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PubMed1992

Contact factor proteases and the complexes formed with alpha 2-macroglobulin can interfere in protein C assays by cleaving amidolytic substrates.

Plasma from women taking combined oral contraceptives and cold-activated plasma contain proteases which cleave chromogenic substrates in protein C assays in the absence of protein C activators such as Protac. This spontaneous activity makes a background substraction necessary and makes protein C (PC) assays less accurate. We investigated two commonly used substrates < Glu-Pro-Arg-pNA (S-2366) and 2AcOH.H-D-Lys(Cbo)-Pro-Arg-pNA (PC substrate) and found that cold-activated normal and protein C-deficient plasmas gave absorbance values up to 300 times higher than buffer blanks. FXIa cleaves these substrates but activity was not blocked by corn or lima bean trypsin inhibitors, soy bean trypsin inhibitor (SBTI), hirudin or epsilon-amino-n-caproic acid (EACA). Kaolin activation of normal, FXI, FIX, FVIII, FVII and protein C-deficient, but not of FXII or prekallikrein (PKK)-deficient plasmas led to cleavage of chromogenic substrate for protein C. The protein C substrates were cleaved by purified kallikrein and alpha- and beta-FXIIa. Immunoabsorption with alpha 2-macroglobulin (alpha 2M) antibodies removed 60% of the alpha 2M and 70% of the activity on PC Substrate. Gel filtration of normal plasma on Sephadex G-150 gave a single peak of protein C activity and antigen in the included volume. After cold activation of the fractions, a second protein C-like peak appeared in the void volume, but with no detectable protein C antigen. This peak coincided with alpha 2M (chromogenic and ELISA) and plasma kallikrein (S-2302), but FXII (measured with a substrate insensitive to kallikrein) eluted separately.(ABSTRACT TRUNCATED AT 250 WORDS)

Amides↗
PubMed1989

Characterization of Rhizobium phaseoli Sym plasmid regions involved in nodule morphogenesis and host-range specificity.

Two nodulation regions from the symbiotic plasmid (pSym) of Rhizobium phaseoli CE-3 were identified. The two regions were contained in overlapping cosmids pSM927 and pSM991. These cosmids, in a R. phaseoli pSym-cured strain background, induced ineffective nodules on Phaseolus vulgaris roots. Transconjugants of Rhizobium meliloti harbouring pSM991 induced nodule-like structures on bean roots, suggesting that this cosmid contains host-range determinants. Analysis of deletions and insertional mutations in the sequences of pSM991 indicated that the genes responsible for the induction and development of nodules in P. vulgaris are organized in two regions 20 kb apart. One region, located in a 6.8 kb EcoRI fragment, includes the common nodABC genes. The other region, located in a 3.5 kb EcoRI fragment, contains information required for host-range determination.

Blotting, Southern↗
PubMed1985

Legume agglutinins that bind to Rhizobium meliloti.

A protein found in seeds and roots of alfalfa (Medicago sativa) was implicated in the specificity of the infection process, based on its binding to the symbiont Rhizobium meliloti. We found an agglutinin with similar properties in seeds and roots of sweet clover (Melilotis alba). The sweet clover differed from alfalfa in nodulation by a mutant strain of R. meliloti, but the agglutinins were indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Rhizobium agglutination, and cross-reactivity to antibodies. Similar agglutinins binding R. meliloti were found in seeds of legumes from different cross-inoculation groups, including soybean (Glycine max), cowpea (Vigna unguiculata), pea (Pisum sativum L), and mung bean (Vigna mungo). The agglutinins from these legumes were recognized by antibodies raised against the agglutinins of alfalfa and sweet clover. Seeds of corn (Zea mays) and tomato (Lycopersicon esculentum) contained a protein similar to the legume agglutinin, but it did not react with the antibodies. We conclude that the alfalfa agglutinin is representative of a common legume protein and that there is no evidence for its role in specificity or nodule initiation.

Adsorption↗
PubMedDate not supplied

The impact of household preparations on the residues of pesticides in selected agricultural food commodities available in India.

Gas chromatographic multiresidue methods for simultaneous determination of organophosphorus, organochlorine, and organonitrogen pesticides were used to study the exposure of the Indian population to pesticide contamination at their actual dietary intakes. Selected agricultural commodities--5 kinds of vegetables (tomato, potato, okra, cabbage, and green beans), 6 kinds of cereals and pulses (rice, maize, wheat, red gram, black gram, and green gram), and 6 kinds of fruits (mango, orange, guava, banana, apple, and grapes)--readily available in Chennai City local markets--were studied for this purpose. Samples were fortified with known concentrations of various pesticides and subjected to household preparation methods commonly used in India. The impact of household preparation is very high, resulting in 65-95% decontamination of pesticides at different stages. Of 512 raw market samples analyzed, the organochlorine and organophosphorus pesticides present in 12 samples were removed during household preparations, resulting in residues well below the toxicologically acceptable limits.

Chromatography, Gas↗
PubMed1981

Phytohemagglutinin isolectin subunit composition.

The subunit compositions of individual phytohemagglutinin isolectins from red kidney bean Phaseolus vulgaris were examined by isoelectric focusing and sodium dodecyl sulfate electrophoresis on polyacrylamide gels. Isoelectric focusing reveals heterogeneous but unique and non-overlapping protein band patterns for each of the homotetrameric isolectins, E4 and L4. Isoelectric focusing of the intermediate isolectins which contain both subunits (E3L1, E2L2, and E1L3) show all the protein bands common to isolectins E4 or L4 in proportions relative to their suggested subunit compositions. Polyacrylamide gel electrophoresis in a continuous sodium dodecyl sulfate buffer system gives a single protein band for all of the isolectins. In contrast, a discontinuous sodium dodecyl sulfate buffer procedure resolves isolectins E4 and L4 into single major protein bands of apparent molecular weights 31 700 (+/-600) and 29 900 (+/-200), respectively. Each of the intermediate isolectins contained both protein bands and their relative proportion, as determined by absorbance scanning, confirms the phytohemagglutinin isolectin subunit compositions as E4, E3L1, E2L2, E1L3, and L4.

Electrophoresis, Polyacrylamide Gel↗
PubMed1995

Evidence for the location of a binding sequence for the alpha 2 beta 1 integrin of endothelial cells, in the beta 1 subunit of laminin.

To date no specific location on laminin 1 for the binding of alpha 2 beta 1 integrin has been described, although recent evidence supports a location in the E1XNd fragment of the cross region. We have identified a peptide sequence from this region, in the beta 1 chain of laminin 1, YGYYGDALR, which inhibits the adhesion of endothelial cells to laminin 1 and type-IV collagen. A structurally related sequence from the CNBr-cleaved fragment CB3 of the alpha 1 chain of collagen type IV, FYFDLR, inhibits endothelial cell adhesion to both collagen types I and IV and laminin 1. The CB3 fragment containing the FYFDLR sequence has been shown to contain binding sites for both alpha 1 beta 1 and alpha 2 beta 1 integrins. Present experiments with anti-integrin antibodies indicate that the alpha 2 beta 1 integrin on endothelial cells can account for all the cell binding to collagen types I and IV, and that this integrin makes a major contribution towards the adhesion of these cells to laminin 1. We therefore propose that the peptide FYFDLR participates in alpha 2 beta 1 binding to collagen type IV and that the putatively structurally similar peptide, YGYYGDALR, participates in alpha 2 beta 1 binding to laminin 1. This is the first account of structurally related peptide sequences from laminin 1 and type-IV collagen which show reciprocal inhibition of cell adhesion to either ligand and which might form part of a common integrin-binding site, as well as the first suggestion of a precise location contributing to the alpha 2 beta 1 integrin binding site on laminin 1.

Amino Acid Sequence↗
PubMed2002

Taenia solium metacestode viability in infected pork after preparation with salt pickling or cooking methods common in Yucatán, Mexico.

The cestode parasite Taenia solium is an important cause of foodborne infection throughout tropical and subtropical regions. Ingestion of pork meat infected with T. solium larvae can lead to taeniasis infection in humans. With tourism and the consumption of native food increasing, it is important to investigate potential risks of transmission associated with food preparation methods. In this study, traditional Mexican salt pickling and two methods of pork preparation (as roast pork [cochinita pibil] and in pork and beans [frijol con puerco]) were evaluated in order to determine their effects on T. solium cyst viability in infected tissue. In the control groups, all metacestodes isolated were 100% viable, and only small changes in pH (from 6.0 to 5.9) and temperature (29 to 30 degrees C) were recorded. No viable cysts were detected after 12 and 24 h of salt pickling. The pH of the meat during salting dropped from 6.0 to 5.3. Osmotic changes and dehydration from the salting, rather than a change in pH, could be considered the main cause of cyst death. Temperatures of >65 degrees C damaged T. solium metacestodes in roast pork and in pork and beans. The results of this study indicate that if traditional pork dishes are prepared properly, T. solium cysts are destroyed. The criteria used in this study to evaluate the viability of tissue cysts are discussed.

Animals↗
PubMed2001

Purification and characterization of isoforms of beta-galactosidases in mung bean seedlings.

Five isoforms of beta-galactosidase (EC 3.2.1.23), designated as beta-galactosidases I-V, were isolated from five-day-old mung bean (Vigna radiata) seedlings. Beta-galactosidases II and III were purified to electrophoretic homogeneity by a procedure involving acid precipitation, ammonium sulfate fractionation, chromatography on diethylaminoethyl-cellulose (DEAE-Cellulose) and con A-Sepharose. and chromatofocusing. Beta-galactosidases I, II and III have the same molecular mass of 87 kDa. comprising two nonidentical subunits with molecular masses of 38 and 48 kDa, while beta-galactosidases IV and V have molecular masses of 45 and 73 kDa, respectively. All the enzymes were active against p-nitrophenyl-beta-D-galactoside, and to a lesser extent, p-nitrophenyl-alpha-L-arabinoside and p-nitrophenyl-beta-D-fucoside. The enzymes were inhibited by D-galactono-1,4-lactone, D-galactose, Hg2+, Ag+ and sodium dodecyl sulfate (SDS). Beta-galactosidases I, II and III were shown to be competitively inhibited by either D-galactono-1, 4-lactone or D-galactose. Isoforms I, II and III have a common optimal pH of 3.6, while isoforms IV and V have pH optima at 3.8 and 4.0, respectively. Isoelectric points of isoforms I, II and III were 7.7, 7.5 and 7.3, respectively. Double immunodiffusion analysis indicated that beta-galactosidases I, II, III and V are immunologically similar to each other, while beta-galactosidase IV shares partially identical antigenic determinants with the other four isoforms. The purified beta-galactosidases II and III were capable of releasing D-galactose residue from the hemicellulose fraction isolated from mung bean seeds.

Chromatography, Affinity↗
PubMed1996

Partial characterization of the lysine rich 280kD globulin from common buckwheat (Fagopyrum esculentum Moench): its antigenic homology with seed proteins of some other crops.

A high molecular weight (HMW) globulin, the main storage protein in grains of the common buckwheat (Fagopyrum esculentum Moench) has been partially characterized. The protein exists as an oligomeric complex with molecular mass of 280 kD and is composed of three groups (alpha, beta and gamma of polypeptides with molecular weights ranging between 55-60, 32-44 and 16-29 kD respectively. The protein showed serological cross reactivity with globulins of soya bean, mung bean and pea. Antibodies raised against the purified protein did not recognize the storage proteins from broad bean, rice, wheat and grain amaranth. The 280 kD globulin of buckwheat grains belongs to the legumin family of proteins.

Chromatography↗
PubMed2003

Dietary patterns associated with risk factors for cardiovascular disease in healthy US adults.

BACKGROUND: Certain nutrients are well established as dietary risk factors for cardiovascular disease (CVD), but dietary patterns may be a better predictor of CVD risk. OBJECTIVE: This study tested the hypothesis that the complex dietary behaviors of US adults can be grouped into major dietary patterns that are related to risk factors for CVD. DESIGN: With the use of food-frequency questionnaire data from the third National Health and Nutrition Examination Survey, dietary patterns of healthy US adults (>/or =20 y old; n = 13 130) were identified by factor analysis. Log-transformed biomarker data were associated with major dietary patterns after control for confounding variables in regression analyses. All statistical analyses accounted for the survey design and sample weights. RESULTS: Of 6 dietary patterns identified, 2 patterns emerged as the most predominant: the Western pattern was characterized by high intakes of processed meats, eggs, red meats, and high-fat dairy products, and the American-healthy pattern was characterized by high intakes of green, leafy vegetables; salad dressings; tomatoes; other vegetables (eg, peppers, green beans, corn, and peas); cruciferous vegetables; and tea. The Western pattern was associated (P < 0.05) positively with serum C-peptide, serum insulin, and glycated hemoglobin and inversely with red blood cell folate concentrations after adjustment for confounding variables. The American-healthy pattern had no linear relation with any of the biomarkers examined. CONCLUSIONS: The identification of common dietary patterns among free-living persons is promising for characterizing high-risk groups at the US population level. The dietary patterns identified here are similar to those reported in other nonrepresentative samples and are associated with biomarkers of CVD risk, which confirms that dietary pattern analysis can be a valuable method for assessing dietary intakes when predicting CVD risk.

Adult↗
PubMed2004

Diagnostic errors by radiology residents in interpreting pediatric radiographs in an emergency setting.

BACKGROUND: There are few data regarding the frequency and type of diagnostic errors made by radiology residents and fellows ("trainees"). However, increasing interest in reducing medical errors highlights the need to analyze which areas of medical knowledge are most problematic for physicians-in-training, including radiology trainees. Once these areas are identified, they can be emphasized during training. OBJECTIVE: To quantify the diagnostic errors made by radiology trainees interpreting radiographs from a pediatric emergency department. MATERIALS AND METHODS: A total of 23,273 dictations of emergency radiographs performed over a 1-year period at a pediatric hospital were analyzed for corrections after staff interpretation and for type and incidence of missed abnormalities by radiology trainees. Errors were categorized by type of pathology and anatomic region. RESULTS: Of the 80 errors detected, 90% were false negatives and 69% were recurrent. Most errors (69%) involved the diagnosis of fractures and/or dislocations. Sixty-one percent of all recurrent errors involved buckle, Salter II, avulsion, and transverse fractures-yet these cases constituted only 3% of all cases seen during the study period. CONCLUSION: The most common errors made by radiology trainees can be identified. By targeting these errors, training programs can improve the quality and relevance of the education they provide.

Child↗
PubMed1992

Symptoms and risk factors of Helicobacter pylori infection in a cohort of epidemiologists.

To identify symptoms and risk factors associated with Helicobacter pylori infection, a cohort of 341 epidemiologists was studied. All subjects had one banked serum (collected between 1969 and 1987) and one recent serum sample (collected in 1988) evaluated for H. pylori immunoglobulin G by enzyme-linked immunosorbent assay; subjects provided information on gastrointestinal symptoms and risk factors for gastritis and peptic ulcer disease. Prevalence of infection decreased from the early 1970s to the present. Eleven subjects (3% of the total cohort) seroconverted during the interval between serum samples, giving a crude conversion rate of 0.49% per person-year (95% confidence interval, 0.3-0.9). Nonreactors on the 1988 serum sample described similar symptoms to reactors. However, subjects who seroconverted in the interval between the two serum samples were more likely than either persistent nonreactors [relative risk (RR), 4.1] or persistent reactors (RR, 3.7) to have experienced upper gastrointestinal symptoms in the interval years. Consumption of caffeinated beverages (RR, 4.6) and residence in the northeastern United States (RR, 5.3) seemed to increase risk for infection. Because pain was similarly common in H. pylori-positive and -negative patients, H. pylori cannot be summarily accepted as the cause of dyspeptic symptoms even when infection is confirmed.

Adult↗
PubMed1996

A single editing event is a prerequisite for efficient processing of potato mitochondrial phenylalanine tRNA.

In bean, potato, and Oenothera plants, the C encoded at position 4 (C4) in the mitochondrial tRNA Phe GAA gene is converted into a U in the mature tRNA. This nucleotide change corrects a mismatched C4-A69 base pair which appears when the gene sequence is folded into the cloverleaf structure. C-to-U conversions constitute the most common editing events occurring in plant mitochondrial mRNAs. While most of these conversions introduce changes in the amino acids specified by the mRNA and appear to be essential for the synthesis of functional proteins in plant mitochondria, the putative role of mitochondrial tRNA editing has not yet been defined. Since the edited form of the tRNA has the correct secondary and tertiary structures compared with the nonedited form, the two main processes which might be affected by a nucleotide conversion are aminoacylation and maturation. To test these possibilities, we determined the aminoacylation properties of unedited and edited potato mitochondrial tRNAPhe in vitro transcripts, as well as the processing efficiency of in vitro-synthesized potato mitochondrial tRNAPhe precursors. Reverse transcription-PCR amplification of natural precursors followed by cDNA sequencing was also used to investigate the influence of editing on processing. Our results show that C-to-U conversion at position 4 in the potato mitochondrial tRNA Phe GAA is not required for aminoacylation with phenylalanine but is likely to he essential for efficient processing of this tRNA.

Base Composition↗
PubMed1996

Effects of a program on symmetrical posture in patients with hemiplegia: A single-subject design.

OBJECTIVES: Asymmetrical posture during static stance has been identified as a common problem in persons with hemiplegia. This study examined the effect of an activity-based therapy regimen on symmetric weight bearing and midline position of center of gravity (COG) in three adult subjects with hemiplegia. METHOD: An ABAB single-subject design was used. The intervention program, including sanding in front of a standing table and play a bean bag game, was introduced for 30 min each day during each intervention phase. Quantitative measurements of the weight distribution and the midline position of COG were taken with the Balance Master System (version 2.20). RESULTS: Visual inspection and statistical analysis of the data revealed a significant improvement in symmetric weight distribution and midline position of COG. The study suggests that this program may be a promising alternative to a variety of postural rehabilitation programs for persons with hemiplegia. CONCLUSION: Insecurity caused by poor stabilization and abnormal reactions to body weight bearing in an antigravity position might contribute to asymmetric postures. Results of this study suggest that an activity-oriented program can be effective in helping the persons with hemiplegia achieve symmetric stances.

Aged↗
PubMed1989

Pattern of humoral reactivity to type II collagen in rheumatoid arthritis.

Humoral immunity directed against type II collagen (CII) is a common although not specific feature of rheumatoid arthritis (RA). We have shown that 10 to 15% of the sera either from RA patients (n = 88) or from healthy controls (n = 149) reacted with native human CII. Conversely, autoantibodies to the alpha-1 (II) chains were significantly more frequent in the RA group (26.1% versus 6.0%, P<0.001), suggestingthatdenaturedCII may bean autoantigenin RA. Thus, human CII was cleaved with cyanogen bromide (CB), and immunoblotting techniques were performed on 19 RA and 21 normal sera. Among the four major CB peptides, CB10 and CB11 were recognized by most of the sera tested without distinction between normal or RA sera. Inhibition experiments using an ELISA have shown that: (i) antibodies to the native CII molecule did not cross-react with those recognizing the CB peptides, and vice-versa; (ii) the binding of the sera to native CII was partially inhibited by pre-incubation with alpha-1 (II) chains, and vice-versa; (iii) pre-incubation of the sera with CB peptides partially blocked the binding to alpha-1 (II) chains, whereas pre-incubation of the sera with alpha-1 (II) chains totally inhibited the reactivity against CB peptides; and (iv) a substantial proportion of the epitopes recognized by anti-CII autoantibodies was neither species specific nor type specific. Taken together, these findings reveal the existence of several populations of anti-CII autoantibodies: some antibodies react exclusively with conformational determinants of the CII molecule, and others are directed towards linear structures of alpha-1 (II) chains.

Animals↗
PubMed1992

Effects of CNS stimulants on the in vivo release of the colocalized transmitters, dopamine and neurotensin, from rat prefrontal cortex.

The effect of CNS stimulant drugs on the in vivo release of the colocalized neurotransmitters dopamine and neurotensin in rat prefrontal cortex was studied using microdialysis. Amphetamine, methylphenidate and nomifensine all increased extracellular fluid (ECF) levels of dopamine; however, their effects of neurotensin varied. Amphetamine increased both ECF dopamine (514 +/- 82% of basal) and neurotensin (350 +/- 49% of basal); however, the neurotensin increase lagged behind the increase in dopamine suggesting a possible trans-synaptic effect. Methylphenidate increased both dopamine and neurotensin (226 +/- 26% and 151 +/- 14% of basal respectively) co-synchronously, suggesting exocytosis of vesicles containing both dopamine and neurotensin. The nomifensine-induced increase in dopamine (202 +/- 23% of basal) was similar to that of methylphenidate, whereas the increase in neurotensin was significantly delayed and of lower magnitude (134 +/- 20% of basal). These data suggest that dopamine and neurotensin in part share a common releaseable pool in the prefrontal cortex. Moreover, dopamine may act presynaptically to increase neurotensin release and the different behavioral profiles of these psychostimulants may in part relate to their different effects on neurotensin release.

Analysis of Variance↗
PubMed2004

alpha-Mannosidase-catalyzed synthesis of a (1-->2)-alpha-D-rhamnodisaccharide derivative.

Enzymatic transglycosylation using p-nitrophenyl alpha-D-rhamnopyranoside as the glycosyl donor and 6equiv of ethyl 1-thio-alpha-D-rhamnopyranoside as the glycosyl acceptor yielded a D-rhamnooligosaccharide derivative. The reaction was catalyzed by jack bean alpha-mannosidase in a 1:1 (v/v) mixture of 0.1 M sodium citrate buffer (pH4.5)-MeCN at 25 degrees C. The enzyme exhibited high catalytic activity for the reaction, to afford in 32.1% isolated yield (based on donor substrate) ethyl alpha-D-rhamnopyranosyl-(1-->2)-1-thio-alpha-D-rhamnopyranoside, which is a derivative of the common oligosaccharide unit of the antigenic lipopolysaccharides from Pseudomonas.

Chromatography, High Pressure Liquid↗
PubMed2002

Accumulation of HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine) in indigenous and agricultural plants grown in HMX-contaminated anti-tank firing-range soil.

To investigate their potential for phytoremediation, selected agricultural and indigenous terrestrial plants were examined fortheir capacity to accumulate and degrade the explosive octahydro-1 ,3,5,7-tetra nitro-1,3,5,7-tetrazocine (HMX). Plant tissue and soil extracts were analyzed for the presence of HMX and possible degradative metabolites using high-performance liquid chromatography with diode-array UV detection (HPLC-UV), micellar electrokinetic chromatography with diode-array UV detection (MEKC-UV), and HPLC with electrospray ionization mass spectrometry (LC-MS). The pattern of HMX accumulation for alfalfa (Medicago sativa), bush bean (Phaseolus vulgaris), canola (Brassica rapa), wheat (Triticum aestivum), and perennial ryegrass (Loliumperenne) grown in a controlled environment on contaminated soil from an anti-tank firing range was similar to that observed for plants (wild bergamot (Monarda fistulosa), western wheat grass (Agropyron smithii), brome grass (Bromus sitchensis), koeleria (Koeleria gracilis), goldenrod (Solidago sp.), blueberry (Vaccinium sp.), anemone (Anemone sp.), common thistle (Circium vulgare), wax-berry (Symphoricarpos albus), western sage (Artemisia gnaphalodes), and Drummond's milk vetch (Astragalus drummondii)) collected from the range. No direct evidence of plant-mediated HMX (bio)chemical transformation was provided by the available analytical methods. Traces of mononitroso-HMX were found in contaminated soil extracts and were also observed in leaf extracts. The dominant mechanism for HMX translocation and accumulation in foliar tissue was concluded to be aqueous transpirational flux and evaporation. The accumulation of HMX in the leaves of most of the selected species to levels significantly above soil concentration is relevant to the assessment of both phytoremediation potential and environmental risks.

Azocines↗