Search PubMed⌕ Search

Biomedical subjects

E Hartmann

Publications and source records attributed to E Hartmann.

At least 55 records · Page 3Linked to original sources
PubMed1998

Blue light- and genetically-reversed gravitropic response in protonemata of the moss Ceratodon purpureus.

In darkness, protonemal filaments of Ceratodon purpureus (Brid.) grow negatively gravitropically (upwards). Red light induces a positive phototropic response mediated by the photoreceptor phytochrome. A red light treatment also has an inhibitory effect on the gravitropic response, an effect also mediated by phytochrome. In this study the effects of blue light on phototropism and on gravitropism were analysed. Unilateral blue light resulted in only a weak phototropic response, but markedly randomised growth direction. Blue light given together with a gravitropic stimulus reversed the gravitropism, changing it from negative to positive (filaments grow downward). The effect of blue light was also analysed with the mutant ptr116, which is defective in the biosynthesis of the phytochrome chromophore, and in a newly isolated mutant wwr2, which is positively gravitropic in darkness. Blue light induced the same reversal of gravitropism in ptrll6 as in the wild type, indicating that phytochrome is not involved in this process. In wwr2 the direction of gravitropism was unaltered by the blue light treatment. Light also affects chlorophyll content and the size of plastids, potential statoliths for gravitropism. Red light induced an increase in plastid size and chlorophyll content in the wild type but not in ptr116. Blue light induced a similar change in wild type plastids. It seems as though light-induced alterations of gravitropism are not simply mediated by alterations in plastid properties, and that red light and blue light evoke fundamentally different responses.

Bryopsida↗
PubMed1998

Similarities between the uterine decidual reaction and the "mesenchymal lesion" of the urinary bladder in aging mice.

The histopathologic characteristics of the decidual reaction in the uterus of aging mice and the "mesenchymal lesion/tumor" in the urinary bladder of aging mice are compared and found to be very similar. Both lesions consist of spindle and epithelioid cells, may contain round eosinophilic granules and possess nuclear progesterone receptors and cytoplasmic desmin. The decidual reaction derives from endometrial stromal cells, while the "mesenchymal lesion" apparently develops from mesenchymal cells near the trigone area, carrying or developing progesterone receptors. If the hypothesis is accepted that in aging mice the uterine decidual reaction and the "mesenchymal lesion" in the urinary bladder represent an equivalent type of tissue reaction, then it follows that the typical "mesenchymal lesion" is not a tumor and could be called more specifically "decidual-like reaction".

Aging↗
PubMed1998

Identification of a tRNA-specific nuclear export receptor.

In eukaryotes, tRNAs are synthesized in the nucleus and after several maturation steps exported to the cytoplasm. Here, we identify exportin-t as a specific mediator of tRNA export. It is a RanGTP-binding, importin beta-related factor with predominantly nuclear localization. It shuttles rapidly between nucleus and cytoplasm and interacts with nuclear pore complexes. Exportin-t binds tRNA directly and with high affinity. Its cellular concentration in Xenopus oocytes was found to be rate-limiting for export of all tRNAs tested, as judged by microinjection experiments. RanGTP regulates the substrate-exportin-t interaction such that tRNA can be preferentially bound in the nucleus and released in the cytoplasm.

Amino Acid Sequence↗
PubMed1998

Nightmare after trauma as paradigm for all dreams: a new approach to the nature and functions of dreaming.

Nightmares, far from being unsuccessful dreams or exceptions to rules about dreams, can be considered paradigms for all dreaming. They allow us to follow exactly how a disturbance or perturbation is handled by the processing systems in our minds. The data considered here consists of dream series in the weeks and months immediately following trauma in adults--in other words, nightmares and dreams occurring as the trauma resolves. It appears the traumatized person may dream first about the actual trauma (though not always), then, very quickly, the dreams appear to deal with the dominant emotion. Dreams of being overwhelmed by a tidal wave or being swept up by a whirlwind are common after almost any trauma. Clearly, such dreams are not about the sensory input from the actual trauma. Rather, the dreams are about the dominant emotion. The dreams contextualize (find a picture context for) the emotional concern. After trauma, the dominant emotion is usually first terror and fear, then often followed by guilt (such as survivor guilt). This too is pictured in the dream series. The same pattern of contextualizing an emotional concern can be seen in stressful situations, in pregnancy, or in patients whose lives are dominated by one emotion. This pattern is paradigmatic for all dreams, but it may be difficult to detect in "ordinary" dreams, because there may be a number of other relatively smaller emotional concerns present, as opposed to the one clear-cut dominant one (as after trauma). A theory of dreaming is sketched out based on these data which suggests that overall dreaming makes connections more broadly than waking in the nets of the mind, and that the connections are not made randomly but guided by the dreamer's emotional concerns. It is also suggested that the making of connections may be functional for the organism in the sense of "weaving in," or integrating, new material.

Adolescent↗
PubMed1998

Recombinant phytochrome of the moss Ceratodon purpureus: heterologous expression and kinetic analysis of Pr-->Pfr conversion.

The phytochrome-encoding gene Cerpu;PHY;2 (CP2) of the moss Ceratodon purpureus was heterologously expressed in Saccharomyces cerevisiae as a polyhistidine-tagged apoprotein and assembled with phytochromobilin (P phi B) and phycocyanobilin (PCB). Nickel-affinity chromatography yielded a protein fraction containing approximately 80% phytochrome. The holoproteins showed photoreversibility with both chromophores. Difference spectra gave maxima at 644/716 nm (red-absorbing phytochrome [Pr]/far-red-absorbing phytochrome [Pfr]) for the PCB adduct, and 659/724 nm for the P phi B-adduct, the latter in close agreement with values for phytochrome extracted from Ceratodon itself, implying that P phi B is the native chromophore in this moss species. Immunoblots stained with the antiphytochrome antibody APC1 showed that the recombinant phytochrome had the same molecular size as phytochrome from Ceratodon extracts. Further, the mobility of recombinant CP2 holophytochrome on native size-exclusion chromatography was similar to that of native oat phytochrome, implying that CP2 forms a dimer. Kinetics of absorbance changes during the Pr-->Pfr photoconversion of the PCB adduct, monitored between 620 and 740 nm in the microsecond range, revealed the rapid formation of a red-shifted intermediate (I700), decaying with a time constant of approximately 110 microseconds. This is similar to the behavior of phytochromes from higher plants when assembled with the same chromophore. When following the formation of the Pfr state, two major processes were identified (with time constants of 3 and 18 ms) that are followed by slow reactions in the range of 166 ms and 8 s, respectively, albeit with very small amplitudes.

Avena↗
PubMed1998

Fluorescence and photochemistry of recombinant phytochrome from the cyanobacterium Synechocystis.

Fluorescence and photochemical properties of phytochrome from the cyanobacterium Synechocystis were investigated in the temperature interval from 293 to 85 K. The apoprotein was obtained by overexpression in Escherichia coli and assembled to a holophytochrome with phycocyanobilin (PCB) and phytochromobilin (P phi B), Syn(PCB)phy and Syn(P phi B)phy, respectively. Its red-absorbing form, Pr, is characterized at 85 K by the emission and excitation maxima at 682 and 666 nm in Syn(PCB)phy and at 690 and 674 nm in Syn(P phi B)phy. At room temperature, the spectra are blue shifted by 5-10 nm. The fluorescence intensity dropped down by approximately 15-20-fold upon warming from 85 to 293 K and activation energy of the fluorescence decay was estimated to be ca 5.4 and 4.9 kJ mol-1 in Syn(PCB)phy and Syn(P phi B)phy, respectively. Phototransformation of Pr upon red illumination was observed at temperatures above 160-170 K in Syn(PCB)phy and above 140-150 K in Syn(P phi B)phy with a 2-3 nm shift of the emission spectrum to the blue and increase of the intensity of its shorter wavelength part. This was interpreted as a possible formation of the photoproduct of the meta-Ra type of the plant phytochrome. At ambient temperatures, the extent of the Pr phototransformation to the far-red-absorbing form, Pfr, was ca 0.7-0.75 and 0.85-0.9 for Syn(PCB)phy and Syn(P phi B)phy, respectively. Fluorescence of Pfr and of the photoproduct similar to lumi-R was not observed. With respect to the photochemical parameters, Syn(PCB)phy and Syn(P phi B)phy are similar to each other and also to a small fraction of phyA (phyA") and to phyB. The latter were shown to have low photochemical activity at low temperatures in contrast to the major phyA pool (phyA'), which is distinguished by the high extent (ca 50%) of Pr phototransformation at 85 K. These photochemical features are interpreted in terms of different activation barriers for the photoreaction in the Pr excited state.

Cyanobacteria↗
PubMed1997

Cloning of two novel human importin-alpha subunits and analysis of the expression pattern of the importin-alpha protein family.

The import of many proteins into the nucleus is mediated by the importin-alpha/beta heterodimer. While only one importin-beta gene has been found, several forms of importin-alpha have been described. In addition to the three human importin-alphas already identified, we report here the primary structure of two new human importin-alpha proteins. The five known human importin-alpha subunits can be classified into three subfamilies that appear conserved in higher eukaryotic organisms. We show by immunoblotting that the different importin-alpha subfamilies are expressed in a variety of human tissues and mammalian cell lines.

Amino Acid Sequence↗
PubMed1997

Characterization of recombinant phytochrome from the cyanobacterium Synechocystis.

The complete sequence of the Synechocystis chromosome has revealed a phytochrome-like sequence that yielded an authentic phytochrome when overexpressed in Escherichia coli. In this paper we describe this recombinant Synechocystis phytochrome in more detail. Islands of strong similarity to plant phytochromes were found throughout the cyanobacterial sequence whereas C-terminal homologies identify it as a likely sensory histidine kinase, a family to which plant phytochromes are related. An approximately 300 residue portion that is important for plant phytochrome function is missing from the Synechocystis sequence, immediately in front of the putative kinase region. The recombinant apoprotein is soluble and can easily be purified to homogeneity by affinity chromatography. Phycocyanobilin and similar tetrapyrroles are covalently attached within seconds, an autocatalytic process followed by slow conformational changes culminating in red-absorbing phytochrome formation. Spectral absorbance characteristics are remarkably similar to those of plant phytochromes, although the conformation of the chromophore is likely to be more helical in the Synechocystis phytochrome. According to size-exclusion chromatography the native recombinant apoproteins and holoproteins elute predominantly as 115- and 170-kDa species, respectively. Both tend to form dimers in vitro and aggregate under low salt conditions. Nevertheless, the purity and solubility of the recombinant gene product make it a most attractive model for molecular studies of phytochrome, including x-ray crystallography.

Apoproteins↗
PubMed1997

A novel class of RanGTP binding proteins.

The importin-alpha/beta complex and the GTPase Ran mediate nuclear import of proteins with a classical nuclear localization signal. Although Ran has been implicated also in a variety of other processes, such as cell cycle progression, a direct function of Ran has so far only been demonstrated for importin-mediated nuclear import. We have now identified an entire class of approximately 20 potential Ran targets that share a sequence motif related to the Ran-binding site of importin-beta. We have confirmed specific RanGTP binding for some of them, namely for two novel factors, RanBP7 and RanBP8, for CAS, Pse1p, and Msn5p, and for the cell cycle regulator Cse1p from Saccharomyces cerevisiae. We have studied RanBP7 in more detail. Similar to importin-beta, it prevents the activation of Ran's GTPase by RanGAP1 and inhibits nucleotide exchange on RanGTP. RanBP7 binds directly to nuclear pore complexes where it competes for binding sites with importin-beta, transportin, and apparently also with the mediators of mRNA and U snRNA export. Furthermore, we provide evidence for a Ran-dependent transport cycle of RanBP7 and demonstrate that RanBP7 can cross the nuclear envelope rapidly and in both directions. On the basis of these results, we propose that RanBP7 might represent a nuclear transport factor that carries an as yet unknown cargo, which could apply as well for this entire class of related RanGTP-binding proteins.

Amino Acid Sequence↗
PubMed1997

The yeast SPC22/23 homolog Spc3p is essential for signal peptidase activity.

In eucaryotic cells signal sequences of secretory and membrane proteins are cleaved by the signal peptidase complex during their transport into the lumen of the endoplasmic reticulum. The signal peptidase complex in yeast consists of four subunits. To date, three of these subunits have been functionally characterized. One of them, the Sec11p, is essential for viability of yeast cells. It shows significant homology to the mammalian SPC18 and SPC21 as well as to bacterial leader peptidases. Two other subunits, Spc1p and Spc2p, have been shown to be homologous to mammalian SPC12 and SPC25, respectively, and are not essential for protein translocation or signal peptide cleavage. We have purified and analyzed the fourth subunit of yeast signal peptidase, Spc3p. The protein is essential for viability of yeast cells. Depletion of SPC3 leads to accumulation of precursors of secretory proteins in vivo and to the loss of the signal peptidase activity in vitro. Therefore, in contrast to the bacterial leader peptidases, yeast signal peptidase requires a second subunit for its function.

Amino Acid Sequence↗
PubMed1997

Characterization of a peptide endothelium-derived constricting factor EDCF.

Endothelium regulates vascular tone by the release of dilator and constrictor mediators. Among the latter, besides endothelin, an 'endothelium derived constricting factor' EDCF sensitive to cyclooxygenase-inhibitors has been described. The aim of this study was to clarify the nature of this EDCF. Eluate from porcine aortic segments or supernatants (crude extracts) of porcine aortic segments were each tested for vascular effects in a bioassay system consisting of two endothelium-denuded acceptor vessels (rabbit abdominal aorta) before or after treatment with trypsin. The donor vessels were incubated with physiological saline solution with or without treatment with cycloheximide, quinacrine or indomethacine. Ultrafiltrates and fractions of a gelfiltration of the supernatants were also tested and compared with SDS-PAGE of these extracts. Finally, porcine aortic endothelial cells (PAEC) were cultured and the supernatant compared with that of the native aortae. A vasoconstrictive factor was released from the luminal surface of the porcine aortic segments, which if infused into the rabbit aortas induced two succeeding vasoconstrictions of 15-20 min duration each (the first 20 min after the first of extract-infusion, the second after 50 min) reaching 7% amplitude of a 0.2 mumol 1(-1) norepinephrine-induced constriction. These constrictions were enhanced if the crude extract of the porcine aortae was concentrated. This constricting factor was a protein with an approximative molecular weight of 9.000 Da. The release of this factor was insensitive to cycloheximide pretreatment indicating no de novo synthesis. However, the release of the factor could be markedly (50%) depressed by pretreatment with either quinacrine or indomethacine. The factor was not released from cultured PAEC. From these results, we conclude, that besides endothelin, endothelium luminally can release another endothelium-derived constricting factor named EDCF, a peptide with a molecular weight of 9.000 Da, which is not identical to endothelium and can induce long lasting vasoconstrictions. The release or synthesis of that EDCF seems to depend on cyclooxygenase and phospholipase A2 activity. We, thus, propose the name PLA2-sensitive EDCF for that factor.

Analysis of Variance↗
PubMed1997

Quantitation of alpha(2)-microglobulin after administration of structurally divergent chemical compounds.

alpha(2)-Microglobulin-induced nephropathy is a phenomenon which is exclusively found in adult male rats. Various chemicals are able to bind to alpha(2)-microglobulin thus inhibiting its proteolytic degradation in lysosomes of the P2 segment of the rat nephron. The accumulation of this protein in 'protein droplets' or 'hyaline droplets' leads to necrosis, followed by regeneration which possibly later results in the formation of tumours. Here we report the development of a monoclonal antibody which is specific for alpha(2)-microglobulin. It was utilized to measure alpha(2)-microglobulin concentrations in plasma and tissues, and to stain alpha(2)-microglobulin in fixed tissue slides. In two studies we administered to adult male Wistar rats two groups of compounds: (1) one group of structurally diverse compounds, which give an overview of chemical entities capable of inducing the accumulation of alpha(2)-microglobulin; and (2) another group of structurally closely related compounds (i.e. substituted benzene derivatives) for the purpose of elucidating possible structure-activity relationships. The degree of alpha(2)-microglobulin-induced nephropathy was determined by immunohistochemical staining of kidney sections. In addition, liver and kidney tissue and plasma concentrations of alpha(2)-microglobulin were not found to be elevated whereas kidney tissue concentrations were higher than the controls. The increase over control values ranged from 154% (1,4-dichloromethyl-benzene) to 321% [alpha-methyl-4-(1-methylethyl)-cyclohexanemethanol]. Comparing structurally related benzene derivatives, the hyaline droplet accumulating (HDA) potential was found to depend both on the type of substituent and its position at the aromatic ring. In general HDA activity increased in the order benzene approximately equal to phenol approximately equal to alkylated phenols < halogenated phenols < halogenated benzenes. Further QSAR studies are needed to provide a theoretical base for these observations.

Alpha-Globulins↗
PubMed1997

Receptor affinity purification of a lipid-binding adhesin from Haemophilus influenzae.

Thirteen clinical strains of Haemophilus influenzae, including types b, d, and untypeable, in vitro specifically recognize phosphatidylethanolamine (PE), gangliotetraosylceramide, gangliotriosylceramide (Gg3), sulfatoxygalactosylceramide, and to a lesser extent sulfatoxygalactosylglycerol. A PE affinity matrix was used to purify an adhesin of approximately 46 kDa from both type b and untypeable H. influenzae. This adhesin was a potent inhibitor of H. influenzae Gg3 and PE binding in vitro, and polyclonal antibodies specific for this protein prevented the attachment of H. influenzae Gg3 and PE and cultured HEp-2 epithelial cells in vitro.

Adhesins, Bacterial↗
PubMed1997

Phytochrome control of phototropism and chlorophyll accumulation in the apical cells of protonemal filaments of wildtype and an aphototropic mutant of the moss Ceratodon purpureus.

The aphototropic mutant line ptr116 of the moss Ceratodon purpureus shows characteristics of a deficiency in the phytochrome chromophore. Photoreversibility measurements indicate an approximately 20 time lower concentration of spectrally active phytochrome compared to wild-type, whereas normal phytochrome apoprotein levels are found on immunoblots. Feeding with the tetrapyrroles biliverdin, the proposed precursor of the phytochrome chromophore, or phycocyanobilin, which may replace the phytochrome chromophore, resulted in the rescue of ptr116 phototropism. The ptr116 mutant and the phenotypically-related mutant ptr1 contain lower chlorophyll levels than the wild-type. Chlorophyll content of wildtype and mutant tissue grown under different light conditions was estimated using conventional spectrophotometry of extracts and fluorimetrically, on single apical cells. Dark-grown tissue contained about 100 times less chlorophyll than tissue grown under standard white light conditions. Red light given for 24 h to dark adapted filaments induced an increase in the chlorophyll content in the wildtype, but not in ptr116. Blue light induced an increase in chlorophyll both in wildtype and in ptr116. The red light effect on the wildtype was partially reversible with far-red. If ptr116 was grown on phycocyanobilin, an increase in chlorophyll was also found when cells were irradiated with red light. The results indicate that phytochrome as well as a blue light photoreceptor regulate chlorophyll accumulation in C. purpureus protonemata. It can be assumed that in ptr116, the synthesis of the phytochrome chromophore is blocked specifically beyond the synthesis common to chlorophyll and the phytochrome chromophore and affects an enzymatic step between protoporphyrin and biliverdin.

Bryopsida↗
PubMed1997

Brief heat shock treatment induces a long-lasting alteration in the glycolipid receptor binding specificity and growth rate of Haemophilus influenzae.

After brief heat shock treatment, clinical strains of nontypeable Haemophilus influenzae show a long-lasting change in the binding specificity for glycolipids and a markedly increased growth rate in vitro. Non-heat-shocked H. influenzae specifically binds to phosphatidylethanolamine (PE), gangliotetraosylceramide (Gg4), and gangliotriosylceramide (Gg3) and binds minimally to sulfatoxygalactosylceramide (SGC; also called sulfatide). After a 5-min heat shock at 42 degrees C, strains of H. influenzae showed a marked increase in binding to SGC and acquired the ability to bind to sulfatoxygalactosylglycerol (SGG) in thin-layer chromatography overlays. Additionally, heat-shocked H. influenzae cells showed an increased growth rate (twofold). Increased sulfatide binding and growth rate were retained for approximately 60 generations, after which the heat-shocked organisms reverted to their original glycolipid binding pattern (i.e., PE, Gg3, and Gg4) and growth rate. Such organisms could then be reexposed to heat, and the heat shock phenotype would be reestablished. After exposure of the organisms to brief heat shock, Western blotting of a surface extract of H. influenzae with anti-bovine-brain hsp-70 monoclonal antibody showed an increase in two protein bands at 82 and 60 kDa. This antibody was a potent inhibitor of the binding of heat-shocked H. influenzae to SGC and SGG but had no effect on PE, Gg3, or Gg4 binding in vitro. In contrast, an antibody against an H. influenzae PE-Gg3-Gg4-binding adhesin that was recently identified (J. Busse, E. Hartmann, and C. A. Lingwood, J. Infect. Dis. 175:77-83, 1996) selectively inhibited the organism's binding to PE and Gg3. This indicates that cell surface hsp-70-related heat shock proteins can mediate H. influenzae attachment to sulfoglycolipids following heat shock. We suggest that such increased binding to sulfated glycolipids may be a response to fever following H. influenzae infection in humans.

Antibodies, Blocking↗