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PubMed1991

Comparison of the binding characteristics to BHK-21 cells of viruses representing the two Theiler's virus neurovirulence groups.

The binding characteristics of the highly virulent GDVII and less virulent BeAn strains of Theiler's murine encephalomyelitis viruses (TMEV) to whole BHK-21 cells were determined using a direct viral binding assay. The overall rates of association and dissociation of BeAn and GDVII viruses were similar. Using a saturation binding assay intended for multivalent ligands, such as picornaviruses, the number of binding sites per cell was calculated as 1.6 x 10(5). Competitive binding assays with both viruses showed one-way blocking. In addition, treatment of cell monolayers with neuraminidase reduced binding of BeAn virus by 90% but did not affect GDVII binding. Wheat germ agglutinin, a lectin which blocks binding to sialic acid and N-acetylglucosamine residues, substantially reduced binding of radiolabeled GDVII and BeAn viruses. Treatment of asialylated cells with O-glycanase further reduced the binding of BeAn virus, suggesting that O-linked oligosaccharides are involved in viral binding. These results suggest members of the two TMEV virulence groups share a common receptor but bind it differently.

Animals↗
PubMed2000

Tissue specificity of geminivirus infection is genetically determined.

The types of cells and tissues infected by a virus define its tissue tropism. Determinants of tissue tropism in animal-infecting viruses have been extensively investigated, but little is known about plant viruses in this regard. Some geminiviruses in the genus Begomovirus exhibit phloem limitation and are restricted to cells of the vascular system, whereas others can invade mesophyll tissue. To identify viral genetic determinants of tissue tropism, we established a model system using two begomoviruses and their common host plant, Nicotiana benthamiana. Analysis by DNA in situ hybridization confirmed that tomato golden mosaic virus invades mesophyll tissues in systemically infected leaves, whereas bean golden mosaic virus remains phloem limited. Through genetic complementation and analysis of recombinant hybrid viruses, we demonstrated that three genetic elements of tomato golden mosaic virus determine its mesophyll tissue tropism. A noncoding region of the viral genome is essential for the phenotype, but it must be accompanied by one of two different coding regions. To our knowledge, this is the first example documented in a plant virus of noncoding DNA sequences that determine tissue tropism.

DNA, Viral↗
PubMed2003

Pseudomonas syringae exchangeable effector loci: sequence diversity in representative pathovars and virulence function in P. syringae pv. syringae B728a.

Pseudomonas syringae is a plant pathogen whose pathogenicity and host specificity are thought to be determined by Hop/Avr effector proteins injected into plant cells by a type III secretion system. P. syringae pv. syringae B728a, which causes brown spot of bean, is a particularly well-studied strain. The type III secretion system in P. syringae is encoded by hrp (hypersensitive response and pathogenicity) and hrc (hrp conserved) genes, which are clustered in a pathogenicity island with a tripartite structure such that the hrp/hrc genes are flanked by a conserved effector locus and an exchangeable effector locus (EEL). The EELs of P. syringae pv. syringae B728a, P. syringae strain 61, and P. syringae pv. tomato DC3000 differ in size and effector gene composition; the EEL of P. syringae pv. syringae B728a is the largest and most complex. The three putative effector proteins encoded by the P. syringae pv. syringae B728a EEL--HopPsyC, HopPsyE, and HopPsyV--were demonstrated to be secreted in an Hrp-dependent manner in culture. Heterologous expression of hopPsyC, hopPsyE, and hopPsyV in P. syringae pv. tabaci induced the hypersensitive response in tobacco leaves, demonstrating avirulence activity in a nonhost plant. Deletion of the P. syringae pv. syringae B728a EEL strongly reduced virulence in host bean leaves. EELs from nine additional strains representing nine P. syringae pathovars were isolated and sequenced. Homologs of avrPphE (e.g., hopPsyE) and hopPsyA were particularly common. Comparative analyses of these effector genes and hrpK (which flanks the EEL) suggest that the EEL effector genes were acquired by horizontal transfer after the acquisition of the hrp/hrc gene cluster but before the divergence of modern pathovars and that some EELs underwent transpositions yielding effector exchanges or point mutations producing effector pseudogenes after their acquisition.

Bacterial Proteins↗
PubMed1987

Effects of anti-Ureaplasma urease antibody on homologous and heterologous urease activities.

Among organisms in the class Mollicutes only Ureaplasma species possess urease. Antiserum to urease of U. urealyticum strain T960 (CX8) was used to examine the cross-reactivity of urease from other Ureaplasma species, as well as urease of jack bean and several urease-possessing walled bacteria. Immunological cross reactivity was used to establish phylogenetic relationships between various antigens. The ability of monospecific anti-urease antibody to inhibit urease activity was examined. The antiserum inhibited urease activity of the homologous strain the least of any Ureaplasma tested. It is postulated that urease possesses a minimum of two sets of epitopes. Binding of antibody to one epitope causes inhibition of enzyme activity; this epitope is common to urease of all Ureaplasma species. Binding of antibody to the other epitope prevents binding to the inhibition epitope; this epitope is specific to U. urealyticum strain T960 (CX8). No inhibition was observed with urease from jack bean or several walled bacteria.

Animals↗
PubMed2001

M13 cloning of mung bean nuclease digested PCR fragments as a means of gap closure within A/T-rich, genome sequencing projects.

Obtaining the complete DNA sequence of a genome is often not straightforward. After standard shotgun sequencing strategies have been employed there are often gaps remaining and these can be the most intractable regions, frequently containing repeat sequences, "uncloneable" sequences and/or regions of potential secondary structure or differential base composition. In genomes with a high A/T content, such as Plasmodium falciparum and Dictyostelium discoideum, solving these gaps is a particularly difficult problem as the sequences concerned are "fragile" and easily denatured, commonly uncloneable and have a paucity of good oligonucleotide priming sites. Reported here is a simple, yet reliable method for determining the sequence of A/T-rich gap-spanning PCR products. This method relies on the slippage of the specificity of mung bean nuclease so that it digests A/T-rich double-stranded DNA into a set of deletion fragments that can then be cloned into M13, sequenced and the original sequence assembled therefrom.

AT Rich Sequence↗
PubMed1998

The relation of fasting insulin to blood pressure in a multiethnic population: the Miami Community Health Study.

PURPOSE: The aim of this study was to examine the associations among fasting insulin, adiposity, waist girth, and blood pressure among a nondiabetic multiethnic population. METHODS: A cross-sectional study was performed among 25-44-year-old African-Americans (n = 159), Cuban-Americans (n = 128), and non-Hispanic whites (n = 207) selected from Dade County, Florida. Fasting insulin levels were correlated with resting blood pressure level within each ethnic group. The separate effects of percentage body fat and waist girth on the association between blood pressure and insulin were analyzed in multiple linear regression and analysis of covariance. RESULTS: Fasting insulin was positively associated with systolic (r = 0.26-0.39; P < 0.01) and diastolic blood pressure (r = 0.19-0.30; P = 0.10 to P < 0.001) among women of all ethnic groups and among non-Hispanic white men (r = 0.27; P < 0.05). Stepwise linear regression analyses revealed statistically significant associations between systolic and diastolic blood pressure and fasting insulin level in non-Hispanic whites independent of other covariates, including sex and percentage body fat (P < 0.001). Fasting insulin was also independently and significantly related to systolic blood pressure among African-Americans (P = 0.02). Among Cuban-Americans, sex and percentage body fat were the main correlates of blood pressure level. Analysis of covariance revealed a relationship between insulin and blood pressure that was independent of waist girth among men and women. CONCLUSIONS: Fasting insulin level and blood pressure were positively associated among African-Americans and non-Hispanic whites. This association was not entirely due to the common association with percentage body fat or waist girth.

Adipose Tissue↗
PubMed2003

The source of ochratoxin A in Brazilian coffee and its formation in relation to processing methods.

A total of 408 Brazilian coffee samples was examined during the 1999 and 2000 coffee harvest seasons for the presence of ochratoxin A (OA) and fungi with the potential to produce it. Samples came from four regions: Alta Paulista (western area of São Paulo State), Sorocabana (southwest São Paulo State), Alta Mogiana (northeast São Paulo State) and Cerrado Mineiro (western area of Minas Gerais State). Cherries and beans were examined at different stages: immature, mature and overripe cherries from trees, overripe cherries from the ground and beans during drying and storage on the farm. For mycological studies, the cherries and beans were surface disinfected with chlorine, plated on Dichloran 18% Glycerol Agar at 25 degrees C for 5-7 days and analysed for the presence of Aspergillus ochraceus and closely related species, A. carbonarius and A. niger. More than 800 isolates of fungi belonging to these species were identified and studied for the ability to produce OA using the agar plug technique and thin layer chromatography (TLC). A. niger was the species found most commonly (63% of isolates of these three species), but only 3% of them produced OA. A. ochraceus also occurred commonly (31% of isolates), and 75% of those studied were capable of OA production, a much higher percentage than reported elsewhere. A. carbonarius was found (6% of isolates) only in Alta Paulista, the hottest region studied, and only from beans in the drying yard or in storage. However, 77% of the A. carbonarius isolates were capable of producing OA. Average infection rates for cherries taken from trees were very low, but were higher in fruit taken from the ground, from the drying yard and from storage, indicating infection by toxigenic species after harvest. The average OA content in 135 samples of mature cherries from trees, overripe from trees, overripe from the ground, drying yard and storage was 0.1, <0.2, 1.6, 2.1 and 3.3 microg/kg, respectively. Although individual OA levels varied widely, only 9 of the 135 samples analysed exceeded 5 microg/kg OA, with one sample of poor quality dried coffee in excess of 100 microg/kg OA. The causes of high contamination were investigated on the farms concerned and several critical points were found, relating both to local climatic conditions and the drying processes used.

Aspergillus↗
PubMed2004

Molecular identification of mutations in G6PD gene in patients with favism in Iran.

Glucose 6-phosphate dehydrogenase is a highly polymorphic enzyme encoded by a human X-linked gene (Xq2.8). This enzyme catalyses the first step of pentose phosphate pathway, that converts glucose 6-phosphate to 6-phosphogluconate with production of NADPH2. G6PD deficiency is the most common human metabolic inborn error affecting more than 400 million people world wide. The main clinical manifestations are acute hemolytic anemia and jaundice, triggered by infection or ingestion of Fava beans or oxidative drugs. A predominant variant of G6PD named Mediterranean is often associated with favism. This has been evident in several countries including Northern coastal provinces of Iran. Other current variants are Chatham and Cosenza. Molecular identification of the most prevalent mutations in G6PD gene was carried out in 71 males and females with G6PD deficiency. They were from Iranian Northern province of Golestan. DNA was extracted from blood samples and analyzed for known G6PD mutation by PCR and restriction fragment length polymorphisms (RFLP) technique. Adapting this method, revealed that Mediterranean mutation at nt 563(C-->T) is predominant in the area (69%) and 26.7% of patients have Chatham mutation at nt 1003(G-->A). Findings indicate a higher prevalence of these mutations, in Golestan compared to Mazandaran (66.2% Mediterranean and 19% Chatham mutation) and Gilan (86.4% Mediterranean and 9.71% Chatham mutations). Cosenza mutation at nt 1376(G-->C), by PCR-RFLP technique was not found among other 3 samples (4.3%). The similarity of these results with mutations in Italy indicates probable existence of a common ancestral origin in the observed populations.

Favism↗
PubMed2006

Sigmoid volvulus: diagnostic twists and turns.

OBJECTIVE: The purpose of this study was to describe the clinical profile of patients with sigmoid volvulus and highlight the diagnostic difficulties and pitfalls. METHODS: This was a retrospective descriptive study of patients with sigmoid volvulus seen at a community hospital over a 5-year period from July 1999 to July 2004. RESULTS: Twenty-eight cases of sigmoid volvulus were identified, of which 16 were male and 12 female. The median age was 74 years. Twenty-seven were Chinese and one was Malay. Seven out of 28 patients were from nursing institutes while the rest came from their own homes. Eleven were bedridden and chronic constipation was a problem for 17 patients. Co-existing medical illnesses, including neuropsychiatric conditions, were common. Abdominal distension and pain were the most frequent presentations (26 and 21 patients, respectively). Twelve patients had constipation and 11 had diarrhoea. Only nine had the complete typical triad of abdominal distension, pain and constipation. The classical coffee-bean sign was identified in nine out of 26 initial plain abdominal radiographs done at the emergency department and a further 11 were diagnosed after review by the surgical team in the ward. Another four were identified only after computed tomography scan of the abdomen and pelvis. The remaining two patients were diagnosed intra-operatively after they underwent emergency laparotomy. CONCLUSIONS: As our aged population continues to grow, sigmoid volvulus may be more commonly encountered. Emergency physicians should maintain a high index of suspicion and avoid the pitfalls so as to prevent delay in diagnosis and treatment.

Adult↗
PubMed1995

Elaboration of amylase activity and changes in paste viscosity of some common Indian legumes during germination.

The effect of germination upto 120 hours on malting loss, amylase activity and viscosity of nine common Indian legumes was investigated. The amylase activity increased on progressive germination in all legumes, the increase being particularly high for green gram, horse gram, moth bean and black gram. Malted samples had lower cooked paste viscosity than native ones. Samples with high amylase activity exhibited proportionately lower viscosity. Maximum reduction in viscosity was observed in green gram, followed by moth bean, horse gram, black gram and cowpea within 48 hours of germination. Malting losses ranged between 12 to 27 percent over a period of 48 hours in all legumes. Germination beyond 48 hours resulted in considerably higher malting losses without much effect on viscosity.

Amylases↗
PubMed1993

Impact of some commonly used Egyptian diets on plasma lipids profiles of rats.

Seven types of diets commonly consumed in Egypt were studied in hyperlipemic rats. Each of the seven diets were fed to a group of hyperlipemic rats. The diets contain white beans and whole wheat bread (Diet 1); cabbage and peas (2); spinach and carrots (3); whole wheat bread and bean sprouts (4); whole wheat bread, white beans and peas (5); white beans and carrots (6) and the last diet contains peas and carrots (7). The experiment continued for 5 weeks, at the end of which different plasma lipids and biological parameters were compared to hyperlipemic rats fed the control diet for 5 weeks. Results showed that plasma total cholesterol of rats of all groups showed significant decrease, except in rats fed diet 6 in which no significant change was noticed. Plasma total lipids of rats fed diet 5 decreased significantly. Plasma phospholipids of rats fed diets 3 and 7 increased significantly. Rats fed diet 3 showed decreased levels of plasma high density lipoprotein-cholesterol. Body weight gain of rats fed diets 1, 2 and 3 decreased significantly. Total food intake was noticed to decrease on feeding diets 1 and 6. Significant decrease was produced in food efficiency ratio of rats fed diets 1, 5 and 6. Feeding any type of the previous seven diets produced significant increase of feces weight.

Animals↗
PubMed2003

An enzyme electrode based on lipoxygenase immobilized in gelatin for selective determination of essential fatty acids.

An enzyme electrode for the specific determination of omega-3 and omega-6 fatty acids from the mixture of essential fatty acids (EFAs) was developed by using lipoxygenase (LOX) (EC 1.13.11.12) from soy beans in combination with a dissolved oxygen (DO) probe. The enzyme electrode showed different sensitivities for linoleic (LA) and alpha-linolenic acids (ALA), the most common essential fatty acids. Enzyme electrode response depends linearly on LA concentration between 12.8-160.5 microM and ALA concentration between 3.8-18.9 microM in borate buffer, 0.2 M at pH 9.0. However, in phosphate buffer 0.2 M at pH 6.0 linearity is in the range of 7.5-22.5 microM of ALA concentration at 5 minutes response times. Moreover, maximum electrode response was found in borate buffer at pH 9.0 and 30 degrees C.

Biosensing Techniques↗
PubMed1977

Electron-paramagnetic-resonance studies of leghaemoglobins from soya-bean and cowpea root nodules. Identification of nitrosyl-leghaemoglobin in crude leghaemoglobin preparations.

1. Leghaemoglobins from soya-bean (Glycine max) and cowpea (Vigna unguiculata) root nodules were purified by chromatography on DEAE-cellulose phosphate columns at pH8.0 and pH5.8, to avoid the relatively low pH (5.2) commonly used to purify these proteins. 2. E.p.r. (electron-paramagnetic-resonance) spectra of the fluoride, azide, hydroxide and cyanide complexes of these ferric leghaemoglobins were very similar to the spectra of the corresponding myoglobin derivatives, indicating that the immediate environment of the iron in leghaemoglobin and myoglobin is similar, an imidazole moiety of histidine being the proximal ligand to the haem iron [cf. Appleby, Blumberg, Peisach, Wittenberg & Wittenberg (1976) J. Biol. Chem.251, 6090-6096]. 3. E.p.r. spectra of the acid-metleghaemoglobins showed prominent high-spin features very near g=6 and g=2 and, unlike myoglobin, small low-spin absorptions near g=2.26, 2.72 and 3.14. The width of the g=6 absorption derivative at 10-20K was about 4-4.5mT, similar to the value for acid-methaemoglobin. In contrast, a recently published (Appleby et al., 1976) spectrum of acid-metleghaemoglobin a had less high-spin character and a much broader absorption derivative around g=6. 4. E.p.r. spectra of ferric leghaemoglobin nicotinate and imidazole complexes suggest that the low-spin absorption near g=3.14 can be attributed to a trace of ferric leghaemoglobin nicotinate, and those near g=2.26 and 2.72 are from an endogenous dihistidyl haemichrome. 5. A large e.p.r. signal at g=2 in all samples of crude leghaemoglobin was shown to be from nitrosyl-leghaemoglobin. A soya-bean sample contained 27+/-3% of the latter. A previously unidentified form of soya-bean ferrous leghaemoglobin a was shown to be its nitrosyl derivative. If this is not an artifact, and occurs in the root nodule, the nitrosyl radical may interfere with the function of leghaemoglobin.

Chromatography, DEAE-Cellulose↗
PubMed1980

Heterogeneity of broad bean legumin.

Legumins from various cultivars were fractionated on a DEAE-Sephadex A-50 column and their subunit structures were investigated. The results obtained indicated the heterogeneity of legumin molecular species. The nature of the heterogeneity was common to all cultivars examined from the standpoint of the molecular sizes of the subunits. Four groups of molecular species with molecular weights of 320 000, 350 000, 380 000 and 400 000 were detected by gradient gel electrophoresis. The smallest molecular species was composed of only three kinds of subunit, with molecular weights of 20 500, 23 000 and 36 000, and the largest one was composed of five kinds of subunit with molecular weights of 19 000, 23 000, 36 000, 49 000 and 51 000. All molecular species were composed of intermediary subunits which consisted of acidic and basic subunits. The intermediary subunits with molecular weights of 61 700, 59 800 and 48 000 are composed of the acidic subunits with molecular weights of 51 000, 49 000 and 36 000 and the basic subunits with molecular weights of 19 000, 19 000 and 23 000 or 20 500, respectively. The possible molecular species constructed from the subunit groups classified according to their sizes were presented.

Chemical Fractionation↗
PubMed1991

Inhibition of N- and L-type Ca2+ channels by the spider venom toxin omega-Aga-IIIA.

omega-Aga-IIIA, an 8.5-kDa peptide toxin isolated from the venom of Agelenopsis aperta, was found to be a highly potent inhibitor of Ca channels in cardiac muscle and in peripheral and central neurons of rats and frogs. Cardiac L-type Ca channels were completely (Kd approximately 0.6 nM) blocked by omega-Aga-IIIA. In sensory neurons, the toxin inhibited most high-threshold Ca current but not T-type Ca current. omega-Aga-IIIA blocked with similar potency (Kd approximately 1.5 nM) both omega-conotoxin GVIA-sensitive and dihydropyridine-sensitive current components but left a fraction (approximately 35%) of high-threshold current that was also resistant to omega-conotoxin and dihydropyridines. The toxin blocks N- and L-type channels with equal potency and therefore may identify a high-affinity binding site common to these two Ca channel types.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1999

Comparative study of lifestyles of residents in high and low risk areas for gastric cancer in Jiangsu Province, China; with special reference to allium vegetables.

There is a low risk area for gastric cancer in Jiangsu Province, China, where people frequently consume raw allium vegetables. As a first step epidemiological study to clarify the factors involved in the low incidence of gastric cancer, we conducted a comparative study of the ecological factors in a high risk area (HRA), Yangzhong, and a low risk area (LRA), Pizhou, using a questionnaire. Subjects were selected from the general population according to age and sex, and comprised 414 residents of the HRA and 425 residents of the LRA. Ecological factors were compared for the two areas by Cochran-Mantel-Haenszel method, age-adjusted. Smoking and drinking habits were found to be more common in the LRA. On the other hand, allium vegetables were consumed in the LRA much more frequently, with high consumption of raw vegetables fruit, tomatoes, kidney beans and soybean products. People who consumed garlic en masse 3 times or more per week were 82% of men and 75% of women in the LRA, and 1% of men and women in the HRA. The results of the survey suggest that frequent consumption of allium vegetables, in addition to other anticancer foods, may be a factor in low mortality for gastric cancer.

Adult↗
PubMedDate not supplied

[Legume cross-reactivity].

Legumes are dicotyledonous plants belonging to the Fabales order. The main distinctive characteristic of which is their fruit (legumen, seeds contained in pods). This botanical order is formed by three families: Mimosaceae, Caesalpiniaceae and Papilionaceae or Fabacea. The Papilionaceae family includes the most important allergenic species: Lens culinaris (lentil), Cicer arietinum (chick-pea), Pisum sátivum (pea), Arachis hipogea (peanut), Phaseolus vulgaris (bean) y Glycine max (soy). Legumes are an important ingredient in the Mediterranean diet. Among Spanish children, sensitivity to legumes is the fifth most prevalent food allergy. Lentil and chick-pea are the most frequent cause of allergic reactions to legumes in Spanish children. Legumes could be involved in severe allergic symptoms. The different legumes have structurally homologous proteins, but they are not all equally allergenic, thus making it difficult to distinguish in vitro and in vivo cross-reactivity. We have demonstrated by skin tests and CAP that most of the patients are sensitised to more than one species. We have demonstrated a great degree of cross-reactivity among lentil, chick-pea, pea and peanut by ELISA inhibition (> 50 % max. inhibition). Unlike the Anglo-Saxons population, this phenomenon implies clinical sensitisation for many Spanish children. The majority of our patients have had symptoms with more than one legume (median 3 legumes). Thirty-nine patients were challenged (open or simple blind) with two or more legumes and 32 (82 %) reacted to two or more legumes: 43,5 % to 3, 25,6 % to 2, 13 % to 4 legumes. Seventy three per cent of the patients challenged with lentil and pea had positive challenge to both, 69,4 % to lentil and chick-pea, 60 % to chick-pea and 64,3 % to lentil, chick-pea and pea simultaneously. Peanut allergy peanut can be associated to allergy to lentil, chick-pea and pea but less frequently. Contrarily, white bean and overall green bean and soy are well tolerated by children allergic to other legumes. In our study, 82 % of the children allergic to legumes had a sensitisation to pollen. Pea and bean are the legumes with more in vitro cross-reactivity with Lolium perenne, Olea europea and Betula alba. This cross-reactivity could be because of common antigenic determinants or due to the coexistence of pollen and legume allergy. Panallergens implication seems to be less probable. It is important to emphasize that in spite of an evident clinical and immunological cross-reactivity, the diagnosis of legume allergy should not be based only on specific IgE tests. The decision to eliminate one legume from the diet should be based on a positive oral food challenge.

Adolescent↗