[Effect of Trichodesma incanum seeds on the development of fatty dystrophy of the liver].
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Animals↗
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[The dependence of the development of irradiated seed on the temperature of germination].
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Mustard Plant↗
Strategic planning. From the ground up: planting the seeds of network development.
It is critical to understand and accept the intense effort and planning needed for successful network development. No model exists that can be immediately implemented successfully. Every organization involved in network development will have to become familiar with the issues, benefits and risks, as well as involve the key stakeholders, which include administration, board and health professional providers from physicians through allied professionals. The outcome of this development can be a very successful, efficient new delivery model that will benefit the users, the payers and the providers. Although difficult and sometimes boring, the methodical and thorough movement through the process of developing a regional community network can greatly enhance outcome. The shared vision of the network should be exciting, and should make participants search for the clear image of what will be developed. All should work jointly as partners to accomplish that vision. Although there may be facilitation and assistance, it will be the providers and stakeholders who will be most affected by the network. It is their commitment, effort, energy and experience that will lead to success.
Comprehensive Health Care↗
Expression of thaumatin-like permatin PR-5 genes switches from the ovary wall to the aleurone in developing barley and oat seeds.
Permatins are antifungal thaumatin-like proteins (TLPs) of the PR-5 family of pathogenesis-related proteins. They occur in many cereals, but little is known of their expression and roles. Permatin cDNA clones were produced and used to study expression in developing barley and oat seeds. Actin and CDC48 mRNAs declined rapidly following inoculation of barley spikes with Fusarium graminearum. Despite this, permatin mRNA levels remained constant or increased slightly. Studies of permatin gene expression in healthy plants revealed that developing barley and oat seeds accumulate permatin mRNA in an unusual bimodal pattern. Permatin mRNA and protein are highly abundant around the time of pollination and then decrease rapidly to near-zero. A second peak occurs in the doughy stage of development. Antibody and DNA probe hybridization studies showed that expression initially occurs in the ovary wall and then switches to the aleurone and ventral furrow of developing seeds, reaching a peak in the doughy stage. Small amounts of permatin mRNAs also occur in certain vegetative tissues. The barley and oat permatin sequences provided sufficient comparisons between cereal TLPs to suggest that deletions or additions in specific elements could have led to the divergence of leaf- and seed-specific TLPs.
Journal Article↗
Gibberellin mobilizes distinct DELLA-dependent transcriptomes to regulate seed germination and floral development in Arabidopsis.
Severe Arabidopsis (Arabidopsis thaliana) gibberellin (GA)-deficient mutant ga1-3 fails to germinate and is impaired in floral organ development. In contrast, the ga1-3 gai-t6 rga-t2 rgl1-1 rgl2-1 mutant confers GA-independent seed germination and floral development. This fact suggests that GA-regulated transcriptomes for seed germination and floral development are DELLA dependent. However, it is currently not known if all GA-regulated genes are GA regulated in a DELLA-dependent fashion and if a similar set of DELLA-regulated genes is mobilized to repress both seed germination and floral development. Here, we compared the global gene expression patterns in the imbibed seeds and unopened flower buds of the ga1-3 mutant with that of the wild type and of the ga1-3 gai-t6 rga-t2 rgl1-1 rgl2-1 mutant. We found that about one-half of total GA-regulated genes are apparently regulated in a DELLA-dependent fashion, suggesting that there might be a DELLA-independent or -partially-dependent component of GA-dependent gene regulation. A cross-comparison based on gene identity revealed that the GA-regulated DELLA-dependent transcriptomes in the imbibed seeds and flower buds are distinct from each other. Detailed ontology analysis showed that, on one hand, DELLAs differentially regulate the expression of different individual members of a gene family to run similar biochemical pathways in seeds and flower. Meanwhile, DELLAs control many functionally different genes to run specific pathways in seeds or flower buds to mark the two different developmental processes. Our data shown here not only confirm many previous reports but also single out some novel aspects of DELLA functions that are instructive to our future research.
Amino Acid Sequence↗
Mitochondrial metabolism in developing embryos of Brassica napus.
The metabolism of developing plant seeds is directed toward transforming primary assimilatory products (sugars and amino acids) into seed storage compounds. To understand the role of mitochondria in this metabolism, metabolic fluxes were determined in developing embryos of Brassica napus. After labeling with [1,2-(13)C2]glucose + [U-(13)C6]glucose, [U-(13)C3]alanine, [U-(13)C5]glutamine, [(15)N]alanine, (amino)-[(15)N]glutamine, or (amide)-[(15)N]glutamine, the resulting labeling patterns in protein amino acids and in fatty acids were analyzed by gas chromatography-mass spectrometry. Fluxes through mitochondrial metabolism were quantified using a steady state flux model. Labeling information from experiments using different labeled substrates was essential for model validation and reliable flux estimation. The resulting flux map shows that mitochondrial metabolism in these developing seeds is very different from that in either heterotrophic or autotrophic plant tissues or in most other organisms: (i) flux around the tricarboxylic acid cycle is absent and the small fluxes through oxidative reactions in the mitochondrion can generate (via oxidative phosphorylation) at most 22% of the ATP needed for biosynthesis; (ii) isocitrate dehydrogenase is reversible in vivo; (iii) about 40% of mitochondrial pyruvate is produced by malic enzyme rather than being imported from the cytosol; (iv) mitochondrial flux is largely devoted to providing precursors for cytosolic fatty acid elongation; and (v) the uptake of amino acids rather than anaplerosis via PEP carboxylase determines carbon flow into storage proteins.
Acetyl Coenzyme A↗
[Brachytherapy for prostate cancer: old concept, new techniques].
Whereas it has been proposed almost one century ago, brachytherapy for prostate cancer has only recently emerged, especially thanks to endorectal echography, allowing to visualize seed implantation, to the development of seeds for permanent delivery and of micro-sources for high-dose rate delivery, and also to the development of three-dimension dosimetry programs allowing real-time implantations. For selected patients with localized prostate cancer (PSA < 10, Gleason < 7, no extracapsular extension, volume < 50-60 g), prostate brachytherapy with permanent implants (iodine 125 or, less frequently, palladium 103) gives results which appear at 10-15 years comparable to those of surgery. Incontinence and impotence rates appear lower than those of classical surgery. However, the first post-implant months are usually accompanied by urinary toxicity that should not be minimized. High-dose rate brachytherapy (HDR) could find its indications, in combination with conformal radiotherapy, in the treatment of more advanced forms, presenting an intermediate risk. It could also be an alternative to brachytherapy with permanent implants for the low-risk forms mentioned above, especially in developing countries where the cost of radioactive seeds slows down the use of this technique. Brachytherapy for prostate cancer should, therefore, find more and more indications, because of the increased incidence of prostate cancer, due to population ageing, of the increased proportion of localized forms, due to better detection, of the patient's request for less toxicity, and of the expecting lowering of the costs, which are now equivalent to those of surgery and should further lower.
Brachytherapy↗
Influence of seeding density and dynamic deformational loading on the developing structure/function relationships of chondrocyte-seeded agarose hydrogels.
Chondrocytes cultured in agarose hydrogels develop a functional extracellular matrix. Application of dynamic strain at physiologic levels to these constructs over time can increase their mechanical properties. In this study, the effect of seeding density (20 and 60 x 10(6) cells/ml) on tissue elaboration was investigated. Higher seeding densities increased tissue properties in free-swelling culture, with constructs seeded at 20 and 60 x 10(6) cells/ml reaching maximum values over the 63 day culture period of aggregate modulus HA: 43 +/- 15 kPa, Young's modulus EY: 39 +/- 3 kPa, and glycosaminglycan content [GAG]: 0.96% +/- 0.13% wet weight; and HA: 58 +/- 12 kPa, EY: 60 +/- 5 kPa, and [GAG]: 1.49% +/- 0.26% wet weight, respectively. It was further observed that the application of daily dynamic deformational loading to constructs seeded at 20 x 10(6) cells/ml enhanced biochemical content (approximately 150%) and mechanical properties (approximately threefold) compared to free-swelling controls by day 28. However, at a concentration of 60 x 10(6) cells/ml, no difference in mechanical properties was found in loaded samples versus their free-swelling controls. Multiple regression analysis showed that the mechanical properties of the tissue constructs depend more strongly on collagen content than GAG content; a finding that is more pronounced with the application of daily dynamic deformational loading. Our findings provide evidence for initial cell seeding density and nutrient accessibility as important parameters in modulating tissue development of engineered constructs, and their ability to respond to a defined mechanical stimulus.
Animals↗
Iron Transport to Developing Ovules of Pisum sativum (I. Seed Import Characteristics and Phloem Iron-Loading Capacity of Source Regions).
To understand the processes that control Fe transport to developing seeds, we have characterized seed growth and Fe accretion and have developed a radiotracer technique for quantifying phloem Fe loading in vegetative source regions of Pisum sativum. In hydroponically grown plants of cv Sparkle, developing ovules exhibited a seed-growth period of 22 d, with Fe import occurring throughout the 22-d period. Average Fe content of mature seeds was 19 [mu]g. Source tissues of intact plants were abraded and pulse labeled for 4 h with 100 [mu]M 59Fe(III)-citrate. Fe was successfully phloem loaded and transported to seeds from leaflets, stipules, and pod walls. Total export of 59Fe from labeled source regions was used to calculate tissue-loading rates of 36, 40, and 51 pmol of Fe cm-2 h-1 for the leaflet, stipule, and pod wall surfaces, respectively. By comparison, surface area measurements, along with seed-growth results, allowed us to calculate average theoretical influx values of 42 or 68 pmol of Fe cm-2 h-1 for vegetative tissues at nodes with one or two pods, respectively. Additional studies with the regulatory pea mutant, E107 (a single-gene mutant of cv Sparkle that can overaccumulate Fe), enabled us to increase Fe delivery endogenously to the vegetative tissues. A 36-fold increase in Fe content of E107 leaves, relative to Sparkle, resulted in no increase in Fe content of E107 seeds. Based on these findings, we hypothesized that Fe is phloem loaded in a chelated form, and the expression/synthesis of the endogenous chelator is an important factor in the control of Fe transport to the seeds.
Journal Article↗
Time course of lectin and storage protein biosynthesis in developing pea (Pisum sativum) seeds.
Lectins and storage proteins from pea (Pisum sativum) and other Leguminosae seeds interact in vitro. In order to evaluate whether this relationship may be important also in vivo, the time course of biosynthesis of these proteins in developing pea seeds was followed. The proteins appear in the following temporal order: vicilin-lectin-convicilin-legumin. The time course of protein synthesis was also followed by the subcellular morphological changes occurring simultaneously. The appearance of vicilin coincides with the formation of regularly shaped protein globules within the vacuole. Immediately after the start of lectin synthesis, these globules flatten and begin to adhere to the inner membrane surface of the vacuole. Legumin appears at a time when the vacuoles already have largely disappeared in favour of smaller vesicles which eventually develop into protein bodies. These results together with previous findings are consistent with the view that one of the biological functions of the lectin may consist in serving as a link between vicilin and the inner face of the vacuole membrane.
Antibodies, Monoclonal↗
The high oleate trait in the cultivated peanut [Arachis hypogaea L.]. I. Isolation and characterization of two genes encoding microsomal oleoyl-PC desaturases.
Plant oils rich in oleate are considered superior products compared to oils rich in polyunsaturated fatty acids. Peanut (Arachis hypogaea L.) is one of the major oilseed crops, and high oleate mutant varieties with as much as 85% oleate have been reported. We examined the possibility that this mutant phenotype resulted from reduction in the activity or the transcript level of microsomal oleoyl-PC desaturase. Two independently generated high oleate mutants, M2-225 and 8-2122, and their partially isogenic lines with a normal oleate phenotype were used in this study. Two cDNA sequences coding for microsomal oleoyl-PC desaturases, ahFAD2A and ahFAD2B, have been isolated from the developing peanut seed with a normal oleate phenotype. Cultivated peanut is an allotetraploid, and sequence comparisons with the genes from the putative diploid progenitor species suggested that ahFAD2A and ahFAD2B are non-allelic, but homeologous genes originating from two different diploid species. Northern analysis showed that the transcripts of oleoyl-PC desaturases are highly abundant in both normal and high oleate peanut seeds in the second stage of development. Differential digestion of the RT-PCR products revealed a restriction site polymorphism between ahFAD2A and ahFAD2B, and allowed us to examine the level of transcript expressed from each gene. The results indicate that ahFAD2A is expressed in both normal and high oleate peanut seeds, but the steady state level of the ahFAD2B transcript is severely reduced in the high oleate peanut varieties. These data suggested that the reduction in ahFAD2B transcript level in the developing seeds is correlated with the high oleate trait.
Amino Acid Sequence↗
A MADS box transcription factor of the AP1/AGL9 subfamily is also expressed in the seed coat of pea (Pisum sativum) during development.
As part of a programme to identify seed coat-specific transcripts in pea (Pisum sativum cv. Finale) using the differential display method, we identified and isolated a cDNA encoding a MADS box transcription factor. The predicted peptide contains 247 amino acids with the conserved N-terminal MADS box and is closely related to the AP1/AGL9 subfamily of the MADS box proteins. In addition to weak expression in petals, stamens and carpels, this MADS box mRNA is specifically expressed in the seed coat during seed development.
Amino Acid Sequence↗
Chemopreventive effects of pomegranate seed oil on skin tumor development in CD1 mice.
Pomegranate seed oil was investigated for possible skin cancer chemopreventive efficacy in mice. In the main experiment, two groups consisting each of 30, 4-5-week-old, female CD(1) mice were used. Both groups had skin cancer initiated with an initial topical exposure of 7,12-dimethylbenzanthracene and with biweekly promotion using 12-O-tetradecanoylphorbol 13-acetate (TPA). The experimental group was pretreated with 5% pomegranate seed oil prior to each TPA application. Tumor incidence, the number of mice containing at least one tumor, was 100% and 93%, and multiplicity, the average number of tumors per mouse, was 20.8 and 16.3 per mouse after 20 weeks of promotion in the control and pomegranate seed oil-treated groups, respectively (P <.05). In a second experiment, two groups each consisting of three CD(1) mice were used to assess the effect of pomegranate seed oil on TPA-stimulated ornithine decarboxylase (ODC) activity, an important event in skin cancer promotion. Each group received a single topical application of TPA, with the experimental group receiving a topical treatment 1 h prior with 5% pomegranate seed oil. The mice were killed 5 h later, and ODC activity was assessed by radiometric method. The experimental group showed a 17% reduction in ODC activity. Pomegranate seed oil (5%) significantly decreased (P <.05) tumor incidence, multiplicity, and TPA-induced ODC activity. Overall, the results highlight the potential of pomegranate seed oil as a safe and effective chemopreventive agent against skin cancer.
9,10-Dimethyl-1,2-benzanthracene↗
Digestion of legume starch granules by larvae of Zabrotes subfasciatus (Coleoptera: bruchidae) and the induction of alpha-amylases in response to different diets.
Zabrotes subfasciatus larvae possess three alpha-amylase isoforms as determined by in gel assays following SDS-PAGE. The two minor isoforms present lower electrophoretic mobility than the major form, and seem to occur as a heterodimer. When developed inside Vigna unguiculata (cowpea) seeds, fourth instar larvae have minor quantities of the slow-migrating forms, but when reared on seeds of Phaseolus vulgaris (common bean) or Phaseolus lunatus, the two slow-migrating forms are expressed in higher amounts, while activity of the major form was independent of the host seed. Larvae developing inside cowpea seeds at the beginning of the fourth instar were fed on flour from cotyledons of cowpea or common bean. Larvae fed on the common bean flour started to express the dimer in higher amounts when compared with the control larvae fed on cowpea flour. In an attempt to correlate differences between starch granules and the induction of alpha-amylases, a detailed study on the digestive process of the granules was conducted. Incorporation of purified starch granules into artificial diets did not induce the two minor alpha-amylases. The in vitro hydrolysis rates of purified granules and the pattern of dextrins liberated by the different alpha-amylases were similar for the two legume species. The starch granules enter the midgut extensively damaged, which may facilitate the access to the more susceptible parts of the granules to enzymatic attack.
Animals↗
Processing and localization of bovine beta-casein expressed in transgenic soybean seeds under control of a soybean lectin expression cassette.
We have examined the processing and subcellular localization of a chimeric gene consisting of the bovine milk protein, beta-casein, under the control of a soybean seed lectin promoter and its 32 amino acid signal sequence in the seeds of transgenic soybean plants. The beta-casein expressed in developing soybean seeds is a doublet with apparent molecular weight slightly smaller than the bovine beta-casein and expression of the protein was highest in immature cotyledons. The casein proteins were purified from the immature soybean seeds by immunoaffinity chromatography and were analyzed by two-dimensional gel electrophoresis, blotting, and amino terminal sequencing. The N-terminal sequences of both of the doublet soybean casein polypeptides were identical to the N-terminal sequence of the bovine beta-casein indicating that the 32 amino acid lectin signal sequence was cleaved precisely from the chimeric protein in developing soybean seeds. Analysis of the purified soybean beta-casein polypeptides by mass spectrometry (MALDI-MS) showed that they are not phosphorylated. Absence of added phosphate groups is the cause of the size difference between the soybean beta-casein and native bovine beta-casein protein. Immunolocalization experiments showed that the casein protein was found in the protein storage vacuoles (PSV) in developing and mature soybean seeds. The precise removal of the 32 amino acid lectin amino terminal sequence from the chimeric lectin-casein fusion suggests that the lectin expression cassette can be used for production of pharmaceutical or other recombinant proteins of added value in the developing soybean seed.
Journal Article↗
Cytosolic pyruvate kinase: subunit composition, activity, and amount in developing castor and soybean seeds, and biochemical characterization of the purified castor seed enzyme.
Antibodies against Brassica napus cytosolic pyruvate kinase (PKc) (EC 2.7.1.40) were employed to examine PKc subunit composition and developmental profiles in castor and soybean seeds. A 56-kDa immunoreactive polypeptide was uniformly detected on immunoblots of clarified extracts from developing castor endosperm or soybean embryos. Maximal PKc activities occurred early in castor oil seed (COS) and soybean development (7.1 and 5.5 (micromol of pyruvate produced/min) g(-1) FW, respectively) and were up to 25-fold greater than those of fully mature seeds. Time-course studies revealed a close correlation between extractable PKc activity and the relative amount of the immunoreactive 56-kDa PKc polypeptide. PKc from developing COS was purified 1,874-fold to homogeneity and a final specific activity of 73.1 (micromol of pyruvate produced/min) mg(-1) protein. Gel filtration and SDS-PAGE indicated that this PKc exists as a 230-kDa homotetramer composed of 56-kDa subunits. The mass fingerprint of tryptic peptides of the 56-kDa COS PKc subunit best matched three putative PK(c)s from Arabidopsis thaliana. The purified enzyme was relatively heat-stable and displayed a broad pH optimum of 6.4. However, more efficient substrate utilization (in terms of Vmax /Km for phosphoenolpyruvate or ADP) was observed at pH 7.4. Glutamate was the most effective inhibitor, whereas aspartate functioned as an activator by partially relieving glutamate inhibition. Together with our previous studies, the results: (1) allow a model to be formulated regarding the coordinate allosteric control of PKc and phosphoenolpyruvate carboxylase by aspartate and glutamate in developing COS, and (2) provide further biochemical evidence that castor plant PKc exists as tissue-specific isozymes that exhibit substantial differences in their respective physical and regulatory properties.
Allosteric Regulation↗
Effects of grape seed proanthocyanidins extract on mandibles in developing rats.
OBJECTIVE: Grape seed proanthocyanidins extract (GSPE), a flavonoid, has a beneficial effect on physical health, which may include the health of bone. The purpose of the present study was to investigate the effects of GSPE on mandibular bone by examining trabecular and cortical bone density, mineral content, and non-invasive bone strength in low-calcium diet rats. MATERIALS AND METHODS: Wistar male rats at 5 weeks old (n = 40) were divided into control (A), low-calcium diet (B), low-calcium diet plus standard diet (C), and low-calcium diet plus standard diet with supplementary GSPE (D) groups. Following 3 weeks of a calcium-restricted diet, group D rats were given 3 mg of GSPE as supplement in 100 g of a standard diet for the next 3 weeks. Following the 6-week experimental period, mandibular bones were examined using peripheral quantitative computed tomography (pQCT). RESULTS: There were no significant differences in body weight or trabecular bone area among the four groups. Trabecular bone density, and trabecular bone mineral content, cortical bone density, cortical bone cross-sectional area, and cortical bone mineral content were significantly higher in group D than in C. Further, Stress-strain index (SSI) values of xSSI and ySSI in group D were significantly higher than in C, although there was no significant difference in pSSI value between those two groups. CONCLUSION: Our results suggest that GSPE treatment caused an increase in both bone formation and bone strength in rat mandibles.
Animals↗
Different sets of cis-elements contribute to the expression of a catalase gene from castor bean during seed formation and postembryonic development in transgenic tobacco.
Deletion analysis of the promoter region of a gene for catalase, cat2, from castor bean (Ricinus communis) was performed to identify the cis-regulatory elements responsible for the expression of a beta-glucuronidase (GUS) fusion gene during seed formation and postembryonic development in transgenic tobacco. The analysis showed that multiple cis-elements contribute to the activity of the cat2 promoter during seed formation and postembryonic development. The 5'-upstream regions from -1,241 to -816 bp, from -720 to -682 bp, and from -632 to -535 bp, relative to the site of initiation of translation of cat2, contributed positively to the activity of the cat2 promoter during both stages. By contrast, the region from -816 to -720 bp had a negative effect at both stages. The region from -682 to -632 bp contributed positively to the activity during seed formation but negatively during postembyonic development. Histochemical analysis revealed that the multiple cis-elements determined not only the level of expression of the chimeric gene but also the tissue-specificity of such expression. For example, the region from -1,241 to -816 bp allowed expression of the chimeric gene in the axis of the embryo of the dry seed, as well as in the cortex of the middle part of the hypocotyl and at the base of epicotyl in the young seedling.
Base Sequence↗