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The maize Dwarf3 gene encodes a cytochrome P450-mediated early step in Gibberellin biosynthesis.

Gibberellins (GAs) are phytohormones required for normal growth and development in higher plants. The Dwarf3 (D3) gene of maize encodes an early step in the GA biosynthesis pathway. We transposon-tagged the D3 gene using Robertson's Mutator (Mu) and showed that the mutant allele d3.2::Mu8 is linked to a Mu8 element. The DNA flanking the Mu8 element was cloned and shown to be linked to the d3 locus by mapping in a high-resolution population developed by selecting for recombination between d3 and linked genetic markers. To establish unambiguously the identity of the cloned gene as D3, a second mutant allele of D3 (d3.4) was also cloned and characterized using the d3.2::Mu8 sequences as a probe. d3.4 was found to have a novel insertion element, named Sleepy, inserted into an exon. A third mutant allele, d3.1, which has the same size 3' restriction fragments as d3.4 but different 5' restriction fragments, was found to contain a Sleepy insertion at the same position as d3.4. On the basis of the pedigree, Sleepy insertion, and restriction map, d3.1 appears to represent a recombinational derivative of d3.4. The D3 gene encodes a predicted protein with significant sequence similarity to cytochrome P450 enzymes. Analysis of D3 mRNA showed that the D3 transcript is expressed in roots, developing leaves, the vegetative meristem, and suspension culture cells. We detected reduced D3 mRNA levels in the mutant allele d3.5.

Alleles↗

Two sweetpotato ADP-glucose pyrophosphorylase isoforms are regulated antagonistically in response to sucrose content in storage roots.

The transcriptional regulation of ADP-glucose pyrophosphorylase (AGPase) genes in detached leaves in response to exogenous sucrose has been investigated earlier; however the effects of endogenous sucrose on AGPase gene transcription in leaves or starch-accumulating tissues have not yet been determined. We therefore have investigated the relationship between endogenous sucrose content in the storage tissues of sweetpotato (Ipomoea batatas cv. Yulmi) and the rate of transcription of the two sweetpotato AGPase isoforms, ibAGP1 and ibAGP2, by means of transient expression analysis of their promoters. Sequence analysis of the two promoters identified putative sucrose-responsive elements on the ibAGP1 promoter and, conversely, putative sucrose-starvation elements on the ibAGP2 promoter. Transient expression analyses on transverse storage root sections revealed that the ibAGP1 and ibAGP2 promoters directed strong expression in the sweetpotato storage roots (diameter: 1.5 cm). Sucrose contents of the sweetpotato storage roots were positively correlated with growth of the storage root. In the storage roots, ibAGP1 promoter activity became stronger with increasing endogenous sucrose levels, while ibAGP2 promoter activity became markedly weaker. Consequently, ibAGP2 was expressed primarily during the early stages of storage root development, whereas ibAGP1 was abundantly expressed in the later stages, during which a profound level of starch accumulation occurs. The antagonistic regulation of the two promoters in response to endogenous sucrose levels was also confirmed in carrot (Daucus carota L. cv. Hapa-ochon) taproots.

Base Sequence↗

[Results after short- and long-term observations of bone transplantation to vertical, marginal bone defects].

The effect of autologue bone transplantation to 26 vertical, marginal bone defects has been investigated on a long-term scale ranging from 2 to 26 years. The operation technique included excision of gingival pocket epithelium, removal of granulation tissue, and cleansing of the root surface with scalers. The bone cavities were filled preferably with cancellous bone chips, usually from the patients' upper third molar region. After suturing, the area was covered with a wound paste for 12-14 days. Loosened teeth were splinted for at least 2 months. In 25 of 26 teeth (96%) a bone regeneration was observed radiographically. The maximum healing of 22 cases occurred 2-12 months (median 6 months) postoperatively (Table 1). The maximum healing was maintained for more than half of the teeth (56%) at the last control, median 16 years (range 2-26 years), and none of these teeth were extracted of periodontal causes. They might therefore be expected to function for still a long time. Eight teeth (32%) showed more or less recurrence when controlled 5 to 15 years (median 10 years) after operation and 6 of the 8 teeth with recurrence were extracted. Three teeth (12%) developed root resorption and were extracted 6, 10 and 14 years postoperatively. As most of the complications occurred after extended observation periods, this factor has to be taken into account when prognosis of periodontal treatment procedure is considered.

Adult↗

The Arabidopsis trehalose-6-P synthase AtTPS1 gene is a regulator of glucose, abscisic acid, and stress signaling.

In Arabidopsis (Arabidopsis thaliana), trehalose is present at almost undetectable levels, excluding its role as an osmoprotectant. Here, we report that overexpression of AtTPS1 in Arabidopsis using the 35S promoter led to a small increase in trehalose and trehalose-6-P levels. In spite of this, transgenic plants displayed a dehydration tolerance phenotype without any visible morphological alterations, except for delayed flowering. Moreover, seedlings overexpressing AtTPS1 exhibited glucose (Glc)- and abscisic acid (ABA)-insensitive phenotypes. Transgenic seedlings germinated on Glc were visibly larger with green well-expanded cotyledonary leaves and fully developed roots, in contrast with wild-type seedlings showing growth retardation and absence of photosynthetic tissue. An ABA dose-response experiment revealed a higher germination rate for transgenic plants overexpressing AtTPS1 showing insensitive germination kinetics at 2.5 mum ABA. Interestingly, germination in the presence of Glc did not trigger an increase in ABA content in plants overexpressing AtTPS1. Expression analysis by quantitative reverse transcription-PCR in transgenic plants showed up-regulation of the ABI4 and CAB1 genes. In the presence of Glc, CAB1 expression remained high, whereas ABI4, HXK1, and ApL3 levels were down-regulated in the AtTPS1-overexpressing lines. Analysis of AtTPS1 expression in HXK1-antisense or HXK1-sense transgenic lines suggests the possible involvement of AtTPS1 in the hexokinase-dependent Glc-signaling pathway. These data strongly suggest that AtTPS1 has a pivotal role in the regulation of Glc and ABA signaling during vegetative development.

Abscisic Acid↗

Immunocytochemical investigation of the rat cementoblast phenotype.

Recent studies have suggested that cementoblasts may be derived from osteoblast progenitor cells, although the cementoblast phenotype has not been extensively characterized. This immunocytochemical study was carried out to investigate the expression by rat cementoblasts of a number of proteins which are characteristic of the osteoblast phenotype. Paraffin sections from developing rat tooth germs and from fully formed adult rat teeth with surrounding tissues, were incubated with antibodies to type I & III collagen, osteocalcin, transforming growth factor beta (TGE beta), and insulin-like growth factor 1 (IGF1). Frozen sections and unfixed resin-embedded sections were stained for alkaline phosphatase activity. Cementum and bone matrix were strongly positive for type I collagen, although there was only weak staining for type III collagen. Cementum was also positive for osteocalcin, which was particularly strong in the matrix of acellular cementum. Most osteoblasts and cementoblasts of the cellular cementum showed intense staining for TGF beta and IGF1, although some cementocytes and osteocytes were negatively stained. The osteoblast- specific anti-E11 mAb reacted strongly with cementoblasts and newly formed cementocytes in the cellular cementum. Cells associated with acellular cementum did not express TGF beta, IGF1 or stain positively with anti-E11 antibody at any time during root development. Cementoblasts were weakly or negatively stained for alkaline phosphatase in contrast to the osteoblasts examined, which may reflect the low level of synthetic activity in cementoblasts. These results demonstrate that osteoblasts and cementoblasts of cellular cementum share many phenotypic characteristics, and also suggest that there may be phenotypic differences between cementoblasts associated with cellular and acellular cementum.

Animals↗

[Epithelial cell rests of Malassez: tissue, cell, and molecular biology].

This review presents an analysis of the current data on the structural and functional organization and biological significance of epithelial cell rests of Malassez (ERM), which are formed by persisting fragments of epithelial (Hertwig) sheath that participates in the tooth root development. ERM, found within the periodontal ligament, undergo intensive age-related involution and are traditionally described as small clusters of functionally inactive cells. Meanwhile, recent findings are indicative of high functional activity of ERM which are both producers of and targets for various cytokines, growth factors, adhesive substances, their receptors and other biologically active molecules. It is suggested that ERM participate in sustaining the optimal conditions for the normal functioning and regeneration of periodontium structural components. When periodontal tissue homeostasis is disturbed, ERM may grow up and acquire the capacity to destroy the surrounding tissues.

Cytokines↗

Liberation of amino acids by heterotrophic nitrogen fixing bacteria.

Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed.

Amino Acids↗

Arabidopsis SHY2/IAA3 inhibits auxin-regulated gene expression.

In Arabidopsis, SHY2 encodes IAA3, a member of the auxin-induced Aux/IAA family. Gain-of-function mutations in SHY2/IAA3 cause enlarged cotyledons, short hypocotyls, and altered auxin-regulated root development. Here we show that the gain-of-function mutation shy2-2 decreases both the induction and repression of auxin-regulated genes, suggesting that SHY2/IAA3 acts as a negative regulator in auxin signaling. shy2-2 affects auxin induction of many previously characterized primary response genes, implying that it might repress primary auxin responses. In addition, shy2-2 also affects expression of multiple auxin-nonresponsive genes. Light regulates expression of SHY2/IAA3, suggesting a possible link between light and auxin response pathways.

Arabidopsis↗

Arabidopsis SGT1b is required for SCF(TIR1)-mediated auxin response.

The SCF(TIR1) complex is a central regulator of the auxin response pathway in Arabidopsis. This complex functions as a ubiquitin protein ligase that targets members of the auxin/indoleacetic acid (Aux/IAA) family of transcriptional regulators for ubiquitin-mediated degradation in response to auxin. In an attempt to identify additional factors required for SCF(TIR1) activity, we conducted a genetic screen to isolate enhancers of the auxin response defect conferred by the tir1-1 mutation. Here, we report the identification and characterization of the eta3 mutant. The eta3 mutation interacts synergistically with tir1-1 to strongly enhance all aspects of the tir1 mutant phenotype, including auxin inhibition of root growth, lateral root development, hypocotyl elongation at high temperature, and apical dominance. We isolated the ETA3 gene using a map-based cloning strategy and determined that ETA3 encodes SGT1b. SGT1b was identified recently as a factor involved in plant disease resistance signaling, and SGT1 from barley and tobacco extracts was shown to interact with SCF ubiquitin ligases. We conclude that ETA3/SGT1b is required for the SCF(TIR1)-mediated degradation of Aux/IAA proteins.

Alleles↗

[Identification of developmental stages of wisdom teeth on orthopantomograms of adolescents and young adults as an aid for forensic-odontological age-estimations: predictive values for the chronological age of 18 years].

UNLABELLED: Dental findings are an important source of information regarding forensic-odontologic determination of identity and age. A main part of dental age assessment is the determination of the wisdom teeth' development. The aim of this study was to answer the question whether the development of wisdom teeth can give information for the task of individual age estimation. Thereby, the investigation could contribute to age estimation efforts that are carried out on persons with an unknown age and where it is of high importance to know as exactly as possible the age of a person, e.g. the age of 18 years or older, for determining the punishment for a criminal offence. MATERIAL AND METHODS: The orthopantomograms of 1053 outpatients were evaluated. The patients were aged 14 to 24 years. The developmental stages of the wisdom teeth were determined according to Demirjian. The data were calculated using statistical tools, like descriptive and inferential statistics in order to provide predictive values for the age to be above or below 18 years. RESULTS: The correlation between the developmental stages of wisdom teeth and the chronological age was high in general. Some values allowed the prediction of an age above 18 years with a probability of more than 70% to 80%, in some instances even more than 90%. The most appropriate developmental stage to determine the age of 18 years was A 1/2 (completed root development, apex open). The positive predictive value of this finding was 94%. DISCUSSION: This study provides for the first time predictive values for the estimation whether or not a person is 18 years of age, based on the evaluation of developmental stages of wisdom teeth on orthopantomograms only. These results should be compared to data from other populations gained by the same method.

Adolescent↗

Multiple functions for NGF receptor in developing, aging and injured rat teeth are suggested by epithelial, mesenchymal and neural immunoreactivity.

We have used immunocytochemistry to analyse expression of nerve growth factor receptor (NGFR) in developing, aging and injured molar teeth of rats. The patterns of NGFR immunoreactivity (IR) in developing epithelia and mesenchyme matched the location of NGFR mRNA assayed by in situ hybridization with a complementary S35-labeled RNA probe. The following categories of NGFR expression were found. (1) There was NGFR-IR in the dental lamina epithelium and in adjacent mesenchyme during early stages of third molar formation. (2) NGFR-IR nerve fibers were posterior and close to the bud epithelium. (3) During crown morphogenesis NGFR expression was prominent in internal enamel epithelium and preodontoblasts; it faded as preameloblasts elongated and as odontoblasts began to make predentin matrix; and it was weak or absent from outer enamel epithelium, the cervical loop, and differentiated ameloblasts and odontoblasts. (4) When NGFR-IR nerve fibers entered the molars late in the bell stage, they innervated the most mature peripheral pulp and dentin in an asymmetric pattern which correlated more with asymmetric enamel synthesis than with mesenchymal NGFR-IR distribution. (5) The mesenchymal pulp cells continued to have intense NGFR expression in adult teeth, especially near coronal tubular dentin. (6) The pulpal NGFR-IR decreased in very old rats or subjacent to reparative dentin (naturally occurring or experimentally induced). (7) During root formation, the preodontoblasts had NGFR-IR but most root mesenchymal cells and Hertwig's epithelial root sheath did not. This work suggests that there are important epithelial and mesenchymal targets of NGF regulation during molar morphogenesis that differ for crown and root development and that do not correlate with neural development. The continuing expression of NGFR-IR by pulpal mesenchymal cells in adult rats was most intense near coronal odontoblasts making tubular dentin; and it was lost during aging, or subjacent to sites of dentin injury that caused a phenotypic change in the odontoblast layer.

Aging↗

HAT3.1, a novel Arabidopsis homeodomain protein containing a conserved cysteine-rich region.

Homeodomain proteins have been shown to play a major role in the development of various organisms. A novel Arabidopsis homeodomain protein has been isolated based on its capability to interact with a DNA motif derived from the light-induced cab-E promoter of Nicotiana plumbaginifolia. The homeodomain of this protein, designated HAT3.1, differs substantially from those in other plant homeobox proteins identified so far. Furthermore, HAT3.1 is unique among other Arabidopsis proteins in that it does not contain a leucine zipper motif following the homeodomain. HAT3.1 is further characterized by an N-terminal region that shares substantial sequence similarity with the maize homeodomain protein Zmhox1a. Within this conserved region, the presence of eight regularly spaced cysteine/histidine residues was observed reminiscent of other metal-binding domains. Based on the strong evolutionary conservation of this domain, it is proposed that this region represents a novel protein-motif which is denoted PHD-finger (plant homeodomain-finger). In vitro DNA binding studies demonstrated that HAT3.1 is capable of interacting with any DNA fragment larger than 100 bp. Interestingly, a deletion of the N-terminal PHD-finger domain completely abolished DNA binding, suggesting that this region may play an important functional role in protein-protein or protein-DNA interaction. HAT3.1 mRNA was primarily detected in root tissue, implying a regulatory function of this protein in root development.

Amino Acid Sequence↗

The etiology and prevalence of gingival recession.

BACKGROUND: Gingival recession in its localized or generalized form is an undesirable condition resulting in root exposure. The result often is not esthetic and may lead to sensitivity and root caries. Exposed root surfaces also are prone to abrasion. The purpose of this article is to describe the prevalence, etiology and factors associated with gingival recession. TYPES OF STUDIES REVIEWED: The authors reviewed cross-sectional epidemiologic studies of gingival recession and found that they correlated the prevalence of recession to trauma, sex, malpositioned teeth, inflammation and tobacco consumption. The recent surveys they reviewed revealed that 88 percent of people 65 years of age and older and 50 percent of people 18 to 64 years of age have one or more sites with recession. The presence and extent of gingival recession also increased with age. RESULTS: More than 50 percent of the population has one or more sites with gingival recession of 1 mm or more. The prevalence of gingival recession was found in patients with both good and poor oral hygiene. It has been proposed that recession is multifactorial, with one type being associated with anatomical factors and another type with physiological or pathological factors. Recession has been found more frequently on buccal surfaces than on other aspects of the teeth. CLINICAL IMPLICATIONS: Dentists should be knowledgeable about the etiology, prevalence and associating factors of gingival recession, as well as treatment options, so that appropriate treatment modalities can be offered to patients. Treatments for gingival recession include gingival grafting, guided tissue regeneration and orthodontic therapy. Such treatments typically result in esthetic improvement, elimination of sensitivity and a decreased risk of developing root caries.

Adolescent↗

Cytokine secretion of periodontal ligament fibroblasts derived from human deciduous teeth: effect of mechanical stress on the secretion of transforming growth factor-beta 1 and macrophage colony stimulating factor.

The periodontal ligament may play an important role in tooth eruption, root development and resorption. The tissue physiologically receives mechanical force during mastication. We focused on the effects of intermittent mechanical strain on the cytokine synthesis of periodontal ligament (PDL) fibroblasts in vitro. The cells were derived from human periodontal ligament of deciduous teeth (HPLF-Y) and permanent teeth (HPLF). The two kinds of PDL cells and human gingival fibroblasts (HGF) were cultured in flexible bottomed culture plates. The cells were mechanically stretched at 5% elongation, 3-cycles/min for 24 h on d 7 in culture using a Flexercell strain unit. After the stretching, we measured DNA content and alkaline phosphatase activity in the cell layer, transforming growth factor beta 1 (TGF-beta 1) and macrophage colony stimulating factor (M-CSF) contents in the conditioned medium. The TGF-beta 1 level in the conditioned medium of HPLF was significantly higher than that of HPLF-Y and HGF. It was stimulated by mechanical stretching only on HPLF, whereas no significant effect was observed on HPLF-Y and HGF. M-CSF secretion was inhibited by the stretching on all of HPLF, HPLF-Y and HGF. 1 alpha, 25 dihydroxy vitamin D3 (D3) stimulated M-CSF secretion into the culture medium of both HPLF and HPLF-Y, but the stretching inhibited M-CSF secretion and completely blocked the enhancement by D3. These data suggest that periodontal ligament cells synthesize and secrete the molecules as autocrine or paracrine factors that affect bone remodelling and root resorption and the level of those factors change in response to mechanical stress.

Alkaline Phosphatase↗

Light inactivation of Arabidopsis photomorphogenic repressor COP1 involves a cell-specific regulation of its nucleocytoplasmic partitioning.

Arabidopsis COP1 acts as a repressor of photomorphogenesis in darkness, and light stimuli abrogate this suppressive action. COP1, when fused to beta-glucuronidase (GUS), is enriched in the nucleus in darkness, but not in the light, in hypocotyl cells of Arabidopsis seedlings and epidermal cells of onion bulbs. In Arabidopsis hypocotyl cells, the nuclear GUS-COP1 level changes in response to dark-light transitions and quantitatively correlates with the extent of repression of photomorphogenic development. In root cells, GUS-COP1 is constitutively nuclear, consistent with an established role of COP1 in suppressing root chloroplast development in both light and darkness. We conclude that COP1 acts inside the nucleus to suppress photomorphogenesis and that light inactivation of COP1 involves a cell type-specific control of its nucleocytoplasmic partitioning.

Allium↗

Scots pine expresses short-root-specific peroxidases during development.

Nine short-root-specific proteins from Scots pine (Pinus sylvestris L.) detected and isolated as individual spots by 2D-PAGE were identified. The similar peptide mass maps obtained for all nine polypeptide spots together with lectin-blotting results suggest that they represent forms of the same modified protein. N-Terminal sequence analysis of two of the peptides showed high similarity to peroxidases. RT-PCR with oligonucleotide primers corresponding to determined peptide sequences and conserved regions in plant peroxidases led to isolation of Psyp1 cDNA which is most abundantly expressed in short roots. Psyp1 is the first peroxidase cDNA to be isolated from the genus Pinus. It encodes a 363-amino-acid class-III peroxidase with a calculated molecular mass of 35.7 kDa and theoretical pI of 4.74. The predicted PSYP1 amino-acid sequence is grouped with other class-III peroxidases in phylogenetic analyses, but it has a unique amino-acid sequence which may be associated with its function in short roots or with its phylogenetic group. The presence of a signal sequence for extracellular transport indicates that PSYP1 belongs to the group of secreted class-III peroxidases. The presence of 10 tyrosine residues and putative auxin-binding regions in PSYP1 suggests that the function of the enzyme is associated with cell-wall formation in short roots. The downregulation of Psyp1 expression in symbiotic short roots hosting the ectomycorrhizal fungus Suillus bovinus is perhaps related to the change in cell-wall structure necessary for ectomycorrhizal development.

Amino Acid Sequence↗

An adolescent female Neandertal mandible from Montgaudier Cave, Charente, France.

In 1974, an incomplete human mandible was discovered in the site of Montgaudier Cave, along the Tardoire (Charente), France. The mandible was found in association with stone tools and animal bones in geological deposits referable to the very end of the Middle Pleistocene or the beginning of the Upper Pleistocene. The mandible preserves much of the anterior part of the body and three permanent teeth: left lateral incisor, canine and first molar. Estimates based on tooth eruption of modern humans, as well as occlusal wear and root development, suggest an age at death of between 12.5 and 14.5 years. Morphologically, the fossil possesses features, such as a lack of a chin and multiple mental foramina, which have been observed on immature Neandertal mandibular specimens from Europe. Comparison with these immature European Neandertals indicates that the jaw and teeth of the Montgaudier mandible are small for its chronological age, suggesting it was that of a female.

Adolescent↗

Cytokinin stress changes the developmental regulation of several defence-related genes in tobacco.

Tobacco shoots exposed to elevated endogenous or exogenous cytokinin levels are unable to develop roots and lack apical dominance. We have isolated cDNA copies of five mRNA species that accumulate to elevated levels in such cytokinin-stressed shoots via differential screening of a cDNA library of transgenic shoots which contain an active T-DNA cytokinin gene (T-cyt gene) from Agrobacterium tumefaciens. Four of the cDNA clones were found to correspond to plant defence-related mRNAs, encoding extensin, chitinase, PR-1 and a PR-1-like protein, respectively. In normal tobacco plants PR-1 mRNA is relatively rare in all organs. The other four mRNAs occur at relatively low levels in shoots, especially in leaves, but are very prevalent in roots. Extensin mRNA, for example, is not detectable in leaves, while it is an abundant mRNA in roots and stems. In normal shoots cultured on cytokinin-containing medium all five mRNAs accumulate to elevated levels, similar to those found in transgenic T-cyt shoots. We conclude that the imposed cytokinin stress causes changes in the tissue-specific control of the levels of several defence-related mRNA species in tobacco.

Journal Article↗