The present invention relates to a novel polynucleotide comprising SEQ ID NO. 1 useful for water-stress tolerance in biological systems, where the polynucleotide is differentially expressed in Tea plant under drought conditions.

1. Field of the Invention
The present invention relates to three novel genes of SEQ ID Nos. 1-3 useful for water-stress tolerance in biological systems, wherein said genes are differentially expressed in Tea plant under drought conditions and a method of introducing said genes into a biological system to help develop water stress tolerance.
2. Prior Art
Crop performance is sensitive to a number of biotic and abiotic factors, wherein drought stress constitutes an important yield-limiting determinant. Drought stress in context to the present invention refers to the situation when the amount of water in the plant is not sufficient to meet the transpirational requirements of the plant that leads to altered visible symptoms such as leaf curling. Drought should also be quantifiable through an important physiological parameter, leaf water potential (a measure of water status within the leaf tissue). Plant response to water deficit is dependent on the amount of water lost, the rate of loss, the duration of drought stress, the plant variety/species under consideration, developmental stage of the plant, and other environmental variables such as temperature, relative humidity etc.
Stress affects many metabolic pathways and structures, which may be the result of some up or down-regulated genes. Many of the water deficit induced genes encode gene products predicted to protect cellular function. One often noticed response of the plant is the accumulation of metabolically compatible solutes such as proline, glycine betaine, pinitol, carnitine, mannitol, sorbitol, polyols, trehalose, sucrose, oligosachharides and fructans in large quantities. These are chemically dissimilar and are excluded from the surface of the proteins, thus keeping the proteins preferentially hydrated. Accumulation of these compounds results in decreased water potential thus, facilitating water movement in the cell and helps in maintaining the turgor, a mechanism proposed to safeguard against water deficit.
These compounds have capability to (a) stabilize the membranes and other macromolecules such as nucleic acids and proteins, and can function as scavenger of free radicals. Indeed the transgenic plants over-expressing the genes responsible for the synthesis of these compounds were found to be more tolerant as compared to the wild types under the situation of water deficiency. Classical studies include: (a) transgenic tobacco overexpressing SacB gene (encoding levan-sucrase) from Bacillus subtilis accumulated fructan several folds that showed significantly greater growth and dry weight accumulation in response to drought stress (Pilon-Smits, E. A. H., Ebskarnp, M. J. M., Paul, M. J., Jeuken, M. J. W., Weisbeek, P. J. and Smeekens, S. C. M. (1995) Improved performance of transgenic fructan-accumulating tobacco under drought stress. Plant Physiol. 107:125-130); (b) Transgenic tobacco overexpressing P5CS (Δ1-pyrroline-5-carboxylate synthetase; the enzyme involved in the proline biosynthesis from L-glutamate via Δ1-pyrroline-5-carboxylate) from mothbean (Vigna aconitifolia) lead to 10-18 fold increase in proline content and showed better growth under water stress compared to the wild type (Kavi, K. P. B., Hong, Z., Miao, G-H., Hu C. A. A. and Verma, D. P. S.(1995) Plant Physiol. 108:1387-1394); (c) Transgeric tobacco expressing TPS1 gene (synthesizing trehalose-6-phosphate synthase) from yeast accumulated trehalose and showed better drought tolerance compared to the wild types (Holmstorm, K. O., Mantyla, E., Mandal, W. A., Palva, E. T., Tunnela, O. E. and Londesborough J. (1996) Drought tolerence in tobacco. Nature. 379: 683-684); (d) Transgenic tobacco overexpressing betB gene (synthesizing betaine aldehyde dehydrogenase) from E. coli showed better performance under osmotic stress conditions (Holmstrom, K. O., Welin, B. and Mandal, A. (1994) Production of the Escherichia coli betaine-aldehyde dehydrogenase an enzyme required for the synthesis of the osmoprotectant glycine betaine, in transgenic plants. Plant J. 6:749-758); (e) Transgenic tobacco expressing imtl gene (synthesizing myo-inositol-o-methyl transferase and involved in D-ononitol biosynthesis) from Mesembryanthemum crystallinum showed more adaptation to water stress (Sheveleva, E., Chmara, W., Bohnert, H. J. and Jensen, R. G. (1997) Increased salt and drought tolerence by D-Ononitol production in transgenic Nicotiana tabacum. Plant Physiol.5: 1211-1219); (f) Production of some of these osmo-protectants under drought stress is mediated through the plant hormone abscisic acid (ABA).
Recently, 9-cis-epoxycarotenoid dioxygenase gene (NCED), involved in ABA synthesis has been found to be strongly induced under water deficit in the 8-day-old cowpea plants (Iuchi, S., Kobayashi., Yamaguchi-Shinozaki, K. and Shinozaki, K. (2000) A stress-inducible gene for 9-cis-epoxycarotenoid dioygenase involved in abscisic acid biosynthesis under water stress in drought-tolerant cowpea. Plant physiol. 123:553-562). NCED mRNA was found to be increased both In reply to: water stressed leaves and roots of tomato (Thompson, A. J., Jackson, A. C., Parker, R. A., Morpeth, D. R., Burbidge, A. and Taylor, I. B. (2000) Abscisic acid biosynthesis in tomato: regulation of dioxygenase mRNA by light/dark cycles, water stress and abscisic acid. Plant. Mol. Biol. 42:833-845).
Apart from the osmolytes assisting in maintaining the hydration status, drought or osmotically stressed plants, synthesize several genes, which produce water channel proteins and water transport proteins such as membrane proteins of family aquaporins that can alter the cellular water potential and thus, protect against water deficit (Chrispeels, M. J. and Agre, P. (1994) Water channel proteins of plants and animal cells. Trends in Biochem Sci. 19:421-425; Bohnert H, J. and Jensen, R. G. (1996).
Strategies for engineering water-stress tolerance in plants. TIBTECH. 14:89-97; Johansson, I., Larsson, C., Ek B. and Kjellbom, P. (1996) The major integral proteins of spinach leaf plasma membranes are putative aquaporins and are phosphorylated in response to Ca and apoplastic water potential. The Plant Cell. 8:1181-1191. Accumulation of LEAs to high concentrations also coincides with the acquisition of desiccation tolerance. One of the groups-3 of LEA proteins is predicted to play a role in the sequestration of ions that are concentrated during cellular dehydration. Another group-5 of LEA proteins are predicted to sequester ions during water loss. The maintenance of total water potential during water deficit can be achieved by osmotic adjustment. Two proteins, osmotin and nonspecific lipid transfer proteins, are stress induced and are thought to play a role in controlling pathogens. Nonspecific lipid transfer proteins are induced by drought (Plant A. L., Cohen, A., Moses, M. S. and Bray, E. A. (1991) Nucleotide sequence and spatial expression pattern of a drought abscisic acid induced gene in tomato. Plant Physiol. 97:900-906; Toress-Schumann, S., Godoy, J. A. and Pintor-Toro, J. A. (1992) A probable lipid transfer protein is induced by NaCl in stems of tomato plants. Plant Mol. Biol.18:749-757).
Heat shock proteins that are induced by water deficit (Borkird, C., Simoens, C., Villarroel, R. and VanMontagu M. (1991) Gene associated with water-stress adaptation of rice cells and identification of two genes as hsp 70 and ubiquitin. Physiol. Plant. 82: 449-457; Almoguera, C. and Jordano, J. (1992) Developmental and environmental concurrent expression in sunflower dry-seed-stored low-molecular-weight heat shock protein and Lea mRNAs. Plant Mol. Biol. 19:781-792) may be involved in refolding of proteins in order to regain their function, or the prevention of protein aggregation (Vierling E. (1991) the roles of heat shock proteins in plants. Annual review of Plant Physiol. and Plant Mol. Biol. 42:579-620) during drought.
Small HSPs are another type of proteins those have been associated with plant desiccation tolerance. Small HSPs might act as molecular chaperones during seed dehydration and first few days of rehydration (Hoekstra, F, A., Golovina, E, A. and Buitink, J. (2001) Mechanisms of plant desiccation tolerance. Trends in Plant Science. 6(9): 43-439). OsHSP110 accumulated in shoots of rice seedlings in response to salinity, drought and low temperature apart from heat shock. It has been shown that two of the hsps, hsp70 in maize and hsp27 in soybean can also be induced by water stress (Sachs, M. M. and David Ho, T. H. (1986). Alterations of gene expression during environmental stress in plants. Ann. Rev. Plant Physiol. 37: 363-376). Most of the changes in gene expression occur during dehydration and thus many dehydration-specific gene products have been isolated but very few rehydration-specific proteins are known (Bernacchia, G., Schwall, G., Lottspeich, F., Salamini, F., and Bartels, D. (1995) Molecular Characterization of the Rehydration process in the Resurrection Plant Craterostigma Plantagineum. EMBO J 14: 610-618).
Complex regulatory and signaling processes, most of which are not understood, control the expression of genes during water deficit. Genes involved in two types of protein degrading mechanisms, proteases and ubiquitin are induced by water deficit. The gene products may be involved in degradation of proteins that are denatured during cellular water loss. Also, thiol protease an enzyme involved in degradation of proteins that have been denatured by stress, is induced by water deficit (Guerrero, F. D., Jones, J. T. and Mullet, J. E. (1990) Turgor-responsive gene transcription and RNA levels increases rapidly when pea shoots are wilted: sequence and expression of three inducible genes. Plant Mol. Biol. 15: 11-26).
Neale, A. D., Blomstedt, C. K., Bronson, P., Le, T.-N., Guthridge, K., Evans, J., Gaff, D. F. and Hamill, J. D. (2000. The isolation of genes from the resurrection grass Sporobulus stapfianus which are induced during severe drought stress. Plant, Cell and Environment. 23:265-277) isolated drought stress induced genes from resurrection grass Sporobolus stapfianus. Detected genes were found to encode an eIF1 translation initiation factor, two drought stress-inducible glycine-rich proteins, a tonoplast-intrinsic protein (TIP) and an early light-inducible protein (ELIP). Previously, no such gene products have been found to be associated with drought stress. This is the first report suggesting that a gene encoding an eLF1 translation initiation factor may have a role in the drought stress response of plants.
Several different stresses may trigger the same or similar signal transduction pathways. The plant hormone ABA also accumulates in response to the physical phenomenon of loss of water caused by the different stresses, and elevation in endogenous ABA content is known to induce certain water-deficit induced genes. Therefore, ABA accumulation is a step in one of the signal transduction pathways that induces genes during water deficit. Various protein kinases have been reported in plants and are thought to function in phosphorylation processes in various signal transduction pathways, including water-stress and ABA responses.
A cDNA, pKABA1, corresponding to a protein kinase, which is induced by ABA, has been isolated (Anderberg, R. J. and Walker-Simmons, M. K. (1992) Isolation of wheat cDNA clone for an abscisic acid-inducible trascript with homology to protein kinases.
Proc. Natl. Acad. Sci. USA 89: 10183-10187). A new homoebox-containing gene, Athb-12 and Athb-7 are induced by water deficit and exogenous ABA treatment but time course experiment have shown that both of these are regulated in a different manner (Lee, Y. H. and Chun. J. Y. (1998) A new homeodomain-leucine zipper gene from Arabidopsis thaliania induced by water stress and abscisic acid treatment. Plant Mol. Biol. 37: 377-384).
Available evidences suggest that stress induced responses may be ABA mediated or independent of ABA (Shinozaki, K. and Yamaguchi-Shinozaki, K. (1997) Gene expression and signal transduction in water-stress response. Plant Physiol. 115: 327-334). ABA mediated gene response may require or may not require protein synthesis to take place. The induction of mRNA of rd22 gene by ABA, which showed homology to an unidentified seed protein of Vicia faba, required protein synthesis to take place since cycloheximide inhibited induction of the gene (Yamaguchi-Shinozaki K. and Shinozaki, K. (1993) The plant hormone abscisic acid mediates the drought-induced expression but not the seed-specific expression of rd22, a gene responsive to dehydration stress in Arabidopsis thaliana. Mol. Gen. Genet. 238:17-25).
Structure analysis of the gene revealed the presence of regulatory sequences (cis-acting motif) asl (TGACG; SEQ ID NO: 4) and sp1 (GGGCGG; SEQ ID NO: 5) at −463 and −443 positions, respectively (Briggs, M. R., Kadonaga, J. T., Bell, S. P. and Tijan, R. (1986). Purification and Biochemical characterization of the promoter-specific transcription factor, Spl. Science 234:47-52; Lam, E., Benfey, P. N., Gilmartin, P. M., Fang, R. X. and Chua N-H. (1989). Site specific mutations alter in vitro factor binding and change promoter expression pattern in transgenic plants. Proc. Natl. Acad. Sci. USA. 86: 7890-7894). Also, were present the sequences that resembled myb (a family of transcription factors with Tip cluster motif) recognition elements TGGTTAG (SEQ ID NO: 6) at −144 and −666 and 2 bHLH (basic helix-loop-helix; MYC) recognition elements (CACATG; SEQ ID NO: 7) at −200 and −191 position. A cDNA (rd22BP1) encoding a MYC related DNA binding protein was isolated, which was found to encode a 68 kD protein that has a typical DNA binding domain of a basic region helix-loop-helix leucine zipper motif in MYC-related transcription factors.
The protein indeed binds to the MYC recognition site (Abe, H., Yamaguchi-Shinozaki, K., Urao, T., Iwasaki, T., Hosokawa, D. and Shinozaki, K. (1997) Role of Arabidopsis MYC and MYB Homologs in Drought- and Abscisic Acid-Regulated Gene Expression. The Plant Cell. 9:1859-1868). A drought and ABA inducible gene has also been cloned that encodes MYB-related protein ATMYB2. Both rd22BP 1 (MYC) and ATMYB2 (MYB) proteins were shown to function as transcription activators in the dehydration and ABA-inducible expression of the rd22 gene (Abe, H., Yamaguchi -Shinozaki, K., Urao, T., Iwasaki, T., Hosokawa, D and Shinozaki, K. (1997). Role of Arabidopsis WC and MYB Homologs in Drought- and Abscisic Acid-Regulated Gene Expression. The Plant Cell. 9:1859-1868).
In contrast to rd22 in Arabidopsis, HVA22 gene in barley is induced in response to drought and ABA, but is also induced in the presence of cycloheximide. The promoter region of HVA22 contains ABA responsive complex ABRE3, CE1 and another ABA responsive complex that relies on the interaction of a G-box with another yet unidentified coupling element (Shen, Q. and Ho, T-H D. (1995) Functional Dissection of an Abscisic Acid (ABA)-Inducible gene Reveals Two Independent ABA-Responsive Complexes Each Containing a G-Box and a novel cis-Acting Element. The Plant Cell. 7:295-307)
Yamaguchi-Shinozaki K and Shinozaki, K. (1993. Characterization of the expression of dessication-responsive rd29 gene of Arabidopsis thaliana and analysis of its promoter in transgenic plants. Mol. Gen. Genet. 236: 331-340) cloned a dehydration responsive gene rd29A that was independent of ABA responsive pathway. The sequence TACCGACAT (SEQ ID NO: 8) was found to be regulating the genes induced under drought conditions and was found in the promoter regions of other dehydration inducible genes.
Upon over-expression of DREBIA (a dehydration responsive element binding protein) under the control of rd29a promoter in A. thaliana, a number of stress tolerant genes were expressed and resulted in an improved tolerance under drought and several other stresses (Kasuga, M., Liu, Q., Miura, S., Yamaguchi-Shinozaki, K. and Shinozaki, K. (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nature Biotechnology. 17:287-291).
Analysis of another gene of DRE-binding protein DREB2 showed that its promoter was induced under water stress in transgenic arabidopsis (Nakasimha, K., Shinwari, Z. K., Sakuma, Y., Seki, M., Miura, S., Shinozaki, K. and Yamaguchi-Shinozaki, K.(2000) Organization and expression of two Alabidopsis DREB2 genes encoding DRE-binding proteins involved in dehydration and high salinity responsive gene expression. Plant. Mol. Biol. 42:657-665). These genes do not require ABA for their expression, but do respond to exogenous ABA.
There are also drought inducible genes that do not respond to ABA treatment. These include rd 21, erd1, and rd 19 that code for thiol proteases, CIp protease and thiol protease, respectively (Shinozaki, K. and Yamaguchi-Shinozaki, K. (1997) Gene expression and signal transduction in water-stress response. Plant Physiol. 115:327-334). Indeed, the information on such genes is very scarce.
There is always a need and search for novel drought related genes so that better adaptation may be sought. Apart from the genes and gene sequences listed in the Table 1, the novel gene sequences may be listed as follows:
ABRE. ABA-responsive element (PyACGTGGC; SEQ ID NO: 9) (Shen Q and Ho (1995) Functional Dissection of an Abscisic Acid (ABA)-Inducible gene Reveals Two Independent ABA-Responsive Complexes Each Containing a G-Box and a novel cis-Acting Element. The Plant Cell. 7:295-307).
G-box, ubiquitous regulatory elements (CACGTG; SEQ ID NO: 10). (Menkens, A. E., Schindler, U. and Cashmore A. R. (1995) The G-box: ubiquitous regulatory DNA element in plants bound by GBF family of bZIP proteins. Trends in Biochem Sci. 20:506-510).
DRE, Dehydration-responsive element (TACCGACAT; SEQ ID NO: 11) (Shinozaki, K. and Yamaguchi-Shinozaki, K. (1996) Molecular responses to drought and cold stress. Current opinion in biotechnology. 7:161-167). MYBRS, MYB recognition sequence (PyAACPyPu; SEQ ID NO: 12) (Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. The Plant Cell 5:1529-1539).
MYCRS, MYC recognition sequence (CANNTG; SEQ ID NO: 13) (Abe, H., Yamaguchi-Shinozali, K., Urao, T., Iwasaki, T., Hosokawa, D and Shinozaki, K. (1997) Role of Arabidopsis MYC and MYB Homologs in Drought- and Abscisic Acid-Regulated Gene Expression. The Plant Cell. 9: 1859-1868).
While working with gene(s) and gene fragments (gene fragment in context to the present invention refers to partial nucleotide sequences of the complete gene), related to drought or other stresses, the following are possibilities:
(a) Gene can be Cloned Through Several Routs as Shown Below in Table 1.
TABLE 1Route/TechniqueUsedReferenceProtein sequencingWeretilnyk, E. A. and Hanson, A. D. 1990. Molecular cloning offollowed by oligo-a plant betenin oldohydo dehydrogenase, an enzyme implicatednucleotide synthesisin soaptation to salinity and drought Prod. Notl. Acad. Sci. IISAand screening87: 2745-2749.Plaque hybridizationNakashima, k. Shinwari, Z. K., Sakuma, Y., Seki, M., Miura, S.,Shipozaki, K and Yamaguchi-Shinozaki, K. 2000. Organizationand expression of two Arapldopsts DRED2 genes eneedingDRE binding proteins involved in dehydration and high salinityresponsive gene expression Plant. Mol. Biol. 42. 657-665.PCR based cloningHirayama, T., Ohto, C., Mizoguchi, T., and Shinozaki, K. 1995.A gene encoding a phosphoinositol-specific phospholipase C ininduced by dehydration and salt stress in Arabidopsis thalianaProc. Natl Acad. Sci. 92: 3903-3907Library screening usingRichard, S., Morency, M., Drevet, C., Jouanin, L., and Seguin, S.heterologous probe2000. Isolation and characterization of a dehydrin gene fromwhite spruce induced upon wounding, drought and cold stress.Plant Mol. Biol. 43: 1-10.Gene cloning usingRoberts, J. K. and Key, J. L. 1991. Isolation and characterizationheterologous probeof a soybean hsp 70 gene. Plant molecular biology, 16: 671-683.Differential ScreeningChang, S., Puryear, J. D., Dias, A. A. D. L, Funkhouser, E. A.,Newton, R. J., and Calway, J. 1996. Gene expression underwater dificit in lobiolly pine (Pinus taeda): isolation andcharacterization of cDNA clones. Physiol. Plant. 97: 139-148.MicroarraySeki, M., Nerusaka, M., Abe, H., Kasuga, M., Yamaguchi-Shinozaki, K., Caminci, P. Hayashizaki, Y., and Shinozaki, K.2001 Plant Cell 113: 61-72Subtractivea. Lee, S. W., Tomasetto, C., and Sagar R, 1991. Positivehybridizationselection of fumous suppression genes by subtractivehybridization Proc. Nati Acad. Sci. USA, 88: 2825-2829.b. Buchanan-Wollaston, V. and Ainaworth, C, 1997 Leafsenescence in Brassica naus cloning of senescence relatedgene bu substractive hybridication Plant Mol. Biol. 33, 821-834.(b) Gene Cloned from Organisms can be Expressed in Other Organisms.
As has been shown by Kishor, Kavi. P. B. R. Hong, Z., Miao, G. H., Hu, C. A. and Verma, D. P. S. (1995 Overexpression fo pyrroline-5-carboxylate synthetase increases proline production and confers osmotalerence in transgenic plants. Plant Physiol. 108:1387-1394 and the references therein) that the gene pyrroline-5-carboxylase synthetase was cloned from Vigna aconotifolia and expressed into tobacco through transgenic technology Transgenic tobacco plants were more tolerant under water stress conditions.
Pilon Smits, E. A. H., Ebskamp. M. J. M., Paul, M. J. Jeuken, M. J. W., Weisbeek, P. J. and Smeekens. Improved performance of transgenic fructan-accumulating tobacco under drought stress. Plant. Physiol. 107:125-130) transferred SacB gene from Bacillus subtilis into tobacco and found increased drought tolerance.
Holmstrom, K. O., Welin, B. and Mandal, A. (1994, Production of the Escherichia coli betaine-aidehyde sakydrogonase an enzyme required for the synthesis of the osmoprotectant glycine betaine, in transgenic plants. Plant J. 6:749-758) transferred betaine-aldehyde dehydrogenase from Escherichia coli (a microorganism) into tobacco (higher plant) and found to be drought tolerant.
(c) Genes Expressed in Response to Drought Stress can be Expressed by Other Environmental Variables as Well.
Iuchi, S., Kobayashi, Yamaguchi-Shimuzaki, K and Shinozaki, Kazuo (2000 A stress-inducibe gene for 9-cis-epoxycarotenoid dioxygenase involved in abscisic acid biosynthesis under water stress in drought tolerant compound. Plant physical 123:553-662) reported the expression of VcNCED1 in response to water and salt stress.
Pelloux J., Jolivet, Y., Hontaine, V., Banvoy, J., and Dizengromel, P. (2001 Changen in Rubieco and Rubisco activase gene expression and polypeptide expression.
Richard. S. Mordancy. M. Drevet. C. Jouanin, L. and Seguin, S. (2000. Isolation and characterization of a dehydrin gene from white spruce induced upon wounding, drought and cold stress. Plant Mol. Biol. 43: -1-10} reported a gene PgDhnl. which was induced In repose to drought, cold stress and upon wounding.
Nakashima. K. Shinwari, Z. K. Sakuma, Y. Seki. M. Miura, S, Shinozaki, K. and Yamaguchi-binozaki K. (2000 Organization and expression of two Arabidopsis DREB2 genes encoding DRE-binding proteins involved in dehydration and high salinity responsive gene expression. Plant. Mol, Biol 42; 657-665) reported the expression of drought responsive element DREB2 genes in response to dehydration and high salinity stress.
Hirayama. T. Ohto. C. Mizoguchi. T. and Shinozaki, K. (1995. A gene encoding a phosphoinositol-specific phospholipase C in induced by dehydration and salt stress in Arabidopsis thaliano, Proc. Natl. Acad. Sci 92: 3903-3907) reported expression of phosphoinosltol-specific phospholipase C in response to drought, salinity and low temperature.
Weretilnyk. E. A., and Hanson. A. D. (1990. Molecular cloning of a plant betaine-aldehyde dehydrogenase, an enzyme implicated in adaptation to salinity and drought. Proc. Natl. Acad. Sci., USA, 87: 2745-2749) reported the expression of betaine-aldehyde dehydrogenase gene in response to drought as well salinity,
D. Identified Gene may be Used to Study Regulatory Elements. Regulatory Elements in Context to Present Invention Relate to the Regions such as Promoters, Transcriptional Factors and Other Sequences which Control the Expression of the Gene.
Using stress regulated gene HVA1. Straub, P. P. Shen Q. and Ho, Tuan-hua. D. (1994. Structure and promoter analysis of an ABA- and stress-regulated barley gene, HVA1. Plant. Mol. Biol. 26: 617-630) analysed promoter of the gene, Michel, D., Salamini F., Bartels, D. Dale, P., Baga, M. and szalay, A. (1993. Analysis of a desiccation and ABA-responsive promoter Isolated from the resurrection plant Craterostigma plantagineum Plant Journal 4: 29-40) selected drought responsive gene CdeT27-46 and analysed its promoter region.
Urao T, Yamaguchi-Shinozaki K. Urao S. Shinozaki K. (1993. An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell 5: 1529-1539) identified the sequences encoding transcription factors in a dehydration responsive gene Atmyb2 Yamaguchi-Shinozaki. K. and Shinozaki, K. (1997, Characterization of the expression of a desiccation-responsive rd29 gene of arabidopsis thaliana and analysis of its promoter in transgenic plants. Mol Gen Genet 236: 331-340) analysed the promoter region of a drought inducible gene rc(29.
Abe. H., Yamaguchi-Shinozaki. K. Urao, T. Iwasaki, T. Hosokawa. D and Shinozaki. K. (1997. Role of Arabidopsis MYC and MYB Homologs in Drought- and Abscisic Acid-Regulated Gene Expression. The Plant Cell.9: 1859-1868) analysed a drought inducible gene rd22 for regulatory factors.
Shen Q and Ho T. D. (1995. Functional Dissection of an Abscisic Acid (ABA)-Inducible gene Reveals Two Independent ABA-Responsive Complexes Each Containing a G-Box and a novel c/s-Acting Element. The Plant Cell. 7: 295-307) analysed HVA22 gene for regulatory elements and reported novel coupling elements.
e. Gene or Gene Fragment Isolated from One System can be Used as a Probe to Study the Similar Genes in Other Plant Systems
Roberts, J K and Key. J. L. (1991. Isolation and characterization of a soybean hsp70 gene. Plant molecular biology, 16: 671-683) used hsp70 gene cloned from Drosophila to clone the similar gene from soybean.
Singia. S. L. Pareek A and Grover. (1997 Yeast HSP104 homologue rice HSP 110 is developmental- and stress regulated Plant science, 125: 211-219) showed that yeast hsp KM and rice hsp 110 are very similar. These are expressed in response to desiccation, salinity, low temperature and high temperature.
Shen, Q. Chen, C. N. Brands. A. Pan. S. M. and Ho, T. D. (2001 The stress- and abscisic acid-induced barley gene HVA 22: developmental regulation and homologues in diverse organisms. Plant Molecular Biology. 45: 327-340) reported a drought inducible gene HVA 22 in several organisms such as cereals, arabidopsis. Caenorhabitis elegans. man. mouse, and yeast.
(f) Gene Expressed in One Organ can be Expressed in Organ as Well as Shown Below in Table 2:
TABLE 2OrganReferenceRoots andNemoto, Y., Kawakami, N., and Sasakuma. 1999. Isolationleavesof novel early salt-responding genes from wheat (Triticumaestivum L.) by differential display. Theor. Appl. Genet 98:673-678Thompson, A. J., Jackson, A. C., Parker, R. A., Morpeth, D. R.,Burbidge, A. and Taylor, I. B. (2000) Abscisic acidbiosynthesis in tomato: regulation of dioxygenase mRNAby light/dark cycles, water stress and abscisic acid.Plant.Mol.Biol. 42: 833-845Sheaths andClaes, B., Dekkkeyser, R., Villarroel, R., Bulcke, M. V. D., Bauw, G.,rootsMontagu, M. V., and Caplan, A. 1990. Characterization of a ricegene showing organ specific expression in response to salt stressand drought. Plant Cell, 2: 19-27.Stem TissueRichard, S., Morency, M., Drevet, C., Jouanin, L., and Seguin, S.and partially2000. Isolation and characterization of a dehydrin gene from whiteexpandedspruce induced upon wounding, drought and cold stress, Plant Mol.vegetative buds,Biol.43: 1-10reproductivebuds(g) It is possible to Clone Full Length cDNAs or Genomic DNA by Using Standard Protocols as detailed by Ausubel. F. M. Brend R. Kingston. R. E., Moore. D. D. Seidman. J. G. Smith, J. A., Struhl. K. 1987. Current protocols in molecular biology. Publisher John Wiley and Sons. New York: and Sambrook. J. Fritsch, E. F, and Maniatis. T. 1989. Molecular cloning; a laboratory manual, Second edition. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
A summary of various drought-related genes is given below in Table 3.
TABLE 3List of drought related genes along with their source and the predicted function.PredictedSpecies from whichGENESrole/HomologyisolatedREFRENCEA1494Cysteine thiolArabidopsis thalianaWilliams et al.,protease1994 Plant Mol.Biol. 25: 259-270ADHAlcohol″de Bruxelles et al.,dehydrogenase1996 Plant Physiol.111: 381-391;Dolferus et al.,1994 Plant Physiol.105: 1075-1087;Jarillo et al., 1993Plant Physiol. 833-837Athb-7Homeodomain″Söderman et al.,leucine zipper1996 Plant Journaltransfactor10: 375-381Athb-12Homeodomain″Lee and Chun, 1998leucine zipperPlant Mol. Biol. 37:transfactor377-384AthH2Aquaporin″Kaldenhoff et al.,1993 Plant Mol.Biol. 23: 1187-1198AthK1Histidine kinase″Urao et al., 1998FEBS Left. 427:175-178CDPK1/K2Cal dependent″Urao et al., 1994protein kinaseMol. Gen. Genet.244: 331-340HSP70-1/ERD2HSP-cognate″Kiyosue et al., 1994Plant Mol. Biol. 25:791-798HSP81-2/ERD8HSP-cognate″Kiyosue et al., 1994Plant Mol. Biol. 25:791-798rd22Unidentified seed″Yamaguchi-protein of ViciaShinozaki andfabaShinozaki., 1993Mol. Gen. Genet.238: 17-25RAB18Dehydrin″Lang et al., 1994Plant Physiol. 104:1341-1349; Langand Palva, 1992Plant Mol. Biol. 20:951-962RD19Cysteine protease″Koizumi et al.,1993 Gene 129:175-182RD28, RD21Cysteine protease″Yamaguchi-Shinozaki et al.,1992 Plant CellPhysiol. 33: 217-224rd29A, rd29BDrought responsive″Iwasaki et al., 1997promoter element,Plant Physiol. 115:drought related1287; Wang et al.,genes1995 Plant Mol.Biol. 28: 605-617DREB1ADehydration″Kasuga et al, 1999responsiveNatureelements bindingBiotechnology. 17:proteins287-291Tps1Trehalose″Holmstrom et al.,biosynthesis1994 Plant Journal6: 749-758RPK1Receptor-like″Hong et al., 1997protein kinasePlant Physiol 113:1203-1212cAtP5CSΔ1-pyrroline-5-″Yoshiba et al., 1995carboxylatePlant J. 7: 751-760synthetaserd19A; rd21ACysteine proteases″Koizumi et al.,1993 Gene129: 175-182UBQIUbiquitin extension″Kiyosue et al., 1994proteinPlant Mol. Biol. 25:791-798cATCDPK1; cATCDPK2CA2+-dependent,″Urao et al., 1994calmodulin-The Plant Cell 5:independent1429-439protein kinasescAtPLC1Phosphatidylinositol-″Hirayama et al.,specific1995 Proc. Natl.phospholipase CAcad. Sci. USA 92:3903-3907ERD11; ERD13Glutathione S-″Kiyosue et al., 1993.transferasesBiochem. Biophys.Res. Comm.196: 1214-1220cAtsEHSoluble epoxide″Kiyosue et al., 1994hydrolasePlant J. 6: 259-269kin2Similarity to″Kurkela & Borg-animal antifreezeFranck 1992 PlantproteinsMol. Biol. 29: 689-692pA1494Similarity to″Williams et al.,proteases1994 Plant Mol.Biol. 25: 259-270ERD1Similar to a Clp″Kiyosue et al., 1993ATP-dependentBiochem. Biophys.protease subunitRes. Comm.196: 1214-1220Athsp70-1Similar to the″Kiyosue et al.,HSP70 heat-shock-1994 Plant Mol.protein familyBiol. 25: 791-98Athsp81-2similar to the″Kiyosue et al., 1994HSP81 heat-shockPlant Mol. Biol.protein family25: 791-98rd22Similar to an″Iwasaki et al., 1995unidentified seedMol. Gen. Genet.protein from Vicia247: 391-398fabalti65, lti78Unknown″Nordin et al., 1993Plant Mol. Biol. 21:641-653pRABAT1D11 LEA-protein″Lang & Palva,related1992 Plant Mol.Biol. 20: 951-962Atmyb2MYB-protein-″Urao et al., 1993relatedThe Plant Cell 5:transcription factor1429-1439ERD10; ERD14D11 LEA-protein″Kiyosue et al., 1994relatedThe Plant CellPhysiol. 35: 225-231SacBFructosylBacillus subtilisPilon-Smits et al.,transferase1995 Plant Physiol.107: 125-130MC12LKR/SDH cDNABrassica napusDeleu et al., 1999of A. thlianaPlant Cell andEnvironment 22:979-988MC43His-3 linker″Deleu et al., 1999protein/ribosomalPlant Cell andprotein S12Environment 22:979-988pBN115Similar toBrassica napusWeretilnyk et al.,polypeptides1993 Plant Physiol.encoded by pBN19101: 171-177and pNB26 (B. napus),and COR15(A. thaliana)BnD22Similar to proteaseBrassica napusDowning et al.,inhibitors1992 Plant J. 2:685-693VuNCED1ABA biosynthesisCowpeaIuchi et al., 2000Plant Physiol. 123:553-562GapC-CratCytosolicCraterostigma plantagineumVelasco et al., 1994glyceraldehyde 3-Plant Mol. Biol. 26:phosphate541-546dehydrogenasepSPS1Sucrose-phosphate″Ingrams & Bartels,synthase1996 Annu RevPlant Physiol 47:377-403PSS1; pSS2Sucrose synthases″Ingrams & Bartels,1996 Annu RevPlant Physiol 47:377-403pcC 37-31Similar to early-″Bartels et al., 1992light-inducibleEMBO J. 11: 2771-2778proteinspcC 13-62Unknown″Piatkowski et al.,1990 Plant Physiol.94: 1682-1688pcC 27-04D11 LEA-protein″Piatkowski et al.,related1990 Plant Physiol.94: 1682-1688pcC 6-19D11 LEA-protein″Piatkowski et al.,related1990 Plant Physiol.94: 1682-1688pcC 3-06D7 LEA-protein″Piatkowski et al.,related1990 Plant Physiol.94: 1682-1688pcC 17-45D95 LEA-protein″Piatkowski et al.,related1990 Plant Physiol.94: 1682-1688pcECP40D11 LEA-proteinDaucus carotaKiyosue et al., 1993relatedPlant Mol. Biol.21: 1053-1068Bet BGlycine betaineEscherichia coliHolmstrom el al.,biosynthesis1994 Plant Journal6: 749-758pTS.6Plasma membraneGlycine maxSurowy and Boyer,H+-ATPase1991 Plant. Mol.Biol 16: 251-262SC514Lipoxygenase″Bell and Mullet1991 Mol. Gen.Genet. 230: 456-462Ha hsp17.6HaLow-molecular-Helianthus annuusCoca et al., 1994hsp17.9weight heat-shockPlant Mol. Biol. 25:proteins479-492Ha ds 10D19 LEA-protein″Almoguera andrelatedJordano1992 PlantMol. Biol. 19: 781-792Ha ds11D113 LEA-protein″Almoguera andrelatedJordano, 1992 PlantMol. Biol. 19: 781-792B8; B9; B17D11 LEA-proteinHordeum vulgareClose et al., 1989relatedPlant Mol. Biol. 13:95-108B19.1; B19.3; B19.4D19 LEA-protein″Espelund et al., 1992relatedThe Plant CellEnviron. 18: 943-949HVA22LEA″Shen et al., 2001(Lateembryogenesis-Plant Mol. Biol. 45:abundant) and RAB327-340(responsive to ABA)BLT4Similar to protease″Dunn et al., 1991inhibitorsMol. Gen. Genet.229-389-394pBADBetaine aldehydeHordeum vulgareIshitani et al., 1995dehydrogenaseMol. Gen. Genet.247: 391-398pcht28Acidic endochitinaseLycopersicon chilenseChen et al., 1994Mol. Gen. Genet.145: 195-202SAM1; SAM3S-adenosyl-L-Lycopersicon esculentumEspartero et al., 1994methioninePlant Mol. Biol. 25:synthetases217-227P31Cytosolic copper/zinc″Perl-Treves andsuperoxide dismutaseGalun 1991 PlantMol. Biol. 17: 745-760TSW12A lipid transfer″Torres-Schumann etproteinal., 1992 Plant Mol.Biol. 18: 749-757pLE16Similar to lipid″Plant et al., 1991transfer proteinsPlant Physiol. 97:900-906pLE4D11 LEA-protein″Cohen et al., 1991relatedPlant Physiol. 97:1367-1374pUM90-1Similar to MsaciAMedicago sativaLuo et al., 1992 J.and pSM2075Biol. Chem. 267(22):polypeptides15367-15374pSM1075Similar to MsaciA″Luo et al., 1991 Plantand pUM90-1Mol. Biol. 17: 1267-1269polypeptidesMsaciASimilar to pUM90-1″Laberge et al., 1993and pSM2075Plant Physiol.polypeptides101: 1411-1412pPPC1PhosphoenolpyruvateMesembryanthemumVernon et al., 1993carboxylasecrystallinumThe Plant CellEnviron. 16: 437-444pRAB 16AD11 LEA-proteinOryza sativaMundy & Chua 1988.relatedEMBO J. 7: 2279-2286salTUnknown″Claes et al., 1990 ThePlant Cell 2: 19-27Apx1 geneCytosolic ascorbatePisum sativumMittler and Zilinskasperoxidase1994 Plant J. 5: 397-405Sod 2 geneCytosolic copper/zinc″White and Zilinskassuperoxide dismutase1991 Plant Physiol.96: 1291-129226gSome similarity to″Guerrero et al., 1990aldehydePlant Mol. Biol.dehydrogenase15: 11-267aSimilar to channel″Guerrero et al., 1990proteinsPlant Mol. Biol. 15:11-2615aSimilarity to″Guerrero et al., 1990proteasesPlant Mol. Biol. 15:11-26pLP2S-AdenosylPinus taedaChang et al., 1996methioninePhysiol. Plant. 97:synthatase139-148pLP3Silk fibrion and rat″Chang et al., 1996chondroitin corePhysiol. Plant. 97:protein139-148pLP4Tomato protein TMA″Chang et al., 1996SN1(water deficitPhysiol. Plant. 97:inducible)139-148pLP5Copper binding″Chang et al., 1996proteinPhysiol. Plant. 97:139-148P22Similar to proteaseRaphanus sativusLopez et al., 1994inhibitorsPhysiol. Plant. 91:605-614H26D11 LEA-proteinStellaria longipesRobertson andrelatedChandler 1992 PlantMol. Biol. 19: 1031-1044pMA2005D71 LEA-proteinTriticum aestivumCurry et al., 1991relatedPlant Mol. Biol.16: 1073-1076pMA1949D7 LEA-protein″Curry & Walker-relatedSimmons 1993 PlantMol. Biol. 21: 907-912EmD19 LEA-protein″Litts et al., 1987relatedNucleic Acids Res.15: 3607-3618PKABAIProtein kinase″Aderberg andWalker-Simmons1992 Proc. Natl.Acad. Sci. USA89: 10183-10187Pmbm1L-isoaspartyl″Mudgett & Clarkemethyltransferase1994J.Biol.Chem.269:25605-25612M3 (RAB-17)D11 LEA-proteinZea maysClose et al., 1989relatedPlant Mol. Biol. 13:95-108pMAH9Similar to RNA-″Gómez et al., 1988binding proteinsNature 334: 262-264
Below is specifically given a state of art knowledge with reference to cloning of drought stress related genes.
Reference may be made to document (1) by Yamaguchi-Shinozaki, K. and Shinozaki, K. (1994) The Plant Cell. 6: 251-264, wherein is described the identification of a novel cis-acting element involved in responsiveness to drought, low temperature; or high salt stress from a model plant Arabidopsis.
Reference may be made to document (2) by Li, L. g., Li, S. f., Tao, Y., and Kitagawa, Y. (2000) Plant Science 154: 43-51, wherein a novel water channel protein was cloned from rice which was shown to be involved with the chilling tolerance in Xenopus oocytes.
Reference may be made to document (3) by Tabaeizadeh; Zohrer,; Yu; Long-Xi; Chen; Ri-Dong, U.S. Pat. No. 5,656,474 dated Aug. 12, (1997) wherein two osmotic stress- and ABA-responsive members of the endochitinase gene family were isolated and identified from the leaves of drought-stressed Lycopersicon chilense plants.
Reference may be made to document (4) by Kim; Soo Young U.S. Pat. No. 6,245,905 dated Jun. 21, (2001) wherein a nucleic acid molecule encoding the Abscisic acid responsive element binding factor 2 (ABF2) was isolated that binds abscisic acid responsive elements in plants.
Reference may be made to document (5) by Kim; Soo Young U.S. Pat. No. 6,218,527 dated Apr. 17, (2001) wherein a nucleic acid molecule encoding the Abscisic acid responsive element binding factor 3 (ABF3) was isolated that binds abscisic acid responsive elements in plants.
Reference may be made to document (6) by Thomshow; Michael F.; Stockinger; Eric, J. U.S. Pat. No. 5,892,009 dated Apr. 6, 1999 wherein a gene designated as CBF1, encoding a protein CBF1, which binds to a region regulating expression of gene which promote cold temperature and dehydration tolerance in plants was cloned.
Reference may be made to document (7) by Chun; Jong-Yoon; Lee; Yong-Hun. U.S. Pat. No. 5,981,729 dated Nov. 9, 1999 wherein a novel gene induced by water deficit and abscisic acid was cloned.
The Drawbacks in the Prior Art are:
    a. Earlier work to clone the genes related to drought stress focused on model plant system and mainly annuals. Perennial evergreen plants such tea experience several rounds of drought stress during their life cycle. The plant is, therefore, expected to harbor novel gene(s) imparting tolerance to drought.    b. There is always search of novel genes so as to exploit it for generating more drought tolerant plants. Model plants such as Arabidopsis thaliana and other domesticated plants as mentioned in Table 1 have been used to clone the drought-related genes. Novel genes can be expected from a hitherto unstudied plant.    c. Methods reported to clone drought related gene relied on differential screening of cDNA library, analysis of differential cDNA library, and subtractive hybridization (Tables 1, 2 and 3). These have inherent limitation of using two samples at a time for analysis Therefore, after identification and cloning of differentially expressed genes, these used to be tested for their expression analysis during recovery and/or in response to other variables such as salt stress/ABA treatment etc. Therefore, appropriate technology needs to applied in order to focus on the desired gene at the beginning itself.
The above drawbacks have been eliminated for the first time in a simple and reliable manner by the present invention, which is not so obvious to the person skilled in the art.

Polynucleotide from a novel gene from drought stress tolerant tea plant

Council of Scientific and Industrial Research


By reducing thickness of a slidable contact member which is attached to an output shaft and a sector gear part and slidably contacts with a gear case, a wiper motor is downsized. A motive power conversion member comprises: an arm part rotatably connected to the worm wheel; and a sector gear part meshing with the output gear, wherein a gear shaft of the sector gear part is swingably supported with respect to the output shaft by a swinging plate. A first slidable contact member is mounted on a tip of the output shaft, and a second slidable contact part which is formed separately from the first slidable contact member is attached to a tip of the gear shaft, wherein the first slidable contact member and the second slidable contact part slidably contact with a cover of the gear case.

The present invention relates to a wiper motor, which has an electric motor and an output shaft for outputting motive power generated by the electric motor and swings a wiper arm attached to the output shaft.
In a rear wiper device or the like for wiping a rear window glass of a vehicle, a wiper motor having a motive power conversion mechanism for converting rotary motion of the electric motor into swinging motion of the output shaft is used to directly fix the wiper arm to the output shaft.
Such a wiper motor has a gear case comprising a case body formed into a bathtub-like shape and a cover for closing the case body, wherein a worm wheel of a worm gear mechanism for reducing the rotation of the electric motor and an output gear fixed to the output shaft are rotatably housed in the case body. A linking shaft is provided at the worm wheel away from a rotation-central axis, and an arm part serving as a motive power conversion member is rotatably connected to the linking shaft. In addition, a sector gear part meshing with the output gear is provided in the motive power conversion member, and a gear shaft which is press-fitted in an axial center of the sector gear part is rotatably supported by a swinging plate swingably supported by the output shaft. For this reason, when the electric motor operates and the worm wheel rotates, the arm part reciprocates and the sector gear part swings around the output gear, whereby the rotary motion of the worm wheel is converted into the swinging motion of the output shaft by the motive power conversion member.
In such a wiper motor, when an external force due to, for example, mischief is applied to the wiper arm or while the electric motor is operated, a load in a direction of separating these gears is generated between the output gear and the sector gear part. Then, due to the load, the gear shaft is inclined with respect to the output shaft which is supported by the gear case so that the swinging plate serves as a supporting point, whereby there is such fear that mesh of the respective gears may be misaligned. Therefore, for example in the wiper motor disclosed in Japanese Patent Laid-Open Publication No. 2000-341906, a resin-made slidable contact member is mounted on tip portions of the output shaft and the gear shaft to mutually connect the respective shafts and the slidable contact member slidably contacts with the cover, so that axial-directional movement of the respective shafts is restricted and the mesh of the respective gears is maintained.


The subject invention discloses a pneumatic tire having an outer circumferential tread wherein said tread is a sulfur-cured rubber composition comprised of (a) an isoprene-butadiene diblock rubber, said isoprene-butadiene diblock rubber being comprised of a butadiene block and an isoprene-butadiene block, wherein said butadiene block has a number average molecular weight which is within the range of about 25,000 to about 350,000, wherein said isoprene-butadiene block has a number average molecular weight which is within the range of about 25,000 to about 350,000, wherein said isoprene-butadiene diblock rubber has essentially one glass transition temperature which is within the range of about -100.degree. C. to about -70.degree. C., wherein said isoprene-butadiene diblock polymer has a Mooney ML-4 viscosity at 100.degree. C. which is within the range of about 50 to about 140, and wherein the repeat units derived from isoprene and 1,3-butadiene in the isoprene-butadiene block are in essentially random order; and (b) a second rubber selected from the group consisting of high vinyl polybutadiene rubber, styrene-isoprene-butadiene rubber, solution styrene-butadiene rubber and emulsion styrene-butadiene rubber.

The replacement cost of tires is one of the major expenses encountered by the trucking industry. Tire replacement cost and frequency is, of course, also of concern to most automobile and light truck owners. In recent years, many modifications have been implemented to improve the treadwear characteristics of tires. However, improvements in tire treadwear characteristics have sometimes been achieved by compromising the traction and/or rolling resistance characteristics of the tire.
In order to reduce the rolling resistance of a tire, rubbers having a high rebound can be utilized in making the tires' treads. Tires made with such rubbers undergo less energy loss during rolling and normally also exhibit improved treadwear characteristics. The traditional problem associated with this approach is that the tire's wet traction and wet skid resistance characteristics are compromised. This is because good rolling resistance which favors low energy loss and good traction characteristics which favor high energy loss are viscoelastically inconsistent properties.
In order to balance these two viscoelastically inconsistent properties, mixtures of various types of synthetic and natural rubber are normally utilized in tire treads. For instance, various mixtures of styrene-butadiene rubber and polybutadiene rubber are commonly used as a rubbery material for automobile tire treads. However, such blends are not totally satisfactory for all purposes.
U.S. Pat. No. 4,843,120 discloses that tires having improved performance characteristics can be prepared by utilizing rubbery polymers having multiple glass transition temperatures as the tread rubber. These rubbery polymers having multiple glass transition temperatures exhibit a first glass transition temperature which is within the range of about -110.degree. C. to -20.degree. C. and exhibit a second glass transition temperature which is within the range of about -50.degree. C. to 0.degree. C. According to U.S. Pat. No. 4,843,120, these polymers are made by polymerizing at least one conjugated diolefin monomer in a first reaction zone at a temperature and under conditions sufficient to produce a first polymeric segment having a glass transition temperature which is between -110.degree. C. and -20.degree. C. and subsequently continuing said polymerization in a second reaction zone at a temperature and under conditions sufficient to produce a second polymeric segment having a glass transition temperature which is between -20.degree. C. and 20.degree. C. Such polymerizations are normally catalyzed with an organolithium catalyst and are normally carried out in an inert organic solvent.
U.S. Pat. No. 5,137,998 discloses a process for preparing a rubbery terpolymer of styrene, isoprene and butadiene having multiple glass transition temperatures and having an excellent combination of properties for use in making tire treads which comprises terpolymerizing styrene, isoprene and 1,3-butadiene in an organic solvent at a temperature of no more than about 40.degree. C. in the presence of (a) at least one member selected from the group consisting of tripiperidino phosphine oxide and alkali metal alkoxides and (b) an organolithium compound.
U.S. Pat. No. 5,047,483 discloses a pneumatic tire having an outer circumferential tread where said tread is a sulfur-cured rubber composition comprised of, based on 100 parts by weight rubber (phr), (A) about 10 to about 90 parts by weight of a styrene, isoprene, butadiene terpolymer rubber (SIBR), and (B) about 70 to about 30 weight percent of at least one of cis 1,4-polyisoprene rubber and cis 1,4-polybutadiene rubber wherein said SIBR rubber is comprised of (1) about 10 to about 35 weight percent bound styrene, (2) about 30 to about 50 weight percent bound isoprene and (3) about 30 to about 40 weight percent bound butadiene and is characterized by having a single glass transition temperature (Tg) which is in the range of about -10.degree. C. to about -40.degree. C. and, further the said bound butadiene structure contains about 30 to about 40 percent 1,2-vinyl units, the said bound isoprene structure contains about 10 to about 30 percent 3,4-units, and the sum of the percent 1,2-vinyl units of the bound butadiene and the percent 3,4-units of the bound isoprene is in the range of about 40 to about 70 percent.
U.S. Pat. No. 5,272,220 discloses a styrene-isoprene-butadiene rubber which is particularly valuable for use in making truck tire treads which exhibit improved rolling resistance and tread wear characteristics, said rubber being comprised of repeat units which are derived from about 5 weight percent to about 20 weight percent styrene, from about 7 weight percent to about 35 weight percent isoprene, and from about 55 weight percent to about 88 weight percent 1,3-butadiene, wherein the repeat units derived from styrene, isoprene and 1,3-butadiene are in essentially random order, wherein from about 25 percent to about 40 percent of the repeat units derived from the 1,3-butadiene are of the cis-microstructure, wherein from about 40 percent to about 60 percent of the repeat units derived from the 1,3-butadiene are of the trans-microstructure, wherein from about 5 percent to about 25 percent of the repeat units derived from the 1,3-butadiene are of the vinyl-microstructure, wherein from about 75 percent to about 90 percent of the repeat units derived from the isoprene are of the 1,4-microstructure, wherein from about 10 percent to about 25 percent of the repeat units derived from the isoprene are of the 3,4-microstructure, wherein the rubber has a glass transition temperature which is within the range of about -90.degree. C. to about -70.degree. C., wherein the rubber has a number average molecular weight which is within the range of about 150,000 to about 400,000, wherein the rubber has a weight average molecular weight of about 300,000 to about 800,000, and wherein the rubber has an inhomogeneity which is within the range of about 0.5 to about 1.5.
U.S. Pat. No. 5,239,009 reveals a process for preparing a rubbery polymer which comprises: (a) polymerizing a conjugated diene monomer with a lithium initiator in the substantial absence of polar modifiers at a temperature which is within the range of about 5.degree. C. to about 100.degree. C. to produce a living polydiene segment having a number average molecular weight which is within the range of about 25,000 to about 350,000; and (b) utilizing the living polydiene segment to initiate the terpolymerization of 1,3-butadiene, isoprene and styrene, wherein the terpolymerization is conducted in the presence of at least one polar modifier at a temperature which is within the range of about 5.degree. C. to about 70.degree. C. to produce a final segment which is comprised of repeat units which are derived from 1,3-butadiene, isoprene and styrene, wherein the final segment has a number average molecular weight which is within the range of about 25,000 to about 350,000. The rubbery polymer made by this process is reported to be useful for improving the wet skid resistance and traction characteristics of tires without sacrificing tread wear or rolling resistance.
U.S. Pat. No. 5,061,765 discloses isoprene-butadiene copolymers having high vinyl contents which can reportedly be employed in building tires which have improved traction, rolling resistance and abrasion resistance. These high vinyl isoprene-butadiene rubbers are synthesized by copolymerizing 1,3-butadiene monomer and isoprene monomer in an organic solvent at a temperature which is within the range of about -10.degree. C. to about 100.degree. C. in the presence of a catalyst system which is comprised of (a) an organoiron compound, (b) an organoaluminum compound, (c) a chelating aromatic amine and (d) a protonic compound; wherein the molar ratio of the chelating amine to the organoiron compound is within the range of about 0.1:1 to about 1:1, wherein the molar ratio of the organoaluminum compound to the organoiron compound is within the range of about 5:1 to about 200:1, and herein the molar ratio of the protonic compound to the organoaluminum compound is within the range of about 0.001:1 to about 0.2:1.
U.S. Pat. No. 5,405,927 discloses an isoprene-butadiene rubber which is particularly valuable for use in making truck tire treads, said rubber being comprised of repeat units which are derived from about 20 weight percent to about 50 weight percent isoprene and from about 50 weight percent to about 80 weight percent 1,3-butadiene, wherein the repeat units derived from isoprene and 1,3-butadiene are in essentially random order, wherein from about 3 percent to about 10 percent of the repeat units in said rubber are 1,2-polybutadiene units, wherein from about 50 percent to about 70 percent of the repeat units in said rubber are 1,4-polybutadiene units, wherein from about 1 percent to about 4 percent of the repeat units in said rubber are 3,4-polyisoprene units, wherein from about 25 percent to about 40 percent of the repeat units in the polymer are 1,4-polyisoprene units, wherein the rubber has a glass transition temperature which is within the range of about -90.degree. C. to about -75.degree. C., and wherein the rubber has a Mooney viscosity which is within the range of about 55 to about 140.
U.S. patent application Ser. No. 08/524,666, filed on Sep. 8, 1995, discloses an isoprene-butadiene diblock rubber which has an excellent combination of properties for use in making automobile tire treads, said isoprene-butadiene diblock rubber being comprised of a butadiene block and an isoprene-butadiene block, wherein said butadiene block has a number average molecular weight which is within the range of about 25,000 to about 350,000, wherein said isoprene-butadiene block has a number average molecular weight which is within the range of about 25,000 to about 350,000, wherein said isoprene-butadiene diblock rubber has a glass transition temperature which is within the range of about -100.degree. C. to about -70.degree. C., wherein said isoprene-butadiene diblock rubber can optionally have a second glass transition temperature which is within the range of about -50.degree. C. to about 0.degree. C., wherein said isoprene-butadiene diblock polymer has a Mooney ML-4 viscosity at 100.degree. C. which is within the range of about 50 to about 140, and wherein the repeat units derived from isoprene and 1,3-butadiene in the isoprene-butadiene block are in essentially random order. U.S. patent application Ser. No. 08/524,666 further discloses tire tread compounds which are (1) blends of such isoprene-butadiene diblock rubbers with 3,4-polyisoprene rubbers, (2) blends of such isoprene-butadiene diblock rubbers with high cis-1,4-polybutadiene rubbers and (3) blends of such isoprene-butadiene diblock rubbers with natural rubber.


A mop for cleaning a floor includes a handle. A mop head is attached to the distal end of the handle. A sponge having a floor contact surface is removably attached to the mop head. A squeegee, having a floor wiping surface, is formed integral with a front surface of the mop head. The mop also includes a mechanism for wringing the sponge, coupled to the mop head.

Mops are floor cleaning implements which include a cleaning element, attached to an elongate handle, that contacts the floor during cleaning and that absorbs (and expels) liquid. Liquid remaining on the floor after mopping can be hazardous and also can reduce the attained cleanliness of the floor. As a result, many mop designs have been aimed at drying a floor surface after cleaning.
Some mops include absorbent sponges which act to absorb liquid remaining on the floor during the cleaning. Such designs have proved unsatisfactory in the level of dryness achieved. Wiping implements, such as those including squeegees attached to the end of a handle, also have been known to be used to wipe an area including liquid and then to separately use a mop including an absorption sponge for drying. Using two separate implements (mop and wiping), however, can be cumbersome. In addition, unless the mop and wiping implement are designed for complementary use, the level of dryness achieved often is unsatisfactory.
Prior art cleaning implements including both a wiping element (squeegee) and a sponge are known. To date, none of such dual element implements has proven to achieve satisfactory cleaning and drying while also being ergonomically easy to use. Some dual element implements include a squeegee and a sponge attached to different sides of the implement's handle, requiring the user to reverse the orientation of the implement to switch from using the sponge to using the squeegee and visa versa.
An example of a prior art cleaning implement including both a wiping element and cleaning element is described in U.S. Pat. No. 5,469,594 to Nolte. The cleaning implement described in Nolte includes a cleaning cloth and a scrapping element located on the same side, and attached to the distal end, of a handle. While the implement disclosed enables both cleaning and wiping without requiring a user to reverse the orientation of the implement, the implement suffers from a number of drawbacks. First, the Nolte implement is not well-suited for floor use due to the relative positions of the cleaning cloth, wiping element, and handle. It is designed specifically for cleaning vertically oriented surfaces, such as windows. Because the handle extends almost parallel to the bottom cleaning surface of the cloth, and because of the angle at which the wiping element extends from the implement, from an ergonomic standpoint, it would be extremely difficult to use the implement to clean floors. Additionally, due to the use of a cloth cleaning element having poor absorption capacity, and due also to the relative positions and spacing between the front of the cloth and the wiping element resulting in a limited wicking action, poor absorption by the cloth of the liquid being wiped results, leaving the floor wet after cleaning. The Nolte implement, because it is designed for cleaning a vertically oriented surface, relies in part on gravity for the liquid being wiped to reach and be absorbed by the cloth. It would be ergonomically difficult to use to clean a floor and would perform in an unsatisfactory manner.
In addition, to Applicant's knowledge, none of the prior art cleaning implements including both a wiping element and cleaning element, also includes a mechanism by which the cleaning element (i.e., sponge) may be wrung out.


1-Aminopyrrolidine which is obtained by reacting hydrazine hydrate with a compound represented by the following formula (I): EQU X.sup.1 --C.sub.4 H.sub.8 --X.sup.2 (I). wherein X.sup.1 and X.sup.2 are the same or different, and each represents a leaving group, in methanol; and a process for producing 1-aminopyrrolidine, comprising the step of reacting hydrazine hydrate with the above compound represented by formula (I) in methanol.

1. Field of the Invention
The present invention relates to a process for producing 1-aminopyrrolidine, and also to 1-aminopyrrolidine which is produced by the process.
2. Discussion of the Background
1-Aminopyrrolidine is a useful compound as a starting material or synthetic intermediate for tetrahydropyridazine derivatives used as herbicides. 1-Aminopyrrolidine is also a useful compound as a reducing agent, a deoxidizer, a scale inhibitor, an epoxy hardener, or the like.
With regard to the production process of 1-aminopyrrolidine, a process in which hydrazine hydrate is dissolved in isopropanol and then 1,4-dihalobutane and alkali are added thereto under heating is known (see SU 1525151-A1). However, the yield and purity of 1-aminopyrrolidine obtained by the process are not satisfactory.
Additionally, since isopropanol used in the process forms an azeotropic mixture with water in the reaction system, an anhydrous state is resulted in the reaction system during the reaction upon heating and, therefore, it is unavoidable that hydrazine hydrate used as a starting material changes to very toxic and dangerous anhydrous hydrazine whereby there is a problem in the operation.
Moreover, a small amount of isopropanol remains in the crude reaction product together with 1-aminopyrrolidine and fractionation of isopropanol from 1-aminopyrrolidine is very difficult whereby it is unavoidable that yield and purity of the desired product further decrease.

Process for producing 1-aminopyrrolidine, and 1-aminopyrrolidine according to the process


A surgical instrument is intended for use in cataract operations, and includes an ultrasonic probe in the form of a hollow needle which is connected with a suction conduit, through which lens debris is removed by suction together with a rinsing liquid. The rinsing liquid is supplied to the site of surgery via a flow path extending between the hollow needle and a sleeve surrounding this needle with a spacing from the needle. This sleeve extends approximately along the entire length of the hollow needle and has at its distal end at least one outlet aperture for the rinsing liquid. The sleeve has two layers, in which an outer jacket layer is made of tissue compatible plastic such as silicone. A small inner tube layer is positioned and fitted within the outer jacket. This inner tube is made of a material having greater stiffness than the material of the outer jacket, and radially supports the outer jacket.

1. Field of the Invention
The present invention relates to a surgical instrument for crushing crystalline eye lenses by means of ultrasound, and for removing the lens debris by suction. A hollow needle is connected with an ultrasound generator received in a handpiece. The lumen of the needle is communicatively connected in terms of flow with a suction duct extending through the handpiece. The needle is surrounded by a sleeve made of plastic, forming a flow path communicatively connected in terms of flow with a feed duct of the handpiece for feeding rinsing liquid. This sleeve extends approximately along the entire length of the hollow needle, and has near its front end at least one outlet aperture for the rinsing liquid.
2. The Prior Art
Instruments useful for eye surgery are described in DE 40 08 594 A. Publications of Hans Geuder GmbH of Hertzstrasse 4, Heidelberg, Germany, relate to these instruments also.
In connection with such instruments, a needle forming an ultrasonic probe extends axially from a handpiece and is detachably screwed into a needle receptacle. This receptacle is designed like a sleeve and is longitudinally movable within the handpiece at high frequency, in a known manner.
The needle is a hollow needle with a lumen for removing by suction the rinsing liquid, and the lens debris carried along by this liquid. The lumen is connected with a suction conduit through a duct extending through the handpiece. The rinsing liquid is fed through a ring duct surrounding the end of the needle receptacle that is screwed onto the hollow needle.
When the instrument is used as intended, a plastic cap is exchangeably screwed onto the end of the handpiece at the needle end. This cap receives in a suitably dimensioned chamber, the end of the needle receptacle and a head of the hollow needle screwed to the receptacle. The plastic cap has a sleeve extending from the chamber, and surrounds the needle approximately along its entire length, forming a ring duct which is sealed at the front end of the sleeve, and which has lateral outlet apertures for rinsing liquid.
When the instrument is used, the hollow needle with the sleeve of the plastic cap surrounding it is inserted through an incision at the edge of the cornea into the anterior chamber of the eye which is to undergo the surgery. In a known manner, the lens is crushed by means of the hollow needle forming the ultrasonic probe. This is because the probe, which has a beveled segment at its end in most cases, focuses ultrasound into the lens. Rinsing liquid flows through the ring duct surrounding the needle receptacle, which is longitudinally movable at high frequency. The rinsing liquid then flows into the rear chamber of the plastic cap receiving the front end of the needle receptacle and the head of the needle screwed to the receptacle. Then the rinsing liquid flows via the ring duct between the hollow needle and the sleeve of the plastic cap surrounding the hollow needle. In this manner, rinsing liquid is supplied to the site of the surgery, and is then removed again by suction from the anterior chamber of the eye together with lens debris, via the lumen extending through the hollow needle.
Instruments of this type have been used successfully and, as a rule, permit without any problems the removal of lenses in cataract surgery. This is done by crushing the lens by means of ultrasound into very fine particles, and then by removing these particles by suction together with the rinsing liquid previously admitted into the anterior chamber of the eye. At the same time, the rinsing liquid flowing through the ring duct serves as a thermal insulation of the sleeve against the hollow needle, which heats up highly during the course of an operation.
It has been found that the sleeve, which in most cases consists of flexible material such as silicone, can be easily manipulated within the area of the wound. However, the sleeve may be constricted by the pressure applied by the edges of the wound, until this sleeve comes to rest against the hollow needle. This may lead to local heating of the sleeve, and in extreme cases, it may even lead to strangulation of the ring duct. In connection with this ring duct strangulation, it may lead to overheating of the entire area between the site of the constriction and the needle point. This consequently leads to damage to the surrounding eye tissue caused by heat, and particularly to the edges of the wound enclosing the sleeve.
In order to avoid such damage, a hollow needle has been described in trade publication "CATARACT/IOL", which has radially outwardly projecting longitudinal ribs on which the sleeve surrounding the hollow needle supports itself radially if constrictions occur. With this state of the prior art, it is assured that feed ducts for the rinsing liquid are kept open with a high degree of probability. However, at the site of any constriction, the sleeve is in direct contact with the ribs of the hollow needle. Such contact areas may cause undesirable heat transfer from the hollow needle, which consists of metal, and which consequently has good thermal conductivity, to the sleeve, and to the eye tissue resting against the outside of the sleeve.

Surgical instrument for crushing crystalline eye lenses by means of ultrasound and for removing lens debris by suction


The operation for fastening the end portion of a cord-like body is facilitated, the operation is capable of being automated and the end of the cord-like body is capable of being secured in reliable fashion. A window for receiving a resilient cord retainer is formed near the outer circumference of at least one flange of flanges provided on both ends of a winding drum. The cord retainer comprises a central retaining finger, a pair of fixing fingers, one provided on either side of the central retaining finger, and a connecting piece interconnecting the retaining finger and fixing fingers. Distal ends of the fixing fingers and the connecting piece are abutted against upper and lower edges, respectively, of the window from outside the flange, and the distal end portions of the fixing fingers are crimped and secured by a folded-back portion on the outer circumference of the flange. The distal end portion of the central retaining finger lies on the inner side surface of the flange on the outer circumferential portion thereof, and the cord-like body is clamped and secured between the distal end portion of the retaining finger and the inner side surface of the flange at the outer circumferential portion thereof.

1. Field of the Invention
This invention relates to a reel for taking up cords and cord-like bodies, a wire clip used upon being attached to this reel, an apparatus, suited for use with this reel, for automatically taking up the cord-like body, and a device for fastening the end of the cord-like body.
The term "cord-like body" (or "cord") as used here is meant to cover all linear bodies (inclusive of standed wire), regardless of the material, referred to by various names such as steel cord, filament, metal wire, wire, rope, wire cable, synthetic resin line, steel and string.
2. Description of the Related Art
After a cord-like body, especially a strongly resilient cord-like body made of steel, is taken up on a reel, it is required that one end thereof be fastened to the reel at a suitable location. One structure for fastening the end of a cord-like body is described in the specification of Japanese Utility Model Application laid-Open No. 3-62069.
According to the structure disclosed, the outer circumferential portion of a reel flange is formed to have a slot from which the end of a cord is capable of being pulled, and the outside surface of the flange is provided with a fastening member, such as a clip or spring, at a position adjacent the slot. The cord end portion (a portion somewhat short of the end of the cord) is plastically deformed into a U- or C-shaped configuration, this portion is passed through the slot and is pulled out of the flange, and the curved portion that has been pulled out is fastened by being clipped.
With a structure of this type, the end portion of the cord must be pulled out through the slot, and fastening the cord end is a troublesome task. This structure does not lend itself to automation. Further, since the cord is bent or curved into a U- or V-shaped configuration, a fairly long portion of the cord is wasted.
Since a reel suited to automation of end fastening does not exist, it has not been possible, as a matter of course, to realize a device capable of automatically fastening the end of a cord and an apparatus which takes up the cord and includes the above-mentioned device.

Cord take-up reel, apparatus for automatically taking up the cord using said reel, and device for automatically fastening cord end


A method for removing components to be separated from industrial gases using an absorption and desorption processes having liquid absorbents. At least one absorption device and one desorption device are provided, at least a part of the laden solution leaving the absorption device is diverted before being heated and delivered to the head of the heat transfer section. The laden partial stream is heated by the steam rising from the lower part of the desorption device through heat exchange in the heat transfer section. The remaining stream of cold, laden solution leaving the absorption device is expanded by so the relief valve and the heat exchanger into a pressure relief vessel, such that the stream leaving the heat exchanger separates into a liquid and a gaseous state. The pressure in the pressure relief vessel pressure is lowered so that the total energy demand in absorption and desorption processes is reduced.

1. Field of the Invention
The present invention relates to an economical process for the removal of components to be separated from technical gases in absorption and desorption processes.
2. Description of the Related Art
Such technical gases are mostly natural gas or synthesis gas, the synthesis gas being generated from fossil raw materials such as crude oil or coals and from biological raw materials. Natural gas and synthesis gas contain useful valuable gases but also interfering components, such as sulphur compounds, in particular sulphur dioxide, carbon dioxide and other components to be separated such as hydrogen cyanide and water vapour. Beside natural gas and synthesis gas, flue gases from an incineration of fossil fuels are also included in the group of technical gases from which interfering components as, for example, carbon dioxide, are removed. The components to be separated may also be useful gases which are to be separated for a specific purpose.
Both physical and chemical absorbents can be used for absorption. Chemically acting absorbents are, for example, aqueous amine solutions, alkali salt solutions, etc. Selexol, propylene carbonate, N-methyl-pyrrolidone, morphysorb, methanol, etc. are physical absorbents.
It is known from prior art to remove components to be separated from technical gases in a circuit by means of absorption and desorption processes. The components to be separated are absorbed in the absorption device by the liquid absorbent. The gas which is insoluble in the solvent leaves the absorption device at the top, whereas the components to be separated remain in dissolved state in the liquid absorbent and leave the absorption device at the bottom. Before the laden solution is fed to the top of the desorption device for desorption, the laden solution is usually pre-heated by heat exchange with the hot, regenerated solution, by which part of the energy required for the desorption in the desorption device is recovered.
By means of a heating agent, a reboiler at the bottom of the desorption device serves to generate steam by partial evaporation of the solvent at the bottom inside the desorption device. Here, the generated steam serves as stripping agent to remove the components to be separated from the laden solution. The laden solution is freed by the stripping agent in countercurrent from the absorbed components to be separated. The stripped components to be separated leave the desorption device at the top, with the steam fraction of the stripping agent being condensed in a head condenser and returned to the desorption device. The regenerated solution which has been freed from the components to be separated leaves the desorption device at the bottom, with the solution usually being cooled after heat exchange has been carried out and returned to the top of the absorption device. This concludes the circuit of the absorption and the desorption process.
In the absorption, which in most cases is carried out at a working pressure of 1 to 100 bar, an absorption temperature of 20° C. to up to 70° C. has proved to be advantageous for removing the components to be separated from the technical gas.
The solution laden with the components to be separated can be regenerated by flashing to a lower pressure and/or stripping, the components to be separated being released again and/or stripped by means of steam. Upon completion of the regeneration process the absorbent can be cooled as required and used again.
The temperature required for desorption in a desorption device is higher than the temperature for absorption by means of the absorbent in an absorption device. The desorption device is usually operated at a temperature of 80° C. to 140° C. and an absolute pressure of 0.2 to up to 3 bar.
In absorption and desorption processes, heat recovery can be achieved in a heat exchanger by heat exchange between absorption solution to be heated and absorption solution to be cooled. This heat exchange serves on the one hand to pre-heat the fluid to be heated in a desirable manner, on the other hand the fluid to be cooled is cooled in a likewise desirable manner, so that the energy required for regeneration and supplied from external sources is significantly reduced.
Even in the case of an ideal heat exchange, in which the temperature approximation between the hot, regenerated solution and the heated laden solution is nearly zero, the absorption and desorption processes still require a lot of external energy for the regeneration of the solvent. For economical reasons, the heat exchanger is usually designed for a minimum temperature approximation of approx. 10 K between the hot, regenerated solution and the heated, laden solution. This results in an increase of the regeneration energy demand to be met by external sources.
EP 1 606 041 B1 discloses a method for the selective removal of sour-gas components from natural gas or synthesis gas, with the sour-gas components being removed selectively within two absorption stages by flashing the laden solution in two stages in a flash vessel to a selected pressure and then introducing it for desorption into the desorption device.
Another method for removing sour gas from natural gas is disclosed in DE 10 2005 030 028 A1 where the pressure of the laden solution between the absorption column and the stripping column is controlled stepwise in a way to ensure that as little additional energy as possible is required.
WO 2010/086039 A1 describes a method and a device for separating carbon dioxide from an exhaust gas of a fossil-fired power plant. The absorption and desorption process connected to the power plant combines the "split feed" and the "lean solvent flash" operating modes, in which a more favourable overall plant efficiency of the power plant process is only achieved if the two process stages are combined. The application of the method according to WO 2010/086039 A1 involves a significantly greater demand for equipment than in prior art, as it would here be necessary to install both a vacuum compression stage and another compression stage.
EP 1 736 231 A1 discloses a method and an apparatus for the removal of carbon dioxide, with several variants aiming at improving the energy efficiency being presented. On account of the thermal connection described, however, it is possible to recover only part of the energy which is supplied to the regeneration device, as most of the energy which is still contained in the exhaust steam from the flash vessel described in the disclosure is not recycled to the regeneration device but removed by an external cooler, thus being lost for the system. To ensure that the heat is transferred in an adequate manner from and to the enriched solution of the flash vessel, there is a significantly greater demand for equipment in form of additional heat exchangers, coolers, etc. A necessary intermediate feed in the absorber, for example, increases the required overall height of the absorber and thus also the cost.
On account of the increasing demand for resources an economical mode of operation in all fields has long become an important basis for the further development. The aim is therefore to provide an efficient and cost-saving process by which the energy demand of the whole plant is reduced.

Heat recovery in absorption and desorption processes


A chewing gum for mucosal delivery of cannabinoids, the chewing gum being formulated as an extruded chewing gum including water-soluble chewing gum ingredients mixed into water-insoluble gum base, wherein the gum base includes one or more natural resins in an amount of 10-40% by weight of the gum base, one or more elastomers in an amount of 3-30% by weight of the gum base, and one or more elastomer plasticizers in an amount of 8-50% by weight of the gum base, and wherein the chewing gum includes one or more cannabinoids, and wherein the release rate of the one or more cannabinoids is at least 10% by weight of the one or more cannabinoids within the first 5 minutes upon oral administration.

Cannabinoids are known for their health improving properties and have been used with respect to various medical purposes in the past. Among these medical purposes, cannabinoids have in particular been used for alleviating or treating different kinds of pain and counteracting side effects in relation to cancer treatment, such as nausea.
Traditionally, chewing gum has been described in the prior art as a vehicle for delivery of cannabinoids for alleviation or treatment of medical conditions, such as medical conditions associated with pain. While focus has generally been directed to clinical studies supporting various effects of cannabinoids to the human body, only limited attention is given in the prior art to improving such chewing gum formulations for the purpose of improving release of cannabinoids without addition of components that may interact with sensory properties of the gum.
Since chewing gum is a complex matrix of various types of components which all interact with each other, providing the chewing gum with specified properties, it appears that there is a need in the prior art for improved chewing gum platforms that offer a beneficial release of cannabinoids.
WO 2009/120080 discloses the use of chewing gum as a medical carrier and release vehicle of cannabinoids. The chewing gum disclosed herein may facilitate prolonged release of cannabinoids compared to other types of administering methods. However, various problems and challenges are associated with the chewing gum disclosed, partly based on the specific properties of cannabinoids. While certain specific conventional gum bases are used in formulating the chewing gum disclosed, the gum bases appear to have drawbacks for use in combination with cannabinoids.
In general, less attention is addressed in the prior art to the impact of the gum base and components in the gum base for the release properties of cannabinoids. Also, less attention is given on the impact of the gum base and components in the gum base for the sensory properties of chewing gum with cannabinoids. Here, important sensory properties include initial chew, texture, flavor perception, sweetness perception and off-notes associated with cannabinoids. These properties are both relevant from a convenience perspective in chewing gum, but certainly also in order to support an appropriate delivery of cannabinoids from chewing gum and avoid adverse side effects of cannabinoids.
Hence, there is a need in the prior art for improved chewing gum formulations that solve the above-referenced challenges and problems of the prior art. In particular, there is a need in the prior art for new gum base formulations for use in chewing gum that support appropriate delivery of cannabinoids combined with beneficial sensory properties.

This patent is subject to a terminal disclaimer.

Cannabinoid chewing gum with improved release of cannabinoids


Cake mixes utilizing unchlorinated wheat flour are disclosed comprising a quantity of unchlorinated wheat flour and about 20-40 bakers wt % unmodified starch. The cake mixes of this invention may further comprise 20-60 bakers wt % shortening, sweetening agents and a protein source. Also disclosed are cakes made from such mixes.

This invention relates to cakes and more particularly cake mixes made from unchlorinated wheat flour.
In the United States, cake flour is normally bleached with chlorine gas. Cakes made from chlorinated flour generally have more desirable volume, texture, grain and eating quality than cakes from unchlorinated wheat flour, Moreover, white cakes made from chlorinated flour have improved color. Despite the foregoing advantages, there is increasing pressure to discontinue the use of chlorinated flour due to health concerns. Many foreign countries prohibit the use of chlorinated flour in food products. A need thus exists for cakes made from unchlorinated flour that have volume and taste attributes similar to cakes made from chlorinated flour.


A management device transmits a response confirmation request to multiple image forming devices, receives a response from any one of the multiple image forming devices, and acquires data indicating an operational status of an image forming device by requesting the pertinent data from an image forming device that has issued a response.

Field of the Invention
The present invention relates to a technology which periodically collects data from an image forming device.
Description of the Related Art
Management devices have previously been proposed for acquiring and managing data such as operational information of image forming devices including printers or multifunction printers. When the image forming devices managed by management devices increase, the number of management devices also must increase. Therefore, from the standpoint of cost reduction and maintenance of management devices, it would be desirable to enable management of a greater number of image forming devices by a single management device. Moreover, in recent years, image forming devices have been proposed which economize on power consumption by setting the power to a power saving mode or by turning the power off at times of non-use in order to reduce emissions of greenhouse gases such as carbon dioxide. For example, Japanese Patent Application Laid-Open No. 2009-134584 discloses a technology in which an image forming device notifies a management device immediately before a power saving function is activated, and the management device that receives the notification collects job log information of the image forming device that is the management subject. According to this technology, a management device is able to efficiently conduct data collection with respect to an image forming device provided with a power status notification function.
However, although the management device disclosed in Japanese Patent Application Laid-Open No. 2009-134584 is able to efficiently conduct data collection with respect to an image forming device that is provided with a power status notification function, it is not able to efficiently conduct data collection with respect to an image forming device that is not provided with a power status notification function. For example, when the power of an image forming device that is not provided with a power status notification function is turned off, the management device is unable to detect it. As data acquisition cannot be carried out when data acquisition is attempted with respect to an image forming device whose power has been turned off, the management device enters standby until a timeout interval has elapsed. As a result, the time required for data collection is prolonged. Furthermore, when the number of image forming devices managed by a management device increases, a long period of time is required in order for the management device to conduct data collection from all the image forming devices. That is, the data collection interval is further prolonged. When the data collection interval is prolonged, the possibility increases that data loss may occur. This is because when the data volume of an image forming device of low data retention capacity reaches a maximum data volume, old data is deleted.

Management device, control method, and recording medium


A water-dilutable, microemulsion concentrate and pour-on formulations for water-insoluble insecticides such as amitraz is described, which includes a mixture of nonionic emulsifiers of defined amounts and HLB values, and, particularly, including a molecular sieve or a water stabilizer additive to provide a single phase, clear, physically and chemically stable system, without decomposition for an extended period of time.

1. Field of the Invention
This invention relates to a water-dilutable, microemulsion concentrate and pour-on formulations thereof, particularly for water-insoluble insecticides such as amitraz.
2. Description of the Prior Art
Many different delivery systems are available to deliver agriculturally active chemicals to an animal or plant site. However, such chemicals which are insoluble in water present a formidable challenge to effective use of such pesticide chemicals. Clearly, such chemicals cannot be readily formulated into an aqueous delivery system for pour-on application to animals or plants.
Accordingly, it is an object of this invention to provide a water-dilutable, microemulsion concentrate and pour-on formulations thereof for water-insoluble insecticides.
A specific object herein is to provide stable aqueous delivery systems for the water-insoluble amitraz insecticide.

Water-dilutable, microemulsion concentrate and pour-on formulations thereof


A method of scribing and separating chips on a semiconductor wafer (21) wherein the wafer (21) is patterned and a pattern of intersecting grooves is etched on a selected surface of the semiconductor wafer (21) in a pattern, preferably in a grid pattern, to define chip areas on that surface. Trenches (27) are then formed in the shape of the pattern and the selected surface is adhered to a tape (29). Light is then passed through the tape (29) and semiconductor wafer (21) from the pattern and a pattern of intersecting saw cuts or grooves (28) aligned with the light passing through the wafer (21) is formed. The saw cuts (28) extend from a surface of the wafer (21) opposing the selected surface and are aligned with the pattern of intersecting trenches (27).

1. Field of the Invention
This invention relates to a method and system for scribing and separating semiconductor chips formed on a semiconductor wafer.
2. Brief Description of the Prior Art
Chips formed on a single wafer of semiconductor material have generally been scribed and separated in the prior art by one of several available procedures. These prior art procedures generally provide for attachment of the wafer to an adhesive handling film with subsequent fracturing taking place along "streets" between rows and columns of chips to separate the chips. The chips are then removed from the handling film and processed in standard manner.
Fracturing of the wafer has generally been accomplished by (1) laser scribing a depression in the wafer along the "streets" and then breaking the wafer the rest of the way, (2) sawing a depression in the wafer along the "streets and then breaking the wafer the rest of the way, or (3) sawing entirely through the wafer along the "streets".
A problem arising from the above described prior art techniques has been the large amount of semiconductor material or chip surface area wasted due to the large "street" widths required in order to prevent damage to chips during chip separation. In the case of fracturing by the procedures listed above as (1) and (2), there is the possibility that the break in the wafer below the depression will not be straight, resulting in chips of non-uniform dimensions. Also, sawing can causes fractures at the wafer surface, this being a source of defects and decreased yields when sawing takes place at the surface containing the electrical components.

Trench scribe line for decreased chip spacing


The present invention relates to a composition comprising:    (a) a polymer having at least one repeating unit of formula

This invention relates generally to high silicon-content resin composition that can be used to form thin film thermosets, useful in forming low k dielectric constant materials and as well as hard mask and underlayer materials with anti-reflective properties for the photolithography industry.
As the dimensions of microelectronic devices become smaller, the importance of fully exploiting the physical properties of the materials used in their manufacture becomes more essential. This is particularly true of the dielectric materials that are used to insulate metal lines, vias and electronic components from one another because of the contributions to capacitance that these materials make. Silicon dioxide has been employed within the industry as a dielectric material for the manufacture of devices for nearly three decades, but may become less suitable in the future because of its relatively high dielectric constant (k˜4.1). Recently, however, siloxanes have made inroads into microlithography fabrication with low dielectric constant insulating layers and pattern transfer hard masks that exhibit antireflective properties.
Additionally, in current conventional semiconductor manufacturing, in order to prevent light reflection from being transmitted through the photo-resist, reflected off the substrate and back into the photoresist, where it can interfere with incoming light and so result in the uneven exposure of the photoresist, conventionally one or more antireflective layers may be deposited before the photoresist is deposited or spun on. In the absence of an antireflection coating, interference of reflected and incident exposure radiation can cause standing wave effects that distort the uniformity of the radiation through the thickness of the photoresist layer. Variations in reflectivity throughout the entire imaged areas can cause undesirable line width variation for features which are designed to have similar dimensions.

Abstract: where R1 is a non-hydrolysable group and n is an integer ranging from 1 to 3; and (b) a crosslinking catalyst. The composition is useful in forming low k dielectric constant materials and as well as hard mask and underlayer materials with anti-reflective properties for the photolithography industry.


A materials handling vehicle including a load handling assembly having a mast assembly, and a fork carriage assembly including a fork support and at least one fork assembly, the at least one fork assembly including a first fork member, which is fixed to the fork support, and a second fork member. The vehicle includes a tilt assembly that tilts the fork support relative to the mast assembly such that a central axis of the at least one fork assembly is positionable in a plurality of different positions relative to a horizontal direction. The vehicle includes a fork extension/retraction assembly that moves the second fork member relative to the first fork member in a first direction that is parallel to the central axis such that the fork extension/retraction assembly selectively moves the second fork member toward or away from the fork support in the first direction.

Known materials handling vehicles include a power unit, a mast assembly, and a platform assembly that includes a fork carriage assembly coupled to the mast assembly for vertical movement relative to the power unit. The mast assembly and platform assembly may each include components that are controlled by a hydraulic working fluid, such as pressurized oil. Valves provided within hydraulic fluid circuits associated with the mast and platform assemblies may control the flow of the working fluid to the components for effecting various functions performed by the components, such as raising/lowering, traversing (also known as side shifting), and tilting of the fork carriage assembly.

Materials handling vehicle having tilting fork carriage assembly with telescopic forks


A USB interface power supply method and power supply device for a time division multiple access load system reduce costs of using the capacitance while guaranteeing the performance. The USB interface power supply device includes: a USB interface, a soft-start circuit and a DC-DC direct current conversion circuit that are connected in sequence, wherein, an output of the DC-DC direct current conversion circuit supplies power to the time division multiple access load system. The power supply device also includes a capacitor, wherein: a first end of the capacitor is connected between the soft-start circuit and the DC-DC direct current conversion circuit, a second end of the capacitor is set to be grounded, and the capacitor is used for limiting an input current of the DC-DC direct current conversion circuit.

More and more types of USB data terminal products appear, with the promotion of data rate, the increase of functions and the increment of power consumption, a terminal with a USB interface has higher power supply requirements for the USB interface. According to the requirements of the USB communication protocol, the standard power supply current of the USB interface of the computer is 500 mA, in order to guarantee the security of interface power supply, a limited current circuit is set in all power supply circuits of the terminal products, the object is to protect that an output voltage of the USB power supply interface does not drop below a safe value, and there are basically two current-limiting ways, one is to perform current limiting at an output terminal of a Direct Current (DC)-DC converter, requirements of such current-limiting way for the output current ability of the DC-DC converter are not high, that is, the direct current converter with a low output current ability is appropriate when adopting this way; and the other one is to perform current limiting at an input terminal of the DC-DC converter, such current-limiting way is suitable for the output current ability being very high so that the current output by the converter individually can satisfy the demand of current when a time division multiple access load system bursts in working, and it is required to keep a working voltage of the input terminal of the converter not drop or drop a bit.
When the time division multiple access load system works in a burst mode, the instantaneous high current requires that a large capacitance with a certain capacity must be set in the terminal circuit to perform energy compensation, which makes voltages of a USB interface terminal and a load terminal not drop excessively.
In current-limiting ways of the input terminal of the traditional DC-DC converter, a voltage value of the USB protocol specification is not considered with regard to the selection of current-limiting components.

USB interface power supply method and power supply device for time division multiple access load system