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R Landgraf

Publications and source records attributed to R Landgraf.

At least 181 records · Page 10Linked to original sources
PubMed1994

Lack of fever suppression or central AVP release in 1K1C hypertensive rats.

Previous studies from our laboratory showed a transient suppression of the febrile response to intracerebroventricular (i.c.v.) PGE1 in the one-kidney, one-clip (1K1C) model of hypertension. This may have been due to an enhanced vasopressinergic transmission since arginine vasopressin (AVP), acting within the central nervous system (CNS), is thought to mediate endogenous antipyresis. These initial experiments utilized a protocol for the induction of 1K1C hypertension which produced an initial rapid rise in blood pressure, evident by day 4 following surgery, with a corresponding inhibition of the febrile response. The present experiments utilized a more slowly developing 1K1C hypertension (evident by day 12 following surgery) to firstly attempt to determine if inhibition of the febrile response is due to the actual change in blood pressure or to neural signals arising from the clipped kidney, and secondly to determine if the concentration of AVP in push-pull perfusates of the ventral septal area (VSA) of pyrogen-treated sham-operated and 1K1C rats were altered. In urethane-anaesthetized rats, i.c.v. PGE2 evoked brisk monophasic fevers in both 1K1C and sham-operated animals, with no significant difference between fever heights. Consistent with this, we found no increase in immunoreactive AVP from perfusates of the VSA of 1K1C rats. These results suggest that there is no inhibition of the febrile response to PGE2 when a slower developing hypertension is induced, nor is there an elevated release of AVP into the VSA under our conditions. We conclude that a rapid increase in blood pressure, and not high blood pressure per se, is required to produce an inhibition of the febrile response.

Alprostadil↗
PubMed1994

Oligonucleotide-directed nucleic acid scission by micrococcal nuclease.

"Sequence-dictated" scission of DNA can be achieved by tethering a fusion protein composed of glutathione S-transferase and attenuated micrococcal nuclease (MN) to a targeting oligonucleotide using Cibacron blue (CB) F3G-A. Deoxyoligonucleotides derivatized with this dye bind to the fusion protein in gel mobility shift assays. This binding scheme was successfully used to achieve site-specific scission of a single-stranded DNA substrate after hybridization with a CB-derivatized complementary oligonucleotide. Although covalently cross-linked hybrids of MN and oligonucleotides have been successfully used in the past to target nucleolytic activity, this novel scheme opens new possibilities for targeting and probing both DNA and RNA sequences by allowing the addition of the nuclease subsequent to hybridization.

Base Sequence↗
PubMed1994

Direct osmotic stimulation of the hypothalamic paraventricular nucleus by microdialysis induces excessive grooming in the rat.

This study examined the effects of direct osmotic stimulation of the hypothalamic paraventricular nucleus (PVN) on grooming behavior of adult male rats. Animals chronically fitted with a microdialysis probe in the right PVN responded to dialysis with hypertonic (containing 1 M NaCl) artificial cerebrospinal fluid (aCSF) with grooming behavior (to approx. 500%, P < 0.01, ANOVA) and a significantly reduced time resting (to approx. 20%, P < 0.01, ANOVA), compared with controls which received either the same treatment in the right supraoptic nucleus or in which the PVN was dialyzed with isotonic (0.15 M NaCl) a CSF. These results indicate that direct osmotic stimulation of the PVN via microdialysis is able to trigger and to maintain excessive grooming activity in rats; the novel experimental approach used here provides the potential to investigate the involvement of endogenous, intracerebrally released substances in the grooming response.

Animals↗
PubMed1994

Vasopressin facilitates its own release within the rat supraoptic nucleus in vivo.

A combined microdialysis/microinfusion technique was used to investigate whether arginine vasopressin (AVP) is involved in the regulation of its own release into the extracellular fluid of the supraoptic nucleus in vivo. While intranuclear neuropeptide release was monitored, 0.33 microliter of either vehicle, lysine vasopressin (LVP; 10 ng microliters -1) or a V1/V2 AVP receptor antagonist (100 ng microliters -1) was infused into the supraoptic nucleus of adult male Wistar rats before and during direct osmotic stimulation of the nucleus via the microdialysis probe. Administration of LVP increased basal AVP release, whereas administration of the V1/V2 receptor antagonist attenuated the increase in AVP release during osmotic stimulation observed in vehicle-treated controls. Taken together, these results indicate a receptor-mediated positive feedback action of endogenous AVP on its own release within the supraoptic nucleus.

Animals↗
PubMed1994

[Technics and results of pancreas transplantation].

Long-term normoglycaemia cannot be achieved in patients with insulin dependent diabetes mellitus neither with conventional nor with intensified insulin therapy. The only ideal method to obtain this seems the islet cell or pancreas transplantation. The number of pancreas transplantation approaches 5000 all over the world. The first simultaneous pancreas-kidney transplantation in Germany was performed in 1979 by the Munich group. Till 1991 in Grosshadern 141 pancreas transplantations have been performed. At the beginning duct occlusion (n = 106) later bladder drainage (n = 35) were used as a standard procedure. The authors discuss in detail the indications and contraindications, the types of pancreas transplantation, the different diversions of exocrine secretion. They analyse the effect of pancreas transplantation upon diabetic metabolism, retinopathy, neuropathy, nephropathy and quality of life, based on own experiences and literary data. At present the indication for pancreas transplantation is the stadium of late complications in IDDM. Because of the definitive lesions its beneficial effect is limited. After successful transplantation the peripheral (and autonomic?) neuropathy improves, the retinopathy seems to remain stabile, and the pancreas protects the transplanted kidney against recurrent diabetic nephropathy. Most patients will become insulin independent with tight metabolic control, but the complications of immunosuppressive therapy must be taken into consideration. The working ability and the quality of life seem to improve considerably.

Contraindications↗
PubMed1994

Structure of the Escherichia coli Fis-DNA complex probed by protein conjugated with 1,10-phenanthroline copper(I) complex.

The Escherichia coli Fis (factor for inversion stimulation) protein functions in many diverse biological systems including recombination, transcription, and DNA replication. Although Fis is a site-specific DNA-binding protein, it lacks a well-defined consensus recognition sequence. The electrophoretic mobility of Fis-DNA complexes, along with considerations of the Fis crystal structure, indicates that significant deformation of DNA occurs upon Fis binding. To investigate the structure of Fis-DNA complexes, the chemical nuclease 1,10-phenanthroline-copper complex (OP-Cu) has been linked to four specific sites within the Fis DNA-binding domain. Two of these Fis-OP derivatives were active in cleaving DNA. The scission patterns obtained on four different Fis binding sites indicate that Fis positions itself on these highly divergent DNA sequences in a very similar fashion. The patterns of cleavage of a derivative at Asn-98 generally support a model of a Fis-DNA complex that contains specific bends within the core-recognition sequence. Data from a second Fis-OP derivative at Asn-73 provides evidence for greater wrapping of flanking DNA around the sides of the Fis protein than was previously postulated. The cleavage efficiency of flanking segments varies, suggesting that the extent of DNA wrapping is sequence dependent. Specific amino acids on Fis are implicated in promoting this DNA wrapping.

Base Sequence↗
PubMed1994

Corticotropin-releasing hormone (CRH) antisense oligodeoxynucleotide treatment attenuates social defeat-induced anxiety in rats.

1. The neuropeptide corticotropin-releasing hormone (CRH) is the main mediator of the neuroendocrine and behavioral response to stress. End-capped phosphorothioate antisense and sense oligodeoxynucleotides (ODN) corresponding to the start coding region of rat CRH mRNA were infused intracerebroventricularly (30 micrograms/3 microliters per injection) three times at 12 hr intervals. Six hours after the last injection rats were subjected to social defeat stress and subsequently tested on the elevated plus maze. 2. Socially defeated CRH antisense-treated rats displayed markedly reduced anxiety-related behavior, as they spent significantly more time in the open arms of the plus maze compared to sense ODN- and vehicle-treated animals. 3. In controls, social defeat evoked a stress-induced elevation of CRH mRNA and CRH in the hypothalamus and a significant increase in plasma corticotropin (ACTH) levels. These parameters were attenuated in antisense-injected rats. 4. Our results suggest that CRH antisense treatment is effectively suppressing the neuroendocrine and behavioral effects of social defeat.

Agonistic Behavior↗
PubMed1994

Microdialysis administration of vasopressin into the septum improves social recognition in Brattleboro rats.

The role of septal arginine vasopressin (AVP) in a social recognition test was investigated in both homozygous Brattleboro (HO-DI) and normal Long-Evans rats. To do this, the duration of investigation of conspecific juveniles by untreated adult males of both rat strains was measured before and after inter exposure intervals of 30 and 120 min. Additionally, a microdialysis administration technique was used to administer synthetic AVP (0.2 or 2.0 ng) or its V1 receptor antagonist d(CH2)5Tyr(Me)AVP (5.0 ng) into the mediolateral septum concomitantly with the behavioral test. Untreated HO-DI rats showed an impaired social recognition compared with untreated Long-Evans rats. A similarly impaired performance was observed after V1 receptor antagonist treatment of Long-Evans rats. Microdialysis administration of synthetic AVP, on the other hand, significantly improved social recognition in both rat strains. The data suggest that endogenous AVP in the septal brain area is critically involved in the acquisition, storage, and/or recall of olfactory cues in rats.

Animals↗
PubMed1994

Opioids influence neurohypophysial but not central oxytocin release following direct hyperosmotic stimulation of the supraoptic nucleus in urethane-anaesthetised rats.

Microdialysis was used to apply an osmotic stimulus (0.5 M NaCl-aCSF) into both supraoptic nuclei (SON) to investigate the role of endogenous opioid peptides in the control of both central and peripheral oxytocin release in response to this stimulus. There were no differences in central peptide release during direct hyperosmotic stimulation between groups of rats given either vehicle, morphine (5 mg/kg) or naloxone (5 mg/kg) intravenously. Naloxone potentiated oxytocin release into blood; this suggests that endogenous opioid peptides at the level of the neurohypophysis, but not in the SON are important modulators of oxytocin release to this stimulus. However morphine blocked oxytocin release into blood indicative of a central inhibitory action on the firing rate of oxytocin neurones, contrasted with insensitivity to morphine of oxytocin secretion from the dendrites stimulated directly by hyperosmolarity.

Animals↗
PubMed1994

Crosstalk in the magnocellular system during osmotic stimulation of one supraoptic nucleus.

Neural connections linking the four magnocellular nuclei, i.e., the paired supraoptic (SON) and paraventricular (PVN) nuclei, may contribute to the simultaneous and parallel changes in firing patterns of oxytocinergic neurons during reflex milk ejection. To investigate these neural connections in the absence of suckling, intranuclear release of oxytocin (OT) was stimulated by microdialysis of hypertonic CSF containing 1 M NaCl (HS-CSF) into the right SON area and glucose metabolism of both SONs and PVNs and the neural lobe of virgin and lactating (10-12 day) rats was mapped by the autoradiographic [14C]deoxyglucose (DG) method. OT in the microdialysates and in plasma, obtained before and after 80-90 min of dialysis with CSF or HS-CSF, was quantified by RIA. In both virgin and lactating rats, microdialysis of HS-CSF unilaterally into the SON area significantly (p < 0.05) increased release of OT in the nucleus and into plasma, which was associated with enhanced (p < 0.05) metabolic activity in the ipsilateral and contralateral SON and the neural lobe but not in either PVN. Compared with virgins, lactating rats were less active, had lower (p < 0.05) glucose utilization in the hypothalamo-neurohypophysial system, and less (p < 0.05) OT in plasma during microdialysis of HS-CSF into the SON area. The osmotic stimulus did not activate neural structures (suprachiasmatic and medial amygdaloid nuclei) near the SON in either hemisphere. Thus, neural connections or, less likely, transport of OT via the subarachnoid space, may function to recruit activation of cells in the contralateral SON following hypertonic stimulation of cells in the other SON.

Animals↗
PubMed1994

Vasopressin antisense oligonucleotide induces temporary diabetes insipidus in rats.

The purpose of this study was to downregulate the transcriptional message of arginine vasopressin (AVP) by antisense treatment. A complete phosphorothioate antisense oligodesoxynucleotide corresponding to the beginning of the coding region of rat AVP mRNA was constructed and injected into the lateral ventricle of rats. Within 3-6 h animals exhibited a temporary diabetes insipidus, which lasted up to 9 h. Accordingly, vasopressin immunoreactivity in the hypothalamic nuclei was reduced. Our results demonstrate that a specific and reversible inhibition of neuropeptide expression can be accomplished in the intact hypothalamo-neurohypophysial system by antisense treatment, thus providing a novel tool for studies on stimulus-secretion coupling in vivo.

Animals↗
PubMed1994

Systemic osmotic stimulation increases vasopressin and oxytocin release within the supraoptic nucleus.

Vasopressin (VP) and oxytocin (OT) are released within the hypothalamic nuclear region in response to direct microdialysis with hypertonic solutions. Experiments were performed to determine whether systemic osmotic stimulation causes changes in intranuclear peptide release within the supraoptic nucleus (SON). A hypertonic sodium chloride solution was injected intraperitoneally (i.p.) or intravenously (i.v.) and microdialysis techniques were used to simultaneously monitor central and peripheral peptide release in urethane anesthetized rats. Systemic osmotic stimuli elicited increases in intranuclear peptide release which were delayed and long-lasting, occurring over a 2.5 h period. In contrast, plasma peptide levels peaked at 30-min after the stimulus. The results demonstrate that increased plasma sodium elicits an increase in VP and OT release into the extracellular space of the hypothalamic SON. The different patterns of peptide release in plasma and brain point toward the possibility of independently regulated release into the different compartments.

Animals↗
PubMed1994

DNA-binding proteins as site-specific nucleases.

DNA-binding proteins can be converted into site-specific nucleases by linking them to the chemical nuclease 1,10-phenanthroline-copper. This can be readily accomplished by converting a minor groove-proximal amino acid to a cysteine residue using site-directed mutagenesis and then chemically modifying the sulphydryl group with 5-iodoacetamido-1,10-phenanthroline-copper. These chimeric scission reagents can be used as rare cutters to analyse chromosomal DNA, to test predictions based on high-resolution nuclear magnetic resonance and X-ray crystal structures, and to locate binding sites of proteins within genomes.

Catalysis↗
PubMed1994

Long-term follow-up of lipid metabolism and rheologic properties after successful pancreas and kidney transplantation.

The long-term effect of pancreatic and kidney transplantation (spkt) on blood viscosity, lipid metabolism and skin microcirculation in insulin-dependent diabetes mellitus (IDDM) was studied because impaired rheological properties of blood may play a role in the development of diabetic micro- and macroangiopathy. 46 IDDM-patients (16 f/30 m; 23 +/- 34 y mean duration of diabetes; 60 +/- 14 mos mean follow up period) underwent spkt (Gr.I: n = 28) or solitary kidney (Gr.II: n = 18) transplantation, and were compared with healthy controls (C). Rheological measurements were performed with Mooney-Ewart rotation-viscosimeter determining whole blood viscosity (WBV), at shear rates 1, 5, 10, 20, 50, 100, 200 sec(-1). Triglycerides, total and HDL-, LDL- and VLDL cholesterol and fibrinogen were measured. Microcirculation was estimated by transcutaneous oxygen tension measurement (tcpO2) and laser speckle method, in the forefoot area. Hemoglobin A1 was normalized only in group I (I: 7.2 +/- 0.2%; II: 8.3 +/- 0.3%; C: < 8%). WBV at low shear (1, 5, 10) was increased in both groups, when compared to healthy controls (I: 12.4 +/- 2; 12.5 +/- 1; 6.8 +/- 0.5 mpas; II: 18.7 +/- 2; 13.4 +/- 15; 9.4 +/- 1 mpas; C: 7.5 +/- 0.5; 6.7 +/- 0.3; 5.4 +/- 0.2 mpas; P < 0.05). Plasma fibrinogen was elevated in both groups compared to normals: (I: 384 +/- 19; II: 448 +/- 20; C: 250 +/- 50 mg/dl; P < 0.05). There was a positive influence of spkt on skin microcirculation: tcpO2/prior tx: I: 44 +/- 3; II: 49 +/- 6 mmHg; post tx: I: 59 +/- 4; II: 42 +/- 3 mmHg. Laser speckle prior tx I: 3.3 +/- 0.3; II: 4.7 +/- 0.2 rel. U.; post tx: 3.8 +/- 0.2; II: 4.3 +/- 0.2 rel. U. Patients with progression of angiopathy showed still higher fibrinogen and shear rates (P < 0.05). There was no significant difference for total HDL-, LDL- and VLDL cholesterol. Despite normalization of glucose metabolism and significant improvement of microcirculation in spkt patients, fibrinogen and the shear rates are increased indicating a persisting "individual" vascular risk. It is suggested that an additional hemorheological approach in the treatment posttrransplant might prevent the progression of vascular complications.

Adult↗
PubMed1994

Osmotic stimulation of the supraoptic nucleus: central and peripheral vasopressin release and blood pressure.

Experiments were performed to determine the effect of direct osmotic stimulation of the supraoptic nucleus (SON) on central and peripheral vasopressin (AVP) release and arterial pressure. A microdialysis method was used to deliver hyperosmotic NaCl, mannitol or urea bilaterally into the SON and to sample SON extracellular fluid and blood. Simultaneous brain and blood microdialysis showed that hyperosmotic NaCl increased central and peripheral AVP release and increased mean arterial pressure (MAP). The pressor response was not blocked by intravenous injection of a V1-receptor antagonist, D(CH2)5Tyr(Me)AVP, suggesting that circulating AVP was not involved in that response. Hyperosmotic mannitol or urea caused an increase in central peptide release, but failed to affect MAP or peripheral AVP release. The results suggest that central AVP release within the SON may be due to osmoreceptor stimulation while the peripheral effects on AVP release and MAP are specific for sodium. The results also demonstrate the utility of brain and blood microdialysis for the delivery of stimuli into specific brain regions with simultaneous monitoring of central and peripheral peptide release.

Animals↗
PubMed1994

Reduced febrile responses to pyrogens after lesions of the hypothalamic paraventricular nucleus.

The hypothalamic paraventricular nucleus (PVN) is recognized as a major site of autonomic control, but the role of this nucleus in thermoregulation is unclear. Therefore the role of the PVN in the febrile response and in the maintenance of normal body temperature was investigated. Conscious, unrestrained rats with chronic lesions of the PVN received intracerebroventricular injections of several doses of prostaglandin (PG) E2 or intraperitoneal applications of Escherichia coli lipopolysaccharide. The body temperatures of both lesioned and sham-operated animals, monitored via radio telemetry, were compared. Intracerebroventricular PGE2 at doses of 10, 25, and 50 ng caused dose-dependent fevers in both PVN-lesioned and sham-operated animals, which at lower doses were smaller in the lesioned animals than in the sham-operated animals. Intraperitoneal lipopolysaccharide application, 50 micrograms/kg body wt, evoked a significantly lower febrile response in PVN-lesioned animals than in controls. The body temperature of PVN-lesioned animals and controls showed no difference during 300 min of exposure to heat (32 degrees C) or cold (7 degrees C). These results suggest that the PVN contributes to the complex regulation of temperature during the febrile response but not during the maintenance of normal body temperature.

Animals↗