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Sporulation in Bacillus subtilis 168. Comparison of alkaline phosphatase from sporulating and vegetative cells.

1. The purification of the ;vegetative' alkaline phosphatase of Bacillus subtilis 168 was simplified by ionic elution of the enzyme from intact cells. 2. The enzyme has a molecular weight of about 70000 and treatment of the enzyme with 10mm-hydrochloric acid or 6.0m-guanidine hydrochloride, beta-mercaptoethanol (0.1m) gives rise to enzymically inactive subunits. 3. The amino acid composition of the enzyme was determined. The N-terminal residue determined by the DNS chloride method is glycine. 4. The properties of this enzyme were compared with the ;sporulation' alkaline phosphatase of the same strain. 5. Although the ;sporulation' enzyme differs from the ;vegetative' enzyme in its physiology of appearance and apparent mRNA stability, an examination of properties of the enzymes revealed no differences. 6. The enzyme from both cell forms is bound to the particulate fraction of cell extracts, but can be solubilized by high concentrations of magnesium chloride; removal of the magnesium chloride, by dialysis, results in precipitation of both enzymes. Both enzymes can be removed from intact cells by ionic elution. 7. The ;vegetative' and ;sporulation' enzymes have identical pH optima, K(m) and K(i) values and electrophoretic mobilities in cellulose acetate. 8. Their half-life is 28min at 65 degrees C and their Q(10) is 1.25. 9. The molecular size determined by gel filtration on Sephadex G-100 is about 69000. 10. ;Vegetative' and ;sporulation' forms gave precipitin lines that were continuous and non-spurred when tested against antiserum prepared against the ;vegetative' enzyme. 11. The ;sporulation' alkaline phosphatase appears to be associated with stage II of sporulation and appears to be induced by something specifically concerned in sporulation and not by phosphate starvation.

Alkaline Phosphatase↗

Interactions between the ABI1 and the ectopically expressed ABI3 genes in controlling abscisic acid responses in Arabidopsis vegetative tissues.

In Arabidopsis, the abscisic acid (ABA)-Insensitive ABI1 and ABI3 genes have been proposed to act in separate ABA signalling cascades. Recessive mutations in ABI3 alter various physiological processes during seed development, whereas the dominant abi1 mutation inhibits ABA responses largely in vegetative tissues. The seed-specific ABI3 gene was ectopically expressed in the vegetative tissues of transgenic Arabidopsis plants carrying a transcriptional fusion between the CaMV 35S promoter and the ABI3 cDNA. Genetic interactions between the ectopically expressed ABI3 and endogenous ABI1 genes were investigated by monitoring diverse ABA responses in vegetative tissues. Ectopic expression of ABI3 conferred to plantlets the ability to accumulate the seed-specific At2S33 and AtEm1 mRNAs in response to ABA, and the abi1 mutation inhibited this ABI3-dependent induction of AtEm1 by ABA. Furthermore, ectopic expression of ABI3 also influenced ABI1-dependent responses that occur in wild-type vegetative tissues. Expression of ABI3 increased ABA induction of the Rab18 mRNA and ABA inhibition of root growth, and both responses were sensitive to the abi1 mutation in the presence as in the absence of ABI3. Finally, although ABI3 is thought to be a transcription activator and stomatal regulation is not known to involve transcriptional events, the ectopically expressed ABI3 gene suppressed the effect of the abi1 mutation on stomatal regulation. The present data demonstrate that ABI1 and ABI3 genetically interact in controlling diverse ABA responses in transgenic vegetative tissues. The possibility that the endogenous ABI1 and ABI3 genes may similarly act in a common ABA signalling pathway in seed is discussed with previous phenotypic studies of the abi1 and abi3 mutants.

Abscisic Acid↗

The liguleless2 gene of maize functions during the transition from the vegetative to the reproductive shoot apex.

During a maize plant's (Zea mays) development, the shoot apical meristem (SAM) generates an apex that proceeds through different phases: juvenile vegetative, adult vegetative and reproductive. During each phase the structures produced are distinguishable from structures produced during the other phases. In this paper, we demonstrate that the LIGULELESS2 (LG2) function is required for an accurate vegetative to reproductive phase transition. The maize gene liguleless2 (lg2) has been shown to encode a basic-leucine zipper (bZIP) protein and to function in narrowing the region from which the ligule and auricle develop in a typical maize leaf. Here we show that lg2 mutant plants can have reduced long tassel branches, extra vegetative leaves and extra husk leaves when compared to wild-type siblings. This indicates a role for the lg2 gene in the vegetative to reproductive phase transition of the shoot apex. We also discuss a potential role for the lg2 gene in general phase transition processes.

Alleles↗

Evidence for and localization of vegetative viral DNA replication by autoradiographic detection of RNA-DNA hybrids in sections of tumors induced by Shope papilloma virus.

The occurrence and localization of vegetative viral DNA replication was studied in sections of tumors induced by the rabbit Shope papilloma virus, in cottontail and domestic rabbit papillomas, in primary domestic rabbit carcinoma, and in transplantable VX2 carcinoma, by in situ hybridization of radioactive RNA complementary to viral DNA. Vegetative viral DNA replication and viral protein synthesis were compared by means of cytological hybridization and immunofluorescence techniques on adjacent frozen sections. Vegetative viral DNA replication is completely repressed in the proliferating cellular layers of these tumors, which suggests a provirus state of the viral genome, as in other cells transformed by oncogenic DNA viruses. Vegetative viral DNA replication is induced, after initiation of the keratinization, in cells of cottonail rabbit papillomas, where it is usually followed by viral protein synthesis; this illustrates the influence of the physiological state of the host cell on the control of viral functions. Vegetative viral DNA replication is deteced only in a few cells of domestic rabbit papillomas, at the end of the keratinization process; this observation provides indirect evidence that the DNA synthesis specifically induced in these tumors after the onset of keratinization reflects mostly the induction of cellular DNA synthesis.

Animals↗

A 69-kDa RNA-binding protein from Xenopus oocytes recognizes a common motif in two vegetally localized maternal mRNAs.

Vg1 mRNA, a maternal message encoding a member of the transforming growth factor beta superfamily, undergoes localization to the vegetal cortex of Xenopus laevis oocytes during a narrow period of oogenesis. A 340-nucleotide sequence has been identified in Vg1 RNA that directs its vegetal localization [Mowry, K. L. & Melton, D. A. (1992) Science 255, 991-994]. To understand how cis- and trans-acting factors are involved in Vg1 mRNA localization, we have looked for specific interactions in vitro between oocyte proteins and Vg1 mRNA. S100 extracts of late-stage oocytes contain a protein-binding activity that protects specific regions of labeled Vg1 mRNA from degradation by RNase T1. The use of different regions of Vg1 RNA in competition reactions reveals two binding sites, both in the first half of the 3' untranslated region of Vg1 message. UV crosslinking predominantly labels a 69-kDa protein; saturation analysis and competitor studies indicate that this protein binds with a high affinity to the down-stream site, which corresponds to the 340-nucleotide vegetal localization sequence. Binding to this region is inhibited by another vegetally localized message, transforming growth factor beta 5 but is not inhibited by an animally localized RNA, An2. These data indicate that vegetally localized mRNAs share a binding motif that helps them achieve their intracellular distribution through specific RNA-protein interactions.

Animals↗

Analysis of population structure of the chestnut blight fungus based on vegetative incompatibility genotypes.

Vegetative incompatibility is a self/nonself-recognition system in fungi that has often been used for describing phenotypic diversity in fungal populations. A common hypothesis is that vegetative incompatibility polymorphisms are maintained by balancing selection. However, understanding the evolutionary significance of vegetative incompatibility and the factors that maintain these polymorphisms has been limited by a lack of knowledge of the underlying genetics of vegetative compatibility (vc) types. Genotypes of 64 vc types, controlled by six unlinked vegetative incompatibility (vic) loci, have been identified in the chestnut blight fungus, Cryphonectria parasitica. By interpreting vc type survey data in terms of vic genotypes, we estimated vic-allele frequencies and analyzed the multilocus genetic structure of 13 populations in Europe and 3 populations in the U.S. European populations have less vc type diversity than the US populations because of a combination of lower vic-allele diversity and limited recombination. Genotypic diversity of 10 populations in Italy correlated to the abundance of sexual structures; however, significant deviations from random mating suggest that either sexual reproduction may not contribute many offspring in these populations or that vic genes (or vic genotypes) are under selection. Most vic-allele frequencies deviated from 0.5, the equilibrium frequency predicted under frequency-dependent selection, providing no evidence for selection acting on these loci.

Journal Article↗

Cruciferous vegetables and colo-rectal cancer.

Cruciferous vegetables have been studied extensively for their chemoprotective effects. Although they contain many bioactive compounds, the anti-carcinogenic actions of cruciferous vegetables are commonly attributed to their content of glucosinolates. Glucosinolates are relatively biologically inert but can be hydrolysed to a range of bioactive compounds such as isothiocyanates (ITC) and indoles by the plant-based enzyme myrosinase, or less efficiently by the colonic microflora. A number of mechanisms whereby ITC and indoles may protect against colo-rectal cancer have been identified. In experimental animals cruciferous vegetables have been shown to inhibit chemically-induced colon cancer. However, the results of recent epidemiological cohort studies have been inconsistent and this disparity may reflect a lack of sensitivity of such studies. Possible explanations for the failure of epidemiological studies to detect an effect include: assessment of cruciferous vegetable intake by methods that are subject to large measurement errors; the interaction between diet and genotype has not been considered: the effect that post-harvest treatments may have on biological effects of cruciferous vegetables has not been taken into account.

Anticarcinogenic Agents↗

The assessment of vegetative and functional symptoms in fibromyalgia patients: the tropisetron experience.

Vegetative and functional symptoms are, besides pain and tenderness of tender points, considered as additional information for the diagnosis of fibromyalgia (FM). In clinical trials, vegetative and functional symptoms have been included for selection of patients (e.g. sleep disturbances) and as secondary outcome parameters. Despite the relevance of these symptoms, no validated method is currently available but symptom lists are ad hoc developed by investigators. In this manuscript, data from a published double blind, randomised study are reanalysed which compared oral therapy over 10 days with 5 mg, 10 mg, and 15 mg to placebo in FM patients. This study applied a list of 17 vegetative and functional symptoms, which had to be scored by the patients by use of a 4-point severity scale (0 = none to 3 = severe). Factor analysis of the baseline data from 195 patients suggested to separate 6 sub-scales: Cardiovascular, gastrointestinal, psychiatric (sleep disturbance), nervous, autonomic system, and general disorders. Sleep disturbances, general symptoms (morning stiffness, fatigue) and autonomic symptoms (cold extremities, hyperhidrosis) were most severe in intensity. Analysis of sensitivity for treatment effects made use of differences between placebo and 5 mg tropisetron in changes between baseline and final assessment of the tropisetron trial. While, on the item level, differences in favour of tropisetron could only be demonstrated for sleep disorders, on the sub-scale level, also favourable effects of tropisetron could be shown for cardiovascular and nervous system complaints and, as a tendency, for general symptoms. On the other side, the sub-scale score of gastrointestinal symptoms worsened under tropisetron whilst it improved under placebo which effect was due to side effects of the active treatment. It is concluded that symptom clusters like sub-scales of a list of vegetative and functional symptoms will be more suitable for diagnostic purposes and evaluation of treatment outcome of clinical trials. Further research is urgently required which addresses the development of a FM-specific scale to assess vegetative and functional symptoms.

Fibromyalgia↗

Accumulation of metals in vegetation from an alkaline artificial soil.

This research was conducted to determine whether elevated concentrations of metals in biosolids would result in increased accumulation of these metals in plants growing on an alkaline, artificial soil created by addition of municipal sewage biosolids to inorganic waste materials during reclamation of a filled lime settling basin. Accumulation of metals in vegetation growing on this alkaline, artificial soil was compared with accumulation in vegetation growing on a natural, reference soil that had not been amended by biosolids. Although the concentrations of Cd, Cr, Hg, Ni, Ag, and Zn were greater in the artificial soil than in the reference soil, only Cd was found at greater concentrations in vegetation growing on the artificial soil. These greater concentrations were only observed in wheat grass (Agropyron sp.), and not in orchard grass (Dactylis glomerata L.). Also, the accumulation ratio, which is the ratio of the metal concentration in plant tissue to that in the soil, for wheat grass was much lower than other published values. There were no differences in the concentrations of Ba and Pb in the two soils, but the concentrations of these two metals in orchard grass growing on the reference soil were greater than that in either grass on the artificial soil. The differences in plant uptake of metals appear to result from differences in soil pH between the artificial and reference soils. Many metals are generally more soluble and bioavailable at the lower pH of the reference soil (pH = 5.4-6.6) than at the higher pH of the artificial soil (pH = 7.8-8.2). As a result, transfer of metals to vegetation growing on the alkaline, artificial soil is less than that in vegetation growing on the reference soil despite the greater concentrations of metals in the artificial soil.

Conservation of Natural Resources↗

Cover crops and natural vegetation mulch effect achieved by mechanical management with lateral rotary mower in weed population dynamics in citrus.

The aim of this study was to obtain information on practical weed management in order to reduce the use of herbicides thereby contributing to the sustainable development of citrus crop. The experiment was carried out under field conditions at the experimental area of the Department of Vegetal Production at the College of Agriculture "Luiz de Queiroz," Piracicaba, SP, Brazil, during the season 2002-2003. Influence of mulches produced by four types of vegetations on the dynamic population of weeds in the line of citrus crop (Citrus sinensis (L.) Osbeck) was evaluated. The experimental design was of factorial randomized blocks (4 x 2), where the treatments were: (i) four types of vegetation: Dolichos lablab L., Cajanus cajan (L.) Millsp, Penisetum glaucum (L.) Leeke, and the natural infestation composed basically by Panicum maximum Jacq.; and (ii) two types of fertilization: directed under canopy and broadcast. Mechanical management of the different vegetations was accomplished using a lateral rotary mower, KAMAQ, I model NINJA MAC 260, projected to release the green cut material under crop canopy, forming a mulch layer. The studied parameters were: (i) counting of weeds per m2 in the crop line after 30, 60, 90, 180, and 210 days following the cutting of existing vegetation; (ii) percentage of covered area by weeds; and (iii) some chemical properties of the soil. It was observed that the natural infestation showed a better weed control when compared with the other treatments, and that the broadcast fertilization, regardless of coverage used, presented a lower number of weeds.

Agriculture↗

Formation of vegetations during infective endocarditis excludes binding of bacterial-specific host antibodies to Enterococcus faecalis.

Infectious endocarditis is a microbial infection of the endothelial lining of the heart that typically occurs on damaged or prosthetic heart valves. The characteristic lesion seen with infective endocarditis, termed "the vegetation," is composed of bacteria surrounded by a platelet/fibrin layer attached to the underlying endothelium. The vegetation has long been believed to exclude or hinder host defenses from clearing bacteria, although formal demonstration of mechanisms by which this occurs are lacking. This study investigated the ability of the vegetation to exclude host antibodies specific for the bacterial surface protein aggregation substance in vivo during experimental endocarditis caused by Enterococcus faecalis. The results demonstrate that, once the vegetation encloses bacteria, they are no longer accessible to high-titer bacterial-specific host antibodies, establishing a mechanism by which the vegetation functions to protect the bacteria from the humoral immune response.

Animals↗

A high ratio of dietary animal to vegetable protein increases the rate of bone loss and the risk of fracture in postmenopausal women. Study of Osteoporotic Fractures Research Group.

BACKGROUND: Different sources of dietary protein may have different effects on bone metabolism. Animal foods provide predominantly acid precursors, whereas protein in vegetable foods is accompanied by base precursors not found in animal foods. Imbalance between dietary acid and base precursors leads to a chronic net dietary acid load that may have adverse consequences on bone. OBJECTIVE: We wanted to test the hypothesis that a high dietary ratio of animal to vegetable foods, quantified by protein content, increases bone loss and the risk of fracture. DESIGN: This was a prospective cohort study with a mean (+/-SD) of 7.0+/-1.5 y of follow-up of 1035 community-dwelling white women aged >65 y. Protein intake was measured by using a food-frequency questionnaire and bone mineral density was measured by dual-energy X-ray absorptiometry. RESULTS: Bone mineral density was not significantly associated with the ratio of animal to vegetable protein intake. Women with a high ratio had a higher rate of bone loss at the femoral neck than did those with a low ratio (P = 0.02) and a greater risk of hip fracture (relative risk = 3.7, P = 0.04). These associations were unaffected by adjustment for age, weight, estrogen use, tobacco use, exercise, total calcium intake, and total protein intake. CONCLUSIONS: Elderly women with a high dietary ratio of animal to vegetable protein intake have more rapid femoral neck bone loss and a greater risk of hip fracture than do those with a low ratio. This suggests that an increase in vegetable protein intake and a decrease in animal protein intake may decrease bone loss and the risk of hip fracture. This possibility should be confirmed in other prospective studies and tested in a randomized trial.

Absorptiometry, Photon↗

Identification of vib-1, a locus involved in vegetative incompatibility mediated by het-c in Neurospora crassa.

A non-self-recognition system called vegetative incompatibility is ubiquitous in filamentous fungi and is genetically regulated by het loci. Different fungal individuals are unable to form viable heterokaryons if they differ in allelic specificity at a het locus. To identify components of vegetative incompatibility mediated by allelic differences at the het-c locus of Neurospora crassa, we isolated mutants that suppressed phenotypic aspects of het-c vegetative incompatibility. Three deletion mutants were identified; the deletions overlapped each other in an ORF named vib-1 (vegetative incompatibility blocked). Mutations in vib-1 fully relieved growth inhibition and repression of conidiation conferred by het-c vegetative incompatibility and significantly reduced hyphal compartmentation and death rates. The vib-1 mutants displayed a profuse conidiation pattern, suggesting that VIB-1 is a regulator of conidiation. VIB-1 shares a region of similarity to PHOG, a possible phosphate nonrepressible acid phosphatase in Aspergillus nidulans. Native gel analysis of wild-type strains and vib-1 mutants indicated that vib-1 is not the structural gene for nonrepressible acid phosphatase, but rather may regulate nonrepressible acid phosphatase activity.

Acid Phosphatase↗

Evaluation of antibiotic diffusion into cardiac vegetations by quantitative autoradiography.

The reason bacterial endocarditis is difficult to cure has been controversial for many years. One explanation could be that antibiotic diffusion inside the vegetations is heterogeneous. This hypothesis was investigated by means of an autoradiographic study of diffusion of labeled antibiotics into large infected cardiac vegetations of nutritionally variant Streptococcus endocarditis in rabbits. Ten days after infection, 653 microCi of [3H]penicillin, 410 microCi of [3H]tobramycin, or 174 microCi of [14C]teicoplanin were injected iv over 30 min. Thirty minutes after the end of infusion (T30), vegetation/blood radioactivity ratios were 2.48 +/- 1.27, 2.49 +/- 0.67, and 3.94 +/- 1.19 for penicillin, tobramycin, and teicoplanin, respectively. Autoradiography clearly showed that distribution of the three drugs was different: Tobramycin was homogeneously distributed; penicillin was more concentrated at the periphery but still reached the center of vegetations; teicoplanin was concentrated only at the periphery. The same distribution pattern was observed with teicoplanin at T120 (i.e., one t1/2 beta later) and also after simultaneous infusion of a therapeutic dose (15 mg/kg) of cold teicoplanin. The diffusion gradient exhibited by some antibiotics could explain the difficulty in sterilizing vegetations despite high local concentrations, and the deleterious effect of the size of the vegetations on the therapeutic response.

Animals↗

Vegetable proteins enhance the growth of milk-fed piglets, despite lower apparent ileal digestibility.

This experiment compared the replacement of whey protein with isolated soy protein (ISP), or 2 levels of a hydrolyzed vegetable protein mixture (Lo HVPM and Hi HVPM, containing a partially hydrolyzed blend of soy, wheat, and other proteins) in liquid milk-replacer diets fed to neonatal pigs from 2 to 19 d of age. Piglets fed the vegetable protein diets weighed 20% more (8179 +/- 211 g, P < 0.05) at the end of the study than piglets fed the whey diet (6805 +/- 244 g). Growth rates were 35% higher for piglets fed the Hi HVPM diet than for piglets fed the whey diet. Similarly, intakes of the vegetable protein diets exceeded that for the whey diet (P < 0.05). Although the apparent ileal digestibilities of most amino acids were greater for the whey diet, digestible amino acid intakes (especially Arg, Phe, Met, and His) were greater in pigs fed the Hi HVPM and ISP diets (P < 0.01). Furthermore, carcasses of piglets fed the whey diet contained a higher percentage of fat and ash, whereas piglets fed the vegetable protein-containing diets accreted protein 42% faster (P < 0.01). Villus height and area and leucine aminopeptidase activity in the small intestine were greater in piglets fed the Lo HVPM diet than in those fed the ISP diet. Collectively, these data support the conclusion that some processed vegetable proteins may be good alternatives to whey protein in liquid diets formulated for neonatal pigs and that an appropriate balance of amino acids is more important than the source of protein per se.

Amino Acids↗

Uncertainty of historical measurements of 131I in Hanford-area vegetation.

The Hanford Environmental Dose Reconstruction (HEDR) Project was conducted to estimate the radiation dose that individuals could have received as a result of emissions to the air and water from Hanford Site operations since 1944. The largest doses were to the human thyroid gland from 131I released into the atmosphere from Hanford facilities in the 1945-1947 time period. In support of the dose reconstruction effort, a database of historical environmental radioactivity measurements was constructed. This database includes measurements of total radioactivity for vegetation samples collected from 1945-1948 and counted using a Geiger-Mueller (GM) detector system. Because the factors used at that time to convert the GM counts to 131I activity did not take all parameters into account, and because some parameter values were inaccurate, more accurate conversion factors were developed as part of the HEDR Project. These factors can be used to estimate the actual historical activity levels. This paper summarizes the Monte Carlo uncertainty and sensitivity analysis methods used to assess the uncertainty of the newly reconstructed historical vegetation 131I activities and to identify the parameters that contributed the most uncertainty to these reconstructed activities. Based on the study of two vegetation (sagebrush) samples collected in the mid-1940's, it appears that the true 131I activity of the historical vegetation samples should be within a factor of three of the reconstructed activity. Also, the uncertainty in the parameter Icf (the fraction of the background-corrected GM measurement of a vegetation sample that resulted from 131I) was found to contribute the most uncertainty to the reconstructed 131I activities when the uncertainty in Icf was large.

Air Pollutants, Radioactive↗

Long-term trends of radiocesium activity concentrations in vegetation in Irish semi-natural ecosystems.

137Cs activity concentrations were determined in vegetation from four Irish blanket bog ecosystems during the summer months of 1989-1997. The dominant and abundant vegetation species were sampled at each site and the data were used to estimate long-term trends of 137Cs activity concentrations. A general decline in the 137Cs activity concentrations in vegetation was observed from all sites sampled. The fastest and most consistent long-term rates of 137Cs decline were observed in the bryophyte and lichen vegetation group with 137Cs effective half-life (T ef) values of 2.2 to 10.7 y. The ericoid group with the exception of one case also showed a significant long-term decline in 137Cs activity concentrations with T ef values of 3.5 to 12.4 y as did the rush species Juncus squarrosus (T ef range 9.3 to 12.8 y). The sedge and grass groups showed the slowest and the least consistent rates of 137Cs decline, with T ef values of 2.9 to 59.8 y. Preliminary evaluation of 137Cs activity concentrations in sheep from some of the studied sites suggests that the decline found in vegetation is reflected in sheep.

Animals↗

Isolation and restriction analysis of DNA from heterocysts and vegetative cells of cyanobacteria.

A procedure was developed for the isolation of heterocysts from cyanobacterial filaments without recourse to mechanical disruption of the vegetative cells. DNA was then extracted from purified heterocysts by heating with 2% (w/v) SDS at 70 degrees C for 10 min. Following purification, this DNA was used for treatment with a range of restriction endonucleases and the results compared with DNA isolated from vegetative cells. Both heterocyst and vegetative DNAs from Anabaena PCC 7120 and Anabaena CA (ATCC 33047) were cut by XbaI, HindIII, EcoRI, ClaI, HpaII and MspI. However, none of the DNAs were cut by XhoI, SalI or MboI, indicating that the DNA from both organisms is methylated, but that no gross changes in methylation occur during heterocyst formation. Treatment of the DNAs with the former enzymes, followed by separation of the fragments by agarose gel electrophoresis, resulted in most cases in patterns of bands, which allowed a limited comparison of heterocyst and vegetative DNAs. No major differences were seen between the heterocyst and vegetative DNAs of either organism, implying that there are unlikely to be extensive rearrangements or major loss of DNA during heterocyst differentiation.

Cyanobacteria↗