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PubMed1986

A human auto-immune antibody specifically recognizing initiator methionine tRNA from yeast and higher eucaryotes.

Analysis of sera from 168 patients with autoimmune disorders revealed that one patient with Sjôgren's syndrome produced antibodies against deproteinized initiator methionine tRNA in addition to those against La protein. This anti-tRNAimet recognizes also tRNAimet from yeast but not from Phaseolus vulgaris chloroplasts (bean) or E. coli. It appears therefore that the epitope could be located in the TF loop in which an A residue in position 60 and the AUCG sequence are the only common features in yeast and human tRNAimet.

Autoantibodies↗
PubMed1992

Comoviruses and enteroviruses share a T cell epitope.

An in vitro murine T cell proliferation assay was used to determine whether an antigenic epitope(s) recognized by enterovirus-immune T cells is held in common between plant comoviruses and human enteroviruses. Splenocytes isolated from C3H/HeJ mice infected with coxsackievirus B3 (CVB3) proliferated in vitro not only against a variety of enterovirus (CVB2, CVB3, CVB6, CVA16, PV1) antigens, but against comovirus (CPMV, BPMV) antigens as well. Splenocytes from mice inoculated with bean pod mottle virus (BPMV) also proliferated in response to comoviral and enteroviral antigens in vitro. However, if the viral inocula were highly purified prior to inoculation, then the splenocyte response was generated only against the group used to inoculate, suggesting that the epitope shared between the comoviruses and the enteroviruses resided in the nonstructural region. B (nonstructural) and M (structural) genomic segments of CPMV were translated in rabbit reticulocyte lysates and used as in vitro antigens. Splenocytes from mice inoculated with live CVB3 proliferated in response to the B-RNA-encoded but not the M-RNA-encoded polypeptides, confirming the nonstructural coding region location of the common epitope. Comparison of predicted amino acid sequences in the nonstructural coding regions of the comoviruses and picornaviruses suggested a potentially immunogenic linear epitope in protein 2C. The consensus peptide LEEKGI was synthezized and shown to be immunogenic for both BPMV- and CVB3-immune splenocytes.

Amino Acid Sequence↗
PubMed1982

Mineral reserves in castor beans: the dry seed.

Elemental composition and distribution of the mineral reserves in the endosperm and embryo tissues of Ricinus communis cultivars Hale and Zanzibarensis were investigated. Energy dispersive x-ray analysis was used to determine the elemental composition of the globoid crystals, while atomic absorption spectrometry allowed quantification of the elements, particularly Ca, in various seed regions. No major differences were found between the two cultivars with regard to the elemental distribution in globoid crystals. While the majority of globoid crystals contained P, K, and Mg, the occasional one also contained Ca. In extremely rare instances, Fe was detected in globoid crystals. Ca-containing globoid crystals were more common in provascular cell protein bodies in the stem and radicle. Polarized light microscopy, micro-incineration, and acid solubility tests demonstrated the presence of calcium oxalate crystals in the innermost testa which adheres to the endosperm and is often mistakenly identified as endosperm. Atomic absorption spectrometry revealed that most of the calcium present in castor bean seeds is localized in the testa. On a perseed-region basis, the much larger endosperm contains more Ca than does the embryo. However, on a unit-weight basis, the radicle-plus-stem regions contain considerably more Ca than does the cotyledon or endosperm, an observation that is consistent with the observed distribution pattern for Ca-containing globoid crystals.

Journal Article↗
PubMed1971

Development of Microbodies in Sunflower Cotyledons and Castor Bean Endosperm during Germination.

In cotyledons of sunflower seedlings glyoxysomal and peroxisomal enzymes exhibit different rates of development during germination. The total activity of isocitrate lyase, a glyoxysomal marker enzyme, rapidly increased during the first 3 days, and then decreased 89% by day 9. Exposure to light accelerated this decrease only slightly. The specific activity of glyoxysomal enzymes (malate synthetase, isocitrate lyase, citrate synthetase, and aconitase) in the microbody fraction from sucrose density gradients increased between days 2 and 4 about 2- to 3-fold, and thereafter it remained about constant in light or darkness.Total activity of the peroxisomal enzymes increased slowly in the dark during the first 4 days of germination and thereafter remained at a constant level of activity in the dark or increased 2-fold in 24 hours of light. The specific activties of glycolate oxidase, hydroxypyruvate reductase, and serine-glyoxylate aminotransferase in the isolated microbody fraction increased about 10-fold between days 2 and 4 in the dark and then remained constant or increased again 10-fold after an additional 48 hours in the light.The total activity of the common microbody marker, catalase, developed similarly to isocitrate lyase, but decreased only 72% by day 9. The specific activities of enzymes (catalase, malate dehydrogenase, and aspartate aminotransferase) common to both microbody systems were 10- to 1000-fold greater than those of other enzymes. It is proposed that malate and aspartate may be involved in hydrogen transport between microbodies and other cellular sites.Glutamate-glyoxylate aminotransferase was very active in microbodies from castor bean endosperm and sunflower cotyledons. The specific activity of this aminotransferase developed similarly to glyoxysomal enzymes in the dark but further increased in the light, as did peroxisomal enzymes.The microbody fraction of castor bean endosperm germinated in the dark for 5 days contained both glyoxysomal and peroxisomal enzymes of similar specific activity.Adjacent to the microbody fraction on sucrose gradients from sunflower cotyledons were etioplasts at slightly lower densities and protein bodies at similar and higher densities. Their presence in the microbody fractions resulted in artificially low specific activities.

Journal Article↗
PubMed2002

Light strongly promotes gene transfer from Agrobacterium tumefaciens to plant cells.

Light conditions during Agrobacterium-based plant transformation, the most routinely used method in plant genetic engineering, differ widely and, to our knowledge, have not been studied systematically in relation to transformation efficiency. Here, light effects were examined in two already optimized transformation procedures: coculture of Agrobacterium tumefaciens with callus from two genotypes of the crop plant Phaseolus acutifolius (tepary bean) and coculture of root segments from two ecotypes of Arabidopsis thaliana. Except for the light conditions during coculture, all steps followed established procedures. Coculture was done either under continuous darkness, under a commonly used photoperiod of 16 h light/8 h darkness or under continuous light. beta-glucuronidase (GUS) production due to the transient expression of an intron-containing uidA gene in the binary vector was used to evaluate T-DNA transfer. In all situations, uidA expression correlated highly and positively with the light period used during coculture; it was inhibited severely by darkness and enhanced more under continuous light than under a 16 h light/8 h dark photoperiod. The promotive effect of light was observed with Agrobacterium strains harboring either a nopaline-, an octopine- or an agropine/succinamopine-type non-oncogenic helper Ti plasmid. The observed positive effect of light has obvious implications for developing and improving transient and stable transformation protocols, specifically those involving dark coculture conditions.

Agrobacterium tumefaciens↗
PubMed2000

Volatile aldehydes are promising broad-spectrum postharvest insecticides.

A variety of naturally occurring aldehydes common in plants have been evaluated for their insecticidal activity and for phytotoxicity to postharvest fruits, vegetables, and grains. Twenty-nine compounds were initially screened for their activity against aphids on fava bean leaf disks. Application under reduced pressure (partial vacuum) for the first quarter of fumigation increased insecticidal activity severalfold. The 11 best aldehydes were assayed against aphids placed under the third leaf of whole heads of iceberg lettuce using the same two-tier reduced-pressure regime, which caused no additional detriment to the commodity over fumigation at atmospheric pressure. Phytotoxicity to naked and wrapped iceburg lettuce, green and red table grapes, lemon, grapefruit, orange, broccoli, avocado, cabbage, pinto bean, and rice at doses that killed 100% of aphids was recorded for three promising fumigants: propanal, (E)-2-pentenal, and 2-methyl-(E)-2-butenal. These three compounds have excellent potential as affordable postharvest insect control agents, killing 100% of the aphids with little or no detectable harm to a majority of the commodities tested. Preliminary assays indicate that similar doses are also effective against mealybugs, thrips, and whitefly.

Aldehydes↗
PubMed1973

Nucleotide sequence analysis of the cytoplasmic 5S ribosomal ribonucleic acid from five species of flowering plants.

Broad-bean 5S rRNA labelled with (32)P was digested separately with T(1) and pancreatic A ribonucleases and the resulting oligonucleotides (20 and 18 respectively) were fractionated by two-dimensional electrophoresis. The oligonucleotides were analysed further and 32 of them have been completely sequenced. They were compared with those of 5S rRNA from dwarf bean, sunflower, tomato and rye. Sequence differences were found at both the 3'- and 5'-termini and at up to nine other positions. Most base substitutions were transitions between C and U. In common with other 5S rRNA species that have been analysed the ends of the molecule in each plant species have complementary sequences.

Autoradiography↗
PubMed1984

Mutagenic activity of nitrosated foods in an area with a high risk for stomach cancer.

In a search for possible sources of mutagens in human foods, a study was made of six common foods in the diet of a population at high risk for gastric cancer (in Nariño, Colombia). No mutagenic activity was demonstrated in nonnitrosated foods. After nitrosation, only one of the foods, fava beans, demonstrated a mutagenic effect. This effect was direct and concentrated in the germ and pulp of the beans.

Carcinogens↗
PubMedDate not supplied

Identification and characterization of the nolYZ genes of Bradyrhizobium japonicum.

Characterization of an isoflavone-inducible locus closely linked to the common nod genes of Bradyrhizobium japonicum USDA110 led to the discovery of two open reading frames, designated nolY and nolZ. These open reading frames are preceded by a sequence with strong similarity to a consensus NodD-binding site (nod box). Studies utilizing a nolZ'-'lacZ fusion indicated that inducible expression is dependent upon both NodD1 and NodW, transcriptional regulators that are required for the expression of the common nodulation genes (e.g., nodYABC) of B. japonicum. A deletion mutation within nolY produced only slight defects in nodulation of soybeans, siratro, and cowpeas, but stronger defects were observed in nodulation of mung beans. An insertion mutation within nolZ showed no nodulation defects in the host plants tested. Competition assays for nodule occupancy in soybeans did not show any decrease in the competitiveness of a nolY mutant, nor did a nolY mutant show any detectable alteration in the production of lipooligosaccharide nodulation signals.

Amino Acid Sequence↗
PubMedDate not supplied

Influence of pharmacogenetics on drug disposition and response.

1. Pharmacogenetics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare defects or as polymorphisms. 2. Glucose-6-phosphate dehydrogenase deficiency (G6PD) is the most common inherited enzymopathy affecting approximately 400 million people. The main clinical complication associated with G6PD deficiency is haemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans. 3. The activity of drug metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymorphisms of drug metabolizing enzymes (e.g. N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYP2C19) has provided an explanation as to why some patients do not obtain the expected drug effects or show exaggerated drug response and serious toxicity after taking the standard and safe dose of a drug. These polymorphisms are expressed in two phenotypes in the population, the extensive (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. The gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quite frequently experience exaggerated drug response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, PM show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. The other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.

Cytochrome P-450 Enzyme System↗
PubMed1992

Cell line specific abnormalities in expression of PNA, SBA and L-PHA binding sites by carcinogen induced rat urothelial carcinomas.

Bladder tumor cell lines derived from male F344 rats treated with N-buthyl N-(4-hydroxybuthyl) nitrosamine (BBN) or N-[4-(5-nitro-2-furyl)-2-thiazolyl] formamide (FANFT) have been established in vitro and characterized with respect to histology, karyotype, myc and c-Ha-ras oncogene expression or mutation, anchorage-independent growth and tumorigenicity in nude mice. This unique model system comprising 13 cell populations was employed to study common events during development of carcinogen-induced urothelial neoplasia. Differential expression of malignant phenotypes by these cell lines prompted us to examine their expression of carbohydrate structures binding peanut agglutinin (PNA), soy bean agglutinin (SBA) or leukoagglutinin (L-PHA), which are known indicators of tumor progression in rodents and humans. In the present study we analyzed the patterns of glycoproteins reactive with PNA and L-PHA by Western blotting. We also estimated quantitative differences in lectin binding to surfaces of normal rat urothelium and tumor cell lines by flow cytometry. The patterns of PNA or L-PHA reactive glycoproteins expressed by tumor cells were different from that of normal urothelium in culture. They were also different amongst the tumor cells. A unique non-sialylated, PNA binding glycoprotein (117 kD) was seen in the case of the highly tumorigenic F5 cell line and absent in normal urothelium as well as in other tumor cell lines. Normal cells did not express glycoprotein 60 kD binding PNA (only after desialylation), which was found in lysates of some but not all transformed cell lines. A very high molecular weight (much greater than 200), perhaps mucin-like sialoglycoprotein was found in normal urothelium but not in most of the tumor cell lines. Four major L-PHA reactive bands (greater than 200, 190, 100, 80 kD approximately) were found in normal urothelium. Some of those bands were overexpressed or missing in materials isolated from different tumor cell populations. Total cell surface binding of SBA and PNA by different tumor cell lines was very heterogenous (167-2% that of normal urothelium). No simple correlation between expression of the lectin binding glycoconjugates by urothelial carcinoma cells and other known functional, phenotypic or genetic alterations was found. We were also unable to demonstrate carcinogen-specific changes in expression of lectin binding to these tumor cell lines. Thus we conclude that lectin binding patterns are cell line specific. This may reflect distinct pathways of progression of individual cell lines. The potential sources of phenotypic variability between the cell lines were discussed.

Animals↗
PubMed2005

Food-specific IgG4 antibody-guided exclusion diet improves symptoms and rectal compliance in irritable bowel syndrome.

OBJECTIVE: Dietary modification improves symptoms in irritable bowel syndrome (IBS). Identification of offending foods by dietary elimination/re-challenge is cumbersome. IgG4 antibodies to common food antigens are elevated in IBS. The aim of this article was to evaluate the effect of exclusion diet based on IgG4 titres on IBS symptoms and rectal sensitivity and compliance. MATERIALS AND METHODS: The study comprised 25 patients with IBS (3 M, 22 F, mean age 43 years, Rome II criteria). IgG4 titres to 16 foods (milk, eggs, cheese, wheat, rice, potatoes, chicken, beef, pork, lamb, soya bean, fish, shrimps, yeast, tomatoes and peanuts) were measured. Foods with titres >250 microg/l were excluded for 6 months. Symptom severity was assessed with a previously validated questionnaire at baseline, at 3 months and at 6 months. Rectal compliance and sensitivity were measured in 12 patients at baseline and at 6 months. RESULTS: IgG4 antibodies to milk, eggs, wheat, beef, pork and lamb were commonly elevated. Significant improvement was reported in pain severity (p < 0.001), pain frequency (p = 0.034), bloating severity (p = 0.001), satisfaction with bowel habits (p = 0.004) and effect of IBS on life in general (p = 0.008) at 3 months. Symptom improvement was maintained at 6 months. Rectal compliance was significantly increased (p = 0.011) at 6 months but the thresholds for urge to defecate/discomfort were unchanged. CONCLUSIONS: Food-specific IgG4 antibody-guided exclusion diet improves symptoms in IBS and is associated with an improvement in rectal compliance.

Adult↗
PubMed2001

PFA, a novel mollusk agglutinin, is structurally related to the ribosome-inactivating protein superfamily.

The structural organization of PFA, a novel beta-galactose-specific agglutinin from the snail Pomacea flagellata, was partially characterized. Using mass spectrometry, the molecular weight of this glycoprotein was determined as 32,444 Da (7.4% carbohydrate). The agglutinin was found to form very large aggregates in solution, which were dissociated to monodisperse native-like dimers upon addition of polyethyleneglycol. The identity of the first 38 and the last 11 residues of the polypeptide chain was determined. It was found that PFA and the N-glycosidase subunit of ricin, a heterodimeric cytotoxin isolated from castor bean seeds, are homologous to each other in the N-terminal region. Furthermore, the far-UV circular dichroism spectra of these proteins were found to be nearly superimposable, evidencing that they share common general features in their secondary and tertiary structures. On the basis of these similarities, it can be concluded that PFA is structurally related to the ribosome-inactivating protein superfamily.

Agglutinins↗
PubMed1974

Immunological and biochemical studies on isozymes of malate dehydrogenase and citrate synthetase in castor bean glyoxysomes.

Rabbit gamma-globulin obtained after the injection of solubilized proteins of castor bean (Ricinus communis L var. Hale) glyoxysomes contains antibodies against some of the glyoxysomal enzymes. The gamma-globulin was shown to inhibit by 50% and 80%, respectively, the in vitro activities of the castor bean glyoxysomal citrate synthetase and malate dehydrogenase. The conditions required for the inactivation are described.The glyoxysomal and mitochondrial citrate synthetase from castor bean endosperm show no significant difference in the above immunological test, in their apparent Michaelis constant values for acetyl coenzyme A and oxaloacetate, or in their sensitivity toward ATP inhibition. In the immunological test, glyoxysomal malate dehydrogenase from castor bean endosperm appeared to be related more closely to the leaf peroxisomal malate dehydrogenase from spinach (Spinacia oleracea L.) and to the glyoxysomal malate dehydrogenase from a variety of other fatty seedlings than to the castor bean mitochondrial or soluble malate dehydrogenase. Unlike the mitochondrial or soluble malate dehydrogenase, the castor bean glyoxysomal malate dehydrogenase is easily inactivated by gentle heat treatment, as was reported previously by other workers for the leaf peroxisomal malate dehydrogenase. Thus, the malate dehydrogenases from various plant microbodies share some common characteristics which distinguish them from other isozymes of malate dehydrogenase.

Journal Article↗
PubMed2001

Reduced pulmonary function is associated with central arterial stiffness in men.

The association of impaired pulmonary function with cardiovascular morbidity and mortality has been reported in several prospective studies. The nature of this association and the mechanisms underlying it are unknown. Both atherosclerosis and central arterial stiffness might be involved. We recently reported, in a 4-yr longitudinal study, that reduced lung function predicts the development of carotid atherosclerotic plaques. In the present study, we report the associations of aortic stiffness with lung function measurements. One hundred and ninety-four men, aged 30 to 70 yr and free of coronary heart disease, who volunteered for a standard health examination were included. FEV(1) and FVC were used to assess lung function. Aortic stiffness was estimated from the carotid-femoral pulse-wave velocity (PWV), which increases proportionally with an increase in aortic stiffness. PWV was significantly and negatively associated with FEV(1) and FVC (partial correlation coefficients adjusted for age and height: -0.27 [p < 0.001] and -0.24 [p < 0.001], respectively). For every 1 SD increase in PWV (2.5 m/s), FEV(1) decreased by 195.2 +/- 50.1 ml (p < 0.001) in an age- and height-adjusted analysis. The corresponding decrease in FVC was 190.4 +/- 55.0 ml (p < 0.001). Further adjustment for cardiovascular risk factors (weight, smoking habits, hypercholesterolemia, diabetes, and hypertension) did not markedly alter these results. In addition, negative associations of PWV with lung function measurements were observed within each category of cardiovascular risk factors. This study suggests that reduced pulmonary function is independently associated with aortic stiffness in men. The interrelations between pulmonary and vascular alterations should be thoroughly investigated.

Adult↗
PubMed1980

Food habits and nutritional status of agricultural migrant workers in Southern Brazil.

A new class of migrant workers, commonly known as "Boia-Frias", is rapidly growing in the periurban slumbs (favelas) of Brazil. In 1978 a collaborative study was undertaken to assess the food habits and nutritional status of 100 migrant worker families of Vila Recreio, a typical Boia-Fria settlement near Ribeirao Preto in the state of Sao Paulo. The findings of this survey revealed that the traditional diet of Boia-Frias is nutritionally inadequate both in quality and quantity. Their rice and bean-based diet lacks sufficient variety because of the infrequent use of fresh fruits and vegetables, which are available locally, and of supplemental amounts of protein-rich foods of animal origin. Empty-calorie foods such as carbonated drinks and alcoholic beverages are consumed freely; and starchy foods, traditionally used in the North and Northeast of Brazil, are used commonly as weaning foods. Although dietary practices of pregnant and lactating women are poor, breast-feeding is still practiced by most mothers. The biochemical analysis of blood samples did not indicate major subclinical deficiencies except low hematological values and low plasma vitamin A concentrations in about 25% of the population examined. Plasma cholesterol and plasma vitamin E values were found to be normal. However, anthropometric examinations revealed clear signs of malnutrition and/or undernourishment, which likely impairs their capacity for physical work and adversely affects their overall health.

Adult↗
PubMed2002

The apoplastic oxidative burst in response to biotic stress in plants: a three-component system.

The oxidative burst, the generation of reactive oxygen species (ROS) in response to microbial pathogen attack, is a ubiquitous early part of the resistance mechanisms of plant cells. It has also become apparent from the study of a number of plant-pathogen interactions and those modelled by elicitor treatment of cultured cells that there may be more than one mechanism operating. However, one mechanism may be dominant in any given species. NADPH oxidases have been implicated in a number of systems and have been cloned and characterized. However, the enzyme system which is the major source of ROS in French bean (Phaseolus vulgaris) cells treated with a cell wall elicitor from Colletotrichum lindemuthianum, appears to be dependent on an exocellular peroxidase. The second component, the extracellular alkalinization, occurs as a result of the Ca(2+) and proton influxes and the K(+) efflux common to most elicitation systems as one of the earliest responses. The third component, the actual reductant/substrate, has remained elusive. The low molecular weight compound composition of apoplastic fluid was compared before and after elicitation. The substrate only becomes available some min after elicitation and can be extracted, so that by comparing the profiles by LC-MS it has been possible to identify possible substrates. The mechanism has proved to be complex and may involve a number of low molecular weight components. Stimulation of H(2)O(2) production was observed with saturated fatty acids such as palmitate and stearate without concomitant oxylipin production. This biochemical evidence is supported by immunolocalization studies on papillae forming at bacterial infection sites that show the peroxidase isoform present at sites of H(2)O(2) production revealed by cerium chloride staining together with the cross-linked wall proteins and callose and callose synthase. The peroxidase has been cloned and expressed in Pichia pastoris and has been shown to catalyse the oxidation reaction with the same kinetics as the purified enzyme. Furthermore, Arabidopsis plants transformed heterologously using the French bean peroxidase in antisense orientation have proved to be highly susceptible to bacterial and fungal pathogens. Thus it is possible that Arabidopsis is another species with the potential to mount an apoplastic oxidative burst and these transformed plant lines may be useful to identify the peroxidase that is responsible.

Arabidopsis↗
PubMed1996

Internal tremor in patients with Parkinson's disease.

Although sensory symptoms were not originally described in Parkinson's disease (PD), in recent years it has been increasingly recognized that painful sensations and paresthesias occur in approximately 40% of patients. It has been our observation that PD patients often describe a sensation of internal tremor, a feeling of tremor inside the chest, abdomen, arms, or legs that cannot be seen. We investigated the prevalence and characteristics of internal tremor by administering a questionnaire to 100 consecutive patients with PD and 50 age-matched controls seen in our movement disorders center. A sensation of internal tremor was present in 44% of this sample of PD patients and in 6% of the control population (p < 0.0001). The presence of internal tremor was unrelated to Unified Parkinson's Disease Rating Scale score, Hoehn and Yahr stage, duration of disease, or the presence of observable tremor. The frequency of other sensory symptoms (aching, tingling, burning) was higher in the PD patients with internal tremor (73%) than in those without (45%; p = 0.005). Internal tremor is associated with anxiety in 64% of patients (p < 0.0001). It was described as uncomfortable and was unrelieved by antiparkinsonian medication in three quarters of patients. A sensation of internal tremor is commonly reported by PD patients and should be recognized as a useful diagnostic factor in PD.

Aged↗