CHL1 encodes a component of the low-affinity nitrate uptake system in Arabidopsis and shows cell type-specific expression in roots

The Arabidopsis CHL1 (AtNRT1) gene confers sensitivity to the herbicide chlorate and encodes a nitrate-regulated nitrate transporter. However, how CHL1 participates in nitrate uptake in plants is not yet clear. In this study, we examined the in vivo function of CHL1 with in vivo uptake measurements and in situ hybridization experiments. Under most conditions tested, the amount of nitrate uptake by a chl1 deletion mutant was found to be significantly less than that of the wild type. This uptake deficiency was reversed when a CHL1 cDNA clone driven by the cauliflower mosaic virus 35S promoter was expressed in transgenic chl1 plants. Furthermore, tissue-specific expression patterns showed that near the root tip, CHL1 mRNA is found primarily in the epidermis, but further from the root tip, the mRNA is found in the cortex or endodermis. These results are consistent with the involvement of CHL1 in nitrate uptake at different stages of root cell development. A functional analysis in Xenopus oocytes indicated that CHL1 is a low-affinity nitrate transporter with a K(m) value of approximately 8.5 mM for nitrate. This finding is consistent with the chlorate resistance phenotype of chl1 mutants. However, these results do not fit the current model of a single, constitutive component for the low-affinity uptake system. To reconcile this discrepancy and the complex uptake behavior observed, we propose a "two-gene" model for the low-affinity nitrate uptake system of Arabidopsis.     

Prenatal determination of human platelet antigen type using DNA amplification following amniocentesis

OBJECTIVE: To assess the effectiveness of a newly developed individualised birthweight ratio (IBR), which corrects for physiological birthweight determinants, in identifying infants at risk from the complications of macrosomia. DESIGN: Prospective observational study. SETTING: Obstetric unit, Nottingham City Hospital. SUBJECTS: 2835 women delivered between December 1991 and July 1992 and the infants of 624 of these, selected by virtue of their birthweight for gestation and IBR centile positions. MAIN OUTCOME MEASURES: Skinfold thickness and ponderal index measurements, operative delivery, shoulder dystocia, fetal trauma, impaired glucose tolerance. RESULTS: Using an IBR above the 90th centile as a cut off results in 2.4% of infants being reclassified as normally grown and 3.1% are reclassified as large. The IBR does not result in the identification of any more infants with abnormal ponderal indices or skinfold thicknesses than birthweight for gestation. It does, however, identify more of the infants at risk of operative delivery, shoulder dystocia, fetal trauma and impaired glucose tolerance. CONCLUSION: The IBR significantly improves upon birthweight for gestation in identifying infants who suffer from the complications of relative macrosomia.     

Treatment with anti-interleukin-10 monoclonal antibody enhances early resistance to but impairs complete clearance of Listeria monocytogenes infection in mice

Mice that received an anti-interleukin-10 (anti-IL-10) neutralizing monoclonal antibody (MAb) (SXC-1) prior to infection with Listeria monocytogenes initially demonstrated resistance to the infection, as indicated by reduced recovery of L. monocytogenes from their spleens and livers during the first 5 days after challenge. Anti-IL-10 MAb-treated mice then demonstrated reduced resistance during the later stage of infection, as indicated by persistent infection with L. monocytogenes in their livers 11 days after challenge. Aspartate aminotransferase (AST) levels (a measure of liver damage) in the sera of control mice increased between 1 and 5 days after challenge, while anti-IL-10 MAb-treated mice maintained lower AST levels. At 7 days after challenge, AST levels in the sera of control mice decreased as the numbers of organisms declined. In contrast, AST levels increased as the infections persisted in anti-IL-10 MAb-treated mice. The AST levels in serum reflected liver histopathology as anti-IL-10 MAb-treated mice exhibited fewer granulomatous lesions and less necrosis of liver tissue than the control mice during the first 5 days after challenge. Anti-IL-10 MAb treatment altered the expression of inflammatory cytokine mRNAs during L. monocytogenes infection. Control MAb-treated mice exhibited increased expression of tumor necrosis factor alpha and granulocyte-macrophage colony-stimulating factor mRNA in their lives during L. monocytogenes infection, but this increase did not occur in anti-IL-10 MAb-treated mice. Gamma interferon mRNA expression in the livers of the control MAb-treated mice was increased between 1 and 5 days after L. monocytogenes challenge and then decreased at 7 days after challenge. In contrast, gamma interferon mRNA expression in the livers of anti-IL-10 MAb-treated mice was not decreased until 7 days after challenge. These results indicate that endogenous IL-10 has both beneficial and detrimental effects on the host response to L. monocytogenes infection in mice.     

Composition of electrodeposited Zn–Cr alloy coatings and phase transformations induced by thermal treatment

Zn–Cr alloy coatings were obtained in a flow cell, for modeling the process of high speed electrodeposition on steel strips. Alloy coatings, containing between 6 and 18 at.% Cr were annealed at 260 °C in an inert atmosphere. The phase composition and the crystallographic characteristics of “as prepared” and “annealed” coatings, were studied by XRD. It is shown that non-equilibrium δ- and Γ-(Zn,Cr) phases are major constituents of the “as prepared” coatings. On annealing, equilibrium ζ-CrZn₁₃ phase precipitates from δ- and Γ-supersaturated solid solutions. The lattice parameters and the similarities in phase composition of the annealed coatings, deposited onto two types of substrates – low carbon steel and Cr plated (protected) low carbon steel – show that if Fe from the substrate “contaminates” the precipitated ζ-CrZn₁₃ phases, its relative amount do not exceed few tenths of a percent.The influence of the elemental composition, conditions of electrochemical deposition, and post-deposition thermal treatment on phase composition of the coatings is discussed.     

Role of paired basic residues in the expression of active recombinant galactosyltransferases from the bacterial pathogen Neisseria meningitidis

The lgtB gene encoding a beta-1,4-galactosyltransferase gene and the lgtC gene encoding an alpha-1,4-galactosyltransferase from the bacterial pathogen Neisseria meningitidis were cloned into an expression vector and overexpressed in Escherichia coli. Both genes expressed very well, but problems with C-terminal proteolysis were encountered with both proteins. The lgtC protein was initially isolated from extracts of recombinant E.coli as a truncated species that retained enzymatic activity, and was subsequently shown by mass spectrometry to be 19 residues shorter than the expected protein. A specific set of engineered C-terminal deletions was constructed to investigate their effect on the expression of lgtC. As many as 28 residues could be deleted with little effect on activity, and with the concomitant improvement of the overall expression up to fivefold over the full length protein. The lgtB protein was also proteolysed in extracts of normal E.coli strains into enzymatically inactive fragments lacking 28 or 41 C-terminal residues. This degradation could be prevented by expression in an ompT protease deficient strain of E.coli. The full length lgtB protein was not stable in soluble protein extracts from all recombinant strains, however a stable enzyme preparation could be achieved with the membrane fraction from cells of the ompT deficient strain expressing lgtB. Specific deletions of lgtB were also constructed, and 15 residues could be removed without loss of enzyme activity and also with the concomitant improvement of the overall expression up to twofold over the full length protein. Longer deletions produced protein but activity could not be detected in these recombinant strains. Examination of the glycosyltransferase sequences from a wide range of bacteria showed their C-terminal segments of approximately 50 amino acids frequently contained paired basic residues. Engineering of these segments may therefore be required as a general practice to produce these enzymes for use in the large scale chemi-enzymatic synthesis of carbohydrate-based therapeutics.      

PARTICLE STIMULATED NUCLEATION AND THE FORMATION OF RSCRYSTALLIZATION TEXTURE IN Al-Mn ALLOY CONTAINING PARTICLES Ⅱ. Interaction Between Particles and Other Nucleation Sites and the Formation of Recrystallization Texture

本文报了Ａｌ－Ｍｎ合金形昧形变取向基体中粒子周围亚晶转动规律及对应的粒子促进形核晶粒的取向特征的基础上,进一步测定了处在晶界,立方带上的了周转新晶粒的取向分布。     

Carcinoembryonic antigen in diagnosis and monitoring of colorectal cancer]

Female SJL mice are more susceptible than male mice to experimental autoimmune encephalomyelitis (EAE) induced by myelin basic protein (MBP)-specific T lymphocytes. In the present study, we examined mechanisms involved in this gender-related difference in disease susceptibility. MBP-specific T lymphocytes derived from spleens of males during the effector phase of adoptive EAE produced significantly higher levels of IL-10, an anti-inflammatory cytokine in EAE. A protective effect of testosterone was then shown. Females implanted with dihydrotestosterone pellets demonstrated a significantly less severe course of EAE as compared with females implanted with placebo pellets. Finally, MBP-specific T lymphocytes derived from dihydrotestosterone-implanted females produced significantly higher levels of IL-10 than those from placebo. Together these data indicate that testosterone exerts a protective effect in EAE that is mediated at least in part by enhanced production of IL-10 by autoantigen-specific T lymphocytes.