Allergy to a garlic lectin in an infant after the first intake: difficulties identifying the route of sensitization

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Formulation of chitosan patch incorporating Artocarpus altilis heartwood extract for improving hyperpigmentation.

Artocarpus altilis heartwood extract contains the bioactive compound artocarpin which exhibits melanogenesis inhibitory activity. However, the extract has poor solubility which affects the skin permeability of the compound. A chitosan hydrogel patch incorporating A. altilis heartwood extract was formulated to enhance the delivery of an amount of artocarpin sufficient for depigmenting the skin. The extract was prepared as an o/w microemulsion before blending with an aqueous solution of chitosan. The hydrogel patch was formulated by blending in a 1:1 ratio by weight of 4% w/w chitosan solution and 0.04% w/w extract microemulsion which provides optimal values of the mechanical properties of the patch. The release of artocarpin from the formulated patch (artocarpin content, 0.07 mg/cm2) exhibited two phases; the rapid rate (0-15 min) averaged 0.73 µg/min/mm2, and the slow rate (15-240 min) averaged 0.02 µg/min/mm2. The formulated patches significantly improved the hyperpigmented area of the subjects after 3 weeks of application. No adverse events were observed. The results indicate that the formulated chitosan hydrogel patch delivers an effective amount of incorporated artocarpin depigmenting action.

Studies in a Murine Model Confirm the Safety of Griffithsin and Advocate Its Further Development as a Microbicide Targeting HIV-1 and Other Enveloped Viruses.

Griffithsin (GRFT), a lectin from Griffithsia species, inhibits human immunodeficiency virus-1 (HIV-1) replication at sub-nanomolar concentrations, with limited cellular toxicity. However, in vivo safety of GRFT is not fully understood, especially following parenteral administration. We first assessed GRFT's effects in vitro, on mouse peripheral blood mononuclear cell (mPBMC) viability, mitogenicity, and activation using flow-cytometry, as well as cytokine secretion through enzyme-linked immunosorbent assay (ELISA). Toxicological properties of GRFT were determined after a single subcutaneous administration of 50 mg/kg or 14 daily doses of 10 mg/kg in BALB/c mice. In the context of microbicide development, toxicity of GRFT at 2 mg/kg was determined after subcutaneous, intravaginal, and intraperitoneal administrations, respectively. Interestingly, GRFT caused no significant cell death, mitogenicity, activation, or cytokine release in mPBMCs, validating the usefulness of a mouse model. An excellent safety profile for GRFT was obtained in vivo: no overt changes were observed in animal fitness, blood chemistry or CBC parameters. Following GRFT treatment, reversible splenomegaly was observed with activation of certain spleen B and T cells. However, spleen tissues were not pathologically altered by GRFT (either with a single high dose or chronic doses). Finally, no detectable toxicity was found after mucosal or systemic treatment with 2 mg/kg GRFT, which should be further developed as a microbicide for HIV prevention.

The "white kidney bean incident" in Japan.

Lectin poisoning occurred in Japan in 2006 after a TV broadcast that introduced a new diet of eating staple foods with powdered toasted white kidney beans, seeds of Phaseolus vulgaris. Although the method is based on the action of a heat-stable α-amylase inhibitor in the beans, phaseolamin, more than 1,000 viewers who tried the method suffered from acute intestinal symptoms and 100 people were hospitalized. Lectins in the white kidney beans were suspected to be the cause of the trouble. We were asked to investigate the lectin activity remaining in the beans after the heat treatment recommended on the TV program. The test suggested that the heat treatment was insufficient to inactivate the lectin activity, which, combined with our ignorance of carbohydrate signaling in the intestine, was the cause of the problem.

A lectin from the green seaweed Caulerpa cupressoides reduces mechanical hyper-nociception and inflammation in the rat temporomandibular joint during zymosan-induced arthritis.

Seaweed lectins have been widely investigated as anti-nociceptive and anti-inflammatory agents. This study analyzed the anti-nociceptive and anti-inflammatory responses of a lectin from the green seaweed Caulerpa cupressoides (CcL) on zymosan-induced arthritis of the rat temporomandibular joint (TMJ). Rats received i.v. CcL 30 min prior to injection of zymosan (2mg/art.) or 0.9% saline into the left TMJ. Mechanical hyper-nociception was measured by the electronic von Frey method at baseline and 4h after zymosan injection. Animals were euthanized 6h after zymosan injection and the synovial fluid was collected for leukocyte counting and myeloperoxidase activity assessment. Other animals were treated with ZnPP-IX (3mg/kg; s.c.), a specific heme oxygenase-1 pathway inhibitor, and naloxone (10 µg/art.), a nonselective opioid receptor antagonist. TMJ tissues were excised to perform histopathological and immunohistochemistry analyses. CcL (0.1, 1 or 10mg/kg) significantly reduced zymosan-induced hyper-nociception (81, 83 and 89.5%, respectively) and inhibited the leukocyte influx (77.3, 80.7 and 98.5%, respectively) compared with the zymosan-only group, as confirmed by myeloperoxidase activity; however, treatment with naloxone or ZnPP-IX did not revert the effects of CcL (10mg/kg), suggesting that the naloxone-sensitive opioid and heme oxygenase-1 pathways are not involved. CcL also reduced the leukocyte influx and the expression of IL-1ß and TNF-α in the TMJ, based on histopathological and immunohistochemistry analyses, respectively. Therefore, CcL reduces TMJ hyper-nociception and inflammation with a mechanism that is partially dependent on TNF-α and IL-1ß inhibition. CcL reveals a potentially valuable alternative tool for future studies of TMJ disorders.

Component-resolved immunologic modifications, efficacy, and tolerance of latex sublingual immunotherapy in children.

BACKGROUND: As the frequency of natural rubber latex (NRL) allergy has increased, attempts have been made to diminish exposure in high-risk patients. Despite some good results, complete NRL avoidance was not possible, so latex immunotherapy was developed. OBJECTIVE: To examine variations in immunologic parameters, clinical efficacy, and safety of NRL sublingual immunotherapy (SLIT). METHODS: This prospective, observational, open, case-control study included 23 patients (18 patients receiving NRL SLIT and 5 controls). Skin prick, conjunctival provocation, and in-use tests with NRL, specific IgE and specific IgG4 to NRL, specific IgE to recombinant NRL allergens, and basophil activation test (BAT) with whole latex, natural, and recombinant allergens were performed before immunotherapy (T0) and at 6 (T1) and 12 months (T2) of treatment. RESULTS: Patients were sensitized to Hev b 5, Hev b 6.01, and Hev b 6.02 proteins, optimal for SLIT. Changes in specific IgE were not significant. Increases in specific IgG4 between T1 and T2 were larger in the active group. BAT determinations showed significant decreases in recombinant Hev b 6.01 and natural Hev b 6.02 in the active group at T1 but not at T2. Both groups had new sensitizations at T1 but not at T2. The active group had significant increases in the response threshold in the in vivo tests at T1 and T2. Adverse effects were limited to local reactions. CONCLUSION: NRL SLIT is effective and safe in children with latex allergy. Our results suggest that specific IgE determinations and BAT measurements to natural and recombinant latex allergens may allow obtaining an allergen-based diagnosis to help determine specific immunotherapy.

Effects of Lens culinaris agglutinin on gene expression of gluconeogenic enzymes in the mouse intestine.

BACKGROUND: Lectins are proteins that bind specifically to the carbohydrate moiety of glyco-conjugates. Japanese mistletoe lectin given intragastrically affected cytokine gene expression in the mouse intestine. This study examines the actions of Lens culinaris agglutinin (LCA) on the gene expression of gluconeogenic enzymes in the intestine. RESULTS: The results of quantitative real-time reverse transcription-polymerase chain reaction indicated that LCA caused an up-regulation of the gene expression of glucose-6-phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK). This change was correlated with an increase in the expression of two transcription factors, HNF1α and HNF4α. Experiments using human colonic cancer Caco-2 cells demonstrated that LCA up-regulated the gene expression of G6Pase and PEPCK whereas insulin had the opposite effect. In addition, the observed up-regulation of HNF4α gene expression in the duodenum raises the possibility that the lectin promotes the colorectal cancer. CONCLUSION: Lentil beans should be cooked well to avoid unfavourable effects of LCA.

Protective effect of Artin M from extract of Artocarpus integrifolia seeds by Th1 and Th17 immune response on the course of infection by Candida albicans.

The immunoregulatory effect of Artin M and jacalin from extract of Artocarpus integrifolia seeds (jack extract) against infection with Candida albicans was investigated. Swiss mice received jack extract containing 500 µg protein/ml PBS intraperitoneally (i.p.) or PBS alone and after 72 h were infected i.p. with C. albicans CR15 (10(7)) and sacrificed after 30 min, 2, 6, 24, and 72 h. ELISA analysis revealed that in jack extract-treated mice IFN-γ was predominantly produced versus IL-10 in control mice. These results suggest that jack extract induced a protective immune response, since C. albicans clearance was complete at 72 h postinfection. Jack extract presents two lectins (Artin M and jacalin) with distinct biological properties. Artin M was able to induce IL-12 production by macrophages. Also, Artin M in different concentrations, associated with jacalin or in jack extract induced both IFN-γ and IL-17 production. As a consequence, phagocytic and candidacidal activity increased significantly. Alanine aminotransferase activity (ALT) was used as parameter for damage of the liver. The activity of ALT correlated with inoculum size that increased significantly in control group, however, mice pretreated with jack extract 3 days before infection presented normal ALT. Mice pretreated with jack extract that received a lethal inoculum of Candida presented 90% survival versus 20% among controls or mice pretreated with jacalin. Thus, the results suggest that Artin M by itself, associated with jacalin or present in jack extract is able to induce protective Th1 and Th17 immune responses against Candida albicans infection.

Reduced side-effects of adjuvant hormone therapy in breast cancer patients by complementary medicine.

UNLABELLED: A clinical investigation (representing evidence-based medicine level III) was performed to evaluate the benefit of complementary medicine in breast cancer patients undergoing adjuvant hormone therapy (HT). PATIENTS AND METHODS: The patients (n=129) were treated according to international guidelines. All patients suffered from arthralgia and mucosal dryness induced by the adjuvant HT. To reduce these side-effects, the patients were complementarily treated with a combination of sodium selenite, proteolytic plant enzymes (bromelaine and papain) and Lens culinaris lectin. On the basis of case report formulas (CRFs), self assessment of defined side-effects of HT (arthralgia and mucosal dryness) were documented before as well as 4 and 8 weeks after complementary treatment. Validation was carried out by scoring from 1 (no side-effects/optimal tolerability) to 6 (extreme side-effects/extremely bad tolerability). RESULTS: The severity of side-effects of HT was reduced by complementary treatment with sodium selenite, plant enzymes (bromelaine and papain) and Lens culinaris lectin. The mean score of symptoms declined from 4.2 (before treatment) to 3.2 (after 4 weeks of treatment) to 2.7 (after 8 weeks of treatment) for arthralgia and from 3.2 (before treatment) to 2.9 (after 4 weeks of treatment) to 2.6 (after 8 weeks of treatment) for mucosal dryness, the primary aims of this investigation. The reduction of side-effects of HT was statistically significant (p<0.001 after 4 weeks and p<0.0001 after 8 weeks). CONCLUSION: This investigation demonstrates benefits of indication-based complementary treatment in breast cancer patients, e.g. reduction of side-effects of adjuvant HT. A randomized controlled trial is planned to integrate the complementary treatment with the combination of sodium selenite, proteolytic enzymes and Lens culinaris lectin into evidence-based medicine.

Effect of feeding a formula supplemented with long-chain polyunsaturated fatty acids for 14 weeks improves the ex vivo response to a mitogen and reduces the response to a soy protein in infants at low risk for allergy.

BACKGROUND AND OBJECTIVE: Feeding long-chain polyunsaturated fatty acids (LCP) influences immunity in adults; however, less is known about their effect during development. The aim of the study was to determine the effect of feeding LCP on immunity in healthy infants during the first 4 months of life. PATIENTS AND METHODS: Formula-fed infants were randomized at

The nutraceutical role of the Phaseolus vulgaris alpha-amylase inhibitor.

The present review assesses the potential of the Phaseolus vulgaris alpha-amylase inhibitor isoform 1 (alpha-AI1) starch blockers as a widely used remedy against obesity and diabetes. Consumption of the alpha-amylase inhibitor causes marginal intraluminal alpha-amylase activity facilitated by the inhibitor's appropriate structural, physico-chemical and functional properties. As a result there is decreased postprandial plasma hyperglycaemia and insulin levels, increased resistance of starch to digestion and increased activity of colorectal bacteria. The efficacy and safety of the amylase inhibitor extracts, however, depend on the processing and extraction techniques used. The extracts are potential ingredients in foods for increased carbohydrate tolerance in diabetics, decreased energy intake for reducing obesity and for increased resistant starch. Research developments in the distribution and biosynthesis of the alpha-amylase inhibitor, relevant physico-chemical properties, the molecular starch-blocking mechanism, anti-obesity and anti-diabetes effects, safety of extracts and the need for research into their potential anti-colorectal cancer effect are discussed.

Differential expression of allergens on the internal and external surfaces of latex surgical gloves.

BACKGROUND: Differences in latex allergen sensitization profiles have been described between children undergoing repeated surgical interventions and health care workers. The purpose of this study was to determine whether such sensitization profiles are associated with differences in the expression of latex allergen between the internal and external surfaces of surgical gloves. METHODS: Extracts were obtained from whole surgical gloves as well as from their external and internal surfaces. The extracts were centrifuged, filtered, concentrated, dialyzed and lyophilized. The protein profile of the extracts was analyzed using hydrophobic interaction chromatography (HIC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting was performed using sera from two patients with confirmed latex allergy. Latex recombinant allergen-specific IgE in these two patients was determined using a fluorescence enzyme immunoassay (FEIA) method. Latex allergen quantification was determined on both glove surfaces using an ELISA method. RESULTS: HIC and SDS-PAGE showed qualitative and quantitative differences in proteins between the internal and external glove surfaces, with the former being much richer in proteins. Immunoblotting of glove extracts using sera from two latex-allergic health workers showed differences between glove surface extracts. ELISA quantification of latex allergens demonstrated that the internal glove surface had high amounts of Hev b 5 and Hev b 6.02 whereas the external surface showed Hev b 1, Hev b 3, and Hev b 6.02. CONCLUSIONS: Our results reveal substantial differences in the composition of latex allergen profiles between the internal and external surfaces of surgical latex gloves, which may suggest a relationship between latex allergen localization and sensitization routes in different risk groups.

Effect of horse gram lectin (Dolichos biflorus agglutinin) on degranulation of mast cells and basophils of atopic subjects: identification as an allergen.

Horse gram (Dolichos biflorus) is widely consumed in the tropical south Asian countries including rural areas of India. Since D. biflorus agglutinin (DBA) is an important dietary lectin in horse gram, we have studied its effect on the degranulation of mast cells and basophils of atopic subjects. Allergy to horse gram lectin has not been reported so far. Skin prick test (SPT) was performed with 100 microg/mL of DBA. DBA-specific IgE was detected by dot-blot, and ELISA. Histamine release (HR) assay was carried out using leukocytes from non-atopic and atopic subjects, and rat peritoneal exudate cells. Among the atopic group, 10 of 48 subjects (21%) were found to be positive for DBA by SPT, and none were positive in the non-atopic group (n=20). Two subjects out of the ten who tested positive for DBA by SPT were found to be sensitized to DBA as revealed by the presence of specific IgE by ELISA and dot-blot. The HR was found to be 2- to 3-fold higher in DBA-allergic subjects than in non-atopic and atopic subjects. Basophil HR by DBA was found to be similar in both non-atopic and atopic subjects. However, DBA induces activation of mast cells in vivo in a sub-population (21%) of atopic subjects. Two subjects have been identified as having food allergy to horse gram based on the presence of DBA-specific IgE with a positive correlation to basophil HR. This is the first report of food allergy to horse gram, and DBA has been identified as an allergen.

The effect of raw full-fat soybean and its lectin on the nutrition and pigmentation of broilers.

This study was aimed at determining the effect of raw full-fat soybean in the digestion/absorption of broilers and to evaluate the role of soybean agglutinin (SBA) in the pathogenesis of the runting and stunting syndrome of broilers. Six broiler groups were fed with six experimental diets for 42 days in which the defatted soybean meal of the basal diet was substituted with increasing raw full-fat soybean percentages ranging from 0 to 100%. The results suggested that SBA included in the raw full-fat soybean can play a role in the pathogenesis of the runting and stunting syndrome in broilers since binding and incorporation of SBA to enterocytes were associated with hyperplasia and dysplasia of the intestinal epithelium, as suggested by the progressive increases (P < 0.001) of mitosis/crypt in the duodenum and in the midintestine, as well as by intestinal villi atrophy (P < 0.001) in the duodenum and the midintestine. Indigestion and malabsorption of proteins induced progressive low weight gain (P < 0.001) up to 50% with 100% of raw full-fat soybean in the diet at 42 days. Xanthophylls in serum and skin yellowness were enhanced linearly in the broilers fed 42 days with increasing percentages of raw full-fat soybean in the diet. Increased xanthophylls absorption and decreased protein absorption can be caused by pathological changes in the epithelial intestine triggered by SBA binding and internalization.

Soybean: feed quality and safety. Part 2: pathology of soybean feeding in animals. A review.

Various animal models in farm and laboratory animals indicate the possible pathological effect of soybean feeding. Immunity and tolerance to oral soybean intake and a generalised meta-model of effects are discussed. It seems that FAO/WHO recommendations will have to be considered in assessing the biological value of soybean feed sources. Determination of hull content, antigenic potential, conglycinin and trypsin inhibitor content is warranted in view of the future component pricing of soybean meals.

Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat.

This study was conducted to investigate the effects of a moderate dose of purified soybean agglutinin on performance and nitrogen digestibility in rats as well as to determine its effects on the protein, DNA and RNA content of the small intestine and pancreas. Twenty-four Sprague-Dawley rats were randomly allotted into one of four groups for a 10-day nitrogen balance experiment. The four groups of rats were fed 7 g of a casein-cornstarch based diet or a similar diet supplemented with 0.1, 0.2 or 0.4 mg/g purified soybean agglutinin. All experimental diets were adjusted to an identical nutrient level. Dose of soybean agglutinin had no significant effect on rat performance. Incorporation of soybean agglutinin in the diet reduced apparent protein digestibility and the utilization of dietary protein by increasing nitrogen loss from the faeces and urine. Fresh pancreatic weight increased in rats fed soybean agglutinin at a level of 0.4 mg/g in the diet compared to the control, but the dry pancreatic weight and the protein content of the pancreas did not differ among the four groups. However the DNA and RNA content of the pancreas had a tendency to increase with a higher level of soybean agglutinin. The weight of the jejunum and its protein, DNA and RNA content were not significantly affected by soybean agglutinin, but the dry weight and the RNA of the jejunum tended to increase with higher levels of soybean agglutinin in the diet. In conclusion, purified soybean agglutinin, at moderate levels in the rats diet, had negative effects on digestive function, such as nitrogen digestibility, nitrogen retention and nitrogen balance. As the level of soybean agglutinin increased, the effects became more pronounced. Meanwhile, hypertrophy of the pancreas was observed with higher doses of soybean agglutinin incorporation in the diets.