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M Schindler

Publications and source records attributed to M Schindler.

At least 55 records · Page 3Linked to original sources
PubMed1999

Immunohistochemical localization of the somatostatin sst2(b) receptor splice variant in the rat central nervous system.

Somatostatin is a neuromodulator in the mammalian CNS. To date, genes for at least five different somatotrophin release inhibiting factor receptors, termed sst1-sst5, have been cloned. The rat sst2 receptor exists in two splice variants, sst(alpha)a) and sst2(b), which differ in their carboxy-termini. When heterologously expressed in Chinese hamster ovary-K1 cells, these splice variants show little difference in their operational characteristics. Recently, the distribution of the sst2(a) receptor was documented, yet at present no data are available about the distribution of the sst2(b) receptor in the CNS. Here, we present the characterization of a novel polyclonal anti-peptide antibody that is selective for the sst2(b) receptor splice variant. The antibody was raised against the unique intracellular carboxy-terminal portion of the receptor protein. Using this affinity-purified antibody in western blotting experiments, the sst2(b) receptor expressed in Chinese hamster ovary-K1 cells was shown to be a glycoprotein with a molecular weight centred at about 85,000. The antibody showed no cross-reactivity to any of the recombinant human sst1-5 receptors, the rat sst2(a) receptor or wild-type Chinese hamster ovary-K1 cells. Employing immunohistochemistry, we investigated the distribution of the sst2(b) receptor in the brain and spinal cord of adult rats. A distinct distribution was found throughout the rostrocaudal axis of the CNS. Somatodendritic as well as axonal staining was observed. Somatodendritic labelling was particularly obvious in the olfactory bulb, cerebral cortex, hippocampal formation, mesencephalic trigeminal nucleus and cerebellum, as well as in cranial and spinal motor areas. The results show that the distribution of the sst2(b) receptor partially overlaps with that of the sst2(b) receptor, although there were differences in a number of brain areas. The location of the sst2(b) receptor implies that it may mediate a modulatory role of somatostatin inhibitory releasing factor on sensory as well as motor functions.

Animals↗
PubMed1999

Immunohistochemical expression of somatostatin type 2A receptor in neuroendocrine tumors.

Somatostatin (SS) and SS analogues inhibit the growth of various kinds of endocrine and exocrine cells via the SS receptor (SSTR). Carcinoid tumor is representative of the tumors treatable by SS analogues. We examined the expression of SSTR2A by immunohistochemical and in situ hybridization methods with a specific antibody against a synthesized 20-amino acid peptide of the COOH terminus of human SSTR2A and oligonucleotide probes in 62 endocrine tumors of various kinds: pancreatic endocrine tumor; carcinoid; neuroendocrine carcinoma; medullary thyroid carcinoma; pheochromocytoma; and small cell carcinoma of the lung, neuroblastoma, and ganglioneuroma. SSTR2A was expressed in 87% of these tumors and at both primary and metastatic sites. The immunohistochemical reactivity of SSTR2A was strong on the cell membrane and less intense in the cytoplasm of the tumor cells. SSTR2A mRNA was also detected in the tumor cells. The results indicate the usefulness of SSTR2A analogues for the treatment of neuroendocrine tumors, even metastatic ones: metastatic carcinoids, metastatic pheochromocytomas, tumors that adhered to large vessels, and neuroendocrine carcinomas.

Adrenal Gland Neoplasms↗
PubMed1998

Identification of somatostatin sst2(a) receptor expressing neurones in central regions involved in nociception.

Somatostatin is a neuromodulator and neurotransmitter in the central nervous system. Administration of somatostatin to the spinal cord or brain areas involved in nociception has been shown to result in analgesia. Little information is available about the somatostatin receptor types which may be involved in mediating the neuromodulatory and analgesic effects of the peptide. To define the neuronal systems expressing the sst2(a) receptor in brain areas associated with analgesia, immunohistochemical co-localisation studies were carried out in the periaqueductal grey (PAG) and spinal cord using an antibody specific for the sst2(a) receptor. To further define sst2(a) receptor expressing neurones, sst2(a) receptor immunohistochemistry was combined with retrograde tracing using fluorogold. In the PAG, sst2(a) receptor expressing neurones were found to co-express calbindin D28k (36%), the glutamate transporter EAAC-1 (25%), and GABA transporter GAT-1 ( approximately 10%). A total of 65% of sst2(a) positive neurones projected to the thalamus. In the spinal cord, the sst2(a) receptor shows cellular co-localisation with EAAC-1 and GAT-1. Immunohistochemistry and receptor autoradiography using [125I]BIM 23027 after dorsal rhizotomy of the lumbar dorsal roots, L4 and L5, suggests that the somatostatin sst2(a) receptor is not present on primary afferent neurones. Dorsal hemisections of the mid thoracic cord did not alter the immunohistochemical signal for the somatostatin sst2(a) receptor, providing further evidence for an intrinsic localisation of the receptor protein in the dorsal horn of the spinal cord. These data show that the somatostatin sst2(a) receptor exists on morphologically and neurochemically heterogenous neurones and is closely associated with brain areas involved in analgesia and the modulation of nociception.

Animals↗
PubMed1998

Defective acidification in human breast tumor cells and implications for chemotherapy.

Multidrug resistance (MDR) is a significant problem in the treatment of cancer. Chemotherapeutic drugs distribute through the cyto- and nucleoplasm of drug-sensitive cells but are excluded from the nucleus in drug-resistant cells, concentrating in cytoplasmic organelles. Weak base chemotherapeutic drugs (e.g., anthracyclines and vinca alkaloids) should concentrate in acidic organelles. This report presents a quantification of the pH for identified compartments of the MCF-7 human breast tumor cell line and demonstrates that (a) the chemotherapeutic Adriamycin concentrates in acidified organelles of drug-resistant but not drug-sensitive cells; (b) the lysosomes and recycling endosomes are not acidified in drug-sensitive cells; (c) the cytosol of drug-sensitive cells is 0.4 pH units more acidic than the cytosol of resistant cells; and (d) disrupting the acidification of the organelles of resistant cells with monensin, bafilomycin A1, or concanamycin A is sufficient to change the Adriamycin distribution to that found in drug-sensitive cells, rendering the cell vulnerable once again to chemotherapy. These results suggest that acidification of organelles is causally related to drug resistance and is consistent with the hypothesis that sequestration of drugs in acidic organelles and subsequent extrusion from the cell through the secretory pathways contribute to chemotherapeutic resistance.

Anti-Bacterial Agents↗
PubMed1998

Localization of the somatostatin sst2(a) receptor in human cerebral cortex, hippocampus and cerebellum.

The distribution and cellular localization of the somatostatin sst2(a) receptor was investigated in selected human brain areas using an anti-peptide antibody raised against a carboxy-terminal portion of the receptor protein. The sst2(a) receptor was found to be present on neurones and processes in the deep layers of the cerebral cortex, in the subicular complex and the hippocampal formation. Further signals were obtained in the molecular and granular layer of the cerebellum, with occasional weakly stained Purkinje cells. The regional distribution of the receptor protein was compared with quantitative autoradiography using a sst2 receptor selective ligand [125I]BIM23027.

Autoradiography↗
PubMed1998

5-HT1B receptor-mediated contractions in human temporal artery: evidence from selective antagonists and 5-HT receptor mRNA expression.

1. In the human temporal artery both 5-HT1-like and 5-HT2A receptors mediate the contractile effects of 5-hydroxytryptamine (5-HT) and we have suggested that the 5-HT1-like receptors resemble more closely recombinant 5-HT1B than 5-HT1D receptors. To investigate further which subtype is involved, we investigated the blockade of 5-HT-induced contractions by the 5-HT1B-selective antagonist SB-224289 (2,3,6,7-tetrahydro-1'-methyl-5-[2-methyl-4'[(5-methyl-1,2,4-oxadiazole- 3-yl) biphenyl-4-yl] carbonyl] furo[2,3-f]indole-3-spiro-4'-piperidine oxalate) and the 5-HT1D-selective antagonist BRL-15572 (1-phenyl-3[4-3-chlorophenyl piperazin-1-yl] phenylpropan-2-ol). We also used RT-PCR to search for the mRNA of 5-HT1B, 5-HT1D and other 5-HT receptors. 2. The contractile effects of 5-HT in temporal artery rings were partially antagonized by SB-224289 (20, 200 nM) (apparent KB = 1 nM) and ketanserin (1 microM) but not by BRL-15572 (500 nM). 3. Sumatriptan evoked contractions (EC50, 170 nM) that were resistant to blockade by BRL-15572 (500 nM) but antagonized by SB-224289 (20, 200 nM). 4. The potency of 5-HT (EC50) was estimated to be 94 nM for the ketanserin-sensitive receptor and 34 nM for the SB-224289-sensitive receptor. The fraction of maximal 5-HT response mediated through SB-224289-sensitive receptors was 0.20-0.67, the remainder being mediated through ketanserin-sensitive receptors. 5. We detected arterial receptor mRNA for the following receptors (incidence): 5-HT1B (8/8), 5-HT1D (2/8), 5-HT1F (0/4), 5-HT2A (0/8) 5-HT2B (0/8), 5-HT2C (0/8), 5-HT4 (4/8) and 5-HT7 (4/8). 6. We conclude that the ketanserin-resistant fraction of the 5-HT effects and the effects of sumatriptan are mediated by 5-HT1B receptors. The lack of antagonism by BRL-15572 rules out 5-HT1D receptors as mediators of the contractile effects of 5-HT and sumatriptan.

Adolescent↗
PubMed1998

Molecular cloning and functional characterization of a rat somatostatin sst2(b) receptor splice variant.

1. The mouse somatostatin (SRIF) sst2 receptor exists in two splice variants, sst2(a) and sst2(b), which differ in their intracellular carboxy-termini only. The murine sst2(b) receptor was reported to be less prone to agonist-induced desensitization as compared with the sst2(a) receptor. To determine whether a sst2(b) splice variant with similar functional characteristics exists in the rat, we have isolated a cDNA fragment from rat gastric mucosa encoding a sst2(b) receptor and expressed the full-length protein in CHO-K1 cells for functional characterization. 2. This study provides the first evidence for the occurrence in the rat of the sst2(b) receptor, which has a 15 amino acid carboxy-terminus differing in composition to the 38 amino acid C-terminus of the rat sst2(a) receptor. 3. In CHO-K1 cells expressing rat recombinant sst2(a) or sst2(b) receptors, SRIF caused concentration-dependent increases in extracellular acidification rates (EAR) with pEC50 values of 9.0 and 9.9, respectively. Pre-treatment with pertussis toxin (Ptx) caused a rightward displacement of the SRIF concentration-effect curves with pEC50 values of 8.3 (sst2(a) and 8.4 (sst2(b)). 4. SRIF (3 pM-3 nM) also caused concentration-dependent inhibition of forskolin-stimulated cyclic AMP formation in CHO-sst2(a) cells (pIC50 10.5) and CHO-sst2(b) cells (pIC50 10.4). The degree of inhibition was less with higher concentrations of SRIF resulting in bell-shaped concentration-effect curves. Following pre-treatment with Ptx, the inhibitory effect of SRIF was abolished and SRIF caused only increases in cyclic AMP formation. 5. Both the SRIF-induced increases in EAR and inhibition of cyclic AMP formation were susceptible to agonist-induced desensitization, but this was less apparent following pre-treatment with Ptx. 6. This demonstrates that the operational characteristics of the recombinant rat sst2(a) and sst2(b) receptors are broadly similar. Both isoforms couple to Ptx-sensitive as well as -insensitive G proteins and are equally prone to agonist-induced desensitization.

Adenylyl Cyclases↗
PubMed1998

Rat somatostatin sst2(a) and sst2(b) receptor isoforms mediate opposite effects on cell proliferation.

We have investigated the actions of somatostatin (SRIF) and angiopeptin on cell proliferation of CHO-K1 cells expressing the recently cloned rat sst2(b) receptor (CHOsst2(b)) and compared these to their effects in cells expressing the sst2(a) receptor (CHOsst2(a)). In contrast to the sst2(a) receptor, the sst2(b) receptor did not mediate inhibition of bFGF (10 ng ml(-1))-stimulated re-growth and cell proliferation. Rather, SRIF (0.1-1000 nM) and angiopeptin (0.1-1000 nM) stimulated basal re-growth and proliferation of CHOsst2(b) cells in a concentration-dependent manner (estimated pEC50 values of 7.8 and 7.9, respectively). The opposite effects of SRIF on cell proliferation mediated through the two sst2 receptor isoforms were both abolished by 18 h pre-treatment with pertussis toxin. The proliferative effect via the sst2(b) receptor was also abolished by the tyrosine kinase inhibitor, genistein. In conclusion, the present study shows that the rat sst2(a) and sst2(b) receptor splice variants mediate opposite effects on cell proliferation.

Animals↗
PubMed1998

The lateralizing value of ictal clinical symptoms in uniregional temporal lobe epilepsy.

In order to assess the lateralizing value of several ictal and postictal clinical symptoms in temporal lobe epilepsy (TLE), we analyzed 89 seizures of 20 left dominant patients with intractable left (n = 9) versus right (n = 11) TLE who had undergone successful anterior temporal lobectomy. In left TLE, movement arrest at seizure onset, postictal dysphasia > 120 s and postictal dyslexia > 180 s were the most typical findings and associated with a sensitivity of 94, 94, and 100%, respectively. The highest specificity of 100% each was evident for contralateral versions of eyes and head and dystonic posturing. In right TLE, the highest sensitivity was seen for whole-body movements at seizure onset, postictal dysphasia < 120 s and postictal dyslexia < 180 s with figures of 82, 87, and 93%, respectively. As compared to left TLE, contralateral version and dystonic posturing, ictal speech, and postictal dyslexia < 180 s each had a specificity of 100%. The careful combined analysis of certain ictal clinical signs combined with consistent findings of interictal EEG and neuroimaging studies may be often sufficient to proceed with epilepsy surgery without invasive recordings even if ictal scalp EEG is not unambiguous.

Adolescent↗
PubMed1997

[Successful treatment of fulminant myocarditis with the biventricular MEDOS Assist System (MEDOS HIA-VAD)].

BACKGROUND: Successful weaning from biventricular mechanical support with full recovery of the myocardial function is extremely rare in fulminant myocarditis. We report on our experience with the new MEDOS HIA ventricular assist device. METHODS AND RESULTS: We used the MEDOS assist system to support a 30-year-old woman with profound circulatory impairment caused by acute myocarditis. The device provided adequate hemodynamics and recovery of myocardial function. Despite anticoagulation therapy we had to change either the left or right ventricular pump chamber because of clot formation on the surface of the outflow tract. On the 14th postoperative day a surgical reintervention was necessary for bleeding from the cannulation site of the pulmonary artery. After 17 days the myocardial function had recovered and we could remove the assist system. The following parameters were measured before implantation of the MEDOS assist system and after weaning from circulatory support: ejection fraction 15 vs. 45%, cardiac index 0.7 vs. 2.6 L/min/m2, arterial pressure (systolic/diastolic/mean) 81/55/66 vs. 113/66/82 mm Hg, pulmonary artery pressure 33/25/29 vs. 34/20/28 mm Hg, pulmonary capillary wedge pressure 24 vs. 19 mm Hg. CONCLUSIONS: Despite severe cardiac failure in fulminant myocarditis requiring biventricular mechanical support full recovery of the myocardium is possible.

Adult↗
PubMed1997

MEDOS HIA-VAD biventricular assist device for bridge to recovery in fulminant myocarditis.

Successful weaning from biventricular mechanical support with full recovery of the myocardial function is extremely rare in fulminant myocarditis. We report on our experience with the MEDOS HIA-VAD ventricular assist device. The device worked for 17 days and provided adequate hemodynamics. Despite anticoagulation therapy, we had to change both ventricles because of clot formation on the surface of the outflow tract. After 17 days the myocardial function had recovered and we could remove the assist system.

Acute Disease↗
PubMed1997

Immunohistochemical localization of the somatostatin SST2(A) receptor in the rat brain and spinal cord.

The neuropeptide somatostatin is widely distributed in the CNS and is believed to play a role as a neurotransmitter or a neuromodulator. Somatostatin mediates its actions by the binding of the peptide to high affinity membrane receptors. The genes for five somatostatin receptor types have been cloned recently and Northern blotting and in situ hybridization studies have shown that the transcripts of all five types are expressed in the CNS. Here we report the cellular distribution of somatostatin sst2(a) receptor protein in the adult rat CNS, using a polyclonal anti-peptide antibody directed against a portion of the C-terminal domain of the receptor. The specificity of the affinity-purified antibody was demonstrated by Western blotting and immunolabelling of cells transfected with a hemagglutinin epitope-tagged version of the sst2(a) receptor. Immunohistochemistry showed a distinct distribution of the receptor protein in the rat brain. Cells and processes were labelled in a number of areas, including the basolateral amygdala, the locus coeruleus, the endopiriform nucleus, the deep layers of the cerebral cortex, the subiculum, the claustrum, the habenula, the interpenduncular nucleus, the hippocampus and the central grey. In the spinal cord, the substantia gelatinosa showed strongly-labelled cell bodies and their processes. This study provides an improved understanding of the distribution of the sst2(a) receptor in rat brain.

Animals↗
PubMed1997

Penetration of teicoplanin into heart valves and subcutaneous and muscle tissues of patients undergoing open-heart surgery.

Penetration of teicoplanin into serum, heart valves, and subcutaneous and muscle tissues was determined in 22 patients undergoing open-heart surgery. Each patient received 12 mg of teicoplanin per kg of body weight as a 30-min intravenous infusion preoperatively. Within 10 h, serum concentrations of teicoplanin declined from 43.1 to 2.8 microg/ml. Teicoplanin concentrations in subcutaneous tissues reached their peak of 9.2 microg/g after 2 to 3 h and decreased slowly to 2.3 microg/g after 9 to 10 h. Concentrations in muscle decreased from 8.7 microg/g to nondetectable levels. Teicoplanin concentrations in cardiac valvular tissue reached their peak of 6.1 microg/g and decreased thereafter to 1.7 microg/g. Teicoplanin concentrations in heart valves were high enough to inhibit methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci, which are known to cause postoperative wound infections and infective endocarditis.

Aged↗
PubMed1996

Different inducibility of expression of the two xylanase genes xyn1 and xyn2 in Trichoderma reesei.

Regulation of formation of the extracellular xylanase system of Trichoderma reesei QM 9414 during growth on xylan, cellulose, and replacement onto a number of soluble inducers was investigated by Northern analysis of xyn1 and xyn2 transcripts and by the use of the Escherichia coli hph (hygromycin B-phosphotransferase-encoding) gene as a reporter. Whereas the xyn1 promoter is active in the presence of xylan and xylose, and virtually silenced in the presence of glucose, the xyn2 promoter enables basal transcription at a low level, but is enhanced in the presence of xylan and xylobiose and also of sophorose or cellobiose. The respective regulatory nucleotide regions were localized on a 221-base pair fragment and a 55-base pair fragment of the xyn1 and xyn2 5'-upstream noncoding sequences, respectively. Electrophoretic mobility shift assays, using cell-free extracts, identified induction-specific protein-DNA complexes: one complex of high mobility was observed under basal, noninduced conditions (glucose) with xyn2, which was in part replaced by a slow-migrating complex upon induction by xylan or sophorose. Both complexes bound to a CCAAT box. With xyn1, the induced complex also binds to a CCAAT box, but this binding is not observed in the presence of the carbon catabolite repressor Cre1, which binds to a nearby located consensus motif.

Base Sequence↗