Showing papers in "Food and Agricultural Immunology in 2015"
TL;DR: This semi-quantitative detection method with high sensitivity and satisfactory recovery shows significant potential for CEX residue detection in milk.
Abstract: An anti-cephalexin (CEX) monoclonal antibody was prepared by the immunogen of CEX-keyhole limpet hemocyanin connected by disuccinimidyl suberate, which was used for the establishment of a rapid immunochromatographic test strip. With this semi-quantitative detection method, a low limit of deduction of 10 ng mL−1 was obtained with the naked eyes and 1.3 ± 0.1 ng mL−1 by a strip reader in unprocessed milk within 5 min, which was much lower than the European Union Maximum Residue Limit of 100 µg kg−1 in milk. The recovery in the range of 87–120% was measured in another milk sample. In conclusion, this method with high sensitivity and satisfactory recovery shows significant potential for CEX residue detection in milk.
59 citations
TL;DR: The results indicated that SRS helps the body prevent and treat obesity through mechanisms including synthesis and secretion of leptin (LP) and adiponcetin (ADP) and improvement in intestinal flora.
Abstract: Resistant starch, as a source of energy for the beneficial bacteria in intestine, may have beneficial health effects, limiting your risk for obesity and colon problems. This study evaluated the effects of sorghum resistant starch (SRS) on the changes of body weight, blood lipid and the population of intestinal flora in the colon of high-fat diet-induced (HFD) overweight and obese rats which contained 60 rats. Sixty male rats were divided into five groups of normal control group (NC), model control group (MC A), model control group (MC B), sorghum resistant starch A group (SRS A) and sorghum resistant starch B group (SRS B) which administrated standard diet, diets A and B. The results indicated that SRS helps the body prevent and treat obesity through mechanisms including synthesis and secretion of leptin (LP) and adiponcetin (ADP) and improvement in intestinal flora.
48 citations
TL;DR: Results indicated that this developed enzyme-linked immunosorbent assay (ELISA) is useful for the detection of carbendazim residues in fruits and vegetables.
Abstract: In the present study, a class-selective monoclonal antibody (mAb) for carbamate fungicides was obtained using two haptens. One hapten was designed as immunogen and the other one for coating. Under the optimum assay conditions of coating antigens H2-ovalbumin, in the assay buffer of sodium chloride 1.6%, no methanol, and pH 6.5, the assay system was more sensitive. The linear range was from 0.06 to 7.47 ng/mL. The cross-reactivity test was carried out with IC50 values of 0.45 ng/mL for carbendazim, 0.57 ng/mL for benomyl, and 9.70 ng/mL for thiabendazole. Other related compounds thiophanate methyl, fenbendazole, mebendazole, methyl paraoxon, methyl parathion, chlorpyrifos, and chlorothalonil did not cross-react significantly in this assay. To evaluate the proposed immunoassay, real samples such as apples, tomatoes, and cucumbers were subjected to recovery test with carbendazim. The recovery and coefficient variation range from 73% to 125% and 0.36% to 11.80%, respectively. The developed method was also com...
46 citations
TL;DR: This review focuses on seafood allergens, reporting an updated and compiled list of allergens from fish, crustaceans and mollusc species, with an overview on the most representative analytical methods for their detection.
Abstract: Food-induced allergies are considered an important problem of public health with special impact in the quality of life of the sensitised/allergic individuals. As highly consumed foods, fish and shellfish represent a valuable source of proteins for the general population. In spite of their economical and nutritional importance, these foods are known to induce hypersensitivity reactions in sensitised/allergic individuals. So far, parvalbumins (fish) and tropomyosins (crustaceans and molluscs) have been considered major allergens in seafood allergy, being responsible for most of the reported cases of adverse immunological responses. More recently, other proteins such as arginine kinases, myosin light chains, troponins and sarcoplasmic calcium-binding proteins have been regarded as relevant allergens in fish, crustaceans and molluscs. This review focuses on seafood allergens, reporting an updated and compiled list of allergens from fish, crustaceans and mollusc species, with an overview on the most representa...
41 citations
TL;DR: The residues of the antibiotics in trout samples did not exceed the maximum residue levels recommended by the Institute of Standards and Industrial Research of Iran, however, the high occurrence of tetracyclines and florfenicol in the samples is alarming and could be a potential hazard for public health.
Abstract: This study was carried out to determine the occurrence of tetracyclines, sulfonamides, fluoroquinolones and florfenicol residues in rainbow trout muscle samples (n = 138) obtained from Iranian trout farms Concentrations of the antibiotics were determined using competitive enzyme-linked immunosorbent assay method At measurable levels, 631%, 167%, 196% and 406% of the samples contained the residues of tetracyclines, sulfonamides, fluoroquinolones and florfenicol, respectively The detected range of concentrations for positive samples was 143–1014 µg/kg for tetracyclines, 403–904 µg/kg for sulfonamides, 675–998 µg/kg for fluoroquinolones and 011–1726 µg/kg for florfenicol The residues of the antibiotics in trout samples did not exceed the maximum residue levels recommended by the Institute of Standards and Industrial Research of Iran However, the high occurrence of tetracyclines and florfenicol in the samples is alarming and could be a potential hazard for public health Further investigation
41 citations
TL;DR: In this paper, residues of tetracycline groups were quantified in 120 broiler chickens carcasses (including breast, liver and thigh tissues) by high performance liquid chromatography (HPLC) and also four-plate test (FPT) and ELISA methods as primary screening tests.
Abstract: Residues of tetracycline (TC) groups were quantified in 120 broiler chickens carcasses (including breast, liver and thigh tissues) by high performance liquid chromatography (HPLC) and also four-plate test (FPT) and ELISA methods as primary screening tests. Results of FPT showed that 25% of carcasses were polluted by antibiotic residues. The ELISA assay indicated that out of 30 positive samples in FPT, 21 liver, 16 thigh and 12 breast muscle samples were positive for TC residues. Frequency of TC-like residues and their concentration ranges in ELISA-positive samples were determined by HPLC method. In comparison between the results of these methods, the correlation between FPT and HPLC (0.940) was higher than ELISA and HPLC methods (–0.726). Moreover, this study revealed that an inhibition test with a medium at pH 6 which seeded with Bacillus subtilis had highest sensitivity for detecting of TC residues.
39 citations
TL;DR: In this article, a convenient and specific competitive enzyme-linked immunosorbent assay (ELISA) for nadifloxacin (NDF) was developed, which could be used as a screening method to detect and control the illegal content of NDF in food products.
Abstract: A convenient and specific competitive enzyme-linked immunosorbent assay (ELISA) for nadifloxacin (NDF) was developed. The modified 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) method was employed to synthesise the artificial antigen of NDF. The indirect competitive ELISA developed to detect NDF has a concentration at 50% inhibition (IC50) of 0.72 ng/mL and a limit of detection of 0.04 ng/mL. This ELISA kit could be used as a screening method to detect and control the illegal content of NDF in food products. In this study, the kit was applied to NDF detection in chicken muscle and produced a satisfactory result with recovery rates from samples spiked with NDF of 92% (10 ng/mL) and 101% (50 ng/mL).
36 citations
TL;DR: In this study, 288 milk samples were collected randomly from individual farms in Qazvin province from March to February 2012, Iran and found to contain Aflatoxin M1 at various levels; the presence of AFM1 was detected in a concentration ranging between 0.01 and 0.22 ppb.
Abstract: In this study, 288 milk samples were collected randomly from individual farms in Qazvin province from March to February 2012, Iran. Samples were analysed for Aflatoxin M1 (AFM1) by ELISA; also the Aflatoxin B1 (AFB1) contamination in animal feed was determined at the same time as the sampling of raw milk. One hundred and sixty-three samples (56.59%) were found to contain AFM1 at various levels; the presence of AFM1 was detected in a concentration ranging between 0.01 and 0.22 ppb. In 113 (69.32%) of the 163 milk samples, AFM1 levels were found to be higher than the maximum tolerable limit (0.05 ppb) accepted by the European union/Codex Alimentarius Commission. The mean AFM1 contamination levels in milk in summer and winter were 0.08 and 0.18 ppb, respectively. The AFB1 contamination level in the winter feed (2.27 ± 1.76) was higher than from summer (0.83 ± 0.60) (P < 0.05).
34 citations
TL;DR: In this paper, three commercial enzymes orientase, flavourzyme and alcalase 2.4L were used to hydrolyze tuna cooking juice, and the protein hydrolysates were purified, before being evaluated for the ability to induce secretary and cellular responses in murine peritoneal macrophages RAW264.7.
Abstract: To produce and identify anti-inflammatory peptides, three commercial enzymes orientase, flavourzyme, and alcalase 2.4L were used to hydrolyze tuna cooking juice, and the protein hydrolysates were purified, before being evaluated for the ability to induce secretary and cellular responses in murine peritoneal macrophages RAW264.7. Alcalase hydrolysate (AH) exhibited the most potent inhibitory effect. AH was further purified using gel-filtration chromatography and high-performance liquid chromatography. The results showed that the peptide fractions with the molecular weight ranging from 204 to 1672.9 Da possessed the greatest anti-inflammatory activity. The amino acid sequences of the two anti-inflammatory peptides isolated from AH hydrolysates were Pro-Arg-Arg-Thr-Arg-Met-Met-Asn-Gly-Gly-Arg (1543.8 Da) and Met-Gly-Pro-Ala-Met-Met-Arg-Thr-Met-Pro-Gly (1211.5 Da). We thus conclude that anti-inflammatory hydrolysates from tuna cooking juice by-product may be useful ingredients in food and nutraceutical applic...
33 citations
TL;DR: The data suggested that immune stress can negatively affect the utilisation of energy in broilers, resulting in the decreased growth performance.
Abstract: This study was conducted to investigate the effects of different types of immune stress on the growth performance and energy metabolism in broiler chickens (Cobb 500). Three hundred and twenty one-day-old broilers were randomly assigned (n = 80 birds/group) to receive one of four treatments: (1) no vaccination, (2) conventional vaccination, (3) cyclophosphamide (CPM) stress and (4) lipopolysaccharide (LPS) stress. The results demonstrated that immune stress reduced the daily feed intake, average daily weight gain, plasma renin activity, serum angiotensin II and amylase activity of jejunal digesta (P < 0.05). The body temperature of broilers increased rapidly after LPS administration on days 16, 18 and 20, but attenuated on days 37, 39 and 41. The data suggested that immune stress can negatively affect the utilisation of energy in broilers, resulting in the decreased growth performance.
30 citations
TL;DR: A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established to detect the Cronobacter sakazakii.
Abstract: A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established to detect the Cronobacter sakazakii. A pair of monoclonal antibodies (mAbs) selected from mAbs produced by 11 murine hybridomas was selected for the sandwich ELISA procedure. Targets of two mAbs were 100 kDa and 42 kDa protein extracted from the bacteria, respectively, which were proved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting analysis. The limit of detection of this method was established as 1 × 104 cfu/mL, and the linear range from 1 × 105 to 1 × 108 cfu/mL. In the real sample test, 1 cfu/g C. sakazakii was detected in artificially contaminated powdered infant formula with 4 h enrichment.
TL;DR: Findings indicate that EPS and Se-EPS can enhance the immune response via enhanced macrophage and spleen lymphocyte stimulation and show stronger immunomodulatory activity than EPS.
Abstract: Exopolysaccharide (EPS) was isolated and purified from Lactococcus lactis subsp. Lactis culture and then selenium chloride oxide (SeCl2O) was added to synthesise selenium-exopolysaccharide (Se-EPS). This study sought to evaluate the immunomodulatory effects of EPS and Se-EPS on the activity of peritoneal macrophages and spleen lymphocytes. In mouse peritoneal macrophages, the EPS and Se-EPS enhanced phagocytosis and increased nitric oxide (NO), IFN-γ, IL-1, IL-6 and IL-12 secretion, but not IL-10 levels. Preliminary in vitro tests revealed that EPS and Se-EPS stimulated mouse spleen lymphocyte proliferation, and strongly improved levels of TNF-α, IL-2 and IL-6 mRNA in spleen cells. These findings indicate that EPS and Se-EPS can enhance the immune response via enhanced macrophage and spleen lymphocyte stimulation. Se-EPS showed stronger immunomodulatory activity than EPS.
TL;DR: A portable immunochromatographic strip based on gold-labelled monoclonal antibodies was developed for the rapid detection of Pseudomonas syringae pv.
Abstract: A portable immunochromatographic (IC) strip based on gold-labelled monoclonal antibodies was developed for the rapid detection of Pseudomonas syringae pv. maculicola, a plant pathogen. The IC strip detected 105 colony-forming units CFU/ml P. syringae pv. maculicola in pure culture within 10 min and had good specificity. Compared with sandwich enzyme-linked immunosorbent assay, the strip was equally sensitive; however, it is portable and time-efficient. On broccoli and radish seeds, the IC strips had a sensitivity of 5 × 105 CFU/ml and 106 CFU/ml, respectively. The results revealed that the IC strip could have potential applications for P. syringae pv. maculicola detection in plant seeds.
TL;DR: In this paper, a non-pretreatment monitoring method based on immunochromatographic test (ICT) strips was developed to test for chloramphenicol (CAP) in raw milk.
Abstract: A non-pretreatment monitoring method based on immunochromatographic test (ICT) strips was developed to test for chloramphenicol (CAP) in raw milk. This assay was designed to measure competitive binding affinities for CAP molecules in raw milk and CAP antigen immobilized on strips with colloidal gold nanoparticles (GNPs, average diameter of 30 nm) labeled with anti-CAP monoclonal antibody (CAP mAb). Several working conditions, including the CAP solvent, different types and concentrations of CAP mAb, coating antigen, and GNP, were optimized for performance. After optimization, the detection process was carried out in 8 min with a visual limit of detection (LOD) of 0.3 ng/mL for qualitative detection and a LOD of 0.064 ng/mL for semi-quantitative detection. No cross-reactivity was detected toward CAP analogs. Based on these results, this simple and ultrasensitive assay could be thoroughly applied in the on-site testing of CAP in raw milk.
TL;DR: The results show that the developed enzyme-linked immune technology based on 4D7 is a very useful analytical tool for the detection of chlorothalonil residues and is important to immunity studies on chlorothsalonil.
Abstract: A sensitive monoclonal antibody for chlorothalonil was created based on a new hapten, which was prepared by an innovative method described in this paper. The hapten was generated by the reaction of original chlorothalonil drugs with 6-aminocaproic acid, distinct from that using glycol and ethylenediamine reported in other papers. A cell strain, (No. 4D7), was obtained with IC50 0.36 ng/mL, which was the best sensitivity obtained to date. The good recovery obtained in tests for cucumber indicated that the method possesses excellent accuracy and sensitivity. These results show that the developed enzyme-linked immune technology based on 4D7 is a very useful analytical tool for the detection of chlorothalonil residues and is important to immunity studies on chlorothalonil.
TL;DR: The production of a specific monoclonal antibody against Nitrazepam (NZP) was synthesized, then the hapten was coupled with bovine serum albumin and ovalbumin by the diazotization method, and used as immunogen and coating antigen.
Abstract: We report the production of a specific monoclonal antibody against Nitrazepam (NZP). First, the hapten 7-aminonitrazepam (7-ANZP) was synthesized, then the hapten was coupled with bovine serum albumin and ovalbumin by the diazotization method, and used as immunogen and coating antigen. Factors affecting binding, ionic strength, pH, and methanol concentration in phosphate-buffered saline, were optimized. The monoclonal antibody had a satisfactory IC50 of 0.2 ng/mL for NZP. The cross-reactivities with other analogs were all lower than 0.1% except for 23% with 7-NZP, which suggested that the enzyme-linked immunosorbent assay method possessed high specificity. The recoveries were in the range of 84–95%, indicating that the method could be applied for the detection of NZP in urine.
TL;DR: The effects of epithelial metabolic activity, permeability, integrity, and the tight junction (TJ) expression induced by glycinin were conducted using porcine intestinal epithelial cells (IPEC-J2) model in this study and showed decreased metabolic activity and increased alkaline phosphatase activity.
Abstract: Humans and animals suffer a health threat to soybean allergy. The impaired mechanism of the intestinal barrier by food/feed allergen glycinin has not been clear. The effects of epithelial metabolic activity, permeability, integrity, and the tight junction (TJ) expression induced by glycinin were conducted using porcine intestinal epithelial cells (IPEC-J2) model in this study. The results showed the decreased metabolic activity (R2 = 0.964, p < 0.001) and the increased alkaline phosphatase activity (R2 = 0.528, p = 0.002) linearly correlated with glycinin levels (0–4 mg/mL). The trans-epithelial electrical resistance values declined stimulated by 4 mg/mL glycinin treatment in a time-dependent manner (0, 6, 12, or 24 h, p = 0.018), and also descended after 12 or 24 h glycinin treatment in dose-dependence (0, 0.5, 1, 2, or 4 mg/mL, p < 0.001). The glycinin-induced TJ expression of claudin-3, occludin, and ZO-1 were reduced (p < 0.05).
TL;DR: LFI is a novel and sensitive assay based on MG mAb that has applications for the rapid detection of total MG and CV in fish products and cross-reactivity with most MG analogs was negligible.
Abstract: We developed a sensitive lateral flow immunoassay (LFI) for the detection of total malachite green (MG) residues in fish tissues. LFI was based on a colloidal gold nanoparticle-labeled monoclonal antibody against MG , which was generated by fast immunization of carboxymalachite green-bovine serum albumin. The half-maximum inhibition concentration (IC50) of MG monoclonal antibody (MG mAb) was 0.057 ng/ml with a linear range of 0.020–0.162 ng/ml. Following the optimization of LFI, a qualitatively visual limit of detection of 1 ng/ml and a semi-quantitatively limit of detection of 0.418 ng/ml (IC50 of the calibration curve) were obtained for total MG residues in 8 min. Cross-reactivity with most MG analogs was negligible, expect with crystal violet (CV; 78.6% cross-reactivity). LFI is a novel and sensitive assay based on MG mAb that has applications for the rapid detection of total MG and CV in fish products.
TL;DR: The study showed that supplementing weaning piglets diets with AMP enhanced the humoral immune responses of weaned piglets by improving the levels of classical swine fever virus antibody, CH50, and immunoglobulins.
Abstract: The aim of this study was to investigate the effects of dietary antimicrobial peptide (AMP) supplementation on the serum IgG, IgM, IgA, classical swine fever antibody (CSF-Ab), and total complement (CH50) levels in weaned piglets. Ninety weaned piglets were randomly allotted to five groups for 28-day study. Blood samples were collected at 32, 39, 46, and 53 days of age. The results showed that AMP as dietary additive increased the levels of IgG, IgM, IgA, CSF-Ab, CH50 among the whole experimental period, and the effects of AMP supplementation were dose-dependent. The study showed that supplementing weaned piglets diets with AMP enhanced the humoral immune responses of weaned piglets by improving the levels of classical swine fever virus antibody, CH50, and immunoglobulins. We suggest the dosage of AMP was 500 mg/kg.
TL;DR: To produce broad-selective monoclonal antibody (Mab) for organophosphorus (OP) pesticides, heterologous enzyme-linked immunosorbent assays (ELISAs) were used for hybridoma screening and selectivity was determined by its molecular properties.
Abstract: To produce broad-selective monoclonal antibody (Mab) for organophosphorus (OP) pesticides, heterologous enzyme-linked immunosorbent assays (ELISAs) were used for hybridoma screening. Six Mabs with different cross-reactivity to OP pesticides were produced. Based on the broad-selective Mab 5F7, homologous and heterologous indirect competitive ELISAs were developed and the influence of hapten structure on assay selectivity was investigated. Moreover, the receptor–ligand interactions between 5F7 and OP pesticides were simulated to evaluate the broad-selectivity of the Mab. Results showed that, selectivity of the Mab was determined by its molecular properties, and the assay selectivity could be modified when the assay sensitivity was improved sufficiently by heterology. With the most suitable competitor, a broad-selective heterologous ELISA was developed. The IC50 values were estimated to be 7.06 ng/mL for parathion, 32.34 ng/mL for methyl parathion, 164.84 ng/mL for fenitrothion, 96.97 ng/mL for phoxim and 50...
TL;DR: To evaluate if there is a difference in the occurrence of asthma bronchiale, rhinitis, pollen allergy, food allergy and sensitization to basic food allergens, onset of atopic dermatitis (AD), family history about atopy, and duration of eczematic lesions in AD patients with the level of total immunoglobulin E (IgE) under or above 200 IU/ml, complete allergological and dermatological examination was performed.
Abstract: The objective of this study is to evaluate if there is a difference in the occurrence of asthma bronchiale, rhinitis, pollen allergy, food allergy and sensitization to basic food allergens, onset of atopic dermatitis (AD), family history about atopy, and duration of eczematic lesions in AD patients with the level of total immunoglobulin E (IgE) under or above 200 IU/ml. Complete allergological and dermatological examination was performed and the statistical evaluation of the relations among the patients with the level of IgE under 200 IU/ml and above 200 IU/ml was performed. Out of 277 patients suffering from AD, we included 87 men and 190 women; the average age was 25.9 years. From 277 patients, 92 patients (33%) have IgE under 200 IU/ml; in these patients, the occurrence of tested parameters is significantly lower in comparison to patients with IgE above 200 IU/ml.
TL;DR: The diagnostic work-up should comprise not only the laboratory methods, but also the diagnostic hypoallergenic diet and the challenge test.
Abstract: Few large studies concerning the importance of food allergy in adolescents and adult patients with atopic dermatitis exist. The evaluation of food allergy to egg white and yolk, peanuts, soy, cow milk and wheat in patients suffering from atopic dermatitis. Two hundred forty patients (70 men, 170 women) were examined. Complete dermatological and allergological examination was performed in all patients, including specific IgE, skin prick test and atopy patch test. The challenge test was performed according to the results of examinations with suspected foods. The food allergy to peanuts was confirmed in 20% of patients, to egg in 6%, to soy in 3.3%, to wheat in 2.5% and to milk in 0.8% – altogether in 65 patients (27.5%). The positive results in examinations without clinical symptoms of food allergy were recorded in another 78 patients (32.5%). The diagnostic work-up should comprise not only the laboratory methods, but also the diagnostic hypoallergenic diet and the challenge test.
TL;DR: In this paper, a highly sensitive and specific monoclonal antibody was prepared to detect semicarbazide (SEM), which was synthesized through condensation reactions of SEM and 4-carboxybenzaldehyde (4-CBA). The active ester method was employed to couple the hapten to bovine serum albumin, keyhole limpet hemocyanin and ovalbumin (OVA) to obtain conjugates of immunogens and coating antigens.
Abstract: A highly sensitive and specific monoclonal antibody was prepared to detect semicarbazide (SEM). Hapten 4-{[(aminocarbonyl)hydrazono]methyl}benzoic acid (4-CPSEM) was synthesised through the condensation reactions of SEM and 4-carboxybenzaldehyde (4-CBA). The active ester method was employed to couple the hapten to bovine serum albumin, keyhole limpet hemocyanin and ovalbumin (OVA) to obtain conjugates of immunogens and coating antigens. A novel hapten 2-[(aminocarbonyl)hydrazono]acetic acid (SEM-A) was also synthesised from SEM and oxoacetic acid to prepare a heterologous coating antigen (SEM-A-OVA). The linear ranges of inhibition curves in the optimised indirect competitive enzyme-linked immunosorbent assay (icELISA) method were in the range of 0.0071–0.056 ng mL−1 and 0.018–0.209 ng mL−1 for coating antigens SEM-A-OVA and 4-CPSEM-OVA, respectively. The IC50 was 0.019 ng mL−1 for SEM (SEM in the form of 4-nitrobenzaldehyde semicarbazone) and 0.13 ng mL−1 for 2-nitrobenzaldehyde semicarbazone, both of wh...
TL;DR: In this paper, an immunochromatographic lateral flow strip test using the competitive format was developed for the determination of chlorothalonil (CTN) residues in cucumber.
Abstract: An immunochromatographic lateral flow strip test using the competitive format was developed for the determination of chlorothalonil (CTN) residues in cucumber. The limit of detection was less than 100 ng/mL by visual assessment and was found to be 91.78 ± 0.17 ng/mL for quantitative detection using a test strip reader. The recoveries of test samples were from 89.26% to 102.02% and the coefficient of variation (%) was less than 4.75%. Parallel analysis of cucumber samples with CTN showed comparable results from the strip test and high performance liquid chromatography. The strip test requires 5–8 min to complete and provides a very useful screening method for the quantitative and qualitative detection of CTN residues in cucumber.
TL;DR: The developed FPIA method exhibited the potential for the rapid and accurate determination of carbofuran in agricultural and environmental samples.
Abstract: A simple and rapid method for determination of carbofuran was developed using the fluorescence polarisation immunoassay (FPIA). Three tracers with different lengths of bridge (0-, 2- and 6-carbon bridge) between the hapten molecule 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy)carbonyl]-amino]-butanoic acid (BFNB) and 5-aminofluorescein (AF), fluoresceinthiocarbamyl ethylenediamine (EDF), fluoresceinthiocarbamyl hexylenediamine (HDF), were synthesised and their binding response with anti-carbofuran-specific antibody were evaluated. The physicochemical parameters were optimised for the FPIA. The AF-labelled BFNB conjugate (BFNB-AF) was found to be the optimal tracer for FPIA of carbofuran. The detection limit of carbofuran, IC50 value and the working range were 2.3, 48.8 and 7.4−202.2 µg/L, respectively; and the reaction time was only 10 min. The average recovery from spiked water and vegetable samples was 86.9−95.4% and the mean coefficient of variation was 6.2% for inter-assay and 8.7% for intra-assay,...
TL;DR: It is demonstrated that kombucha tea had beneficial effects on EAE by lower incidence, attenuation in the severity, and also a delay in the onset of disease.
Abstract: Experimental autoimmune encephalomyelitis (EAE) is a mouse model for multiple sclerosis (MS), in which an inflammatory demyelination and axonal damage occurs. Kombucha tea is a fermented beverage made from kombucha mushroom, brewed tea, and sugar. In recent years kombucha tea has attracted interest due to its pharmacological properties like antioxidant effects. The aim of the present research was to test the therapeutic effect of kombucha tea in EAE. We induced EAE model in 18 female C57BL/6 mice by inoculation of myelin oligodendrocyte glycoprotein-35-55 (MOG35-55) in complete Freund’s adjuvant emulsion. Then, in order to ameliorate EAE symptoms, we used kombucha tea. During the course of study clinical evaluation was assessed, and on the day 21 post-immunization, for evaluation of nitric oxide (NO), total antioxidants capacity and tumor necrosis factor-alpha (TNF-α), blood samples were taken from the heart of mice. The mice were sacrificed and brains and cerebellums of mice were removed for histological...
TL;DR: It is demonstrated that N. sativa and TQ exert anti-proliferative and pro-apoptotic effects on ACHN cells in a concentration and time-dependent manner, which suggests their potential to be used as a new therapeutic strategy for renal cancers.
Abstract: Renal cell carcinoma (RCC) is the most common renal malignancy in adults and exhibits highly intrinsic and acquired resistance to standard therapeutic strategies. We sought to determine the anti-cancer activity of hydroalcoholic extract of Nigella sativa seeds (NSE) and thymoquinone (TQ). Human renal cell carcinoma (ACHN) and fibroblast L929 cell lines were treated with NSE and TQ, and cytotoxicity was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) assay. Cell death pattern was determined by annexin V and propidium iodine (PI)-staining methods. Exposure to NSE, TQ and cisplatin significantly inhibited the growth of ACHN cells and showed significant increase of early apoptotic cells. Normal cells were more resistant to NSE and TQ-induced effects. The present study demonstrates that N. sativa and TQ exert anti-proliferative and pro-apoptotic effects on ACHN cells in a concentration and time-dependent manner, which suggests their potential to be used as a new therapeutic strategy ...
TL;DR: Atopic dermatitis patients with confirmed food allergy suffer significantly more often from rhinitis, asthma bronchiale, persistent eczematic lesions and pollen allergy and have positive data about atopy in their family history.
Abstract: A few reports demonstrate the relationship between food allergy in atopic dermatitis patients and other allergic diseases and parameters. The objective of this study is to evaluate if some food allergens has the relationship to the occurrence of other atopic diseases and parameters. The following parameters were examined: food allergy to wheat flour, cow milk, egg, peanuts and soy; the occurrence of asthma bronchiale and rhinitis; duration of atopic dermatitis; family history; pollen allergy and onset of atopic dermatitis. The statistical evaluation of the relations among food allergy and monitored parameters was performed. Two hundred and seventy two patients were examined (87 men, 185 women). In general, atopic dermatitis patients with confirmed food allergy suffer significantly more often from rhinitis, asthma bronchiale, persistent eczematic lesions and pollen allergy and have positive data about atopy in their family history. Peanuts, soy and wheat are of great importance.
TL;DR: The IC50 value and other parameters obtained for this CAP McAb suggest that it is likely to have a promising future in further applications in combination with different analytical techniques.
Abstract: In this study, a highly sensitive monoclonal antibody (McAb) against chloramphenicol (CAP) was successfully raised and characterised by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). Three antigens coupled with different proteins were synthesised and determined by the UV-vis method. After five rounds of immunisation, antisera were collected from six Balb/C mice and screened by ic-ELISA. The standard inhibition curve of one CAP McAb (labelled 6F11) showed the best sensitivity to CAP after optimisation of the principal working conditions. The half-maximal inhibition concentration (IC50) was calculated to be 0.01 ng/mL and the linear working range was from 0.0028 ng/mL to 0.040 ng/mL with a correlation coefficient of 0.996. This McAb showed high specificity, with negligible cross-reactivities detected towards CAP analogues (florfinicol, florfinicol amine, thiamphenicol). The recovery rate was 98.9–106.8% while the coefficient of variation was 4.45–6.05%. The IC50 value and other parameter...
TL;DR: Mice exposed to leaf extracts from both MON810 and unmodified maize demonstrated influx of lymphocytes and eosinophils in the broncho-alveolar lavage, and increased cytokine release in mediastinal lymph node cells, indicating inherent immunogenicity and allergenicity.
Abstract: Genetically modified (GM) crops may bring new proteins with immunogenic and allergenic properties into the food and feed chains. The most commonly grown GM maize, MON810, expresses a modified version of the insecticidal Cry1Ab protein originating in the soil bacterium Bacillus thuringiensis (Bt). Immune reactions following inhalation of pollen and debris from such plants have been scarcely studied. We exposed BALB/c mice to purified Cry1Ab proteins and Cry1Ab-containing MON810 plant materials by intranasal installation. No anti-Cry1Ab antibodies were detected following exposure to the plant materials. Exposure to purified Cry1Ab resulted in specific anti-Cry1Ab IgG1 and IgE production, indicating inherent immunogenicity and allergenicity. Mice exposed to leaf extracts from both MON810 and unmodified maize demonstrated influx of lymphocytes and eosinophils in the broncho-alveolar lavage, and increased cytokine release in mediastinal lymph node cells. The results indicate that the airway exposure to Cry1Ab ...