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T Rausch

Publications and source records attributed to T Rausch.

At least 19 recordsLinked to original sources
PubMed2001

In wounded sugar beet (Beta vulgaris L.) tap-root, hexose accumulation correlates with the induction of a vacuolar invertase isoform.

Wounding of sugar beet tap-root causes an induction of invertase activity, which contributes to post-harvest sucrose losses. In this first comprehensive monitoring of wound-induced invertase mRNAs, proteins, enzyme activities, and tissue hexose concentrations, the VI isoform responsible for wound-induced hexose accumulation in mature tap-root could be identified.

Adaptation, Physiological↗
PubMed1999

cDNA and genomic cloning of sugar beet V-type H+-ATPase subunit A and c isoforms: evidence for coordinate expression during plant development and coordinate induction in response to high salinity.

The plant V-type H+-ATPase (V-ATPase) does not only serve basic housekeeping functions but is also involved in stress-induced NaCl sequestration during salinity stress. To address the question whether the same isoforms conferring housekeeping functions are equally involved in the response to high salinity, we have isolated cDNA clones for subunits A and c, as representing the peripheral V1 complex and the membrane-integral V0 complex, respectively, from the halotolerant sugar beet (Beta vulgaris L., diploid variety). RNA blot analysis with gene-specific probes revealed a coordinate expression of the cloned subunit A and c isoforms during plant development and in response to high salinity. Also, in rapidly dividing suspension-cultured cells with 10-fold increased transcript amounts as compared to young leaf tissue, the ratio of transcripts for both genes was similar to the ratio found for transcripts in leaves of different age. We have then isolated partial genomic clones (BVA/70 for Beta V-ATPase 70 kDa subunit; BVA/16-1 for Beta V-ATPase 16 kDa subunit), including the promoter regions. Transcription start mapping revealed long 5'-UTR leader sequences (230 and 172 bases, respectively) for both genes. Both promoters contain putative G-box motifs in similar distance to the TATA boxes. For a quantitative comparison of relative promoter strength, the BVA/70 and BVA/16-1 promoters linked to the luciferase reporter gene (LUC) were delivered to sugar beet suspension-cultured cells by particle bombardment. The BVA/16-1 promoter showed a 1.7-fold higher activity as compared with the BVA/70 promoter. Salt treatment induced an increase of BVA/70 (+70%) and BVA/16-1 (+57%) promoter activities, concomitant with increased transcript amounts. The following sequences have been deposited at the EMBL database X98767: Beta vulgaris V-ATPase subunit A, cDNA clone; X98851, B. vulgaris V-ATPase subunit c isoform 1, cDNA clone; Y11038, B. vulgaris V-ATPase subunit A, partial genomic clone; Y11037, B. vulgaris V-ATPase subunit c isoform 1, partial genomic clone.

Amino Acid Sequence↗
PubMed1999

Cloning sulfur assimilation genes of Brassica juncea L.: cadmium differentially affects the expression of a putative low-affinity sulfate transporter and isoforms of ATP sulfurylase and APS reductase.

The heavy-metal accumulator Brassica juncea L. is a high-biomass crop able to extract heavy-metal ions from the soil, a substantial part being translocated from root to shoot. Previous work has shown that Cd accumulation is accompanied by massive formation of phytochelatins (PCs). Rapid de novo synthesis of PCs in roots and leaves requires an increased synthesis of the tripeptide glutathione (GSH), which in turn depends on increased sulfur assimilation. Therefore. we have cloned cDNAs for three enzymes involved in sulfur assimilation, i.e. a putative low-affinity sulfate transporter (LAST) and two isoforms each for ATP sulfurylase (ATPS) and APS reductase (APSR). As degradation of glucosinolates might provide an additional sulfur source under stress, we also cloned a myrosinase (MYR). RNA blot analysis of transcript amounts indicated that upon Cd exposure (25 microM) the expression of ATPS and APSR in roots and leaves of 6-week-old Brassica juncea plants was strongly increased, whereas the expression of MYR was unaffected. LAST transcripts were significantly reduced in the root but remained unchanged in the leaves. Concomitant with Cd induction of ATPS and APSR mRNAs, cysteine concentrations in roots and leaves increased by 81% and 25%, respectively, whereas GSH concentrations decreased in roots and leaves by 39% and 48%, respectively. In agreement with our previous report on Cd induction of gamma-glutamylcysteine synthetase in B. juncea, the results indicate coordinate changes of expression for several sulfur assimilation enzymes in response to an increased demand for cysteine during PC synthesis.

Amino Acid Sequence↗
PubMed1999

Ectopic expression of a tobacco invertase inhibitor homolog prevents cold-induced sweetening of potato tubers.

We have transformed potato with Nt-inhh cDNA, encoding a putative vacuolar homolog of a tobacco cell wall invertase inhibitor, under the control of the CaMV 35S promoter. In transgenic tubers, cold-induced hexose accumulation was reduced by up to 75%, without any effect on potato tuber yield. Processing quality of tubers was greatly improved without changing starch quantity or quality, an important prerequisite for the biotechnological use of Nt-inhh for potato transformation.

Amino Acid Sequence↗
PubMed1998

Effects of a partially hydrolyzed guar gum on intestinal absorption of carbohydrate, protein and fat: a double-blind controlled study in volunteers.

UNLABELLED: Some dietary fibres have been shown to affect the rate of absorption of dietary carbohydrate, protein and fat as well as exocrine pancreatic function. The aim of the study was to examine the effect of partially hydrolysed guar gum (BENEFIBER), on normal absorption of glucose, amino acid (arginine) and fat. In addition hepatic, pancreatic, renal and haematological side effects were evaluated. METHODS: A double blind, randomized and cross-over design was used. Each subject served as its own control. Ten healthy male volunteers participated in the study. Each subject was randomly assigned to two different dietary regimes for a period of 7 days each. The study was repeated with the other diet for another 7-day period after an interval of at least 1 week. In one study period the subjects took liquid formula diet without fibre and during the other study period they took the same diet with fibre. RESULTS: The results of the study demonstrated that BENEFIBER did apparently not interfere with the normal absorption of glucose, amino acid and fat. In keeping with these observations, insulin release and exocrine pancreatic function were not affected. Haematological, renal and hepatic toxicity were not observed in association with BENEFIBER consumption. CONCLUSION: We conclude that BENEFIBER is a safe source of soluble fibre.

Adult↗
PubMed1998

cDNA cloning and expression analysis of genes encoding GSH synthesis in roots of the heavy-metal accumulator Brassica juncea L.: evidence for Cd-induction of a putative mitochondrial gamma-glutamylcysteine synthetase isoform.

In roots of Brassica juncea L. cadmium (Cd) exposure (25 microM) induces a massive formation of phytochelatins (PCs), which is accompanied by an only moderate decrease (-20%) of the putative PC precursor glutathione (GSH). As PC formation in roots could be the result of local GSH de novo synthesis and/or depend on GSH import from the shoot, we have analyzed the expression of the enzymes involved in GSH synthesis in the root, namely OAS(thiol)lyase (OAS-TL; catalysing the last step in Cys biosynthesis), gamma-glutamylcysteine synthetase (gamma-ECS), and glutathione synthetase (GSHS). cDNA clones were isolated from a cDNA library prepared from heavy metal exposed roots. Protein sequences from cDNA clones encoding OAS-TL, gamma-ECS, and GSHS, all exhibited putative mitochondrial targeting sequences, however, for OAS-TL also two putative cytosolic isoforms were isolated. Furthermore, we have cloned several metallothionein cDNAs of the MT2 group. Northern blot analysis with coding region probes revealed that in roots of Cd-exposed plants transcript amounts for OAS-TL and GSHS were only moderately increased, whereas gamma-ECS mRNA showed a stronger increase. Expression analysis with 3'-UTR probes indicated that among the putative mitochondrial OAS-TL, gamma-ECS and GSHS isoforms only gamma-ECS was up-regulated in response to Cd exposure. Conversely, transcripts for MT2 appeared to be slightly reduced. The results indicate that in roots Cd-induced PC synthesis correlates with a moderate increase of expression of genes involved in GSH synthesis, the change for gamma-ECS being most pronounced.

Amino Acid Sequence↗
PubMed1998

Effect of transdermal application of nicotine on colonic transit in healthy nonsmoking volunteers.

The radio-opaque marker technique (ROMT) is a safe and noninvasive method to determine total colonic (TCTT) and segmental colonic transit times (SCTT). Previous results have shown that smoking volunteers had significantly longer TCTT than nonsmokers, but the underlying mechanism was not clear. We investigated the effect of transdermal nicotine application in two different doses in a non-blind randomized experiment involving three distinct phases. In phase 1 baseline transit times were determined with an abdominal X-ray after a 6-day period of marker ingestion and again after the following bowel movement to study the influence of a bowel movement just before the X-ray. TCTT was nearly twice as high before than after defaecation (42.6 h vs. 25.1 h, P < 0.05). The main acceleration was found in the rectosigmoid (RS) (18.6 h vs 7.1 h, P < 0.05) with no significant changes in right (RC) and left colon (LC). In phase 2 and 3 nicotine was applied in two doses of 17.5 mg day-1 and 35 mg day-1 in random order. Both doses resulted in a significant decrease of TCTT compared to the predefaecation baseline (42.6 h vs 32.2 h/28.2 h, respectively, P < 0.05). Again the main effect was located in the RS (18.6 h vs 9.9 h/7.6 h, P < 0.05). Short-term application nicotine results in a decrease of TCTT which is due to an accelerated transit in the RS.

Administration, Cutaneous↗
PubMed1998

Cloning of a tobacco apoplasmic invertase inhibitor. Proof of function of the recombinant protein and expression analysis during plant development.

Higher plants express several isoforms of vacuolar and cell wall invertases (CWI), some of which are inactivated by inhibitory proteins at certain stages of plant development. We have purified an apoplasmic inhibitor (INH) of tobacco (Nicotiana tabacum) CWI to homogeneity. Based on sequences from tryptic fragments, we have isolated a full-length INH-encoding cDNA clone (Nt-inh1) via a reverse transcriptase-polymerase chain reaction. Southern-blot analysis revealed that INH is encoded by a single- or low-copy gene. Comparison with expressed sequence tag clones from Arabidopsis thaliana and Citrus unshiu indicated the presence of Nt-inh1-related proteins in other plants. The recombinant Nt-inh1-encoded protein inhibits CWI from tobacco and Chenopodium rubrum suspension-cultured cells and vacuolar invertase from tomato (Lycopersicon esculentum) fruit, whereas yeast invertase is not affected. However, only in the homologous system is the inhibition modulated by the concentration of Suc as previously shown for INH isolated from tobacco cells. Highly specific binding of INH to CWI could be shown by affinity chromatography of a total cell wall protein fraction on immobilized recombinant Nt-inh1 protein. RNA-blot analysis of relative transcript ratios for Nt-inh1 and CWI in different parts of adult tobacco plants revealed that the expression of both proteins is not always coordinate.

Amino Acid Sequence↗
PubMed1997

In seedlings of the heavy metal accumulator Brassica juncea Cu2+ differentially affects transcript amounts for gamma-glutamylcysteine synthetase (gamma-ECS) and metallothionein (MT2).

Glutathione (GSH) is the precursor of the phytochelatins (PC), which in plants and fungi are involved in heavy metal sequestration. The regulatory enzyme gamma-glutamylcysteine synthetase (gamma-ECS) catalyzes the first step in GSH biosynthesis. For the heavy metal accumulator Brassica juncea L. a partial gamma-ECS cDNA was cloned by RT-PCR. Treatment of suspension-cultured dark grown seedlings with micromolar concentrations of CuSO4 resulted in a strong increase of gamma-ECS mRNA in roots and shoots, concomitant with an increase of GSH and phytochelatins. A significant up-regulation of gamma-ECS mRNA was observed at 25 microM CuSO4 (shoot growth: -11%), whereas maximum up-regulation was obtained at 100 microM CuSO4 (shoot growth: -60%). Unexpectedly, metallothionein 2 (MT2) mRNA was decreased in response to the CuSO4 treatments. CdSO4 at a concentration of 50 microM caused a 72% reduction in shoot growth without affecting the amounts of gamma-ECS- and MT2 mRNAs. ZnSO4 at a concentration of 500 microM did not reduce growth but induced transient increases of gamma-ECS- and MT2 mRNAs. The implications of the results with respect to differential regulation of gamma-ECS and MT2 during heavy metal exposure are discussed.

Arabidopsis↗
PubMed1997

Transport considerations for the critically ill child.

Transport of the critically ill child to and between hospitals has become a specialty field. Proper preparation and planning is critical on the part of the transferring party, the receiving facility, and the transport crew. Optimal patient outcome occurs when all parties involved understand each other and work together. Indications for transport, determination of transport mode, crew configuration of the transport team, necessary equipment, and physical examination including clinical interventions are key components of a successful transport. The following is a brief overview of these specific considerations involved when transporting the critically ill child.

Child↗
PubMed1997

[Symptomatic therapy of severe diarrhea in eosinophilic gastroenteritis with the somatostatin analog octreotide (Sandostatin)].

We describe a patient with eosinophilic gastroenteritis whose symptoms included severe watery diarrhea (1.5 l/d), abdominal cramps, and weight loss. Biopsies revealed massive eosinophilic inflammation of the entire gastrointestinal tract. The total plasma eosinophilic count was elevated by up to 30%. Infectious etiologies and chronic inflammatory bowel disease had been ruled out. By use of the somatostatin analogue octreotide the diarrhea decreased immediately in a dose-dependent fashion and the patient recovered within twelve days. In this case octreotide was helpful in controlling severe diarrhea and limiting the subsequent necessary i.v. fluid replacement.

Adult↗
PubMed1996

The 32 kDa tonoplast polypeptide Di associated with the V-type H+-ATPase of Mesembryanthemum crystallinum L. in the CAM state: A proteolytically processed subunit B?

In the facultative halophyte Mesembryanthemum crystallinum, the salt- or age-induced transition to crassulacean acid metabolism (CAM) leads to the occurrence of a tonoplast-bound 32 kDa polypeptide (Di). The alignment of its N-terminal protein sequence with protein sequences of recently cloned higher plant V-ATPase B-subunits indicates that Di may be derived from subunit B by proteolytic removal of a protein fragment of about 20 kDa from its N-terminus. Furthermore, an antiserum directed against Di cross-reacts with subunit B from Nicotiana tabacum. It inhibits both proton pumping and ATP hydrolysis of the holoenzyme in M. crystallinum. As Di remains firmly attached to the holoenzyme the proteolytic processing may have functional implications.

Adenosine Triphosphate↗
PubMed1996

Sucrose protects cell wall invertase but not vacuolar invertase against proteinaceous inhibitors.

Vacuolar (VI) and cell wall invertases (CWI) of higher plants can be inactivated in vitro and, possibly, in vivo by proteinaceous inhibitors. The respective mechanisms have not yet been compared. Therefore, partially purified CWI from transformed tobacco cells and VI from tomato fruit were preincubated with invertase-inhibitor fractions isolated from the same tissues. Both inhibitors were able to inhibit both invertases. However, VI was fully inhibited within less than 1 min by both inhibitors, whereas inactivation of CWI was much slower. Furthermore, CWI, but not VI, was strongly protected against inhibition by sucrose. A polyclonal antiserum directed against the tobacco inhibitor (I(NT)) cross-reacted with a 19 kDa polypeptide in the partially purified tomato inhibitor (I(LE)) fraction. The results indicate that I(NT) and I(LE)have similar structural properties, whereas the mechanism of inactivation is clearly different for CWI and VI.

Agrobacterium tumefaciens↗
PubMed1996

Down-regulation of plant V-type H+ -ATPase genes after light-induced inhibition of growth.

Cell extension growth in the mesocotyl tip of dark-grown Zea mays L. seedlings is dependent on vacuole enlargement and massive flux of ER and Golgi vesicles. Water flow into the expanding vacuole is driven by ion accumulation, which in turn is energized by the vacuolar H+-ATPase (V-ATPase). The V-ATPase energizes the secondary ion transport into the expanding vacuole. As light exposure leads to a strong inhibition of extension growth, the effect of light on transcript levels for subunits A and c of the V-ATPase was analyzed. Partial homologous cDNAs for subunit A and two isoforms of subunit c were cloned by RT-PCR. In dark-grown seedlings transcript levels for both subunits were much higher in the growing mesocotyl tip than in the fully differentiated mesocotyl tissue. Only in the tip region did light exposure lead to a strong and coordinate down-regulation of both mRNAs whereas in the differentiated mesocotyl only a slight decrease was observed. The results indicate that expression of the 'housekeeping' V-type H+-ATPase is strongly regulated in response to growth rate.

Adenosine Triphosphatases↗
PubMed1996

Salt stress induces an increased expression of V-type H(+)-ATPase in mature sugar beet leaves.

In the halotolerant sugar beet co-expression of V-ATPase and a vacuolar Na+/H(+)-antiporter provides a mechanism for vacuolar salt sequestration. To analyze salt-induced changes in the expression of the vacuolar H(+)-ATPase (V-ATPase) a partial cDNA of the proton-channel forming subunit c was cloned by RT-PCR. Southern blot analysis indicated a small gene family. In control plants transcript levels were high in roots and young growing leaves but low in fully expanded leaves. In mature leaves salt exposure (400 mM, 48 h) induced a strong increase in subunit c-mRNA. Transcripts for the catalytic subunit A followed a similar developmental and stress-modulated pattern, indicating a coordinate regulation of transcripts for both V-ATPase subunits. Concomittant with the mRNA increases the amount of V-ATPase protein increased as well.

Amino Acid Sequence↗
PubMed1996

Early salt stress effects on the differential expression of vacuolar H(+)-ATPase genes in roots and leaves of Mesembryanthemum crystallinum.

In Mesembryanthemum crystallinum, the salt stress-induced metabolic switch from C3 photosynthesis to Crassulacean acid metabolism is accompanied by major changes in gene expression. However, early effects of salt exposure (i.e. prior to Crassulacean acid metabolism induction) on genes coding for vacuolar transport functions have not yet been studied. Therefore, the expression of vacuolar H(+)-ATPase genes was analyzed in different organs of 4-week-old plants stressed with 400 mM NaCl for 3, 8, or 24 h. Partial cDNAs for the subunits A, B, and c were cloned and used as homologous probes for northern blot analysis. In control plants, the mRNA levels for the different subunits showed organ-specific differences. In fully expanded leaves, subunit c mRNA was very low but increased transiently during the light period. Plant organs also differed in their salt-stress response. In roots and young leaves, mRNA levels for all three subunits increased about 2-fold compared to control plants, whereas in fully expanded leaves only subunit c mRNA responded to salt. The results indicate that the expression of vacuolar H(+)-ATPase genes does not always involve a fixed stoichiometry of mRNAs for the different subunits and that the mRNA level for subunit c is particularly sensitive to developmental and environmental changes.

Amino Acid Sequence↗
PubMed1994

A 17-kDa Nicotiana tabacum cell-wall peptide acts as an in-vitro inhibitor of the cell-wall isoform of acid invertase.

When cell-wall invertase (CWI) from Nicotiana tabacum L. cell-suspension cultures, either non-transformed or transformed with Agrobacterium tumefaciens, was salt-eluted from intact cells and purified on Sulfopropyl-Sephadex (SPS) by pH-gradient elution, the enzyme lost about 50% of its activity during a 1-h incubation at pH 4.8. However, Western-blot analysis indicated no appreciable enzyme degradation. Re-chromatography of CWI peak fractions on SPS using NaCl-gradient elution showed the presence of a 17-kDa peptide (p17) in fractions with low CWI activity but strong CWI immunosignal (Weil and Rausch 1994, Planta 193, 430-437). When separating CWI from p17 by Concanavalin A (Con A)-Sepharose chromatography, inhibition could be restored by incubating the inhibitor-containing fraction with inhibitor-free CWI. More than 90% of CWI could be inhibited, suggesting that all CWI was susceptible to p17 binding. The presence of divalent metal ions (Ca2+, Mg2+, Zn2+) during pre-incubation of CWI with p17 reduced CWI inhibition substantially. Also, sucrose protected CWI against inhibition by p17 (half-maximum protection at 1.3 mM). Binding of p17 to CWI during a 1-h pre-incubation was pH-dependent, pH 4.5 causing maximum inhibition, whereas at pH 6.5 no inhibition was observed. Gel-permeation chromatography revealed that the native inhibitor acts as a monomer. Immunoprecipitation of CWI co-precipitated p17, confirming direct binding of p17 to CWI. When fractions containing CWI and p17 were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent Western blotting a diffuse immunosignal of 86-90 kDa was observed (in addition to the prominent CWI signal at 69 kDa).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗