extracts of both secretors and non-secretors, which had not been seen previously, is reported here for the first time. Three H specificities were present when the aqueous and butanol phases of group O secretor stromata were examined in inhibition of agglutination studies with anti-H of group Oh and group A1 sera and three lectins. The anti-H (group Oh) sera were inhibited by both aqueous and butanol fractions. Anti-H serum (group A1) and the lectin Ulex europaeus were inhibited by the aqueous fraction only. The lectins Laburnum alpinum and Lotus tetragonolobus were not inhibited by either the aqueous or butanol phase extracts.

The H Specificities of Butanol Extracts of Human Erythrocytes

A previous report of other researchers that aqueous phase fractions of erythrocyte stromata of secretors contained more ABH blood group activity than similar fractions of non-secretors was confirmed. The recovery of H activity in the butanol phase extracts of both secretors and non-secretors, which had not been seen previously, is reported here for the first time. Three H specificities were present when the aqueous and butanol phases of group O secretor stromata were examined in inhibition of agglutination studies with anti-H of group O(h) and group A(1) sera and three lectins. The anti-H (group O(h)) sera were inhibited by both aqueous and butanol fractions. Anti-H serum (group A(1)) and the lectin Ulex europaeus were inhibited by the aqueous fraction only. The lectins Laburnum alpinum and Lotus tetragonolobus were not inhibited by either the aqueous or butanol phase extracts.