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PubMed1999

Genomic organization and expression properties of the MtSucS1 gene, which encodes a nodule-enhanced sucrose synthase in the model legume Medicago truncatula.

We have isolated and sequenced a sucrose synthase (SucS) cDNA from the model legume Medicago truncatula. This cDNA (MtSucS1) contains an ORF of 2418 bp, coding for a protein of 805 amino acids with a molecular mass of 92.29 kDa. The deduced amino acid sequence shows significant homology to other plant sucrose synthases, in particular to the nodule-enhanced sucrose synthases from pea and broad bean. Northern analysis revealed that the corresponding gene shows a ten-fold higher expression level in root nodules than in uninfected root, stem and leaf tissues. SucS protein was detected in root nodules from a variety of legumes, including M. truncatula. Whereas only one SucS isoform was detectable in root nodules, an additional sucrose synthase of slightly larger molecular weight was present in uninfected root, stem and flower tissues of M. truncatula. From our expression and sequence data we infer that the MtSucS1 gene encodes a nodule-enhanced sucrose synthase in M. truncatula. Southern hybridization data indicate that MtSucS1 is a single-copy gene. An analysis of a genomic MtSucS1 sequence revealed that the gene consists of 14 exons with the start codon being located on exon II. As is common for SucS genes, the MtSucS1 gene contains a large intron of 747 bp in the 5' untranslated region. The transcriptional start of MtSucS1 was mapped and putative regulatory elements in the MtSucS1 promoter were identified.

Amino Acid Sequence↗
PubMed1990

Bean pathogenesis-related (PR) proteins deduced from elicitor-induced transcripts are members of a ubiquitous new class of conserved PR proteins including pollen allergens.

We have searched for induced transcripts in a cDNA library derived from bean cell supension cultures treated with an elicitor from Colletotrichum lindemuthianum. Six independently isolated cDNAs corresponding to rapidly induced small mRNAs have been classified by their DNA sequence and slightly different induction behaviour into two groups. 5'- and 3'-untranslated regions exhibit little similarity, but the deduced small acidic proteins designated PvPR1 and PvPR2 are 89% identical. No relationship was found with the well-characterized PR1 proteins from tobacco. However, the PvPR proteins are closely related to pI49 in pea (64% identity), pSTH2 in potato (41% identity) and PcPR1-1 in parsley (39% identity), which are also induced in response to elicitor or microbial attack. Moreover, a major pollen allergen in birch (BetvI) has a 44% identity with PvPR1 proteins. These similarities establish a ubiquitous class of conserved defense-related proteins and suggest a common yet still unknown function. Southern blot analysis indicates that PvPR protein gene organization is highly complex with an estimated copy number of more than 12 genes.

Amino Acid Sequence↗
PubMed1982

Delayed hypersensitivity testing for the prediction of postoperative complications.

One hundred and sixty-six patients undergoing elective major laparotomy were skin tested preoperatively with four common recall antigens in an attempt to correlate preoperative cell-mediated immune status with postoperative septic complications. Nineteen patients were anergic, 22 relatively anergic and the remaining 125 reacted to two or more of the antigens and were regarded as normally reactive. No significant differences in morbidity or mortality were found between patients who had depressed delayed cutaneous hypersensitivity reactions and those who reacted normally. We conclude that identification of those patients with depressed cell-mediated immunity preoperatively does not help in predicting postoperative problems.

Adult↗
PubMed1986

Vitamin A status of young children in Southern Brazil.

A comprehensive survey was carried out to assess the vitamin A status of preschool children of poor migrant families in the periurban population of Ribeirao Preto, a typical agricultural town in the sugarcane and coffee region of the State of Sao Paulo in Southern Brazil. The intake of vitamin A and carotenoids from the rice and bean based diet of these children is considered low and appears to influence blood concentrations and liver reserves of this vitamin. With respect to plasma vitamin A, 1.8% of the children had a deficient level (less than 10 micrograms%), whereas 48.8% of the children had a low level (less than 20 micrograms%). Most children with inadequate plasma vitamin A (less than 20 micrograms%) responded positively to a massive dose of 200,000 IU vitamin A, suggesting that these children may be at risk of having low liver stores of vitamin A. Rose Bengal staining test and rapid dark adaptation time did not indicate definite signs of conjunctival xerosis and night blindness among these children. No ocular evidence of hypovitaminosis A was found in the children studied, but marginal or inadequate vitamin A status appears to be a common public health problem among young children in this region of Brazil.

Anthropometry↗
PubMed1972

Food iron absorption measured by an extrinsic tag.

The paper describes the use of an extrinsic tag of inorganic radioiron to determine the total absorption of nonheme iron from a complete meal. The method was developed by measuring the iron absorbed from vegetable foods containing biosynthetically incorporated (55)Fe (intrinsic tag) and from (59)Fe added as a small dose of inorganic iron to the same meal (extrinsic tag). In studies with maize, black bean, and wheat, a consistent extrinsic: intrinsic radioiron absorption ratio averaging 1.10 was observed. Similar results were obtained with either ferrous or ferric iron as the extrinsic tag, and with doses of the latter ranging from 0.001 to 0.5 mg iron added to a test meal containing 2-4 mg of food iron. Adding the radioiron at different stages in preparation of the test meal also had little effect. Separate administration of the extrinsic tag was less satisfactory when small portions of a single food were employed, but with a complete meal, the separate dose was preferable. The extrinsic tag provided a valid measure of absorption despite marked differences in the iron status of the subject, and with wide changes in absorption imposed by adding desferrioxamine or ascorbic acid to the test meal. These findings indicate that there is a common pool of nonheme iron, the absorption of which is influenced by various blocking or enhancing substances present in the meal.

Adult↗
PubMed2002

Foodborne disease outbreaks in United States schools.

BACKGROUND: The objective of this study was to describe the epidemiology of foodborne disease outbreaks in schools and to identify where preventive measures could be targeted. METHODS: Reports by state and local health departments of foodborne disease outbreaks occurring in primary and secondary schools, colleges and universities from January 1, 1973, through December 31, 1997, were reviewed. Data from ill persons identified through foodborne outbreak investigations and subsequently reported to the Centers for Disease Control and Prevention in the Foodborne Outbreak Surveillance System were examined. The number and size of foodborne disease outbreaks, as well as the etiologic agents, food vehicles of transmission, site of food preparation and contributing factors associated with outbreaks were also examined. RESULTS: From 1973 through 1997, states and local health departments reported 604 outbreaks of foodborne disease in schools. The median number of school outbreaks annually was 25 (range, 9 to 44). In 60% of the outbreaks an etiology was not determined, and in 45% a specific food vehicle of transmission was not determined. Salmonella was the most commonly identified pathogen, accounting for 36% of outbreak reports with a known etiology. Specific food vehicles of transmission were epidemiologically identified in 333 (55%) of the 604 outbreaks. The most commonly implicated vehicles were foods containing poultry (18.6%), salads (6.0%), Mexican-style food (6.0%), beef (5.7%) and dairy products excluding ice cream (5.0%). The most commonly reported food preparation practices that contributed to these school-related outbreaks were improper food storage and holding temperatures and food contaminated by a food handler. CONCLUSIONS: Strengthening food safety measures in schools would better protect students and school staff from outbreaks of foodborne illness. Infection control policies, such as training and certification of food handlers in the proper storage and cooking of foods, meticulous hand washing and paid sick leave for food handlers with gastroenteritis, could make meals safer for American students.

Adolescent↗
PubMed1989

A plant fucosyltransferase with human Lewis blood-group specificity.

A fucosyltransferase from mung bean seedling was found to transfer L-fucose from GDP-fucose to the Type 1 disaccharide beta-D-Galp-(1----3)-beta-D-GlcpNAc-OR [R = (CH2)8COOMe]. The product, which was detected by an anti-Lea antibody in a novel ELISA assay, was isolated and shown to be the human Lea blood-group determinant beta-D-Galp-(1----3)-[alpha-L-Fucp-(1----4)]-beta-D-GlcpNAc-OR by 1H-n.m.r. spectroscopy. This enzyme activity is distinct from that of the human Lewis-fucosyl-transferase since alpha-L-Fucp-(1----2)-beta-D-Galp-(1----4)-beta-D-Glcp-OR is a very poor substrate, while the Type 2 disaccharide beta-D-Galp-(1----4)-beta-D-GlcpNAc-OR is not an acceptor. In common with the Lewis fucosyltransferase, the H-Type 1 trisaccharide alpha-L-Fucp-(1----2)-beta-D-Galp-(1----3)-beta-D-GlcpNAc-OR was an excellent substrate for the enzyme. This new enzyme activity was further characterized with respect to pH, nucleotide, Mn2+ dependence, and acceptor specificity against a panel of synthetic oligosaccharides.

Carbohydrate Sequence↗
PubMed2003

Women's sexual health: an interdisciplinary approach to treating low sexual desire.

Hypoactive sexual desire is one of the most common sexual problems in women. This article describes an interdisciplinary, biopsychosocial model for assessing and treating hypoactive sexual desire in women. Developed at the University of Minnesota's Center for Sexual Health, the collaborative model can be implemented by physicians and other medical practitioners. This article focuses on the medical and psychopharmacological aspects of the approach and includes a brief description of the psychological treatment components. A key aspect of the approach is frequent communication and coordination of care among the team members, which enables all team members to reinforce all aspects of the treatment.

Combined Modality Therapy↗
PubMed1994

Pericoronal radiolucencies with and without radiopacities.

Pericoronal radiolucencies are common entities of the jaws that account for about 15% to 20% of all biopsy specimens. They are usually slow-growing, benign entities that involve odontogenic epithelium and the crown of a nonerupting tooth. Many lesions develop early in life and are detected during routine radiographic examination. Others are found in later years when a patient presents with a slow-growing swelling. The dentist is often the first provider to encounter these conditions. Thus, he or she must be familiar with their appearance and understand the proper management required.

Dental Sac↗
PubMed1991

Silencer region of a chalcone synthase promoter contains multiple binding sites for a factor, SBF-1, closely related to GT-1.

Bean nuclear extracts were used in gel retardation assays and DNase I footprinting experiments to identify a protein factor, designated SBF-1, that specifically interacts with regulatory sequences in the promoter of the bean defense gene CHS15, which encodes the flavonoid biosynthetic enzyme chalcone synthase. SBF-1 binds to three short sequences designated boxes 1, 2 and 3 in the region -326 to - 173. This cis-element, which is involved in organ-specific expression in plant development, functions as a transcriptional silencer in electroporated protoplasts derived from undifferentiated suspension-cultured soybean cells. The silencer element activates in trans a co-electroporated CHS15-chloramphenicol acetyl-transferase gene fusion, indicating that the factor acts as a repressor in these cells. SBF-1 binding in vitro is rapid, reversible and sensitive to prior heat or protease treatment. Competitive binding assays show that boxes 1, 2 and 3 interact cooperatively, but that each box can bind the factor independently, with box 3 showing the strongest binding and box 2 the weakest binding. GGTTAA(A/T)(A/T)(A/T), which forms a consensus sequence common to all three boxes, resembles the binding site for the GT-1 factor in light-responsive elements of the pea rbcS-3A gene, which encodes the small subunit of ribulose bisphosphate carboxylase. Binding to the CHS15 -326 to -173 element, and to boxes 1, 2 or 3 individually, is competed by the GT-1 binding sequence of rbcS-3A, but not by a functionally inactive form, and likewise the CHS sequences can compete with authentic GT-1 sites from the rbcS-3A promoter for binding.(ABSTRACT TRUNCATED AT 250 WORDS)

Acyltransferases↗
PubMed1996

Increased pathogenicity in a pseudorecombinant bipartite geminivirus correlates with intermolecular recombination.

Most whitefly-transmitted geminiviruses possess bipartite DNA genomes, and this feature may facilitate viral evolution through pseudorecombination and/or recombination. To test this hypothesis, the DNA-A and DNA-B components of the geminiviruses bean dwarf mosaic virus (BDMV) and tomato mottle virus (ToMoV) were exchanged, and the resultant pseudorecombinants were serially passaged through plants. Both pseudorecombinants were infectious in Nicotiana benthamiana but induced attenuated symptoms and had reduced DNA-B levels. Serial passage experiments revealed that the BDMV DNA-A plus ToMoV DNA-B pseudorecombinant could not be maintained beyond three passages. In contrast, the ToMoV DNA-A plus BDMV DNA-B pseudorecombinant was maintained during serial passage through N. benthamiana and Phaseolus vulgaris and, after three to five passages, became highly pathogenic. Furthermore, the increased pathogenicity of this pseudorecombinant was consistently associated with an increased level of DNA-B, which eventuated in equivalent levels of both components. Sequence analysis of the DNA-B component of the more pathogenic pseudorecombinant revealed that intermolecular recombination had taken place in which most of the BDMV DNA-B common region was replaced with the ToMoV DNA-A common region. This recombinant DNA-B component, which contained the ToMoV origin of replication, was the predominant DNA-B component associated with the more pathogenic pseudorecombinant. These results provide the first demonstration of recombination between distinct bipartite geminiviruses and establish that the bipartite genome can facilitate viral evolution through pseudorecombination and intermolecular recombination.

Base Sequence↗
PubMed2000

Structural analysis of free N-glycans occurring in soybean seedlings and dry seeds.

The structures of unconjugated or free N-glycans in stems of soybean seedlings and dry seeds have been identified. The free N-glycans were extracted from the stems of seedlings or defatted dry seeds. After desalting by two kinds of ion-exchange chromatography and a gel filtration, the free N-glycans were coupled with 2-aminopyridine. The resulting fluorescence-labeled (PA-) N-glycans were purified by gel filtration, Con A affinity chromatography, reverse-phase HPLC, and size-fractionation HPLC. The structures of the PA-sugar chains purified were analyzed by the combination of two-dimensional sugar chain mapping, jack bean alpha-mannosidase digestion, alpha-1,2-mannosidase digestions, partial acetolysis, and ESI-MS/MS. The free N-glycan structures found showed that two categories of free N-glycans occur in the stems of soybean seedlings. One is a high-mannose type structure having one GlcNAc residue at the reducing end (Man 9 approximately 5 GlcNAc1, 93%), that would be derived by endo-GM (Kimura, Y. et al., Biochim. Biophys. Acta, 1381, 27-36 (1998)). The other small component is a xylose-containing type one having two GlcNAc residues at the reducing end (Man3Xyl1GlcNAc2, 7%), which would be derived by PNGase-GM (Kimura, Y. and Ohno, A., Biosci. Biotechnol. Biochem., 62, 412-418 (1998)). The detailed structural analysis of free glycans showed that high-mannose type free N-glycans (Man 9 approximately 5 GlcNAc1) in the soybean seedlings have a common core structural unit; Manalpha1-6(Man1-3)Manalpha1-6(Manalpha1-3)Ma nbeta1-4GlcNAc. Comparing the amount of free N-glycans in the seedling stems and dry seeds, the amount in the stems of seedlings was much higher than that in the dry seeds; approximately 700 pmol per one stem, 8 pmol in one dry seed. This fact suggested that free N-glycans in soybean seedlings could be produced by two kinds of N-glycan releasing enzymes during germination or seedling-development.

Acetylglucosamine↗
PubMed1967

Fine structure of protein-storing plastids in bean root tips.

The fine structure of leucoplasts in root tip cells of Phaseolus vulgaris L. has been studied in material fixed in glutaraldehyde followed by osmium tetroxide and poststained in uranyl acetate and lead citrate. Plastid development has been followed from the young stages in and near the meristematic region, through an ameboid stage, to the larger forms with more abundant storage products in the outermost cells. The plastids contain a dense stroma penetrated by tubules and cisternae arising from the inner membrane of the plastid envelope. Also located in the stroma are lamellae, ribosome-like particles, phytoferritin granules, and fine fibrils in less dense regions. In some elongate plastids microfilaments run lengthwise in the stroma near the surface. The same plastids store both starch and protein, but in a strikingly different manner. The starch is deposited in the stroma, while the protein always is accumulated within membrane-bounded sacs. These sacs arise as outgrowths from a complex of interconnected tubules which in turn appears to originate by coalescence and proliferation of tubules and cisternae arising from the inner plastid membrane. This "tubular complex" bears a strong resemblance to the prolamellar body of etiolated chloroplasts, but is smaller and ordinarily less regularly organized, and is apparently light-insensitive. Crystallization of the protein commonly occurs in the sacs and occasionally takes place within the tubules of the complex as well. The fine structure of the leucoplasts is discussed in relation to that of etiolated chloroplasts. Suggestions are made concerning the function of the tubular complex, role of the ameboid plastid forms, and manner of accumulation of the storage protein in the plastids.

Cell Differentiation↗
PubMed2006

Translational control genes in the sea urchin genome.

Sea urchin eggs and early cleavage stage embryos provide an example of regulated gene expression at the level of translation. The availability of the sea urchin genome offers the opportunity to investigate the "translational control" toolkit of this model system. The annotation of the genome reveals that most of the factors implicated in translational control are encoded by nonredundant genes in echinoderm, an advantage for future functional studies. In this paper, we focus on translation factors that have been shown or suggested to play crucial role in cell cycle and development of sea urchin embryos. Addressing the cap-binding translational control, three closely related eIF4E genes (class I, II, III) are present, whereas its repressor 4E-BP and its activator eIF4G are both encoded by one gene. Analysis of the class III eIF4E proteins in various phyla shows an echinoderm-specific amino acid substitution. Furthermore, an interaction site between eIF4G and poly(A)-binding protein is uncovered in the sea urchin eIF4G proteins and is conserved in metazoan evolution. In silico screening of the sea urchin genome has uncovered potential new regulators of eIF4E sharing the common eIF4E recognition motif. Taking together, these data provide new insights regarding the strong requirement of cap-dependent translation following fertilization. The genome analysis gives insights on the complexity of eEF1B structure and motifs of functional relevance, involved in the translational control of gene expression at the level of elongation. Finally, because deregulation of translation process can lead to diseases and tumor formation in humans, the sea urchin orthologs of human genes implicated in human diseases and signaling pathways regulating translation were also discussed.

Amino Acid Sequence↗
PubMed2002

Biliary tract injury in laparoscopic cholecystectomy: results of a single unit.

BACKGROUND: Laparoscopic cholecystectomy was introduced into Australia in early 1990. Its rapid increase in acceptance was, however, tempered by reports of an increased incidence of bile duct injury. The aim of this study was to report on the incidence of biliary tract injuries in a single unit, comment on the way they were managed and look at strategies to prevent them. METHODS: A retrospective audit was conducted on laparoscopic cholecystectomies performed between January 1992 and March 2001. The data was collated from patient medical record files and yielded a total of 1216 procedures. RESULTS: There were 899 women (74%) and 317 men (26%), with an age range of 13-92 years. Most of the procedures were performed on an elective (94%) rather than emergent basis (6%). There was one bile duct injury (0.09%) and seven bile leaks (0.63%). The single injury involved common bile duct obstruction by a misplaced clip and was successfully managed by chol-angio-enteric bypass. Of the seven bile leaks, three were from the cystic duct stump, two from the gallbladder bed, and two were unidentified, settling conservatively. Of the five patients actively treated, two underwent therapeutic laparoscopy, two proceeded to laparotomy, and one was managed successfully by endoscopic stenting. CONCLUSIONS: Single-centre studies such as this are important in ensuring that standards of surgery are maintained in a community setting.

Adolescent↗
PubMed1986

Membrane-bound hydroxylases in elicitor-treated bean cells. Rapid induction of the synthesis of prolyl hydroxylase and a putative cytochrome P-450.

Treatment of cell-suspension cultures of bean (Phaseolus vulgaris cv. Canadian Wonder) with an elicitor preparation heat-released from the cell walls of the phytopathogenic fungus Colletotrichum lindemuthianum resulted in rapid changes in the activities of two microsomal oxygenases, cinnamic acid 4-hydroxylase, involved in accumulation of wall-bound phenolics and phytoalexins, and proline 2-oxoglutarate dioxygenase (prolyl hydroxylase) involved in the post-translational modification of hydroxyproline-rich glycoproteins. An anti-(cytochrome P-450) monoclonal antibody, originally raised against rat cytochrome P-450 isoform c, has been shown to bind to bean microsomes and recognise in Western blots an Mr-48,000 polypeptide, which comigrates with a haeme-containing protein on SDS/polyacrylamide gel analysis and which has been tentatively identified as a cytochrome P-450 capable of the hydroxylation of cinnamic acid. A preparation of proline 2-oxoglutarate dioxygenase purified to homogeneity was used to immunise rabbits for the production of antiserum. An elicitor-induced polypeptide of Mr 65,000 was identified as prolyl hydroxylase while an antigenically related polypeptide of Mr 60,000 was also immunoprecipitated but not induced by elicitor treatment. Use of the two antibodies has demonstrated rapid transient increases in the synthesis of the Mr 48,000 and Mr 65,000 oxygenases in vivo and for mRNAs as measured in in vitro translations, particularly for the putative cytochrome P-450. These increases slightly precede corresponding changes in the synthesis of the soluble enzyme phenylalanine ammonia-lyase, in common with which these oxygenases probably share a mechanism of gene activation underlying the increased activities seen in response to elicitor treatment.

Antibodies, Monoclonal↗
PubMed2003

Phaseolin in vitro pepsin digestibility: role of acids and phenolic compounds.

Great Northern bean (Phaseolus vulgaris L.) phaseolin proteolysis at 37 degrees C, varying HCl concentrations (10 mM to 1 M), phaseolin:pepsin ratios ranging from 5:1 to 100:1 (w/w), and incubation times up to 24 h was investigated. The results suggest that phaseolin is not resistant to in vitro pepsin hydrolysis. At a phaseolin-to-pepsin ratio of 100:1 (w/w), native phaseolin was completely digested in 24 h when incubated in 50 mM HCl, while heat-denatured phaseolin (30 min at 100 degrees C, boiling water bath) was digested in 1 h under similar conditions. When incubated at 37 degrees C for 24 h, acid alone, even at as low a concentration as 10 mM, caused a partial breakdown of native phaseolin. The degree of phaseolin hydrolysis by HCl was dependent on the acid concentration used. The rate of native phaseolin hydrolysis increased with increasing HCl concentration rather than pepsin concentration. Common food acids were able to partially hydrolyze phaseolin. Among the food acids tested, oxalic acid was the most effective in hydrolyzing phaseolin. Spectroscopic studies revealed a significant change in secondary and tertiary structures when native phaseolin was incubated in dilute HCl. None of the tested phenolic compounds adversely affected phaseolin hydrolysis by pepsin.

Chromatography↗
PubMed1982

Investigation of the Mechanism of Action of a Chlorosis-Inducing Toxin Produced by Pseudomonas phaseolicola.

A toxin that induced chlorotic haloes (typifying haloblight disease) on primary leaves of Phaseolus vulgaris L. (var. Canadian Wonder) was partially purified from culture filtrates of the causative agent Pseudomonas phaseolicola (Burkh.) Dowson. This material was used to investigate chlorosis induction. Haloes could only be induced in those bean leaves that were expanding and synthesizing chlorophyll (Chl); the toxin, therefore, does not promote Chl breakdown. Chl, carotene, and xanthophyll synthesis were inhibited in sections of greening barley (Hordeum vulgare L.) leaves, irrespective of the irradiance level. In parallel experiments, the toxin decreased the level of 5-aminolevulinic acid by amounts sufficient to account for toxin-inhibition of Chl synthesis. Electron microscopy revealed no difference between the transformation of etioplasts into chloroplasts in toxin-treated and control tissue, despite a 60% reduction in Chl in the former. The incorporation of [(14)C]acetate into lipid by greening barley leaf sections and by isolated Pisum sativum chloroplasts in the light and the dark was inhibited about 60% by the toxin. The distribution of radioactivity among the spectra of acyl residues was the same in the control and toxin-treated material. It is suggested that the toxin interferes with an early process common to the synthesis of different lipids, including Chl.

Journal Article↗