Isolation of protoplast from soybean, cowpea, and tobacco and their fusion

International Nuclear Information System (INIS)

Irwansyah.

1988-01-01

Protoplast were isolated from leaf and callus. Young leaf of 3-4 weeks old plant of soybean T219 and A24, A27, C4, E1, and H6 of cowpeas strains (strains named by Prof. S. Sakamoto, University of Kyoto) were suspended in digestive medium containing cellulase 'Onuzuka' R-10, macerozyme R-10, mannitol, CaCl, and 2 (N-morpholilno) echane sulfonic acid (MES). For soybean leaf, the medium was enriched with driselase and pectolyase Y-23. They were incibated in full darkness at 27 Celcius centigrade by constant shaking at 50 rpm orbitor shaker. Callus wich has been two times resubcultured was suspended in the digestive medium without driselase, CaCl2, and MES and incubated in lowlight intensity by constant shaking at 100 rpm in reciprocal water shaker at 30 celcius centigrade. Leaf protoplast were releasaed in 10-14 h, soybean and tobacco callus protoplast in 3-4 h, and cowpeas callus protoplast in 4-6 h of incubation. Protoplast were collected by centrifugation of 400 g and a thin layer of the suspension was irradiated with ultraviolet light. Fusion was induced with PEG 6000 solution according to Uchimia and fused protoplasts were collected by centrifugation of 200 g. Protoplast were cultured on the medium of Ikeda and Uchimia. On both medium leaf protoplast, irradiated protoplasts and their fused do not regenerate cell wall and all cultured died out within four weeks incubation. Cell wall generation was observed. Regeneration of cell wall observed progessively in mother protoplast from tobacco, cowpea (A27, E1, and H6) and fused protoplast of soybean with tobacco, tobacco with cowpea (C4, E1, and H6), soybean with cowpea (C4) and between cowpea (C4) and cowpea (E1). (author). 25 refs, 4 tabs

Effect of microgravity environment on cell wall regeneration, cell divisions, growth, and differentiation of plants from protoplasts (7-IML-1)

Science.gov (United States)

Rasmussen, Ole

1992-01-01

The primary goal of this project is to investigate if microgravity has any influence on growth and differentiation of protoplasts. Formation of new cell walls on rapeseed protoplasts takes place within the first 24 hours after isolation. Cell division can be observed after 2-4 days and formation of cell aggregates after 5-7 days. Therefore, it is possible during the 7 day IML-1 Mission to investigate if cell wall formation, cell division, and cell differentiation are influenced by microgravity. Protoplasts of rapeseeds and carrot will be prepared shortly before launch and injected into 0.6 ml polyethylene bags. Eight bags are placed in an aluminum block inside the ESA Type 1 container. The containers are placed at 4 C in PTCU's and transferred to orbiter mid-deck. At 4 C all cell processes are slowed down, including cell wall formation. Latest access to the shuttle will be 12 hours before launch. In orbit the containers will be transferred from the PTC box to the 22 C Biorack incubator. The installation of a 1 g centrifuge in Biorack will make it possible to distinguish between effects of near weightlessness and effects caused by cosmic radiation and other space flight factors including vibrations. Parallel control experiments will be carried out on the ground. Other aspects of the experiment are discussed.

Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

Science.gov (United States)

Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

2016-01-01

The development of a range of sub-proteomic approaches to the plant cell wall has identified many of the cell wall proteins. However, it remains difficult to elucidate the precise biological role of each protein and the cell wall dynamics driven by their actions. The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. It therefore offers a unique opportunity to investigate the de novo construction process of the cell wall. This review deals with sub-proteomic approaches to the plant cell wall through the use of protoplasts, a methodology that will provide the basis for further exploration of cell wall proteins and cell wall dynamics. PMID:28248244

The retraction of the protoplast during PCD is an active, and interruptible, calcium-flux driven process.

Science.gov (United States)

Kacprzyk, Joanna; Brogan, Niall P; Daly, Cara T; Doyle, Siamsa M; Diamond, Mark; Molony, Elizabeth M; McCabe, Paul F

2017-07-01

The protoplast retracts during apoptosis-like programmed cell death (AL-PCD) and, if this retraction is an active component of AL-PCD, it should be used as a defining feature for this type of programmed cell death. We used an array of pharmacological and genetic tools to test if the rates of protoplast retraction in cells undergoing AL-PCD can be modulated. Disturbing calcium flux signalling, ATP synthesis and mitochondrial permeability transition all inhibited protoplast retraction and often also the execution of the death programme. Protoplast retraction can precede loss of plasma membrane integrity and cell death can be interrupted after the protoplast retraction had already occurred. Blocking calcium influx inhibited the protoplast retraction, reduced DNA fragmentation and delayed death induced by AL-PCD associated stresses. At higher levels of stress, where cell death occurs without protoplast retraction, blocking calcium flux had no effect on the death process. The results therefore strongly suggest that retraction of the protoplast is an active biological process dependent on an early Ca 2+ -mediated trigger rather than cellular disintegration due to plasma membrane damage. Therefore this morphologically distinct cell type is a quantifiable feature, and consequently, reporter of AL-PCD. Copyright © 2017 Elsevier B.V. All rights reserved.

Efficient production of Aschersonia placenta protoplasts for transformation using optimization algorithms.

Science.gov (United States)

Wei, Xiuyan; Song, Xinyue; Dong, Dong; Keyhani, Nemat O; Yao, Lindan; Zang, Xiangyun; Dong, Lili; Gu, Zijian; Fu, Delai; Liu, Xingzhong; Qiu, Junzhi; Guan, Xiong

2016-07-01

The insect pathogenic fungus Aschersonia placenta is a highly effective pathogen of whiteflies and scale insects. However, few genetic tools are currently available for studying this organism. Here we report on the conditions for the production of transformable A. placenta protoplasts using an optimized protocol based on the response surface method (RSM). Critical parameters for protoplast production were modelled by using a Box-Behnken design (BBD) involving 3 levels of 3 variables that was subsequently tested to verify its ability to predict protoplast production (R(2) = 0.9465). The optimized conditions resulted in the highest yield of protoplasts ((4.41 ± 0.02) × 10(7) cells/mL of culture, mean ± SE) when fungal cells were treated with 26.1 mg/mL of lywallzyme for 4 h of digestion, and subsequently allowed to recover for 64.6 h in 0.7 mol/L NaCl-Tris buffer. The latter was used as an osmotic stabilizer. The yield of protoplasts was approximately 10-fold higher than that of the nonoptimized conditions. Generated protoplasts were transformed with vector PbarGPE containing the bar gene as the selection marker. Transformation efficiency was 300 colonies/(μg DNA·10(7) protoplasts), and integration of the vector DNA was confirmed by PCR. The results show that rational design strategies (RSM and BBD methods) are useful to increase the production of fungal protoplasts for a variety of downstream applications.

Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro.

Science.gov (United States)

Thomas, Abitha; Pujari, Ipsita; Shetty, Vasudeep; Joshi, Manjunath B; Rai, Padmalatha S; Satyamoorthy, Kapaettu; Babu, Vidhu Sankar

2017-07-01

The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4% pectinase together with 2% cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 10 4 /ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.

Bicarbonate utilization by leaf protoplasts from Potamogeton

International Nuclear Information System (INIS)

Staal, M.; Elzenga, J.T.M.; Prins, H.B.A.

1987-01-01

Leaves from the submerged angiosperm P. lucens are able to assimilate bicarbonate. These leaves behave polarly: during bicarbonate utilization protons (H + ) are excreted by the cells of the lower epidermis, while hydroxyl (OH - ) ions are excreted by the upper epidermal cells. It has been proposed that acidification of the apoplast is a prerequisite for bicarbonate utilization. To test this hypothesis 14 C fixation by protoplasts was determined at different pH values. Also experiments, using the isotopic disequilibrium technique were performed. They showed that at pH values > 8, bicarbonate is a major carbon source for photosynthesis in protoplasts, despite the absence of cell walls and polarity. At pH values around 6, the rate of 14 C-fixation in protoplasts equals that of intact leaves. At pH values > 8, however, intact leaves show a higher rate. From this, and other experiments, the authors conclude that at least 2 processes contribute to bicarbonate utilization in P. lucens leaves: active transport (H + -HCO 3 - symport?) and acidification of the apoplast resulting in the conversion of bicarbonate into CO 2 . Polarity may increase the efficiency of both

Early Studies on Protoplast Isolation of Ludisia discolor, A Wild Orchid.

Science.gov (United States)

Poobathy, Ranjetta; Zakaria, Rahmad; Hamzah, Syed Mohd Edzham Syed; Subramaniam, Sreeramanan

2016-11-01

The terrestrial Ludisia discolor , also referred to as the jewel orchid is prized for the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heat- and pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures have emerged as alternative sources of anthocyanin production. Plant protoplast cultures are used frequently in transient gene expression studies and in the establishment of callus and cell suspension cultures. Benefits of plant protoplast system include similarity to cells found in plant tissues, reproduction under controlled conditions, and prevention of masking of stress responses to previous handling techniques. A study was conducted to assess the amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf segments were weighed, sliced into thin layers, immersed in a digestion medium, washed and then cultured onto a recovery medium. Results indicated that the production of plant protoplasts from L. discolor may be viewed as an alternative in the generation of cell cultures and ultimately in the production of anthocyanins from the cell cultures.

Isolation and culture of protoplast from leaves of Lactuca sativa

Directory of Open Access Journals (Sweden)

Witool Chaipakdee

2007-07-01

Full Text Available Protoplasts were isolated from leaves of lettuce (Lactuca sativa L. seedlings after in vitro germination for 25, 30, 40 and 50 days. The leaves were stripped and incubated in various combinations of cellulase and pectinase. Protoplasts were cultured on MS medium containing various kinds and concentrations of plant growth regulators in different culture systems including liquid media, hanging, drop culture and solid media. Results revealed that the highest number of viable protoplasts, 14.1x105 cells per gram of fresh weight, was obtained from 30 day-old leaves of lettuce seedlings and isolated by using 2% cellulase in combination with 1% pectinase. Liquid MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA promoted the highest cell division up to 17.67%. First division of protoplasts was observed at 4 days after culture and microcolony formation occurred at the 4th week after culturing. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

Plant regeneration from protoplasts of Gentiana straminea Maxim

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Shi Guomin

2016-01-01

Full Text Available A protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD and agar-pool (aPL culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D and 0.5 mg/L N6-benzylaminopurine (BA. Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

Proteins synthesized in tobacco mosaic virus infected protoplasts

International Nuclear Information System (INIS)

Huber, R.

1979-01-01

The author deals with research on the multiplication of tobacco mosaic virus (TMV) in leaf cell protoplasts. An attempt is made to answer three questions: (1) Which proteins are synthesized in TMV infected protoplasts as a result of TMV multiplication. (2) Which of the synthesized proteins are made under the direction of the TMV genome and, if any, which of the proteins are host specific. (3) In which functions are these proteins involved. (Auth.)

Protoplast formation and regeneration in Lactobacillus delbrueckii

OpenAIRE

Singhvi, Mamta; Joshi, Dipti; Gaikaiwari, Shalaka; Gokhale, Digambar V.

2010-01-01

Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and m...

Transformation of haploid, microspore-derived cell suspension protoplasts of rice (Oryza sativa L.).

Science.gov (United States)

Chaïr, H; Legavre, T; Guiderdoni, E

1996-06-01

We compared the transient activity of three cereal gene-derived promoter-gus fusions and the efficiency of selection mediated by three different selectable genes in a polyethylene glycol transformation system with haploid cell suspension protoplasts of rice. The maize ubiquitin promoter was found to be the most active in transformed protoplasts, and selection on ammonium glufosinate mediated by the bar gene was the most efficient for producing resistant calluses. Cotransformation of protoplasts with two separate plasmids carrying the gus and the bar genes, at either a 2∶1 or 1∶1 ratio, led to 0.8 × 10(-5) and 1.6 × 10(-5) resistant callus recovery frequencies and 59.7 and 37.9 cotransformation efficiencies respectively. No escapes were detected in dot blot analyses of 100 resistant calluses with a probe consisting of the bar coding region. Cotransformation efficiency, based on resistance to basta and β-glucuronidase staining of the leaf tissue of 115 regenerated plants, was 47%. Resistance tests and Southern analysis of seed progenies of three diploid transgenic plants demonstrated homozygous integration of multiple copies of the transgene at one locus at least in the first plant, heterozygous integration at one locus in the second plant and heterozygous integration at two loci in the third plant.

A study on the isolation of protoplasts from mesophyll cells of Dendrobium Queen Pink

International Nuclear Information System (INIS)

Aqeel, R.; Zehra, M.; Kazmi, S. K.; Khan, S.

2016-01-01

Protoplasts were successfully isolated from one month old In vitro grown plantlets of Dendrobium cultivar Queen pink. The enzyme solution used was composed of 1 percent Cellulase Onozuka R-10, 0.5 percent Macerozyme R-10, 0.1 percent Pectinase, 0.3 M mannitol, 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM 2 (N-morpholino)-ethanesulfonic acid (MES) at pH 5.8. Protoplast highest yield with 15.7x104 protoplasts per 1.5 gm freshly chopped leaves were obtained when digested in enzyme solution for 4 hrs on a rotary shaker with an agitation speed of 45 rpm in dark conditions. Protoplasts were filtered with 45 micro m nylon sieve and washed with 0.3 M mannitol solution supplemented with 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM MES, and purified with 0.3 M sucrose solution gradient. Purification of protoplasts on a sucrose mannitol gradient yielded clean protoplasts that were free from debris. (author)

Analysis of DNA polymerase activity in Petunia protoplasts treated with clastogenic agents

International Nuclear Information System (INIS)

Benediktsson, I.; Spampinato, C.P.; Andreo, C.S.; Schieder, O.

1994-01-01

Clastogenic agents, i.e. agents that can induce chromosome or DNA breakage, have been shown to enhance the role of direct gene transfer to protoplasts. The effect was analysed at the enzymatic level using protoplast homogenates as well as intact protoplasts. For that purpose existing procedures were modified to enable measurement of DNA polymerase in vivo. In the system used, external DNA was able to enter the cells without the addition of membrane-permeabilizing compounds. When comparing total DNA polymerase activity of protoplasts irradiated with X-rays or UV-light with that of untreated cells we did not observe significant differences. Incubation of protoplasts with high doses of bleomycin affected total DNA polymerase activity negatively. but dideoxythymidine triphosphate-sensitive activity was not influenced. We conclude that the DNA strand-breaks induced by low doses of X-rays. UV-light or bleomycin do not increase the total or the repair-DNA polymerase activity and. therefore. that the increase in the transformation rates after DNA strand-breaking is not preceded by enhanced DNA polymerase activity. (author)

Formation and regeneration of protoplasts and spheroplasts of gastrointestinal strains of lactobacilli.

OpenAIRE

Connell, H; Lemmon, J; Tannock, G W

1988-01-01

Methods were developed for the formation of protoplasts and spheroplasts of gastrointestinal strains of Lactobacillus reuteri, Lactobacillus gasseri, and Lactobacillus salivarius. Attempts to regenerate vegetative cells from protoplasts were not successful, but spheroplasts could be regenerated consistently for five of six strains.

Microcolony formation from embryogenic callus-derived protoplasts of oil palm

Directory of Open Access Journals (Sweden)

Sompong Te-chato

2005-07-01

Full Text Available Embryogenic callus of oil palm induced from young leaves of seedlings DxP was used as initial material for protoplast isolation. Various combinations of cellulase Onozuka RS and macerozyme R-10 were tested. Isolated protoplasts were cultured by various methods in MS medium supplemented with different phytohormones. The results revealed that 2% cellulase RS in combination with 2% macerozyme R-10 (adjusted osmoticum to 0.4 M by manitol yielded the highest number of viable protoplasts (1x107 per gram fresh weight. Dicamba at concentration 2 mg/l with 1 mg/l 6-benzyladenin (BA containing in phytagel semisolidified MS medium promoted the highest division of 2.3-4.0%. First division of the protoplasts was observed at 4 days after culture. Microcolony formation (8-10 cells was seen after three weeks of culture. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

Intergenus Protoplast Fusion between Pichia manshurica and Rhodosporidium paludigenum to Increase the Production of Inulinase

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Wijanarka Wijanarka

2014-12-01

Full Text Available The purposes of this study was to identify the optimum concentration of the lytic enzyme Glucanex for protoplast isolation and to conduct fusion for the purpose of increasing inulinase production. The study performs the protoplast fusion technique using Pichia manshurica and Rhodosporidium paludigenum. Protoplast fusion consists of a series of stages: protoplast isolation, protoplast fusion, protoplast regeneration, and analysis of hybrid fusion results. Protoplast isolation and fusion success rate are determined by various factors, including age of the culture, media type, and type of lytic enzymes used. Hybrid results were analyzed using a fungicide as a marker and measuring specific growth rate (μ of the hybrid compared with parental growth rates. Results demonstrated that a concentration of 4 mg/mL of Glucanex produces the greatest number of protoplasts, 7.2 x 1010 (cell/mL for P. manshurica and 8.8 x 1010 (cell/mL for Rh. paludigenum. The results of analysis of hybrid fusions indicate that the study has identified a new fusant, called fusant F4. Fusant F4 is capable of producing the highest inulinase, 0.6892 IU, compared with parentals P. manshurica, 0557 IU, and Rh. paludigenum, 0.3263 IU. Fusant F4 has specific growth rate (μ of 0.3360/h and generation time (g of 2.0629 h.

Isolation and culture of leaf protoplasts from Tunisian grapes

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Ahmed Mliki

2003-09-01

Full Text Available Experimental conditions for leaf protoplast isolation and culture were optimised for in vitro plants deriving from shoot culture of two Tunisian grape varieties, Sakasly and Muscat d’Alexandrie (Vitis vinifera L.. The best yields were obtained from leaves of 4 to 5 weeks old in vitro plants, digested for 13 hours under 25 rpm agitation with an enzymatic mixture containing 0.25 % cellulase of Aspergillus niger, 0.25 % cellulase of Penicillium funiculosum, 0.5 % cellulysin of Trichoderma viridae, and 0.2 % macerozyme R-10 of Rhizopus sp. More than 50 % of the purified protoplasts had a diameter of 30-40 μm and were rich in chloroplasts. Best aptitude for cell division was found in protoplasts immobilised in sodium alginate layers at a density of 0.5x106 cell/ml, cultivated in CPW-13 medium containing 4 mg/l of NOA and 0.88 mg/l of TDZ. The variety Muscat d’Alexandrie gave better yield whereas Sakasly showed better cell division rates. Formation of micro and macrocallus have been obtained, but the oxidation of the medium has to be solved in order to promote plant regeneration.

14-3-3 Proteins in Guard Cell Signaling.

Science.gov (United States)

Cotelle, Valérie; Leonhardt, Nathalie

2015-01-01

Guard cells are specialized cells located at the leaf surface delimiting pores which control gas exchanges between the plant and the atmosphere. To optimize the CO2 uptake necessary for photosynthesis while minimizing water loss, guard cells integrate environmental signals to adjust stomatal aperture. The size of the stomatal pore is regulated by movements of the guard cells driven by variations in their volume and turgor. As guard cells perceive and transduce a wide array of environmental cues, they provide an ideal system to elucidate early events of plant signaling. Reversible protein phosphorylation events are known to play a crucial role in the regulation of stomatal movements. However, in some cases, phosphorylation alone is not sufficient to achieve complete protein regulation, but is necessary to mediate the binding of interactors that modulate protein function. Among the phosphopeptide-binding proteins, the 14-3-3 proteins are the best characterized in plants. The 14-3-3s are found as multiple isoforms in eukaryotes and have been shown to be involved in the regulation of stomatal movements. In this review, we describe the current knowledge about 14-3-3 roles in the regulation of their binding partners in guard cells: receptors, ion pumps, channels, protein kinases, and some of their substrates. Regulation of these targets by 14-3-3 proteins is discussed and related to their function in guard cells during stomatal movements in response to abiotic or biotic stresses.

IP3 stimulates CA++ efflux from fusogenic carrot protoplasts

International Nuclear Information System (INIS)

Rincon, M.; Boss, W.F.

1986-01-01

Polyphosphoinositide breakdown plays an important role in signal transduction in animal cells (Berridge and Irvine, 1984, Nature, 312:315). Upon stimulation, phospholipase C hydrolyzes phosphatidylinositol 4,5-bisphosphate to inositol 1,4,5-trisphosphate (IP 3 ) and diacylglycerol both of which act as cellular second messengers. IP 3 mobilizes Ca ++ from internal stores, hence the cytosolic free Ca ++ concentration increases and those physiological activities regulated by Ca ++ are stimulated. To test if plant cells also responded to IP 3 , Ca ++ efflux studies were done with fusogenic carrot protoplasts released in EGTA. The protoplasts were preloaded with 45 Ca ++ placed in a Ca ++ -free medium, and efflux determined as 45 Ca ++ loss from the protoplasts. IP 3 (10-20μM) caused enhanced 45 Ca ++ efflux and the response was sustained for at least 15 min. In plants, as in animals, the observed IP 3 -enhanced 45 Ca ++ efflux suggested that IP 3 released Ca ++ from internal stores, and the increased free cytosolic Ca ++ activated Ca ++ pumping mechanisms which restored the Ca ++ concentration in the cytosol to the normal level

Effect of X-Rays on Growth Rate of Rose Shoot Cultures and the Ability of Isolated Protoplasts to Form Cell Colonies

International Nuclear Information System (INIS)

Moustafa, R. A. K.

2004-01-01

The popularity of rose as a garden plant, allied with its use in the production of cut flowers and also as a source of aromatic rose oils, make it one of the most important ornamental crops. Roses, however, have suffered from a narrow genetic base to which only few species have contributed significantly. In vitro culture of plants might facilitate the improvement of rose via the exploitation of somaclonal variation to generate new genetic variability and selection within the variation for desirable traits. The application of mutagens for in vitro cultures, in addition to the induced mutations, may lead to increase the somaclonal variation, thus providing additional variation for selection. On the other hand, plant protoplasts offer exciting possibilities to establish in vitro selection programs based on single cells. Induced variation in isolated protoplasts using mutagen agents may be one mean to select useful mutants. Thus the present experiments were conducted to determine the effect of X-rays on shoot cultures and the isolated protoplasts of rose (Rosa sp.). The materials consisted of the three rose varieties Rosa wichuriana, Paricer charm and Heckenzauber.The applied doses were 0, 10, 20, 30, 40, 50 and 60 Gy. Obtained results indicated that the genotypes differed in their sensitivity to X-rays. Rosa wichuriana seemed to be the most sensitive variety to radiation, where a dose of 20 Gy caused approximately 50% reduction in growth rate of shoot cultures, while the same dose decreased the growth rate of Paricer charm only by 25% and did not affect the growth of Heckenzauber. Results also revealed that the ability of irradiated protoplasts to form cell colonies increased when a dose of 10 Gy was performed. Doses higher than that level caused gradual decreasing in the forming of cell colonies, but however, the protoplasts could form colonies even when a dose of 60 Gy was applied. (Author)

Ability of Bacillus subtilis protoplasts to repair irradiated bacteriophage deoxyribonucleic acid via acquired and natural enzymatic systems

International Nuclear Information System (INIS)

Yasbin, R.E.; Andersen, B.J.; Sutherland, B.M.

1981-01-01

A novel form of enzyme therapy was achieved by utilizing protoplasts of Bacillus subtilis. Photoreactivating enzyme of Escherichia coli was successfully inserted into the protoplasts of B. subtilis treated with polyethylene glycol. This enzyme was used to photoreactivate ultraviolet-damaged bacteriophage deoxyribonucleic acid (DNA). Furthermore, in polyethylene glycol-treated protoplasts, ultraviolet-irradiated transfecting bacteriophage DNA was shown to be a functional substrate for the host DNA excision repair system. Previous results (R.E. Yasbin, J.D. Fernwalt, and P.I. Fields, J. Bacteriol.; 137: 391-396) showed that ultraviolet-irradiated bacteriophage DNA could not be repaired via the excision repair system of competent cells. Therefore, the processing of bacteriophage DNA by protoplasts and by competent cells must be different. This sensitive protoplast assay can be used to identify and to isolate various types of DNA repair enzymes

A simple and effective method to encapsulate tobacco mesophyll protoplasts to maintain cell viability

Directory of Open Access Journals (Sweden)

Rong Lei

2015-01-01

• It is very convenient to change or collect the solution without mechanically disturbing the protoplasts. This simple and effective silica sol–gel/alginate two-step immobilization of protoplasts in Transwell has great potential for applications in genetic transformation, metabolite production, and migration assays.

Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

OpenAIRE

Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

2016-01-01

The development of a range of sub-proteomic approaches to the plant cell wall has identified many of the cell wall proteins. However, it remains difficult to elucidate the precise biological role of each protein and the cell wall dynamics driven by their actions. The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. It therefore offers a uni...

Efficient Transformation of Oil Palm Protoplasts by PEG-Mediated Transfection and DNA Microinjection

Science.gov (United States)

Masani, Mat Yunus Abdul; Noll, Gundula A.; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

2014-01-01

Background Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants. Methodology/Principal Findings We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation. Conclusions/Significance We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants. PMID:24821306

Early activation of lipoxygenase in lentil (Lens culinaris) root protoplasts by oxidative stress induces programmed cell death

NARCIS (Netherlands)

Vliegenthart, J.F.G.; Maccarrone, M.; Zadelhoff, G. van; Veldink, G.A.; Finazzi Agrò, A.

2000-01-01

Oxidative stress caused by hydrogen peroxide (H2O2) triggers the hypersensitive response of plants to pathogens. Here, short pulses of H2O2 are shown to cause death of lentil (Lens culinaris) root protoplasts. Dead cells showed DNA fragmentation and ladder formation, typical hallmarks of apoptosis

Use of protoplast, cell, and shoot tip culture in an elm germ plasm improvement program

Science.gov (United States)

R. Daniel Lineberger; M.B. Sticklen; P.M. Pijut; Mark A. Kroggel; C.V.M. Fink; S.C. Domir

1990-01-01

An elm germplasm improvement program was established using three distinct approaches: (1) development of protoplast regeneration protocols with the goal of attempting somatic hybridization between Ulmus americana and disease resistant hybrids; (2) evaluation of the extent of somaclonal variation in plants regenerated from protoplasts; and (3)...

Radiosensitivity of protoplasts of orange (Citrus sinensis)

International Nuclear Information System (INIS)

Goldman, M.H.S.; Ando, A.

1990-01-01

Full text: The Radiation Genetics Section of the Centre for Nuclear Energy in Agriculture (CENA), University of Sao Paulo, is utilising both ''in vivo'' and ''in vitro'' methods for mutation induction in Citrus, cv. ''Pera'', aiming at resistance to citrus canker. The experiments carried out so far determined the methodology to isolate protoplasts and their sensitivity to gamma-rays. Regarding the culture of protoplasts from embryogenic callus, the best experimental conditions were: enzymatic digestion for 5 h on a medium containing cellulase (307.6 mg/10 ml), macerozyme (30.3 mg/10 ml), mannitol (328.0 mM) and sucrose (336.2 mM) as osmotic stabilisers. The isolation efficiency of 1.2x10 6 viable protoplasts/g will make it possible to use protoplasts in mutation breeding. To determine radiosensitivity of protoplasts, gamma-irradiation from 60 Co source was conducted 42 h after their isolation. This time interval is recommended because during this period protoplasts will reach the stage prior to or at the first mitotic division. Survivals were determined by metylen-blue dyeing, and the LD 50 was found to be around 37.5 Gy. Any difference compared with other authors might be due to different genotypes used or different methods of calculation of survival. (author)

Formation and cell wall regeneration of protoplasts from Schizophyllum commune

NARCIS (Netherlands)

de Vries, Onno Minne Hotze

1974-01-01

Osmotically sensitive protoplasts were released from the mycelium of the basidiomycete Schizophyllum commune through the action ofan extracellular enzyme preparation isolated from the culture filtrate of Trichoderma viride (recently renamed T. harzianum) grown on hyphal walls of the former organism.

Nitric Oxide (NO) Measurements in Stomatal Guard Cells.

Science.gov (United States)

Agurla, Srinivas; Gayatri, Gunja; Raghavendra, Agepati S

2016-01-01

The quantitative measurement of nitric oxide (NO) in plant cells acquired great importance, in view of the multifaceted function and involvement of NO as a signal in various plant processes. Monitoring of NO in guard cells is quite simple because of the large size of guard cells and ease of observing the detached epidermis under microscope. Stomatal guard cells therefore provide an excellent model system to study the components of signal transduction. The levels and functions of NO in relation to stomatal closure can be monitored, with the help of an inverted fluorescence or confocal microscope. We can measure the NO in guard cells by using flouroprobes like 4,5-diamino fluorescein diacetate (DAF-2DA). This fluorescent dye, DAF-2DA, is cell permeable and after entry into the cell, the diacetate group is removed by the cellular esterases. The resulting DAF-2 form is membrane impermeable and reacts with NO to generate the highly fluorescent triazole (DAF-2T), with excitation and emission wavelengths of 488 and 530 nm, respectively. If time-course measurements are needed, the epidermis can be adhered to a cover-glass or glass slide and left in a small petri dishes. Fluorescence can then be monitored at required time intervals; with a precaution that excitation is done minimally, only when a fluorescent image is acquired. The present method description is for the epidermis of Arabidopsis thaliana and Pisum sativum and should work with most of the other dicotyledonous plants.

Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool

Science.gov (United States)

Yang, Yingzhen; Costa, Alex; Leonhardt, Nathalie; Siegel, Robert S; Schroeder, Julian I

2008-01-01

Background A common limitation in guard cell signaling research is that it is difficult to obtain consistent high expression of transgenes of interest in Arabidopsis guard cells using known guard cell promoters or the constitutive 35S cauliflower mosaic virus promoter. An additional drawback of the 35S promoter is that ectopically expressing a gene throughout the organism could cause pleiotropic effects. To improve available methods for targeted gene expression in guard cells, we isolated strong guard cell promoter candidates based on new guard cell-specific microarray analyses of 23,000 genes that are made available together with this report. Results A promoter, pGC1(At1g22690), drove strong and relatively specific reporter gene expression in guard cells including GUS (beta-glucuronidase) and yellow cameleon YC3.60 (GFP-based calcium FRET reporter). Reporter gene expression was weaker in immature guard cells. The expression of YC3.60 was sufficiently strong to image intracellular Ca2+ dynamics in guard cells of intact plants and resolved spontaneous calcium transients in guard cells. The GC1 promoter also mediated strong reporter expression in clustered stomata in the stomatal development mutant too-many-mouths (tmm). Furthermore, the same promoter::reporter constructs also drove guard cell specific reporter expression in tobacco, illustrating the potential of this promoter as a method for high level expression in guard cells. A serial deletion of the promoter defined a guard cell expression promoter region. In addition, anti-sense repression using pGC1 was powerful for reducing specific GFP gene expression in guard cells while expression in leaf epidermal cells was not repressed, demonstrating strong cell-type preferential gene repression. Conclusion The pGC1 promoter described here drives strong reporter expression in guard cells of Arabidopsis and tobacco plants. It provides a potent research tool for targeted guard cell expression or gene silencing. It is also

Exploring emergent properties in cellular homeostasis using OnGuard to model K+ and other ion transport in guard cells.

Science.gov (United States)

Blatt, Michael R; Wang, Yizhou; Leonhardt, Nathalie; Hills, Adrian

2014-05-15

It is widely recognized that the nature and characteristics of transport across eukaryotic membranes are so complex as to defy intuitive understanding. In these circumstances, quantitative mathematical modeling is an essential tool, both to integrate detailed knowledge of individual transporters and to extract the properties emergent from their interactions. As the first, fully integrated and quantitative modeling environment for the study of ion transport dynamics in a plant cell, OnGuard offers a unique tool for exploring homeostatic properties emerging from the interactions of ion transport, both at the plasma membrane and tonoplast in the guard cell. OnGuard has already yielded detail sufficient to guide phenotypic and mutational studies, and it represents a key step toward 'reverse engineering' of stomatal guard cell physiology, based on rational design and testing in simulation, to improve water use efficiency and carbon assimilation. Its construction from the HoTSig libraries enables translation of the software to other cell types, including growing root hairs and pollen. The problems inherent to transport are nonetheless challenging, and are compounded for those unfamiliar with conceptual 'mindset' of the modeler. Here we set out guidelines for the use of OnGuard and outline a standardized approach that will enable users to advance quickly to its application both in the classroom and laboratory. We also highlight the uncanny and emergent property of OnGuard models to reproduce the 'communication' evident between the plasma membrane and tonoplast of the guard cell. Copyright © 2014 The Authors. Published by Elsevier GmbH.. All rights reserved.

Guard cell zeaxanthin tracks photosynthetically active radiation and stomatal apertures in Vicia faba leaves

International Nuclear Information System (INIS)

Srivastava, A.; Zeiger, E.

1995-01-01

Zeaxanthin, antheraxanthin and violaxanthin concentrations in guard cells from sonicated abaxial epidermal peels of Vicia faba were measured from dawn to dusk, and compared with concentrations in mesophyll tissue of the same leaves. Measured changes in guard cell zeaxanthin and violaxanthin concentrations indicate that guard cells operate the xanthophyll cycle throughout the day. Mesophyll tissue had no detectable zeaxanthin at dawn, whereas guard cells had 30–50 mmol mol −1 chlorophyll a+b. On a chlorophyll basis, maximal zeaxanthin levels were 3–4 fold higher in guard cells than in mesophyll cells. Zeaxanthin concentrations tracked levels of photosynthetically active radiation (PAR) in both mesophyll and guard cells. In the mesophyll, most of the zeaxanthin changes occurred in mid-morning and mid-afternoon. In guard cells, zeaxanthin concentrations changed nearly linearly with PAR in the early morning and late afternoon, and closely tracked PAR levels throughout the day. Guard cell zeaxanthin concentrations were also closely correlated with stomatal apertures. The close relationship between zeaxanthin concentrations and PAR levels in guard cells indicates that zeaxanthin is well suited to function as a molecular photosensor in stomatal movements. (author)

Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool

Directory of Open Access Journals (Sweden)

Siegel Robert S

2008-02-01

Full Text Available Abstract Background A common limitation in guard cell signaling research is that it is difficult to obtain consistent high expression of transgenes of interest in Arabidopsis guard cells using known guard cell promoters or the constitutive 35S cauliflower mosaic virus promoter. An additional drawback of the 35S promoter is that ectopically expressing a gene throughout the organism could cause pleiotropic effects. To improve available methods for targeted gene expression in guard cells, we isolated strong guard cell promoter candidates based on new guard cell-specific microarray analyses of 23,000 genes that are made available together with this report. Results A promoter, pGC1(At1g22690, drove strong and relatively specific reporter gene expression in guard cells including GUS (beta-glucuronidase and yellow cameleon YC3.60 (GFP-based calcium FRET reporter. Reporter gene expression was weaker in immature guard cells. The expression of YC3.60 was sufficiently strong to image intracellular Ca2+ dynamics in guard cells of intact plants and resolved spontaneous calcium transients in guard cells. The GC1 promoter also mediated strong reporter expression in clustered stomata in the stomatal development mutant too-many-mouths (tmm. Furthermore, the same promoter::reporter constructs also drove guard cell specific reporter expression in tobacco, illustrating the potential of this promoter as a method for high level expression in guard cells. A serial deletion of the promoter defined a guard cell expression promoter region. In addition, anti-sense repression using pGC1 was powerful for reducing specific GFP gene expression in guard cells while expression in leaf epidermal cells was not repressed, demonstrating strong cell-type preferential gene repression. Conclusion The pGC1 promoter described here drives strong reporter expression in guard cells of Arabidopsis and tobacco plants. It provides a potent research tool for targeted guard cell expression or

Stomatal Function Requires Pectin De-methyl-esterification of the Guard Cell Wall.

Science.gov (United States)

Amsbury, Sam; Hunt, Lee; Elhaddad, Nagat; Baillie, Alice; Lundgren, Marjorie; Verhertbruggen, Yves; Scheller, Henrik V; Knox, J Paul; Fleming, Andrew J; Gray, Julie E

2016-11-07

Stomatal opening and closure depends on changes in turgor pressure acting within guard cells to alter cell shape [1]. The extent of these shape changes is limited by the mechanical properties of the cells, which will be largely dependent on the structure of the cell walls. Although it has long been observed that guard cells are anisotropic due to differential thickening and the orientation of cellulose microfibrils [2], our understanding of the composition of the cell wall that allows them to undergo repeated swelling and deflation remains surprisingly poor. Here, we show that the walls of guard cells are rich in un-esterified pectins. We identify a pectin methylesterase gene, PME6, which is highly expressed in guard cells and required for stomatal function. pme6-1 mutant guard cells have walls enriched in methyl-esterified pectin and show a decreased dynamic range in response to triggers of stomatal opening/closure, including elevated osmoticum, suggesting that abrogation of stomatal function reflects a mechanical change in the guard cell wall. Altered stomatal function leads to increased conductance and evaporative cooling, as well as decreased plant growth. The growth defect of the pme6-1 mutant is rescued by maintaining the plants in elevated CO 2 , substantiating gas exchange analyses, indicating that the mutant stomata can bestow an improved assimilation rate. Restoration of PME6 rescues guard cell wall pectin methyl-esterification status, stomatal function, and plant growth. Our results establish a link between gene expression in guard cells and their cell wall properties, with a corresponding effect on stomatal function and plant physiology. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

Energy Technology Data Exchange (ETDEWEB)

Bona, Claudine Maria de [Instituto Agronomico do Parana (IAPAR), Curitiba, PR (Brazil)], e-mail: debona@iapar.br; Gould, Jean Howe [Texas A and M University, College Station, TX (United States). Dept. of Ecosystem Science and Management], e-mail: gould@tamu.edu; Miller Junior, J. Creighton [Texas A and M University, College Station, TX (United States). Dept. of Horticultural Sciences], e-mail: jcmillerjr@tamu.edu; Stelly, David [Texas A and M University, College Station, TX (United States). Dept. of Soil and Crop Sciences], e-mail: stelly@tamu.edu; Louzada, Eliezer Silva [Texas A and M University, Kingsville, TX (United States). Citrus Center], e-mail: e-louzada@tamu.edu

2009-05-15

The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma.irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaborai', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L{sup -1} iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol.butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNA samples. The best treatment was the donor-recipient fusion combination of 80 Gy.irradiated 'Ruby Red' protoplasts with 20 min IOA.treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L{sup -1} IBA solution for 10 min. (author)

Naturally induced secretions of the potato cyst nematode co-stimulate the proliferation of both tobacco leaf protoplasts and human peripheral blood mononuclear cells.

Science.gov (United States)

Goverse, A; Rouppe van der Voort, J; Roppe van der Voort, C; Kavelaars, A; Smant, G; Schots, A; Bakker, J; Helder, J

1999-10-01

Naturally induced secretions from infective juveniles of the potato cyst nematode Globodera rostochiensis co-stimulate the proliferation of tobacco leaf protoplasts in the presence of the synthetic phytohormones alpha-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). With the use of a protoplast-based bioassay, a low-molecular-weight peptide(s) (cyst nematode secretions also co-stimulated mitogenesis in human peripheral blood mononuclear cells (PBMC). The stimulation of plant cells isolated from nontarget tissue--these nematodes normally invade the roots of potato plants--suggests the activation of a general signal transduction mechanism(s) by an oligopeptide(s) secreted by the nematode. Whether a similar oligopeptide-induced mechanism underlies human PBMC activation remains to be investigated. Reactivation of the cell cycle is a crucial event in feeding cell formation by cyst nematodes. The secretion of a mitogenic low-molecular-weight peptide(s) by infective juveniles of the potato cyst nematode could contribute to the redifferentiation of plant cells into such a feeding cell.

The use of flow cytometry to monitor chitin synthesis in regenerating protoplasts of Candida albicans.

Science.gov (United States)

Hector, R F; Braun, P C; Hart, J T; Kamarck, M E

1990-01-01

Flow cytometry was used to monitor chitin synthesis in regenerating protoplasts of the yeast Candida albicans. Comparisons of cells stained with Calcofluor White, a fluorochrome with known affinity for chitin, and cells incubated in the presence of N-[3H]-acetylglucosamine, the precursor substrate for chitin, showed a linear relationship between fluorescence and incorporation of label over time. Changes in both the fluorescence and light scatter of regenerating protoplasts treated with inhibitors of fungal chitin synthase were also quantitated by flow cytometry.

Nitric oxide in guard cells as an important secondary messenger during stomatal closure

Directory of Open Access Journals (Sweden)

Gunja eGayatri

2013-10-01

Full Text Available he modulation of guard cell function is the basis of stomatal closure, essential for optimizing water use and CO2 uptake by leaves. Nitric oxide (NO in guard cells plays a very important role as a secondary messenger during stomatal closure induced by effectors, including hormones. For example, exposure to abscisic acid (ABA triggers a marked increase in NO of guard cells, well before stomatal closure. In guard cells of multiple species, like Arabidopsis, Vicia and pea, exposure to ABA or methyl jasmonate or even microbial elicitors (e.g. chitosan induces production of NO as well as reactive oxygen species (ROS. The role of NO in stomatal closure has been confirmed by using NO donors (e.g. SNP and NO scavengers (like cPTIO and inhibitors of NOS (L-NAME or NR (tungstate. Two enzymes: a L-NAME-sensitive, nitric oxide synthase (NOS-like enzyme and a tungstate-sensitive nitrate reductase (NR, can mediate ABA-induced NO rise in guard cells. However, the existence of true NOS in plant tissues and its role in guard cell NO-production are still a matter of intense debate. Guard cell signal transduction leading to stomatal closure involves the participation of several components, besides NO, such as cytosolic pH, ROS, free Ca2+ and phospholipids. Use of fluorescent dyes has revealed that the rise in NO of guard cells occurs after the increase in cytoplasmic pH and ROS. The rise in NO causes an elevation in cytosolic free Ca2+ and promotes the efflux of cations as well as anions from guard cells. Stomatal guard cells have become a model system to study the signalling cascade mechanisms in plants, particularly with NO as a dominant component. The interrelationships and interactions of NO with cytosolic pH, ROS, and free Ca2+ are quite complex and need further detailed examination. While assessing critically the available literature, the present review projects possible areas of further work related to NO-action in stomatal guard cells.

Actin filaments regulate the adhesion between the plasma membrane and the cell wall of tobacco guard cells.

Science.gov (United States)

Yu, Qin; Ren, Jing-Jing; Kong, Lan-Jing; Wang, Xiu-Ling

2018-01-01

During the opening and closing of stomata, guard cells undergo rapid and reversible changes in their volume and shape, which affects the adhesion of the plasma membrane (PM) to the cell wall (CW). The dynamics of actin filaments in guard cells are involved in stomatal movement by regulating structural changes and intracellular signaling. However, it is unclear whether actin dynamics regulate the adhesion of the PM to the CW. In this study, we investigated the relationship between actin dynamics and PM-CW adhesion by the hyperosmotic-induced plasmolysis of tobacco guard cells. We found that actin filaments in guard cells were depolymerized during mannitol-induced plasmolysis. The inhibition of actin dynamics by treatment with latrunculin B or jasplakinolide and the disruption of the adhesion between the PM and the CW by treatment with RGDS peptide (Arg-Gly-Asp-Ser) enhanced guard cell plasmolysis. However, treatment with latrunculin B alleviated the RGDS peptide-induced plasmolysis and endocytosis. Our results reveal that the actin depolymerization is involved in the regulation of the PW-CW adhesion during hyperosmotic-induced plasmolysis in tobacco guard cells.

Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells

KAUST Repository

Hu, Honghong; Boisson-Dernier, Auré lien; Israelsson-Nordströ m, Maria; Bö hmer, Maik; Xue, Shaowu; Ries, Amber; Godoski, Jan; Kuhn, Josef M.; Schroeder, Julian I.

2009-01-01

photosynthesis and can function in guard cells. Furthermore, guard cell betaca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian alphaCAII complements the reduced sensitivity of ca1 ca4 plants, showing

Interaction of E. coli DNA with tobacco mesophyll protoplasts

International Nuclear Information System (INIS)

Heyn, R.F.

1975-01-01

This chapter is part of a dissertation dealing with the interaction of DNA with protoplasts. Having established the length of time during which tobacco mesophyll protoplasts do not synthesize DNA following their isolation, it is important to know the extent of DNA uptake just before the onset of DNA synthesis (and possible integration) and to find optimal conditions for this uptake. Therefore, the association of E. coli DNA with tobacco protoplasts was studied. Care should be taken with the interpretation of ''uptake'' results: adsorption phenomena play a very important role and may do so at the plasmalemma of naked protoplasts. To solve the problems involved, the use of radiation-damaged DNA was attempted. With E. coli DNA possessing a large number of thymine containing pyrimidine dimers, the loss of dimers from DNA recovered from treated protoplasts was tested in order to obtain an indication of ''real'' uptake. The results are reported

A high efficiency technique for the generation of transgenic sugar beets from stomatal guard cells

NARCIS (Netherlands)

Hall, R.D.; Riksen-Bruinsma, T.; Weyens, G.; Rosquin, I.J.; Denys, R.N.; Evans, I.J.; Lathouwers, J.E.; LefObvre, M.P.; Dunwell, J.M.; Tunen, van A.; Krens, F.A.

1996-01-01

An optimized protocol has been developed for the efficient and rapid genetic modification of sugar beet (Beta vulgaris L). A polyethylene glycol- mediated DNA transformation technique could be applied to protoplast populations enriched specifically for a single totipotent cell type derived from

Common and unique elements of the ABA-regulated transcriptome of Arabidopsis guard cells

Directory of Open Access Journals (Sweden)

Zhao Zhixin

2011-05-01

Full Text Available Abstract Background In the presence of drought and other desiccating stresses, plants synthesize and redistribute the phytohormone abscisic acid (ABA. ABA promotes plant water conservation by acting on specialized cells in the leaf epidermis, guard cells, which border and regulate the apertures of stomatal pores through which transpirational water loss occurs. Following ABA exposure, solute uptake into guard cells is rapidly inhibited and solute loss is promoted, resulting in inhibition of stomatal opening and promotion of stomatal closure, with consequent plant water conservation. There is a wealth of information on the guard cell signaling mechanisms underlying these rapid ABA responses. To investigate ABA regulation of gene expression in guard cells in a systematic genome-wide manner, we analyzed data from global transcriptomes of guard cells generated with Affymetrix ATH1 microarrays, and compared these results to ABA regulation of gene expression in leaves and other tissues. Results The 1173 ABA-regulated genes of guard cells identified by our study share significant overlap with ABA-regulated genes of other tissues, and are associated with well-defined ABA-related promoter motifs such as ABREs and DREs. However, we also computationally identified a unique cis-acting motif, GTCGG, associated with ABA-induction of gene expression specifically in guard cells. In addition, approximately 300 genes showing ABA-regulation unique to this cell type were newly uncovered by our study. Within the ABA-regulated gene set of guard cells, we found that many of the genes known to encode ion transporters associated with stomatal opening are down-regulated by ABA, providing one mechanism for long-term maintenance of stomatal closure during drought. We also found examples of both negative and positive feedback in the transcriptional regulation by ABA of known ABA-signaling genes, particularly with regard to the PYR/PYL/RCAR class of soluble ABA receptors and

Common and unique elements of the ABA-regulated transcriptome of Arabidopsis guard cells

Science.gov (United States)

2011-01-01

Background In the presence of drought and other desiccating stresses, plants synthesize and redistribute the phytohormone abscisic acid (ABA). ABA promotes plant water conservation by acting on specialized cells in the leaf epidermis, guard cells, which border and regulate the apertures of stomatal pores through which transpirational water loss occurs. Following ABA exposure, solute uptake into guard cells is rapidly inhibited and solute loss is promoted, resulting in inhibition of stomatal opening and promotion of stomatal closure, with consequent plant water conservation. There is a wealth of information on the guard cell signaling mechanisms underlying these rapid ABA responses. To investigate ABA regulation of gene expression in guard cells in a systematic genome-wide manner, we analyzed data from global transcriptomes of guard cells generated with Affymetrix ATH1 microarrays, and compared these results to ABA regulation of gene expression in leaves and other tissues. Results The 1173 ABA-regulated genes of guard cells identified by our study share significant overlap with ABA-regulated genes of other tissues, and are associated with well-defined ABA-related promoter motifs such as ABREs and DREs. However, we also computationally identified a unique cis-acting motif, GTCGG, associated with ABA-induction of gene expression specifically in guard cells. In addition, approximately 300 genes showing ABA-regulation unique to this cell type were newly uncovered by our study. Within the ABA-regulated gene set of guard cells, we found that many of the genes known to encode ion transporters associated with stomatal opening are down-regulated by ABA, providing one mechanism for long-term maintenance of stomatal closure during drought. We also found examples of both negative and positive feedback in the transcriptional regulation by ABA of known ABA-signaling genes, particularly with regard to the PYR/PYL/RCAR class of soluble ABA receptors and their downstream targets

MAPK cascades in guard cell signal transduction

Directory of Open Access Journals (Sweden)

Yuree eLee

2016-02-01

Full Text Available Guard cells form stomata on the epidermis and continuously respond to endogenous and environmental stimuli to fine-tune the gas exchange and transpirational water loss, processes which involve mitogen-activated protein kinase (MAPK cascades. MAPKs form three-tiered kinase cascades with MAPK kinases and MAPK kinase kinases, by which signals are transduced to the target proteins. MAPK cascade genes are highly conserved in all eukaryotes, and they play crucial roles in myriad developmental and physiological processes. MAPK cascades function during biotic and abiotic stress responses by linking extracellular signals received by receptors to cytosolic events and gene expression. In this review, we highlight recent findings and insights into MAPK-mediated guard cell signaling, including the specificity of MAPK cascades and the remaining questions.

Infection of potato mesophyll protoplasts with five plant viruses.

Science.gov (United States)

Barker, H; Harrison, B D

1982-12-01

Methods are described for preparing potato mesophyll protoplasts that are suitable for infection with inocula of virus nucleoprotein or RNA. The protoplasts could be infected with four sap-transmissible viruses (tobacco mosaic, tobacco rattle, tobacco ringspot and tomato black ring viruses) and with potato leafroll virus, which is not saptransmissible. No differences were observed in ability to infect protoplasts with potato leafroll virus strains differing either in virulence in intact plants or in aphid transmissibility.

Proteins synthesized in tobacco mosaic virus infected protoplasts

NARCIS (Netherlands)

Huber, R.

1979-01-01

The study described here concerns the proteins, synthesized as a result of tobacco mosaic virus (TMV) multiplication in tobacco protoplasts and in cowpea protoplasts. The identification of proteins involved in the TMV infection, for instance in the virus RNA replication, helps to elucidate

Protoplast preparation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme

International Nuclear Information System (INIS)

Hassan Hamdani Mutaat; Mat Rasol Awang

2004-01-01

The objective of this study was to determine the optimum parameters of the factors influencing protoplast isolation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme from Trichoderma harzianurm. The study was conducted by manipulating the variables of the factors affecting protoplast isolation, such as age of mycelium culture, period for lysing of mycelium, concentration of lysing enzyme and concentration of osmotic stabilizer. The highest protoplast yield of 8.3 x 104 protoplast/ml was achieved when a 3-day P. sajor-caju mycelium, cultured statically, was incubated for 3 hours in a lytic mixture containing 7.5 mg/ml lysing enzyme and 1.2 M ammonium sulfate as osmotic stabilizer. This protoplast yield, however, is insufficient for regeneration and protoplast fusion works. (Author)

Poinsettia protoplasts - a simple, robust and efficient system for transient gene expression studies

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Pitzschke Andrea

2012-05-01

Full Text Available Abstract Background Transient gene expression systems are indispensable tools in molecular biology. Yet, their routine application is limited to few plant species often requiring substantial equipment and facilities. High chloroplast and chlorophyll content may further impede downstream applications of transformed cells from green plant tissue. Results Here, we describe a fast and simple technique for the high-yield isolation and efficient transformation (>70% of mesophyll-derived protoplasts from red leaves of the perennial plant Poinsettia (Euphorbia pulccherrima. In this method no particular growth facilities or expensive equipments are needed. Poinsettia protoplasts display an astonishing robustness and can be employed in a variety of commonly-used downstream applications, such as subcellular localisation (multi-colour fluorescence or promoter activity studies. Due to low abundance of chloroplasts or chromoplasts, problems encountered in other mesophyll-derived protoplast systems (particularly autofluorescence are alleviated. Furthermore, the transgene expression is detectable within 90 minutes of transformation and lasts for several days. Conclusions The simplicity of the isolation and transformation procedure renders Poinsettia protoplasts an attractive system for transient gene expression experiments, including multi-colour fluorescence, subcellular localisation and promoter activity studies. In addition, they offer hitherto unknown possibilities for anthocyan research and industrial applications.

Properties of Single K+ and Cl− Channels in Asclepias tuberosa Protoplasts 1

Science.gov (United States)

Schauf, Charles L.; Wilson, Kathryn J.

1987-01-01

Potassium and chloride channels were characterized in Asclepias tuberosa suspension cell derived protoplasts by patch voltage-clamp. Whole-cell currents and single channels in excised patches had linear instantaneous current-voltage relations, reversing at the Nernst potentials for K+ and Cl−, respectively. Whole cell K+ currents activated exponentially during step depolarizations, while voltage-dependent Cl− channels were activated by hyperpolarizations. Single K+ channel conductance was 40 ± 5 pS with a mean open time of 4.5 milliseconds at 100 millivolts. Potassium channels were blocked by Cs+ and tetraethylammonium, but were insensitive to 4-aminopyridine. Chloride channels had a single-channel conductance of 100 ± 17 picosiemens, mean open time of 8.8 milliseconds, and were blocked by Zn2+ and ethacrynic acid. Whole-cell Cl− currents were inhibited by abscisic acid, and were unaffected by indole-3-acetic acid and 2,4-dichlorophenoxyacetic acid. Since internal and external composition can be controlled, patch-clamped protoplasts are ideal systems for studying the role of ion channels in plant physiology and development. Images Fig. 5 PMID:16665712

Isolation and regeneration protoplast of an oil palm pathogen, Ganoderma boninense

Science.gov (United States)

Irene, Liza Isaac; Bakar, Farah Diba Abu; Idris, Abu Seman; Murad, Abdul Munir Abdul

2015-09-01

Ganoderma boninense is a known cause for basal stem rot (BSR) in oil palm. Thus, to curb the infection towards oil palm, the establishment of protoplast isolation and regeneration protocol is crucial to be studied. This will provide information on the functional genes especially those which leads towards infection and pathogenicity. In this study, a method was outlined to isolated protoplast in G. boninense by manipulating parameters such as mycelium age, concentration of lysing enzyme, and duration of mycelia incubation in lytic solution. The results shows that from 0.1 g of wet weight mycelia, the highest protoplast yield obtained was 5.5 × 108 protoplast/ml using 5th day old culture in a lytic mixture containing 2.0 % of lysing enzyme incubated for 4 hours at 30 °C with agitation of 80-100 rpm. The highest percentage of protoplast regeneration obtained from this study was 0.2 % using CYM medium supplemented with 0.6 M sorbitol. To date, this is the first report of protoplast isolation and regeneration for this phytopathogen.

Transient and stable expression of marker genes in cotransformed Petunia protoplasts in relation to X-ray and UV-irradiation

International Nuclear Information System (INIS)

Benediktsson, I.; Köhler, F.; Schieder, O.

1991-01-01

Irradiation of protoplasts with X-rays or ultraviolet light does not seem to influence the level of transient expression of foreign DNA in Petunia protoplasts, whereas the number of stably transformed colonies is significantly raised. This may indicate that irradiation influences integration and/or the expression of marker genes and does not result in enhanced uptake rates of plasmids into protoplasts and cell nuclei. Co-transformation with plasmids carrying a gene for kanamycin resistance (neomycin phosphotransferase II) and a gene for hygromycin resistance (hygromycin phosphotransferase) revealed that the cotransformation rates were not stimulated by irradiation when measuring expression

Plant regeneration from leaf protoplasts of Solanum torvum.

Science.gov (United States)

Guri, A; Volokita, M; Sink, K C

1987-07-01

A protocol to obtain regenerated plants from protoplasts of Solanum torvum Sw a wild species of eggplant resistant to Verticillium wilt is reported. Leaf protoplasts were enzymatically isolated from six-week old seedlings grown in a controlled environment chamber. Protoplasts were plated on modified KM medium (0.4 M glucose)+(mg/l): 1.0 p-chlorophenoxyacetic acid (CPA)+1.0 naphthaleneacetic acid (NAA)+0.5 6-benzylaminopurine (BAP) and 0.02 abscisic acid (ABA). The protoplast density was 5×10(4) per ml with 5 ml placed in each of two quadrants in X-dishes (100×15 mm). The reservoir medium was modified KM+(mg/l): 0.1 NAA+0.5 BAP+0.1 M sucrose+0.1 M mannitol+0.6% washed agar+1% activated charcoal. Dishes were initially placed in the dark at 27°C. Protoplast division was initiated in 1-2 weeks and 4 weeks later p-calli were 1-3 mm. Plating efficiency was 11% when measured at 3 weeks. Six-week old p-calli were transferred individually onto Whatman No. 1 filter paper layered on modified KM (0.15 M sucrose)+mg/l: 2.0 indoleacetic acid (IAA)+2.0 zeatin+0.5% washed agar for 2 weeks. Subsequently, shoots occurred within 4 weeks at 70% efficiency on MS+30 g/l sucrose+2 mg/l zeatin. Shoots were rooted on half strength MS+10 g/l sucrose.

Single guard cell recordings in intact plants : light-induced hyperpolarization of the plasma membrane

NARCIS (Netherlands)

Roelfsema, MRG; Steinmeyer, R; Staal, M; Hedrich, R

Guard cells are electrically isolated from other plant cells and therefore offer the unique possibility to conduct current- and voltage-clamp recordings on single cells in an intact plant. Guard cells in their natural environment were impaled with double-barreled electrodes and found to exhibit

Protoplast isolation, transient transformation of leaf mesophyll protoplasts and improved Agrobacterium-mediated leaf disc infiltration of Phaseolus vulgaris: tools for rapid gene expression analysis.

Science.gov (United States)

Nanjareddy, Kalpana; Arthikala, Manoj-Kumar; Blanco, Lourdes; Arellano, Elizabeth S; Lara, Miguel

2016-06-24

Phaseolus vulgaris is one of the most extensively studied model legumes in the world. The P. vulgaris genome sequence is available; therefore, the need for an efficient and rapid transformation system is more imperative than ever. The functional characterization of P. vulgaris genes is impeded chiefly due to the non-amenable nature of Phaseolus sp. to stable genetic transformation. Transient transformation systems are convenient and versatile alternatives for rapid gene functional characterization studies. Hence, the present work focuses on standardizing methodologies for protoplast isolation from multiple tissues and transient transformation protocols for rapid gene expression analysis in the recalcitrant grain legume P. vulgaris. Herein, we provide methodologies for the high-throughput isolation of leaf mesophyll-, flower petal-, hypocotyl-, root- and nodule-derived protoplasts from P. vulgaris. The highly efficient polyethylene glycol-mannitol magnesium (PEG-MMG)-mediated transformation of leaf mesophyll protoplasts was optimized using a GUS reporter gene. We used the P. vulgaris SNF1-related protein kinase 1 (PvSnRK1) gene as proof of concept to demonstrate rapid gene functional analysis. An RT-qPCR analysis of protoplasts that had been transformed with PvSnRK1-RNAi and PvSnRK1-OE vectors showed the significant downregulation and ectopic constitutive expression (overexpression), respectively, of the PvSnRK1 transcript. We also demonstrated an improved transient transformation approach, sonication-assisted Agrobacterium-mediated transformation (SAAT), for the leaf disc infiltration of P. vulgaris. Interestingly, this method resulted in a 90 % transformation efficiency and transformed 60-85 % of the cells in a given area of the leaf surface. The constitutive expression of YFP further confirmed the amenability of the system to gene functional characterization studies. We present simple and efficient methodologies for protoplast isolation from multiple P

DOF-binding sites additively contribute to guard cell-specificity of AtMYB60 promoter

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Cominelli Eleonora

2011-11-01

Full Text Available Abstract Background We previously demonstrated that the Arabidopsis thaliana AtMYB60 protein is an R2R3MYB transcription factor required for stomatal opening. AtMYB60 is specifically expressed in guard cells and down-regulated at the transcriptional levels by the phytohormone ABA. Results To investigate the molecular mechanisms governing AtMYB60 expression, its promoter was dissected through deletion and mutagenesis analyses. By studying different versions of AtMYB60 promoter::GUS reporter fusions in transgenic plants we were able to demonstrate a modular organization for the AtMYB60 promoter. Particularly we defined: a minimal promoter sufficient to confer guard cell-specific activity to the reporter gene; the distinct roles of different DOF-binding sites organised in a cluster in the minimal promoter in determining guard cell-specific expression; the promoter regions responsible for the enhancement of activity in guard cells; a promoter region responsible for the negative transcriptional regulation by ABA. Moreover from the analysis of single and multiple mutants we could rule out the involvement of a group of DOF proteins, known as CDFs, already characterised for their involvement in flowering time, in the regulation of AtMYB60 expression. Conclusions These findings shed light on the regulation of gene expression in guard cells and provide new promoter modules as useful tools for manipulating gene expression in guard cells, both for physiological studies and future biotechnological applications.

Genetic transformation of the white-rot fungus Dichomitus squalens using a new commercial protoplasting cocktail.

Science.gov (United States)

Daly, Paul; Slaghek, Gillian G; Casado López, Sara; Wiebenga, Ad; Hilden, Kristiina S; de Vries, Ronald P; Mäkelä, Miia R

2017-12-01

D. squalens, a white-rot fungus that efficiently degrades lignocellulose in nature, can be used in various biotechnological applications and has several strains with sequenced and annotated genomes. Here we present a method for the transformation of this basidiomycete fungus, using a recently introduced commercial ascomycete protoplasting enzyme cocktail, Protoplast F. In protoplasting of D. squalens mycelia, Protoplast F outperformed two other cocktails while releasing similar amounts of protoplasts to a third cocktail. The protoplasts released using Protoplast F had a regeneration rate of 12.5% (±6 SE). Using Protoplast F, the D. squalens monokaryon CBS464.89 was conferred with resistance to the antibiotics hygromycin and G418 via polyethylene glycol mediated protoplast transformation with resistance cassettes expressing the hygromycin phosphotransferase (hph) and neomycin phosphotransferase (nptII) genes, respectively. The hph gene was expressed in D. squalens using heterologous promoters from genes encoding β-tubulin or glyceraldehyde 3-phosphate dehydrogenase. A Southern blot confirmed integration of a resistance cassette into the D. squalens genome. An average of six transformants (±2 SE) were obtained when at least several million protoplasts were used (a transformation efficiency of 0.8 (±0.3 SE) transformants per μg DNA). Transformation of D. squalens demonstrates the suitability of the Protoplast F cocktail for basidiomycete transformation and furthermore can facilitate understanding of basidiomycete gene function and development of improved strains for biotechnological applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis1[OPEN

Science.gov (United States)

Rui, Yue; Anderson, Charles T.

2016-01-01

Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3je5 mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface. PMID:26729799

Protoplasting impact on polyketide activity and characterization of the interspecific fusants from Streptomyces spp

International Nuclear Information System (INIS)

Slama, N.; Lazim, H.; Barkallah, Insaf; Abbassi, M.; Ben Hassen, A.; Limam, F.

2009-01-01

Streptomycetes are gram-positive, soil-inhabiting bacteria of the order Actinomycetales. These organisms exhibit an unusual, developmentally complex life cycle and produce many economically important secondary metabolites, such as antibiotics, immunosuppressants, insecticides, and antitumor agents. Streptomyces species have been the subject of genetic investigation for over 50 years, with many studies focusing on the production of bioactives compounds. The protoplast formation and regeneration are important processes, and they are a major step following genetic manipulations such as fusion and DNA-mediated transformation, which can improve antibiotic production. The protoplast fusion, transformation and improved fermentation features can be used to regenerate strains with increased antibiotic activity. Local Streptomyces spp. CN207 produce a broad range of secondary metabolites which is active against bacteria and fungi. This strain was used as a donor and S. coelicolor strain M145 was used as a recipient host for protoplast fusion. The protoplast fusion resulted in increased isolation of variants with higher antibiotic activity. Recombinant Streptomyces coelicolor PF04 was increased 10 times more than the wild strain. The antimicrobial activity from PF04 strain was studied using the disc method agar. TLC analysis confirmed that the Rf of cell extract for PF04 strain is identical to antimicrobial compound of Streptomyces CN207. Our results confirm the possibility of transferring antibiotics cluster genes by fusion. In fact, many of the selective markers such as Ticarcillin, Cefalotin, Oxacillin and Cefotaxim were transferred during the protoplast fusion. PFGE analysis and DNA-hybridization confirmed the presence of homologous fragments between a wild-type Streptomyces CN207 and a recombinant S. coelicolor PF04

Patterns of indole alkaloids synthesis in response to heat shock, 5-azacytidine and Na-butyrate treatment of cultured catharanthus roseus mesophyll protoplasts

International Nuclear Information System (INIS)

Saleem, M.; Cutler, A.J.

1986-01-01

Alkaloids of C. roseus are in high demand for therapeutic and other reasons. Cultured Catharanthus cells can produce limited quantities of these alkaloids. The authors have found that cultured mesophyll protoplasts in the presence of 14 C-Tryptamine are capable of synthesizing alkaloids. The pattern of alkaloids synthesis changes when protoplasts are subjected to a heat shock at 37 0 C. The heat shocked protoplasts incorporated 33% more 14 C-Tryptamine and produced 3 new types of alkaloids. Treatment of protoplasts with 5-azacytidine, a DNA hypomethylating agent and Na-butyrate which induces hyperacetylation of histones produced qualitative and quantitative changes in the alkaloid pattern. Four new alkaloids following the above treatments were detected by TLC and HPLC of the extracts. It is suggested that the alkaloid pattern of the cultured protoplasts can be altered by treatment with compounds known as regulators of gene expression. Work is in progress to isolate and identify these new alkaloids

Guard Cell Signal Transduction Network: Advances in Understanding Abscisic Acid, CO2, and Ca2+ Signaling

KAUST Repository

Kim, Tae-Houn

2010-05-04

Stomatal pores are formed by pairs of specialized epidermal guard cells and serve as major gateways for both CO2 influx into plants from the atmosphere and transpirational water loss of plants. Because they regulate stomatal pore apertures via integration of both endogenous hormonal stimuli and environmental signals, guard cells have been highly developed as a model system to dissect the dynamics and mechanisms of plant-cell signaling. The stress hormone ABA and elevated levels of CO2 activate complex signaling pathways in guard cells that are mediated by kinases/phosphatases, secondary messengers, and ion channel regulation. Recent research in guard cells has led to a new hypothesis for how plants achieve specificity in intracellular calcium signaling: CO2 and ABA enhance (prime) the calcium sensitivity of downstream calcium-signaling mechanisms. Recent progress in identification of early stomatal signaling components are reviewed here, including ABA receptors and CO2-binding response proteins, as well as systems approaches that advance our understanding of guard cell-signaling mechanisms.

Guard Cell Signal Transduction Network: Advances in Understanding Abscisic Acid, CO2, and Ca2+ Signaling

KAUST Repository

Kim, Tae-Houn; Bö hmer, Maik; Hu, Honghong; Nishimura, Noriyuki; Schroeder, Julian I.

2010-01-01

Stomatal pores are formed by pairs of specialized epidermal guard cells and serve as major gateways for both CO2 influx into plants from the atmosphere and transpirational water loss of plants. Because they regulate stomatal pore apertures via integration of both endogenous hormonal stimuli and environmental signals, guard cells have been highly developed as a model system to dissect the dynamics and mechanisms of plant-cell signaling. The stress hormone ABA and elevated levels of CO2 activate complex signaling pathways in guard cells that are mediated by kinases/phosphatases, secondary messengers, and ion channel regulation. Recent research in guard cells has led to a new hypothesis for how plants achieve specificity in intracellular calcium signaling: CO2 and ABA enhance (prime) the calcium sensitivity of downstream calcium-signaling mechanisms. Recent progress in identification of early stomatal signaling components are reviewed here, including ABA receptors and CO2-binding response proteins, as well as systems approaches that advance our understanding of guard cell-signaling mechanisms.

Longevity of guard cell chloroplasts in falling leaves: implication for stomatal function and cellular aging.

Science.gov (United States)

Zeiger, E; Schwartz, A

1982-11-12

Guard cell chloroplasts in senescing leaves from 12 species of perennial trees and three species of annual plants survived considerably longer than their mesophyll counterparts. In Ginkgo biloba, stomata from yellow leaves opened during the day and closed at night; guard cell chloroplasts from these leaves showed fluorescence transients associated with electron transport and photophosphorylation. These findings indicate that guard cell chloroplasts are highly conserved throughout the life-span of the leaf and that leaves retain stomatal control during senescence.

Evidence for some signal transduction elements involved in UV-light-dependent responses in parsley protoplasts

International Nuclear Information System (INIS)

Frohnmeyer, H.; Bowler, C.; Schäfer, E.

1997-01-01

The signalling pathways used by UV-light are largely unknown. Using protoplasts from a heterotrophic parsley (Petroselinum crispum L.) cell culture that exclusively respond to UV-B light between 300 and 350 nm with a fast induction of genes encoding flavonoid biosynthetic enzymes, information was obtained about the UV-light signal transduction pathway for chalcone synthase (CHS) and phenylalanine ammonia-lyase (PAL) gene expression. Pharmacological effectors which influence intracellular calcium levels, calmodulin and the activity of serine/threonine kinases also changed the UV-light-dependent expression of these genes. This evaluation indicated the participation of these components on the UV-B-mediated signal transduction cascade to CHS. In contrast, neither membrane-permeable cyclic GMP nor the tyrosine kinase inhibitor genistein affected CHS or PAL expression. Similar results were obtained in protoplasts, which have been transiently transformed with CHS-promoter/GUS (β-glucuronidase) reporter fusion constructs. The involvement of calcium and calmodulin was further indicated in a cell-free light-responsive in vitro transcription system from evacuolated parsley protoplasts. In conclusion, there is evidence now that components of the UV-light-dependent pathway leading to the CHS-promoter are different from the previously characterized cGMP-dependent pathway to CHS utilized by phytochrome in soybean (Glycine max) and tomato seedlings (Lycopersicon esculentum). (author)

Influences of explant type and enzyme incubation on isolated protoplast density and viability in two garlic cultivars

International Nuclear Information System (INIS)

Metwally, E.I.

2014-01-01

The present study reports on optimizing protoplast isolation and fusion in two garlic cultivars Balady and Seds 40. Protoplast density and viability were investigated in four different explants (etiolated and green parts of the pseudostem and lower and upper parts of the leaves) under enzyme incubation for 1, 2, 3 and 4 h. Among different explants, used for protoplast isolation in Balady cultivar, the upper and lower parts of the leaves produced the highest number of total protoplasts (70 and 66 pps/0.1 ml) at 4 and 3 h enzyme incubation, respectively. However, the etiolated part of pseudostem produced the highest number of viable protoplast in which 52.5 pps/0.1 ml were obtained at 3 h enzyme incubation. For protoplast isolation in Seds 40 cultivar, the highest number of total protoplasts (125 and 107.5 pps/0.1 ml) as well as viable protoplasts (105 and 107.5 pps/0.1 ml) was obtained from the etiolated and the green parts of pseudostem, respectively. The cultivar Seds 40 yielded higher total and viable protoplasts than Balady cultivar. Isolated protoplasts of Seds 40 and Balady were fused successfully at a protoplast density of 1 * 105 using either physical and/or electrical method. Optimization of the source of plant material as well as protoplast isolation conditions for garlic is a crucial step towards a successful protoplast fusion and subsequent colony formation. (author)

Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast-derived cells.

Science.gov (United States)

Houba-Hérin, N; Domin, M; Pédron, J

1994-07-01

Nicotiana plumbaginifolia haploid protoplasts were co-transformed with two plasmids, one with a NPT-II/Ds element and one with a gene encoding an amino-terminal truncated Ac transposase. It is shown that Ds can efficiently transpose from extrachromosomal DNA to N. plumbaginifolia chromosomes when the Ac transposase gene is present in trans. Ds has been shown to have transposed into the plant genome in a limited number of copies (1.9 copies per genome), for 21/32 transgenic lines tested. The flanking sequences present in the original plasmid are missing in these 21 plants. In only two of 21 plants was part of the transposase construct integrated. By segregation analysis of transgenic progeny, Ds was shown to be present in the heterozygous state in 10 lines even though haploid protoplasts had been originally transformed. This observation could indicate that integration occurred after or during DNA replication that leads to protoplast diploidization.

Can protoplast production from in vitro cultured shoots of Tanacetum vary during the season?

Directory of Open Access Journals (Sweden)

M. KESKITALO

2008-12-01

Full Text Available Two different experiments were carried out to study the production of protoplasts and the variation of protoplast yield from in vitro cultured shoot tips of tansy (Tanacetum vulgare L. and pyrethrum (Tanacetum cinerariifolium (Trevir. Schiltz-Bip. In the first experiment, light had more pronouced effect for tansy than for pyrethrum. When the donor tissues of tansy were cultured under high light intensity the leaves contained anthocyanin and became brown during enzyme maceration. In contrast, donor tissues cultured under low light intensity produced leaves without anthocyanin. Depending on the light intensity of donor tissues, on average 5.8 - 6.8 x 106 and 3.4 - 4.3 x 106 protoplasts were isolated from one gram of mesophyll leaves of tansy and pyrethrum, respectively. In the second experiment, the production of protoplasts from tansy and pyrethrum varied seasonally. The most successful season for the production of protoplasts from in vitro cultured shoot tips was between December and April, when also the highest number of protoplasts could be isolated. It was not possible to state whether Tanacetum species have rhythms, which could cause physiological or chemical changes for the in vitro grown shoot tips. However, some external or internal, possible seasonal-dependent stimuli may have caused variation in the number of protoplasts isolated from tansy and pyrethrum and favoured protoplast production during winter and spring.

Analysis of the effects of cerium on calcium ion in the protoplasts of ...

African Journals Online (AJOL)

The laser-scanning confocal microscopy has become a routine technique and indispensable tool for cell biological studies. In this study, the probe Fluo-3 AM was used to research the instantaneous changes of calcium ion (Ca2+) in the protoplasts of Arabidopsis thaliana. The laser-scanning mode of confocal microscope is ...

Rice Stomatal Closure Requires Guard Cell Plasma Membrane ATP-Binding Cassette Transporter RCN1/OsABCG5.

Science.gov (United States)

Matsuda, Shuichi; Takano, Sho; Sato, Moeko; Furukawa, Kaoru; Nagasawa, Hidetaka; Yoshikawa, Shoko; Kasuga, Jun; Tokuji, Yoshihiko; Yazaki, Kazufumi; Nakazono, Mikio; Takamure, Itsuro; Kato, Kiyoaki

2016-03-07

Water stress is one of the major environmental stresses that affect agricultural production worldwide. Water loss from plants occurs primarily through stomatal pores. Here, we report that an Oryza sativa half-size ATP-binding cassette (ABC) subfamily G protein, RCN1/OsABCG5, is involved in stomatal closure mediated by phytohormone abscisic acid (ABA) accumulation in guard cells. We found that the GFP-RCN1/OsABCG5-fusion protein was localized at the plasma membrane in guard cells. The percentage of guard cell pairs containing both ABA and GFP-RCN1/OsABCG5 increased after exogenous ABA treatment, whereas they were co-localized in guard cell pairs regardless of whether exogenous ABA was applied. ABA application resulted in a smaller increase in the percentage of guard cell pairs containing ABA in rcn1 mutant (A684P) and RCN1-RNAi than in wild-type plants. Furthermore, polyethylene glycol (drought stress)-inducible ABA accumulation in guard cells did not occur in rcn1 mutants. Stomata closure mediated by exogenous ABA application was strongly reduced in rcn1 mutants. Finally, rcn1 mutant plants had more rapid water loss from detached leaves than the wild-type plants. These results indicate that in response to drought stress, RCN1/OsABCG5 is involved in accumulation of ABA in guard cells, which is indispensable for stomatal closure. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

14C fixation by leaves and leaf cell protoplasts of the submerged aquatic angiosperm Potamogeton lucens: Carbon dioxide or bicarbonate?

International Nuclear Information System (INIS)

Staal, M.; Elzenga, J.T.M.; Prins, H.B.A.

1989-01-01

Protoplasts were isolated from leaves of the aquatic angiosperm Potamogeton lucens L. The leaves utilize bicarbonate as a carbon source for photosynthesis, and show polarity; that is acidification of the periplasmic space of the lower, and alkalinization of the space near the upper leaf side. At present there are two models under consideration for this photosynthetic bicarbonate utilization process: conversion of bicarbonate into free carbon dioxide as a result of acidification and, second, a bicarbonate-proton symport across the plasma membrane. Carbon fixation of protoplasts was studied at different pH values and compared with that in leaf strips. Using the isotopic disequilibrium technique, it was established that carbon dioxide and not bicarbonate was the form in which DIC actually crossed the plasma membrane. It is concluded that there is probably no true bicarbonate transport system at the plasma membrane of these cells and that bicarbonate utilization in this species apparently rests on the conversion of bicarbonate into carbon dioxide. Experiments with acetazolamide, an inhibitor of periplasmic carbonic anhydrase, and direct measurements of carbonic anhydrase activity in intact leaves indicate that in this species the role of this enzyme for periplasmic conversion of bicarbonate into carbon dioxide is insignificant

Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum.

Science.gov (United States)

Ju, Jong Il; Ko, Jung-Moon; Kim, So Hyeon; Baek, Ju Yeoul; Cha, Hyeon-Cheol; Lee, Sang Hoon

2006-08-01

In plant cell culture, the delivery of nutrition and gas (mainly oxygen) to the cells is the most important factor for viability. In this paper, we propose a polydimethylsiloxane (PDMS)-based microculture system that is designed to have good aeration. PDMS is known to have excellent air permeability, and through the experimental method, we investigated the relation between the degree of air delivery and the thickness of the PDMS sheet covering the culture chamber. We determined the proper thickness of the cover sheet, and cultured protoplasts of Nicotiana tabacum in a culture chamber covered with a PDMS sheet having thickness of 400 microm. The cells were successfully divided, and lived well inside the culture chamber for 10 days. In addition, protoplasts were cultured inside the culture chambers covered with the cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.

[Isolation and regeneration of the protoplasts of the streptomycete producers of actinomycins C and X].

Science.gov (United States)

Orlova, T I; Masha, G G; Kliueva, N A

1986-09-01

Protoplasts of S. michiganensis, S. chrysomallus and Streptomyces sp. 26-115, organisms producing actinomycins C and X form in hypertonic salt solution under the action of 3-4,5 mg/ml of lysozyme on the mycelium suspension. For protoplasting, the streptomycetes were grown on the soybean medium in the presence of 0.2-0.8 per cent of glycine. The mycelium of the streptomycete exponential growth phase was more favourable for protoplast formation. Protoplast regeneration was studied on the medium described by Okanishi et al. The quantitative composition of this medium was not optimal for regeneration of protoplasts of the above streptomycetes. The level of their regeneration depended to various extents on concentration of phosphate, magnesium and calcium ions and sucrose in the regeneration medium.

A computational approach for inferring the cell wall properties that govern guard cell dynamics.

Science.gov (United States)

Woolfenden, Hugh C; Bourdais, Gildas; Kopischke, Michaela; Miedes, Eva; Molina, Antonio; Robatzek, Silke; Morris, Richard J

2017-10-01

Guard cells dynamically adjust their shape in order to regulate photosynthetic gas exchange, respiration rates and defend against pathogen entry. Cell shape changes are determined by the interplay of cell wall material properties and turgor pressure. To investigate this relationship between turgor pressure, cell wall properties and cell shape, we focused on kidney-shaped stomata and developed a biomechanical model of a guard cell pair. Treating the cell wall as a composite of the pectin-rich cell wall matrix embedded with cellulose microfibrils, we show that strong, circumferentially oriented fibres are critical for opening. We find that the opening dynamics are dictated by the mechanical stress response of the cell wall matrix, and as the turgor rises, the pectinaceous matrix stiffens. We validate these predictions with stomatal opening experiments in selected Arabidopsis cell wall mutants. Thus, using a computational framework that combines a 3D biomechanical model with parameter optimization, we demonstrate how to exploit subtle shape changes to infer cell wall material properties. Our findings reveal that proper stomatal dynamics are built on two key properties of the cell wall, namely anisotropy in the form of hoop reinforcement and strain stiffening. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd and Society for Experimental Biology.

Expression of Arabidopsis Hexokinase in Citrus Guard Cells Controls Stomatal Aperture and Reduces Transpiration.

Science.gov (United States)

Lugassi, Nitsan; Kelly, Gilor; Fidel, Lena; Yaniv, Yossi; Attia, Ziv; Levi, Asher; Alchanatis, Victor; Moshelion, Menachem; Raveh, Eran; Carmi, Nir; Granot, David

2015-01-01

Hexokinase (HXK) is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1) under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species.

Transfer of herpes simplex virus thymidine kinase synthesized in bacteria by a high-expression plasmid to tissue culture cells by protoplast fusion

International Nuclear Information System (INIS)

Waldman, A.S.; Milman, G.

1984-01-01

The introduction of a protein into living tissue culture cells may permit the in vivo study of functions of the protein. The authors have previously described a high-efficiency-expression plasmid, pHETK2, containing the herpes simplex virus type 1 thymidine kinase (TK) gene which, upon temperature induction, causes TK to be synthesized as greater than 4% of the bacterial protein. In this report it is shown that enzymatically active TK was transferred to mouse Ltk- cells by polyethylene glycol-mediated fusion with protoplasts prepared from bacteria containing induced levels of TK. The presence of TK in the Ltk- cells was detected by the incorporation of [ 3 H]thymidine into cell nuclei as measured by autoradiography

Optimization of Production Conditions for Protoplasts and Polyethylene Glycol-Mediated Transformation of Gaeumannomyces tritici.

Science.gov (United States)

Wang, Mei; Zhang, Jie; Wang, Lanying; Han, Lirong; Zhang, Xing; Feng, Juntao

2018-05-24

Take-all, caused by Gaeumannomyces tritici , is one of the most important wheat root diseases worldwide, as it results in serious yield losses. In this study, G. tritici was transformed to express the hygromycin B phosphotransferase using a combined protoplast and polyethylene glycol (PEG)-mediated transformation technique. Based on a series of single-factor experimental results, three major factors-temperature, enzyme lysis time, and concentration of the lysing enzyme-were selected as the independent variables, which were optimized using the response surface methodology. A higher protoplast yield of 9.83 × 10⁷ protoplasts/mL was observed, and the protoplast vitality was also high, reaching 96.27% after optimization. Protoplasts were isolated under the optimal conditions, with the highest transformation frequency (46⁻54 transformants/μg DNA). Polymerase chain reaction and Southern blotting detection indicated that the genes of hygromycin phosphotransferase were successfully inserted into the genome of G. tritici . An optimised PEG-mediated protoplast transformation system for G. tritici was established. The techniques and procedures described will lay the foundation for establishing a good mutation library of G. tritici and could be used to transform other fungi.

Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill. protoplast yield

Directory of Open Access Journals (Sweden)

Elżbieta Kuźniak

2013-12-01

Full Text Available The effects of soil or in vitro grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 107/g FW was obtained from leaves of in vitro grown plants. Low light intensity during donor plants in vitro culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to transfer to enzyme mixture increased 4-fold the number of isolated protoplasts. Glycine and bovine serum albumin in the isolation medium did not significantly influence the protoplast yield.

Aquaporins Contribute to ABA-Triggered Stomatal Closure through OST1-Mediated Phosphorylation

Science.gov (United States)

Grondin, Alexandre; Rodrigues, Olivier; Verdoucq, Lionel; Merlot, Sylvain; Leonhardt, Nathalie; Maurel, Christophe

2015-01-01

Stomatal movements in response to environmental stimuli critically control the plant water status. Although these movements are governed by osmotically driven changes in guard cell volume, the role of membrane water channels (aquaporins) has remained hypothetical. Assays in epidermal peels showed that knockout Arabidopsis thaliana plants lacking the Plasma membrane Intrinsic Protein 2;1 (PIP2;1) aquaporin have a defect in stomatal closure, specifically in response to abscisic acid (ABA). ABA induced a 2-fold increase in osmotic water permeability (Pf) of guard cell protoplasts and an accumulation of reactive oxygen species in guard cells, which were both abrogated in pip2;1 plants. Open stomata 1 (OST1)/Snf1-related protein kinase 2.6 (SnRK2.6), a protein kinase involved in guard cell ABA signaling, was able to phosphorylate a cytosolic PIP2;1 peptide at Ser-121. OST1 enhanced PIP2;1 water transport activity when coexpressed in Xenopus laevis oocytes. Upon expression in pip2;1 plants, a phosphomimetic form (Ser121Asp) but not a phosphodeficient form (Ser121Ala) of PIP2;1 constitutively enhanced the Pf of guard cell protoplasts while suppressing its ABA-dependent activation and was able to restore ABA-dependent stomatal closure in pip2;1. This work supports a model whereby ABA-triggered stomatal closure requires an increase in guard cell permeability to water and possibly hydrogen peroxide, through OST1-dependent phosphorylation of PIP2;1 at Ser-121. PMID:26163575

Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells

KAUST Repository

Hu, Honghong

2009-12-13

The continuing rise in atmospheric CO2 causes stomatal pores in leaves to close and thus globally affects CO2 influx into plants, water use efficiency and leaf heat stress. However, the CO2-binding proteins that control this response remain unknown. Moreover, which cell type responds to CO2, mesophyll or guard cells, and whether photosynthesis mediates this response are matters of debate. We demonstrate that Arabidopsis thaliana double-mutant plants in the beta-carbonic anhydrases betaCA1 and betaCA4 show impaired CO2-regulation of stomatal movements and increased stomatal density, but retain functional abscisic-acid and blue-light responses. betaCA-mediated CO2-triggered stomatal movements are not, in first-order, linked to whole leaf photosynthesis and can function in guard cells. Furthermore, guard cell betaca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian alphaCAII complements the reduced sensitivity of ca1 ca4 plants, showing that carbonic anhydrase-mediated catalysis is an important mechanism for betaCA-mediated CO2-induced stomatal closure and patch clamp analyses indicate that CO2/HCO3- transfers the signal to anion channel regulation. These findings, together with ht1-2 (ref. 9) epistasis analysis demonstrate that carbonic anhydrases function early in the CO2 signalling pathway, which controls gas-exchange between plants and the atmosphere.

Expression of Arabidopsis hexokinase in citrus guard cells controls stomatal aperture and reduces transpiration

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Nitsan eLugassi

2015-12-01

Full Text Available Hexokinase (HXK is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1 under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species.

Genetic variability in regenerated Metarhizium flavoviride protoplasts

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Júlia Kuklinsky-Sobral

2004-03-01

Full Text Available Protoplast isolation and regeneration were evaluated in two wild-type and two colour mutant strains of Metarhizium flavoviride. Cultivation in liquid medium, followed by mycelium treatment with Novozym 234 in the presence of KCl 0.7M as osmotic stabilizer, produced 5.05 x 10(6 to 1.15 x 10(7x mL-1 protoplasts. The percentage of regeneration ranged from 6.65 to 27.92%. Following protoplast regeneration, one strain produced spontaneously stable morphological variant colonies. Although colonies with altered morphology have been reported in bacteria following protoplast regeneration, this is the first time that the same is described in a filamentous fungus. The original strain and one derived variant were tested for sensitivity to the fungicides benomyl and captan.A formação e regeneração de protoplastos foram avaliadas em duas linhagens selvagens e duas linhagens mutantes para coloração de conídios em Metarhizium flavoviride. O cultivo em meio líquido seguido do tratamento do micélio com Novozym 234 na presença de KCl 0,7 M como estabilizador osmótico, resultou na produção de 5,05´10(6 a 1,15´10(7 protoplastos´mL-1. A porcentagem de regeneração das diferentes linhagens variou de 6,65 a 27,92%. Após a regeneração, uma das linhagens selvagens produziu espontaneamente variantes estáveis, com morfologia alterada. Embora variantes morfológicos já tenham sido observados após regeneração de protoplastos em bactérias, esta parece ser a primeira vez que tal ocorrência é descrita em fungos filamentosos. Um desses variantes, além da linhagem selvagem da qual ele foi originado, foi testado para sensibilidade aos fungicidas benomil e captano.

Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing.

Science.gov (United States)

Xu, Xiaoyong; Xu, Xiaoling; Zhou, Yipeng; Zeng, Shaohua; Kong, Weiwen

2017-01-01

Protoplast isolation is a stress-inducing process, during which a variety of physiological and molecular alterations take place. Such stress response affects the expression of totipotency of cultured protoplasts. MicroRNAs (miRNAs) play important roles in plant growth, development and stress responses. However, the underlying mechanism of miRNAs involved in the protoplast totipotency remains unclear. In this study, high-throughput sequencing technology was used to sequence two populations of small RNA from calli and callus-derived protoplasts in Citrus reticulata Blanco. A total of 67 known miRNAs from 35 families and 277 novel miRNAs were identified. Among these miRNAs, 18 known miRNAs and 64 novel miRNAs were identified by differentially expressed miRNAs (DEMs) analysis. The expression patterns of the eight DEMs were verified by qRT-PCR. Target prediction showed most targets of the miRNAs were transcription factors. The expression levels of half targets showed a negative correlation to those of the miRNAs. Furthermore, the physiological analysis showed high levels of antioxidant activities in isolated protoplasts. In short, our results indicated that miRNAs may play important roles in protoplast-isolation response.

Improved efficiency of plant regeneration from protoplasts of eggplant Solanum melongena L.

Science.gov (United States)

Guri, A; Izhar, S

1984-12-01

Eggplant (Solanum melongena L.) mesophyll protoplasts were obtained from in vitro growing plants of line 410 and cv. 'Classic'. Relatively high (15%) plating efficiency was achieved using petri dishes with alternate quadrants containing reservoir medium (R medium + 1% activated charcoal) and culture medium. Shoot regeneration occurred within 6 weeks following initiation of protoplast culture.

Microarray Expression Analyses of Arabidopsis Guard Cells and Isolation of a Recessive Abscisic Acid Hypersensitive Protein Phosphatase 2C MutantW⃞

Science.gov (United States)

Leonhardt, Nathalie; Kwak, June M.; Robert, Nadia; Waner, David; Leonhardt, Guillaume; Schroeder, Julian I.

2004-01-01

Oligomer-based DNA Affymetrix GeneChips representing about one-third of Arabidopsis (Arabidopsis thaliana) genes were used to profile global gene expression in a single cell type, guard cells, identifying 1309 guard cell–expressed genes. Highly pure preparations of guard cells and mesophyll cells were isolated in the presence of transcription inhibitors that prevented induction of stress-inducible genes during cell isolation procedures. Guard cell expression profiles were compared with those of mesophyll cells, resulting in identification of 64 transcripts expressed preferentially in guard cells. Many large gene families and gene duplications are known to exist in the Arabidopsis genome, giving rise to redundancies that greatly hamper conventional genetic and functional genomic analyses. The presented genomic scale analysis identifies redundant expression of specific isoforms belonging to large gene families at the single cell level, which provides a powerful tool for functional genomic characterization of the many signaling pathways that function in guard cells. Reverse transcription–PCR of 29 genes confirmed the reliability of GeneChip results. Statistical analyses of promoter regions of abscisic acid (ABA)–regulated genes reveal an overrepresented ABA responsive motif, which is the known ABA response element. Interestingly, expression profiling reveals ABA modulation of many known guard cell ABA signaling components at the transcript level. We further identified a highly ABA-induced protein phosphatase 2C transcript, AtP2C-HA, in guard cells. A T-DNA disruption mutation in AtP2C-HA confers ABA-hypersensitive regulation of stomatal closing and seed germination. The presented data provide a basis for cell type–specific genomic scale analyses of gene function. PMID:14973164

Real-Time In Vivo Monitoring of Reactive Oxygen Species in Guard Cells.

Science.gov (United States)

Park, Ky Young; Roubelakis-Angelakis, Kalliopi A

2018-01-01

The intra-/intercellular homeostasis of reactive oxygen species (ROS), and especially of superoxides (O 2 .- ) and hydrogen peroxide (O 2 .- ) participate in signalling cascades which dictate developmental processes and reactions to biotic/abiotic stresses. Polyamine oxidases terminally oxidize/back convert polyamines generating H 2 O 2 . Recently, an NADPH-oxidase/Polyamine oxidase feedback loop was identified to control oxidative burst under salinity. Thus, the real-time localization/monitoring of ROS in specific cells, such as the guard cells, can be of great interest. Here we present a detailed description of the real-time in vivo monitoring of ROS in the guard cells using H 2 O 2 - and O 2 .- specific fluorescing probes, which can be used for studying ROS accumulation generated from any source, including the amine oxidases-dependent pathway, during development and stress.

Guard cell photosynthesis is critical for stomatal turgor production, yet does not directly mediate CO2 - and ABA-induced stomatal closing.

Science.gov (United States)

Azoulay-Shemer, Tamar; Palomares, Axxell; Bagheri, Andisheh; Israelsson-Nordstrom, Maria; Engineer, Cawas B; Bargmann, Bastiaan O R; Stephan, Aaron B; Schroeder, Julian I

2015-08-01

Stomata mediate gas exchange between the inter-cellular spaces of leaves and the atmosphere. CO2 levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO2 ]. [CO2 ] in leaves mediates stomatal movements. The role of guard cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll-deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard cell specific enhancer trap line. Our data show that more than 90% of guard cells were chlorophyll-deficient. Interestingly, approximately 45% of stomata had an unusual, previously not-described, morphology of thin-shaped chlorophyll-less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole-leaf photosynthetic parameters (PSII, qP, qN, FV '/FM' ) were comparable with wild-type plants. Time-resolved intact leaf gas-exchange analyses showed a reduction in stomatal conductance and CO2 -assimilation rates of the transgenic plants. Normalization of CO2 responses showed that stomata of transgenic plants respond to [CO2 ] shifts. Detailed stomatal aperture measurements of normal kidney-shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO2 ] elevation and abscisic acid (ABA), while thin-shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO2 ] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard cell CO2 and ABA signal transduction are not directly modulated by guard cell photosynthesis/electron transport. Moreover, the finding that chlorophyll-less stomata cause a 'deflated' thin-shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard cell turgor production. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Infection of cowpea protoplasts with sonchus yellow net virus and festuca leaf streak virus

NARCIS (Netherlands)

Beek, van N.A.M.

1986-01-01

The advantages of protoplast systems for plant virus research have been frequently reviewed (Zaitlin & Beachy, 1974; Takebe, 1975; Muhlbach, 1982; Sander & Mertens, 1984). Relatively little attention has been given to the limitations of such a system.

Protoplasts do not

La3+ uptake and its effect on the cytoskeleton in root protoplasts of Zea mays L.

Science.gov (United States)

Liu, Min; Hasenstein, Karl H

2005-03-01

La(3+) ions are known to antagonize Ca(2+) and are used as a Ca(2+) channel blocker but little is known on the direct effects of La(3+). Micromolar La(3+) concentrations promoted root growth while higher concentrations were inhibitory. The uptake of La(3+) in maize root protoplasts revealed a membrane binding component (0.14 and 0.44 pmol min(-1) protoplast(-1) for 100 and 1,000 microM La(3+)) followed by a slower concentration and time-dependent uptake. Uptake was reduced by Ca(2+), but had no substantial effect on other ions. La(3+) shifted microtubule organization from random to parallel but caused aggregation of microfilaments. Our data suggest that La(3+) is taken up into plant cells and affects growth via stabilization of the cytoskeleton.

Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion.

Science.gov (United States)

Patil, N S; Patil, S M; Govindwar, S P; Jadhav, J P

2015-02-01

Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste. The filamentous chitinolytic fungal strains A. oryzae NCIM 1272 and T. harzianum NCIM 1185 were selected as parents for protoplast fusion. Viable protoplasts were released from fungal mycelium using enzyme cocktail containing 5 mg ml(-1) lysing enzymes from T. harzianum, 0.06 mg ml(-1) β-glucuronidase from Helix pomatia and 1 mg ml(-1) purified Penicillium ochrochloron chitinase in 0.8 mol l(-1) sorbitol as an osmotic stabilizer. Intergeneric protoplast fusion was carried out using 60% polyethylene glycol as a fusogen. At optimum conditions, the regeneration frequency of the fused protoplasts on colloidal chitin medium and fusion frequency were calculated. Fusant showed higher rate of growth pattern, chitinase activity and protein content than parents. Fusant formation was confirmed by morphological markers, viz. colony morphology and spore size and denaturation gradient gel electrophoresis (DGGE). This study revealed protoplast fusion between A. oryzae and T. harzianum significantly enhanced chitinase activity which ultimately provides potential strain for degradation of shellfish waste. Consistency in the molecular characterization results using DGGE is the major outcome of this study which can be emerged as a fundamental step in fusant identification. Now it is need to provide attention over effective chitin degradation to manage shrimp processing issues. In this aspect, ability of fusant to degrade shellfish waste efficiently in short incubation time revealed discovery of potential strain in the reclamation of seafood processing crustacean bio-waste. © 2014 The Society for Applied Microbiology.

Phytochrome-mediated responses of cells and protoplasts of green calli obtained from the leaves of a CAM plant.

Science.gov (United States)

Mricha, A; Brulfert, J; Pierre, J N; Queiroz, O

1990-04-01

Green callus obtained from leaves of the CAM-inducible plant Kalanchoe blossfeldiana cv. Montezuma has previously been shown to perform C3-type photosynthesis under 16-h days and to shift to crassulacean acid metabolism (CAM) under 9-h days. The utilization of photoperiodic regimes (i.e. night interruptions by 30 min red light) established that CAM induction in the callus was under the control of phytochrome, as shown by measurements of CAM criteria: phosphoenolpyruvate carboxylase activity and malic acid pools. Short-term responsiveness of the callus cells to phytochrome modulations by monochromatic radiations was also established by the rapid changes observed in the diameter of the callus-derived protoplasts. These results provide further evidence that whole plant correlations are not necessary for phytochrome operativity.

Influence of protoplast fusion between two Trichoderma spp. on extracellular enzymes production and antagonistic activity.

Science.gov (United States)

Hassan, Mohamed M

2014-11-02

Biological control plays a crucial role in grapevine pathogens disease management. The cell-wall degrading enzymes chitinase, cellulase and β-glucanase have been suggested to be essential for the mycoparasitism activity of Trichoderma species against grapevine fungal pathogens. In order to develop a useful strain as a single source of these vital enzymes, it was intended to incorporate the characteristics of two parental fungicides tolerant mutants of Trichoderma belonging to the high chitinase producing species T. harzianum and the high cellulase producing species T. viride , by fusing their protoplasts. The phylogeny of the parental strains was carried out using a sequence of the 5.8S-ITS region. The BLAST of the obtained sequence identified these isolates as T. harzianum and T. viride . Protoplasts were isolated using lysing enzymes and were fused using polyethylene glycol. The fused protoplasts have been regenerated on protoplast regeneration minimal medium supplemented with two selective fungicides. Among the 40 fast growing fusants, 17 fusants were selected based on their enhanced growth on selective media for further studies. The fusant strains were growing 60%-70% faster than the parents up to third generation. All the 17 selected fusants exhibited morphological variations. Some fusant strains displayed threefold increased chitinase enzyme activity and twofold increase in β-glucanase enzyme activity compared to the parent strains. Most fusants showed powerful antagonistic activity against Macrophomin aphaseolina , Pythium ultimum and Sclerotium rolfsii pathogens. Fusant number 15 showed the highest inhibition percentage (92.8%) against M. phaseolina and P. ultimum, while fusant number 9 showed the highest inhibition percentage (98.2%) against the growth of S. rolfsii. A hyphal intertwining and degradation phenomenon was observed by scanning electron microscope. The Trichoderma antagonistic effect against pathogenic fungal mycelia was due to the

Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway?

Science.gov (United States)

Ma, Xiaohong; Shatil-Cohen, Arava; Ben-Dor, Shifra; Wigoda, Noa; Perera, Imara Y; Im, Yang Ju; Diminshtein, Sofia; Yu, Ling; Boss, Wendy F; Moshelion, Menachem; Moran, Nava

2015-03-01

Enhancing the membrane content of PtdInsP 2 , the already-recognized protein-regulating lipid, increased the osmotic water permeability of tobacco protoplasts, apparently by increasing the abundance of active aquaporins in their membranes. While phosphoinositides are implicated in cell volume changes and are known to regulate some ion channels, their modulation of aquaporins activity has not yet been reported for any organism. To examine this, we compared the osmotic water permeability (P f) of protoplasts isolated from tobacco (Nicotiana tabacum) cultured cells (NT1) with different (genetically lowered or elevated relative to controls) levels of inositol trisphosphate (InsP3) and phosphatidyl inositol [4,5] bisphosphate (PtdInsP2). To achieve this, the cells were transformed with, respectively, the human InsP3 5-phosphatase ('Ptase cells') or human phosphatidylinositol (4) phosphate 5-kinase ('PIPK cells'). The mean P f of the PIPK cells was several-fold higher relative to that of controls and Ptase cells. Three results favor aquaporins over the membrane matrix as underlying this excessive P f: (1) transient expression of the maize aquaporin ZmPIP2;4 in the PIPK cells increased P f by 12-30 μm s(-1), while in the controls only by 3-4 μm s(-1). (2) Cytosol acidification-known to inhibit aquaporins-lowered the P f in the PIPK cells down to control levels. (3) The transcript of at least one aquaporin was elevated in the PIPK cells. Together, the three results demonstrate the differences between the PIPK cells and their controls, and suggest a hitherto unobserved regulation of aquaporins by phosphoinositides, which could occur through direct interaction or indirect phosphoinositides-dependent cellular effects.

A comparison of different Gracilariopsis lemaneiformis (Rhodophyta) parts in biochemical characteristics, protoplast formation and regeneration

Science.gov (United States)

Wang, Zhongxia; Sui, Zhenghong; Hu, Yiyi; Zhang, Si; Pan, Yulong; Ju, Hongri

2014-08-01

Gracilariopsis lemaneiformis is a commercially exploited alga. Its filaceous thallus can be divided into three parts, holdfast, middle segment and tip. The growth and branch forming trend and agar content of these three parts were analyzed, respectively, in this study. The results showed that the tip had the highest growth rate and branched most, although it was the last part with branch forming ability. The holdfast formed branches earliest but slowly. Holdfast had the highest agar content. We also assessed the difference in protoplast formation and regeneration among three parts. The middle segment displayed the shortest enzymolysis time and the highest protoplast yield; whereas the tip had the strongest vitality of protoplasts formation. Juvenile plants were only obtained from the protoplasts generated from the tip. These results suggested that the differentiation and function of G. lemaneiformis was different.

Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill.) protoplast yield

OpenAIRE

Elżbieta Kuźniak; Marzena Wielanek; Urszula Małolepsza; Henryk Urbaniak

2013-01-01

The effects of soil or in vitro grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 107/g FW was obtained from leaves of in vitro grown plants. Low light intensity during donor plants in vitro culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to t...

Site-directed mutagenesis in Petunia × hybrida protoplast system using direct delivery of purified recombinant Cas9 ribonucleoproteins.

Science.gov (United States)

Subburaj, Saminathan; Chung, Sung Jin; Lee, Choongil; Ryu, Seuk-Min; Kim, Duk Hyoung; Kim, Jin-Soo; Bae, Sangsu; Lee, Geung-Joo

2016-07-01

Site-directed mutagenesis of nitrate reductase genes using direct delivery of purified Cas9 protein preassembled with guide RNA produces mutations efficiently in Petunia × hybrida protoplast system. The clustered, regularly interspaced, short palindromic repeat (CRISPR)-CRISPR associated endonuclease 9 (CRISPR/Cas9) system has been recently announced as a powerful molecular breeding tool for site-directed mutagenesis in higher plants. Here, we report a site-directed mutagenesis method targeting Petunia nitrate reductase (NR) gene locus. This method could create mutations efficiently using direct delivery of purified Cas9 protein and single guide RNA (sgRNA) into protoplast cells. After transient introduction of RNA-guided endonuclease (RGEN) ribonucleoproteins (RNPs) with different sgRNAs targeting NR genes, mutagenesis at the targeted loci was detected by T7E1 assay and confirmed by targeted deep sequencing. T7E1 assay showed that RGEN RNPs induced site-specific mutations at frequencies ranging from 2.4 to 21 % at four different sites (NR1, 2, 4 and 6) in the PhNR gene locus with average mutation efficiency of 14.9 ± 2.2 %. Targeted deep DNA sequencing revealed mutation rates of 5.3-17.8 % with average mutation rate of 11.5 ± 2 % at the same NR gene target sites in DNA fragments of analyzed protoplast transfectants. Further analysis from targeted deep sequencing showed that the average ratio of deletion to insertion produced collectively by the four NR-RGEN target sites (NR1, 2, 4, and 6) was about 63:37. Our results demonstrated that direct delivery of RGEN RNPs into protoplast cells of Petunia can be exploited as an efficient tool for site-directed mutagenesis of genes or genome editing in plant systems.

Guard Cell and Tropomyosin Inspired Chemical Sensor

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Jacquelyn K.S. Nagel

2013-10-01

Full Text Available Sensors are an integral part of many engineered products and systems. Biological inspiration has the potential to improve current sensor designs as well as inspire innovative ones. This paper presents the design of an innovative, biologically-inspired chemical sensor that performs “up-front” processing through mechanical means. Inspiration from the physiology (function of the guard cell coupled with the morphology (form and physiology of tropomyosin resulted in two concept variants for the chemical sensor. Applications of the sensor design include environmental monitoring of harmful gases, and a non-invasive approach to detect illnesses including diabetes, liver disease, and cancer on the breath.

Liposome-enhanced transformation of Streptococcus lactis and plasmid transfer by intergeneric protoplast fusion of Streptococcus lactis and Bacillus subtilis

NARCIS (Netherlands)

Vossen, Jos M.B.M. van der; Kok, Jan; Lelie, Daniel van der; Venema, Gerhardus

An efficient protoplast transformation system and a procedure of plasmid transfer by means of protoplast fusion is described for Streptococcus lactis. Protoplasts of S. lactis IL1403 and S. lactis MG1363 were transformed by pGK12 [2.9 MDa erythromycin resistance (Emr)] with an efficiency of 3 × 10^5

Analysis of Microbe-Associated Molecular Pattern-Responsive Synthetic Promoters with the Parsley Protoplast System.

Science.gov (United States)

Kanofsky, Konstantin; Lehmeyer, Mona; Schulze, Jutta; Hehl, Reinhard

2016-01-01

Plants recognize pathogens by microbe-associated molecular patterns (MAMPs) and subsequently induce an immune response. The regulation of gene expression during the immune response depends largely on cis-sequences conserved in promoters of MAMP-responsive genes. These cis-sequences can be analyzed by constructing synthetic promoters linked to a reporter gene and by testing these constructs in transient expression systems. Here, the use of the parsley (Petroselinum crispum) protoplast system for analyzing MAMP-responsive synthetic promoters is described. The synthetic promoter consists of four copies of a potential MAMP-responsive cis-sequence cloned upstream of a minimal promoter and the uidA reporter gene. The reporter plasmid contains a second reporter gene, which is constitutively expressed and hence eliminates the requirement of a second plasmid used as a transformation control. The reporter plasmid is transformed into parsley protoplasts that are elicited by the MAMP Pep25. The MAMP responsiveness is validated by comparing the reporter gene activity from MAMP-treated and untreated cells and by normalizing reporter gene activity using the constitutively expressed reporter gene.

Metabolomic Responses of Guard Cells and Mesophyll Cells to Bicarbonate

Science.gov (United States)

Misra, Biswapriya B.; de Armas, Evaldo; Tong, Zhaohui; Chen, Sixue

2015-01-01

Anthropogenic CO2 presently at 400 ppm is expected to reach 550 ppm in 2050, an increment expected to affect plant growth and productivity. Paired stomatal guard cells (GCs) are the gate-way for water, CO2, and pathogen, while mesophyll cells (MCs) represent the bulk cell-type of green leaves mainly for photosynthesis. We used the two different cell types, i.e., GCs and MCs from canola (Brassica napus) to profile metabolomic changes upon increased CO2 through supplementation with bicarbonate (HCO3 -). Two metabolomics platforms enabled quantification of 268 metabolites in a time-course study to reveal short-term responses. The HCO3 - responsive metabolomes of the cell types differed in their responsiveness. The MCs demonstrated increased amino acids, phenylpropanoids, redox metabolites, auxins and cytokinins, all of which were decreased in GCs in response to HCO3 -. In addition, the GCs showed differential increases of primary C-metabolites, N-metabolites (e.g., purines and amino acids), and defense-responsive pathways (e.g., alkaloids, phenolics, and flavonoids) as compared to the MCs, indicating differential C/N homeostasis in the cell-types. The metabolomics results provide insights into plant responses and crop productivity under future climatic changes where elevated CO2 conditions are to take center-stage. PMID:26641455

ISOLATION OF MESOPHYLL PROTOPLASTS FROM MEDITERRANEAN WOODY PLANTS FOR THE STUDY OF DNA INTEGRITY UNDER ABIOTIC STRESS

Directory of Open Access Journals (Sweden)

Elena Kuzminsky

2016-08-01

Full Text Available Abiotic stresses have considerable negative impact on Mediterranean plant ecosystems and better comprehension of the genetic control of response and adaptation of trees to global changes is urgently needed. The Single Cell Gel Electrophoresis assay could be considered a good estimator of DNA damage in an individual eukaryotic cell. This method has been mainly employed in animal tissues, because the plant cell wall represents an obstacle for the extraction of nuclei; moreover, in Mediterranean woody species, especially in the sclerophyll plants, this procedure can be quite difficult because of the presence of sclerenchyma and hardened cells. On the other hand, these plants represent an interesting material to be studied because of the ability of these plants to tolerate abiotic stress. For instance, holm oak (Quercus ilex L. has been selected as the model plant to identify critical levels of O3 for Southern European forests. Consequently, a quantitative method for the evaluation of cell injury of leaf tissues of this species is required. Optimal conditions for high-yield nuclei isolation were obtained by using protoplast technology and a detailed description of the method is provided and discussed. White poplar (Populus alba L. was used as an internal control for protoplast isolation. Such a method has not been previously reported in newly fully developed leaves of holm oak. This method combined with Single Cell Gel Electrophoresis assay represents a new tool for testing the DNA integrity of leaf tissues in higher plants under stress conditions.

Comparative conventional- and quantum dot-labelling strategies for LPS binding site detection in Arabidopsis thaliana mesophyll protoplasts

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Londiwe Siphephise Mgcina

2015-05-01

Full Text Available Lipopolysaccharide (LPS from Gram-negative bacteria is recognized as a microbe-associated molecular pattern (MAMP and not only induces an innate immune response in plants, but also stimulates the development of characteristic defense responses. However, identification and characterization of a cell surface LPS-receptor/binding site, as described in mammals, remains elusive in plants. As an amphiphilic, macromolecular lipoglycan, intact LPS potentially contains three MAMP-active regions, represented by the O-polysaccharide chain, the core and the lipid A. Binding site studies with intact labelled LPS were conducted in Arabidopsis thaliana protoplasts and quantified using flow cytometry fluorescence changes. Qdots, which allow non-covalent, hydrophobic labelling were used as a novel strategy in this study and compared to covalent, hydrophilic labelling with Alexa 488. Affinity for LPS-binding sites was clearly demonstrated by concentration-, temperature- and time-dependent increases in protoplast fluorescence following treatment with the labelled LPS. Moreover, this induced fluorescence increase was convincingly reduced following pre-treatment with excess unlabeled LPS, thereby indicating reversibility of LPS binding. Inhibition of the binding process is also reported using endo- and exocytosis inhibitors. Here, we present evidence for the anticipated presence of LPS-specific binding sites in Arabidopsis protoplasts, and furthermore propose Qdots as a more sensitive LPS-labelling strategy in comparison to the conventional Alexa 488 hydrazide label for binding studies.

Viral protein synthesis in cowpea mosaic virus infected protoplasts

International Nuclear Information System (INIS)

Rottier, P.

1980-01-01

Some aspects of cowpea mosaic virus (CPMV) multiplication in cowpea mesophyll protoplasts were studied. The detection and characterization of proteins whose synthesis is induced or is stimulated upon virus infection was performed with the aid of radioactive labelling. (Auth.)

Novel thermostable clostridial strains through protoplast fusion for enhanced biobutanol production at higher temperature—preliminary study

Directory of Open Access Journals (Sweden)

Muhammad Ferhan

2016-01-01

Full Text Available The objective of this study is to improve the thermal stability of clostridium strains for enhanced biobutanol production. Thermostable clostridia species were developed through protoplast fusion between mesophilic clostridial species (i.e., Clostridium beijerinckii and Clostridium acetobutylicum and thermophilic clostridial species (i.e., Clostridium thermocellum. Production of biobutanol was examined in the present preliminary study using the clostridium strains and their protoplast fusants using sugar mixture with composition identical to that of wheat straw acid hydrolysate. Maximum biobutanol production of 9.4 g/L was achieved by a fused strain at 45 °C with total sugar consumption of 66% compared to that at 35 °C (i.e., 8.4 g/L production and 64% total sugar consumption. Glucose and xylose uptake rates were generally higher compared to all other individual sugars in the feedstock. In general, average cell concentrations were in close proximity for all parenting and fused strains at 35 °C; i.e., in the range of 5.12 × 107 to 5.49 × 107 cells/mL. Average cell concentration of fusants between the mesophilic clostridial species and the thermophilic clostridial species slightly increased to ~ 5.62 × 107 cells/mL at a higher temperature of 45 °C. These results, in addition to the ones obtained for the butanol production, demonstrate enhanced thermal stability of both fusants at a higher temperature (45 °C.

Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax.

Science.gov (United States)

Pigna, Gaia; Dhillon, Taniya; Dlugosz, Elizabeth M; Yuan, Joshua S; Gorman, Connor; Morandini, Piero; Lenaghan, Scott C; Stewart, C Neal

2016-12-01

Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 µM 2,4-D and 4.4 µM BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 µM 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 ± 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Improvement of polysaccharide and triterpenoid production of Ganoderma lucidum through mutagenesis of protoplasts

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Ren Peng

2016-03-01

Full Text Available Ganoderma lucidum is a traditional medicinal macrofungus in China, which has two kinds of key bioactive compounds -- polysaccharides and triterpenoids. To improve the polysaccharide and triterpenoid production from G. lucidum, the preparation and regeneration conditions of protoplasts were optimized. This was done by systematic trials with various parameters, and protoplast mutation was subsequently performed. A mycelium that was cultivated for seven days and treated with 0.33 mL of 1% snailase and 0.66 mL of 0.5% cellulase solution for 2.5 h at 30 °C in the presence of osmotic pressure stabilizer mannitol (0.5 mol/L, had the best conditions, in which the resultant protoplasts were 6.40 × 105/mL and the regeneration rate was 6.25%. The resultant protoplasts were subjected to subsequent mutation by lithium chloride or by the combination of lithium chloride and Triton X-100. The highest yields of intracellular polysaccharide and triterpenoid in two mutant strains were 37.50 and 40.81 mg/g, which were increased with 568.45% and 373.43%, respectively, as compared to the original strain. Furthermore, the yields of intracellular polysaccharides and triterpenoids in the second generation and the third generation of the mutants were comparable to that of the first generation, which showed genetic stability of the mutants for the production of polysaccharides and triterpenoids.

A Protoplast Transient Expression System to Enable Molecular, Cellular, and Functional Studies in Phalaenopsis orchids

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Hsiang-Yin Lin

2018-06-01

Full Text Available The enigmatic nature of the specialized developmental programs of orchids has fascinated plant biologists for centuries. The recent releases of orchid genomes indicate that orchids possess new gene families and family expansions and contractions to regulate a diverse suite of developmental processes. However, the extremely long orchid life cycle and lack of molecular toolkit have hampered the advancement of orchid biology research. To overcome the technical difficulties and establish a platform for rapid gene regulation studies, in this study, we developed an efficient protoplast isolation and transient expression system for Phalaenopsis aphrodite. This protocol was successfully applied to protein subcellular localization and protein–protein interaction studies. Moreover, it was confirmed to be useful in delineating the PaE2F/PaDP-dependent cell cycle pathway and studying auxin response. In summary, the established orchid protoplast transient expression system provides a means to functionally characterize orchid genes at the molecular level allowing assessment of transcriptome responses to transgene expression and widening the scope of molecular studies in orchids.

Plant regeneration from protoplasts ofVicia narbonensis via somatic embryogenesis and shoot organogenesis.

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Tegeder, M; Kohn, H; Nibbe, M; Schieder, O; Pickardt, T

1996-11-01

Protoplasts ofVicia narbonensis isolated from epicotyls and shoot tips of etiolated seedlings were embedded in 1.4% sodium-alginate at a final density of 2.5×10(5) protoplasts/ml and cultivated in Kao and Michayluk-medium containing 0.5 mg/I of each of 2,4- dichlorophenoxyacetic acid, naphthylacetic acid and 6 -benzylaminopurine. A division frequency of 36% and a plating efficiency of 0.40-0.5% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred onto gelrite-solidified Murashige and Skoog-media containing various growth regulators. Regeneration of plants was achieved via two morphologically distinguishable pathways. A two step protocol (initially on medium with a high auxin concentration followed by a culture phase with lowered auxin amount) was used to regenerate somatic embryos, whereas cultivation on medium containing thidiazuron and naphthylacetic acid resulted in shoot morphogenesis. Mature plants were recovered from both somatic embryos as well as from thidiazuron-induced shoots.

Effect of Radiation Dosage on Efficiency of Chloroplast Transfer by Protoplast Fusion in Nicotiana

OpenAIRE

Menczel, László; Galiba, Gábor; Nagy, Ferenc; Maliga, Pál

1982-01-01

Chloroplasts of Nicotiana tabacum SR1 were transferred into Nicotiana plumbaginifolia by protoplast fusion. The protoplasts of the organelle donor were irradiated with different lethal doses using a 60Co source, to facilitate the elimination of their nuclei from the fusion products. After fusion induction, clones derived from fusion products and containing streptomycin-resistant N. tabacum SR1 chloroplasts were selected by their ability to green on a selective medium. When N. tabacum protopla...

Hydrolytic enzymes in the central vacuole of plant cells

International Nuclear Information System (INIS)

Boller, T.; Kende, H.

1979-01-01

The hydrolase content of vacuoles isolated from protoplasts of suspension-cultured tobacco cells, of tulip petals, and pineapple leaves, and the sedimentation behavior of tobacco tonoplasts were studied. Three precautions were found to be important for the analysis of vacuolar hydrolases and of the tonoplast: (a) purification of protoplasts in a Ficoll gradient was necessary to remove cell debris which contained contaminating hydrolases adsorbed from the fungal cell-wall-degrading enzyme preparation; (b) hydrolase activities in the homogenates of the intact cells or the tissue used and of the purified protoplasts had to be compared to verify the absence of contaminating hydrolases in the protoplast preparation; and (c) vacuoles obtained from the protoplasts by an osmotic shock had to be purified from the lysate in a Ficoll gradient. Since the density of the central vacuole approximates that of the protoplasts, about a 10% contamination of the vacuolar preparation by surviving protoplasts could not be eliminated. The intracellular activities of the following acid hydrolases were primarily localized in the vacuole of tobacco cells: α-mannosidase, β-N-acetylglucosaminidase, β-fructosidase, nuclease, phosphatase, phosphodiesterase. A similar composition of acid hydrolases was found in vacuoles obtained from protoplasts of tulip petals. Proteinase, a hydrolase with low activity in tobacco cells and tulip petals was found to be vacuolar in pineapple leaves, a tissue containing high levels of this enzyme. None of the vacuolar enzymes investigated ws found to be bound to the tonoplast. When vacuoles were isolated from cells labeled with radioactive choline, the vacuolar membrane was found to contain radioactivity. On sucrose gradients, the label incorporated into tonoplasts banded around a density of 1.10 grams per cubic centimeter

The barley anion channel, HvALMT1, has multiple roles in guard cell physiology and grain metabolism.

Science.gov (United States)

Xu, Muyun; Gruber, Benjamin D; Delhaize, Emmanuel; White, Rosemary G; James, Richard A; You, Jiangfeng; Yang, Zhenming; Ryan, Peter R

2015-01-01

The barley (Hordeum vulgare) gene HvALMT1 encodes an anion channel in guard cells and in certain root tissues indicating that it may perform multiple roles. The protein localizes to the plasma membrane and facilitates malate efflux from cells when constitutively expressed in barley plants and Xenopus oocytes. This study investigated the function of HvALMT1 further by identifying its tissue-specific expression and by generating and characterizing RNAi lines with reduced HvALMT1 expression. We show that transgenic plants with 18-30% of wild-type HvALMT1 expression had impaired guard cell function. They maintained higher stomatal conductance in low light intensity and lost water more rapidly from excised leaves than the null segregant control plants. Tissue-specific expression of HvALMT1 was investigated in developing grain and during germination using transgenic barley lines expressing the green fluorescent protein (GFP) with the HvALMT1 promoter. We found that HvALMT1 is expressed in the nucellar projection, the aleurone layer and the scutellum of developing barley grain. Malate release measured from isolated aleurone layers prepared from imbibed grain was significantly lower in the RNAi barley plants compared with control plants. These data provide molecular and physiological evidence that HvALMT1 functions in guard cells, in grain development and during germination. We propose that HvALMT1 releases malate and perhaps other anions from guard cells to promote stomatal closure. The likely roles of HvALMT1 during seed development and grain germination are also discussed. © 2014 Scandinavian Plant Physiology Society.

The Arabidopsis guard cell outward potassium channel GORK is regulated by CPK33.

Science.gov (United States)

Corratgé-Faillie, Claire; Ronzier, Elsa; Sanchez, Frédéric; Prado, Karine; Kim, Jeong-Hyeon; Lanciano, Sophie; Leonhardt, Nathalie; Lacombe, Benoît; Xiong, Tou Cheu

2017-07-01

A complex signaling network involving voltage-gated potassium channels from the Shaker family contributes to the regulation of stomatal aperture. Several kinases and phosphatases have been shown to be crucial for ABA-dependent regulation of the ion transporters. To date, the Ca 2+ -dependent regulation of Shaker channels by Ca 2+ -dependent protein kinases (CPKs) is still elusive. A functional screen in Xenopus oocytes was launched to identify such CPKs able to regulate the three main guard cell Shaker channels KAT1, KAT2, and GORK. Seven guard cell CPKs were tested and multiple CPK/Shaker couples were identified. Further work on CPK33 indicates that GORK activity is enhanced by CPK33 and unaffected by a nonfunctional CPK33 (CPK33-K102M). Furthermore, Ca 2+ -induced stomatal closure is impaired in two cpk33 mutant plants. © 2017 Federation of European Biochemical Societies.

Factors affecting callus and protoplast production and regeneration of plants from garlic tissue cultures

International Nuclear Information System (INIS)

Al-Safadi, B.; Nabulsi, I.

2001-08-01

Five cultivars of garlic, two explants, six callusing media, six regeneration media, two kinds of light and several doses of gamma irradiation were used to determine the best conditions for callus induction and plant regeneration from garlic tissue cultures. Also, some experiments were conducted to study the possibility to isolate protoplast and regenerate plants. The experiment showed that medium MS9 was good for regenerating plant directly from basal plate without going through callus phase. ANOVA exhibited significant differences among used cultivars in their ability to form callus. No significant difference was observed between 16 hr light and complete darkness in callus growth. However, appearance of callus was generally better on darkness. Cultivar varied in their ability to regenerate and interaction between cultivars and media was observed. Cultivar kisswany was the best in regeneration (38%) and medium MS47 was the best among used media (35%). Light type played a significant role in regeneration of plants where red light was much better than white light in inducing regeneration (68% vs 36%). ANOVA revealed significant effect of low doses of gamma irradiation on stimulation regeneration of plant whereas high doses prevented regeneration. Many experiments were conducted to isolate protoplast and regenerate plants. The best method for culturing was the droplet and the best conditions for incubation were complete darkness at 25 Degreed centigrade. This lead to formation of cell wall but no cell division was observed (author)

Conditions de sélection in vitro de cals issus des disques foliaires et des protoplastes de Pelargonium tolérant plus la sécheresse

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M. MOKHTARI

2015-01-01

Full Text Available To induce drought resistance, callus from leaf discs and protoplast of Pelargonium x hortorum (‘Deep Salmon’ and ‘Panaché Sud’ were grown in vitro in osmoticum rich medium. Percent survival of the callus varied with growth medium, variety and concentration of osmoticum. Compared to protoplasts, leaf discs were simple to handle. However, protoplasts growing requires enzymatic additives and delicate procedures. The protoplasts viability was 86% for ‘Deep Salmon’ and 90% for ‘Panaché Sud’. The yield was 5.67 x 106 protoplasts / g FM for ‘Deep Salmon’ and 11.35 x 106 for ‘Panaché Sud’. The callus from leaf discs of the variety Deep Salmon survived a maximum concentration of 0.5 M sucrose and 0.27 M mannitol or sorbitol. A dose of 0.6 M sucrose was the threshold limit for the survival of 12.5% ​​protoplasts with a division ratio of 2% for ‘Deep Salmon’ and 18.7% of protoplasts with a division ratio of 3.2% for ‘Panaché Sud’. For the mannitol, the maximum limit was 0.6 M for a 13.5% viability of protoplasts with a division ratio 3.6% for ‘Deep Salmon’ and 16.1% of protoplasts with a division factor 3 % respectively for ‘Panaché Sud’. The 20% PEG allowed the survival of 21.1% protoplast and a division rate of 0.2% in ‘Deep Salmon’, but it has totally inhibited protoplast division of ‘Panaché Sud’, even at 5%.

Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

NARCIS (Netherlands)

Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P.; de, Vicente A.

A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool

Response of haploid and diploid protoplasts from Datura innoxia Mill. and Petunia hybrida L. to treatment with X-rays and a chemical mutagen

International Nuclear Information System (INIS)

Krumbiegel, G.

1979-01-01

Haploid and diploid protoplasts of the two Solanaceous species Datura innoxia Mill. and Petunia Hybridia L., were exposed to two different mutagens, increased doses of X-rays and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). With both species the survival rates of haploid protoplasts decreased exponentially with increased doses of X-rays and increased concentrations of MNNG. Diploid protoplasts showed a higher resistance than haploids only at higher mutagen doses or concentrations. After the MNNG-treatment of haploid protoplasts from Datura innoxia, four mutants with altered pigment patterns were isolated. (author)

Hydrolytic enzymes in the central vacuole of plant cells.

Science.gov (United States)

Boller, T; Kende, H

1979-06-01

The hydrolase content of vacuoles isolated from protoplasts of suspension-cultured tobacco cells, of tulip petals, and of pineapple leaves, and the sedimentation behavior of tobacco tonoplasts were studied. Three precautions were found to be important for the analysis of vacuolar hydrolases and of the tonoplast. (a) Purification of protoplasts in a Ficoll gradient was necessary to remove cell debris which contained contaminating hydrolases adsorbed from the fungal cell-wall-degrading enzyme preparation. (b) Hydrolase activities in the homogenates of the intact cells or the tissue used and of the purified protoplasts had to be compared to verify the absence of contaminating hydrolases in the protoplast preparation. (c) Vacuoles obtained from the protoplasts by an osmotic shock had to be purified from the lysate in a Ficoll gradient. Since the density of the central vacuole approximates that of the protoplasts, about a 10% contamination of the vacuolar preparation by surviving protoplasts could not be eliminated and had to be taken into account when the distribution of enzymes and of radioactivity was calculated.THE INTRACELLULAR ACTIVITIES OF THE FOLLOWING ACID HYDROLASES WERE PRIMARILY LOCALIZED IN THE VACUOLE OF TOBACCO CELLS: alpha-mannosidase, beta-N-acetylglucosaminidase, beta-fructosidase, nuclease, phosphatase, phosphodiesterase. A similar composition of acid hydrolases was found in vacuoles obtained from protoplasts of tulip petals. Proteinase, a hydrolase with low activity in tobacco cells and tulip petals and therefore difficult to localize unequivocally, was found to be vacuolar in pineapple leaves, a tissue containing high levels of this enzyme. Our data support the hypothesis that the central vacuole of higher plant cells has an enzyme composition analogous to that of the animal lysosome.None of the vacuolar enzymes investigated was found to be bound to the tonoplast. When vacuoles were isolated from cells labeled with radioactive choline, the vacuolar

High efficiency protoplast isolation from in vitro cultures and hairy ...

African Journals Online (AJOL)

In vitro cultures of the medicinal plant Maesa lanceolata were established to enable the cultivation of plant material for the production of protoplasts. Callus cultures were initiated using leaves collected from shoot cultures and the root tips from hairy root cultures obtained upon Agrobacterium rhizogenes transformation.

Overexpression of plasma membrane H+-ATPase in guard cells promotes light-induced stomatal opening and enhances plant growth.

Science.gov (United States)

Wang, Yin; Noguchi, Ko; Ono, Natsuko; Inoue, Shin-ichiro; Terashima, Ichiro; Kinoshita, Toshinori

2014-01-07

Stomatal pores surrounded by a pair of guard cells in the plant epidermis control gas exchange between plants and the atmosphere in response to light, CO2, and the plant hormone abscisic acid. Light-induced stomatal opening is mediated by at least three key components: the blue light receptor phototropin (phot1 and phot2), plasma membrane H(+)-ATPase, and plasma membrane inward-rectifying K(+) channels. Very few attempts have been made to enhance stomatal opening with the goal of increasing photosynthesis and plant growth, even though stomatal resistance is thought to be the major limiting factor for CO2 uptake by plants. Here, we show that transgenic Arabidopsis plants overexpressing H(+)-ATPase using the strong guard cell promoter GC1 showed enhanced light-induced stomatal opening, photosynthesis, and plant growth. The transgenic plants produced larger and increased numbers of rosette leaves, with ∼42-63% greater fresh and dry weights than the wild type in the first 25 d of growth. The dry weights of total flowering stems of 45-d-old transgenic plants, including seeds, siliques, and flowers, were ∼36-41% greater than those of the wild type. In addition, stomata in the transgenic plants closed normally in response to darkness and abscisic acid. In contrast, the overexpression of phototropin or inward-rectifying K(+) channels in guard cells had no effect on these phenotypes. These results demonstrate that stomatal aperture is a limiting factor in photosynthesis and plant growth, and that manipulation of stomatal opening by overexpressing H(+)-ATPase in guard cells is useful for the promotion of plant growth.

Basidiospore and Protoplast Regeneration from Raised Fruiting Bodies of Pathogenic Ganoderma boninense.

Science.gov (United States)

Govender, Nisha T; Mahmood, Maziah; Seman, Idris A; Mui-Yun, Wong

2016-08-26

Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological potentials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from Trichoderma harzianum (10 mg/ml) suspended in osmotic stabilizer (0.6 M potassium chloride and 20 mM dipotassium phosphate buffer) yielded the highest number of viable protoplasts (8.9 × 106 single colonies) among all possible combinations tested (regeneration media, age of mycelium, osmotic stabilizer, digestive enzyme and incubation period).

Zeaxanthin concentrations co-segregate with the magnitude of the blue light response of adaxial guard cells and leaf stomatal conductances in an F2 population of pima cotton

Energy Technology Data Exchange (ETDEWEB)

Quinones, M.A.; Lu Zhenmin; Zeiger, E. (Univ. of California, Los Angeles (United States))

1993-05-01

A blue light (BL) response of adaxial (AD) guard cells was investigated in two cotton lines with contrasting rates of stomatal conductances (g). This response is expressed as an enhancement of the red light-induced chlorophyll a fluorescence quenching by BL, and has an action spectrum indicative of a carotenoid photoreceptor. Ad guard cell from the high g, advanced line Pima S-6 have a higher carotenoid content and a larger BL response than those from the low g, primitive cotton, B368. In a growth chamber-grown F2 population of a cross between the two lines (n=30), g of individual plants segregated over a range exceeding the average g of the parental populations. Carotenoid content and the BL response of ad guard cell also segregated. There was a positive, strong correlation (r=0.71) between leaf g and the magnitude of the BL response of ad guard cells, indicating that both parameters are under genetic control, and that the BL response of guard cells contributes to the modulation of g. The concentration of all xanthopylls and [beta]-carotene in the ad guard cells correlated poorly with the BL response, except for zeaxanthin (r=0.71). In all green systems, xanthophylls are located inside the chloroplast which suggests that zeaxanthin functions in these organelle as a blue light photoreceptor for cotton guard cells.

Guard News - The National Guard

Science.gov (United States)

operations during chemical attack May 25, 2018 - SWANTON, Ohio - Maintenance is a critical component in Leaders discuss future of National Guard cyber warfare May 21, 2018 - CAMP ATTERBURY, Ind. - While National Guard teams from across the nation tested themselves on their cyber defensive capabilities at

Uniformity of plants regenerated from orange (Citrus sinensis Osb.) protoplasts.

Science.gov (United States)

Kobayashi, S

1987-05-01

Using 25 plants (protoclones) regenerated from orange (Citrus sinensis Osb.) protoplasts, several characters, including leaf and flower morphology, leaf oil, isozyme patterns and chromosome number, were examined. No significant variations in each character were recorded among the protoclones. Uniformity observed among protoclones was identical to that of nucellar seedlings.

National Guard > About the Guard > Today in Guard History

Science.gov (United States)

National Guard About Us By the Numbers Contact Us FAQ Federal Mission History Join Us Leaders Director of Program Training & Technology Battle Lab (T3BL) Civil Support Simulation Exercises Regional Training March Today in Guard History Leadership CNGB VCNGB SEA DANG DARNG Joint Staff J-1 J-2 J-3 J-4 J-5 J-6 J

Isolation and culture of protoplasts of Côte d'Ivoire's pearl millet ...

African Journals Online (AJOL)

SARAH

2015-08-31

. Journal of Biology and Chemical Science 8 (5):. 2222-2231. Timbo de Oliveira AL, Davide LC, Pereira Pinto JEB,. Pereira AV, 2010. Protoplast production from. Napier grass and Pearl millet triploid hybrids.Ciens.Agrotec.

Polyamine binding to proteins in oat and Petunia protoplasts

Science.gov (United States)

Mizrahi, Y.; Applewhite, P. B.; Galston, A. W.

1989-01-01

Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants. To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa). In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action. In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH. The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS. Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons. Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.

Naturally Induced Secretions of the Potato Cyst Nematode Co-stimulate the Proliferation of Both Tobacco Leaf Protoplasts and Human Peripheral Blood Mononuclear Cells

NARCIS (Netherlands)

Goverse, A.; Rouppe van der Voort, J.N.A.M.; Rouppe van der voort, C.; Kavelaars, A.; Smant, G.; Schots, A.; Bakker, J.; Helder, J.

1999-01-01

Naturally induced secretions from infective juveniles of the potato cyst nematode Globodera rostochiensis co-stimulate the proliferation of tobacco leaf protoplasts in the presence of the synthetic phytohormones α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). With the use of a

Radiosensitivity of Nicotiana protoplasts. Action on cell; cycle effects of low dose and fractionated irradiations; biological repair

International Nuclear Information System (INIS)

Magnien, E.

1981-10-01

Leaf protoplasts of Nicotiana plumbaginifolia and Nicotiana sylvestris demonstrate five main qualities: they can be maintained as haploid lines; they constitute starting populations with a remarkable cytological homogeneity; they show a transient initial lag-phase; they yield very high plating efficiencies and retain permanently a complete differentiation capacity; being derived of a cell wall, they appear well adapted for fusion experiments or enzymatic dosages. The resumption of mitotic activity was followed by cytophotometric measurements, labelling experiments, nuclear sizing and enzymatic assays. The action of 5 Gy gamma-ray irradiations delayed entrance in the S-phase, provoked an otherwise not verified dependency between transcription, translation and protein synthesis, increased nuclear volumes in the G2-phase, and slightly stimulated the activity of a repair enzyme. The plating efficiency was a sensitive end-point which allowed the evaluation of the biological effectiveness of low to medium radiation-doses after gamma-ray and fast neutron irradiations. The neutron dose-RBE relationship increased from 3 to 25 when the dose decreased from 5 Gy to 5 mGy. When fractionated into low single doses only, a neutron dose of 300 mGy markedly increased its biological effectiveness: this phenomenon could not be explained by cell progression, and necessitated additional hypotheses involving other mechanisms in the specific action of low radiation doses. Radiation-induced UDS was measured in presence of aphidicolin. A beta-like DNA-polymerase was shown to be definitely involved in nuclear repair synthesis [fr

Calcium specificity signaling mechanisms in abscisic acid signal transduction in Arabidopsis guard cells

Science.gov (United States)

Brandt, Benjamin; Munemasa, Shintaro; Wang, Cun; Nguyen, Desiree; Yong, Taiming; Yang, Paul G; Poretsky, Elly; Belknap, Thomas F; Waadt, Rainer; Alemán, Fernando; Schroeder, Julian I

2015-01-01

A central question is how specificity in cellular responses to the eukaryotic second messenger Ca2+ is achieved. Plant guard cells, that form stomatal pores for gas exchange, provide a powerful system for in depth investigation of Ca2+-signaling specificity in plants. In intact guard cells, abscisic acid (ABA) enhances (primes) the Ca2+-sensitivity of downstream signaling events that result in activation of S-type anion channels during stomatal closure, providing a specificity mechanism in Ca2+-signaling. However, the underlying genetic and biochemical mechanisms remain unknown. Here we show impairment of ABA signal transduction in stomata of calcium-dependent protein kinase quadruple mutant plants. Interestingly, protein phosphatase 2Cs prevent non-specific Ca2+-signaling. Moreover, we demonstrate an unexpected interdependence of the Ca2+-dependent and Ca2+-independent ABA-signaling branches and the in planta requirement of simultaneous phosphorylation at two key phosphorylation sites in SLAC1. We identify novel mechanisms ensuring specificity and robustness within stomatal Ca2+-signaling on a cellular, genetic, and biochemical level. DOI: http://dx.doi.org/10.7554/eLife.03599.001 PMID:26192964

Cell fusion as a tool for rice improvement

Energy Technology Data Exchange (ETDEWEB)

Hayashi, Y; Kyozuka, J; Terada, R; Nishibayashi, S; Shimamoto, K [Plantech Research Institute, Kamoshida, Midori-ku, Yokohama (Japan)

1990-01-01

Full text: Cell fusion offers a unique opportunity to hybridize sexually incompatible species and to mix cytoplasmic genomes in higher plants. Recent progress in plant regeneration from rice protoplasts facilitates an evaluation of the cell fusion method for rice improvement. By using electrofusion of protoplasts, we obtained hybrid/cybrid plants of the following combinations: Hybrids of rice x barnyard grass (E. oryzicola); Hybrids of rice x wild Oryza species; Cybrids of rice with transferred cms cytoplasm. For the latter, protoplasts irradiated with 70 krad x-rays were used. (author)

Isolamento e regeneração de protoplastos de Magnaporthe grisea Isolation and regeneration of Magnaporthe grisea protoplasts

Directory of Open Access Journals (Sweden)

Carlos Eduardo Marchi

2006-09-01

Full Text Available Protoplastos são ferramentas biológicas importantes para pesquisas em fungos filamentosos, sendo empregados intensamente em transformação genética. O isolamento de protoplastos de Magnaporthe grisea foi facilitado com Novozym 234, contudo, este complexo enzimático encontra-se indisponível no mercado. Assim, objetivou-se comparar a eficiência de enzimas líticas disponíveis comercialmente na obtenção de protoplastos de M. grisea. Paralelamente, analisaram-se estabilizadores osmóticos, tempos de digestão e freqüência de regeneração. Maior produção de protoplastos foi obtida com o uso simultâneo de Lysing Enzymes e Cellulase Onozuka R-10. O uso de 10 ou 15 mg de cada complexo enzimático, em 3 mL de estabilizador osmótico, resultou em maior liberação de protoplastos. O melhor estabilizador osmótico foi MgSO4 1,2 M / NaH2PO4 0,01 M, pH 5,8, seguido por MgSO4 0,8 M / NaH2PO4 0,01 M, pH 5,8. O isolamento de protoplastos foi monitorado a cada 60 minutos, atingindo o máximo após incubação por 3 a 6 horas. No entanto, maior freqüência de regeneração (19,4% foi registrada para protoplastos obtidos após 3 horas de hidrólise enzimática.Protoplasts are important biological tools in filamentous fungi research. Fungal protoplasts have been extensively used in experiments with genetic transformation. Protoplastization of Magnaporthe grisea was accomplished with Novozym 234, however, this enzymatic complex is no commercially available for purchase. Thus, the efficiency of several other commercial enzymes in M. grisea protoplasts preparation was investigated. At the same time, osmotic buffer, digestion time and regeneration rate were also analyzed. The highest protoplasts production was obtained with Lysing Enzymes plus Cellulase Onozuka R-10. The use of 10 or 15 mg of each enzymatic complex in 3 mL of osmotic buffer was most effective for the protoplasts yields. The best osmotic buffer was MgSO4 1.2 M / NaH2PO4 0.01 M, pH 5

Tomato protoplast DNA transformation : physical linkage and recombination of exogenous DNA sequences

NARCIS (Netherlands)

Jongsma, Maarten; Koornneef, Maarten; Zabel, Pim; Hille, Jacques

1987-01-01

Tomato protoplasts have been transformed with plasmid DNA's, containing a chimeric kanamycin resistance gene and putative tomato origins of replication. A calcium phosphate-DNA mediated transformation procedure was employed in combination with either polyethylene glycol or polyvinyl alcohol. There

Compound stress response in stomatal closure: a mathematical model of ABA and ethylene interaction in guard cells

Directory of Open Access Journals (Sweden)

Beguerisse-Dıaz Mariano

2012-11-01

Full Text Available Abstract Background Stomata are tiny pores in plant leaves that regulate gas and water exchange between the plant and its environment. Abscisic acid and ethylene are two well-known elicitors of stomatal closure when acting independently. However, when stomata are presented with a combination of both signals, they fail to close. Results Toshed light on this unexplained behaviour, we have collected time course measurements of stomatal aperture and hydrogen peroxide production in Arabidopsis thaliana guard cells treated with abscisic acid, ethylene, and a combination of both. Our experiments show that stomatal closure is linked to sustained high levels of hydrogen peroxide in guard cells. When treated with a combined dose of abscisic acid and ethylene, guard cells exhibit increased antioxidant activity that reduces hydrogen peroxide levels and precludes closure. We construct a simplified model of stomatal closure derived from known biochemical pathways that captures the experimentally observed behaviour. Conclusions Our experiments and modelling results suggest a distinct role for two antioxidant mechanisms during stomatal closure: a slower, delayed response activated by a single stimulus (abscisic acid ‘or’ ethylene and another more rapid ‘and’ mechanism that is only activated when both stimuli are present. Our model indicates that the presence of this rapid ‘and’ mechanism in the antioxidant response is key to explain the lack of closure under a combined stimulus.

Uncapped mRNA introduced into tobacco protoplasts can be imported into the nucleus and is trapped by leptomycin B.

Science.gov (United States)

Stuger, Rogier; Forreiter, Christoph

2004-08-01

The mechanism of nuclear export of RNAs in yeast and animal cells is rapidly being uncovered, but RNA export in plants has received little attention. We introduced capped and uncapped fluorescent mRNAs into tobacco (Nicotiana plumbaginifolia) protoplasts and studied their cellular localization. Following insertion, capped transcripts were found in the cytoplasm, while uncapped messengers transiently appeared in the nucleus in about one-quarter to one-third of the cells. These mRNAs were trapped by the nuclear export-inhibiting drug leptomycin B, pointing to an export mechanism in plants similar to Rev-NES-mediated RNP export in other organisms.

Expression analysis of Arabidopsis vacuolar sorting receptor 3 reveals a putative function in guard cells.

Science.gov (United States)

Avila, Emily L; Brown, Michelle; Pan, Songqin; Desikan, Radhika; Neill, Steven J; Girke, Thomas; Surpin, Marci; Raikhel, Natasha V

2008-01-01

Vacuolar sorting receptors (VSRs) are responsible for the proper targeting of soluble cargo proteins to their destination compartments. The Arabidopsis genome encodes seven VSRs. In this work, the spatio-temporal expression of one of the members of this gene family, AtVSR3, was determined by RT-PCR and promoter::reporter gene fusions. AtVSR3 was expressed specifically in guard cells. Consequently, a reverse genetics approach was taken to determine the function of AtVSR3 by using RNA interference (RNAi) technology. Plants expressing little or no AtVSR3 transcript had a compressed life cycle, bolting approximately 1 week earlier and senescing up to 2 weeks earlier than the wild-type parent line. While the development and distribution of stomata in AtVSR3 RNAi plants appeared normal, stomatal function was altered. The guard cells of mutant plants did not close in response to abscisic acid treatment, and the mean leaf temperatures of the RNAi plants were on average 0.8 degrees C lower than both wild type and another vacuolar sorting receptor mutant, atvsr1-1. Furthermore, the loss of AtVSR3 protein caused the accumulation of nitric oxide and hydrogen peroxide, signalling molecules implicated in the regulation of stomatal opening and closing. Finally, proteomics and western blot analyses of cellular proteins isolated from wild-type and AtVSR3 RNAi leaves showed that phospholipase Dgamma, which may play a role in abscisic acid signalling, accumulated to higher levels in AtVSR3 RNAi guard cells. Thus, AtVSR3 may play an important role in responses to plant stress.

Effects of ultraviolet radiation on viability of isolated Beta vulgaris and Hordeum vulgare protoplasts

International Nuclear Information System (INIS)

Bornman, J.F.; Bjoern, L.O.; Bornman, C.H.

1982-01-01

Estimates of viability as measured by vital straining with fluorescein diacetate were carried out on freshly isolated and partially aged (16-hour-old) Beta vulgaris and Hordeum vulgare mesophyll protoplasts following irradiation with UV-B. Damage to the photosynthetic system by UV-B was determined by delayed light emission (DLE). In the case of freshly isolated Protoplasts Beta was approximately 30% more susceptible than Hordeum following 3h irradiation, with viability decreasing from 90% to 40%. After storage of protoplasts on ice for 16 h UV-B radiation markedly depressed viability in both species, but in the case of Hordeum there was a substantial initial loss of nearly 70% in viability over the first hour of irradiation. The first 10 min of UV-B radiation decreased the intensity of DLE by 40% without appreciably affecting the decay rate. Longer treatment times did not give a proportional effect so that even after 60 min of UV-B the inhibition did not exceed 60%. This suggested that although the enzyme system responsible for FDA hydrolysis may be partially inactivated (viability was 75-80% as compared with 90% in the control), the UV-B did not penetrate the innermost parts of the chloroplasts, but left some thylakoids undamaged. (orig.)

Organization of cytoskeleton controls the changes in cytosolic calcium of cold-shocked Nicotiana plumbaginifolia protoplasts.

Science.gov (United States)

Mazars, C; Thion, L; Thuleau, P; Graziana, A; Knight, M R; Moreau, M; Ranjeva, R

1997-11-01

Using Nicotiana plumbaginifolia constitutively expressing the recombinant bioluminescent calcium indicator, aequorin, it has been previously demonstrated that plant cells react to cold-shock by an immediate rise in cytosolic calcium. Such an opportune system has been exploited to address the regulatory pathway involved in the calcium response. For this purpose, we have used protoplasts derived from N. plumbaginifolia leaves that behave as the whole plant but with a better reproducibility. By both immunodetecting cytoskeletal components on membrane ghosts and measuring the relative change in cytosolic calcium, we demonstrate that the organization of the cytoskeleton has profound influences on the calcium response. The disruption of the microtubule meshwork by various active drugs, such as colchicin, oryzalin and vinblastin, leads to an important increase in the cytosolic calcium (up to 400 nM) in cold-shocked protoplasts over control. beta-Lumicolchicin, an inactive analogue of colchicin, is ineffective either on cytoplasmic calcium increase or on microtubule organization. A microfilament disrupting drug, cytochalasin D, exerts a slight stimulatory effect, whereas the simultaneous disruption of microtubule and microfilament meshworks results in a dramatic increase in the calcium response to cold-shock. The results described in the present paper illustrate the role of the intracellular organization and, more specifically, the role of cytoskeleton in controlling the intensity of calcium response to an extracellular stimulus.

Plant adaptation to fluctuating environment and biomass production are strongly dependent on guard cell potassium channels

Science.gov (United States)

Lebaudy, Anne; Vavasseur, Alain; Hosy, Eric; Dreyer, Ingo; Leonhardt, Nathalie; Thibaud, Jean-Baptiste; Véry, Anne-Aliénor; Simonneau, Thierry; Sentenac, Hervé

2008-01-01

At least four genes encoding plasma membrane inward K+ channels (Kin channels) are expressed in Arabidopsis guard cells. A double mutant plant was engineered by disruption of a major Kin channel gene and expression of a dominant negative channel construct. Using the patch-clamp technique revealed that this mutant was totally deprived of guard cell Kin channel (GCKin) activity, providing a model to investigate the roles of this activity in the plant. GCKin activity was found to be an essential effector of stomatal opening triggered by membrane hyperpolarization and thereby of blue light-induced stomatal opening at dawn. It improved stomatal reactivity to external or internal signals (light, CO2 availability, and evaporative demand). It protected stomatal function against detrimental effects of Na+ when plants were grown in the presence of physiological concentrations of this cation, probably by enabling guard cells to selectively and rapidly take up K+ instead of Na+ during stomatal opening, thereby preventing deleterious effects of Na+ on stomatal closure. It was also shown to be a key component of the mechanisms that underlie the circadian rhythm of stomatal opening, which is known to gate stomatal responses to extracellular and intracellular signals. Finally, in a meteorological scenario with higher light intensity during the first hours of the photophase, GCKin activity was found to allow a strong increase (35%) in plant biomass production. Thus, a large diversity of approaches indicates that GCKin activity plays pleiotropic roles that crucially contribute to plant adaptation to fluctuating and stressing natural environments. PMID:18367672

Guarded Type Promotion

DEFF Research Database (Denmark)

Winther, Johnni

2011-01-01

conditional using the instanceof operator and thus the cast type is redundantly mentioned twice. We propose a new typing rule for Java called Guarded Type Promotion aimed at eliminating the need for the explicit casts when guarded. This new typing rule is backward compatible and has been fully implemented...... in a Java 6 compiler. Through our extensive testing of real-life code we show that guarded casts account for approximately one fourth of all casts and that Guarded Type Promotion can eliminate the need for 95 percent of these guarded casts....

Intracellular compartimentation of abscisic acid (ABA) in guard cells and mesophyll cells under exposure to SO sub 2. Kompartimentierung von Abscisinsaeure (ABA) in Schliess- und Mesophyllzellen unter SO sub 2 -Belastung

Energy Technology Data Exchange (ETDEWEB)

Baier, M.; Daeter, W.; Hartung, W. (Wuerzburg Univ. (Germany, F.R.). Lehrstuhl fuer Botanik 1)

1989-07-01

The effect of SO{sub 2} on the intracellular compartimentation of ABA in guard cells and mesophyll cells of Valerianella locusta was investigated, using the efflux compartmental analysis, as described by Behl and Hartung (1986). The cytoplasmic ABA content of the guard cells was reduced drastically by 6 {mu}molxm{sup -3} SO{sub 2} (20% of the controls). The vacuolar content was decreased less dramatically (70% of the controls). The ABA distribution of mesophyll cells remained uneffected by 6 {mu}molxm{sup -3} SO{sub 2}. The SO{sub 2} effects are explained by an acidification of the compartments. (orig.).

CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca(2+-permeable channels and stomatal closure.

Directory of Open Access Journals (Sweden)

Izumi C Mori

2006-10-01

Full Text Available Abscisic acid (ABA signal transduction has been proposed to utilize cytosolic Ca(2+ in guard cell ion channel regulation. However, genetic mutants in Ca(2+ sensors that impair guard cell or plant ion channel signaling responses have not been identified, and whether Ca(2+-independent ABA signaling mechanisms suffice for a full response remains unclear. Calcium-dependent protein kinases (CDPKs have been proposed to contribute to central signal transduction responses in plants. However, no Arabidopsis CDPK gene disruption mutant phenotype has been reported to date, likely due to overlapping redundancies in CDPKs. Two Arabidopsis guard cell-expressed CDPK genes, CPK3 and CPK6, showed gene disruption phenotypes. ABA and Ca(2+ activation of slow-type anion channels and, interestingly, ABA activation of plasma membrane Ca(2+-permeable channels were impaired in independent alleles of single and double cpk3cpk6 mutant guard cells. Furthermore, ABA- and Ca(2+-induced stomatal closing were partially impaired in these cpk3cpk6 mutant alleles. However, rapid-type anion channel current activity was not affected, consistent with the partial stomatal closing response in double mutants via a proposed branched signaling network. Imposed Ca(2+ oscillation experiments revealed that Ca(2+-reactive stomatal closure was reduced in CDPK double mutant plants. However, long-lasting Ca(2+-programmed stomatal closure was not impaired, providing genetic evidence for a functional separation of these two modes of Ca(2+-induced stomatal closing. Our findings show important functions of the CPK6 and CPK3 CDPKs in guard cell ion channel regulation and provide genetic evidence for calcium sensors that transduce stomatal ABA signaling.

A new mechanism for the regulation of stomatal aperture size in intact leaves: accumulation of mesophyll-derived sucrose in the guard-cell wall of Vicia faba

International Nuclear Information System (INIS)

Lu, P.; Outlaw, W.H. Jr.; Smith, B.G.; Freed, G.A.

1997-01-01

At various times after pulse-labeling broad bean (Vicia faba L.) leaflets with 14CO2, whole-leaf pieces and rinsed epidermal peels were harvested and subsequently processed for histochemical analysis. Cells dissected from whole leaf retained apoplastic contents, whereas those from rinsed peels contained only symplastic contents. Sucrose (Suc)-specific radioactivity peaked (111 GBq mol-1) in palisade cells at 20 min. In contrast, the 14C content and Suc-specific radioactivity were very low in guard cells for 20 min, implying little CO2 incorporation; both then peaked at 40 min. The guard-cell apoplast had a high maximum Suc-specific radioactivity (204 GBq mol-1) and a high Suc influx rate (0.05 pmol stoma-1 min-1). These and other comparisons implied the presence of (a) multiple Suc pools in mesophyll cells, (b) a localized mesophyll-apoplast region that exchanges with phloem and stomata, and (c) mesophyll-derived Suc in guard-cell walls sufficient to diminish stomatal opening by approximately 3 micrometers. Factors expected to enhance Suc accumulation in guard-cell walls are (a) high transpiration rate, which closes stomata, and (b) high apoplastic Suc concentration, which is elevated when mesophyll Suc efflux exceeds translocation. Therefore, multiple physiological factors are integrated in the attenuation of stomatal aperture size by this previously unrecognized mechanism

Effects of actonomycin D and ultraviolet irradiation on multiplication of brome mosaic virus in host and non-host cells

International Nuclear Information System (INIS)

Maekawa, K.; Furusawa, I.; Okuno, T.

1981-01-01

The modes of multiplication of brome mosaic virus (BMV) were compared in protoplasts isolated from host and non-host plants. BMV actively multiplied in the leaves and isolated mesophyll protoplasts of barley, a host of BMV. BMV multiplication in barley protoplasts was inhibited by addition of actinomycin D immediately after inoculation or by u.v. irradiation of the protoplasts before inoculation. In contrast, although BMV could not multiply in leaves of radish and turnip (non-hosts for BMV) it multiplied at a low level in protoplasts isolated from these two plant species. Moreover, u.v. irradiation, or the addition of actinomycin D, enhanced multiplication of BMV in radish and turnip protoplasts. These results suggest that (i) in the host cells replication of BMV is dependent on cellular metabolism of nucleic acid and protein, and (ii) in the non-host cells a substance(s) inhibitory to replication of BMV is synthesized. (author)

AGGREGATION AND FUSION OF PLANT-PROTOPLASTS AFTER SURFACE-LABELING WITH BIOTIN AND AVIDIN

NARCIS (Netherlands)

VANKESTEREN, WJP; MOLEMA, E; TEMPELAAR, MJ

1993-01-01

In mass electrofusion systems with aggregation of protoplasts by alignment, the yield and composition of fusion products can be predicted by a simple model. Through computer simulation, upper limits were found for the yield of binary and multi fusions. To overcome constraints on binary products,

Immuno and affinity cytochemical analysis of cell wall composition in the moss Physcomitrella patens

Directory of Open Access Journals (Sweden)

Elizabeth A. Berry

2016-03-01

Full Text Available In contrast to homeohydric vascular plants, mosses employ a poikilohydric strategy for surviving in the dry aerial environment. A detailed understanding of the structure, composition, and development of moss cell walls can contribute to our understanding of not only the evolution of overall cell wall complexity, but also the differences that have evolved in response to selection for different survival strategies. The model moss species Physcomitrella patens has a predominantly haploid lifecycle consisting of protonemal filaments that regenerate from protoplasts and enlarge by tip growth, and leafy gametophores composed of cells that enlarge by diffuse growth and differentiate into several different types. Advantages for genetic studies include methods for efficient targeted gene modification and extensive genomic resources. Immuno and affinity cytochemical labeling were used to examine the distribution of polysaccharides and proteins in regenerated protoplasts, protonemal filaments, rhizoids, and sectioned gametophores of P. patens. The cell wall composition of regenerated protoplasts was also characterized by flow cytometry. Crystalline cellulose was abundant in the cell walls of regenerating protoplasts and protonemal cells that developed on media of high osmolarity, whereas homogalacturonan was detected in the walls of protonemal cells that developed on low osmolarity media and not in regenerating protoplasts. Mannan was the major hemicellulose detected in all tissues tested. Arabinogalactan proteins were detected in different cell types by different probes, consistent with structural heterogeneity. The results reveal developmental and cell type specific differences in cell wall composition and provide a basis for analyzing cell wall phenotypes in knockout mutants.

Improved inhibitor tolerance in xylose-fermenting yeast Spathaspora passalidarum by mutagenesis and protoplast fusion

DEFF Research Database (Denmark)

Hou, Xiaoru; Yao, Shuo

2012-01-01

The xylose-fermenting yeast Spathaspora passalidarum showed excellent fermentation performance utilizing glucose and xylose under anaerobic conditions. But this yeast is highly sensitive to the inhibitors such as furfural present in the pretreated lignocellulosic biomass. In order to improve...... from fusion of the protoplasts of S. passalidarum M7 and a robust yeast, Saccharomyces cerevisiae ATCC 96581, were able to grow in 75% WSLQ and produce around 0.4 g ethanol/g consumed xylose. Among the selected hybrid strains, the hybrid FS22 showed the best fermentation capacity in 75% WSLQ...... the inhibitor tolerance of this yeast, a combination of UV mutagenesis and protoplast fusion was used to construct strains with improved performance. Firstly, UVinduced mutants were screened and selected for improved tolerance towards furfural. The most promised mutant, S. passalidarum M7, produced 50% more...

Induction and catabolite repression of α-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis

NARCIS (Netherlands)

Wijk, R. van; Ouwehand, J.; Bos, T. van den; Koningsberger, V.V.

1969-01-01

1. 1. Kinetic data on the repression, the derepression and the induction of α-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis suggested that some site other than the stereospecific site for the induction by maltose was involved in the repression by glucose. 2. 2. A study of the

The effectiveness of shin guards used by football players.

Science.gov (United States)

Tatar, Yasar; Ramazanoglu, Nusret; Camliguney, Asiye Filiz; Saygi, Evrim Karadag; Cotuk, Hasan Birol

2014-01-01

In football, injuries from opponent contact occur commonly in the lower extremities. FIFA the world's governing body for football requires players to wear shin guards. The aim of this study was to compare the protective effectiveness of polypropylene based shin guards with custom-made carbon fiber ones. Three commercial polypropylene shin guards (Adidas Predator™, Adidas UCL™, and Nike Mercurial™) and two custom-made carbon fiber shin guards were examined. The experimental setup had the following parts: 1) A pendulum attached a load cell at the tip (CAS Corp., Korea) and a fixed prosthetic foot equipped with a cleat to simulate an attacker's foot. 2) An artificial tibia prepared by condensed foam and reinforced by carbon fibers protected with soft clothing. 3) A multifunctional sensor system (Tekscan Corp., F-Socket System, Turkey) to record the impact on the tibia. In the low impact force trials, only 2.79-9.63 % of the load was transmitted to the sensors. When comparing for mean force, peak force and impulse, both carbon fiber shin guards performed better than the commercial ones (Adidas Predator™, Adidas UCL™, and Nike Mercurial™) (p = 0.000). Based on these same parameters, the Nike Mercurial™ provided better protection than the Adidas Predator™ and the Adidas UCL™ (p = 0.000). In the high impact force trials, only 5.16-10.90 % of the load was transmitted to the sensors. For peak force and impulse, the carbon fiber shin guards provided better protection than all the others. Carbon fiber shin guards possess protective qualities superior to those of commercial polypropylene shin guards. Key PointsShin guards decrease the risk of serious injuries.Carbon shin guards provide sufficient protection against high impact forces.Commercially available Polypropylene based shin guards do not provide sufficient protection against high impact forces.

Produção de protoplastos e lise da parede celular de leveduras utilizando β-1,3 glucanase Protoplasts production and yeast cell wall lysis using β-1,3 glucanase

Directory of Open Access Journals (Sweden)

Luciana Francisco Fleuri

2010-06-01

Full Text Available O presente trabalho visou a aplicação da β-1,3 glucanase lítica, obtida do microrganismo Cellulosimicrobium cellulans 191, na produção de protoplastos e na lise da parede celular de leveduras. A preparação bruta da enzima foi capaz de lisar as leveduras Kluyveromyces lodderi, Saccharomyces cerevisiae (Fleischmann e Itaiquara, S. cerevisiae KL-88, S. diastaticus NCYC 713, S. cerevisiae NCYC 1001, Candida glabrata NCYC 388, Kluyveromyces marxianus NCYC 587 e Hansenula mrakii NCYC 500. A β-1,3 glucanase purificada foi capaz de lisar as leveduras Saccharomyces cerevisiae KL-88, Saccharomyces capensis, Debaromyces vanriji, Pachysolen tannophillus, Kluyveromyces drosophilarum, Candida glabrata, Hansenula mrakii e Pichia membranaefaciens e formar protoplastos de Saccharomyces cerevisiae KL-88.The aim of this work was the application of lytic β-1,3 glucanase obtained from Cellulosimicrobium cellulans strain 191 in the production of protoplasts and lysis of yeast cell walls. The crude extract demonstrated lysis activity against the yeasts Kluyveromyces lodderi, Saccharomyces cerevisiae (Fleischmann and Itaiquara, S. cerevisiae KL-88, S. diastaticus NCYC 713, S. cerevisiae NCYC 1001, Candida glabrata NCYC 388, Kluyveromyces marxianus NCYC 587, and Hansenula mrakii NCYC 500. The purified β-1,3 glucanase demonstrated lysis activity against the yeasts Saccharomyces cerevisiae KL-88, Saccharomyces capensis, Debaromyces vanriji, Pachysolen tannophillus, Kluyveromyces drosophilarum, Candida glabrata, Hansenula mrakii, and Pichia membranaefaciens, and it was able to produce Saccharomyces cerevisiae KL-88 protoplasts.

Synthesis of viral DNA forms in Nicotiana plumbaginifolia protoplasts inoculated with cassava latent virus (CLV); evidence for the independent replication of one component of the CLV genome.

OpenAIRE

Townsend, R; Watts, J; Stanley, J

1986-01-01

Totipotent leaf mesophyll protoplasts of Nicotiana plumbaginifolia, Viviani were inoculated with cassava latent virus (CLV) or with full length copies of CLV genomic DNAs 1 and 2 excised from replicative forms of M13 clones. Virus specific DNAs began to appear 48-72h after inoculation with virus or cloned DNAs, coincident with the onset of host cell division. Infected cells accumulated supercoiled forms of DNAs 1 and 2 as well as progeny single-stranded (ss) virion (+) sense DNAs representing...

NO and H2O2 contribute to SO2 toxicity via Ca2+ signaling in Vicia faba guard cells.

Science.gov (United States)

Yi, Min; Bai, Heli; Xue, Meizhao; Yi, Huilan

2017-04-01

NO and H 2 O 2 have been implicated as important signals in biotic and abiotic stress responses of plants to the environment. Previously, we have shown that SO 2 exposure increased the levels of NO and H 2 O 2 in plant cells. We hypothesize that, as signaling molecules, NO and H 2 O 2 mediate SO 2 -caused toxicity. In this paper, we show that SO 2 hydrates caused guard cell death in a concentration-dependent manner in the concentration range of 0.25 to 6 mmol L -1 , which was associated with elevation of intracellular NO, H 2 O 2 , and Ca 2+ levels in Vicia faba guard cells. NO donor SNP enhanced SO 2 toxicity, while NO scavenger c-PTIO and NO synthesis inhibitors L-NAME and tungstate significantly prevented SO 2 toxicity. ROS scavenger ascorbic acid (AsA) and catalase (CAT), Ca 2+ chelating agent EGTA, and Ca 2+ channel inhibitor LaCl 3 also markedly blocked SO 2 toxicity. In addition, both c-PTIO and AsA could completely block SO 2 -induced elevation of intracellular Ca 2+ level. Moreover, c-PTIO efficiently blocked SO 2 -induced H 2 O 2 elevation, and AsA significantly blocked SO 2 -induced NO elevation. These results indicate that extra NO and H 2 O 2 are produced and accumulated in SO 2 -treated guard cells, which further activate Ca 2+ signaling to mediate SO 2 toxicity. Our findings suggest that both NO and H 2 O 2 contribute to SO 2 toxicity via Ca 2+ signaling.

Plant, cell, and molecular mechanisms of abscisic-acid regulation of stomatal apertures. A new mechanism for the regulation of stomatal-aperture size in intact leaves: Accumulation of mesophyll-derived sucrose in the guard-cell wall of Vicia faba L.

Energy Technology Data Exchange (ETDEWEB)

Lu, P.; Outlaw, W.H. Jr.; Smith, B.G.; Freed, G.A.

1996-12-31

At various times after pulse labeling Vicia faba L. leaflets with {sup 14}CO{sub 2}, whole-leaf pieces and rinsed epidermal peels were harvested and subsequently processed for histochemical analysis. Cells dissected from whole leaf retained apoplastic contents whereas those from rinsed peels contained only cytoplastic contents. Sucrose specific radioactivity peaked in palisade cells, 111 GBq{center_dot}mol{sup {minus}1}, at 20 min. In contrast, the {sup 14}C content and sucrose specific radioactivity were very low in guard cells for 20 min, implying little CO{sub 2} incorporation; both then peaked at 40 min. The guard-cell apoplast had a high maximum sucrose specific radioactivity and a high sucrose influx rate. These and other comparisons implied the presence of (a) multiple sucrose pools in mesophyll cells, (b) a localized mesophyll-apoplast region that exchanges with phloem and stomata, and (c) mesophyll-derived sucrose in guard-cell walls sufficient to diminish stomatal opening by {approximately} 4 {micro}m. Factors expected to enhance sucrose accumulation in guard-cell walls are (a) high transpiration rate, which closes stomata, and (b) high apoplastic sucrose concentration, which is elevated when mesophyll-sucrose efflux exceeds translocation. Therefore, multiple physiological factors are integrated in the attenuation of stomatal-aperture size by this previously unrecognized mechanism.

C-terminus-mediated voltage gating of Arabidopsis guard cell anion channel QUAC1.

Science.gov (United States)

Mumm, Patrick; Imes, Dennis; Martinoia, Enrico; Al-Rasheid, Khaled A S; Geiger, Dietmar; Marten, Irene; Hedrich, Rainer

2013-09-01

Anion transporters in plants play a fundamental role in volume regulation and signaling. Currently, two plasma membrane-located anion channel families—SLAC/SLAH and ALMT—are known. Among the ALMT family, the root-expressed ALuminium-activated Malate Transporter 1 was identified by comparison of aluminum-tolerant and Al(3+)-sensitive wheat cultivars and was subsequently shown to mediate voltage-independent malate currents. In contrast, ALMT12/QUAC1 (QUickly activating Anion Channel1) is expressed in guard cells transporting malate in an Al(3+)-insensitive and highly voltage-dependent manner. So far, no information is available about the structure and mechanism of voltage-dependent gating with the QUAC1 channel protein. Here, we analyzed gating of QUAC1-type currents in the plasma membrane of guard cells and QUAC1-expressing oocytes revealing similar voltage dependencies and activation–deactivation kinetics. In the heterologous expression system, QUAC1 was electrophysiologically characterized at increasing extra- and intracellular malate concentrations. Thereby, malate additively stimulated the voltage-dependent QUAC1 activity. In search of structural determinants of the gating process, we could not identify transmembrane domains common for voltage-sensitive channels. However, site-directed mutations and deletions at the C-terminus of QUAC1 resulted in altered voltage-dependent channel activity. Interestingly, the replacement of a single glutamate residue, which is conserved in ALMT channels from different clades, by an alanine disrupted QUAC1 activity. Together with C- and N-terminal tagging, these results indicate that the cytosolic C-terminus is involved in the voltage-dependent gating mechanism of QUAC1.

Bird guard

Science.gov (United States)

Fairchild, Dana M [Armour, SD

2010-03-02

The bird guard provides a device to protect electrical insulators comprising a central shaft; a clamp attached to an end of the shaft to secure the device to a transmission tower; a top and bottom cover to shield transmission tower insulators; and bearings to allow the guard to rotate in order to frighten birds away from the insulators.

Irrational Guards are Sometimes Needed

DEFF Research Database (Denmark)

Abrahamsen, Mikkel; Adamaszek, Anna; Miltzow, Tillmann

2017-01-01

In this paper we study the art gallery problem, which is one of the fundamental problems in computational geometry. The objective is to place a minimum number of guards inside a simple polygon so that the guards together can see the whole polygon. We say that a guard at position x sees a point y...... if the line segment xy is contained in the polygon. Despite an extensive study of the art gallery problem, it remained an open question whether there are polygons given by integer coordinates that require guard positions with irrational coordinates in any optimal solution. We give a positive answer...... to this question by constructing a monotone polygon with integer coordinates that can be guarded by three guards only when we allow to place the guards at points with irrational coordinates. Otherwise, four guards are needed. By extending this example, we show that for every n, there is a polygon which can...

Major benefits of guarding behavior in subsocial bees: implications for social evolution.

Science.gov (United States)

Mikát, Michael; Černá, Kateřina; Straka, Jakub

2016-10-01

Parental care is a behavior that increases the growth and survival of offspring, often at a cost to the parents' own survival and/or future reproduction. In this study, we focused on nest guarding, which is one of the most important types of extended parental care; we studied this behavior in two solitary bee species of the genus Ceratina with social ancestors. We performed the experiment of removing the laying female, who usually guards the nest after completing its provisioning, to test the effects of nest guarding on the offspring survival and nest fate. By dissecting natural nests, we found that Ceratina cucurbitina females always guarded their offspring until the offspring reached adulthood. In addition, the females of this species were able to crawl across the nest partitions and inspect the offspring in the brood cells. In contrast, several Ceratina chalybea females guarded their nests until the offspring reached adulthood, but others closed the nest entrance with a plug and deserted the nest. Nests with a low number of provisioned cells were more likely to be plugged and abandoned than nests with a higher number of cells. The female removal experiment had a significantly negative effect on offspring survival in both species. These nests frequently failed due to the attacks of natural enemies (e.g., ants, chalcidoid wasps, and other competing Ceratina bees). Increased offspring survival is the most important benefit of the guarding strategy. The abandonment of a potentially unsuccessful brood might constitute a benefit of the nest plugging behavior. The facultative nest desertion strategy is a derived behavior in the studied bees and constitutes an example of an evolutionary reduction in the extent of parental care.

Coast Guard

International Nuclear Information System (INIS)

1991-09-01

The 11-million gallon Exxon Valdez oil spill highlighted deficiencies in the nation's ability to contain and recover spilled oil. The Oil Pollution Act of 1990 represents a major effort by Congress to address these deficiencies and to clarify the roles and responsibilities of the private sector and the federal government in preventing, preparing for, and responding to oil spills. This report examines the Coast Guard's efforts to avoid unnecessary and wasteful duplication by coordinating with the private sector and others, including federal and state agencies, its plans to buy oil spill response equipment and the new responsibilities the act places on the private sector and the Coast Guard and if these responsibilities call for a shift in emphasis in Coast Guard oil spill response activities

The Effectiveness of Shin Guards Used by Football Players

Directory of Open Access Journals (Sweden)

Yasar Tatar

2013-03-01

Full Text Available In football, injuries from opponent contact occur commonly in the lower extremities. FIFA the world’s governing body for football requires players to wear shin guards. The aim of this study was to compare the protective effectiveness of polypropylene based shin guards with custom-made carbon fiber ones. Three commercial polypropylene shin guards (Adidas Predator™, Adidas UCL™, and Nike Mercurial™ and two custom-made carbon fiber shin guards were examined. The experimental setup had the following parts: 1 A pendulum attached a load cell at the tip (CAS Corp., Korea and a fixed prosthetic foot equipped with a cleat to simulate an attacker’s foot. 2 An artificial tibia prepared by condensed foam and reinforced by carbon fibers protected with soft clothing. 3 A multifunctional sensor system (Tekscan Corp., F-Socket System, Turkey to record the impact on the tibia. In the low impact force trials, only 2.79-9.63 % of the load was transmitted to the sensors. When comparing for mean force, peak force and impulse, both carbon fiber shin guards performed better than the commercial ones (Adidas Predator™, Adidas UCL™, and Nike Mercurial™ (p = 0.000. Based on these same parameters, the Nike Mercurial™ provided better protection than the Adidas Predator™ and the Adidas UCL™ (p = 0.000. In the high impact force trials, only 5.16-10.90 % of the load was transmitted to the sensors. For peak force and impulse, the carbon fiber shin guards provided better protection than all the others. Carbon fiber shin guards possess protective qualities superior to those of commercial polypropylene shin guards.

Guarded Cubical Type Theory

DEFF Research Database (Denmark)

Birkedal, Lars; Bizjak, Aleš; Clouston, Ranald

2016-01-01

This paper improves the treatment of equality in guarded dependent type theory (GDTT), by combining it with cubical type theory (CTT). GDTT is an extensional type theory with guarded recursive types, which are useful for building models of program logics, and for programming and reasoning...... with coinductive types. We wish to implement GDTT with decidable type-checking, while still supporting non-trivial equality proofs that reason about the extensions of guarded recursive constructions. CTT is a variation of Martin-L\\"of type theory in which the identity type is replaced by abstract paths between...... terms. CTT provides a computational interpretation of functional extensionality, is conjectured to have decidable type checking, and has an implemented type-checker. Our new type theory, called guarded cubical type theory, provides a computational interpretation of extensionality for guarded recursive...

Photosynthetic CO2 fixation in guard cells (GC)

International Nuclear Information System (INIS)

Gotow, K.; Taylor, S.; Zeiger, E.

1987-01-01

Recent studies indicate that carbon metabolism in GC is modulated by light quality. The fate of 14 CO 2 supplied to highly purified Vicia GC protoplasts irradiated with red light was investigated. The suspension was stirred at 25 0 C and dark-adapted for 5 min. After 5 min. in red light, 4.8 uCi of NaH 14 CO 3 was added (final concentration: 100 uM). Metabolism was quenched after 30 s with boiling ethanol. Anionic compounds were separated by 2D PC and TLC, and quantified. Rates of CO 2 fixation were 5- to 8-fold higher in the light. In the dark, malate and aspartate had 90% of the total label; in the light, 3-PGA, sugar monophosphates (SMP) and sugar diophosphates (SDP) had up to 60% of the label. Phosphates treatment and rechromatography of labelled SDP showed the presence of ribulose, a specific PCRP metabolite. In time-course experiments, labelled 3-PGA was detected within 5 s. With time, the percentage of label in 3-PGA decreased and that in SMP increased. The authors conclude that 3-PGA is a primary carboxylation product of the PCRP in GC and that the activity of the PCRP and PEP-carboxylase is metabolically regulated

Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy

NARCIS (Netherlands)

Vermeer, J.E.M.; van Munster, E.B.; Vischer, N.O.; Gadella, T.

2004-01-01

Multimode fluorescence resonance energy transfer (FRET) microscopy was applied to study the plasma membrane organization using different lipidated green fluorescent protein (GFP)-fusion proteins co-expressed in cowpea protoplasts. Cyan fluorescent protein (CFP) was fused to the hyper variable region

Central functions of bicarbonate in S-type anion channel activation and OST1 protein kinase in CO 2 signal transduction in guard cell

KAUST Repository

Xue, Shaowu; Hu, Honghong; Ries, Amber; Merilo, Ebe; Kollist, Hannes; Schroeder, Julian I

2011-01-01

Plants respond to elevated CO(2) via carbonic anhydrases that mediate stomatal closing, but little is known about the early signalling mechanisms following the initial CO(2) response. It remains unclear whether CO(2), HCO(3)(-) or a combination activates downstream signalling. Here, we demonstrate that bicarbonate functions as a small-molecule activator of SLAC1 anion channels in guard cells. Elevated intracellular [HCO(3)(-)](i) with low [CO(2)] and [H(+)] activated S-type anion currents, whereas low [HCO(3)(-)](i) at high [CO(2)] and [H(+)] did not. Bicarbonate enhanced the intracellular Ca(2+) sensitivity of S-type anion channel activation in wild-type and ht1-2 kinase mutant guard cells. ht1-2 mutant guard cells exhibited enhanced bicarbonate sensitivity of S-type anion channel activation. The OST1 protein kinase has been reported not to affect CO(2) signalling. Unexpectedly, OST1 loss-of-function alleles showed strongly impaired CO(2)-induced stomatal closing and HCO(3)(-) activation of anion channels. Moreover, PYR/RCAR abscisic acid (ABA) receptor mutants slowed but did not abolish CO(2)/HCO(3)(-) signalling, redefining the convergence point of CO(2) and ABA signalling. A new working model of the sequence of CO(2) signalling events in gas exchange regulation is presented.

Central functions of bicarbonate in S-type anion channel activation and OST1 protein kinase in CO 2 signal transduction in guard cell

KAUST Repository

Xue, Shaowu

2011-03-18

Plants respond to elevated CO(2) via carbonic anhydrases that mediate stomatal closing, but little is known about the early signalling mechanisms following the initial CO(2) response. It remains unclear whether CO(2), HCO(3)(-) or a combination activates downstream signalling. Here, we demonstrate that bicarbonate functions as a small-molecule activator of SLAC1 anion channels in guard cells. Elevated intracellular [HCO(3)(-)](i) with low [CO(2)] and [H(+)] activated S-type anion currents, whereas low [HCO(3)(-)](i) at high [CO(2)] and [H(+)] did not. Bicarbonate enhanced the intracellular Ca(2+) sensitivity of S-type anion channel activation in wild-type and ht1-2 kinase mutant guard cells. ht1-2 mutant guard cells exhibited enhanced bicarbonate sensitivity of S-type anion channel activation. The OST1 protein kinase has been reported not to affect CO(2) signalling. Unexpectedly, OST1 loss-of-function alleles showed strongly impaired CO(2)-induced stomatal closing and HCO(3)(-) activation of anion channels. Moreover, PYR/RCAR abscisic acid (ABA) receptor mutants slowed but did not abolish CO(2)/HCO(3)(-) signalling, redefining the convergence point of CO(2) and ABA signalling. A new working model of the sequence of CO(2) signalling events in gas exchange regulation is presented.

OBTENÇÃO DE PLANTAS DE LIMÃO CRAVO (Citrus limonia Osbeck E TANGERINA CLEÓPATRA (Citrus reshni Hort. A PARTIR DO CULTIVO DE PROTOPLASTOS DE SUSPENSÃO CELULAR PLANT REGENERATION OF 'RANGPUR' LIME (Citrus limonia Osbeck AND 'CLEÓPATRA' MANDARIN (Citrus reshni Hort. THROUGH PROTOPLASTS OF CELL SUSPENSION

Directory of Open Access Journals (Sweden)

Rodrigo Rocha Latado

1999-01-01

Full Text Available Este trabalho descreve uma metodologia para a regeneração de plantas de tangerina 'Cleópatra' e limão 'Cravo', a partir do cultivo de protoplastos de suspensão celular. Para tal, calos nucelares foram induzidos em meio contendo BAP e cultivados em meio sem reguladores de crescimento. Protoplastos foram isolados de suspensões celulares e cultivados em gotas de agarose, com densidade de 2 X 105 protoplastos.ml-1. O meio MT, contendo ácido giberélico e água de coco, foi eficiente na germinação de embriões somáticos. Os métodos de aclimatação de plantas testados apresentaram baixa eficiência. Como resultado final, 17 plantas adaptadas de tangerina e 8 de limão foram obtidas.The present research describes the regeneration of 'Cleópatra' mandarin and 'Rangpur' lime plants from cell suspension protoplasts. Nucelar calli were induced on a medium containing BAP and maintained on growth regulator free medium. Protoplasts were isolated from embryogenic suspension and plated at a concentration of 2 X 105 protoplasts.ml-1, on agarose droplets. The MT medium with gibberellic acid and coconut water was efficient to stimulate somatic embryo conversion. Rooted plants acclimation had low efficiency. Seventeen mandarin plants and eight lime plants were obtained.

CLIMATE CHANGE FUEL CELL PROGRAM UNITED STATES COAST GUARD AIR STATION CAPE COD BOURNE, MASSACHUSETTS

Energy Technology Data Exchange (ETDEWEB)

John K. Steckel Jr

2004-06-30

This report covers the first year of operation of a fuel cell power plant, installed by PPL Spectrum, Inc. (PPL) under contract with the United States Coast Guard (USCG), Research and Development Center (RDC). The fuel cell was installed at Air Station Cape Cod in Bourne, MA. The project had the support of the Massachusetts Technology Collaborative (MTC), the Department of Energy (DOE), and Keyspan Energy. PPL selected FuelCell Energy, Inc. (FCE) and its fuel cell model DFC{reg_sign}300 for the contract. Grant contributions were finalized and a contract between PPL and the USCG for the manufacture, installation, and first year's maintenance of the fuel cell was executed on September 24, 2001. As the prime contractor, PPL was responsible for all facets of the project. All the work was completed by PPL through various subcontracts, including the primary subcontract with FCE for the manufacture, delivery, and installation of the fuel cell. The manufacturing and design phases proceeded in a relatively timely manner for the first half of the project. However, during latter stages of manufacture and fuel cell testing, a variety of issues were encountered that ultimately resulted in several delivery delays, and a number of contract modifications. Final installation and field testing was completed in April and May 2003. Final acceptance of the fuel cell was completed on May 16, 2003. The fuel cell has operated successfully for more than one year. The unit achieved an availability rate of 96%, which exceeded expectations. The capacity factor was limited because the unit was set at 155 kW (versus a nameplate of 250 kW) due to the interconnection with the electric utility. There were 18 shutdowns during the first year and most were brief. The ability of this plant to operate in the island mode improved availability by 3 to 4%. Events that would normally be shutdowns were simply island mode events. The mean time between failure was calculated at 239 hours, or slightly

Early stage differentiation of thallus cells of Porphyra haitanensis (Rhodophyta)

Science.gov (United States)

Wang, Sujuan; Sun, Yunlong; Lu, Anming; Wang, Guangyuan

1987-09-01

The early stage differentiation of thallus cells of Porphyra haitanensis T. J. Chang et B. F. Zheng was studied. Protoplasts or single cells were isolated from the blades using enzyme mixture comprising 2% sea snail gut enzyme and 1% cellulase. The isolated protoplasts or single cells were incubated in the MES medium. The cell differentiations were examined under the microscope at intervals after incubation. Four types of cell differentiation, namely, normal, abnormal, carposporangial and spermatorangial, and rhizoidal types, were observed. Since normal cell differentiations occur mostly in small thalli 50 mm in length and middle portions of big thalli 200 mm in length, it is essential to select tissues from these two kinds of thalli essential for commercial production.

Voltammetric detection of phytochelatin transported across unmodified and protoplast modified model phospholipid membranes

Czech Academy of Sciences Publication Activity Database

Navrátil, Tomáš; Nováková, Kateřina; Josypčuk, Bohdan; Sokolová, Romana; Šestáková, Ivana

2016-01-01

Roč. 147, č. 1 (2016), s. 165-171 ISSN 0026-9247 R&D Projects: GA ČR(CZ) GAP208/12/1645 Grant - others:Rada Programu interní podpory projektů mezinárodní spolupráce AV ČR(CZ) M200401201 Program:M Institutional support: RVO:61388955 Keywords : Barley protoplasts * electrochemical impendance spectroscopy * mercury electrode Subject RIV: CG - Electrochemistry Impact factor: 1.282, year: 2016

Coast Guard Compass

Science.gov (United States)

looks on as Adm. Charles Ray thanks Adm. Chuck Michel for his service as the 30th vice commandant of the commandant Adm. Charles W. Ray relieved Adm. Charles D. Michel as vice commandant of the U.S. Coast Guard by . Following graduation and commissioning as an officer in the Coast Guard, Wright will be heading to the

Guarded Cubical Type Theory

DEFF Research Database (Denmark)

Birkedal, Lars; Bizjak, Aleš; Clouston, Ranald

2016-01-01

This paper improves the treatment of equality in guarded dependent type theory (GDTT), by combining it with cubical type theory (CTT). GDTT is an extensional type theory with guarded recursive types, which are useful for building models of program logics, and for programming and reasoning...... with coinductive types. We wish to implement GDTT with decidable type checking, while still supporting non-trivial equality proofs that reason about the extensions of guarded recursive constructions. CTT is a variation of Martin-L\\"of type theory in which the identity type is replaced by abstract paths between...... terms. CTT provides a computational interpretation of functional extensionality, enjoys canonicity for the natural numbers type, and is conjectured to support decidable type-checking. Our new type theory, guarded cubical type theory (GCTT), provides a computational interpretation of extensionality...

Penetration of UV-B radiation in foliage: evidence that the epidermis behaves as a non-uniform filter

International Nuclear Information System (INIS)

Day, T.A.; Martin, G.; Vogelmann, T.C.

1993-01-01

In some plants, particularly herbaceous species, a considerable proportion of incident ultraviolet-B radiation (UV-B, 280-320 nm) penetrates into the leaf mesophyll where it is potentially damaging to nucleic acids and the photosynthetic machinery. We used optical techniques to look at the spatial variation in UV-B penetration through the epidermis of foliage of two herbaceous species (Chenopodium album and Smilacina stellata) and a conifer (Picea pungens). Measurements of UV-B penetration in intact foliage with a fibre-optic microprobe revealed that 300 nm radiation reached 161±36μm (mean±SD) into leaves of C. album, 154±40μm in S. stellata and 17±2μm in P. pungens, with epidermal transmittance being 39±14%, 55±19% and 0%, respectively. A thin polymer film was developed which fluoresced blue when irradiated by UV-B. Fresh epidermal leaf peels were placed over the film and irradiated with UV-B, and microscopic examination of the film from below allowed us to determine the spatial pattern of UV-B penetration through the epidermis. In herbaceous species, film fluorescence below cell walls, but not epidermal and guard cell protoplasts indicated that UV-B transmittance was much greater through anticlinal cell wall regions than protoplasts. Ultraviolet-B transmittance through large areas of epidermal cells could be induced by plasmolysis. Epidermal transmittance was also relatively high through stomal pores (and what appear to be nuclei in Smilacina), but relatively low through stomatal guard cells. Results from the fluorescing film technique were substantiated by direct measurements of UV-B transmittance through epidermal peels with a fibre-optic microprobe run paradermally along the bottom or inner side of irradiated peels. In Smilacina, we estimate that UV-B epidermal transmittance was up to 90% through anticlinal cell wall regions, but <10% through protoplast areas. In contrast to herbaceous species, we did not detect any UV-B transmittance through the

78 FR 11676 - Notice of Inventory Completion: National Guard Bureau/A7AN, Air National Guard, Joint Base...

Science.gov (United States)

2013-02-19

... Inventory Completion: National Guard Bureau/A7AN, Air National Guard, Joint Base Andrews, MD AGENCY..., Joint Base Andrews, MD, has completed an inventory of human remains and associated funerary objects, in... associated funerary objects may contact National Guard Bureau, Air National Guard, Joint Base Andrews, MD...

Truck side guard specifications : recommended standard

Science.gov (United States)

2016-09-01

This document is intended to be used by (1) public or private medium/heavy-duty truck fleets considering adding side guards; (2) jurisdictions or customers that require side guards through policy or procurement; (3) manufacturers of side guards; and ...

Videos - The National Guard

Science.gov (United States)

Legislative Liaison Small Business Programs Social Media State Websites Videos Featured Videos On Every Front 2:17 Always Ready, Always There National Guard Bureau Diversity and Inclusion Play Button 1:04 National Guard Bureau Diversity and Inclusion The ChalleNGe Ep.5 [Graduation] Play Button 3:51 The

30 CFR 75.1722 - Mechanical equipment guards.

Science.gov (United States)

2010-07-01

... may cause injury to persons shall be guarded. (b) Guards at conveyor-drive, conveyor-head, and conveyor-tail pulleys shall extend a distance sufficient to prevent a person from reaching behind the guard and becoming caught between the belt and the pulley. (c) Except when testing the machinery, guards...

30 CFR 77.400 - Mechanical equipment guards.

Science.gov (United States)

2010-07-01

... shall be guarded. (b) Overhead belts shall be guarded if the whipping action from a broken line would be hazardous to persons below. (c) Guards at conveyor-drive, conveyor-head, and conveyor-tail pulleys shall... between the belt and the pulley. (d) Except when testing the machinery, guards shall be securely in place...

Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis I. The effect of ribonuclease on protein synthesis

NARCIS (Netherlands)

Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

1961-01-01

Ribonuclease was found to inhibit the protein synthesis in the naked yeast protoplast for nearly 100%. Even small concentrations (5 μg/ml) were found inhibitory. The cause of this inhibition can be attributed at least in part to a 90% inhibition of the respiration. Amino acid uptake was found to

An optimal guarding scheme for thermal conductivity measurement using a guarded cut-bar technique, part 1 experimental study

International Nuclear Information System (INIS)

Xing, Changhu

2014-01-01

In the guarded cut-bar technique, a guard surrounding the measured sample and reference (meter) bars is temperature controlled to carefully regulate heat losses from the sample and reference bars. Guarding is typically carried out by matching the temperature profiles between the guard and the test stack of sample and meter bars. Problems arise in matching the profiles, especially when the thermal conductivities of the meter bars and of the sample differ, as is usually the case. In a previous numerical study, the applied guarding condition (guard temperature profile) was found to be an important factor in measurement accuracy. Different from the linear-matched or isothermal schemes recommended in literature, the optimal guarding condition is dependent on the system geometry and thermal conductivity ratio of sample to meter bar. To validate the numerical results, an experimental study was performed to investigate the resulting error under different guarding conditions using stainless steel 304 as both the sample and meter bars. The optimal guarding condition was further verified on a certified reference material, pyroceram 9606, and 99.95% pure iron whose thermal conductivities are much smaller and much larger, respectively, than that of the stainless steel meter bars. Additionally, measurements are performed using three different inert gases to show the effect of the insulation effective thermal conductivity on measurement error, revealing low conductivity, argon gas, gives the lowest error sensitivity when deviating from the optimal condition. The result of this study provides a general guideline for the specific measurement method and for methods requiring optimal guarding or insulation

Directory - Social Media - The National Guard

Science.gov (United States)

Brigade 1st Battalion, 145th Armored Regiment Headquarters and Headquarters Company, 1st Battalion, 145th Florida Air National Guard Florida Guard History Camp Blanding Joint Training Center Wolfpack Company, 1st Force Georgia National Guard Joint Force Headquarters 116th Air Control Wing Flickr Georgia National

Transient GFP expression in Nicotiana plumbaginifolia suspension cells: the role of gene silencing, cell death and T-DNA loss.

Science.gov (United States)

Weld, R; Heinemann, J; Eady, C

2001-03-01

The transient nature of T-DNA expression was studied with a gfp reporter gene transferred to Nicotiana plumbaginifolia suspension cells from Agrobacterium tumefaciens. Individual GFP-expressing protoplasts were isolated after 4 days' co-cultivation. The protoplasts were cultured without selection and 4 weeks later the surviving proto-calluses were again screened for GFP expression. Of the proto-calluses initially expressing GFP, 50% had lost detectable GFP activity during the first 4 weeks of culture. Multiple T-DNA copies of the gfp gene were detected in 10 of 17 proto-calluses lacking visible GFP activity. The remaining 7 cell lines contained no gfp sequences. Our results confirm that transiently expressed T-DNAs can be lost during growth of somatic cells and demonstrate that transiently expressing cells frequently integrate multiple T-DNAs that become silenced. In cells competent for DNA uptake, cell death and gene silencing were more important barriers to the recovery of stably expressing transformants than lack of T-DNA integration.

33 CFR 23.10 - Coast Guard emblem.

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Coast Guard emblem. 23.10 Section... DISTINCTIVE MARKINGS FOR COAST GUARD VESSELS AND AIRCRAFT § 23.10 Coast Guard emblem. (a) The distinctive emblem of the Coast Guard shall be as follows: On a disc the shield of the Coat of Arms of the United...

Approximation algorithms for guarding holey polygons ...

African Journals Online (AJOL)

Guarding edges of polygons is a version of art gallery problem.The goal is finding the minimum number of guards to cover the edges of a polygon. This problem is NP-hard, and to our knowledge there are approximation algorithms just for simple polygons. In this paper we present two approximation algorithms for guarding ...

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts Híbridos somáticos assimétricos de citros produzidos pela fusão de protoplastos irradiados e tratados com iodoacetamida

Directory of Open Access Journals (Sweden)

Claudine Maria de Bona

2009-05-01

Full Text Available The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad. cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow. and 'Changsha' mandarin (C. reticulata Blanco and 'Murcott' tangor (C. reticulata x C. sinensis. Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L-1 iodoacetamide (IOA, and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP analysis of plantlet DNAsamples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L-1 IBA solution for 10 min.O objetivo deste trabalho foi produzir híbridos somáticos assimétricos de citros pela fusão de protoplastos irradiados com raios gama e protoplastos tratados com iodoacetamida. Protoplastos foram isolados de suspensões celulares embriogênicas de pomelo (Citrus paradisi Macfad., cultivares Ruby Red e Flame, de laranja doce (C. sinensis Osbeck 'Itaboraí', 'Natal', Valencia' e 'Succari', de tangerinas 'Satsuma' (C. unshiu Marcow. e 'Changsha' (C. reticulata Blanco e de tangor 'Murcott' (C. reticulata x C. sinensis

Cation selectivity of the plasma membrane of tobacco protoplasts in the electroporated state.

Science.gov (United States)

Wegner, Lars H

2013-08-01

Cation selectivity of the cellular membrane of tobacco culture cells (cell line 'bright yellow-2') exposed to pulsed electric fields in the millisecond range was investigated. The whole cell configuration of the patch clamp technique was established on protoplasts prepared from these cells. Ion selectivity of the electroporated membrane was investigated by measuring the reversal potential of currents passing through field-induced pores. To this end the membrane was hyper- or depolarized for 10ms (prepulse); subsequently the voltage was driven to opposite polarity at a constant rate (+40 or -40mV/ms, respectively). The experiment was started by polarizing the membrane to moderately negative or positive voltages (prepulse potential ±150mV) that would not induce pore formation. Subsequently, an extended voltage range was scanned in the porated state of the membrane (prepulse potential ±600mV). IV curves in the porated and the non-porated state (obtained at the same prepulse polarity) were superimposed to determine the voltage at which both curves intersected ('Intersection potential'). Using a modified version of the Goldmann-Hodgkin-Katz equation relative permeabilities to Ca(2+) and various monovalent alkali and organic cations were calculated. Pores were found to be fairly cation selective, with a selectivity sequence determined to be Ca(2+)>Li(+)>Rb(+)≈K(+)≈Na(+)>TEA(+)≈TBA(+)>Cl(-). Relative permeability to monovalent cations was inversely related to the ionic diameter. By fitting a formalism suggested by Dwyer at al. (J. Gen. Physiol. 75 (1980), 469-492) the effective average diameter of field induced pores was estimated to be about 1.8nm. Implications of these results for biotechnology and electroporation theory are discussed. Copyright © 2013 Elsevier B.V. All rights reserved.

Minimizing guard ring dead space in silicon detectors with an n-type guard ring at the edge of the detector

International Nuclear Information System (INIS)

Palviainen, Tanja; Tuuva, Tuure; Leinonen, Kari

2007-01-01

Detectors with n-type silicon with an n + -type guard ring were investigated. In the present work, a new p + /n/n + detector structure with an n + guard ring is described. The guard ring is placed at the edge of the detector. The detector depletion region extends also sideways, allowing for signal collection very close to the n-guard ring. In this kind of detector structure, the dead space of the detector is minimized to be only below the guard ring. This is proved by simulations done using Silvaco/ATLAS software

Minimizing guard ring dead space in silicon detectors with an n-type guard ring at the edge of the detector

Energy Technology Data Exchange (ETDEWEB)

Palviainen, Tanja [Lappeenranta University of Technology, P.O. Box 20, FIN-53851 Lappeenranta (Finland)]. E-mail: tanja.palviainen@lut.fi; Tuuva, Tuure [Lappeenranta University of Technology, P.O. Box 20, FIN-53851 Lappeenranta (Finland); Leinonen, Kari [Lappeenranta University of Technology, P.O. Box 20, FIN-53851 Lappeenranta (Finland)

2007-04-01

Detectors with n-type silicon with an n{sup +}-type guard ring were investigated. In the present work, a new p{sup +}/n/n{sup +} detector structure with an n{sup +} guard ring is described. The guard ring is placed at the edge of the detector. The detector depletion region extends also sideways, allowing for signal collection very close to the n-guard ring. In this kind of detector structure, the dead space of the detector is minimized to be only below the guard ring. This is proved by simulations done using Silvaco/ATLAS software.

Ca2+ transport in plant cells and mechanisms of transformation of phytochrome-induced photosignals

Science.gov (United States)

Volotovski, Igor D.

1995-01-01

The recent data on the influence of phytochrome on the efficiency of Ca2+ translocation across the membranes of oat protoplasts are given. Ca2+ uptake in the protoplasts was shown to be influenced by the red light (R) illumination. This effect was reverted by the following far-red light (FR) illumination. To elucidate the sensitivity to phytochrome-controlling action the screening between the mechanisms of Ca2+ transport across the plasma membranes of oat protoplasts, Na+/Ca2+ and Ca2+/H+ exchangers, Ca2+-pump and Ca2+-channel was done. It was established that phytochrome modulated the activity of Na+/Ca2+-exchanger and Ca2+-pump. The light-mediated oscillations of cytoplasmic Ca2+ concentration in the oat protoplasts were demonstrated using fluorescence probe quin2 loaded into the cells and laser monitoring of fluorescence signal. The evidences were obtained that the oscillations were not the result of the elevation of cytoplasmic Ca2+ concentration and had no connection with Ca2+ pool of mitochondria. The possibility of the relation between the Ca2+ oscillations and phosphoinositide metabolism in plant cell membranes is analyzed. The mechanisms of transformation of primary phytochrome signal into biological effects were discussed.

Enhancement of yellow pigment production by intraspecific protoplast fusion of Monascus spp. yellow mutant (ade(-)) and white mutant (prototroph).

Science.gov (United States)

Klinsupa, Worawan; Phansiri, Salak; Thongpradis, Panida; Yongsmith, Busaba; Pothiratana, Chetsada

2016-01-10

To breed industrially useful strains of a slow-growing, yellow pigment producing strain of Monascus sp., protoplasts of Monascus purpureus yellow mutant (ade(-)) and rapid-growing M. purpureus white mutant (prototroph) were fused and fusants were selected on minimal medium (MM). Preliminary conventional protoplast fusion of the two strains was performed and the result showed that only white colonies were detected on MM. It was not able to differentiate the fusants from the white parental prototroph. To solve this problem, the white parental prototroph was thus pretreated with 20mM iodoacetamide (IOA) for cytoplasm inactivation and subsequently taken into protoplast fusion with slow-growing Monascus yellow mutant. Under this development technique, only the fusants, with viable cytoplasm from Monascus yellow mutant (ade(-)), could thus grow on MM, whereas neither IOA pretreated white parental prototroph nor yellow auxotroph (ade(-)) could survive. Fifty-three fusants isolated from yellow colonies obtained through this developed technique were subsequently inoculated on complete medium (MY agar). Fifteen distinguished yellow colonies from their parental yellow mutant were then selected for biochemical, morphological and fermentative properties in cassava starch and soybean flour (SS) broth. Finally, three most stable fusants (F7, F10 and F43) were then selected and compared in rice solid culture. Enhancement of yellow pigment production over the parental yellow auxotroph was found in F7 and F10, while enhanced glucoamylase activity was found in F43. The formation of fusants was further confirmed by monacolin K content, which was intermediate between the two parents (monacolin K-producing yellow auxotroph and non-monacolin K producing white prototroph). Copyright © 2015 Elsevier B.V. All rights reserved.

46 CFR 50.10-25 - Coast Guard Symbol.

Science.gov (United States)

2010-10-01

... 46 Shipping 2 2010-10-01 2010-10-01 false Coast Guard Symbol. 50.10-25 Section 50.10-25 Shipping... Definition of Terms Used in This Subchapter § 50.10-25 Coast Guard Symbol. (a) The term Coast Guard Symbol... impression of the Coast Guard Symbol for stamping nameplates and specimens is shown in Figure 50.10-25(b...

Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis II. Reversal of the RNase effect of protein synthesis by polymethacrylic acid

NARCIS (Netherlands)

Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

1961-01-01

The ribonuclease inhibited protein synthesis and respiration of yeast protoplasts can be restored by the addition of several polyanionic compounds, among which polymethacrylic acid proved to be the most effective one. The results of preliminary experiments with the ultracentrifuge indicate a

Guarding America: Security Guards and U.S. Critical Infrastructure Protection

National Research Council Canada - National Science Library

Parfomak, Paul W

2004-01-01

The Bush Administration's 2003 National Strategy for the Physical Protection of Critical Infrastructures and Key Assets indicates that security guards are an important source of protection for critical facilities...

Volunteers in the Danish Home Guard 2016

DEFF Research Database (Denmark)

Fridberg, Torben; Larsen, Mona

This report maps the composition of a group of volunteer members of the Home Guard, as well as their opinions and expectations of the Home Guard and their own voluntary efforts. The report is a follow-up to two previous surveys completed in 2007 and 2011 and it therefore also highlights changes...... from 2007 to 2011 and 2016. Based on a questionnaire survey, the report paints a picture of who the volunteers are, what motivates them and how they perceive their surrounding environment’s view of them as members of the Home Guard. The report also focuses on the volunteers’ view of the Home Guard......’s tasks and activities both in Denmark and abroad. Finally, the report describes the volunteers’ perception of the Home Guards’ communication and campaigns. The report was commissioned and financed by the Danish Home Guard Command....

Fact Sheets - The National Guard

Science.gov (United States)

National Guard About Us By the Numbers Contact Us FAQ Federal Mission History Join Us Leaders Director of March Today in Guard History Leadership CNGB VCNGB SEA DANG DARNG Joint Staff J-1 J-2 J-3 J-4 J-5 J-6 J Marshal Office of the Joint Surgeon PARC Small Business Programs Chaplain Diversity NGB-GOMO Resources

Biosynthesis of acid phosphatase of baker's yeast . Characterization of a protoplast-bound fraction containing precursors of the exo-enzyme

NARCIS (Netherlands)

Boer, Pieter; Rijn, Herman J.M. van; Reinking, A.; Steyn-Parvé, Elizabeth P.

1975-01-01

1. 1.|Yest protoplasts, secreting acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum) EC 3.1.3.2) contain a small amount of firmly bound enzyme, even after lysis (Van Rijn, H.J.M.; Boer, P. and Steyn-Parvé, E.P. (1972) Biochim. Biophys. Acta 268, 431–441). The major part

Security Guards for the Future Web

National Research Council Canada - National Science Library

Reed, Nancy; Bryson, Dave; Garriss, James; Gosnell, Steve; Heaton, Brook; Huber, Gary; Jacobs, David; Pulvermacher, Mary; Semy, Salim; Smith, Chad; Standard, John

2004-01-01

.... Guard technology needs to keep pace with the evolving Web environment. The authors conjectured that a family of security guard services would be needed to provide the full range of functionality necessary to support the future Web...

QTL-mapping in mink (Neovison vison) shows evidence for QTL for guard hair thickness, guard hair length and skin length

DEFF Research Database (Denmark)

Thirstrup, Janne Pia; Labouriau, Rodrigo; Guldbrandtsen, Bernt

2011-01-01

Fur quality in mink (Neovison vison) is a composite trait, consisting of e.g. guard hair length, guard hair thickness and density of wool. A genome wide QTL search was performed to detect QTL for fur quality traits in mink. Here we present the results of QTL analyses for guard hair length, guard...... hair thickness and density of wool. Data from an F2-cross was analysed across fourteen chromosomes using 100 microsatellites as markers with a spacing of approximately 20 cM. The two lines used for the F2-cross were Nordic wild mink and American short nap mink. In total 1,083 animals (21 wild type, 25...... short nap, 103 F1 and 934 F2) were marker typed and recorded for the three presented fur quality traits. For the QTL-analyses a regression analysis implemented in QTL Express software was used. Evidence was found for the existence of QTL for guard hair length, guard hair thickness and density of wool...

Model-Based Analysis of Arabidopsis Leaf Epidermal Cells Reveals Distinct Division and Expansion Patterns for Pavement and Guard Cells1[W][OA

Science.gov (United States)

Asl, Leila Kheibarshekan; Dhondt, Stijn; Boudolf, Véronique; Beemster, Gerrit T.S.; Beeckman, Tom; Inzé, Dirk; Govaerts, Willy; De Veylder, Lieven

2011-01-01

To efficiently capture sunlight for photosynthesis, leaves typically develop into a flat and thin structure. This development is driven by cell division and expansion, but the individual contribution of these processes is currently unknown, mainly because of the experimental difficulties to disentangle them in a developing organ, due to their tight interconnection. To circumvent this problem, we built a mathematic model that describes the possible division patterns and expansion rates for individual epidermal cells. This model was used to fit experimental data on cell numbers and sizes obtained over time intervals of 1 d throughout the development of the first leaf pair of Arabidopsis (Arabidopsis thaliana). The parameters were obtained by a derivative-free optimization method that minimizes the differences between the predicted and experimentally observed cell size distributions. The model allowed us to calculate probabilities for a cell to divide into guard or pavement cells, the maximum size at which it can divide, and its average cell division and expansion rates at each point during the leaf developmental process. Surprisingly, average cell cycle duration remained constant throughout leaf development, whereas no evidence for a maximum cell size threshold for cell division of pavement cells was found. Furthermore, the model predicted that neighboring cells of different sizes within the epidermis expand at distinctly different relative rates, which could be verified by direct observations. We conclude that cell division seems to occur independently from the status of cell expansion, whereas the cell cycle might act as a timer rather than as a size-regulated machinery. PMID:21693673

Chief, National Guard Bureau - Leadership - The National Guard

Science.gov (United States)

the ARNG Deputy Director of the ARNG Chief of Staff of the ARNG Command Chief Warrant Officer of the the ANG Chief of Staff of the ARNG CCW Officer of the ARNG CSM of the ARNG Bottom Links National Guard Civic Leader's Guide ARNG Vision 2020 Posture Statement Strategic Direction CNGB ARNG Financial Report

The Lettuce infectious yellows virus (LIYV)-encoded P26 is associated with plasmalemma deposits within LIYV-infected cells

International Nuclear Information System (INIS)

Medina, V.; Sudarshana, M.R.; Tian, T.; Ralston, K.S.; Yeh, H.-H.; Falk, B.W.

2005-01-01

Cytological, immunological, and mutagenesis approaches were used to identify the viral factors associated with the formation of plasmalemma deposits (PLDs) in whole plants and protoplasts infected by Lettuce infectious yellows virus (LIYV). Transmission electron microscopy and immunogold labeling using polyclonal antibodies to four of the five LIYV RNA 2-encoded large proteins, capsid protein (CP), minor capsid protein (CPm), HSP70 homolog (HSP70h), and P59, showed specific labeling of LIYV virions or virion aggregates around the vesiculated membranous inclusions, but not PLDs in LIYV-infected Nicotiana benthamiana, Nicotiana clevelandii, Lactuca sativa, and Chenopodium murale plants, and Nicotiana tabacum protoplasts. In contrast, antibodies to the RNA 2-encoded P26 showed specific labeling of PLDs but not virions in both LIYV-infected plants and protoplasts. Virion-like particles (VLPs) were seen in protoplasts infected by all LIYV RNA 2 mutants except for the CP (major capsid protein) mutant. PLDs were more difficult to find in protoplasts, but were seen in protoplasts infected by the CP and CPm mutants, but not in protoplasts infected by the P26, HSP70h, or P59 mutants. Interestingly, although the CPm mutant showed VLPs and PLDs, the PLDs did not show associated virions/virion-like particles as was always observed for PLDs seen in protoplasts infected by wild-type LIYV. Immunoblot analyses performed on purified LIYV virions showed that P26 was not detected with purified virions, but was detected in the cell wall, 1000 g and 30,000 g pellet fractions of LIYV-infected plants. These data suggest that P26 is associated with the LIYV-induced PLDs, and in contrast to the other RNA 2-encoded large proteins, P26 is not a virion protein

46 CFR 108.223 - Guards on exposed equipment.

Science.gov (United States)

2010-10-01

... have hand covers, guards, or rails installed on all belts, gears, shafts, pulleys, sprockets, spindles... 46 Shipping 4 2010-10-01 2010-10-01 false Guards on exposed equipment. 108.223 Section 108.223 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) A-MOBILE OFFSHORE DRILLING UNITS DESIGN...

Functional identification of a GORK potassium channel from the ancient desert shrub Ammopiptanthus mongolicus (Maxim.) Cheng f.

Science.gov (United States)

Li, Junlin; Zhang, Huanchao; Lei, Han; Jin, Man; Yue, Guangzhen; Su, Yanhua

2016-04-01

A GORK homologue K(+) channel from the ancient desert shrub Ammopiptanthus mongolicus (Maxim.) Cheng f. shows the functional conservation of the GORK channels among plant species. Guard cell K(+) release through the outward potassium channels eventually enables the closure of stomata which consequently prevents plant water loss from severe transpiration. Early patch-clamp studies with the guard cells have revealed many details of such outward potassium currents. However, genes coding for these potassium-release channels have not been sufficiently characterized from species other than the model plant Arabidopsis thaliana. We report here the functional identification of a GORK (for Gated or Guard cell Outward Rectifying K(+) channels) homologue from the ancient desert shrub Ammopiptanthus mongolicus (Maxim.) Cheng f. AmGORK was primary expressed in shoots, where the transcripts were regulated by stress factors simulated by PEG, NaCl or ABA treatments. Patch-clamp measurements on isolated guard cell protoplasts revealed typical depolarization voltage gated outward K(+) currents sensitive to the extracelluar K(+) concentration and pH, resembling the fundamental properties previously described in other species. Two-electrode voltage-clamp analysis in Xenopus lavies oocytes with AmGORK reconstituted highly similar characteristics as assessed in the guard cells, supporting that the function of AmGORK is consistent with a crucial role in mediating stomatal closure in Ammopiptanthus mongolicus. Furthermore, a single amino acid mutation D297N of AmGORK eventually abolishes both the voltage-gating and its outward rectification and converts the channel into a leak-like channel, indicating strong involvement of this residue in the gating and voltage dependence of AmGORK. Our results obtained from this anciently originated plant support a strong functional conservation of the GORK channels among plant species and maybe also along the progress of revolution.

A model of PCF in guarded type theory

DEFF Research Database (Denmark)

Paviotti, Marco; Møgelberg, Rasmus Ejlers; Birkedal, Lars

2015-01-01

Guarded recursion is a form of recursion where recursive calls are guarded by delay modalities. Previous work has shown how guarded recursion is useful for constructing logics for reasoning about programming languages with advanced features, as well as for constructing and reasoning about element...... adequate. The model construction is related to Escardo's metric model for PCF, but here everything is carried out entirely in type theory with guarded recursion, including the formulation of the operational semantics, the model construction and the proof of adequacy...... of coinductive types. In this paper we investigate how type theory with guarded recursion can be used as a metalanguage for denotational semantics useful both for constructing models and for proving properties of these. We do this by constructing a fairly intensional model of PCF and proving it computationally...

A Model of PCF in Guarded Type Theory

DEFF Research Database (Denmark)

Paviotti, Marco; Møgelberg, Rasmus Ejlers; Birkedal, Lars

2015-01-01

Guarded recursion is a form of recursion where recursive calls are guarded by delay modalities. Previous work has shown how guarded recursion is useful for constructing logics for reasoning about programming languages with advanced features, as well as for constructing and reasoning about element...... adequate. The model construction is related to Escardo's metric model for PCF, but here everything is carried out entirely in type theory with guarded recursion, including the formulation of the operational semantics, the model construction and the proof of adequacy....... of coinductive types. In this paper we investigate how type theory with guarded recursion can be used as a metalanguage for denotational semantics useful both for constructing models and for proving properties of these. We do this by constructing a fairly intensional model of PCF and proving it computationally...

How We Began - About the Guard - The National Guard

Science.gov (United States)

Marshal Office of the Joint Surgeon PARC Small Business Programs Chaplain Diversity NGB-GOMO Resources Legislative Liaison Small Business Programs Social Media State Websites Videos Featured Videos On Every Front the organization date of the oldest Army National Guard units is based in law. The Militia Act of May

46 CFR 177.940 - Guards in vehicle spaces.

Science.gov (United States)

2010-10-01

... 46 Shipping 7 2010-10-01 2010-10-01 false Guards in vehicle spaces. 177.940 Section 177.940... TONS) CONSTRUCTION AND ARRANGEMENT Rails and Guards § 177.940 Guards in vehicle spaces. On a vessel authorized to carry one or more vehicles, suitable chains, cables, or other barriers must be installed at the...

46 CFR 116.940 - Guards in vehicle spaces.

Science.gov (United States)

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false Guards in vehicle spaces. 116.940 Section 116.940... ARRANGEMENT Rails and Guards § 116.940 Guards in vehicle spaces. On a vessel authorized to carry one or more vehicles, suitable chains, cables, or other barriers must be installed at the end of each vehicle runway...

Issues related to choosing a guard force structure

International Nuclear Information System (INIS)

Auerback, C.; Cusack, J.; Green, L.; Higinbotham, W.; Indusi, J.; Marcuse, W.; Weinstock, E.

1975-01-01

The establishment of a federal security force has been suggested as an additional step to protect nuclear material. The force would be given the responsibility for guarding nuclear facilities and shipments of nuclear materials. This paper summarizes a study to identify the issues that are relevant to choosing between the private guard forces that are presently employed by industry or a guard force under federal authority. To examine the issues which would be relevant, three possible types of security force structures were selected for comparison: a federal force under central authority, a private guard force employed by or under contract to a facility, and a private arrangement to employ local police officers similar to the arrangement for armed guards at airports. The issues were divided into seven categories: (1) Legal Considerations, (2) Liability, (3) Staffing and Operations, (4) Costs, (5) Relationship to Off-site Forces, (6) Management and Control, (7) Transportation

Exe-Guard Project

Energy Technology Data Exchange (ETDEWEB)

Smith, Rhett [Schweitzer Engineering Lab., Inc., Pullman, WA (United States); Marshall, Tim [Dominion Virginia Power, Richmond, VA (United States); Chavez, Adrian [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Bratus, Sergey [Dartmouth College, Hanover, NH (United States)

2016-01-30

The exe-Guard Project is an alliance between Dominion Virginia Power (DVP), Sandia National Laboratories (SNL), Dartmouth University, and Schweitzer Engineering Laboratories (SEL). SEL is primary recipient on this project. The exe-Guard project was selected for award under DE-FOA-0000359 with CFDA number 81.122 to address Topic Area of Interest 4: Hardened platforms and Systems. The exe-Guard project developed an antivirus solution for control system embedded devices to prevent the execution of unauthorized code and maintain settings and configuration integrity. This project created a white list antivirus solution for control systems capable of running on embedded Linux® operating systems. White list antivirus methods allow only credible programs to run through the use of digital signatures and hash functions. Once a system’s secure state is baselined, white list antivirus software denies deviations from that state because of the installation of malicious code as this changes hash results. Black list antivirus software has been effective in traditional IT environments but has negative implications for control systems. Black list antivirus uses pattern matching and behavioral analysis to identify system threats while relying on regular updates to the signature file and recurrent system scanning. Black list antivirus is vulnerable to zero day exploits which have not yet been incorporated into a signature file update. System scans hamper the performance of high availability applications, as revealed in NIST special publication 1058 which summarizes the impact of blacklist antivirus on control systems: Manual or “on-demand” scanning has a major effect on control processes in that they take CPU time needed by the control process (Sometimes close to 100% of CPU time). Minimizing the antivirus software throttle setting will reduce but not eliminate this effect. Signature updates can also take up to 100% of CPU time, but for a much shorter period than a typical

Issues related to choosing a guard force structure

International Nuclear Information System (INIS)

Averbach, C.; Cusack, J.; Green, L.; Higinbotham, W.; Indusi, J.; Marcuse, W.; Weinstock, E.

1975-01-01

The establishment of a federal security force has been suggested as an additional step to protect nuclear material. The force would be given the responsibility for guarding nuclear facilities and shipments of nuclear materials. A study to identify the issues that are relevant to choosing between the private guard forces that are presently employed by industry or a guard force under federal authority is summarized. To examine the issues which would be relevant, three possible types of security force structures were selected for comparison: a federal force under central authority, a private guard force employed by or under contract to a facility, and a private arrangement to employ local police officers similar to the arrangement for armed guards at airports. The issues were divided into seven categories: (1) legal considerations, (2) liability, (3) staffing and operations, (4) costs, (5) relationship to off-site forces, (6) management and control, and (7) transportation. (U.S.)

Variability in the Foraging Distribution and Diet of Cape Gannets between the Guard and Post-guard Phases of the Breeding Cycle

Directory of Open Access Journals (Sweden)

Jonathan A. Botha

2018-02-01

Full Text Available During breeding, seabirds are central place foragers and are sensitive to changes in local prey availability. As the breeding season progresses, foraging behavior and distribution is expected to change in response to possible changes in local prey availability. In addition, adult gender, and the increasing nutritional demands of a growing chick may also influence the foraging behavior of individuals. At present, relatively few studies have assessed the foraging behavior of adult birds during the late post-guard stages of chick rearing. Through a combination of GPS tracking and diet sampling we investigated the foraging distances, spatial distribution, and prey composition of adult Cape gannets (Morus capensis during the guard and post-guard stages of chick rearing. We found no clear evidence for consistent sex-specific differences in foraging distances and spatial distribution during the guard stage, although marginal differences in the location of core foraging areas during the post-guard stage were apparent. Results, however, revealed a clear increase in foraging range from the early guard to the late post-guard stage of chick rearing. During December the diet was comprised almost exclusively of anchovy (Engraulis encrasicolus, the proportion of which had decreased significantly in the diet by January. This was mirrored by a substantial increase in the proportion of saury (Scomberesox saurus. These results suggest that Cape gannets show flexibility in the foraging behavior and diet, which may be related to changes in the abundance and distribution of prey or may reflect changes in the energetic requirements of the growing offspring. This study provides the first assessment of Cape gannet foraging behavior and spatial distribution during the post-guard stage of chick rearing. The importance of considering intra-annual variability in foraging distribution when using seabird tracking data in trophic and marine spatial planning studies are

ABA signaling in guard cells entails a dynamic protein-protein interaction relay from the PYL-RCAR family receptors to ion channels.

Science.gov (United States)

Lee, Sung Chul; Lim, Chae Woo; Lan, Wenzhi; He, Kai; Luan, Sheng

2013-03-01

Plant hormone abscisic acid (ABA) serves as an integrator of environmental stresses such as drought to trigger stomatal closure by regulating specific ion channels in guard cells. We previously reported that SLAC1, an outward anion channel required for stomatal closure, was regulated via reversible protein phosphorylation events involving ABA signaling components, including protein phosphatase 2C members and a SnRK2-type kinase (OST1). In this study, we reconstituted the ABA signaling pathway as a protein-protein interaction relay from the PYL/RCAR-type receptors, to the PP2C-SnRK2 phosphatase-kinase pairs, to the ion channel SLAC1. The ABA receptors interacted with and inhibited PP2C phosphatase activity against the SnRK2-type kinase, releasing active SnRK2 kinase to phosphorylate, and activate the SLAC1 channel, leading to reduced guard cell turgor and stomatal closure. Both yeast two-hybrid and bimolecular fluorescence complementation assays were used to verify the interactions among the components in the pathway. These biochemical assays demonstrated activity modifications of phosphatases and kinases by their interaction partners. The SLAC1 channel activity was used as an endpoint readout for the strength of the signaling pathway, depending on the presence of different combinations of signaling components. Further study using transgenic plants overexpressing one of the ABA receptors demonstrated that changing the relative level of interacting partners would change ABA sensitivity.

Oracle Data Guard 11gR2 administration beginner's guide

CERN Document Server

Baransel, Emre

2013-01-01

Using real-world examples and hands-on tasks, Oracle Data Guard 11gR2 Administration Beginner's Guide will give you a solid foundation in Oracle Data Guard. It has been designed to teach you everything you need to know to successfully create and operate Data Guard environments with maximum flexibility, compatibility, and effectiveness.If you are an Oracle database administrator who wants to configure and administer Data Guard configurations, then ""Oracle Data Guard 11gR2 Administration Beginner's Guide"" is for you. With a basic understanding of Oracle database administration, you'll be able

Denotational semantics for guarded dependent type theory

DEFF Research Database (Denmark)

Bizjak, Aleš; Møgelberg, Rasmus Ejlers

2018-01-01

We present a new model of Guarded Dependent Type Theory (GDTT), a type theory with guarded recursion and multiple clocks in which one can program with, and reason about coinductive types. Productivity of recursively defined coinductive programs and proofs is encoded in types using guarded recursion......, crucial for programming with coinductive types, types must be interpreted as presheaves orthogonal to the object of clocks. In the case of dependent types, this translates to a unique lifting condition similar to the one found in homotopy theoretic models of type theory. Since the universes defined...... by inclusions of clock variable contexts commute on the nose with type operations on the universes....

Roles for National Guard Components: Current Thoughts and Future Possibilities

Science.gov (United States)

2000-04-01

Gen Russell C. Davis, chief, National Guard Bureau, remarks at the National Chamber of Commerce , Washington, D.C., June 1999. 5 Moskos and Burk, 176...Lt Gen Russell C. Davis, chief, National Guard Bureau, remarks at the National Chamber of Commerce , Washington, D.C., June 9 1999. 5 National Guard...National Guard Bureau, remarks to the National Chamber of Commerce , Washington, D.C., June 1999, n.p. On-line. Internet, 27 February 2000, available at

Laser mutagenesis and producing cellulase condition optimization of Trichoderma virid protoplast

International Nuclear Information System (INIS)

Chen Shuli; Zhang Qin; Han Jingjing; Lv Jiangtao; Wang Shilong; Yao Side

2009-01-01

The protoplast of Trichoderma virid CICC13038 was mutated using Nd:YAG laser of 266 nm light. And a high-cellulase producing strain JG13 was bred by screening with cellulose microcrystalline. Under the condition of 28 degree C, 180 rpm and 72 h of fermentation time, optimal conditions for the celluase ferment by orthogonal experiment were: 2% bran as the carbon source, 1% (NH 4 ) 2 SO 4 as the nitrogen source, 0.5% Tween-80 as a enzyme-promoting agent,and 25 mL of medium volume in a 250 mL bottle. The cellulase activity of the mutant reached 35.68 U/mL, 25.76% higher than that of the original strain under the same conditions. The mutant JG13 has a great potential in industrial production. And it also can be used as the original strain for further mutagenesis to get the strain of higher cellulase activity. (authors)

Elastic-plastic dynamic behavior of guard pipes due to sudden opening of longitudinal cracks in the inner pipe and crash to the guard pipe wall

International Nuclear Information System (INIS)

Theuer, E.; Heller, M.

1979-01-01

Integrity of guard pipes is an important parameter in the design of nuclear steam supply systems. A guard pipe shall withstand all kinds of postulated inner pipe breaks without failure. Sudden opening of a crack in the inner pipe and crash of crack borders to the guard pipe wall represent a shock problem where complex phenomena of dynamic plastification as well as dynamic behavior of the entire system have to be taken in consideration. The problem was analyzed by means of Finite Element computation using the general purpose program MARC. Equation of motion was resolved by direct integration using the Newmark β-operator. Analysis shows that after 1,2 m sec crack borders touch the guard pipe wall for the first time. At this moment a considerable amount of local plastification appears in the inner pipe wall, while the guard pipe is nearly unstressed. After initial touching, the crack borders begin to slip along the guard pipe wall. Subsequently, a short withdrawal of the crack borders and a new crash occur, while the inner pipe rolls along the guard pipe wall. The analysis procedure described is suitable for designing numerous guard pipe geometries as well as U-Bolt restraint systems which have to withstand high-energy pipe rupture impact. (orig.)

30 CFR 77.1011 - Drill holes; guarding.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Drill holes; guarding. 77.1011 Section 77.1011 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH... Control § 77.1011 Drill holes; guarding. Drill holes large enough to constitute a hazard shall be covered...

Plant cell engineering: current research, application and future prospects

International Nuclear Information System (INIS)

Wang Xunqing; Liu Luxiang

2008-01-01

This paper reviewed the current status of basic research in plant cell engineering, highlighted the application of embryo culture, double haploid (DH) technology, protoplast culture and somatic hybridization, somaclonal variation, rapid propagation, and bio-products production of plant-origin, and t he prospects. (authors)

41 CFR 50-204.5 - Machine guarding.

Science.gov (United States)

2010-07-01

... be such that it does not offer an accident hazard in itself. (c) Point of Operation Guarding. (1) Point of operation is the area on a machine where work is actually performed upon the material being... guarding device shall be so designed and constructed so as to prevent the operator from having any part of...

Introduction of transformed chloroplasts from tobacco into petunia by asymmetric cell fusion.

Science.gov (United States)

Sigeno, Asako; Hayashi, Sugane; Terachi, Toru; Yamagishi, Hiroshi

2009-11-01

Plastid engineering technique has been established only in Nicotiana tabacum, and the widespread application is severely limited so far. In order to exploit a method to transfer the genetically transformed plastomes already obtained in tobacco into other plant species, somatic cell fusion was conducted between a plastome transformant of tobacco and a cultivar of petunia (Petunia hybrida). A tobacco strain whose plastids had been transformed with aadA (a streptomycin/spectinomycin adenylyltransferase gene) and mdar [a gene for monodehydroascorbate reductase (MDAR)] and a petunia variety, 'Telstar', were used as cell fusion partners. An efficient regeneration system from the protoplasts of both the parents, and effectiveness of selection for the aadA gene with spectinomycin were established before the cell fusion. In addition, the influence of UV irradiation on the callus development from the protoplasts and shoot regeneration of tobacco was investigated. Protoplasts were cultured after cell fusion treatment with polyethylene glycol, and asymmetric somatic cybrids were selected using the aadA gene as a marker. Although many shoots of tobacco that had escaped the UV irradiation regenerated, several shoots possessing the morphology of petunia and the resistance to spectinomycin were obtained. Molecular analyses of the petunia type regenerants demonstrated that they had the nuclear and mitochondrial genomes derived from petunia besides the chloroplasts of tobacco transformed with aadA and mdar. Furthermore, it was ascertained that mdar was transcribed in the somatic cybrids. The results indicate the success in intergeneric transfer of transformed plastids of tobacco into petunia.

Building an FTP guard

Energy Technology Data Exchange (ETDEWEB)

Sands, P.D.

1998-08-01

Classified designs usually include lesser classified (including unclassified) components. An engineer working on such a design needs access to the various sub-designs at lower classification levels. For simplicity, the problem is presented with only two levels: high and low. If the low-classification component designs are stored in the high network, they become inaccessible to persons working on a low network. In order to keep the networks separate, the component designs may be duplicated in all networks, resulting in a synchronization problem. Alternatively, they may be stored in the low network and brought into the high network when needed. The latter solution results in the use of sneaker-net (copying the files from the low system to a tape and carrying the tape to a high system) or a file transfer guard. This paper shows how an FTP Guard was constructed and implemented without degrading the security of the underlying B3 platform. The paper then shows how the guard can be extended to an FTP proxy server or an HTTP proxy server. The extension is accomplished by allowing the high-side user to select among items that already exist on the low-side. No high-side data can be directly compromised by the extension, but a mechanism must be developed to handle the low-bandwidth covert channel that would be introduced by the application.

30 CFR 56.12023 - Guarding electrical connections and resistor grids.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Guarding electrical connections and resistor... MINES Electricity § 56.12023 Guarding electrical connections and resistor grids. Electrical connections and resistor grids that are difficult or impractical to insulate shall be guarded, unless protection...

Studies on protein synthesis by protoplasts of saccharomyces carlsbergensis III. Studies on the specificity and the mechanism of the action of ribonuclease on protein synthesis

NARCIS (Netherlands)

Kloet, S.R. de; Dam, G.J.W. van; Koningsberger, V.V.

1962-01-01

In this paper, the experimental results are presented of a continued study on the specificity and the mechanism of the inhibition by ribonuclease of protein synthesis in protoplasts of Saccharomyces carlsbergensis. By comparing the effects of native pancreatic ribonuclease with those of

Characterization of a light-controlled anion channel in the plasma membrane of mesophyll cells of pea

NARCIS (Netherlands)

Elzenga, J.T.M.; Volkenburgh Van, E

In leaf mesophyll cells of pea (Pisum sativum) light induces a transient depolarization that is at least partly due to an increased plasma membrane conductance for anions. Several channel types were identified in the plasma membrane of protoplasts from mesophyll cells using the patch-clamp

Armed guards on vessels : insurance and liability

Directory of Open Access Journals (Sweden)

Mišo Mudrić

2011-12-01

Full Text Available The Paper examines the insurance and liability issues resulting from the use of armed guards on board vessels. The study begins with an overview of the available data on key economic fi gures representing the projected overall annual costs of modern piracy. The focus is then shifted to the issue of public versus private security, where possible dangers of private-based security options are discussed in general. After explaining why the Somalia region deserves a closer attention when compared to other pirate-infested waters, a brief summary of the international effort to combat piracy threat is presented, followed by a structured overview of the use of private maritime security options in the maritime sector in general. One security option is the use of armed guards on board vessels. This option is explored both from the political (the acceptance by stakeholders and legal standpoint (legal issues arising from the use of armed guards. An important remedy for the shipping companies/ operators threatened by the piracy hazard is the existence of affordable and effective (specialized marine insurance. A study of available piracy insurance policies is presented, followed by an analysis of case law and other legal issues arising from piracy attacks, which could prove important when considering the legal implications of armed guards employment. Finally, a simplifi ed economic analysis of available security options is presented, followed by the final assessment of benefi ts derived from the use of armed guards.

The role of oxytocin and vasopressin in conditioned mate guarding behavior in the female rat.

Science.gov (United States)

Holley, Amanda; Bellevue, Shannon; Vosberg, Daniel; Wenzel, Kerstin; Roorda, Sieger; Pfaus, James G

2015-05-15

We have shown previously that female rats given their first copulatory experiences with the same male rat display mate guarding behavior in the presence of that male provided a female competitor is also present. Females given access to the familiar male show more Fos induction within regions of the brain that contain oxytocin (OT) and vasopressin (AVP) cell bodies, notably the supraoptic (SON) and paraventricular nuclei (PVN) relative to females given sexual experience with different males. The present experiments examined whether the Fos induction we previously observed within the SON and PVN occurred within OT and/or AVP neurons, and whether exogenous administration of OT or AVP prior to female rats first sexual experience could potentiate the acquisition of mate guarding behavior. Female rats that display conditioned mate guarding had significantly more double-labeled Fos/OT neurons in both SON and PVN, and significantly more Fos/AVP neurons in the PVN. Peripheral administration of OT or AVP prior to their first sexual experience with the familiar male facilitated different aspects of mate guarding: OT augmented affiliative behaviors and presenting responses whereas AVP augmented interference behavior. These results indicate that female rats' first experiences with sexual reward when paired with the same male induce changes to bonding networks in the brain. Moreover peripheral administration of OT or AVP during their first sexual experience can augment different aspects of mate guarding behavior. Copyright © 2015 Elsevier Inc. All rights reserved.

Guarded dependent type theory with coinductive types

DEFF Research Database (Denmark)

Bizjak, Aleš; Grathwohl, Hans Bugge; Clouston, Ranald

2016-01-01

We present guarded dependent type theory, gDTT, an extensional dependent type theory with a later' modality and clock quantifiers for programming and proving with guarded recursive and coinductive types. The later modality is used to ensure the productivity of recursive definitions in a modular......, type based, way. Clock quantifiers are used for controlled elimination of the later modality and for encoding coinductive types using guarded recursive types. Key to the development of gDTT are novel type and term formers involving what we call delayed substitutions’. These generalise the applicative...... functor rules for the later modality considered in earlier work, and are crucial for programming and proving with dependent types. We show soundness of the type theory with respect to a denotational model....

30 CFR 77.509 - Transformers; installation and guarding.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Transformers; installation and guarding. 77.509... COAL MINES Electrical Equipment-General § 77.509 Transformers; installation and guarding. (a... feet from any energized parts, casings, or wiring. (b) Transformer stations shall be enclosed to...

The cyclic nucleotide gated cation channel AtCNGC10 traffics from the ER via Golgi vesicles to the plasma membrane of Arabidopsis root and leaf cells

Directory of Open Access Journals (Sweden)

Andres Marilou A

2007-09-01

Full Text Available Abstract Background The cyclic nucleotide-gated ion channels (CNGCs maintain cation homeostasis essential for a wide range of physiological processes in plant cells. However, the precise subcellular locations and trafficking of these membrane proteins are poorly understood. This is further complicated by a general deficiency of information about targeting pathways of membrane proteins in plants. To investigate CNGC trafficking and localization, we have measured Atcngc5 and Atcngc10 expression in roots and leaves, analyzed AtCNGC10-GFP fusions transiently expressed in protoplasts, and conducted immunofluorescence labeling of protoplasts and immunoelectron microscopic analysis of high pressure frozen leaves and roots. Results AtCNGC10 mRNA and protein levels were 2.5-fold higher in roots than leaves, while AtCNGC5 mRNA and protein levels were nearly equal in these tissues. The AtCNGC10-EGFP fusion was targeted to the plasma membrane in leaf protoplasts, and lightly labeled several intracellular structures. Immunofluorescence microscopy with affinity purified CNGC-specific antisera indicated that AtCNGC5 and AtCNGC10 are present in the plasma membrane of protoplasts. Immunoelectron microscopy demonstrated that AtCNGC10 was associated with the plasma membrane of mesophyll, palisade parenchyma and epidermal cells of leaves, and the meristem, columella and cap cells of roots. AtCNCG10 was also observed in the endoplasmic reticulum and Golgi cisternae and vesicles of 50–150 nm in size. Patch clamp assays of an AtCNGC10-GFP fusion expressed in HEK293 cells measured significant cation currents. Conclusion AtCNGC5 and AtCNGC10 are plasma membrane proteins. We postulate that AtCNGC10 traffics from the endoplasmic reticulum via the Golgi apparatus and associated vesicles to the plasma membrane. The presence of the cation channel, AtCNGC10, in root cap meristem cells, cell plate, and gravity-sensing columella cells, combined with the previously reported

30 CFR 77.510 - Resistors; location and guarding.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Resistors; location and guarding. 77.510 Section 77.510 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE... COAL MINES Electrical Equipment-General § 77.510 Resistors; location and guarding. Resistors, heaters...

National Guard Forces in the Cyber Domain

Science.gov (United States)

2015-05-22

TITLE AND SUBTITLE National Guard Forces in the Cyber Domain 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S...Soldiers. Army Cyber Command (ARCYBER) commander, Lieutenant General Edward Cardon stated that Guard will begin to build combat power with...90 2014 Quadrennial Defense Review, 15. 91 Ibid. 92 Edward C. Cardon , "ARMY.MIL, The Official Homepage of the United

33 CFR 125.11 - Form of Coast Guard Port Security Card.

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Form of Coast Guard Port Security... WATERFRONT FACILITIES OR VESSELS § 125.11 Form of Coast Guard Port Security Card. The Coast Guard Port... data. ...

Laser-mediated perforation of plant cells

Science.gov (United States)

Wehner, Martin; Jacobs, Philipp; Esser, Dominik; Schinkel, Helga; Schillberg, Stefan

2007-07-01

The functional analysis of plant cells at the cellular and subcellular levels requires novel technologies for the directed manipulation of individual cells. Lasers are increasingly exploited for the manipulation of plant cells, enabling the study of biological processes on a subcellular scale including transformation to generate genetically modified plants. In our setup either a picosecond laser operating at 1064 nm wavelength or a continuous wave laser diode emitting at 405 nm are coupled into an inverse microscope. The beams are focused to a spot size of about 1.5 μm and the tobacco cell protoplasts are irradiated. Optoporation is achieved when targeting the laser focal spot at the outermost edge of the plasma membrane. In case of the picosecond laser a single pulse with energy of about 0.4 μJ was sufficient to perforate the plasma membrane enabling the uptake of dye or DNA from the surrounding medium into the cytosol. When the ultraviolet laser diode at a power level of 17 mW is employed an irradiation time of 200 - 500 milliseconds is necessary to enable the uptake of macromolecules. In the presence of an EYFP encoding plasmid with a C-terminal peroxisomal signal sequence in the surrounding medium transient transformation of tobacco protoplasts could be achieved in up to 2% of the optoporated cells. Single cell perforation using this novel optoporation method shows that isolated plant cells can be permeabilized without direct manipulation. This is a valuable procedure for cell-specific applications, particularly where the import of specific molecules into plant cells is required for functional analysis.

Use of FTA gene guard filter paper for the storage and transportation of tumor cells for molecular testing.

Science.gov (United States)

Dobbs, Larry J; Madigan, Merle N; Carter, Alexis B; Earls, Lori

2002-01-01

Efficient methods of storing tumor specimens for molecular testing are needed in the modern surgical pathology laboratory. The FTA Gene Guard system is a novel method for the collection and room temperature storage of blood samples for DNA testing. The method uses index card-sized filter papers that provide an ideal medium on which to store tumor specimens for DNA testing. To determine whether FTA filter paper can be used in the surgical pathology laboratory to store tumor cells for DNA testing. Cell suspensions were prepared from 60 surgical specimens, and DNA was extracted either immediately or after storage on FTA paper. The DNA extracted by each method was tested by polymerase chain reaction (PCR) for the beta-globin and interferon gamma genes, and the results were compared. Fifteen lymph node specimens stored on FTA paper were then tested for immunoglobulin heavy chain (IgH) gene rearrangement by PCR, and these results were compared with those obtained for immediately extracted DNA. University medical center. The DNA extracted from cells stored on FTA paper performed as well in the PCR as the freshly extracted DNA in nearly all cases (>95%). The results of tests for IgH gene rearrangements showed 100% concordance between the 2 methods of DNA extraction.Conclusion.-Cells from surgical specimens can be stored on FTA paper for extended lengths of time, and DNA can be extracted from these cells for PCR-based testing. FTA filter paper is a reliable medium for the storage and/or transport of tumor cells for PCR-based DNA analysis.

Security Guards and Counter-terrorism: Tourism and Gaps in Terrorism Prevention

Directory of Open Access Journals (Sweden)

Luke Howie

2014-11-01

Full Text Available Organisation operating in the tourism industry are high priority targets for terrorists. In the aftermath of the 9/11 attacks tourism destinations, hotels and modes of public transportation have regularly been targeted by terrorists seeking to convey their violent message. As such, leaders and managers in the tourism industry carefully plan their security and counter-terrorism responses, often involving the hiring of security guards. It is here that I believe a significant gap in counter-terrorism preparedness exists. I argue that protecting tourism destinations is only possible if consideration is given the effectiveness of security guards and understanding that their well-being will impact upon their ability to deliver security. I want to draw attention to the often ignored social role of security guards. On 9/11, 42 security guards died whilst helping save the lives of thousands. They performed their jobs admirably, despite being low-paid, under-appreciated workers. In this paper I explore the social role of security guards in the context of tourism security. By drawing on representations of security guards in popular culture and reports on the state of the security guard industry. I argue that the lack of attention on the quality and well-being of guards is a significant black-spot in tourism security and terrorism preparedness.

Women's Fertility Status Alters Other Women's Jealousy and Mate Guarding.

Science.gov (United States)

Hurst, Ashalee C; Alquist, Jessica L; Puts, David A

2017-02-01

Across three studies, we tested the hypothesis that women exhibit greater jealousy and mate guarding toward women who are in the high (vs. low) fertility phase of their cycle. Women who imagined their partner with a woman pictured at high fertility reported more jealousy than women who imagined their partner with a woman pictured at low fertility (Studies 1 and 2). A meta-analysis across studies manipulating fertility status of the pictured woman found a significant effect of fertility status on both jealousy and mate guarding. Women with attractive partners viewed fertile-phase women as less trustworthy, which led to increased mate guarding (Study 2). In Study 3, the closer women were to peak fertility, the more instances they reported of other women acting jealously and mate guarding toward them. These studies provide evidence that women selectively exhibit jealousy and mate guarding toward women who are near peak fertility.

Connectivity for underway Coast Guard patrol boats

OpenAIRE

Busch, Gregory C.

1997-01-01

Approved for public release; distribution is unlimited This thesis examines the US Coast Guard patrol boat's ability to effectively exchange operational data while underway. The patrol boat is currently unable to obtain tactical law enforcement information from the central Law Enforcement Information System 2 (LEIS 2) database while on patrol. LEIS 2 provides access to law enforcement information from Coast Guard, FBI, and state and local law enforcement agencies. Availability of this info...

Male songbirds provide indirect parental care by guarding females during incubation

Science.gov (United States)

Fedy, B.C.; Martin, T.E.

2009-01-01

Across many taxa, guarding of fertile mates is a widespread tactic that enhances paternity assurance. However, guarding of mates can also occur during the nonfertile period, and the fitness benefits of this behavior are unclear. Male songbirds, for example, sometimes guard nonfertile females during foraging recesses from incubation. We hypothesized that guarding postreproductive mates may have important, but unrecognized, benefits by enhancing female foraging efficiency, thereby increasing time spent incubating eggs. We tested the hypothesis in 2 songbird species by examining female behavior during natural and experimentally induced absences of males. Male absence caused increased vigilance in foraging females that decreased their efficiency and resulted in less time spent incubating eggs. Male guarding of nonfertile females can thus provide a previously unrecognized form of indirect parental care.

What is a "good" encoding of guarded choice?

DEFF Research Database (Denmark)

Nestmann, Uwe

2000-01-01

into the latter that preserves divergence-freedom and symmetries. This paper argues that there are nevertheless "good" encodings between these calculi. In detail, we present a series of encodings for languages with (1) input-guarded choice, (2) both input and output-guarded choice, and (3) mixed-guarded choice......, and investigate them with respect to compositionality and divergence-freedom. The first and second encoding satisfy all of the above criteria, but various "good" candidates for the third encoding-inspired by an existing distributed implementation-invalidate one or the other criterion, While essentially confirming...... Palamidessi's result, our study suggests that the combination of strong compositionality and divergence-freedom is too strong for more practical purposes. (C) 2000 Academic Press....

A short term NaCl exposure increases the Na+ conductance of outward-rectified cation currents in the pith cells of sweet pepper

NARCIS (Netherlands)

Blom, M.; Vogelzang, S.A.

2003-01-01

The regulatory role of pith cells in the stem in Na recirculation in sweet pepper was investigated by evaluating the transport characteristics of the plasma membrane of this cell type and comparison with those of root cells. Ion conductivity and Na permeability of the plasma membranes of protoplasts

Design and evaluation of a low thermal electromotive force guarded scanner for resistance measurements

Science.gov (United States)

Jarrett, Dean G.; Marshall, James A.; Marshall, Thomas A.; Dziuba, Ronald F.

1999-06-01

The design and testing of a low thermal electromotive force guarded scanner, developed to provide completely guarded switching when used with actively guarded resistance bridge networks, is described. The design provides a continuous guard circuit trace on the scanner circuit boards that surrounds the relay contacts and protects the measurement circuit from leakages to ground. Modification to the circuit boards and relays of the guarded scanner are explained. Several tests were developed to evaluate the guarding effectiveness, including isolating sections of the guard circuit to create a potential drop between the main and guard circuits. Calibration of standard resistors using the guarded scanner has shown relative differences less than 1×10-6, 30×10-6, and 150×10-6 for measurements made with and without the guarded scanner at nominal resistances of 1, 10, and 100 GΩ, respectively. The substitution method was used to significantly reduce the relative differences between channels to less than 0.5×10-6, 3×10-6, and 30×10-6 for nominal resistances of 1, 10, and 100 GΩ, respectively. Applications for the guarded scanner in automated direct current measurement systems are presented.

Fricção em braquetes gerada por fios de aço inoxidável, superelásticos com IonGuard e sem IonGuard Friction force on brackets generated by stainless steel wire and superelastic wires with and without IonGuard

Directory of Open Access Journals (Sweden)

Luiz Carlos Campos Braga

2011-08-01

Full Text Available OBJETIVO: o objetivo deste estudo foi verificar a fricção no braquete (Roth, Composite, 10.17.005, 3,2mm, largura 0,022" x 0,030", Torque -2° e angulação +13°, Morelli®, Brasil, utilizando fios ortodônticos retangulares de 0,019" x 0,025" de aço inoxidável (Morelli®, Brasil e de níquel-titânio superelásticos Bioforce com IonGuard e sem IonGuard (Bioforce, GAC®, EUA. MÉTODOS: foram utilizados 24 conjuntos braquetes/segmento de fio, divididos em 3 grupos de acordo com o fio. Cada conjunto braquete/segmento de fio foi testado 3 vezes e obtida uma média. Os ensaios foram realizados em máquina universal de ensaios EMIC DL2000®. Os dados foram submetidos à Análise de Variância com significância de 95%. RESULTADOS: o fio retangular Bioforce com IonGuard apresentou fricção significativamente menor que o Bioforce sem IonGuard, porém sem diferença do fio de aço inoxidável. Entretanto, o coeficiente de variação dos fios Bioforce com e sem IonGuard foi menor que o do fio de aço inoxidável. CONCLUSÃO: os fios retangulares de 0,019" x 0,025" Bioforce com IonGuard apresentam menor fricção que o fio Bioforce sem IonGuard, sem diferença para o fio de aço inoxidável.OBJECTIVE: The aim of this study was to evaluate the friction forces on brackets (Roth, Composite, 10.17.005, 3.2 mm, width 0.022" x 0.030 ", Torque -2° and angulation +13°, Morelli®, Brazil, with stainless steel orthodontic rectangular wire (Morelli®, Brazil and nickel titanium superelastic Bioforce wires with and without IonGuard (Bioforce, GAC®, USA. MATERIAL AND METHODS: Twenty-four brackets/wire segment combinations were used, distributed into three groups according to the orthodontic wire. Each bracket/wire segment combination was tested three times. The tests were performed in a universal testing machine Emic DL2000®. The data was submitted to ANOVA one way followed by Tukey's post hoc test (p<0.05. RESULTS: The rectangular orthodontic Bioforce wire

Kinetics of Ca2+- and ATP-dependent, voltage-controlled anion conductance in the plasma membrane of mesophyll cells of Pisum sativum

NARCIS (Netherlands)

Elzenga, J.T.M.; van Volkenburgh, E.

Whole-cell patch-clamp techniques were used to measure anion currents through the plasma membrane of protoplasts of mesophyll cells of expanding pea (Pisum sativum L.) leaves. Voltage-induced changes of the currents could be modelled with single exponential activation and deactivation kinetics. The

Integrated Unit Deployments: Rethinking Air National Guard Fighter Mobilizations

Science.gov (United States)

2016-06-01

participation in the homeland defense mission. But they emphasized that homeland defense would not become an exclusive ANG mission. Operation...station that can absorb pilots and maintainers left behind. Few Air National Guard squadrons enjoy this luxury . Only Air Guard squadrons that are part

Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

Science.gov (United States)

Li, Jian-Feng; Bush, Jenifer; Xiong, Yan; Li, Lei; McCormack, Matthew

2011-01-01

Protein-protein interactions (PPIs) constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC) as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs) and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

Directory of Open Access Journals (Sweden)

Jian-Feng Li

Full Text Available Protein-protein interactions (PPIs constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

US Coast Guard Stations in Louisiana, Geographic NAD83, USCG [coast_guard_stations_USCG_1997

Data.gov (United States)

Louisiana Geographic Information Center — This is is a point dataset for the locations and attributes of eight US Coast Guard stations in Louisiana. The attributes include name, address, latitude (NAD27),...

The thiol compounds glutathione and homoglutathione differentially affect cell development in alfalfa (Medicago sativa L.).

Science.gov (United States)

Pasternak, Taras; Asard, Han; Potters, Geert; Jansen, Marcel A K

2014-01-01

Glutathione (GSH) is an important scavenger of Reactive Oxygen Species (ROS), precursor of metal chelating phytochelatins, xenobiotic defence compound and regulator of cell proliferation. Homoglutathione (hGSH) is a GSH homologue that is present in several taxa in the family of Fabaceae. It is thought that hGSH performs many of the stress-defence roles typically ascribed to GSH, yet little is known about the potential involvement of hGSH in controlling cell proliferation. Here we show that hGSH/GSH ratios vary across organs and cells and that these changes in hGSH/GSH ratio occur during dedifferentiation and/or cell cycle activation events. The use of a GSH/hGSH biosynthesis inhibitor resulted in impaired cytokinesis in isolated protoplasts, showing the critical importance of these thiol-compounds for cell division. However, exposure of isolated protoplasts to exogenous GSH accelerated cytokinesis, while exogenous hGSH was found to inhibit the same process. We conclude that GSH and hGSH have distinct functional roles in cell cycle regulation in Medicago sativa L. GSH is associated with meristemic cells, and promotes cell cycle activation and induction of somatic embryogenesis, while hGSH is associated with differentiated cells and embryo proliferation. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Volunteers in the Danish Home Guard 2011

DEFF Research Database (Denmark)

Fridberg, Torben; Damgaard, Malene

voluntary work than the population as a whole. The report also shows that one in three active members of the Home Guard would like to be deployed on international operations to support the armed forces. The young members are especially willing – and these members have increased in recent years. This report......This report describes the composition of the Home Guard’s volunteer members and their attitudes to and expectations for the Home Guard. A similar survey was carried out in 2007, and the present report therefore also examines the trends from 2007 to 2011. Among other things, the report shows...... that the voluntary members are a stable resource, as on average they have been members of the Home Guard for more than 24 years. There is a clear majority of men aged 25-50. Relatively many have vocational training, and many are employed in the private sector. Members are also relatively more active in other...

Use of Head Guards in AIBA Boxing Tournaments-A Cross-Sectional Observational Study.

Science.gov (United States)

Loosemore, Michael P; Butler, Charles F; Khadri, Abdelhamid; McDonagh, David; Patel, Vimal A; Bailes, Julian E

2017-01-01

This study looks at the changes in injuries after the implementation of a new rule by the International Boxing Association (AIBA) to remove head guards from its competitions. A cross-sectional observational study performed prospectively. This brief report examines the removal of head guards in 2 different ways. The first was to examine the stoppages due to blows to the head by comparing World Series Boxing (WSB), without head guards, to other AIBA competitions with head guards. Secondly, we examined the last 3 world championships: 2009 and 2011 (with head guards) and 2013 (without head guards). World Series Boxing and AIBA world championship boxing. Boxers from WSB and AIBA world championships. The information was recorded by ringside medical physicians. Stoppages per 10 000 rounds; stoppages per 1000 hours. Both studies show that the number of stoppages due to head blows was significantly decreased without head guards. The studies also showed that there was a notable increase in cuts. Removing head guards may reduce the already small risk of acute brain injury in amateur boxing.

The effect of the guard location in a nuclear facility on performance criteria

International Nuclear Information System (INIS)

Lee, Hyun-Chul; Hwang, In-Koo; An, Jin-Soo

2002-01-01

Full text: The major functions of a physical protection system (PPS) are detection, delay, and response. Detection function is mainly to sense an adversary's intrusion and assess alarm events, and its physical implementation includes intrusion sensors, alarm communication systems and entry control facilities. Delay function can be accomplished through various passive and active barriers, makes adversary's advance toward his goal slow. Response function is to practically stop adversary actions. There are response guards responsible for the response function in a nuclear facility. They can be located at one or more posts to take action against adversary's progress after alarm occurred. The mostly used quantitative effectiveness measure of a PPS is the probability of interruption (PI) derived from delay time after detection and guard response time. Delay time is a variable dependent on adversary's capabilities and guard response time varies with the distance between a guard post and destination to be dispatched. According to the advanced technology of digital computing, software programs such as SAVI (System Analysis of Vulnerability to Intrusion) provide PI based on user input including adversary capability, protective elements, guard response time, and other PPS information. However guard response time is considered as a constant not a variable. That means same guard response time is always set regardless of the variable guard travel time from a post to a location where guards can interrupt the adversary's action. Since the location of guards may be easily changed in contrast to detection elements or delay elements and can be considered as a flexible mean against temporarily upgraded threat, a trial to investigate the changes of PI and CDP (Critical Detection Point) in case that guard response time is a variable was started. We defined several detection and delay elements along an adversary intrusion path and guard travel times as many as the number of detection

The Impact of Excluding Food Guarding from a Standardized Behavioral Canine Assessment in Animal Shelters

Science.gov (United States)

Mohan-Gibbons, Heather; Dolan, Emily D.; Reid, Pamela; Slater, Margaret R.; Mulligan, Hugh; Weiss, Emily

2018-01-01

Simple Summary Recent research has called into question the value of the food guarding assessment as a predictive tool for determining the safety of shelter dogs. This study examined the effect of eliminating the food guarding assessment in nine U.S. animal shelters. It was found that when the food guarding assessment was removed, bites or other injuries to staff or adopters did not increase. However, dogs exhibiting food guarding behavior were less likely to be adopted, had a longer shelter stay, and were more likely to be euthanized than dogs in the general population. Based on previous research and this study’s findings, the authors recommend that shelters discontinue the food guarding assessment. Abstract Many shelters euthanize or restrict adoptions for dogs that exhibit food guarding while in the animal shelter. However, previous research showed that only half the dogs exhibiting food guarding during an assessment food guard in the home. So, dogs are often misidentified as future food guarders during shelter assessments. We examined the impact of shelters omitting food guarding assessments. Nine shelters conducted a two-month baseline period of assessing for food guarding followed by a two-month investigative period during which they omitted the food guarding assessment. Dogs that guarded their food during a standardized assessment were less likely to be adopted, had a longer shelter stay, and were more likely to be euthanized. When the shelters stopped assessing for food guarding, there was no significant difference in the rate of returns of food guarding dogs, even though more dogs were adopted because fewer were identified with food guarding behavior. Additionally, the number of injuries to staff, volunteers, and adopters was low (104 incidents from a total of 14,180 dogs) and did not change when the food guarding assessment was omitted. These results support a recommendation that shelters discontinue the food guarding assessment. PMID:29419746

Overexpression of AtABCG25 enhances the abscisic acid signal in guard cells and improves plant water use efficiency.

Science.gov (United States)

Kuromori, Takashi; Fujita, Miki; Urano, Kaoru; Tanabata, Takanari; Sugimoto, Eriko; Shinozaki, Kazuo

2016-10-01

In addition to improving drought tolerance, improvement of water use efficiency is a major challenge in plant physiology. Due to their trade-off relationships, it is generally considered that achieving stress tolerance is incompatible with maintaining stable growth. Abscisic acid (ABA) is a key phytohormone that regulates the balance between intrinsic growth and environmental responses. Previously, we identified AtABCG25 as a cell-membrane ABA transporter that export ABA from the inside to the outside of cells. AtABCG25-overexpressing plants showed a lower transpiration phenotype without any growth retardation. Here, we dissected this useful trait using precise phenotyping approaches. AtABCG25 overexpression stimulated a local ABA response in guard cells. Furthermore, AtABCG25 overexpression enhanced drought tolerance, probably resulting from maintenance of water contents over the common threshold for survival after drought stress treatment. Finally, we observed enhanced water use efficiency by overexpression of AtABCG25, in addition to drought tolerance. These results were consistent with the function of AtABCG25 as an ABA efflux transporter. This unique trait may be generally useful for improving the water use efficiency and drought tolerance of plants. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

The Impact of Excluding Food Guarding from a Standardized Behavioral Canine Assessment in Animal Shelters

Directory of Open Access Journals (Sweden)

Heather Mohan-Gibbons

2018-02-01

Full Text Available Many shelters euthanize or restrict adoptions for dogs that exhibit food guarding while in the animal shelter. However, previous research showed that only half the dogs exhibiting food guarding during an assessment food guard in the home. So, dogs are often misidentified as future food guarders during shelter assessments. We examined the impact of shelters omitting food guarding assessments. Nine shelters conducted a two-month baseline period of assessing for food guarding followed by a two-month investigative period during which they omitted the food guarding assessment. Dogs that guarded their food during a standardized assessment were less likely to be adopted, had a longer shelter stay, and were more likely to be euthanized. When the shelters stopped assessing for food guarding, there was no significant difference in the rate of returns of food guarding dogs, even though more dogs were adopted because fewer were identified with food guarding behavior. Additionally, the number of injuries to staff, volunteers, and adopters was low (104 incidents from a total of 14,180 dogs and did not change when the food guarding assessment was omitted. These results support a recommendation that shelters discontinue the food guarding assessment.

Penium margaritaceum as a model organism for cell wall analysis of expanding plant cells

DEFF Research Database (Denmark)

Rydahl, Maja Gro; Fangel, Jonatan Ulrik; Mikkelsen, Maria Dalgaard

2015-01-01

organization of the polymeric networks of the cell wall around the protoplast also contributes to the direction of growth, the shape of the cell, and the proper positioning of the cell in a tissue. In essence, plant cell expansion represents the foundation of development. Most studies of plant cell expansion...... have focused primarily upon late divergent multicellular land plants and specialized cell types (e.g., pollen tubes, root hairs). Here, we describe a unicellular green alga, Penium margaritaceum (Penium), which can serve as a valuable model organism for understanding cell expansion and the underlying......The growth of a plant cell encompasses a complex set of subcellular components interacting in a highly coordinated fashion. Ultimately, these activities create specific cell wall structural domains that regulate the prime force of expansion, internally generated turgor pressure. The precise...

30 CFR 56.7013 - Covering or guarding drill holes.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Covering or guarding drill holes. 56.7013 Section 56.7013 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND... Rotary Jet Piercing Drilling § 56.7013 Covering or guarding drill holes. Drill holes large enough to...

30 CFR 57.7013 - Covering or guarding drill holes.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Covering or guarding drill holes. 57.7013 Section 57.7013 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND... and Rotary Jet Piercing Drilling-Surface Only § 57.7013 Covering or guarding drill holes. Drill holes...

Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts

Energy Technology Data Exchange (ETDEWEB)

Steponkus, P.L.

1991-01-01

This project focuses on lesions in the plasma membrane of protoplasts that occur during freezing to temperatures below {minus}5{degrees} which result in changes in the semipermeablity of the plasma membrane. This injury, referred to as loss of osmotic responsiveness, is associated with the formation of large, aparticulate domains in the plasma membrane, aparticulate lamellae subtending the plasma membrane, and lamellar-to-hexagonal{sub II} phase transitions in the plasma membrane and subtending lamellar. The goals of this project are to provide a mechanistic understanding of the mechanism by which freeze-induced dehydration effects the formation of aparticulate domains and lamellar-to-hexagonal{sub II} phase transitions and to determine the mechanisms by which cold acclimation and cryoprotectants preclude or diminish these ultrastructural changes. Our working hypothesis is the formation of aparticulate domains and lamellar-to-hexagon{sub II} phase transitions in the plasma membrane and subtending lamellae are manifestations of hydration-dependent bilayer-bilayer interactions.

Great Lakes Surface Ice Reports from U.S. Coast Guard

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Data consist of ice observations from U.S. Coast Guard vessels operating on the Great Lakes, and from Coast Guard shore stations reported via teletype messages and...

Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion.

Science.gov (United States)

Hatvani, Lóránt; Manczinger, László; Kredics, László; Szekeres, András; Antal, Zsuzsanna; Vágvölgyi, Csaba

2006-01-01

The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T. atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 microg/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 microg/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T. atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.

Coast Guard Proceedings. Volume 70, Number 2, Summer 2013

Science.gov (United States)

2013-01-01

and well-being. Most Inuit are coastal people who rely heavily on resources from the ocean for nutritional and cultural survival. The Inuit are a...defend a soccer ball from a crewmember of the Russian vessel Vorovsky at the Coast Guard gym at Base Kodiak, Alaska. U.S. Coast Guard photo by

Line Heat-Source Guarded Hot Plate

Data.gov (United States)

Federal Laboratory Consortium — Description:The 1-meter guarded hot-plate apparatus measures thermal conductivity of building insulation. This facility provides for absolute measurement of thermal...

Efficient parallel implementations of approximation algorithms for guarding 1.5D terrains

Directory of Open Access Journals (Sweden)

Goran Martinović

2015-03-01

Full Text Available In the 1.5D terrain guarding problem, an x-monotone polygonal line is dened by k vertices and a G set of terrain points, i.e. guards, and a N set of terrain points which guards are to observe (guard. This involves a weighted version of the guarding problem where guards G have weights. The goal is to determine a minimum weight subset of G to cover all the points in N, including a version where points from N have demands. Furthermore, another goal is to determine the smallest subset of G, such that every point in N is observed by the required number of guards. Both problems are NP-hard and have a factor 5 approximation [3, 4]. This paper will show that if the (1+ϵ-approximate solver for the corresponding linear program is a computer, for any ϵ > 0, an extra 1+ϵ factor will appear in the final approximation factor for both problems. A comparison will be carried out the parallel implementation based on GPU and CPU threads with the Gurobi solver, leading to the conclusion that the respective algorithm outperforms the Gurobi solver on large and dense inputs typically by one order of magnitude.

Growth of Avena Coleoptiles and pH Drop of Protoplast Suspensions Induced by Chlorinated Indoleacetic Acids

DEFF Research Database (Denmark)

Engvild, Kjeld Christensen; Doll, Hans; Böttger, M.

1978-01-01

-auxins. Some of the derivatives were compared for their effect on pH decline in stem protoplast suspensions of Helianthus annuus L. and Pisum sativum L. The change of pH occurs without a lag period or with only a very short one. Derivatives which are very active in the Avena straight growth assay cause......Several indoleacetic acids, substituted in the benzene ring, were compared in the Avena straight growth bioassay. 4-Chloroindoleacetic acid, a naturally occurring plant hormone, is one of the strongest hormones in this bioassay. With an optimum at 10-6 mol l-1, it is more active than indoleacetic...... a larger pH decline than indoleacetic acid, while inactive derivatives cause effectively no pH decline....

Surfing for mouth guards: assessing quality of online information.

Science.gov (United States)

Magunacelaya, Macarena B; Glendor, Ulf

2011-10-01

The Internet is an easily accessible and commonly used source of health-related information, but evaluations of the quality of this information within the dental trauma field are still lacking. The aims of this study are (i) to present the most current scientific knowledge regarding mouth guards used in sport activities, (ii) to suggest a scoring system to evaluate the quality of information pertaining to mouth guard protection related to World Wide Web sites and (iii) to employ this scoring system when seeking reliable mouth guard-related websites. First, an Internet search using the keywords 'athletic injuries/prevention and control' and 'mouth protector' or 'mouth guards' in English was performed on PubMed, Cochrane, SvedMed+ and Web of Science to identify scientific knowledge about mouth guards. Second, an Internet search using the keywords 'consumer health information Internet', 'Internet information public health' and 'web usage-seeking behaviour' was performed on PubMed and Web of Science to obtain scientific articles seeking to evaluate the quality of health information on the Web. Based on the articles found in the second search, two scoring systems were selected. Then, an Internet search using the keywords 'mouth protector', 'mouth guards' and 'gum shields' in English was performed on the search engines Google, MSN and Yahoo. The websites selected were evaluated for reliability and accuracy. Of the 223 websites retrieved, 39 were designated valid and evaluated. Nine sites scored 22 or higher. The mean total score of the 39 websites was 14.2. Fourteen websites scored higher than the mean total score, and 25 websites scored less. The highest total score, presented by a Public Institution Web site (Health Canada), was 31 from a maximum possible score of 34, and the lowest score was 0. This study shows that there is a high amount of information about mouth guards on the Internet but that the quality of this information varies. It should be the responsibility

U.S. Coast Guard (USCG) Districts

Data.gov (United States)

Department of Homeland Security — This layer is a polygonal dataset that represents land and maritime boundaries for each representative United States Coast Guard district, which includes district 1,...

49 CFR 1520.15 - SSI disclosed by TSA or the Coast Guard.

Science.gov (United States)

2010-10-01

... 49 Transportation 9 2010-10-01 2010-10-01 false SSI disclosed by TSA or the Coast Guard. 1520.15... PROTECTION OF SENSITIVE SECURITY INFORMATION § 1520.15 SSI disclosed by TSA or the Coast Guard. (a) In... available for public inspection or copying, nor does TSA or the Coast Guard release such records to persons...

30 CFR 57.12023 - Guarding electrical connections and resistor grids.

Science.gov (United States)

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Guarding electrical connections and resistor... NONMETAL MINES Electricity Surface and Underground § 57.12023 Guarding electrical connections and resistor grids. Electrical connections and resistor grids that are difficult or impractical to insulate shall be...

76 FR 17782 - Protection for Whistleblowers in the Coast Guard

Science.gov (United States)

2011-03-31

... ``Keyword'' box, and then clicking ``Search.'' FOR FURTHER INFORMATION CONTACT: If you have questions on..., telephone 202-372-3777, e-mail [email protected] . If you have questions on viewing or submitting... Protection for Whistleblowers in the Coast Guard AGENCY: Coast Guard, DHS. ACTION: Direct final rule...

Metal accumulation in Nitellopsis obtusa cells from the laboratory solution

International Nuclear Information System (INIS)

Marciulioniene, D.; Montvydiene, D.; Ceburnis, D.

2001-01-01

The ability of Nitellopsis obtusa to accumulate heavy metals from the laboratory solution containing ions of Cd2+, Cr6+, Cu2+, Mn2+, Ni2+, Pb2+ and Zn2+ was investigated. Concentrations of heavy metals in the algae cells were determined, and the accumulation coefficient (AC) of heavy metals in the live cells (in the wall and the protoplast), in the dead cells (in the wall), and in the cells which have lost turgor were estimated. It was found that, according to the accumulation coefficient values in the cell wall of N. obtusa, the studied metals followed the order: Cr6+ < Pb2+ < Ni2+ < Cd2+ < Cu2+ < Zn2+ < Mn2+, while according to the accumulation coefficient values in the protoplast, the order was: Pb2+ < Cr6+ < Ni2+ < Zn2+ < Cd2+ < Cu2+ < Mn2+ . It was demonstrated that in both media metals were accumulated very similarly. The difference between AC in the cell walls of the live and dead cells was negligible. The obtained data allowed to conclude that all investigated metals were not only absorbed in the algae cell wall but they were intensively up taken into the cell. Data showed that among all investigated metals Cr6+ was the least absorbed in the cell wall, while Pb was predominantly absorbed in the cell wall, as well as Cd2+ and Cu2+ were more intensively up taken into the cell than other metals It was established that Mn2+ was Intensively adsorbed in the cell wall, and its uptake into the cell was intensive, too. (author)

77 FR 16929 - Safety Zones; Fireworks Displays within the Fifth Coast Guard District

Science.gov (United States)

2012-03-23

...-AA00 Safety Zones; Fireworks Displays within the Fifth Coast Guard District AGENCY: Coast Guard, DHS... for fireworks displays at various locations within the geographic boundary of the Fifth Coast Guard... by fireworks displays. Entry into or movement within these zones during the enforcement periods is...

United States Coast Guard: Officer Corps Military Professional Development Program

Science.gov (United States)

2011-06-10

captain relieved of command after viewing porn on computer,‖ 10 July 2010, http://www.thenewstribune.com/2010/07/ 10/1259108/portland-coast-guard...Coast Guard captain relieved of command after viewing porn on computer,‖ 10 July 2010. http://www.thenewstribune.com/2010/07/ 10/1259108/portland

29 CFR Appendix B to Subpart B of... - Reprint of U.S. Coast Guard Regulations Referenced in Subpart B, for Determination of Coast Guard...

Science.gov (United States)

2010-07-01

... Subpart B, for Determination of Coast Guard Authorized Persons B Appendix B to Subpart B of Part 1915... Enclosed Spaces and Other Dangerous Atmospheres in Shipyard Employment Pt. 1915, Subpt. B, App. B Appendix B to Subpart B of Part 1915—Reprint of U.S. Coast Guard Regulations Referenced in Subpart B, for...

Quantitative Proteomic Analysis of the Response to Zinc, Magnesium, and Calcium Deficiency in Specific Cell Types of Arabidopsis Roots

Directory of Open Access Journals (Sweden)

Yoichiro Fukao

2016-01-01

Full Text Available The proteome profiles of specific cell types have recently been investigated using techniques such as fluorescence activated cell sorting and laser capture microdissection. However, quantitative proteomic analysis of specific cell types has not yet been performed. In this study, to investigate the response of the proteome to zinc, magnesium, and calcium deficiency in specific cell types of Arabidopsis thaliana roots, we performed isobaric tags for relative and absolute quantification (iTRAQ-based quantitative proteomics using GFP-expressing protoplasts collected by fluorescence-activated cell sorting. Protoplasts were collected from the pGL2-GFPer and pMGP-GFPer marker lines for epidermis or inner cell lines (pericycle, endodermis, and cortex, respectively. To increase the number of proteins identified, iTRAQ-labeled peptides were separated into 24 fractions by OFFGFEL electrophoresis prior to high-performance liquid chromatography coupled with mass spectrometry analysis. Overall, 1039 and 737 proteins were identified and quantified in the epidermal and inner cell lines, respectively. Interestingly, the expression of many proteins was decreased in the epidermis by mineral deficiency, although a weaker effect was observed in inner cell lines such as the pericycle, endodermis, and cortex. Here, we report for the first time the quantitative proteomics of specific cell types in Arabidopsis roots.

Triangulating and guarding realistic polygons

NARCIS (Netherlands)

Aloupis, G.; Bose, P.; Dujmovic, V.; Gray, C.M.; Langerman, S.; Speckmann, B.

2014-01-01

We propose a new model of realistic input: k-guardable objects. An object is k-guardable if its boundary can be seen by k guards. We show that k-guardable polygons generalize two previously identified classes of realistic input. Following this, we give two simple algorithms for triangulating

29 CFR 1915.73 - Guarding of deck openings and edges.

Science.gov (United States)

2010-07-01

... 29 Labor 7 2010-07-01 2010-07-01 false Guarding of deck openings and edges. 1915.73 Section 1915.73 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH ADMINISTRATION..., Ladders and Other Working Surfaces § 1915.73 Guarding of deck openings and edges. (a) The provisions of...

Penium margaritaceum as a model organism for cell wall analysis of expanding plant cells.

Science.gov (United States)

Rydahl, Maja G; Fangel, Jonatan U; Mikkelsen, Maria Dalgaard; Johansen, I Elisabeth; Andreas, Amanda; Harholt, Jesper; Ulvskov, Peter; Jørgensen, Bodil; Domozych, David S; Willats, William G T

2015-01-01

The growth of a plant cell encompasses a complex set of subcellular components interacting in a highly coordinated fashion. Ultimately, these activities create specific cell wall structural domains that regulate the prime force of expansion, internally generated turgor pressure. The precise organization of the polymeric networks of the cell wall around the protoplast also contributes to the direction of growth, the shape of the cell, and the proper positioning of the cell in a tissue. In essence, plant cell expansion represents the foundation of development. Most studies of plant cell expansion have focused primarily upon late divergent multicellular land plants and specialized cell types (e.g., pollen tubes, root hairs). Here, we describe a unicellular green alga, Penium margaritaceum (Penium), which can serve as a valuable model organism for understanding cell expansion and the underlying mechanics of the cell wall in a single plant cell.

Work-related violence against security guards--who is most at risk?

Science.gov (United States)

Leino, Tuula; Selin, Risto; Summala, Heikki; Virtanen, Marianna

2011-01-01

Studies on violence in the work of security guards are largely lacking. This study is unique in that it focuses on security guards (n=1,010) in Finland, and assesses the different forms, prevalence, and risk factors of the work-related violence they often face. Information to a survey instrument was obtained by first interviewing 30 volunteers. Then we made a cross-sectional mailed survey that was sent to a randomized group of 2,000 security guards. The response rate was 52. We found the prevalence of verbal aggression, threats of assault, and physical acts against security guards at least once a month to be 39%, 19%, and 15% respectively. As regards risk factors and who is most at risk, our results show that male gender, young age, low work experience, late working hours, and time pressure were associated with all three forms of work-related violence. Unlike other forms of violence, verbal aggression was highly prevalent outside the metropolitan area and directed towards both more and less experienced security guards. In prevention policies for violence, it is important to identify high-risk groups such as those who have less work experience.

Triangulating and guarding realistic polygons

NARCIS (Netherlands)

Aloupis, G.; Bose, P.; Dujmovic, V.; Gray, C.M.; Langerman, S.; Speckmann, B.

2008-01-01

We propose a new model of realistic input: k-guardable objects. An object is k-guardable if its boundary can be seen by k guards in the interior of the object. In this abstract, we describe a simple algorithm for triangulating k-guardable polygons. Our algorithm, which is easily implementable, takes

Relational Reasoning for Markov Chains in a Probabilistic Guarded Lambda Calculus

DEFF Research Database (Denmark)

Aguirre, Alejandro; Barthe, Gilles; Birkedal, Lars

2018-01-01

We extend the simply-typed guarded $\\lambda$-calculus with discrete probabilities and endow it with a program logic for reasoning about relational properties of guarded probabilistic computations. This provides a framework for programming and reasoning about infinite stochastic processes like...

The continuous 1.5D terrain guarding problem: Discretization, optimal solutions, and PTAS

Directory of Open Access Journals (Sweden)

Stephan Friedrichs

2016-05-01

Full Text Available In the NP-hard continuous 1.5D Terrain Guarding Problem (TGP we are given an $x$-monotone chain of line segments in $R^2$ (the terrain $T$, and ask for the minimum number of guards (located anywhere on $T$ required to guard all of $T$. We construct guard candidate and witness sets $G, W \\subset T$ of polynomial size such that any feasible (optimal guard cover $G^* \\subseteq G$ for $W$ is also feasible (optimal for the continuous TGP. This discretization allows us to: (1 settle NP-completeness for the continuous TGP; (2 provide a Polynomial Time Approximation Scheme (PTAS for the continuous TGP using the PTAS for the discrete TGP by Gibson et al.; (3 formulate the continuous TGP as an Integer Linear Program (IP. Furthermore, we propose several filtering techniques reducing the size of our discretization, allowing us to devise an efficient IP-based algorithm that reliably provides optimal guard placements for terrains with up to $10^6$ vertices within minutes on a standard desktop computer.

Implementing Strategy in a Budget: A Model of the Coast Guard Reserve

OpenAIRE

Bromund, Carl Douglas

1990-01-01

Approved for public release; distribution is unlimited. This thesis discusses the managment strategy of the Coast Guard Reserve; it examines the formulation and implmentation of strateqy. A model to develop and implement strategy is proposed, which defines the role of the budget in this strategic management process. The recent strategy of the Coast Guard Reserve is analyzed using this model.. This analysis seems to indicate that the Coast Guard Reserve had no explicit strate...

Automated intracranial pressure-controlled cerebrospinal fluid external drainage with LiquoGuard.

Science.gov (United States)

Linsler, Stefan; Schmidtke, Mareike; Steudel, Wolf Ingo; Kiefer, Michael; Oertel, Joachim

2013-08-01

LiquoGuard is a new device for intracranial pressure (ICP)-controlled drainage of cerebrospinal fluid (CSF). This present study evaluates the accuracy of ICP measurement via the LiquoGuard device in comparison with Spiegelberg. Thus, we compared data ascertained from simultaneous measurement of ICP using tip-transducer and tip-sensor devices. A total of 1,764 monitoring hours in 15 patients (range, 52-219 h) were analysed. All patients received an intraventricular Spiegelberg III probe with the drainage catheter connected to the LiquoGuard system. ICP reading of both devices was performed on an hourly basis. Statistical analysis was done by applying Pearson correlation and Wilcoxon-matched pair test (p drainage. However, LiquoGuard tends to provide misleading results in slit ventricles. Thus, before these drawbacks are further analysed, the authors recommend additional ICP measurement with internal tip-sensor devices to avoid dangerous erroneous interpretation of ICP data.

Learning Without Boundaries: A NASA - National Guard Bureau Distance Learning Partnership

Science.gov (United States)

Anderson, Susan H.; Chilelli, Christopher J.; Picard, Stephan

2003-01-01

With a variety of high-quality live interactive educational programs originating at the Johnson Space Center in Houston, Texas and other space and research centers, the US space agency NASA (National Aeronautics and Space Administration) has a proud track record of connecting with students throughout the world and stimulating their creativity and collaborative skills by teaching them underlying scientific and technological underpinnings of space exploration. However, NASA desires to expand its outreach capability for this type of interactive instruction. In early 2002, NASA and the National Guard Bureau -- using the Guard's nationwide system of state-ofthe-art classrooms and high bandwidth network -- began a collaboration to extend the reach of NASA content and educational programs to more of America's young people. Already, hundreds of elementary, middle, and high school students have visited Guard e-Learning facilities and participated in interactive NASA learning events. Topics have included experimental flight, satellite imagery-interpretation, and Mars exploration. Through this partnership, NASA and the National Guard are enabling local school systems throughout the United States (and, increasingly, the world) to use the excitement of space flight to encourage their students to become passionate about the possibility of one day serving as scientists, mathematicians, technologists, and engineers. At the 54th International Astronautical Conference MAJ Stephan Picard, the guiding visionary behind the Guard's partnership with NASA, and Chris Chilelli, an educator and senior instructional designer at NASA, will share with attendees background on NASA's educational products and the National Guard's distributed learning network; will discuss the unique opportunity this partnership already has provided students and teachers throughout the United States; will offer insights into the formation by government entities of e-Learning partnerships with one another; and will

5 CFR 831.306 - Service as a National Guard technician before January 1, 1969.

Science.gov (United States)

2010-01-01

... 5 Administrative Personnel 2 2010-01-01 2010-01-01 false Service as a National Guard technician... National Guard technician before January 1, 1969. (a) Definitions. In this section—(1) Service as a National Guard technician is service performed under section 709 of title 32, United States Code (or under...

Role of guard rings in improving the performance of silicon detectors

Indian Academy of Sciences (India)

number of free charge carriers thus produced is proportional to the energy deposited. 259 .... grounding the innermost guard ring of a strip detector of geometry 60 × 60 mm2. ... The guard ring was kept floating with no electrical connection .... ment of maintaining high-quality semiconductor properties during high temperature.

Perceptions of Individual and Family Functioning Among Deployed Female National Guard Members.

Science.gov (United States)

Kelly, Patricia J; Cheng, An-Lin; Berkel, LaVerne A; Nilsson, Johanna

2016-08-01

Females currently make up 15% of U.S. military service members. Minimal attention has been paid to families of female National Guard members who have been deployed and their subsequent reintegration challenges. This cross-sectional Internet-based survey of female members of four National Guard units compared those who were and were not deployed. Instruments, guided by the variables of the Family Resilience Model, measured individual, family, and deployment-related factors. Bivariate analysis and ordinal logistic regression were done to assess differences between the groups. Of the 239 National Guard members surveyed, deployed women (n = 164) had significantly higher levels of posttraumatic stress disorder (PTSD; p family functioning were higher among deployed when compared with never deployed women. Results indicate community interventions that focus on strengthening coping skills of female Guard members would be useful for this population. © The Author(s) 2016.

Joint FAM/Line Management Assessment Report on LLNL Machine Guarding Safety Program

Energy Technology Data Exchange (ETDEWEB)

Armstrong, J. J. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

2016-07-19

The LLNL Safety Program for Machine Guarding is implemented to comply with requirements in the ES&H Manual Document 11.2, "Hazards-General and Miscellaneous," Section 13 Machine Guarding (Rev 18, issued Dec. 15, 2015). The primary goal of this LLNL Safety Program is to ensure that LLNL operations involving machine guarding are managed so that workers, equipment and government property are adequately protected. This means that all such operations are planned and approved using the Integrated Safety Management System to provide the most cost effective and safest means available to support the LLNL mission.

Early Intervention to Reduce Alcohol Misuse and Abuse in the Ohio Army National Guard

Science.gov (United States)

2017-09-01

PTSD, depression , 7 suicidality , lowered resilience and work performance.  Studies consistently find that substantial number of Guard members misuse...including PTSD, depression , suicidality , lowered resilience and work performance.  Studies consistently find that substantial number of Guard members misuse...Guard members that contributes to and complicates other problems including PTSD, depression , suicidality , lowered resilience and work performance

29 CFR 1910.23 - Guarding floor and wall openings and holes.

Science.gov (United States)

2010-07-01

... 29 Labor 5 2010-07-01 2010-07-01 false Guarding floor and wall openings and holes. 1910.23 Section... floor and wall openings and holes. (a) Protection for floor openings. (1) Every stairway floor opening... opening and hole shall be guarded by a standard skylight screen or a fixed standard railing on all exposed...

In vitro mutation breeding for salinity tolerance in Citrus

International Nuclear Information System (INIS)

Deng Zhanao; Zhang Wencai; Wan Shuyan

1989-01-01

Full text: Mutation breeding began in the 1970's, concentrating on early ripening and high quality spontaneous mutants, subsequently on the induction of seedless mutants by gamma radiation and other mutagens. Extremely early cultivars 'Guoqing 1 to 5' have been obtained from bud mutations in satsuma mandarin and are grown commercially. Other desirable mutants have been found in progenies from gamma ray treated budwood. Recently we introduced plant tissue and cell culture techniques hoping to avoid chimerism and diplontic selection and to obtain solid mutants in a shorter time, resistant to salt, herbicides, low temperature and diseases. I. Methods for cell and protoplast culture. Ovules from fruitlets 1-6 weeks after flowering, cultured in MT supplemented with IAA 0.1 mg/I and KT 1.0 mg/l, gradually produce callus from nucelli. After several passages, habituated callus lines are obtained, which grow very well in MT medium without any growth substances. These callus lines, regardless of the species and cultivars are white, friable, composed of cell clumps of various size and highly embryogenic. When transferred to MT medium containing 2% glycerol and caseinhydrolysat 400 mg/l, they become green and form numerous green globular embryoids within one month. Most of the embryoids derive from single cells. The embryoids could develop roots and shoots forming large numbers of plantlets in a fresh medium. Habituated callus lines have been established in 5 cultivars. Their high embryogenic capacity of single cell origin provides good material for mutation induction. For obtaining protoplasts, the habituated calluses were transferred into liquid MT medium and cultured for two passages on a shaker, then they were macerated with enzyme solution containing 0.3%-0.5% pectinase, 0.3%-0.5% cellulase, MT macroelements and 0.7 M mannitol, at 5.7 pH. After incubation at 25 deg. C for 12-16 hrs., a large number of viable protoplasts could be collected. They were resuspended to a

ARMY NATIONAL GUARD: Enhanced Brigade Readiness Improved but Personnel and Workload Are Problems

National Research Council Canada - National Science Library

2000-01-01

.... Reserve units such as the National Guard's 15 Enhanced Separate Brigades, the Guard's highest priority combat units, provide fighting forces at about 2530 percent of the cost of active units due...

Relating Operational Art to the National Guard State Partnership Program

Science.gov (United States)

2014-05-15

Program Coordinator GEF Global Employment of the Force DSCA Defense Support of Civil Authorities BCA Budget Control Act NATO North Atlantic Treaty...between Florida and the U.S. Virgin Islands are referred to as the Regional Security System (RSS). Florida and the U.S. Virgin Islands partner with... Atlantic Treaty Organization (NATO) in 2004 due, in part, to the mentorship from the Maryland National Guard.54 Not only did the Maryland National Guard

Preliminary assessment report for National Guard Training Center, Georgia Army National Guard, Fort Stewart, Georgia

International Nuclear Information System (INIS)

1993-07-01

This report presents the results of the preliminary assessment (PA) conducted by Argonne National Laboratory at the Georgia Army National Guard (GAARNG) facility near Hinesville, Georgia, known as the National Guard Training Center (NGTC). Preliminary assessments of federal facilities are being conducted to compile the information necessary for completing preremedial activities and to provide a priority basis for completing corrective actions (where necessary) in response to releases of hazardous substances. The principal objective of the PA is to characterize the site accurately and determine the need for further action by examining previous site activities, types and quantities of hazardous substances utilized, and potential pathways by which contamination could affect public health and the environment. The scope of this assessment is limited to the facilities and past activities contained within the NGTC. Preliminary assessment site score sheet information is also provided for the NGTC. However, this assessment report is intended to be read in conjunction with a previous IRP assessment of Fort Stewart completed in 1992 (USATHAMA 1992) and to provide comprehensive information on the NGTC area for incorporation with information contained in that previous assessment for the entirety of Fort Stewart

48 CFR 3009.171-3 - Determination of eligibility for award of FPS guard service contracts.

Science.gov (United States)

2010-10-01

... eligibility for award of FPS guard service contracts. 3009.171-3 Section 3009.171-3 Federal Acquisition... eligibility for award of FPS guard service contracts. (a) Contracting officers shall make a determination of eligibility for award of FPS guard service contracts upon identification of the apparent successful offeror as...

AKTIVITAS ENZIM KOMERSIAL, EKSTRAK KASAR ENZIM DARI VISCERA KEONG MAS (Pila polita, ABALON (Haliotis asinina, DAN BEKICOT (Achatina fulica UNTUK LISIS JARINGAN RUMPUT LAUT Kappaphycus alvarezii PADA KULTUR PROTOPLAS

Directory of Open Access Journals (Sweden)

Sri Redjeki Hesti Mulyaningrum

2008-12-01

(Haliotis asinina 0.014 unit/mL activity. Optimum ratio of commercial enzyme was 2:1, it resulted protoplast up to 1.26 x 108 cell/mL, then ratio of 1:2 resulted protoplast up to 1.22 x 108 cell/mL, ratio of 1:1 resulted protoplast up to 8.36 x 107 cell/mL, ratio of 0:1 resulted protoplast up to 6.33 x 107 cell/mL and ratio of 1:0 resulted protoplast up to 9.55 x 106 cell/mL. The highest density of protoplast gained by seaweed from Takalar reached 3.7 x 108 cell/mL.

Extending the fungal host range of a partitivirus and a mycoreovirus from Rosellinia necatrix by inoculation of protoplasts with virus particles.

Science.gov (United States)

Kanematsu, Satoko; Sasaki, Atsuko; Onoue, Mari; Oikawa, Yuri; Ito, Tsutae

2010-09-01

The potential host range of mycoviruses is poorly understood because of the lack of suitable inoculation methods. Recently, successful transfection has been reported for somatically incompatible fungal isolates with purified virus particles of two mycoviruses, the partitivirus RnPV1-W8 (RnPV1) and the mycoreovirus RnMyRV3/W370 (MyRV3), from the white root rot fungus Rosellinia necatrix (class Sordariomycetes, subclass Xylariomycetidae). These studies examined and revealed the effect of the mycoviruses on growth and pathogenicity of R. necatrix. Here, we extended the experimental host range of these two mycoviruses using a transfection approach. Protoplasts of other phytopathogenic Sordariomycetous fungi-Diaporthe sp., Cryphonectria parasitica, Valsa ceratosperma (Sordariomycetidae), and Glomerella cingulata (Hypocreomycetidae)-were inoculated with RnPV1 and MyRV3 viral particles. The presence of double-stranded RNA viral genomes in regenerated mycelia of Diaporthe sp., C. parasitica, and V. ceratosperma confirmed both types of viral infections in these three novel host species. An established RnPV1 infection was confirmed in G. cingulata but MyRV3 did not infect this host. Horizontal transmission of both viruses from newly infected strains to virus-free, wild-type strains through hyphal anastomosis was readily achieved by dual culture; however, vertical transmission through conidia was rarely observed. The virulence of Diaporthe sp., C. parasitica, and V. ceratosperma strains harboring MyRV3 was reduced compared with their virus-free counterpart. In summary, our protoplast inoculation method extended the experimental host range of RnPV1-W8 and MyRV3 within the class Sordariomycetes and revealed that MyRV3 confers hypovirulence to the new hosts, as it does to R. necatrix.

76 FR 7131 - Safety Zones; Eleventh Coast Guard District Annual Fireworks Events

Science.gov (United States)

2011-02-09

...-AA00 Safety Zones; Eleventh Coast Guard District Annual Fireworks Events AGENCY: Coast Guard, DHS... permanent safety zones to ensure public safety during annual firework displays at various locations in the... events, delete events that are no longer occurring, add new unlisted annual fireworks events to the...

Construction and evaluation of an exopolysaccharide-producing engineered bacterial strain by protoplast fusion for microbial enhanced oil recovery.

Science.gov (United States)

Sun, Shanshan; Luo, Yijing; Cao, Siyuan; Li, Wenhong; Zhang, Zhongzhi; Jiang, Lingxi; Dong, Hanping; Yu, Li; Wu, Wei-Min

2013-09-01

Enterobacter cloacae strain JD, which produces water-insoluble biopolymers at optimal temperature of 30°C, and a thermophilic Geobacillus strain were used to construct an engineered strain for exopolysaccharide production at high temperatures by protoplast fusion. The obtained fusant strain ZR3 produced exopolysaccharides at up to 45°C with optimal growth temperature at 35°C. The fusant produced exopolysaccharides of approximately 7.5 g/L or more at pH between 7.0 and 9.0. The feasibility of the enhancement of crude oil recovery with the fusant was tested in a sand-packed column at 40°C. The results demonstrated that bioaugmentation of the fusant was promising approach for MEOR. Mass growth of the fusant was confirmed in fermentor tests. Copyright © 2013 Elsevier Ltd. All rights reserved.

75 FR 53195 - Security Zone; U.S. Coast Guard BSU Seattle, Pier 36, Seattle, WA

Science.gov (United States)

2010-08-31

... its effects on them and participate in the rulemaking process. Small businesses may send comments on... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 [Docket No. USCG-2010-0021] RIN 1625-AA87 Security Zone; U.S. Coast Guard BSU Seattle, Pier 36, Seattle, WA AGENCY: Coast Guard, DHS. ACTION...

33 CFR 165.506 - Safety Zones; Fifth Coast Guard District Fireworks Displays.

Science.gov (United States)

2010-07-01

... District Fireworks Displays. 165.506 Section 165.506 Navigation and Navigable Waters COAST GUARD... § 165.506 Safety Zones; Fifth Coast Guard District Fireworks Displays. (a) Regulations. The following regulations apply to the fireworks safety zones listed in the Table to § 165.506. (1) The general regulations...

33 CFR 23.15 - Coast Guard ensign.

Science.gov (United States)

2010-07-01

... distinctive emblem of the Coast Guard in blue and white is placed with its center on a line with the lower..., the emblem covering a horizontal space of three stripes. [CGFR 57-35, 22 FR 6765, Aug. 22, 1957] ...

Modeling and Design Guidelines for P⁺ Guard Rings in Lightly Doped CMOS Substrates

DEFF Research Database (Denmark)

Shen, Ming; Mikkelsen, Jan H.; Zhang, Ke

2013-01-01

of ${rm P}^{+}$ guard rings in terms of S-parameters, which is useful for substrate noise mitigation in mixed-signal system-on-chips. Validation of the model has been done by both electromagnetic simulation and experimental results from guard rings implemented using a standard 0.18-$mu{rm m}$ CMOS process....... In addition, design guidelines have been drawn for minimizing the guard ring size while maintaining the noise suppression performance....

Somatic hybridization in Citrus: navel orange (C. sinensis Osb.) and grapefruit (C. paradisi Macf.).

Science.gov (United States)

Ohgawara, T; Kobayashi, S; Ishii, S; Yoshinaga, K; Oiyama, I

1989-11-01

Protoplasts of navel orange, isolated from embryogenic nucellar cell suspension culture, were fused with protoplasts of grapefruit isolated from leaf tissue. The fusion products were cultured in the hormone-free medium containing 0.6 M sucrose. Under the culture conditions, somatic embryogenesis of navel orange protoplasts was suppressed, while cell division of grapefruit mesophyll protoplasts was not induced. Six embryoids were obtained and three lines regenerated to complete plants through embryogenesis. Two of the regenerated lines exhibited intermediate morphological characteristics of the parents in the leaf shape. Chromosome counts showed that these regenerated plants had expected 36 chromosomes (2n=2x=18 for each parent). The rDNA analysis using biotin-labeled rRNA probes confirmed the presence of genomes from both parents in these plants. This somatic hybridization system would be useful for the practical Citrus breeding.

78 FR 8027 - Safety Zone, Coast Guard Exercise Area, Hood Canal, Washington

Science.gov (United States)

2013-02-05

... 5, 2012. The Coast Guard received zero comments submitted via regulations.gov and received zero... zero comments from the Small Business Administration on this rule. The Coast Guard certifies under 5 U... more in any one year. Though this rule will not result in such an expenditure, we do discuss the...

Inhibition of cadmium ion uptake in rice (Oryza sativa) cells by a wall-bound form of silicon.

Science.gov (United States)

Liu, Jian; Ma, Jie; He, Congwu; Li, Xiuli; Zhang, Wenjun; Xu, Fangsen; Lin, Yongjun; Wang, Lijun

2013-11-01

The stresses acting on plants that are alleviated by silicon (Si) range from biotic to abiotic stresses, such as heavy metal toxicity. However, the mechanism of stress alleviation by Si at the single-cell level is poorly understood. We cultivated suspended rice (Oryza sativa) cells and protoplasts and investigated them using a combination of plant nutritional and physical techniques including inductively coupled plasma mass spectrometry (ICP-MS), the scanning ion-selective electrode technique (SIET) and X-ray photoelectron spectroscopy (XPS). We found that most Si accumulated in the cell walls in a wall-bound organosilicon compound. Total cadmium (Cd) concentrations in protoplasts from Si-accumulating (+Si) cells were significantly reduced at moderate concentrations of Cd in the culture medium compared with those from Si-limiting (-Si) cells. In situ measurement of cellular fluxes of the cadmium ion (Cd(2+) ) in suspension cells and root cells of rice exposed to Cd(2+) and/or Si treatments showed that +Si cells significantly inhibited the net Cd(2+) influx, compared with that in -Si cells. Furthermore, a net negative charge (charge density) within the +Si cell walls could be neutralized by an increase in the Cd(2+) concentration in the measuring solution. A mechanism of co-deposition of Si and Cd in the cell walls via a [Si-wall matrix]Cd co-complexation may explain the inhibition of Cd ion uptake, and may offer a plausible explanation for the in vivo detoxification of Cd in rice. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

46 CFR 159.007-7 - Application for acceptance for production inspections and tests: Coast Guard action.

Science.gov (United States)

2010-10-01

... and tests: Coast Guard action. 159.007-7 Section 159.007-7 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL APPROVAL....007-7 Application for acceptance for production inspections and tests: Coast Guard action. (a) From...

Meeting the Organizational Needs of the National Guard as Part of Transformation to the Future Force

National Research Council Canada - National Science Library

Wong, Martha

2005-01-01

.... They must build and maintain U.S. defenses beyond challenge. As part of the total military force and to comply with the National Security Strategy, the National Guard also must transform from the Guard of the past to the Guard of the future...

Electrical crosstalk in front-illuminated photodiode array with different guard ring designs for medical CT applications

International Nuclear Information System (INIS)

Ji Fan; Juntunen, Mikko; Hietanen, Iiro

2009-01-01

This paper presents electrical crosstalk studies on front-illuminated photodiode arrays for medical computed tomography (CT) applications. Crosstalk is an important factor to the system noise and image quality. The electrical crosstalk depends on silicon substrate properties and photodiode structures. The photodiode samples employed in this paper are planar processed on high-resistivity n-type silicon substrate, resulting in a p+/n-/n+ diode structure. Two types of guard ring structures are designed and applied to the same geometry of two-dimensional photodiode arrays. One structure is an n guard ring in the gap area between pixels, and the other structure is an additional p+ guard ring around each pixel together with the n guard ring. A 10 μm light spot with wavelength of 525 nm is used to scan across the surface of the photodiode array in the electrical crosstalk measurements. The electrical currents of two neighbor pixels are measured and the results are compared between two guard ring designs. The design with the p+ guard ring structure gives better electrical crosstalk suppression. Moreover, the measurement results show much smaller influence on surrounding pixels with the p+ guard ring structure in the case of disconnected pixel. Besides the electrical crosstalk, the light sensitivity within the gap area is also discussed between two guard ring designs.

On semantics and applications of guarded recursion

DEFF Research Database (Denmark)

Bizjak, Aleš

2016-01-01

denotational model and a logic for reasoning about program equivalence. In the last three chapters we study syntax and semantics of a dependent type theory with a family of later modalities indexed by the set of clocks, and clock quantifiers. In the fourth and fifth chapters we provide two model constructions......In this dissertation we study applications and semantics of guarded recursion, which is a method for ensuring that self-referential descriptions of objects define a unique object. The first two chapters are devoted to applications. We use guarded recursion, first in the form of explicit step......-indexing and then in the form of the internal language of particular sheaf topos, to construct logical relations for reasoning about contextual approximation of probabilistic and nondeterministic programs. These logical relations are sound and complete and useful for showing a range of example equivalences. In the third...

Denotational semantics in Synthetic Guarded Domain Theory

DEFF Research Database (Denmark)

Paviotti, Marco

In functional programming, features such as recursion, recursive types and general references are central. To define semantics of this kind of languages one needs to come up with certain definitions which may be non-trivial to show well-defined. This is because they are circular. Domain theory has...... been used to solve this kind of problems for specific languages, unfortunately, this technique does not scale for more featureful languages, which prevented it from being widely used. Step-indexing is a more general technique that has been used to break circularity of definitions. The idea is to tweak...... the definition by adding a well- founded structure that gives a handle for recursion. Guarded dependent Type Theory (gDTT) is a type theory which implements step-indexing via a unary modality used to guard recursive definitions. Every circular definition is well-defined as long as the recursive variable...

Findings From the National Machine Guarding Program?A Small Business Intervention

OpenAIRE

Parker, David L.; Yamin, Samuel C.; Xi, Min; Brosseau, Lisa M.; Gordon, Robert; Most, Ivan G.; Stanley, Rodney

2016-01-01

Objectives: The purpose of this nationwide intervention was to improve machine safety in small metal fabrication businesses (3 to 150 employees). The failure to implement machine safety programs related to guarding and lockout/tagout (LOTO) are frequent causes of Occupational Safety and Health Administration (OSHA) citations and may result in serious traumatic injury. Methods: Insurance safety consultants conducted a standardized evaluation of machine guarding, safety programs, and LOTO. Busi...

Mate guarding in the Seychelles warbler is energetically costly and adjusted to paternity risk.

Science.gov (United States)

Komdeur, J

2001-10-22

Males may increase their fitness through extra-pair copulations (copulations outside the pair bond) that result in extra-pair fertilizations, but also risk lost paternity when they leave their own mate unguarded. The fitness costs of cuckoldry for Seychelles warblers (Acrocephalus sechellensis) are considerable because warblers have a single-egg clutch and, given the short breeding season, no time for a successful replacement clutch. Neighbouring males are the primary threat to a male's genetic paternity. Males minimize their loss of paternity by guarding their mates to prevent them from having extra-pair copulations during their fertile period. Here, I provide experimental evidence that mate-guarding behaviour is energetically costly and that the expression of this trade-off is adjusted to paternity risk (local male density). Free-living males that were induced to reduce mate guarding spent significantly more time foraging and gained significantly better body condition than control males. The larger the reduction in mate guarding, the more pronounced was the increase in foraging and body condition (accounting for food availability). An experimental increase in paternity risk resulted in an increase in mate-guarding intensity and a decrease in foraging and body condition, and vice versa. This is examined using both cross-sectional and longitudinal data. This study on the Seychelles warbler offers experimental evidence that mate guarding is energetically costly and adjusted to paternity risk.

Factors influencing resident's decision to reside in gated and guarded community

Science.gov (United States)

Shamsudin, Zarina; Shamsudin, Shafiza; Zainal, Rozlin

2017-10-01

Gated communities are residential areas developed with restricted access with strictly controlled entrances and surrounded by a close perimeter of wall or fences. Developers, conscious of the need to fulfill the requirement of living in modern and sophisticated lifestyle and gated properties become the trend and mushroomed over the past decade. Nowadays, it is obvious that gated and guarded communities become almost a dominant feature of Malaysia housing development projects. The focus of this paper is to identify the factors contribute resident's decision to reside in gated and guarded community and to study social interaction among gated communities' residents. 150 questionnaires were distributed to the residents of selected gated and guarded community area in order to achieve the objectives and analyzed by using Statistical Package for Social Science (SPSS) and descriptive analysis. The result was tabulated and presented in charts and graphs for a clear and better understanding. The five main factors contribute to resident decision to reside in gated communities were identified and ranked; there are privacy, security, location, lifestyle and prestige. Besides, the residents are feeling neutral towards the facilities and services provided in their gated and guarded residential area. A comprehensive improvement towards the facilities and services is needed to reach higher satisfaction from the residents.

From Darkness to Light: Posttraumatic Growth among Recently Deployed Army National Guard Soldiers

Science.gov (United States)

2013-02-14

Stress and Associated Functioning of Army National Guard Following Exposure to Iraq Warzone Trauma,” Traumatology 14 (2008): 51. 9. Army National...Associated Functioning of Army National Guard Following Exposure to Iraq Warzone Trauma.” Traumatology , 14 (2008): 51–56. Ouimette, Paige, Dawne Vogt

EPA Region 1 Coast Guard Jurisdictional Boundary - Polygons

Data.gov (United States)

U.S. Environmental Protection Agency — Jurisdictional boundary between EPA and Coast Guard for EPA Region I. Created from 1:100000 USGS DLGs with greater detail drawn from 1:24000 commercial street data...

EPA Region 1 Coast Guard Jurisdictional Boundary - Arcs

Data.gov (United States)

U.S. Environmental Protection Agency — Jurisdictional boundary between EPA and Coast Guard for EPA Region I. Created from 1:100000 USGS DLGs with greater detail drawn from 1:24000 commercial street data...

33 CFR 165.776 - Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico 165.776 Section 165.776 Navigation and Navigable Waters COAST... Guard District § 165.776 Security Zone; Coast Guard Base San Juan, San Juan Harbor, Puerto Rico (a...

76 FR 66239 - Safety Zones; Fireworks Displays Within the Fifth Coast Guard District

Science.gov (United States)

2011-10-26

...-AA00 Safety Zones; Fireworks Displays Within the Fifth Coast Guard District AGENCY: Coast Guard, DHS... safety zones established for fireworks displays at various locations within the geographic boundary of... maritime public from the hazards posed by fireworks displays. Entry into or movement within these proposed...

Differential compartmentation of sucrose and gentianose in the cytosol and vacuoles of storage root protoplasts from Gentiana Lutea L.

Science.gov (United States)

Keller, F; Wiemken, A

1982-12-01

The storage roots of perennial Gentiana lutea L.plants contain several sugars. The predominant carbohydrate reserve is gentianose (β-D-glucopyranosyl-(1 → 6)-α-D-glucopyranosyl-(1 ↔ 2)-β-D-fructofuranoside). Vacuoles were isolated from root protoplasts and purified through a betaine density gradient. The yield was about 75%. Gentianose and gentiobiose were localized to 100% in the vacuoles, fructose and glucose to about 80%, and sucrose to only about 50%. Taking the volumes of the vacuolar and extravacuolar (cytosolic) compartments into account it is inferred that gentianose is located exclusively in the vacuoles, whilst sucrose is much more concentrated in the cytosol where it may play a role as a cryoprotectant. The concentration of fructose and glucose appeared to be similar on both sides of the tonoplast.

76 FR 28795 - Privacy Act of 1974; Department of Homeland Security United States Coast Guard-024 Auxiliary...

Science.gov (United States)

2011-05-18

... 1974; Department of Homeland Security United States Coast Guard-024 Auxiliary Database System of... Security/United States Coast Guard-024 Auxiliary Database (AUXDATA) System of Records.'' This system of...: United States Coast Guard Auxiliary Database (AUXDATA). Security classification: Unclassified. System...

Response capabilities of the National Guard: a focus on domestic disaster medical response.

Science.gov (United States)

Bochicchio, Daniel

2010-01-01

The National Guard has a 373-year history of responding to the nation's call to duty for service both at home and abroad (The National Guard Bureau Web site: Available at http://www.ngb.army.mil/default. aspx.). The National Guard (NG) is a constitutionally unique organization (United States Constitution, US Government Printing Office Web site: Available at http://www.gpoaccess.gov/constitution/index.html.). Today's Guard conducts domestic disaster response and civilian assistance missions on a daily basis. Yet, the NG's role, mission, and capabilities are not well-known or understood. The National Response Framework (NRF) places significant responsibility on the local and state disaster planners (Department of Homeland Security: National Response Framework. US Department of Homeland Security, Washington, DC, January 2008). The public health professionals are an integral component of the disaster planning community. It is critical that the public health community be knowledgeable of types and capabilities of all the response assets at their disposal.

78 FR 55214 - Annual Marine Events in the Eighth Coast Guard District, Sabine River; Orange, TX

Science.gov (United States)

2013-09-10

... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 100 [Docket No. USCG-2013-0723] Annual Marine Events in the Eighth Coast Guard District, Sabine River; Orange, TX AGENCY: Coast Guard, DHS... Neches River in Orange, TX from 3 p.m. on September 20, 2013, through 6 p.m. on September 22, 2013. This...

77 FR 47519 - Annual Marine Events in the Eighth Coast Guard District, Sabine River; Orange, TX

Science.gov (United States)

2012-08-09

... DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 100 [Docket No. USCG-2012-0656] Annual Marine Events in the Eighth Coast Guard District, Sabine River; Orange, TX AGENCY: Coast Guard, DHS... Regulations for the S.P.O.R.T. Power Boat Neches River in Orange, TX from 3 p.m. on September 21, 2012...

46 CFR 50.10-30 - Coast Guard number.

Science.gov (United States)

2010-10-01

... boilers and pressure vessels. (c) The Coast Guard number is comprised of the following: (1) Three capital... St. Louis. STB Sturgeon Bay. TAM Tampa. TOL Toledo. VAL Valdez. WNC Wilmington, NC. [CGFR 68-82, 33...

CRISPR/Cas9-based knockouts reveal that CpRLP1 is a negative regulator of the sex pheromone PR-IP in the Closterium peracerosum-strigosum-littorale complex.

Science.gov (United States)

Kanda, Naho; Ichikawa, Machiko; Ono, Ayaka; Toyoda, Atsushi; Fujiyama, Asao; Abe, Jun; Tsuchikane, Yuki; Nishiyama, Tomoaki; Sekimoto, Hiroyuki

2017-12-19

Heterothallic strains of the Closterium peracerosum-strigosum-littorale (C. psl.) complex have two sexes, mating-type plus (mt + ) and mating-type minus (mt - ). Conjugation between these two sexes is regulated by two sex pheromones, protoplast-release-inducing protein (PR-IP) and PR-IP Inducer, which are produced by mt + and mt - cells, respectively. PR-IP mediates the release of protoplasts from mt - cells during mating. In this study, we examined the mechanism of action of CpRLP1 (receptor-like protein 1), which was previously identified in a cDNA microarray analysis as one of the PR-IP-inducible genes. Using CRISPR/Cas9 technology, we generated CpRLP1 knockout mutants in mt - cells of the C. psl. complex. When the knockout mt - cells were mixed with wild-type mt + cells, conjugation was severely reduced. Many cells released protoplasts without pairing, suggesting a loss of synchronization between the two mating partners. Furthermore, the knockout mutants were hypersensitive to PR-IP. We conclude that CpRLP1 is a negative regulator of PR-IP that regulates the timing of protoplast release in conjugating C. psl. cells. As the first report of successful gene knockout in the class Charophyceae, this study provides a basis for research aimed at understanding the ancestral roles of genes that are indispensable for the development of land plants.

77 FR 28766 - Eighth Coast Guard District Annual Marine Events and Safety Zones

Science.gov (United States)

2012-05-16

.... SUMMARY: On March 1, 2012, the Coast Guard published a direct final rule, amending and updating its... effect as scheduled. The Coast Guard is also correcting two entries in this rule through technical... event's date, location, and the resulting special local regulation remain the same. The next occurrence...

30 CFR 57.12080 - Bare conductor guards.

Science.gov (United States)

2010-07-01

....12080 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND NONMETAL MINE SAFETY AND HEALTH SAFETY AND HEALTH STANDARDS-UNDERGROUND METAL AND NONMETAL MINES Electricity... conductors are less than 7 feet above the rail, they shall be guarded at all points where persons work or...

Virus-specific proteins in cells infected with tomato black ring nepovirus: evidence for proteolytic processing in vivo.

OpenAIRE

Demangeat, Gerard; Hemmer, O; Reinbolt, J; Mayo, M A; Fritsch, Coralie

1992-01-01

The synthesis of proteins encoded by the RNA of tomato black ring virus (TBRV) in vivo was studied in protoplasts by direct labelling with [35S]methionine, and in protoplasts and plants by immunoblotting experiments with specific antisera. Comparison of the proteins synthesized in infected and mock-inoculated protoplasts suggested that proteins of M(r) 120K, 90K, 80K, 57K and 46K were virus-specific. The proteins derived from the RNA-1-encoded polyprotein detected by immunoblotting were a sta...

Effects of ultraviolet radiation on microtubule organisation and morphogenesis in plants

International Nuclear Information System (INIS)

Staxen, I.

1994-09-01

The involvement of the cytoskeleton in the development of somatic embryos was studied in Larix x eurolepis. Protoplasts were isolated from both somatic embryo-regenerating and non-generating cultures and fractionated on a discontinuous Percoll density gradient, whereby a highly embryogenic protoplast fraction could be enriched. Protoplasts of two cell lines of Larix eurolepis, one with regenerating potential and one lacking this potential, were compared. In contrast to the non-regenerating line were a protoplast-like organisation of the cortical microtubules was maintained, re-organisation of this microtubular network occurred in the regenerable line after only three days of culture, indicating that organised growth was occurring. However, this early organisation of cortical microtubules may not always be a valid marker for regenerable and non-regenerable material. In order to investigate the effect of ultraviolet-B (UV-B, 280-320 nm) radiation on the microtubule cytoskeleton, protoplasts were isolated from leaves of Petunia hybrida and subjected to four different doses of UV-B radiation. The organisation of the microtubules and the progression of the cells through the cell cycle was observed at 0, 24, 48 and 72 h after irradiation. UV-B induced breaks in the cortical microtubules resulting in shorter fragments with increasing amounts of radiation. Also, the division of the protoplasts was delayed, which was related to the absence of an microtubule network. Whole Petunia plants were grown in growth chambers in the presence and absence of UV-B. The plants responded to UV-B with increased rates of CO 2 assimilation, a 60% increase in UV-screening compounds and the changes in the morphology of the leaves that were reflected in a 70-100% increase in leaf area and 20% decrease in leaf thickness. The microtubules of the epidermal cells was not affected by UV-B, nor was the number of epidermal cells (per unit area). The increase in leaf area in the UV-treated plants

Sperm competition in humans: mate guarding behavior negatively correlates with ejaculate quality.

Science.gov (United States)

Leivers, Samantha; Rhodes, Gillian; Simmons, Leigh W

2014-01-01

In species where females mate with multiple males, the sperm from these males must compete to fertilise available ova. Sexual selection from sperm competition is expected to favor opposing adaptations in males that function either in the avoidance of sperm competition (by guarding females from rival males) or in the engagement in sperm competition (by increased expenditure on the ejaculate). The extent to which males may adjust the relative use of these opposing tactics has been relatively neglected. Where males can successfully avoid sperm competition from rivals, one might expect a decrease in their expenditure on tactics for the engagement in sperm competition and vice versa. In this study, we examine the relationship between mate guarding and ejaculate quality using humans as an empirical model. We found that men who performed fewer mate guarding behaviors produced higher quality ejaculates, having a greater concentration of sperm, a higher percentage of motile sperm and sperm that swam faster and less erratically. These effects were found independent of lifestyle factors or factors related to male quality. Our findings suggest that male expenditure on mate guarding and on the ejaculate may represent alternative routes to paternity assurance in humans.

Elimination of Coast Guard plan review for non-critical engineering systems and cargo barges

Science.gov (United States)

1994-06-21

The purpose of this Circular is to publish policy that provides for the elimination of Coast Guard review and approval of certain engineering system drawings for all vessels as well as structural drawings for Coast Guard inspected non-self propelled ...

Double-blind evaluation of the DKL LifeGuard Model 2

International Nuclear Information System (INIS)

Murray, D.W.; Spencer, F.W.; Spencer, D.D.

1998-05-01

On March 20, 1998, Sandia National Laboratories performed a double-blind test of the DKL LifeGuard human presence detector and tracker. The test was designed to allow the device to search for individuals well within the product's published operational parameters. The Test Operator of the DKL LifeGuard was provided by the manufacturer and was a high-ranking member of DKL management. The test was developed and implemented to verify the performance of the device as specified by the manufacturer. The device failed to meet its published specifications and it performed no better than random chance

Relational Reasoning for Markov Chains in a Probabilistic Guarded Lambda Calculus

DEFF Research Database (Denmark)

Aguirre, Alejandro; Barthe, Gilles; Birkedal, Lars

2018-01-01

We extend the simply-typed guarded $\\lambda$-calculus with discrete probabilities and endow it with a program logic for reasoning about relational properties of guarded probabilistic computations. This provides a framework for programming and reasoning about infinite stochastic processes like Mar...... literature to justify better proof rules for relational reasoning about probabilistic expressions. We illustrate these benefits with a broad range of examples that were beyond the scope of previous systems, including shift couplings and lump couplings between random walks....

The study of the morphological features of autophagy as a type of programmed death of plant cells under the condition of bacterial infection

Directory of Open Access Journals (Sweden)

Сергій Іванович Шевченко

2016-09-01

Full Text Available The ultrastructure of the destruction of the plant cells protoplast is studied under the condition of bacterial infection. According to the autophagy processes in animal cells, the morphological ways of plant cells autophagy – vacuolization of cytoplasm, condensation and decondensation of the nuclear mass, multivesicular nucleation, phagophore expansion and macroautophagosome ripening, autophagolysosome formation by the way of tonoplast invagination, mitophagy phenomenon are determined. The places of the final degradation of the ruined cytoplasm in the vacuoles of destroyed cells are shown

78 FR 77359 - Eighth Coast Guard District Annual Safety Zones; New Year's Eve Celebration/City of Mobile...

Science.gov (United States)

2013-12-23

...-AA00 Eighth Coast Guard District Annual Safety Zones; New Year's Eve Celebration/City of Mobile; Mobile Channel; Mobile, AL AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard will enforce the City of Mobile New Year's Eve Celebration safety zone in the Mobile Channel, Mobile, AL from...

AlphaGUARD, the new reference for continuous radon monitoring in air, soil, gas, water and material; AlphaGUARD, die neue Referenz fuer die kontinuierliche Messung der Radonkonzentration in Luft, Boden, Wasser und Baumaterial

Energy Technology Data Exchange (ETDEWEB)

Roessler, F.; Buerkin, W. [Saphymo GmbH, Frankfurt am Main (Germany); Villert, J. [Bertin Technologies, Montigny (France)

2016-07-01

The company Saphymo GmbH has more than 25 years of experience in the field of radon measurement. More than 20 years ago Saphymo developed the professional and robust radon monitor AlphaGUARD, quickly recognized as a standard for reliable and continuous measurements of the radon concentration. Today AlphaGUARD is internationally established as the reference in radon measurement. Following up on this success story the new generation of AlphaGUARD can now be presented. Based on the excellent measurement characteristics of its predecessor the new AlphaGUARD combines the well-proven principle of the pulse ionisation chamber with new and additional features. The robust housing is oriented on the well-proven design of the predecessor and includes now an integrated flow controlled and powerful pump. The instrument can be operated in flow as well as in diffusion mode (without pump). Via the new large display and the intuitive menu navigation all measurement data can be retrieved. The presentation of time series in charts is possible as well as the parametrisation of the instrument. A wide range of accessories, developed in cooperation with various radon experts of universities and laboratories, enables the user a varied and flexible application of the AlphaGUARD: Measurement of the radon concentration in air (radon, thoron, radon progenies), in water (sampling and time resolved measurements) and in soil (soil gas measurements, exhalation measurements), emanation measurements from material, multi spot measurement, online measurement with remote data transmission via Ethernet/DSL, Bluetooth, Wi-Fi, GPRS/3G or satellite. Due to its high sensitivity and its fast and linear response over a large measuring range the AlphaGUARD is excellently suited for calibration laboratories. Furthermore the AlphaGUARD enables ideal prerequisites for field applications: robust housing for operations under harsh conditions, long battery life for the measurement at any location, low

METACASPASE9 modulates autophagy to confine cell death to the target cells during Arabidopsis vascular xylem differentiation

Directory of Open Access Journals (Sweden)

Sacha Escamez

2016-02-01

Full Text Available We uncovered that the level of autophagy in plant cells undergoing programmed cell death determines the fate of the surrounding cells. Our approach consisted of using Arabidopsis thaliana cell cultures capable of differentiating into two different cell types: vascular tracheary elements (TEs that undergo programmed cell death (PCD and protoplast autolysis, and parenchymatic non-TEs that remain alive. The TE cell type displayed higher levels of autophagy when expression of the TE-specific METACASPASE9 (MC9 was reduced using RNAi (MC9-RNAi. Misregulation of autophagy in the MC9-RNAi TEs coincided with ectopic death of the non-TEs, implying the existence of an autophagy-dependent intercellular signalling from within the TEs towards the non-TEs. Viability of the non-TEs was restored when AUTOPHAGY2 (ATG2 was downregulated specifically in MC9-RNAi TEs, demonstrating the importance of autophagy in the spatial confinement of cell death. Our results suggest that other eukaryotic cells undergoing PCD might also need to tightly regulate their level of autophagy to avoid detrimental consequences for the surrounding cells.

Living target of Ce(III) action on horseradish cells: proteins on/in cell membrane.

Science.gov (United States)

Yang, Guangmei; Sun, Zhaoguo; Lv, Xiaofen; Deng, Yunyun; Zhou, Qing; Huang, Xiaohua

2012-12-01

Positive and negative effects of rare earth elements (REEs) in life have been reported in many papers, but the cellular mechanisms have not been answered, especially the action sites of REEs on plasma membrane are unknown. Proteins on/in the plasma membrane perform main functions of the plasma membrane. Cerium (Ce) is the richest REEs in crust. Thus, the interaction between Ce(III) and the proteins on/in the plasma membrane, the morphology of protoplast, and the contents of nutrient elements in protoplast of horseradish were investigated using the optimized combination of the fluorescence microscopy, fluorescence spectroscopy, circular dichroism, scanning electron microscopy, and X-ray energy dispersive spectroscopy. It was found that Ce(III) at the low concentrations (10, 30 μM) could interact with proteins on/in the plasma membrane of horseradish, leading to the improvement in the structure of membrane proteins and the plasma membrane, which accelerated the intra-/extra-cellular substance exchange and further promoted the development of cells. When horseradish was treated with Ce(III) at the high concentrations (60, 80 μM), Ce(III) also could interact with the proteins on/in the plasma membrane of horseradish, leading to the destruction in the structure of membrane proteins and the plasma membrane. These effects decelerated the intra-/extra-cellular substance exchange and further inhibited the development of cells. Thus, the interaction between Ce(III) and proteins on/in the plasma membrane in plants was an important reason of the positive and negative effects of Ce(III) on plants. The results would provide some references for understanding the cellular effect mechanisms of REEs on plants.

Correlation Investigation as to the Mental Health of the Prison Guard in Xi'an Police Station

Institute of Scientific and Technical Information of China (English)

Feng Xue

2005-01-01

Based on the symptom check list 90,EPQ scale and the simplified coping style questionaire,the author made a relative study on psychological health,personality character and coping style of 267 prison guard of police bureau in Xi'an city .The study shows this: 　　①The mental health,personality character and coping style could be effected by many factors,such as gender,age,educational background,income,origin,satisfactory degree of working enviroment and some others.The mental health,personality character and coping style could effect each other.②The psychological health state of 267 prison guard is much worse than that of domestic common people.③The personality character of 267 prison guard shows much higher hostility and lower nervous than that of domestic common people.④The coping style of 267 prison guard often take positive coping styles.⑤Psychological health,personality character and coping style of prison guard are closely related.

Redesign of the Advanced Education processes in the United States Coast Guard

OpenAIRE

Johnson, Lamar V.; Sanders, Marc F.

1999-01-01

The processes used in the operation of the Coast Guard Advanced Education Program have evolved as most business processes that were developed prior to the introduction of information technology. These processes include the selection, management, assignment and tracking of advanced education students. These processes are still fully dependent on physical files and the mail system. The Coast Guard has an information technology infrastructure that supports better processes, however it is not bei...

Advertisement call and tadpole morphology of the clutch-guarding ...

African Journals Online (AJOL)

Advertisement call and tadpole morphology of the clutch-guarding frog Mantidactylus argenteus from eastern ... We provide the first descriptions of the tadpole and advertisement call of Mantidactylus argenteus. ... AJOL African Journals Online.

Probabilistic Role Models and the Guarded Fragment

DEFF Research Database (Denmark)

Jaeger, Manfred

2004-01-01

We propose a uniform semantic framework for interpreting probabilistic concept subsumption and probabilistic role quantification through statistical sampling distributions. This general semantic principle serves as the foundation for the development of a probabilistic version of the guarded fragm...... fragment of first-order logic. A characterization of equivalence in that logic in terms of bisimulations is given....

Probabilistic role models and the guarded fragment

DEFF Research Database (Denmark)

Jaeger, Manfred

2006-01-01

We propose a uniform semantic framework for interpreting probabilistic concept subsumption and probabilistic role quantification through statistical sampling distributions. This general semantic principle serves as the foundation for the development of a probabilistic version of the guarded fragm...... fragment of first-order logic. A characterization of equivalence in that logic in terms of bisimulations is given....

Personal Staff - Joint Staff - The National Guard

Science.gov (United States)

the ARNG Deputy Director of the ARNG Chief of Staff of the ARNG Command Chief Warrant Officer of the Site Maintenance Battle Focused Training Strategy Battle Staff Training Resources News Publications March Today in Guard History Leadership CNGB VCNGB SEA DANG DARNG Joint Staff J-1 J-2 J-3 J-4 J-5 J-6 J

Use of ISEM in studying the impact of guard tactics on facility safeguards system effectiveness

International Nuclear Information System (INIS)

Engi, D.; Boozer, D.D.

1977-07-01

The Insider Safeguards Effectiveness Model (ISEM) is a stochastic, discrete event, Monte-Carlo Simulation Model used to assess the effectiveness of physical protection systems for facilities which store, process, or use SNM. ISEM simulates the interaction of a group of insiders. The sensor control and alarm locations are correlated with the authorized access areas of the insider(s) and, if the insider has the appropriate access, the probability of an insider successfully defeating a sensor control or alarm is computed based upon the surveillance subsystems and the insider's attributes. Following an alarm and an assessment, actions are initiated by the safeguards system. These actions typically involve dispatching guards to specific locations within the facility. Since the specific guard responses must be predetermined, a safeguards system should be evolved from a consideration of a wide spectrum of feasible adversary strategies. The sensitivity of safeguards system effectiveness to a variety of guard tactics is explored in this paper. The evolution of comprehensive guard tactics for protecting a hypothetical facility is demonstrated. Attention is focused on the potential threat posed by insiders and the necessity of well conceived guard tactics in dealing with this threat

ISOLASI DAN KULTUR PROTOPLAS RUMPUT LAUT Kappaphycus alvarezii DI LABORATORIUM

Directory of Open Access Journals (Sweden)

Emma Suryati

2007-12-01

Full Text Available Isolasi protoplas rumput laut K. alvarezii, telah dilakukan dalam rangka penyiapan protoplas untuk penyilangan melalui fusi protoplas. Metode yang digunakan antara lain melalui cara kimia dengan melisis tallus rumput laut dengan campuran enzim komersial, kemudian enzim yang berasal dari viscera keong mas baik yang segar maupun yang beku, dengan media kultur yang digunakan pada pemeliharaan makro algae antara lain Conwy, PES, dan air laut steril. Tallus rumput laut yang digunakan berasal dari bagian pangkal, tengah dan ujung. Protoplas yang hidup diuji menggunakan evans blue 0,1%, hormon perangsang tumbuh yang digunakan pada media pertumbuhan antara lain auxin, IAA, dan Kinetin. Pengamatan dilakukan terhadap jumlah protoplas hidup, pertumbuhan, dan sintasan.  Hasil percobaan memperlihatkan bahwa enzim yang paling baik digunakan adalah campuran enzim komersial dengan media kultur Conwy dengan jumlah protoplas mencapai 19,8 x 106 sel/mL, bagian tallus yang paling baik adalah bagian pangkal berkisar antara 8,1x106 hingga 18,8 x 106 sel/ mL.  Perangsang tumbuh yang paling baik adalah auxin. Filamen terbentuk setelah 5 hari dengan fotoperiod L:D=12:12. Isolation of seaweed’s protoplast Kappaphycus alvarezii had been done to provide protoplast for crossbreeding purpose by protoplast fusion. The method was chemically done by lyses of tallus used commercial enzyme mixture, enzyme from viscera of snail both fresh and frozen, culture media were Conwy (CW, PES, and sterile sea water (SSW which were used to maintain the macro algae. Part of used tallus were upper, middle and tip of tallus. The viable protoplast was examined by using 0.1% evans blue and the growth-stimulating hormone were auxin, IAA, and Kinetin. Observation was concerned to the amount of viable protoplast, the growth, and the long live. Result showed that the best enzyme was commercial enzyme mixture with Conwy as the best culture media, provided protoplast until 19.8 x 106 cell/mL. The

78 FR 33216 - Temporary Change of Dates for Recurring Marine Event in the Fifth Coast Guard District, Mattaponi...

Science.gov (United States)

2013-06-04

... 1625-AA08 Temporary Change of Dates for Recurring Marine Event in the Fifth Coast Guard District, Mattaponi Drag Boat Race, Mattaponi River; Wakema, VA AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard is establishing a temporary change to the enforcement period of special local...

76 FR 30827 - Temporary Change of Dates for Recurring Marine Event in the Fifth Coast Guard District; Elizabeth...

Science.gov (United States)

2011-05-27

...-AA08 Temporary Change of Dates for Recurring Marine Event in the Fifth Coast Guard District; Elizabeth River, Norfolk, VA AGENCY: Coast Guard, DHS. ACTION: Temporary final rule. SUMMARY: The Coast Guard will... various river boat races and a parade during the ``35th Annual Norfolk Harborfest Celebration.'' Special...

Assessment of private security guards by Suicide Probability Scale and Brief Symptom Inventory.

Science.gov (United States)

Dogan, Bulent; Canturk, Gurol; Canturk, Nergis; Guney, Sevgi; Özcan, Ebru

2016-01-01

The aim of the present study was to investigate the influence of suicide probability and relevant sociodemographic features and to provide information for preventing suicide in private security guards working under the stressful conditions and continuous exposure to the negative and traumatic life events. 200 private security guards and 200 personnels of Ankara University participated in the study. A sociodemographic information questionnaire, the Suicide Probability Scale (SPS) and the Brief Symptom Inventory (BSI) were used to collect the data. Gender, marital status, income, religious beliefs, experiencing a life-threatening situation, history of a suicide attempt, smoking and not having a chronic disease caused statistically significant differences in the scores for SPS between the private security guards group and the controls. Moreover there was a statistically significant positive correlation between the total scores of the subscales of SPS and the total scores of BSI. Like police officers and gendarmes, private security guards are at high risk of committing and attempting suicide because of being at stressful work settings and also suffering from secondary trauma. It is required that they should be aware of their tendency to commit suicide and have regular psychiatric screenings.

The National Guard Ballistic Missile Defense Mission minutemen at the Orgital Plane

OpenAIRE

Trenary, Ralph Hiram

2004-01-01

Approved for public release; distribution is unlimited This thesis examines the decision to assign the Ballistic Missile Defense mission to units of the Colorado and Alaska National Guard. The history of the Nike Ajax, Nike Hercules, Sentinel and Safeguard programs are examined to identify the origins of support for this decision. First-hand sources provide evidence that the National Guard performance in the Nike air defense program is a record of parity and some superiority to equivalent ...

Diagnostic double guarded low-volume uterine lavage in mares

DEFF Research Database (Denmark)

Christoffersen, Mette; Brandis, Louise; Samuelsson, Julia

2014-01-01

Endometritis constitutes a major problem in the management of broodmares; hence diagnostic tests with a high sensitivity and specificity are desired. We hypothesize that a double guarded uterine flush technique for bacterial culture and cytology is comparable to standard diagnostic tests, the end......Endometritis constitutes a major problem in the management of broodmares; hence diagnostic tests with a high sensitivity and specificity are desired. We hypothesize that a double guarded uterine flush technique for bacterial culture and cytology is comparable to standard diagnostic tests...... (lavage: 11%, swab: 8%, biopsy: 7%) (positive bacterial growth > 4 Colony forming units (CFU)). Positive cytology was less likely to occur when E. coli was isolated from the diagnostic tests compared to the growth of β-hemolytic streptococci. Isolation of pathogens from uterine samples was highly...

Specific recognition of reproductive parasite workers by nest-entrance guards in the bumble bee Bombus terrestris.

Science.gov (United States)

Blacher, Pierre; Boreggio, Laurie; Leroy, Chloé; Devienne, Paul; Châline, Nicolas; Chameron, Stéphane

2013-12-10

The impact of social parasites on their hosts' fitness is a strong selective pressure that can lead to the evolution of adapted defence strategies. Guarding the nest to prevent the intrusion of parasites is a widespread response of host species. If absolute rejection of strangers provides the best protection against parasites, more fine-tuned strategies can prove more adaptive. Guarding is indeed costly and not all strangers constitute a real threat. That is particularly true for worker reproductive parasitism in social insects since only a fraction of non-nestmate visitors, the fertile ones, can readily engage in parasitic reproduction. Guards should thus be more restrictive towards fertile than sterile non-nestmate workers. We here tested this hypothesis by examining the reaction of nest-entrance guards towards nestmate and non-nestmate workers with varying fertility levels in the bumble bee Bombus terrestris. Because social recognition in social insects mainly relies on cuticular lipids (CLs), chemical analysis was also conducted to examine whether workers' CLs could convey the relevant information upon which guards could base their decision. We thus aimed to determine whether an adapted defensive strategy to worker reproductive parasitism has evolved in B. terrestris colonies. Chemical analysis revealed that the cuticular chemical profiles of workers encode information about both their colony membership and their current fertility, therefore providing potential recognition cues for a suitable adjustment of the guards' defensive decisions. We found that guards were similarly tolerant towards sterile non-nestmate workers than towards nestmate workers. However, as predicted, guards responded more aggressively towards fertile non-nestmates. Our results show that B. terrestris guards discriminate non-nestmates that differ in their reproductive potential and respond more strongly to the individuals that are a greatest threat for the colony. Cuticular hydrocarbons are

Coloured leg bands affect male mate-guarding behaviour in the bluethroat

Science.gov (United States)

Johnsen; Lifjeld; Rohde

1997-07-01

Artificial traits such as coloured leg bands may affect an individual's mating success, as shown for some birds. One explanation is that colour-matching with a sexual ornament affects the individual's sexual attractiveness. This study reports a colour-band experiment with free-living bluethroats, Luscinia s. svecicaa species where males have a distinct blue and chestnut throat and upper breast. There was no apparent difference in pairing success between males with ornament-matching colour bands (blue and orange) and males with non-ornamental colour bands. However, males with ornamental bands guarded their mates less intensely and spent more time singing, performing song flights and intruding into neighbours' territories than males with non-ornamental bands. We conclude that colour bands affect the trade-off between mate guarding and advertisement behaviour in a way that is consistent with the hypothesis that bands with ornamental colours improve a male's attractiveness. The results are in concordance with a previous study of the same population, showing that males with experimentally reduced attractiveness guarded their mates more closely and advertised less for additional mates, than non-manipulated males.

Multi-species consumer jams and the fall of guarded corals to crown-of-thorns seastar outbreaks

Science.gov (United States)

Kayal, Mohsen; Ballard, Jane; Adjeroud, Mehdi

2018-01-01

Outbreaks of predatory crown-of-thorns seastars (COTS) can devastate coral reef ecosystems, yet some corals possess mutualistic guardian crabs that defend against COTS attacks. However, guarded corals do not always survive COTS outbreaks, with the ecological mechanisms sealing the fate of these corals during COTS infestations remaining unknown. In August 2008 in Moorea (17.539° S, 149.830° W), French Polynesia, an unusually dense multi-species aggregation of predators was observed feeding upon guarded corals following widespread coral decline due to COTS predation. Concurrent assaults from these amplified, mixed-species predator guilds likely overwhelm mutualistic crab defense, ultimately leading to the fall of guarded corals. Our observations indicate that guarded corals can sustain devastating COTS attacks for an extended duration, but eventually concede to intensifying assaults from diverse predators that aggregate in high numbers as alternative prey decays. The fall of guarded corals is therefore suggested to be ultimately driven by an indirect trophic cascade that leads to amplified attacks from diverse starving predators following prey decline, rather than COTS assaults alone. PMID:29487739

78 FR 40000 - Eighth Coast Guard District Annual Safety Zones; Riverfront Independence Festival Fireworks; Ohio...

Science.gov (United States)

2013-07-03

...-AA00 Eighth Coast Guard District Annual Safety Zones; Riverfront Independence Festival Fireworks; Ohio.... SUMMARY: The Coast Guard will enforce a Safety Zone for the Riverfront Independence Festival Fireworks on... navigable waters during the Riverfront Independence Festival Fireworks. During the enforcement period, in...

Major benefits of guarding behavior in subsocial bees: implications for social evolution

OpenAIRE

Mik?t, Michael; ?ern?, Kate?ina; Straka, Jakub

2016-01-01

Abstract Parental care is a behavior that increases the growth and survival of offspring, often at a cost to the parents' own survival and/or future reproduction. In this study, we focused on nest guarding, which is one of the most important types of extended parental care; we studied this behavior in two solitary bee species of the genus Ceratina with social ancestors. We performed the experiment of removing the laying female, who usually guards the nest after completing its provisioning, to...

Refining types using type guards in TypeScript

NARCIS (Netherlands)

de Wolff, Ivo Gabe; Hage, J.

2017-01-01

We discuss two adaptations of the implementation of type guards and narrowing in the TypeScript compiler. The first is an improvement on the original syntax-directed implementation, and has now replaced the original one in the TypeScript compiler. It is specifically suited for the scenario in which

EPA Region 1 Coast Guard Jurisdictional Boundary - Polygons

Science.gov (United States)

Jurisdictional boundary between EPA and Coast Guard for EPA Region I. Created from 1:100000 USGS DLGs with greater detail drawn from 1:24000 commercial street data for Region I.This layer is used to determine which agency will be reponsible in the event of an oil spill.

EPA Region 1 Coast Guard Jurisdictional Boundary - Arcs

Science.gov (United States)

Jurisdictional boundary between EPA and Coast Guard for EPA Region I. Created from 1:100000 USGS DLGs with greater detail drawn from 1:24000 commercial street data for Region I.This layer is used to determine which agency will be reponsible in the event of an oil spill.

Tricking the guard: exploiting plant defense for disease susceptibility.

Science.gov (United States)

Lorang, J; Kidarsa, T; Bradford, C S; Gilbert, B; Curtis, M; Tzeng, S-C; Maier, C S; Wolpert, T J

2012-11-02

Typically, pathogens deploy virulence effectors to disable defense. Plants defeat effectors with resistance proteins that guard effector targets. We found that a pathogen exploits a resistance protein by activating it to confer susceptibility in Arabidopsis. The guard mechanism of plant defense is recapitulated by interactions among victorin (an effector produced by the necrotrophic fungus Cochliobolus victoriae), TRX-h5 (a defense-associated thioredoxin), and LOV1 (an Arabidopsis susceptibility protein). In LOV1's absence, victorin inhibits TRX-h5, resulting in compromised defense but not disease by C. victoriae. In LOV1's presence, victorin binding to TRX-h5 activates LOV1 and elicits a resistance-like response that confers disease susceptibility. We propose that victorin is, or mimics, a conventional pathogen virulence effector that was defeated by LOV1 and confers virulence to C. victoriae solely because it incites defense.

20 CFR 1002.306 - Is a National Guard civilian technician considered a State or Federal employee for purposes of...

Science.gov (United States)

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Is a National Guard civilian technician... a National Guard civilian technician considered a State or Federal employee for purposes of USERRA? A National Guard civilian technician is considered a State employee for USERRA purposes, although he...

U.S. Coast Guard (USCG) Maritime Differential GPS (DGPS) Locations

Data.gov (United States)

Department of Homeland Security — NAVCEN operates the Coast Guard Maritime Differential GPS (DGPS) Service and the developing Nationwide DGPS Service (NDGPS), consisting of two control centers and 86...

Examining Ankle-Joint Laxity Using 2 Knee Positions and With Simulated Muscle Guarding.

Science.gov (United States)

Hanlon, Shawn; Caccese, Jaclyn; Knight, Christopher A; Swanik, Charles Buz; Kaminski, Thomas W

2016-02-01

Several factors affect the reliability of the anterior drawer and talar tilt tests, including the individual clinician's experience and skill, ankle and knee positioning, and muscle guarding. To compare gastrocnemius activity during the measurement of ankle-complex motion at different knee positions, and secondarily, to compare ankle-complex motion during a simulated trial of muscle guarding. Cross-sectional study. Research laboratory. Thirty-three participants aged 20.2 ± 1.7 years were tested. The ankle was loaded under 2 test conditions (relaxed, simulated muscle guarding) at 2 knee positions (0°, 90° of flexion) while gastrocnemius electromyography (EMG) activity was recorded. Anterior displacement (mm), inversion-eversion motion (°), and peak EMG amplitude values of the gastrocnemius (μV). Anterior displacement did not differ between the positions of 0° and 90° of knee flexion (P = .193). Inversion-eversion motion was greater at 0° of knee flexion compared with 90° (P ankle laxity at the 2 most common knee positions for anterior drawer testing; however, talar tilt testing may be best performed with the knee in 0° of knee flexion. Finally, our outcomes from the simulated muscle-guarding condition suggest that clinicians should use caution and be aware of reduced perceived laxity when performing these clinical examination techniques immediately postinjury.

Small incision guarded hydroaspiration of iris lesions.

Science.gov (United States)

Singh, Arun D

2017-11-01

To describe the technique and results of a minimally invasive surgical technique for resection of small iris lesions. Consecutive case series of 22 patients with localised, small iris lesions that were resected using the described surgical technique that composed of multiple, small corneal incisions created to allow for internal iris resection with 23-gauge horizontal vitrectomy scissors, followed by guarded tumour aspiration through a clear plastic tubing (diameter 3.5 mm) primed with viscoelastic agent. The mean largest basal diameter was 3.0 mm (range 1.5-5.0 mm; median 3.0 mm) and mean thickness was 1.3 mm (range 0.5-2.5 mm; median 1.0 mm). Use of multiple (2-4) small corneal incisions (range 2.0-3.0 mm; mean 2.8 mm) allowed reduced postoperative morbidity (significant hyphema (0%), hypotony (0%), wound leak (0%), >2 line change in best corrected visual acuity at postoperative 1 week (4.5%) and mean corneal astigmatism of 1.0 D (range 0.14-2.99 D; median 0.8 D) at postoperative 4-12 weeks. The tumour could be resected with clear surgical margins in all neoplastic cases (benign (2), borderline (1) and malignant (16)). Local recurrence or metastases were not observed in any melanoma case over a mean follow-up of 33.0 months (range 1.0-90.0 months; median 33.5 months). Small incision guarded hydroaspiration is a minimally invasive surgical technique for resection of select small iris lesions. Use of multiple small corneal incisions avoids morbidity associated with a single large corneoscleral incision, and use of guarded aspiration may eliminate the risk of wound contamination by the malignant tumour. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

The Arabidopsis arc5 and arc6 mutations differentially affect plastid morphology in pavement and guard cells in the leaf epidermis.

Science.gov (United States)

Fujiwara, Makoto T; Yasuzawa, Mana; Kojo, Kei H; Niwa, Yasuo; Abe, Tomoko; Yoshida, Shigeo; Nakano, Takeshi; Itoh, Ryuuichi D

2018-01-01

Chloroplasts, or photosynthetic plastids, multiply by binary fission, forming a homogeneous population in plant cells. In Arabidopsis thaliana, the division apparatus (or division ring) of mesophyll chloroplasts includes an inner envelope transmembrane protein ARC6, a cytoplasmic dynamin-related protein ARC5 (DRP5B), and members of the FtsZ1 and FtsZ2 families of proteins, which co-assemble in the stromal mid-plastid division ring (FtsZ ring). FtsZ ring placement is controlled by several proteins, including a stromal factor MinE (AtMinE1). During leaf mesophyll development, ARC6 and AtMinE1 are necessary for FtsZ ring formation and thus plastid division initiation, while ARC5 is essential for a later stage of plastid division. Here, we examined plastid morphology in leaf epidermal pavement cells (PCs) and stomatal guard cells (GCs) in the arc5 and arc6 mutants using stroma-targeted fluorescent proteins. The arc5 PC plastids were generally a bit larger than those of the wild type, but most had normal shapes and were division-competent, unlike mutant mesophyll chloroplasts. The arc6 PC plastids were heterogeneous in size and shape, including the formation of giant and mini-plastids, plastids with highly developed stromules, and grape-like plastid clusters, which varied on a cell-by-cell basis. Moreover, unique plastid phenotypes for stomatal GCs were observed in both mutants. The arc5 GCs rarely lacked chlorophyll-bearing plastids (chloroplasts), while they accumulated minute chlorophyll-less plastids, whereas most GCs developed wild type-like chloroplasts. The arc6 GCs produced large chloroplasts and/or chlorophyll-less plastids, as previously observed, but unexpectedly, their chloroplasts/plastids exhibited marked morphological variations. We quantitatively analyzed plastid morphology and partitioning in paired GCs from wild-type, arc5, arc6, and atminE1 plants. Collectively, our results support the notion that ARC5 is dispensable in the process of equal division

Task force St. Bernard: operational issues and medical management of a National Guard disaster response operation.

Science.gov (United States)

Bonnett, Carl J; Schock, Tony R; McVaney, Kevin E; Colwell, Christopher B; Depass, Christopher

2007-01-01

After Hurricane Katrina struck the Gulf Coast of the United States on 29 August 2005, it became obvious that the country was facing an enormous national emergency. With local resources overwhelmed, governors across the US responded by deploying thousands of National Guard soldiers and airmen. The National Guard has responded to domestic disasters due to natural hazards since its inception, but an event with the magnitude of Hurricane Katrina was unprecedented. The deployment of >900 Army National Guard soldiers to St. Bernard Parish, Louisiana in the aftermath of the Hurricane was studied to present some of the operational issues involved with providing medical support for this type of operation. In doing so, the authors attempt to address some of the larger issues of how the National Guard can be incorporated into domestic disaster response efforts. A number of unforeseen issues with regards to medical operations, medical supply, communication, preventive medicine, legal issues, and interactions with civilians were encountered and are reviewed. A better understanding of the National Guard and how it can be utilized more effectively in future disaster response operations can be developed.

77 FR 35855 - Safety Zone, Temporary Change for Recurring Fifth Coast Guard District Fireworks Displays...

Science.gov (United States)

2012-06-15

... 1625-AA00 Safety Zone, Temporary Change for Recurring Fifth Coast Guard District Fireworks Displays; Northwest Harbor (East Channel) and Tred Avon River, MD AGENCY: Coast Guard, DHS. ACTION: Temporary final... Channel), Patapsco River, and Tred Avon River during the event. DATES: This rule is effective from 8 p.m...

[Burnout Syndrome in Prison Guards, Bucaramanga, Colombia, 2013].

Science.gov (United States)

Rodríguez Amaya, Reynaldo Mauricio

2014-01-01

Burnout syndrome is defined as a chronic stress response and sustained work environment, characterized by emotional exhaustion, depersonalization and personal accomplishment. Most studies have included teaching and health personnel. However, there are limitations in the development of studies on prison guards to observe this problem. This study provides some characteristics of prison guards and seeks to highlight the behavior of the constructs of burnout according to demographic and occupational profile. This was a descriptive cross-sectional study on 111 prison guards in the city of Bucaramanga. The Maslach Burnout Inventory was applied, as well as taking ethical considerations into account. A univariate and bivariate analysis was performed on the data. The mean age was 35.8 years, 76% had more than 10 years work experience, 24% have some additional academic activities at work. Only 3.7% had burnout syndrome as such. In emotional exhaustion, 25.2% are classified as high risk, 30% in depersonalization, and 16% personal accomplishment. Within the personal achievement, the group with more than 20 years experience were those who had a critical score (ANOVA χ(2), P=.002). Of those who had a risk score for depersonalization, 67% had worked more than 240 extra hours a month, with a crude odds ratio (OR)=4.66 (P=.033) and age-adjusted OR=4.65 (P=.035). Burnout in this occupational group was not as prevalent as in that found in other publications. In turn, of the three constructs, the most significant was depersonalization and no notable emotional burnout was observed. The most significant work variables were the number of hours worked and work experience. Copyright © 2013 Asociación Colombiana de Psiquiatría. Publicado por Elsevier España. All rights reserved.

National Guard Engagement in the Pacific: No Threat to Security

National Research Council Canada - National Science Library

Bour, Mark

2002-01-01

This thesis evaluates recent decisions to expand the National Guard (NGE) State Partnership Program into the Asia-Pacific region and examines potential effects this expansion will have on the new partners created...

Stomatal responses to flooding of the intercellular air spaces suggest a vapor-phase signal between the mesophyll and the guard cells.

Science.gov (United States)

Sibbernsen, Erik; Mott, Keith A

2010-07-01

Flooding the intercellular air spaces of leaves with water was shown to cause rapid closure of stomata in Tradescantia pallida, Lactuca serriola, Helianthus annuus, and Oenothera caespitosa. The response occurred when water was injected into the intercellular spaces, vacuum infiltrated into the intercellular spaces, or forced into the intercellular spaces by pressurizing the xylem. Injecting 50 mm KCl or silicone oil into the intercellular spaces also caused stomata to close, but the response was slower than with distilled water. Epidermis-mesophyll grafts for T. pallida were created by placing the epidermis of one leaf onto the exposed mesophyll of another leaf. Stomata in these grafts opened under light but closed rapidly when water was allowed to wick between epidermis and the mesophyll. When epidermis-mesophyll grafts were constructed with a thin hydrophobic filter between the mesophyll and epidermis stomata responded normally to light and CO(2). These data, when taken together, suggest that the effect of water on stomata is caused partly by dilution of K(+) in the guard cell and partly by the existence of a vapor-phase signal that originates in the mesophyll and causes stomata to open in the light.

Monitoring reactive oxygen species formation and localisation in living cells by use of the fluorescent probe CM-H(2)DCFDA and confocal laser microscopy

DEFF Research Database (Denmark)

Kristiansen, Kim Anker; Jensen, Poul Erik; Møller, Ian Max

2009-01-01

Reactive oxygen species (ROS) develop as a consequence of wounding, light stress and chemical imbalances but act also as signals in living cells. The integrity of cells is seriously endangered, if ROS cannot be controlled by scavenging molecules and other repair mechanisms of the cell. For studying...... ROS development and signalling under stress, a reliable indicator is needed. We have tested the ROS sensitive dye 5-(and-6) chloromethyl-2',7' dichlorodihydrofluorescein diacetate acetyl ester (CM-H(2)DCFDA) using onion bulb scale and leaf epidermis as well as Arabidopsis leaves and protoplasts. ROS...

Finding Their Way Back In: Family Reintegration Following Guard Deployment.

Science.gov (United States)

Messecar, Deborah C

2017-03-01

The aim of this study was to describe deployed National Guard members' and their families' perceptions of their experience with family reintegration, and the causes and conditions of challenges reintegration presents after deployment. A total of 26 National Guard members and 19 family members participated in individual (n = 22), couples (n = 6), or focus group (n = 17) interviews. In-depth interviews were used to assess needs and maximize input from military families regarding deployment-related experiences and reintegration issues. Qualitative coding and analysis of data were completed using NVivo. Finding their way back in is the key process that the military members must complete to successfully reestablish their desired social connections with the family and reclaim their place within the family. Several conditions shape the degree of challenges with reintegration that veterans and their family will encounter. These include preparation for deployment, length and type of deployment, communication during deployment, and finally, awareness of how deployment changes the military member and the family. Support resources dedicated to providing National Guard members and their families with assistance in preparing for deployments and educating them about the importance of communication during deployment should be maintained and expanded. Broader educational efforts that increase awareness of what to expect regarding how deployment changes the military member and the family are needed. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

AlphaGUARD, the new reference for continuous radon monitoring in air, soil, gas, water and material

International Nuclear Information System (INIS)

Roessler, F.; Buerkin, W.; Villert, J.

2016-01-01

The company Saphymo GmbH has more than 25 years of experience in the field of radon measurement. More than 20 years ago Saphymo developed the professional and robust radon monitor AlphaGUARD, quickly recognized as a standard for reliable and continuous measurements of the radon concentration. Today AlphaGUARD is internationally established as the reference in radon measurement. Following up on this success story the new generation of AlphaGUARD can now be presented. Based on the excellent measurement characteristics of its predecessor the new AlphaGUARD combines the well-proven principle of the pulse ionisation chamber with new and additional features. The robust housing is oriented on the well-proven design of the predecessor and includes now an integrated flow controlled and powerful pump. The instrument can be operated in flow as well as in diffusion mode (without pump). Via the new large display and the intuitive menu navigation all measurement data can be retrieved. The presentation of time series in charts is possible as well as the parametrisation of the instrument. A wide range of accessories, developed in cooperation with various radon experts of universities and laboratories, enables the user a varied and flexible application of the AlphaGUARD: Measurement of the radon concentration in air (radon, thoron, radon progenies), in water (sampling and time resolved measurements) and in soil (soil gas measurements, exhalation measurements), emanation measurements from material, multi spot measurement, online measurement with remote data transmission via Ethernet/DSL, Bluetooth, Wi-Fi, GPRS/3G or satellite. Due to its high sensitivity and its fast and linear response over a large measuring range the AlphaGUARD is excellently suited for calibration laboratories. Furthermore the AlphaGUARD enables ideal prerequisites for field applications: robust housing for operations under harsh conditions, long battery life for the measurement at any location, low

Mixing of maize and wheat genomic DNA by somatic hybridization in regenerated sterile maize plants.

Science.gov (United States)

Szarka, B.; Göntér, I.; Molnár-Láng, M.; Mórocz, S.; Dudits, D.

2002-07-01

Intergeneric somatic hybridization was performed between albino maize ( Zea mays L.) protoplasts and mesophyll protoplasts of wheat ( Triticum aestivum L.) by polyethylene glycol (PEG) treatments. None of the parental protoplasts were able to produce green plants without fusion. The maize cells regenerated only rudimentary albino plantlets of limited viability, and the wheat mesophyll protoplasts were unable to divide. PEG-mediated fusion treatments resulted in hybrid cells with mixed cytoplasm. Six months after fusion green embryogenic calli were selected as putative hybrids. The first-regenerates were discovered as aborted embryos. Regeneration of intact, green, maize-like plants needed 6 months of further subcultures on hormone-free medium. These plants were sterile, although had both male and female flowers. The cytological analysis of cells from callus tissues and root tips revealed 56 chromosomes, but intact wheat chromosomes were not observed. Using total DNA from hybrid plants, three RAPD primer combinations produced bands resembling the wheat profile. Genomic in situ hybridization (GISH) using total wheat DNA as a probe revealed the presence of wheat DNA islands in the maize chromosomal background. The increased viability and the restored green color were the most-significant new traits as compared to the original maize parent. Other intermediate morphological traits of plants with hybrid origin were not found.

National machine guarding program: Part 1. Machine safeguarding practices in small metal fabrication businesses

OpenAIRE

Parker, David L.; Yamin, Samuel C.; Brosseau, Lisa M.; Xi, Min; Gordon, Robert; Most, Ivan G.; Stanley, Rodney

2015-01-01

Background Metal fabrication workers experience high rates of traumatic occupational injuries. Machine operators in particular face high risks, often stemming from the absence or improper use of machine safeguarding or the failure to implement lockout procedures. Methods The National Machine Guarding Program (NMGP) was a translational research initiative implemented in conjunction with two workers' compensation insures. Insurance safety consultants trained in machine guarding used standardize...

Comparison of electrical performances of two n-in-p detectors with different implant type of guard ring by TCAD simulation

Directory of Open Access Journals (Sweden)

Mohammed Mekheldi

Full Text Available This paper presents a preliminary comparative study for two different guard rings structures in the purpose of evaluating their electrical performances. The two structures are based on the n-in-p technology with different implant type of guard rings. I–V characteristics have been simulated using Silvaco/ATLAS software for both structures and compared for various parameters of substrate, guard ring and oxide. Simulation results show that the shape of leakage current is almost the same in all simulations but in terms of breakdown voltage, n-in-p structure with n-type guard rings ensures high voltage stability. Keywords: Breakdown voltage, Guard ring, n-in-p silicon detector, TCAD simulation

The natural history of nest defence in a stingless bee, Tetragonisca angustula (Latreille) (Hymenoptera: Apidae), with two distinct types of entrance guards.

Science.gov (United States)

Grüter, C; Kärcher, M H; Ratnieks, F L W

2011-01-01

The stingless bee Tetragonsica angustula (Latreille) is the only social bee known that has two different types of nest entrance guards. As in other stingless bees and the honey bee one type stands on, in or near the nest entrance. The second type, so far only known in T. angustula, hovers near the nest entrance. In order to gain further understanding of this unique situation we studied guarding behaviour in both types of guards. Using marked bees, we found that individual worker bees guarded for a long time, up to 20 days, relative to their short, average c. 21 day, lifespan. Relatively few, 33%, individually marked guards were seen performing both types of guarding. The others only acted as standing guards. The bees that did perform both types did so over similar periods of their life. Hovering bouts were 57 min long, interrupted by breaks inside the hive of a few minutes (3.3 ± 1.5 min). Standing bouts were slightly longer (74 min) and also interrupted by short breaks (7.82 ± 6.45 min). Human breath, mimicking a vertebrate intruder, caused the guards to retreat into the nest rather than to attack the intruder. Some colonies protected themselves against intruders by closing the entrance during the night (32% and 56% of colonies during two nights). In summary, our results indicate that nest entrance guarding in T. angustula involves division of labour between the two types, in which most guarding individuals only act as standing guards.

Special Staff - Joint Staff - Leadership - The National Guard

Science.gov (United States)

the ARNG Deputy Director of the ARNG Chief of Staff of the ARNG Command Chief Warrant Officer of the Site Maintenance Battle Focused Training Strategy Battle Staff Training Resources News Publications March Today in Guard History Leadership CNGB VCNGB SEA DANG DARNG Joint Staff J-1 J-2 J-3 J-4 J-5 J-6 J

Improved Coast Guard Communications Using Commercial Satellites and WWW Technology

Science.gov (United States)

1997-06-18

Information collection and distribution are essential components of most Coast Guard missions. However, information needs have typically outpaced the ability of the installed communications systems to meet those needs. This mismatch leads to reduced ...

Laser microbeams for the manipulation of plant cells and subcellular structures

International Nuclear Information System (INIS)

Hoffmann, F.

1996-01-01

Laser microsurgery has been used in plants to study physiological, cell biological and genetical questions for over 10 years. More recently, the optical trap became available as an additional tool. Specific areas of research include membrane physiology, motility, transformation and protoplast fusion. Compared to the data reported in animal systems, the contributions of laser microbeam manipulations in plant biology are rather limited. However, with increased awareness of the enormous potential of the technology and better accessibility to less expensive and more user-friendly equipment, the next decade should be more productive. (author)

Coast Guard Acquisitions: Enhanced Oversight of Testing Could Benefit National Security Cutter Program and Future DHS Acquisitions

Science.gov (United States)

2016-02-03

based on our January 2016 report on the NSC’s IOT &E event.2 I will address issues related to (1) the results of the NSC’s IOT &E, (2) the Coast Guard’s...before the testing is complete, meaning that the Coast Guard will be operating 6 NSCs before it has resolved issues from IOT &E and knows the... IOT &E event, the Coast Guard had nearly four years of experience operating the NSCs. The Coast Guard has encountered several issues that require

A metric model of lambda calculus with guarded recursion

DEFF Research Database (Denmark)

Birkedal, Lars; Schwinghammer, Jan; Støvring, Kristian

2010-01-01

We give a model for Nakano’s typed lambda calculus with guarded recursive definitions in a category of metric spaces. By proving a computational adequacy result that relates the interpretation with the operational semantics, we show that the model can be used to reason about contextual equivalence....

77 FR 31183 - Safety Zone, Temporary Change for Recurring Fireworks Display Within the Fifth Coast Guard...

Science.gov (United States)

2012-05-25

...-AA00 Safety Zone, Temporary Change for Recurring Fireworks Display Within the Fifth Coast Guard District, Pamlico River and Tar River; Washington, NC AGENCY: Coast Guard, DHS. ACTION: Temporary final... is intended to restrict vessel traffic in a portion of the Pamlico River and Tar River near...

Receiver sensitivity improvement in spectrally-efficient guard-band twin-SSB-OFDM using an optical IQ modulator

Science.gov (United States)

Chen, Ming; Peng, Miao; Zhou, Hui; Zheng, Zhiwei; Tang, Xionggui; Maivan, Lap

2017-12-01

To further improve receiver sensitivity of spectrally-efficient guard-band direct-detection optical orthogonal frequency-division multiplexing (OFDM) with twin single-side-band (SSB) modulation technique, an optical IQ modulator (IQM) is employed to optimize optical carrier-to-signal power ratio (CSPR). The CSPRs for the guard-band twin-SSB-OFDM signal generated by using dual-drive Mach-Zehnder modulator (DD-MZM) and optical IQM are theoretically analyzed and supported by simulations. The optimal CSPR for the two types of guard-band twin-SSB-OFDM are identified. The simulations exhibit that the error vector magnitude (EVM) performance of the IQM-enabled guard-band twin-SSB-OFDM is improved by more than 4-dB compared to that of the twin-SSB-OFDM enabled by DD-MZM after 80-km single-mode fiber (SMF) transmission. In addition, more than 3-dB and 10 dB receiver sensitivity improvements in terms of received optical power (ROP) and optical signal-to-noise ratio (OSNR) are also achieved, respectively.

Eliminate the Army and Air Force Reserves: Building a Robust National Guard to Meet 21st Century Operational Challenges

Science.gov (United States)

2016-04-04

TITLE AND SUBTITLE ELIMINATE THE ARMY AND AIR FORCE RESERVES: BUILDING A ROBUST NATIONAL GUARD TO MEET 21ST CENTURY OPERATIONAL CHALLENGES 5a...STAFF COLLEGE JOINT ADVANCED WARFIGHTING SCHOOL ELIMINATE THE ARMY AND AIR FORCE RESERVES: BUILDING A ROBUST NATIONAL GUARD TO... ELIMINATE THE ARMY AND AIR FORCE RESERVES: BUILDING A ROBUST NATIONAL GUARD TO MEET 21ST CENTURY OPERATIONAL CHALLENGES by

76 FR 66933 - Privacy Act of 1974; Department of Homeland Security U.S. Coast Guard DHS/USCG-014 Military Pay...

Science.gov (United States)

2011-10-28

... 1974; Department of Homeland Security U.S. Coast Guard DHS/USCG--014 Military Pay and Personnel System... Security U.S. Coast Guard--014 Military Pay and Personnel System of Records.'' This system of records allows the Department of Homeland Security U.S. Coast Guard to collect and maintain records regarding pay...

Effects of ultraviolet radiation on microtubule organisation and morphogenesis in plants

Energy Technology Data Exchange (ETDEWEB)

Staxen, I.

1994-09-01

The involvement of the cytoskeleton in the development of somatic embryos was studied in Larix x eurolepis. Protoplasts were isolated from both somatic embryo-regenerating and non-generating cultures and fractionated on a discontinuous Percoll density gradient. Protoplasts of two cell lines of Larix eurolepis, one with regenerating potential and one lacking this potential, were compared. In contrast to the non-regenerating line were a protoplast-like organisation of the cortical microtubules was maintained, re-organisation of this microtubular network occurred in the regenerable line after only three days of culture, indicating that organised growth was occurring. However, this early organisation of cortical microtubules may not always be a valid marker for regenerable and non-regenerable material. In order to investigate the effect of ultraviolet-B (UV-B, 280-320 nm) radiation on the microtubule cytoskeleton, protoplasts were isolated from leaves of Petunia hybrida and subjected to four different doses of UV-B radiation. The organisation of the microtubules and the progression of the cells through the cell cycle was observed at 0, 24, 48 and 72 h after irradiation. UV-B induced breaks in the cortical microtubules resulting in shorter fragments with increasing amounts of radiation. Also, the division of the protoplasts was delayed. Whole Petunia plants were grown in growth chambers in the presence and absence of UV-B. The plants responded to UV-B with increased rates of CO{sub 2} assimilation, a 60% increase in UV-screening compounds and the changes in the morphology of the leaves that were reflected in a 70-100% increase in leaf area and 20% decrease in leaf thickness. The microtubules of the epidermal cells was not affected by UV-B, nor was the number of epidermal cells (per unit area). The increase in leaf area in the UV-treated plants appeared due to stimulation of cell division in the leaf meristems. 111 refs, 5 figs, 2 tabs.

Effects of over-the-counter jaw-repositioning mouth guards on dynamic balance, flexibility, agility, strength, and power in college-aged male athletes.

Science.gov (United States)

Golem, Devon L; Arent, Shawn M

2015-02-01

Improvements in muscular power and anaerobic performance have resulted from the use of jaw-repositioning mouth guards designed with advanced dental techniques. The high cost of such techniques has dissuaded the widespread use. Recently, more affordable, over-the-counter (OTC) jaw-repositioning mouth guards have become available. The primary objective of this study was to examine the effects of 2 OTC jaw-repositioning mouth guards on muscular power and strength performance in college-aged male athletes. It was hypothesized that similar to previous observations with advanced dentistry-designed mouth guards, OTC jaw-repositioning mouth guards would impart positive effects on muscular power but not have any effect on muscular strength. Secondary objectives of this study included the examination of the effects of 2 OTC jaw-repositioning mouth guards on other variables related to athletic performance. Male collegiate athletes (N = 20) participated in 4 separate testing sessions that consisted of assessment of muscular power, dynamic balance, flexibility, agility, and muscular strength. The 4 conditions, 1 per testing session, were assigned in a randomized order and consisted of a no-mouth guard control (CON), a placebo mouth guard, a self-adapted jaw-repositioning mouth guard (SA), and a custom-fitted jaw-repositioning mouth guard (CF). No significant differences were observed between conditions in muscular power (p = 0.78), dynamic balance (p = 0.99), agility (p = 0.22), or muscular strength (p = 0.47). The CF had significantly lower hip flexion than the CON (p = 0.014) and had significantly greater lumbar spine lateral flexion compared with the SA condition (p = 0.054). However, these flexibility differences lack practical relevance as the effect sizes remain very small (ES = -0.27 and -0.14, respectively). In conclusion, the jaw-repositioning technique used in the design of these OTC mouth guards did not affect performance. It is important to note that negative

Adaptive guard channel allocation scheme with buffer for mobile ...

African Journals Online (AJOL)

The devastating effect congestion has on the quality of service delivery and overall network performance demands an utmost attention. This certainly calls for taking some expedient measures to deal with congestion so as to salvage the network from total collapse. In this paper, an adaptive guard channel allocation scheme ...

Mutational analysis of the RNA-binding domain of the Prunus necrotic ringspot virus (PNRSV) movement protein reveals its requirement for cell-to-cell movement.

Science.gov (United States)

Carmen Herranz, Ma; Sanchez-Navarro, Jesús-Angel; Saurí, Ana; Mingarro, Ismael; Pallás, Vicente

2005-08-15

The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is required for cell-to-cell movement. MP subcellular localization studies using a GFP fusion protein revealed highly punctate structures between neighboring cells, believed to represent plasmodesmata. Deletion of the RNA-binding domain (RBD) of PNRSV MP abolishes the cell-to-cell movement. A mutational analysis on this RBD was performed in order to identify in vivo the features that govern viral transport. Loss of positive charges prevented the cell-to-cell movement even though all mutants showed a similar accumulation level in protoplasts to those observed with the wild-type (wt) MP. Synthetic peptides representing the mutants and wild-type RBDs were used to study RNA-binding affinities by EMSA assays being approximately 20-fold lower in the mutants. Circular dichroism analyses revealed that the secondary structure of the peptides was not significantly affected by mutations. The involvement of the affinity changes between the viral RNA and the MP in the viral cell-to-cell movement is discussed.

Mutational analysis of the RNA-binding domain of the Prunus necrotic ringspot virus (PNRSV) movement protein reveals its requirement for cell-to-cell movement

International Nuclear Information System (INIS)

Carmen Herranz, Ma; Sanchez-Navarro, Jesus-Angel; Sauri, Ana; Mingarro, Ismael; Pallas, Vicente

2005-01-01

The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is required for cell-to-cell movement. MP subcellular localization studies using a GFP fusion protein revealed highly punctate structures between neighboring cells, believed to represent plasmodesmata. Deletion of the RNA-binding domain (RBD) of PNRSV MP abolishes the cell-to-cell movement. A mutational analysis on this RBD was performed in order to identify in vivo the features that govern viral transport. Loss of positive charges prevented the cell-to-cell movement even though all mutants showed a similar accumulation level in protoplasts to those observed with the wild-type (wt) MP. Synthetic peptides representing the mutants and wild-type RBDs were used to study RNA-binding affinities by EMSA assays being approximately 20-fold lower in the mutants. Circular dichroism analyses revealed that the secondary structure of the peptides was not significantly affected by mutations. The involvement of the affinity changes between the viral RNA and the MP in the viral cell-to-cell movement is discussed

75 FR 79956 - Protection for Whistleblowers in the Coast Guard

Science.gov (United States)

2010-12-21

... constitutes evidence of sexual harassment and discrimination, among other subjects. Additionally, changes to... (including sexual harassment or discrimination), gross mismanagement, a gross waste of funds or other... Force, Marine Corps, and Coast Guard''). Section 1034 covers allegations and disclosures of sexual...

Comparison of electrical performances of two n-in-p detectors with different implant type of guard ring by TCAD simulation

Science.gov (United States)

Mekheldi, Mohammed; Oussalah, Slimane; Lounis, Abdenour; Brihi, Nourredine

This paper presents a preliminary comparative study for two different guard rings structures in the purpose of evaluating their electrical performances. The two structures are based on the n-in-p technology with different implant type of guard rings. I-V characteristics have been simulated using Silvaco/ATLAS software for both structures and compared for various parameters of substrate, guard ring and oxide. Simulation results show that the shape of leakage current is almost the same in all simulations but in terms of breakdown voltage, n-in-p structure with n-type guard rings ensures high voltage stability.

Army Aviation in the National Guard: Assessing for Efficiency and Effectiveness

National Research Council Canada - National Science Library

Gattis, Karen D

2009-01-01

...). This SRP examines Army National Guard Aviation to determine if the force structure can remain balanced as an effective and efficient operational force while maintaining the OPTEMPO in support of GWOT...

TrustGuard: A Containment Architecture with Verified Output

Science.gov (United States)

2017-01-01

that the TrustGuard system has minimal performance decline, despite restrictions such as high communication latency and limited available bandwidth...design are the availability of high bandwidth and low delays between the host and the monitoring chip. 3-D integration provides an alternate way of...TRUSTGUARD: A CONTAINMENT ARCHITECTURE WITH VERIFIED OUTPUT SOUMYADEEP GHOSH A DISSERTATION PRESENTED TO THE FACULTY OF PRINCETON UNIVERSITY IN

US Coast Guard national spill response resource inventory

International Nuclear Information System (INIS)

Giraitis, D.M.

1993-01-01

The Oil Pollution Act of 1990 (OPA 90) mandated the establishment of a National Response Unit, now renamed the National Strike Force Coordination Center (NSFCC). Among the duties OPA 90 assigned to this new Coast Guard unit was to compile and maintain a comprehensive list of spill removal resources, personnel, and equipment that is available to Federal and State agencies and to the public. The Coast Guard's Research and Development Center has been developing this project, the Response Resource Inventory (RRI), for one year. The RRI is expected to be operational, with resource data from industry in the data base, by the time of the International Oil Spill Conference in March 1993. The RRI will contain data on skimmers, specialized oil recovery vessels, oil/water separators, dispersants and delivery systems, etc. Previous attempts to maintain a national data base of these response resources, including an earlier Coast Guard system called the Spill Cleanup Equipment Inventory System (SKIM), fell into disuse for a number of reasons. Inaccuracies caused by inadvertent double counting of equipment and outdated information coupled with difficulties in accessing the information were common shortfalls of early systems. During the development of the RRI, user-group meetings were held to determine system requirements and study the failures of previous systems. Methods of obtaining and entering data were a major concern during RRI development. A data collection system that supplies the resource holder with a DOS-formatted diskette containing a collection program will be used. The program will make extensive use of pick lists to minimize the data entry burden on the resource holder and maintain standard entries. When the program is completed, the diskette will be mailed to the NSFCC, where the data will be transferred quickly to the RRI data base. Easy access to the data for the public and industry will be afforded mainly through a computer bulletin board

Meeting the Challenge - Developing Leaders for Army National Guard Combat Units

National Research Council Canada - National Science Library

Keolanui, Stan

2000-01-01

...). Capable and qualified leaders are a necessity in complex organizations like the eSB. These leaders will insure that Guard combat units meet all readiness and deployment standards necessary for future combat...

33 CFR 165.117 - Regulated Navigation Areas, Safety and Security Zones: Deepwater Ports, First Coast Guard District.

Science.gov (United States)

2010-07-01

..., Safety and Security Zones: Deepwater Ports, First Coast Guard District. 165.117 Section 165.117... Limited Access Areas First Coast Guard District § 165.117 Regulated Navigation Areas, Safety and Security... section are designated as regulated navigation areas. (2) Safety and security zones. All waters within a...

33 CFR 165.168 - Safety Zones; Coast Guard Captain of the Port New York Fireworks Displays.

Science.gov (United States)

2010-07-01

... of the Port New York Fireworks Displays. 165.168 Section 165.168 Navigation and Navigable Waters... Guard District § 165.168 Safety Zones; Coast Guard Captain of the Port New York Fireworks Displays. (a... Upper New York Bay within a 360-yard radius of the fireworks barge in approximate position 40°41′16.5″ N...

The “Suicide Guard Rail”: a minimal structural intervention in hospitals reduces suicide jumps

OpenAIRE

Mohl, Andreas; Stulz, Niklaus; Martin, Andrea; Eigenmann, Franz; Hepp, Urs; Hüsler, Jürg; Beer, Jürg H

2012-01-01

Abstract Background Jumping from heights is a readily available and lethal method of suicide. This study examined the effectiveness of a minimal structural intervention in preventing suicide jumps at a Swiss general teaching hospital. Following a series of suicide jumps out of the hospital’s windows, a metal guard rail was installed at each window of the high-rise building. Results In the 114 months prior to the installation of the metal guard rail, 10 suicides by jumping out of the hospital’...

Different genome maintenance strategies in human and tobacco cells.

Science.gov (United States)

Pelczar, Pawel; Kalck, Véronique; Kovalchuk, Igor

2003-08-22

In this work, genome maintenance strategies of organisms belonging to different kingdoms (animals versus plants) but of similar genome size were investigated using a novel, universal double-strand break (DSB) repair assay. Different plasmids linearised with KpnI, Acc65I or EcoRV yielding either 3' or 5' protruding or blunt DNA termini, respectively, were transfected into HeLa cells and Nicotiana plumbaginifolia protoplasts and assayed for the efficiency and fidelity of DSB repair. We show that the mechanism of break sealing is similar but that drastic differences are seen in the fidelity of repair: in HeLa cells, 50-55% DSBs were repaired precisely, compared to as little as 15-30% in tobacco cells. Moreover, the DSB repair in plants resulted in 30-40% longer deletions and significantly shorter insertions. Combined, these led to more than twofold larger net DNA loss in tobacco cells. Our observations point to possible differences in the strategies of DSB repair and genome maintenance in plants and animals.

Cdc45 (cell division cycle protein 45) guards the gate of the Eukaryote Replisome helicase stabilizing leading strand engagement

Science.gov (United States)

Petojevic, Tatjana; Pesavento, James J.; Costa, Alessandro; Liang, Jingdan; Wang, Zhijun; Berger, James M.; Botchan, Michael R.

2015-01-01

DNA replication licensing is now understood to be the pathway that leads to the assembly of double hexamers of minichromosome maintenance (Mcm2–7) at origin sites. Cell division control protein 45 (Cdc45) and GINS proteins activate the latent Mcm2–7 helicase by inducing allosteric changes through binding, forming a Cdc45/Mcm2-7/GINS (CMG) complex that is competent to unwind duplex DNA. The CMG has an active gate between subunits Mcm2 and Mcm5 that opens and closes in response to nucleotide binding. The consequences of inappropriate Mcm2/5 gate actuation and the role of a side channel formed between GINS/Cdc45 and the outer edge of the Mcm2–7 ring for unwinding have remained unexplored. Here we uncover a novel function for Cdc45. Cross-linking studies trace the path of the DNA with the CMG complex at a fork junction between duplex and single strands with the bound CMG in an open or closed gate conformation. In the closed state, the lagging strand does not pass through the side channel, but in the open state, the leading strand surprisingly interacts with Cdc45. Mutations in the recombination protein J fold of Cdc45 that ablate this interaction diminish helicase activity. These data indicate that Cdc45 serves as a shield to guard against occasional slippage of the leading strand from the core channel. PMID:25561522

33 CFR 165.161 - Safety zones: Coast Guard Captain of the Port New York annual fireworks displays.

Science.gov (United States)

2010-07-01

... of the Port New York annual fireworks displays. 165.161 Section 165.161 Navigation and Navigable... Coast Guard District § 165.161 Safety zones: Coast Guard Captain of the Port New York annual fireworks displays. (a) Safety zones. The following areas are designated safety zones: (1) Bar Beach fireworks...

National Machine Guarding Program: Part 1. Machine safeguarding practices in small metal fabrication businesses.

Science.gov (United States)

Parker, David L; Yamin, Samuel C; Brosseau, Lisa M; Xi, Min; Gordon, Robert; Most, Ivan G; Stanley, Rodney

2015-11-01

Metal fabrication workers experience high rates of traumatic occupational injuries. Machine operators in particular face high risks, often stemming from the absence or improper use of machine safeguarding or the failure to implement lockout procedures. The National Machine Guarding Program (NMGP) was a translational research initiative implemented in conjunction with two workers' compensation insures. Insurance safety consultants trained in machine guarding used standardized checklists to conduct a baseline inspection of machine-related hazards in 221 business. Safeguards at the point of operation were missing or inadequate on 33% of machines. Safeguards for other mechanical hazards were missing on 28% of machines. Older machines were both widely used and less likely than newer machines to be properly guarded. Lockout/tagout procedures were posted at only 9% of machine workstations. The NMGP demonstrates a need for improvement in many aspects of machine safety and lockout in small metal fabrication businesses. © 2015 The Authors. American Journal of Industrial Medicine published by Wiley Periodicals, Inc.

Inter-Symbol Guard Time for Synchronizing Optical PPM

Science.gov (United States)

Far, William; Gin, Jonathan; Srinivasan, Meera; Quirk, Kevin

2009-01-01

An inter-symbol guard time has been proposed as a means of synchronizing the symbol and slot clocks of an optical pulse-position modulation (PPM) receiver with the symbol and slot periods of an incoming optical PPM signal.The proposal is applicable to the low-flux case in which the receiver photodetector operates in a photon-counting mode and the count can include contributions from incidental light sources and dark current.

Risk, Resiliency and Coping in National Guard Families

Science.gov (United States)

2015-10-01

deployment, to National Guard in second deployment to civilian employment. Couple said they were careful with their finances and worked hard to save money ...were very helpful in providing rent money when the couple was struggling and their children were able to get healthcare through a government subsidized...is examining risk and resilience factors for various family types ( couples , families with children, single NG with and without parental support

Michigan field artillery's 'Blackjacks' training in Latvia > National Guard

Science.gov (United States)

Charles Smith, 1st Battalion, 119th Field Artillery, Michigan Army National Guard, set up the M67 GLPS for 24 years. 1st. Lt. Brice Masterson, and Sgt. 1st Class Charles Smith, 1st Battalion, 119th Field , Lithuania and the United States will participate in the exercise. The exercise takes place at Adazi Training

Suicide in the Army National Guard: An Empirical Inquiry

Science.gov (United States)

Griffith, James

2012-01-01

Since 2004, suicides in the U.S. military have risen, most notably in the Army National Guard (ARNG). Data used in this study were obtained for suicides occurring from 2007 to 2010 and for a random sample of nonsuicides from the general ARNG population. Of the military-related variables considered, a few showed relationships to suicide. Rather,…

Utilization of glucose and UDPG by supprotoplasts of cotton fiber cells

International Nuclear Information System (INIS)

Gould, J.H.; Dugger, W.M.

1984-01-01

The authors have developed a subprotoplast system for cotton fiber cells isolated after initiation of secondary wall and cellulose synthesis. In the absence of a cell-free system for cellulose synthesis, protoplasts and subprotoplasts offer an opportunity to study cellulose synthesis as well as precursor utilization. In these systems, however, the incorporation of precursor is confused by an unknown mode of uptake from the culture medium. These studies were undertaken to clarify the uptake question. Results could corroborate a model of UDP-glucose utilization at the plasma membrane surface or uptake of an intact molecule. The cotton fiber subprotoplast system appears to synthesize a product characteristic of cellulose in enough quantity for further characterization, and may prove to be useful in studying some aspects of cellulose synthesis

The Cell Wall Protein Ecm33 of Candida albicans is Involved in Chronological Life Span, Morphogenesis, Cell Wall Regeneration, Stress Tolerance, and Host-Cell Interaction.

Science.gov (United States)

Gil-Bona, Ana; Reales-Calderon, Jose A; Parra-Giraldo, Claudia M; Martinez-Lopez, Raquel; Monteoliva, Lucia; Gil, Concha

2016-01-01

Ecm33 is a glycosylphosphatidylinositol-anchored protein in the human pathogen Candida albicans. This protein is known to be involved in fungal cell wall integrity (CWI) and is also critical for normal virulence in the mouse model of hematogenously disseminated candidiasis, but its function remains unknown. In this work, several phenotypic analyses of the C. albicans ecm33/ecm33 mutant (RML2U) were performed. We observed that RML2U displays the inability of protoplast to regenerate the cell wall, activation of the CWI pathway, hypersensitivity to temperature, osmotic and oxidative stresses and a shortened chronological lifespan. During the exponential and stationary culture phases, nuclear and actin staining revealed the possible arrest of the cell cycle in RML2U cells. Interestingly, a "veil growth," never previously described in C. albicans, was serendipitously observed under static stationary cells. The cells that formed this structure were also observed in cornmeal liquid cultures. These cells are giant, round cells, without DNA, and contain large vacuoles, similar to autophagic cells observed in other fungi. Furthermore, RML2U was phagocytozed more than the wild-type strain by macrophages at earlier time points, but the damage caused to the mouse cells was less than with the wild-type strain. Additionally, the percentage of RML2U apoptotic cells after interaction with macrophages was fewer than in the wild-type strain.

77 FR 37318 - Eighth Coast Guard District Annual Safety Zones; Sound of Independence; Santa Rosa Sound; Fort...

Science.gov (United States)

2012-06-21

...-AA00 Eighth Coast Guard District Annual Safety Zones; Sound of Independence; Santa Rosa Sound; Fort... Coast Guard will enforce a Safety Zone for the Sound of Independence event in the Santa Rosa Sound, Fort... during the Sound of Independence. During the enforcement period, entry into, transiting or anchoring in...

The Defense Message System and the U.S. Coast Guard

Science.gov (United States)

1992-06-01

these mail services, the Internet also provides a File Transfer Protocol (FTP) and remote login between host computers (TELNET) capabilities. 17 [Ref...the Joint Maritime Intelligence Element (JMIE), Zincdust, and Emerald . [Ref. 27] 4. Secure Data Network The Coast Guard’s Secure Data Network (SDN

The Coast Guard Intelligence Program Enters the Intelligence Community. A Case Study of Congressional Influence on Intelligence Community Evolution

National Research Council Canada - National Science Library

Wirth, Kevin E

2007-01-01

.... Although the United States Coast Guard has utilized intelligence capabilities since the service's inception in 1790, the Coast Guard was not included as a formal member of the Intelligence Community until December 2002. Mr...

Coast Guard: Progress Being Made on Deepwater Project, but Risks Remain

National Research Council Canada - National Science Library

2001-01-01

The Coast Guard is in the final stages of planning the largest procurement project in its history the modernization and/or replacement of over 90 cutters and 200 aircraft used for missions beyond 50 miles from shore...

Stomatal Responses to Flooding of the Intercellular Air Spaces Suggest a Vapor-Phase Signal Between the Mesophyll and the Guard Cells1[OA

Science.gov (United States)

Sibbernsen, Erik; Mott, Keith A.

2010-01-01

Flooding the intercellular air spaces of leaves with water was shown to cause rapid closure of stomata in Tradescantia pallida, Lactuca serriola, Helianthus annuus, and Oenothera caespitosa. The response occurred when water was injected into the intercellular spaces, vacuum infiltrated into the intercellular spaces, or forced into the intercellular spaces by pressurizing the xylem. Injecting 50 mm KCl or silicone oil into the intercellular spaces also caused stomata to close, but the response was slower than with distilled water. Epidermis-mesophyll grafts for T. pallida were created by placing the epidermis of one leaf onto the exposed mesophyll of another leaf. Stomata in these grafts opened under light but closed rapidly when water was allowed to wick between epidermis and the mesophyll. When epidermis-mesophyll grafts were constructed with a thin hydrophobic filter between the mesophyll and epidermis stomata responded normally to light and CO2. These data, when taken together, suggest that the effect of water on stomata is caused partly by dilution of K+ in the guard cell and partly by the existence of a vapor-phase signal that originates in the mesophyll and causes stomata to open in the light. PMID:20472750

Organization of the cytoplasmic reticulum in the central vacuole of parenchyma cells in Allium cepa L.

Directory of Open Access Journals (Sweden)

Tomasz J. Wodzicki

2015-01-01

Full Text Available An elaborate and complex cytoplasmic reticulum composed of fine filaments and lamellae ranging from 0.1 to 4 microns in size is revealed by viewing the central vacuole of onion bulb parenchyma cells with the scanning election microscope. The larger cytoplasmic strands, visible with the light microscope, are composed of numerous smaller filaments (some tubular which might explain the observed bidirectional movement of particles in these larger strands. The finely divided cytoplasmic network of filaments is continuous with the parietal cytoplasm inclosing the vacuolar sap. In these highly vacuolated cells the mass of the protoplast is in the form of an intravacuolar reticulum immersed in the cell sap. The probable significance of the vacuolar sap in relation to physiological processes of the cell is discussed.

The Coast Guard’s Critical Role as an Armed Service

Science.gov (United States)

2012-03-22

security authorities. Although the Revenue Cutter Service under the Department of the Treasury was primarily responsible for the enforcement of taxation ...maritime exercises with Brunei, Thailand, Indonesia, Singapore, and Malaysia .15 Coast Guard cutters and Law Enforcement Detachments (LEDETs

Single channel and WDM transmission of 28 Gbaud zero-guard-interval CO-OFDM.

Science.gov (United States)

Zhuge, Qunbi; Morsy-Osman, Mohamed; Mousa-Pasandi, Mohammad E; Xu, Xian; Chagnon, Mathieu; El-Sahn, Ziad A; Chen, Chen; Plant, David V

2012-12-10

We report on the experimental demonstration of single channel 28 Gbaud QPSK and 16-QAM zero-guard-interval (ZGI) CO-OFDM transmission with only 1.34% overhead for OFDM processing. The achieved transmission distance is 5120 km for QPSK assuming a 7% forward error correction (FEC) overhead, and 1280 km for 16-QAM assuming a 20% FEC overhead. We also demonstrate the improved tolerance of ZGI CO-OFDM to residual inter-symbol interference compared to reduced-guard-interval (RGI) CO-OFDM. In addition, we report an 8-channel wavelength-division multiplexing (WDM) transmission of 28 Gbaud QPSK ZGI CO-OFDM signals over 4160 km.

Photosynthesis in Flaveria brownii, a C(4)-Like Species: Leaf Anatomy, Characteristics of CO(2) Exchange, Compartmentation of Photosynthetic Enzymes, and Metabolism of CO(2).

Science.gov (United States)

Cheng, S H; Moore, B D; Edwards, G E; Ku, M S

1988-08-01

Light microscopic examination of leaf cross-sections showed that Flaveria brownii A. M. Powell exhibits Kranz anatomy, in which distinct, chloroplast-containing bundle sheath cells are surrounded by two types of mesophyll cells. Smaller mesophyll cells containing many chloroplasts are arranged around the bundle sheath cells. Larger, spongy mesophyll cells, having fewer chloroplasts, are located between the smaller mesophyll cells and the epidermis. F. brownii has very low CO(2) compensation points at different O(2) levels, which is typical of C(4) plants, yet it does show about 4% inhibition of net photosynthesis by 21% O(2) at 30 degrees C. Protoplasts of the three photosynthetic leaf cell types were isolated according to relative differences in their buoyant densities. On a chlorophyll basis, the activities of phosphoenolpyruvate carboxylase and pyruvate, Pi dikinase (carboxylation phase of C(4) pathway) were highest in the larger mesophyll protoplasts, intermediate in the smaller mesophyll protoplasts, and lowest, but still present, in the bundle sheath protoplasts. In contrast, activities of ribulose 1,5-bisphosphate carboxylase, other C(3) cycle enzymes, and NADP-malic enzyme showed a reverse gradation, although there were significant activities of these enzymes in mesophyll cells. As indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the banding pattern of certain polypeptides of the total soluble proteins from the three cell types also supported the distribution pattern obtained by activity assays of these enzymes. Analysis of initial (14)C products in whole leaves and extrapolation of pulse-labeling curves to zero time indicated that about 80% of the CO(2) is fixed into C(4) acids (malate and aspartate), whereas about 20% of the CO(2) directly enters the C(3) cycle. This is consistent with the high activity of enzymes for CO(2) fixation by the C(4) pathway and the substantial activity of enzymes of the C(3) cycle in the mesophyll cells

STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

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T. VINTILĂ

2007-05-01

Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmid vectors (pLC1 and pNC61, using electroporation technique, protoplast transformation and bivalent cations (CaCl2 mediated transformation. In the case of transformation by electroporation of Bacillus licheniformis B40, the highest number of transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2 milliseconds. Using this transformation technique we have obtained six kanamycin resistant transformants. The frequency of Bacillus licheniformis B40 protoplasts transformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF = 10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts, six kanamycin resistant transformants were obtained. The pNC61 plasmid, which confers trimethoprim resistance, does not integrate in receiver cells by protoplast transformation. The direct genetic transformation in the presence of bivalent cations (CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a low transformation frequency. Using this technique, we have obtained three trimethoprim resistant colonies and four kanamycin resistant colonies. The chemical way of transformation is the only technique, which realizes the integration of pNC61 in B. licheniformis B40 cells.

76 FR 53329 - Eleventh Coast Guard District Annual Marine Events

Science.gov (United States)

2011-08-26

.... SUMMARY: The Coast Guard is updating and consolidating the list of marine events occurring annually within... not currently reflect changes in actual dates and other required information. The effects of these... location that offers a better vantage point to monitor the event. The location of the PATCOM may also be...

46 CFR 159.005-7 - Preapproval review: Coast Guard action.

Science.gov (United States)

2010-10-01

... Section 159.005-7 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT... may be conducted. (2) If the Commandant determines from the application for approval that the... under § 159.005-13 can be taken. (c) An item of equipment or material that does not meet all of the...

33 CFR 173.85 - Fees levied by the Coast Guard.

Science.gov (United States)

2010-07-01

.... 173.85 Section 173.85 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) BOATING SAFETY VESSEL NUMBERING AND CASUALTY AND ACCIDENT REPORTING Issue of Certificate of.... (b) Fees are payable by check or money-order made payable to the “U.S. Coast Guard”; by major credit...

Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: cytological studies.

Science.gov (United States)

Rakkhumkaew, Numfon; Shibatani, Shigeo; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

2013-04-01

Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured-cells (BY-2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY-2 cells harboring a vSPO-cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N-terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA. Copyright © 2012 Wiley Periodicals, Inc.

Assessment of a postdeployment Yellow Ribbon Reintegration Program for National Guard members and supporters.

Science.gov (United States)

Scherrer, Jeffrey F; Widner, Greg; Shroff, Manan; Matthieu, Monica; Balan, Sundari; van den Berk-Clark, Carissa; Price, Rumi K

2014-11-01

The Yellow Ribbon Reintegration Program (YRRP) was created to meet the needs of National Guard members and their families throughout the deployment cycle. This study examined the perceived utility of the YRRP's delivery of information and assistance during the postdeployment reintegration period by National Guard members and accompanying supporters who were mostly spouses. Over 22 months, from 10 YRRP events, 683 service members and 411 supporters completed questionnaires immediately after the YRRP. We analyzed questions on information and avenues for help, timeliness and concerns related to education, employment, legal, family, and health. Service members and supporters most often endorsed information delivery on education being met (76.8% and 78.2%, respectively) and were least likely to endorse legal information delivery (63.5% and 60%, respectively). Significantly more supporters than service members (p family. Results suggest the YRRP fills gaps in supporter knowledge and provides needed information and resources to most National Guard families 2 to 4 months after a deployment. Reprint & Copyright © 2014 Association of Military Surgeons of the U.S.

National machine guarding program: Part 1. Machine safeguarding practices in small metal fabrication businesses

Science.gov (United States)

Yamin, Samuel C.; Brosseau, Lisa M.; Xi, Min; Gordon, Robert; Most, Ivan G.; Stanley, Rodney

2015-01-01

Background Metal fabrication workers experience high rates of traumatic occupational injuries. Machine operators in particular face high risks, often stemming from the absence or improper use of machine safeguarding or the failure to implement lockout procedures. Methods The National Machine Guarding Program (NMGP) was a translational research initiative implemented in conjunction with two workers' compensation insures. Insurance safety consultants trained in machine guarding used standardized checklists to conduct a baseline inspection of machine‐related hazards in 221 business. Results Safeguards at the point of operation were missing or inadequate on 33% of machines. Safeguards for other mechanical hazards were missing on 28% of machines. Older machines were both widely used and less likely than newer machines to be properly guarded. Lockout/tagout procedures were posted at only 9% of machine workstations. Conclusions The NMGP demonstrates a need for improvement in many aspects of machine safety and lockout in small metal fabrication businesses. Am. J. Ind. Med. 58:1174–1183, 2015. © 2015 The Authors. American Journal of Industrial Medicine published by Wiley Periodicals, Inc. PMID:26332060

Environmental Service and Outdoor Adventure as a Context for Positive Youth Development: An Evaluation of the Crow River Trail Guards Program

Directory of Open Access Journals (Sweden)

Julie Ernst

2013-06-01

Full Text Available Trail Guards, a community-based organization in Minnesota, offers youth the opportunity to participate in park/trail maintenance and enhancement projects. Through these environmental service projects, Trail Guards seeks to foster the following developmental outcomes in youth participants: self-awareness of skills and strengths; self-worth; personal and social self-efficacy; sense of belonging and acceptance; team work and cooperation skills; and a sense of community responsibility. Trail Guards ultimately aims for youth to transfer these skills and socially appropriate behaviors to settings and activities beyond Trail Guards and to participate in the community in other positive ways. A program evaluation indicated Trail Guards seems to be achieving these youth development outcomes, and that the success of the program may be attributed to the program leader serving as a positive adult role model and providing a safe and caring environment, as well as to community involvement. Implications are discussed.

Apparent inhibition of β-fructosidase secretion by tunicamycin may be explained by breakdown of the unglycosylated protein during secretion

International Nuclear Information System (INIS)

Faye, L.; Chrispeels, M.J.

1989-01-01

Suspension-cultured carrot (Daucus carota) cells synthesize and secrete β-fructosidase, a glycoprotein with asparagine-linked glycans. Treatment of the cells with tunicamycin completely inhibits the apparent secretion of β-fructosidase as measured by the accumulation of the 35 S-labelled protein in the cell wall or the culture medium. In the past, such a result has been interpreted as an inhibition of secretion by tunicamycin, but we suggest another explanation based on the following results. In the presence of tunicamycin, unglycosylated β-fructosidase is synthesized and is associated with an endoplasmic-reticulum-rich microsomal fraction. Pulse-chase experiments show that the unglycosylated β-fructosidase does not remain in the cells and appears to be secreted in the same way as glycosylated β-fructosidase; however, no radioactive, unglycosylated β-fructosidase accumulates extracellularly (cell wall or medium). Protoplasts obtained from carrot cells secrete β-fructosidase protein and activity, and treatment of the protoplasts with tunicamycin results in the synthesis of unglycosylated β-fructosidase. In the presence of tunicamycin, there is no accumulation of β-fructosidase activity or unglycosylated β-fructosidase polypeptide in the protoplast incubation medium. These results are consistent with the interpretation that the glycans of β-fructosidase are necessary for its stability, and that in these suspension-cultured cells, the unglycosylated enzyme is degraded during the last stage(s) of secretion, or immediately after its arrival in the wall

Assessment of a Post-deployment Yellow Ribbon Reintegration Program for National Guard Members and Supporters

Science.gov (United States)

Scherrer, Jeffrey F.; Widner, Greg; Shroff, Manan; Matthieu, Monica; Balan, Sundari; van den Berk-Clark, Carissa; Price, Rumi Kato

2014-01-01

The Yellow Ribbon Reintegration Program (YRRP) was created to meet the needs of National Guard members and their families throughout the deployment cycle. This study examined the perceived utility of the YRRP’s delivery of information and assistance during the post-deployment reintegration period by National Guard members and accompanying supporters who were mostly spouses. Over 22 months, from 10 YRRP events, 683 service members and 411 supporters completed questionnaires immediately after the YRRP. We analyzed questions on information and help provision, timeliness and concerns related to education, employment, legal, family, and health. Service members and supporters most often endorsed education needs being met (76.8% and 78.2% respectively) and were least likely to endorse legal needs being met (63.5% and 60% respectively). Significantly more supporters than service members (p family. Results suggest the YRRP fills gaps in supporter knowledge and provides needed information and resources to most National Guard families 2-4 months after a deployment. PMID:25373071

Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. [Annual report], May 16, 1993--January 29, 1994

Energy Technology Data Exchange (ETDEWEB)

Steponkus, P.L.

1994-06-01

Our aim is to provide a mechanistic understanding of the cellular and molecular aspects of freezing injury and cold acclimation from a perspective of the structural and functional integrity of the plasma membrane-the primary site of freezing injury in winter cereals. We established that destabilization of the plasma membrane of winter rye, the most freezing-tolerant winter cereal, can result from several different lesions: expansion induced lysis, lamellar-to-hexagonal II phase transitions, and the fracture-jump lesion. The occurrence and incidence of these various lesions, depends on the freeze/thaw protocol and the stage of cold acclimation. In non-acclimated leaves and protoplasts, expansion-induced lysis is the predominant lesion at temperatures between {minus}2 and {minus}5{degree}C, whereas freeze-induced formation of the H{sub II} phase is the predominant lesion at temperatures below {minus}10{degree}C. We investigated whether the difference in freezing tolerance and the threshold temperatures at which the lesions occur in rye and oat are a consequence of differences in the lipid composition of the plasma membrane. There are substantial differences between rye and oat cell membranes both before and after cold acclimation. The plasma membrane of oat contains greater proportions of acylated sterylglucosides and cerebrosides than that of rye, and there is little change in these two lipid classes during cold acclimation. The lyotropic phase behavior of lipid mixtures that resemble the plasma membrane of rye and oat was studied. The differences in lipid composition of rye and oat are of mechanistic significance because of their influence on the hydration characteristics of the plasma membrane, the propensity for dehydration-induced lipid-lipid demixing, and the intrinsic curvature of the lipid monolayers. These studies suggest that strategies for improving the freezing tolerance of winter cereals should include approaches to modify membrane lipid composition.

78 FR 53675 - Eighth Coast Guard District Annual Safety Zones; Boomsday Festival; Tennessee River 646.0-649.0...

Science.gov (United States)

2013-08-30

...-AA00 Eighth Coast Guard District Annual Safety Zones; Boomsday Festival; Tennessee River 646.0-649.0... Guard will enforce a Safety Zone for the Boomsday Festival Fireworks on the Tennessee River 646.0-649.0... Festival Fireworks. During the enforcement period, entry into, transiting or anchoring in the Safety Zone...

Postdeployment reintegration experiences of female soldiers from national guard and reserve units in the United States.

Science.gov (United States)

Kelly, Patricia J; Berkel, LaVerne A; Nilsson, Johanna E

2014-01-01

Women are an integral part of Reserve and National Guard units and active duty armed forces of the United States. Deployment to conflict and war zones is a difficult experience for both soldiers and their families. On return from deployment, all soldiers face the challenge of reintegration into family life and society, but those from the National Guard and Reserve units face the additional challenge of reintegration in relative isolation from other soldiers. There is limited research about the reintegration experiences of women and the functioning of the families during reintegration following deployment. The goal was to document postdeployment family reintegration experiences of women in the National Guard. Semistructured interviews were conducted with 42 female members of Midwestern National Guard units. Directed content analysis was used to identify categories of experiences related to women's family reintegration. Five categories of postdeployment experience for female soldiers and their families were identified: Life Is More Complex, Loss of Military Role, Deployment Changes You, Reestablishing Partner Connections, and Being Mom Again. The categories reflected individual and family issues, and both need to be considered when soldiers and their families seek care. Additional research is needed to fully understand the specific impact of gender on women's reintegration.

Findings From the National Machine Guarding Program–A Small Business Intervention

Science.gov (United States)

Parker, David L.; Yamin, Samuel C.; Xi, Min; Brosseau, Lisa M.; Gordon, Robert; Most, Ivan G.; Stanley, Rodney

2016-01-01

Objectives: The purpose of this nationwide intervention was to improve machine safety in small metal fabrication businesses (3 to 150 employees). The failure to implement machine safety programs related to guarding and lockout/tagout (LOTO) are frequent causes of Occupational Safety and Health Administration (OSHA) citations and may result in serious traumatic injury. Methods: Insurance safety consultants conducted a standardized evaluation of machine guarding, safety programs, and LOTO. Businesses received a baseline evaluation, two intervention visits, and a 12-month follow-up evaluation. Results: The intervention was completed by 160 businesses. Adding a safety committee was associated with a 10% point increase in business-level machine scores (P < 0.0001) and a 33% point increase in LOTO program scores (P < 0.0001). Conclusions: Insurance safety consultants proved effective at disseminating a machine safety and LOTO intervention via management-employee safety committees. PMID:27466709

Histochemical evidence for the relationship between peel damage and the accumulation of phenolic compounds in gamma-irradiated citrus fruit

International Nuclear Information System (INIS)

Riov, J.

1975-01-01

The first symptoms of radiation damage to citrus fruit were noted 24 to 48 hr after irradiation with 240 krad of gamma rays. At about the same time, radiation-damaged cells were observed in peel cross sections in the flavedo, the outer colored peel layers. The damaged cells were deformed and their protoplasts stained much darker with haematoxylin-safranin than protoplasts of intact cells. The cytoplasm in damaged cells either thickened at one side of the cell, sometimes filling up most of the cell space, or contracted into a small mass. Using several histochemical reagents, a considerable accumulation of phenolic compounds was found to occur in the damaged cells shortly after irradiation. No accumulation of phenolic compounds was observed in intact cells of irradiated fruit. It is suggested that the phenolic compounds which accumulate in flavedo cells following irradiation cause cell death and consequent peel necrosis (pitting). (author)

Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Summary progress report, May 16, 1987--June 1, 1991

Energy Technology Data Exchange (ETDEWEB)

Steponkus, P.L.

1991-12-31

This project focuses on lesions in the plasma membrane of protoplasts that occur during freezing to temperatures below {minus}5{degrees} which result in changes in the semipermeablity of the plasma membrane. This injury, referred to as loss of osmotic responsiveness, is associated with the formation of large, aparticulate domains in the plasma membrane, aparticulate lamellae subtending the plasma membrane, and lamellar-to-hexagonal{sub II} phase transitions in the plasma membrane and subtending lamellar. The goals of this project are to provide a mechanistic understanding of the mechanism by which freeze-induced dehydration effects the formation of aparticulate domains and lamellar-to-hexagonal{sub II} phase transitions and to determine the mechanisms by which cold acclimation and cryoprotectants preclude or diminish these ultrastructural changes. Our working hypothesis is the formation of aparticulate domains and lamellar-to-hexagon{sub II} phase transitions in the plasma membrane and subtending lamellae are manifestations of hydration-dependent bilayer-bilayer interactions.

The cell wall protein Ecm33 of Candida albicans is involved in chronological life span, morphogenesis, cell wall regeneration, stress tolerance and host-cell interaction

Directory of Open Access Journals (Sweden)

Ana eGil-Bona

2016-02-01

Full Text Available Ecm33 is a glycosylphosphatidylinositol (GPI-anchored protein in the human pathogen Candida albicans. This protein is known to be involved in fungal cell wall integrity and is also critical for normal virulence in the mouse model of hematogenously disseminated candidiasis, but its function remains unknown. In this work, several phenotypic analyses of the C. albicans ecm33/ecm33 mutant (RML2U were performed. We observed that RML2U displays the inability of protoplast to regenerate the cell wall, activation of the cell wall integrity pathway, hypersensitivity to temperature, osmotic and oxidative stresses and a shortened chronological lifespan. During the exponential and stationary culture phases, nuclear and actin staining revealed the possible arrest of the cell cycle in RML2U cells. Interestingly, a veil growth, never previously described in C. albicans, was serendipitously observed under static stationary cells. The cells that formed this structure were also observed in cornmeal liquid cultures. These cells are giant, round cells, without DNA, and contain large vacuoles, similar to autophagic cells observed in other fungi. Furthermore, RML2U was phagocytozed more than the wild-type strain by macrophages at earlier time points, but the damage caused to the mouse cells was less than with the wild-type strain. Additionally, the percentage of RML2U apoptotic cells after interaction with macrophages was fewer than in the wild-type strain.