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Root deformation during root-end preparation.

Ultrasonic root-end preparation techniques have recently been introduced and revolutionized the field of endodontic surgery. However, several reports claimed that there was an increasing incidence of crack formation after ultrasonic root-end preparation. As yet, little work has focused on the root deformation during root-end preparation. Thus, the purpose of this investigation was to measure the amount of root deformation during root-end preparation with the use of microhandpiece and ultrasonic systems by using strain gauge methods, and simultaneously to detect any cracks with the aid of the stereomicroscope, stain, and an image processing system. The results demonstrated the ultrasonic instrumentation produced significantly greater strain on average than that generated with the microhandpiece system. From the viewpoint of fracture, any technique that could diminish the strain on the root would decrease the likelihood of fracture; however, no crack was observed on any resected surface of roots in this study.

Dental Instruments↗

A comparison of root fracture resistance using two root canal sealers.

AIM: The aim of this study was to compare in vitro root fracture resistance following root canal filling with either Ketac-Endo or Roth's root canal sealer. METHODOLOGY: The roots of 40 teeth with single canals were instrumented and placed randomly into four groups of 10 teeth. The prepared canals were obturated with lateral compaction using gutta-percha and Roth's 801 sealer, lateral compaction and Ketac-Endo sealer or with a single cone and Ketac-Endo sealer; the fourth group acted as unfilled controls. The roots were stored for 2 weeks in 100% humidity prior to being mounted in acrylic resin blocks. A steel tipped rod attached to an Instron testing machine was positioned against the canal opening and a slowly increasing force was applied until root fracture occurred. The results were subjected to statistical analysis using Kruskal-Wallis one-way ANOVA. RESULTS: There were no significant differences between the groups in terms of force required to vertically fracture the roots. CONCLUSIONS: Under the conditions of this study, the use of Ketac-Endo sealer in conjunction with lateral compaction or single-cone obturation techniques does not increase the fracture resistance of root-filled teeth.

Analysis of Variance↗

Studies of dental root surface caries. 2: The role of cementum in root surface caries.

Artificial caries lesions were produced in roots of teeth using an acetate buffer system, when the layer of cementum was either normal in thickness, excessively thickened by hypercementosis, or had been removed completely. The rates of lesion progression were measured in each case using polarized light microscopy to measure lesion depth. Analysis of calcium (Ca) and phosphorus (P) loss during the demineralizing process was carried out. The removal of cementum was found to significantly increase the initial rate of penetration of the lesion into the root, although this rate progressively reduced to a level consistent with that found in normal roots after seven days of demineralization. The overall depth remained consistently greater than that observed in normal roots, or when lesions were produced entirely within hyperplastic cementum. Chemical analysis also showed removal of cementum resulted in an initial doubling of the Ca and P lost from the root surface. Prior direct exposure of segments of normal roots to the oral environment was found not to significantly alter the rate of artificial lesion progression, in comparison with that in the originally protected segment of the root surface. It was concluded that an intact cementum layer has the intrinsic ability to protect the underlying dentine of exposed tooth roots against acidic demineralization and that prior exposure to the oral environment does not significantly alter this ability.

Calcium↗

Domestication-associated reduction of methyl salicylate in tomato root and its significance for resistance to root-knot nematode.

Methyl salicylate (MeSA) plays diverse roles in the aerial parts of plants. By contrast, its biosynthesis and function in roots remain poorly understood. Here, we investigated root MeSA biosynthesis and function in tomato. Genome-wide association studies (GWAS) were performed using root MeSA levels as the phenotype in a diversity panel of 167 accessions to identify associated loci. Candidate genes were biochemically characterized, and the role of MeSA in defense against root-knot nematode (RKN, Meloidogyne incognita) was evaluated using transgenic plants. MeSA was identified as a major root volatile in tomato and showed a domestication-associated reduction. GWAS revealed multiple loci associated with natural variation in root MeSA, including a major locus on Chromosome 9 encoding the salicylic acid methyltransferase (SlSAMT). SlSAMT-overexpressing plants showed reduced resistance to RKNs, whereas SlSAMT-knockdown plants exhibited enhanced resistance. Our results suggest complex roles of MeSA and the salicylic acid (SA) signaling pathway in belowground plant defense. The SA signaling pathway likely plays critical roles in protecting roots against diverse natural enemies, including RKNs. Nevertheless, RKNs appear to have co-opted MeSA as a host-location signal, and the domestication-associated reduction of root MeSA in tomato has likely contributed to enhanced resistance against RKNs.

Solanum lycopersicum↗

Inhibitory role of root hairs on transport within root culture bioreactors.

An experimental system was developed to produce root cultures of Hyoscyamus muticus with and without the profuse root hairs. Growth in the presence of 7.6 microM pyrene butyric acid (PBA) and 2.2 mM phosphate virtually eliminated root hairs, whereas growth rate, general morphology and nutrient yields remained unchanged in well-mixed flask culture. These root cultures were used to demonstrate decreased flow resistance in a tubular reactor as a result of root hair removal. To assess the impact on bioreactor performance, hairy and hairless root cultures were grown in a highly characterized 15-L bubble column bioreactor. In the absence of root hairs, the mixing was greatly enhanced; mixing times became shorter for the hairless culture at roughly 100 g (fresh weight)/L. By the end of the 3-week culture period, the mixing time of the hairy culture was 29 times longer than that of the hairless culture. The growth rate of the hairless culture in the bioreactor was as much as 2.4 times greater than growth of the hairy culture under the same conditions. The improved reactor performance was reflected in greater biomass accumulation and respiratory activity. These results show that the root hairs-which facilitate nutrient uptake in a static soil environment-are detrimental to growth in a liquid environment as an effect of their stagnating fluid flow and limiting oxygen availability.

Biological Transport↗

In vivo effects of an Er:YAG laser, an ultrasonic system and scaling and root planing on the biocompatibility of periodontally diseased root surfaces in cultures of human PDL fibroblasts.

BACKGROUND AND OBJECTIVES: The aim of the present study was to investigate the in vivo effects of an Er:YAG laser (ERL), an ultrasonic system and scaling and root planing (SRP) on the biocompatibility of periodontally diseased root surfaces in cultures of human periodontal ligament fibroblasts (PDL). STUDY DESIGN/MATERIALS AND METHODS: Forty single rooted teeth, considered for extraction due to severe periodontal destruction, have been randomly assigned to the following groups: (1) ERL at 160 mJ/pulse and 10 Hz, or (2) Vector ultrasonic system (VUS), or (3) SRP using hand instruments, or (4) untreated control (C). Immediately after instrumentation, all test and control teeth were extracted and root specimens (4 mm2) were prepared from all mesial and distal surfaces (n=80). Following the prescribed treatments, the root specimens were incubated with human PDL fibroblast cultures. Adherent cells were stained with methylene blue and counted using a reflected light microscope and the cell density per mm2 was calculated. Additionally, the cell morphology was investigated using SEM (n=8 teeth). RESULTS: Cell counts within each group yielded the following means and standard deviations (cells/mm2): ERL, 111+/-27; VUS, 75+/-25; SRP, 41+/-17; control, 25+/-11. Analysis of variance (ANOVA) revealed significant differences in the number of attached cells between the test and control groups (P<0.001, P<0.001, P<0.01, respectively). ERL and VUS treated specimens showed significantly higher numbers of cells/mm2 than the SRP group (P<0.001, respectively). The difference between the ERL and VUS group was statistically significant (P<0.001). CONCLUSIONS: The results of the present study indicate that (i) ERL, VUS, and SRP promote the attachment of PDL fibroblasts on previously diseased root surfaces, (ii) periodontally diseased root surfaces inhibit the adherence of PDL fibroblasts, and (iii) the surface structure of ERL and VUS instrumented roots seem to offer better conditions for the adherence of PDL fibroblasts than SRP.

Aged↗

Evaluation of methods to measure differential 15N labeling of soil and root N pools for studies of root exudation.

To study patterns of root exudation, the effectiveness of different techniques for in situ 15N labeling of Brassica napus, Centaurea jacea and Lolium perenne with ammonium nitrate was tested. Stem infiltration was found to effectively label plants with thicker stems, whereas, for grass species, cutting and immersing the leaf tips into 15N solution proved to be most effective. A microdiffusion technique to isolate ammonium, combined with conventional cation-exchange chromatography to separate nitrate from amino-N compounds thereafter, was found suitable for separation of the N fractions of plant and soil extracts for 15N determination. All three species were then cultivated in nutrient solution and labeled with 15NH4 15NO3 by stem feeding for 42 hours. Kinetics of 15N labeling of bulk roots and shoots as well as hot water extractable material were assessed, and up to 1.1 at% 15N excess (APE) was found in nutrient solutions. The main amino acids exuded by L. perenne were glycine, serine, alanine and aspartic acid. To assess the suitability of this set of methods to study root exudation in field settings, L. perenne was grown without fertiliser addition in pots containing low-nutrient soil. Plants were 15N labeled via tip immersion and 15N and N concentrations were analysed in shoots, roots and soils during a 48-h interval. Shoots reached 1.25 APE, roots and soil 0.10 and 0.005 APE, respectively. Between 4% (48 h) and 6% (24 h) of total plant 15N was exuded by roots into the soil. In roots amino acids comprised the largest proportion of the soluble 15N pool, whereas soil 15N levels were similar for amino acids and ammonium, exceeding those of nitrate. Mechanisms for the shift within N fractions from roots to soils are briefly discussed.

Chromatography, Ion Exchange↗

Sheaths of the spinal nerve roots. Permeability and structural characteristics of dorsal and ventral spinal nerve roots of the rat.

The present study was carried out to investigate the permeability of normal spinal nerve root sheaths around dorsal and ventral roots in the rat. In vivo studies were performed using Evans blue-albumin and lanthanum chloride as tracers. The Evans blue-albumin complex is macromolecular in size and lanthanum ions are small and easily visible in the electron microscope. Both tracers were injected into the subarachnoid space and 15 min later samples were taken and further processed for detection of tracer. Postmortem studies with lanthanum was also performed. Following fixation by cardiac perfusion with fixative without tracer, lanthanum chloride was added to the fixative and applied directly to exposed spinal cord including the spinal nerve roots. Macroscopical examination showed Evans blue staining of the superficial blood vessels of the spinal cord, but no staining of the parenchyma of either spinal cord or nerve roots. Fluorescence microscopy revealed, in addition to a bright red fluorescence of root sheaths, a faint longitudinally orientated red fluorescence in the endoneurium of the nerve roots, indicating the presence of the dye-albumin complex. In both in vivo and post-mortem lanthanum studies, the tracer was detected between cell layers of the nerve root sheath and in invaginations of the plasma membrane of these cells, as well as inside the nerve root parenchyma. Some of the cells of the sheaths in post-mortem animals were diffusely marked with intracellular tracer. The endo-radicular lanthanum was most often seen superficially, but lanthanum could occasionally be detected deeper in the parenchyma in the post mortem studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Transgenic white clover. Studies with the auxin-responsive promoter, GH3, in root gravitropism and lateral root development.

We report improved method for white clover (Trifolium repens) transformation using Agrobacterium tumefaciens. High efficiencies of transgenic plant production were achieved using cotyledons of imbibed mature seed. Transgenic plants were recovered routinely from over 50% of treated cotyledons. The bar gene and phosphinothricin selection was shown to be a more effective selection system than nptII (kanamycin selection) or aadA (spectinomycin selection). White clover was transformed with the soybean auxin responsive promoter, GH3, fused to the GUS gene (beta-glucuronidase) to study the involvement of auxin in root development. Analysis of 12 independent transgenic plants showed that the location and pattern of GUS expression was consistent but the levels of expression varied. The level of GH3:GUS expression in untreated plants was enhanced specifically by auxin-treatment but the pattern of expression was not altered. Expression of the GH3:GUS fusion was not enhanced by other phytohormones. A consistent GUS expression pattern was evident in untreated plants presumably in response to endogenous auxin or to differences in auxin sensitivity in various clover tissues. In untreated plants, the pattern of GH3:GUS expression was consistent with physiological responses which are regarded as being auxin-mediated. For the first time it is shown that localised spots of GH3:GUS activity occurred in root cortical tissue opposite the sites where lateral roots subsequently were initiated. Newly formed lateral roots grew towards and through these islands of GH3:GUS expression, implying the importance of auxin in controlling lateral root development. Similarly, it is demonstrated for the first time that gravistimulated roots developed a rapid (within 1 h) induction of GH3:GUS activity in tissues on the non-elongating side of the responding root and this induction occurred concurrently with root curvature. These transgenic plants could be useful tools in determining the physiological and biochemical changes that occur during auxin-mediated responses.

Agrobacterium tumefaciens↗

Transgenic Medicago truncatula plants obtained from Agrobacterium tumefaciens -transformed roots and Agrobacterium rhizogenes-transformed hairy roots.

Medicago truncatula, barrel medic, is a forage crop that has been developed into a model legume. The development of new transformation methods is important for functional genomic studies in this species. Based on Agrobacterium tumefaciens-mediated transformation of root explants, we developed an effective system for producing M. truncatula (genotype R108) transgenic plants. Among the four A. tumefaciens strains (AGL1, C58C1, EHA105 and LBA4404) tested, EHA105 and AGL1 were most effective in regenerating transgenics. Callus induction frequency from root explants was 69.8%, and plantlet/shoot regeneration frequency was 41.3% when EHA105 was used. Transgenic nature of the regenerated plants was confirmed by PCR and Southern hybridization analyses. Progeny analysis revealed stable Mendelian meiotic transmission of transgenes. Because M. truncatula is particularly useful for the study of root endosymbiotic associations, we further developed a plant regeneration system from A. rhizogenes-transformed hairy roots of M. truncatula. Fertile true transgenic plants were regenerated from the hairy roots, thus allowing the assessment of gene functions at the whole plant level. Segregation analysis revealed that the hairy root genes could be segregated out in the progenies. By coupling A. rhizogenes-mediated hairy root transformation and the regeneration system reported here, once potential genes of interest are identified, the transformed hairy roots carrying such genes could be directly regenerated into plants for more detailed characterization of the genes.

Acetyltransferases↗

Studies on peri-root tissue formation around new type artificial root made of dense hydroxyapatite.

The aim of this study was to examine the wound healing process and tissue development around a new type of artificial root made of dense hydroxyapatite. The newly tailored artificial roots, which have characteristic corrugated configurations with smooth root surfaces, were implanted immediately after extraction of the premolars of adult dogs. After preliminary experiments, the following studies were carried out: for the observation of tissue formation around the artificial root, artificial root with surrounding tissue were extirpated after fixed periods (6-32 weeks) of implantation; to study tissue formation at the artificial root surface, undecalcified specimens with polished surfaces for microanalysis and decalcified specimens for light microscopic observation were prepared after 72 weeks of implantation. The following results were obtained: around the artificial roots in the jawbone, fibrous tissue formation with angled orientation, bone formation resembling alveolar bone proper, and calcified substance formation attaching to the root surface were observed.

Animals↗

Vertical root fracture and root distortion: effect of spreader design.

The incidence of vertical root fractures and the amount of root distortion created during lateral condensation of gutta-percha with either D11 spreaders or B-finger pluggers were evaluated in vitro. Fifty-five extracted human, single-rooted teeth were instrumented using the step-back flare technique. Ten teeth served as positive controls (obturation to the point of fracture) and five teeth as negative controls (prepared but not obturated). Strain gauges were attached to the root surfaces. In the experimental group, 20 teeth were obturated using a D11 spreader and 20 with a B-finger plugger. Recordings were made of root distortion (expansion) created during obturation. Then, after sectioning the teeth, root surfaces of obturated samples were examined for fractures under the scanning electron microscope. Only the more tapered spreader, the D11, produced vertical root fractures, although very few in number. Also, the D11 spreader caused greater root distortion than did the B-finger plugger.

Dental Stress Analysis↗

Identification of genes enriched in rice roots of the local nitrate treatment and their expression patterns in split-root treatment.

To identify nitrate-induced genes in rice roots, a nitrate-induced root subtracted library was constructed using the suppression subtractive hybridization (SSH) method and a split-root experiment. Based on forward and reverse screening, 37 known genes and 55 novel genes were identified to be up-regulated in nitrate-supplied side roots. The known genes are involved in nitrogen (N) uptake and assimilation, sugar transport and organic acid metabolism, signal transduction, protein synthesis and degradation, plant resistance, hormone metabolism and cell division. Most of these genes were stronger up-regulated in nitrate-supplied than in nitrate-deficient side roots, which suggests that nitrate may regulate expression of these genes. Expression pattern analysis also revealed that a fast cycle of assimilated N metabolites occurred between root and shoot. Transcriptions of some nitrate-induced genes were also up-regulated in nitrate-deficient side roots, which indicates that nitrate and/or in coordinating with N metabolites from shoots, regulate the transcription of many genes. The enhancement of carbon (C) source partition, the expressions of genes for signal transduction and transcription regulation, auxin transport and ethylene synthesis, and cyclin-dependent kinases may be responsible for the stimulated lateral root growth by nitrate.

Adaptation, Physiological↗

Host-root exudates increase gene expression of asparagine synthetase in the roots of a hemiparasitic plant Triphysaria versicolor (Scrophulariaceae).

Triphysaria is a facultative root parasite in the Scrophulariaceae family. Similar to other related parasites, the development of the parasitic life cycle is initiated by molecular signals released from appropriate host roots. Using a differential display, we isolated cDNAs preferentially abundant in T. versicolor roots exposed to Trifolium repens (white clover) root exudates in vitro. Sequence analysis indicated that one of the differentially expressed cDNAs had significant homology to the nitrogen-assimilating enzyme, asparagine synthetase (AS). T. versicolor AS cDNA clones were isolated and placed into three distinct classes on the basis of nucleotide sequence variations. All three classes encoded identical AS proteins. AS was expressed in both roots and shoots of in-vitro-cultured T. versicolor. Steady-state levels of AS mRNA increased in T. versicolor roots several-fold when seedlings were exposed to exudate obtained from hydroponically grown Arabidopsis thaliana roots. Therefore, AS transcript levels increased in response to exudates from two different hosts (Trifolium and Arabidopsis). The T. versicolor AS message levels increased to a similar magnitude when seedlings were incubated in the dark. Interestingly, AS levels were unaffected by treatment with the Striga haustoria inducer 2,6-dimethoxybenzoquinone. The potential role of AS in root parasitism is discussed.

Amino Acid Sequence↗

Effects of Astragali root and Hedysari root on the murine B and T cell differentiation.

It has traditionally been known in Asian countries that the Astragali and Hedysari roots have similar beneficial effects. They have been applied in clinical practice for immunological diseases, we investigated their effects on specific antibody production, B cells and T cells in BALB/c mice immunized with ovalbumin (OVA). Both the Astragali and Hedysari roots reduced the production of IgG2a in the primary response to OVA. Hedysari root reduced the production of IgG2a in the secondary response. Both reduced the production of IgM, suppressed IL-6 production in spleen cells. It was presumed that the suppression of IgG2a production was induced by the suppression of IL-6, which is an antibody production inducer. It was found that the Hedysari root increased the B cells and activated them, and the Astragali root also tended to increase them. In addition, Hedysari root decreased the proportions of IFN-gamma-producing cells in splenic CD4(+) T lymphocytes. It was shown that the Astragali and Hedysari roots could act as a growth factor of B cells by increasing the proportion of the total B cells and activated B cells. In conclusion, both the Astragali and Hedysari roots showed the same actions, but the latter was more effective.

Animals↗

Anti-hepatotoxic effects of root and root callus extracts of Cichorium intybus L.

The natural root and root callus extracts of Cichorium intybus were compared for their anti-hepatotoxic effects in Wistar strain of Albino rats against carbon tetrachloride induced hepatic damage. The increased levels of serum enymes (aspartate transaminase, alanine transaminase) and bilirubin observed in rats treated with carbon tetrachloride were very much reduced in the animals treated with natural root and root callus extracts and carbon tetrachloride. The decreased levels of albumin and proteins observed in rats after treatment with carbon tetrachloride were found to increase in rats treated with natural root and root callus extracts and carbon tetrachloride. These biochemical observations were supplemented by histopathological examination of liver sections. Results of this study revealed that Cichorium intybus root callus extract could afford a better protection against carbon tetrachloride induced heptocellular damage as compared to the natural root extract.

Alanine Transaminase↗

Further root colonization by arbuscular mycorrhizal fungi in already mycorrhizal plants is suppressed after a critical level of root colonization.

An established arbuscular mycorrhizal symbiosis suppresses further mycorrhization. It is not clear whether the observed suppressional effect is linked with the level of root colonization or not. In the present work we studied the effect of the degree of root colonization by the arbuscular mycorrhizal fungus Glomus mosseae on further root colonization by G. mosseae. At different time points barley plants grown in split-root compartments were pre-inoculated on one half of the split-root system with G. mosseae. Sequential inoculation resulted in different colonization levels. Thereafter, the second half of the split root system was inoculated. The results indicate an enhanced suppression of root colonization on the second side of the split-root system when colonization levels increased on the first side.

Colony Count, Microbial↗

A trans-zeatin riboside in root xylem sap negatively regulates adventitious root formation on cucumber hypocotyls.

Shoot cultures of cucumber were used to analyse the roles of root-derived substances in adventitious root formation on hypocotyl tissues. Xylem sap collected from the roots of squash had a strong inhibitory effect on the formation of hypocotyl adventitious roots. Double-solvent extraction followed by fractionation with both normal and reverse phase column chromatographies and analysis by liquid chromatography/tandem mass spectrometry identified trans-zeatin riboside (ZR) as the primary suppressor of adventitious root formation. ZR was the predominant cytokinin present in the xylem sap, occurring at a concentration of 2x10(-8 )M. Application of ZR at concentrations from 3.16x10(-9) M effected inhibition of adventitious root formation. These results suggest that ZR transported from roots via xylem sap may act as an endogenous suppressor of hypocotyl adventitious root formation in planta.

Adenosine↗