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Increased expression of preprotachykinin, calcitonin gene-related peptide, but not vasoactive intestinal peptide messenger RNA in dorsal root ganglia during the development of adjuvant monoarthritis in the rat.

Neuropeptides in dorsal root ganglia (DRG) have been implicated in the pathogenesis of pain and neurogenic inflammation in experimental and clinical arthritis. Recently we demonstrated increased levels of substance P (SP) and calcitonin gene-related peptide (CGRP) confined to innervating DRG in adjuvant-mediated monoarthritis. We have now investigated whether changes in peptide content are reflected in altered neuropeptide gene expression and the time course involved. Using in situ hybridization we found marked increases in expression of beta-preprotachykinin (PPT; 81 +/- 24% rise) and alpha-CGRP (44 +/- 6% rise) mRNAs in innervating (ipsilateral L5) DRG neurones only. These increases occurred at the onset of acute inflammation (8 h) and persisted until chronic arthritis developed after 14 days. There were no changes in the proportion of DRG neurones expressing PPT or CGRP mRNAs. Messenger RNA encoding vasoactive intestinal polypeptide (VIP) was not induced. These data suggest that increased synthesis of PPT and CGRP peptides in DRG may play a role in the pathogenesis both of adjuvant-mediated acute inflammation and chronic arthritis.

Animals↗

Expression of high-molecular-weight neurofilament protein and vimentin in human developing spinal cord and dorsal root ganglia.

Expression of intermediate filament proteins in developing human dorsal root ganglia and spinal cord was studied using double-label immunofluorescence on frozen sections with monoclonal antibodies to vimentin and high-molecular-weight neurofilament protein (NF-H). Both proteins were found in the nerve fibres inside the ganglia of 6- and 8-week embryos. In 9-14-week embryos the NF-H protein was found in the perikarya of ganglion cells. Vimentin was only sporadically present in these cells, but was accumulated in connective tissue cells. In the following development the NF-H protein expression was limited to the ganglion cells and processes, and vimentin was confined to the satellite cells and mesenchymal derivatives. In the spinal cord of the 6-week and 8-week human embryos NF-H protein was found in the longitudinal fibres of the marginal layer and in a loose network of nerve fibres in the mantle zone. Some of the nerve fibres were also vimentin positive. The coexpression of both proteins stopped during weeks 9-10. In 30-week foetuses vimentin was only found in glial cells and mesenchymal derivatives, while NF-H protein was in the axons of both white and grey matter but not in neuronal perikarya.

Antibodies, Monoclonal↗

Organization and cell differentiation in lateral roots of Arabidopsis thaliana.

Lateral root formation in plants involves the stimulation of mature pericycle cells to proliferate and redifferentiate to create a new organ. The simple organization of the root of Arabidopsis thaliana allows the development of lateral root primordia to be characterized histologically. We have divided the process of lateral root development into 8 stages defined by specific anatomical characteristics and cell divisions. To identify the cell types in the developing primordium we have generated a collection of marker lines that express beta-glucuronidase in a tissue- or cell type-specific manner in the root. Using these tools we have constructed a model describing the lineage of each cell type in the lateral root. These studies show that organization and cell differentiation in the lateral root primordia precede the appearance of a lateral root meristem, with differential gene expression apparent after the first set of divisions of the pericycle.

Arabidopsis↗

The development of the dorsal root potential and the responsiveness of primary afferent fibers to gamma-aminobutyric acid in the spinal cord of rat fetuses.

The development of the dorsal root potential (DRP) and the responsiveness of primary afferent fibers to gamma-aminobutyric acid (GABA) were investigated in the isolated spinal cord of rat fetuses. At embryonic day 15.5, stimulation of the lumbar dorsal root was first effective in eliciting the DRP, which was not inhibited by bicuculline. A bicuculline-sensitive component of the DRP appeared at embryonic day 17.5. GABA (10 microM to 1 mM) caused a dose-dependent depolarization of the primary afferent fibers from embryonic day 13.5. The amplitude of the depolarization gradually increased with age until embryonic day 17.5 and was maintained thereafter. If the bicuculline-sensitive DRP solely reflects GABAergic activity, it is suggested that GABAergic activity develops at embryonic day 17.5 and the development of the responsiveness of primary afferent fibers to GABA precedes the functional onset of GABAergic neurons.

Animals↗

Hedgehog pathway gene expression during early development of the molar tooth root in the mouse.

Sonic hedgehog is a secreted protein important for many aspects of embryonic development. In the developing tooth, Shh expression is restricted to the epithelial compartment and plays an important role during both initiation and subsequent coronal morphogenesis. We have investigated the expression of Shh and constituent members of the signalling pathway during early development of the molar tooth root in the mouse and find the presence of transcripts in Hertwig's epithelial root sheath. These epithelial cells of the root sheath and the surrounding apical mesenchyme of the dental papilla and follicle also expressed the Shh receptor Ptc1, agonist Smo and Gli downstream transcriptional effectors; however, this response occurred over short range. In contrast, the Shh antagonists Hip1 and Gas1 were both expressed at a distance from these responding cells, in more peripheral regions of the developing root. Transcripts of the Skn acyl transferase lacked specific expression in early root structures.

Animals↗

Sugar-induced adventitious roots in Arabidopsis seedlings.

The effects of sugars on root growth and on development of adventitious roots were analyzed in Arabidopsis thaliana. Seeds were sown on agar plates containing 0.0-5.0% sugars and placed vertically in darkness (DD) or under long day (LD, 16 h:8 h) conditions, so that the seedlings were constantly attached to the agar medium. In the sucrose-supplemented medium, seedlings showed sustained growth in both DD and LD. However, only dark-grown seedlings developed adventitious roots from the elongated hypocotyl. The adventitious roots began to develop 5 days after imbibition and increased in number until day 11. They could, however, be initiated at any position along the hypocotyl, near the cotyledon or the primary root. They were initiated in the pericycle in the same manner as ordinary lateral roots. Sucrose, glucose and fructose greatly stimulated the induction of adventitious roots, but mannose or sorbitol did not. Sucrose at concentrations of 0.5-2.0% was most effective in inducing adventitious roots, although 5.0% sucrose suppressed induction. Direct contact of the hypocotyl with the sugar-supplemented agar medium was indispensable for the induction of adventitious roots.

Arabidopsis↗

Mycorrhizal colonization of transgenic aspen in a field trial.

Mycorrhizal colonization of genetically modified hybrid aspen (Populus tremula x P. tremuloides Michx.) was investigated over 15 months in a field experiment. The aspen carried the rolC gene from Agrobacterium rhizogenes under control of either the constitutive cauliflower mosaic virus 35S promoter or the light-inducible rbcS promoter. Arbuscular mycorrhizas (AMs) were rare in all root samples, while fully developed ectomycorrhizas (EMs) were found in all samples. No significant differences in the degree of mycorrhizal colonization between aspen lines were seen with either AMs or EMs. The EM community on the release area was dominated by four fungal species that formed more than 90% of all mycorrhizas, while eleven EM types were found occasionally. Mycorrhizal diversity did not differ between transgenic and non-transgenic trees. The structure of mycorrhizal communities was similar for most aspen lines. The sole significant difference was found in the abundance and development of one of the four common EM morphotypes, which was rare and poorly developed on roots from the transgenic aspen line Esch5:35S-rolC-#5 compared with non-transgenic controls. This effect is clone specific as the formation of this EM type was not affected by the transgene expression in the other transgenic line, Esch5:35S-rolC-#1. This is the first demonstration of a clonal effect influencing the ability of a transgenic plant to form a mycorrhizal symbiosis with a potential fungal partner.

Fungi↗

Efficient whole plant regeneration of common bean (Phaseolus vulgaris L.) using thin-cell-layer culture and silver nitrate.

A method was designed to optimize rapid and high frequency direct shoot regeneration (without intermediate callus) of the commercially important common bean, Phaseolus vulgaris L., using the transverse thin cell layer (tTCL) method. The pretreatment of seeds with 10 µM TDZ significantly increased bud regeneration frequency on tTCL. A 2-week culture of tTCLs on 10 µM TDZ followed by a reduction in the TDZ concentration (1 µM) was needed to achieve optimal bud induction and further development of the neo-formed buds. An incubation period greater than 2 weeks of tTCLs with 10 µM TDZ concentration resulted in inhibitory effects on the development of shoots and roots. Shoot development was enhanced by 10 µM BAP and 10 µM AgNO(3) leading to 100% well developed shoots. Regenerated plants developed into true-to-type fertile plants.

Journal Article↗

Two genes encoding extension-like proteins are predominantly expressed in tomato root hair cells.

A differential screen of a tomato root hair cDNA library resulted in the cloning of two cDNAs, Dif10 and Dif54, whose corresponding genes are preferentially expressed in root hair cells as determined by analysis of mRNA levels in various tomato organs. Transcript levels showed no increase in leaves subjected to hormonal and environmental stress treatments. Sequence analysis of the cDNAs revealed high similarity to members of the extension family. Extensions are hydroxyproline-rich glycoproteins (HRGPs) located in the cell wall. In order to study the functional significance of HRGPs in root hairs, tomato seedling roots were treated with micromolar concentrations of 3,4-dehydro-L-proline (Dhp), a selective inhibitor of prolyl hydroxylase. Dhp treatment resulted in changes in root growth and the development of root hairs with reduced hair length, suggesting an important role of HRGPs in hair morphogenesis.

Amino Acid Sequence↗

Early and late root formation in epicotyl cuttings of Pinus sylvestris after auxin treatment.

Auxin stimulated rooting of epicotyl cuttings of Pinus sylvestris irrespective of the physiological stage of the plant from which the cuttings were taken. The epicotyl cuttings were divided into two groups, those that differentiated roots early (within 6 weeks after cutting) and those that differentiated roots late (more than 6 weeks after cutting). Auxin treatment significantly stimulated the frequency of early rooting of epicotyl cuttings, whereas it had no effect on the frequency of late rooting of epicotyl cuttings. The number of roots per rooted cutting was significantly higher after auxin treatment both on early and late rooted cuttings. Anatomical studies showed that most of the roots on both early and late rooted cuttings developed from wound tissue. However some types of rooting were found only after auxin treatment of early rooted cuttings, e.g., roots that developed from resin duct wound tissue or from vascular tissue or in the pith.

Journal Article↗

Induction of lateral root structure formation on petunia roots: A novel effect of GMI1000 Ralstonia solanacearum infection impaired in Hrp mutants.

Ralstonia solanacearum is a soilborne plant pathogen that invades its host via roots. As in many gram-negative bacterial plant pathogens, the R. solanacearum Hrp type III secretion system is essential for interactions of the bacterium with plants; however, the related mechanisms involved in disease expression are largely unknown. In this work, we examined the effects of infection by R. solanacearum GMI1000 and Hrp mutants on the root system of petunia plants. Both the wild-type and mutant strains disturbed the petunia root architecture development by inhibiting lateral root elongation and provoking swelling of the root tips. In addition, GMI100 but not the Hrp mutants induced the formation of new root lateral structures (RLS). This ability is shared by other, but not all, R. solanacearum strains tested. Like lateral roots, these new structures arise from divisions of pericycle founder cells which, nevertheless, exhibit an abnormal morphology. These RLS are efficient colonization sites allowing extensive bacterial multiplication. However, they are not required for the bacterial vascular invasion that leads to the systemic spread of the bacterium through the whole plant, indicating that, instead, they might play a role in the rhizosphere-related stages of the R. solanacearum life cycle.

Bacterial Proteins↗

Clonal propagation of softwoods.

Softwoods or gymnosperms, which make up 60% of the forested areas of the world, are economically important as a source of lumber, pulp, and paper. Reforestation is a major activity worldwide and the potential benefits of using clonal planting stock have long been recognized. Tissue culture clonal methods or micropropagation is a newer approach that can be achieved by enhancing axillary bud breaking, production of adventitious buds (organogenesis), and somatic embryogenesis. Plantlet production via organogenesis requires at least four stages: (1) bud induction on the explant, (2) shoot development and multiplication, (3) rooting of developed shoots, and (4) hardening of plantlets. Similarly, the production of plantlets via somatic embryogenesis, which has the potential to produce a larger number of plantlets, and in a shorter period of time, also requires several stages. These include (1) induction, maintenance, and proliferation of embryogenic tissue; (2) maturation (both morphological and physiological) of somatic embryos; and (3) germination and conversion of the somatic embryos. In this chapter, plantlet production via organogenesis from seedling and adolescent/mature explants and somatic embryogenesis from immature and mature seeds of white spruce (Picea glauca) are outlined.

Culture Media↗

A histopathological study of various root canal filling materials.

This report compares tissue irritation by a newly developed calcium-based root canal filling material with irritation by previously available materials and evaluates the toxicity of various components in the root canal filling material. Polyethylene tubes filled with root canal filling materials were implanted in rat subcutaneous tissue, enucleated after 2, 4, 8 and 12 weeks, and histopathologically evaluated for tissue reaction. The tissue inflammation was microscopically evaluated and divided into 4 grades. The new root canal filling material has good tissue affinity compared to prior material. Transmission electronic microscopy and X-ray analysis revealed: titanium in AH-26; barium and bismuth in Canals, Tubli-seal and the newly developed root canal filling cement A; and zinc in the root canal filling cement B. Tissue was variously inflamed and, occasionally, necrosis was induced in the cells. The newly developed root canal filling cement A, showed better tissue affinity and no or slightly morphology change.

Animals↗

Functional conservation of a root hair cell-specific cis-element in angiosperms with different root hair distribution patterns.

Vascular plants develop distinctive root hair distribution patterns in the root epidermis, depending on the taxon. The three patterns, random (Type 1), asymmetrical cell division (Type 2), and positionally cued (Type 3), are controlled by different upstream fate-determining factors that mediate expression of root hair cell-specific genes for hair morphogenesis. Here, we address whether these root hair genes possess a common transcriptional regulatory module (cis-element) determining cell-type specificity despite differences in the final root hair pattern. We identified Arabidopsis thaliana expansinA7 (At EXPA7) orthologous (and paralogous) genes from diverse angiosperm species with different hair distribution patterns. The promoters of these genes contain conserved root hair-specific cis-elements (RHEs) that were functionally verified in the Type-3 Arabidopsis root. The promoter of At EXPA7 (Type-3 pattern) also showed hair cell-specific expression in the Type 2 rice (Oryza sativa) root. Root hair-specific genes other than EXPAs also carry functionally homologous RHEs in their promoters. The RHE core consensus was established by a multiple alignment of functionally characterized RHEs from different species and by high-resolution analysis of At EXPA7 RHE1. Our results suggest that this regulatory module of root hair-specific genes has been conserved across angiosperms despite the divergence of upstream fate-determining machinery.

Arabidopsis Proteins↗

Cell surface glycoconjugates and carbohydrate-binding proteins: possible recognition signals in sensory neurone development.

Dorsal root ganglion (DRG) neurones transmit cutaneous sensory information from the periphery to the dorsal horn of the spinal cord. Subpopulations of DRG neurones that subserve distinct sensory modalities project to discrete regions in the dorsal horn. The formation of specific sensory connections during development may involve cell-surface interactions with spinal cord cells. Molecules that are expressed on the surface of functional subpopulations of DRG and dorsal horn neurones have therefore been identified. Distinct subsets of DRG neurones express globo- or lactoseries carbohydrate differentiation antigens. The expression of defined carbohydrate structures correlates with the embryonic lineage, peptide phenotype and the central termination site of DRG neurones. Similar or identical glycoconjugates have been implicated in cellular interactions that contribute to preimplantation embryonic development. Small-diameter DRG neurones that project to the superficial dorsal horn express N-acetyllactosamine backbone structures that are potential ligands for beta-galactoside-specific binding proteins (lectins). Two lectins have been identified that are expressed early in development in the superficial dorsal horn. These complementary molecules may contribute to the development of sensory afferent projections in the spinal cord.

Aging↗

Mutans streptococci in subgingival plaque of treated and untreated patients with periodontitis.

BACKGROUND: The etiology of root caries is thought to be identical to coronal caries, though root caries seem to be more complicated because of the higher susceptibility of exposed roots (dentin) by periodontal therapy to demineralization than intact enamel. This implies that mutans streptococci are the most likely pathogens in the development of root caries. Although it is known that both the numbers of mutans streptococci and the frequency of isolation in root caries lesions are negatively correlated with the distance from the gingival margin, the subgingival sulcus has not been considered a possible habitat for mutans streptococci. However, subgingival occurrence of mutans streptococci in both untreated and treated periodontal patients has not been documented well in the literature. OBJECTIVE: To investigate the presence and levels of mutans streptococci in the subgingival plaque of patients (n=154) in different stages of periodontal therapy. The subgingival sulcus may be a possible habitat for mutans streptococci. This localisation of mutans streptococci may be of importance in the development of root caries after periodontal surgery. MATERIALS AND METHODS: In this cross-sectional study, subgingival plaque samples from 154 consecutive adult periodontitis patients were tested for presence and levels of mutans streptococci and putative periodontal pathogens by anaerobic cultures. These patients were divided into 4 groups based on their stage of periodontal treatment: (1) untreated patients; (2) patients after initial periodontal therapy only; (3) patients in the maintenance phase who not underwent periodontal surgery; (4) patients after periodontal surgery. RESULTS: The prevalence of mutans streptococci in the 4 study groups varied from 82% in untreated patients to 94% in maintenance patients. The mean proportion of mutans streptococci was 6.65% in maintenance patients versus 1.86% in untreated patients (p=0.005) and 2.51% in patients after scaling and root planing (p=0.041). CONCLUSIONS: The subgingival area is a microbial habitat for mutans streptococci that may be of importance in the development of root caries in periodontitis patients.

Adult↗

HAR1 mediates systemic regulation of symbiotic organ development.

Symbiotic root nodules are beneficial to leguminous host plants; however, excessive nodulation damages the host because it interferes with the distribution of nutrients in the plant. To keep a steady balance, the nodulation programme is regulated systemically in leguminous hosts. Leguminous mutants that have lost this ability display a hypernodulating phenotype. Through the use of reciprocal and self-grafting studies using Lotus japonicus hypernodulating mutants, har1 (also known as sym78), we show that the shoot genotype is responsible for the negative regulation of nodule development. A map-based cloning strategy revealed that HAR1 encodes a protein with a relative molecular mass of 108,000, which contains 21 leucine-rich repeats, a single transmembrane domain and serine/threonine kinase domains. The har1 mutant phenotype was rescued by transfection of the HAR1 gene. In a comparison of Arabidopsis receptor-like kinases, HAR1 showed the highest level of similarity with CLAVATA1 (CLV1). CLV1 negatively regulates formation of the shoot and floral meristems through cell-cell communication involving the CLV3 peptide. Identification of hypernodulation genes thus indicates that genes in leguminous plants bearing a close resemblance to CLV1 regulate nodule development systemically, by means of organ-organ communication.

Arabidopsis Proteins↗

Do root lesions tend to develop in the same people who develop coronal lesions?

OBJECTIVES: The three purposes of this study are to: (1) describe the relationship between the prevalence of coronal caries and root caries; (2) describe the relationship between the three-year incidence of coronal caries and root caries; and (3) if the two conditions are associated, develop a multiple regression model that identifies characteristics distinguishing people who had increments of both root caries and coronal caries from people who had increments of either coronal caries or root caries, or who had no new caries. METHODS: Dental examinations and interviews were conducted in the homes of a randomly selected, stratified sample of people over the age of 65 years in five North Carolina counties. The relationships between coronal and root D and DF were analyzed through contingency table analyses, and ordinal logistic regression was used to identify characteristics that differentiated people who had both coronal and root D over the three years from people who had either coronal or root D and people who had no new disease. RESULTS: Evidence of root and coronal caries in whites was much more likely to be in the form of fillings, while for blacks, it was more likely to be in the form of untreated decay. Prevalence rates of coronal and root D and DF were significantly associated for both blacks and whites. Incidence rates based on DF indicated that root and coronal caries were not associated in whites, but were associated in blacks. People more likely to experience both types of caries had more gingival recession at baseline, greater average attachment loss over the three years, and lactobacilli at baseline. In addition, the presence of Porphymonas gingivalis at three years was important for whites. CONCLUSIONS: It appears that coronal and root caries do tend to appear together in the same individuals, but fillings attenuate that relationship. The impact of dental treatment on the epidemiology of dental caries appears to be considerable and calls into question whether the F component of the caries index is related to disease as defined by epidemiologic criteria.

Aged↗