The cellular retinoic-acid-binding protein is expressed in tissues associated with retinoic-acid-induced malformations

Retinoic acid (RA) is thought to play a role in embryonic pattern formation in vertebrates. A naturally occurring gradient of endogenous RA has been demonstrated in the developing chick limb bud, while local application of RA leads to the formation of additional digits. In mammals, a well-defined spectrum of birth defects has been reported as a result of fetal exposure to excess RA. In analogy to the chick limb bud, it may be speculated that these malformations are the result of disturbance of morphogenetic RA concentration gradients. A candidate gene involved in the regulation of endogenous RA concentrations is the gene encoding cellular RA binding protein (CRABP). We have isolated a partial cDNA clone corresponding to the chicken homolog of CRABP, and performed in situ hybridization experiments on sections of embryos at various stages of development. CRABP expression was detected in the CNS, the craniofacial mesenchyme, ganglia of the peripheral nervous system, the limb bud, and the visceral arch area. Our results indicate that the spatiotemporally specified expression pattern displayed by the CRABP gene exhibits a striking correspondence to the tissues that are affected by exposure of avian or mammalian embryos to RA. We hypothesize that CRABP plays an important role in normal embryogenesis and that embryonic tissues showing high CRABP expression are susceptible to the adverse effects of excess RA.     

Identification of loci associated with tolerance to Johne’s disease in Holstein cattle

Johne’s disease, caused by Mycobacterium avium subspecies paratuberculosis (Map), is a fatal disease in cattle. The objective of this study was to identify loci associated with tolerance in cows infected with Map. Tolerance was defined as a cow’s fitness at a given level of Map infection intensity. Fitness was measured by Map faecal cultures, and Map infection intensity was measured by culturing four gut tissues. The quantitative phenotype of tolerance was defined by numerical indexes of cultures of peak (peak tolerance, PT) and average (average tolerance, AT) faecal and tissue Map from 245 Holstein cows. The categorical phenotype was defined as: ≥100 cfu Map tissue infection, and faecal shedding ≥75 cfu (intolerant) or <10 cfu (tolerant cows). In 94 cows, Map was identified in ≥1 tissue, including 44 cows with ≥100 Map tissue cfu and 36 with ≥1 faecal cfu. A genome‐wide association analysis was performed after filtering, leaving genotypes for 45 789 SNPs in 90 animals for the quantitative phenotype and 16 cases and 25 controls for the categorical analysis of tolerance. rs41748405:A>C (BTA15) was associated with PT (P = 1.12 × 10^?7) and AT (P = 2.17 × 10^?6). Associations were identified with PT and adjacent SNPs ss61512613:A>G and ss61530518:A>G (BTA6) (P < 3.0 × 10^?5), and with AT for ss61469568:A>G (BTA 2) (P = 3.3 × 10^?5) and ss86284768:A>G (BTA1) (P = 3.31 × 10^?5). For the categorical phenotype, an association was found with ss8632653:A>G (BTA6) (P < 5.0 × 10^?5). This is the first study to identify loci associated with tolerance to Johne’s disease.     

Chemotaxis inhibitory protein of Staphylococcus aureus binds specifically to the C5a and formylated peptide receptor

Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is an exoprotein produced by several strains of S. aureus, and a potent inhibitor of neutrophil and monocyte chemotaxis toward C5a and formylated peptides like fMLP. These chemoattractants act on their target cells by binding and activating the C5aR and formylated peptide receptor (FPR), respectively. In the present report, we examined the mechanism by which CHIPS affects both of these receptors. We showed that CHIPS blocked binding of anti-C5aR mAb and formylated peptide to human neutrophils as efficiently at temperatures of 0 and 37 degrees C, implying that it is independent of signal transducing systems. This was confirmed by showing that CHIPS acts completely independently of ATP. Additionally, CHIPS was not internalized upon binding to neutrophils. Furthermore, we showed that CHIPS binds specifically to the C5aR and FPR expressed on U937 cells. This binding was functional in blocking C5a- and fMLP-induced calcium mobilization in these cell lines. These results suggest that CHIPS binds directly to the C5aR and FPR, thereby preventing the natural ligands from activating these receptors. The apparent K(d) values of CHIPS for the C5aR and FPR were 1.1 +/- 0.2 nM and 35.4 +/- 7.7 nM, respectively. Moreover, after screening a wide variety of other G protein-coupled receptors, CHIPS was found to affect exclusively the C5aR and FPR. This selectivity and high-affinity binding with potent antagonistic effects makes CHIPS a promising lead for the development of new anti-inflammatory compounds for diseases in which damage by neutrophils plays a key role.     

A GDP-fucose:[Gal beta 1----4]GlcNAc alpha 1----3-fucosyltransferase activity is correlated with the presence of human chromosome 11 and the expression of the Lex, Ley, and sialyl-Lex antigens in human-mouse cell hybrids

By fusion of human leukocytes and cells of the murine myeloid cell line WEHI-TG, we produced human-mouse myeloid cell hybrids. Hybrids which contain human chromosome 11 have been demonstrated to express the myeloid-associated carbohydrate antigen Lex (Geurts van Kessel, A. H. M., Tetteroo, P. A. T., Von dem Borne, A. E. G. Kr., Hagemeijer, A., and Bootsma, D. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 3748-3752). In this paper, we report that the hybrids that contain chromosome 11 also expressed the Lex-related antigens Ley and sialyl-Lex. Glycosyltransferase activities were measured in a panel of six such hybrid cell lines, and the correlation to antigen expression and to the presence of human chromosomes was investigated. GDP-fucose:[Gal beta 1----4]GlcNAc alpha 1----3-fucosyltransferase activity in the hybrids tested correlated with the expression of Lex, Ley, and sialyl-Lex and with the occurrence of chromosome 11. No such correlation was found for several other glycosyltransferases involved in the synthesis of these antigens. These findings suggest that the gene for alpha 3-fucosyltransferase is located on chromosome 11 and that it is through the activity of this enzyme that the expression of Lex, Ley, and sialyl-Lex in human myeloid cells is regulated.     

Impact of elevated CO2 on soil organic matter dynamics as related to changes in aggregate turnover and residue quality

Increasing global atmospheric CO₂ concentration can potentially affect C cycling in terrestrial ecosystems. This study was conducted to assess the impact of elevated CO₂ concentration on soil organic matter and aggregate dynamics in Lolium perenne and Trifolium repens pastures. Soil samples from a 6 year old `free air CO₂ enrichment' (FACE) experiment were separated in four aggregate size classes (<53, 53–250, 250–2000, and > 2000 m). Free light fraction (i.e. particulate organic matter (POM) outside of aggregates; free LF) and intra-aggregate-POM (i.e. POM occluded within the aggregate structure; iPOM) were isolated. The distinct ¹³C-signature of the CO₂ used to raise the ambient CO₂ concentration in FACE allowed us to calculate proportions of recently incorporated C (< 6 yr) in the physically defined soil fractions. The proportion of new C increased with increasing aggregate size class, except the two largest aggregate size classes had a similar proportion of new C; this indicates a faster turnover of macroaggregates compared to microaggregates. In addition, higher proportions of new C in macroaggregates under T. repens compared to L. perenne indicate a faster macroaggregate turnover under T. repens. This faster macroaggregate turnover is hypothesized to be a result of the higher residue quality (C:N ratio) of T. repens compared to L. perenne and reduces the potential of sequestering C under elevated CO₂. In the L. perenne soil, elevated CO₂ did not significantly increase total C, but led to: (1) a 54% increase in aggregation and (2) a 40% increase in total iPOM-C. It is hypothesized that the sequestration of iPOM-C induced by elevated CO₂ in the low residue quality, L. perenne treatment, resulted from an increase in the proportion of large macroaggregates with a slow turnover.     

Optimizing rice yields while minimizing yield‐scaled global warming potential

To meet growing global food demand with limited land and reduced environmental impact, agricultural greenhouse gas (GHG) emissions are increasingly evaluated with respect to crop productivity, i.e., on a yield‐scaled as opposed to area basis. Here, we compiled available field data on CH₄ and N₂O emissions from rice production systems to test the hypothesis that in response to fertilizer nitrogen (N) addition, yield‐scaled global warming potential (GWP) will be minimized at N rates that maximize yields. Within each study, yield N surplus was calculated to estimate deficit or excess N application rates with respect to the optimal N rate (defined as the N rate at which maximum yield was achieved). Relationships between yield N surplus and GHG emissions were assessed using linear and nonlinear mixed‐effects models. Results indicate that yields increased in response to increasing N surplus when moving from deficit to optimal N rates. At N rates contributing to a yield N surplus, N₂O and yield‐scaled N₂O emissions increased exponentially. In contrast, CH₄ emissions were not impacted by N inputs. Accordingly, yield‐scaled CH₄ emissions decreased with N addition. Overall, yield‐scaled GWP was minimized at optimal N rates, decreasing by 21% compared to treatments without N addition. These results are unique compared to aerobic cropping systems in which N₂O emissions are the primary contributor to GWP, meaning yield‐scaled GWP may not necessarily decrease for aerobic crops when yields are optimized by N fertilizer addition. Balancing gains in agricultural productivity with climate change concerns, this work supports the concept that high rice yields can be achieved with minimal yield‐scaled GWP through optimal N application rates. Moreover, additional improvements in N use efficiency may further reduce yield‐scaled GWP, thereby strengthening the economic and environmental sustainability of rice systems.     

Distribution of viruses in the Chesapeake Bay.

High virus counts were found in water samples collected from the Chesapeake Bay. Viruses were enumerated by ultracentrifugation of water samples onto grids which were visualized by transmission electron microscopy. Virus counts in September 1990, April 1991, June 1991, August 1991, and October 1991 ranged between 2.6 x 10(6) and 1.4 x 10(8) viruses ml-1 with a mean of 2.5 x 10(7) viruses ml-1. Virus counts were usually at least three times higher than direct bacterial counts in corresponding samples. Virus counts in August and October were significantly higher than at the other sampling times, whereas bacterial counts were significantly lower at that time, yielding mean virus-to-bacterium ratios of 12.6 and 25.6, respectively. From analysis of morphology of the virus particles, it is concluded that a large proportion of the viruses are bacteriophages. The high virus counts obtained in this study suggest that viruses may be an important factor affecting bacterial populations in the Chesapeake Bay, with implications for gene transfer in natural aquatic bacterial populations and release of genetically engineered microorganisms to estuarine and coastal environments.     

Recombineering mycobacteria and their phages

Bacteriophages are central components in the development of molecular tools for microbial genetics. Mycobacteriophages have proven to be a rich resource for tuberculosis genetics, and the recent development of a mycobacterial recombineering system based on mycobacteriophage Che9c-encoded proteins offers new approaches to mycobacterial mutagenesis. Expression of the phage exonuclease and recombinase substantially enhances recombination frequencies in both fast- and slow-growing mycobacteria, thereby facilitating construction of both gene knockout and point mutants; it also provides a simple and efficient method for constructing mycobacteriophage mutants. Exploitation of host-specific phages thus provides a general strategy for recombineering and mutagenesis in genetically naive systems.     

INTRANUCLEAR AND CYTOPLASMIC ANNULATE LAMELLAE IN TUNICATE OOCYTES

Electron microscope studies were made on various tunicate oocytes at different stages of growth and development. Both the inner and outer lamellae of the perforated nuclear envelope demonstrate considerable blebbing activity. The blebs of the inner lamella detach into the nucleoplasm where they undergo a special type of fusion process resulting in the formation of numerous, usually single, differentiated annulate lamellae of various lengths. The blebbing of the outer layer of the nuclear envelope contributes to the vesicular and granular endoplasmic reticulum characteristically present in the ooplasm and perhaps to the differentiation of cytoplasmic annulate lamellae as well. Cytoplasmic stacks of annulate lamellae frequently have ribosomes associated with them. In addition, granular accumulations are sometimes observed around or between the annuli. The morphological evidence suggests that, at least in many cases, the annuli in the annulate lamellae are patent.     

Isolation and characterization of a generalized transducing phage for Xanthomonas campestris pv. campestris.

We have isolated and characterized a lytic double-stranded DNA Xanthomonas campestris pv. campestris bacteriophage (XTP1) capable of mediating generalized transduction. The phage transduces chromosomal markers at frequencies of 10(-5) to 10(-6) transductants per PFU. We demonstrated its genetic utility by the isolation and cotransduction of linked transposon insertions to a nonselectable locus, xgl, required for the cleavage of 5-bromo-3-chloro-indoyl-beta-D-galactoside and showed that rif and str alleles in X. campestris are 75% linked. One-step growth experiments showed that the latent and rise periods were each 2 h and the average burst size was 35. The DNA genome is approximately 180 kb, presumably modified in a sequence-specific manner, and may be covalently attached to protein(s). Electron micrographs show the phage particle to have an icosahedral head and contractile tail with tail fibers uniquely attached to a location 40 nm proximal from the end of the tail.