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Biomedical subjects

A Schulz

Publications and source records attributed to A Schulz.

At least 163 records · Page 9Linked to original sources

Immunoelectron microscopy of osteonectin and type I collagen in osteoblasts and bone matrix.

The pathway of production, secretion, and extracellular deposition of type I collagen and osteonectin was studied by immunoelectron microscopy using respective polyclonal antibodies. Protein A gold and immunogold methods yielded to similar results in human callus tissue used as a model of bone formation. The intracellular distribution of osteonectin in active osteoblasts is found as a faint immunolabeling of vesicular Golgi fields and some lamellae of rough endoplasmic reticulum. A more intensive labeling occurs in opaque cytoplasmic vesicles pointing to the process of secretion as some of the vesicles are connected with the basal cell membrane. Our type I collagen antibody did not label the respective intracellular compartments. Extracellularly, the type I collagen antibody showed a continuous labeling from the immature subcellular osteoid to the mineralized bone. Osteonectin antibodies were bound first to the deeper layer of osteoid maturation with intensity increasing below the mineralization front. Osteonectin is thought to be associated with mineralization of bone matrix.

Antibodies↗

Stereospecific production of the herbicide phosphinothricin (glufosinate) by transamination: isolation and characterization of a phosphinothricin-specific transaminase from Escherichia coli.

An aminotransferase capable of transaminating 2-oxo-4-[(hydroxy)(methyl)phosphinoyl]butyric acid to L-phosphinothricin [L-homoalanine-4-yl-(methyl)phosphinic acid], the active ingredient of the herbicide Basta (Hoechst AG), was purified to apparent homogeneity from Escherichia coli K-12. The enzyme catalyzes the transamination of L-phosphinothricin and various analogs with 2-ketoglutarate as the amino group acceptor. The transaminase has a molecular mass of 43 kilodaltons by sodium dodecyl sulfate-gel analysis and an isoelectric point of 4.35. The enzyme was most active in the high-pH region, with a maximum at pH 8.0 to 9.5, and had a temperature optimum of 55 degrees C. Heat stability was observed up to 70 degrees C. Substrate specificity studies suggested that the enzyme is identical with the 4-aminobutyrate:2-ketoglutarate transaminase (EC 2.6.1.19). The first 30 amino acids of the N terminus of the protein were determined by gas phase sequencing. The transaminase was immobilized by coupling to the epoxy-activated carrier VA-Biosynth (Riedel de Haen) and used in a column reactor for the continuous production of L-phosphinothricin. The enzyme reactor was operated for 7 weeks with only a slight loss of catalytic capacity. Production rates of more than 50 g of L-phosphinothricin per liter of column per h were obtained.

Aminobutyrates↗

Stereospecific production of the herbicide phosphinothricin (glufosinate) by transamination: cloning, characterization, and overexpression of the gene encoding a phosphinothricin-specific transaminase from Escherichia coli.

We have cloned the gene encoding a 43-kilodalton transaminase from Escherichia coli K-12 with a specificity for L-phosphinothricin [L-homoalanine-4-yl-(methyl)phosphinic acid], the active ingredient of the herbicide Basta (Hoechst AG). The structural gene was isolated, together with its own promoter, and shown to be localized on a 1.6-kilobase DraI-BamHI fragment. The gene is subject to catabolite repression by glucose; however, repression could be relieved completely when 4-aminobutyrate (GABA) served as the sole nitrogen source. The regulation pattern obtained and a comparison of the restriction map of the initially cloned 15-kilobase SalI fragment with the physical map of the E. coli K-12 genome suggest that the cloned gene is identical with gabT, a locus on the gab gene cluster of E. coli K-12 which codes for the GABA:2-ketoglutartate transaminase (EC 2.6.1.19). A number of expression plasmids carrying the isolated transaminase gene were constructed. With these constructs, the transaminase expression in transformants of E. coli could be increased up to 80-fold compared with that in a wild-type control, and the transaminase constituted up to 20% of the total soluble protein of the bacteria. Thus, the protein crude extracts of the transformants could be used, after a simple heat precipitation step, for the biotechnological production of L-phosphinothricin in an enzyme reactor.

Aminobutyrates↗

Molecular analysis of two genes of the Escherichia coli gab cluster: nucleotide sequence of the glutamate:succinic semialdehyde transaminase gene (gabT) and characterization of the succinic semialdehyde dehydrogenase gene (gabD).

We have characterized two genes of the Escherichia coli K-12 gab cluster, which encodes the enzymes of the 4-aminobutyrate degradation pathway. The nucleotide sequence of gabT, coding for glutamate:succinic semialdehyde transaminase (EC 2.6.1.19), alternatively known as 4-aminobutyrate transaminase, was determined. The structural gene consists of 1,281 nucleotides specifying a protein of 426 amino acids with a molecular mass of 45.76 kDa. The protein shows significant homologies to the ornithine transaminases from Saccharomyces cerevisiae and from rat and human mitochondria. Three functionally and structurally important amino acid residues of the transaminase were identified by sequence comparison studies, and evolutionary relationships of the aminotransferases are discussed. The gabD gene, encoding succinic semialdehyde dehydrogenase (EC 1.2.1.16), was cloned and shown to be located adjacent to the 5' end of gabT. Expression studies with subfragments of the initially cloned DNA region revealed a maximal size of 1.7 kb for gabD. Both genes are cotranscribed from a promoter located upstream of gabD.

4-Aminobutyrate Transaminase↗

[Postoperative ventilation in non-intubated patients with iron lung versus conventional ventilation].

Postoperative ventilation using a steel lung is an alternative to conventional ventilation through an endotracheal cannula. Both methods were compared in two groups of patients undergoing major abdominal surgery. Group A was ventilated using a Servo Ventilator 900C, whereas group B was ventilated using a newly designed steel lung. The duration of ventilation using the steel lung and that of postoperative intubation were significantly shorter than those used in group A, in spite of a higher anesthesiological and surgical risk in patients in group B. pO2 was significantly increased during ventilation with the steel lung in comparison to conventional ventilation. An automatic classification of EEG activity was used to estimate depth of anesthesia. This showed a close correlation between clinical signs and anesthesiological levels. The spectroanalytical evaluation of ventilation curves allowed the early recognition of spontaneous breathing and determined tidal volumes using steel lung ventilation.

Aged↗

[Cyclosporine-induced gingival hyperplasia in renal transplant patients].

Cyclosporine-induced gingival hyperplasia was investigated in a clinical study of 80 renal transplant patients. 34% of the patients exhibited at least mild gingival hyperplasia in the anterior, and 9% presented this finding also in the posterior region. No direct correlation was found between the oral dose or the whole blood concentration of cyclosporine and the presence of gingival overgrowth. Young and female patients were at a significantly greater risk of developing cyclosporine-induced gingival hyperplasia than the other patients. The presence of dental plaque or gingival inflammation was not found to be related to the incidence of gingival hyperplasia.

Adolescent↗

[Ileus symptoms due to 3 distinctively differentiated small intestine carcinomas in nontropical sprue].

Recently recurring abdominal pain, bouts of diarrhoea and weight-loss of 15 kg developed in a 50-year-old woman who had for 18 years been treated by diet for nontropical sprue. She was hospitalized for signs of mechanical ileus. Radiological examination revealed three stenosing small intestine tumours, one of them obstructing the lumen. After resection of the affected segments, with end-to-end anastomosis, histological, histological examination of the surgical specimens demonstrated three small intestine carcinomas of different grades of differentiation. The largest tumour had already metastasized to four regional lymph nodes. This case illustrates the potentially precancerous nature of nontropical sprue which has been present for many years and thus requires careful follow-up.

Adenocarcinoma↗

Immunohistological demonstration of osteonectin in normal bone tissue and in bone tumors.

Osteonectin is a noncollagenous protein of bone which is believed to be bone specific, since its concentration in bone tissue is 500- to 1000-fold higher than in other connective tissues. Immunohistochemistry with polyclonal osteonectin antibodies shows a highly specific marking of actively matrix-producing osteoblasts. Osteonectin is involved in the process of osteoid maturation and mineralization. Immunohistochemistry of osteonectin antibodies proved to be a valuable tool in the diagnosis of bone tumors. Since osteonectin production seems to be an early event in the differentiation of cells of the osteoblastic lineage, immunoreactivity of osteonectin antibodies is observed in types of osteosarcoma lacking osteoid production (anaplastic, fibroblastic) while corresponding types of soft tissue tumors remain unstained.

Bone Neoplasms↗

An insulin-releasing property of imidazoline derivatives is not limited to compounds that block alpha-adrenoceptors.

As we have demonstrated previously phentolamine stimulates the release of additional insulin from isolated mouse islets and raises plasma insulin levels in the whole rat. This effect was independent of the well known property of phentolamine to block alpha-adrenoceptors. In experiments on isolated pancreatic islets from mice we now demonstrate that tolazoline and antazoline which are chemically closely related to phentolamine, share its ability to potentiate insulin release. The following results were taken as evidence that this effect does not result from an alpha-adrenoceptor blocking action of imidazoline compounds. More than 10 times higher concentrations of phentolamine were required to liberate additional insulin from isolated islets than were effective in counteracting the inhibitory effect of clonidine on insulin release. The newly introduced alpha 2-adrenoceptor antagonist BDF 8933, which is an imidazoline derivative, stimulates insulin release as well, while the irreversible alpha-adrenoceptor blocking agent benextramine of different structure failed to do so, even when being present in concentrations blocking the alpha 2-adrenoceptor-mediated effects of clonidine. Antazoline shared the ability of phentolamine to stimulate insulin release despite having no or only very little alpha-adrenoceptor blocking activity. When used under our conditions, it almost entirely failed to alleviate the inhibition of insulin release induced by clonidine. We conclude that the response of the islet cells to imidazoline derivatives is not limited to those capable of blocking alpha-adrenoceptors. On the other hand, alpha-adrenoceptor blocking agents of different chemical structure fail to induce the release of additional insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Antagonists↗

Dual action of clonidine on insulin release: suppression, but stimulation when alpha 2-adrenoceptors are blocked.

As shown previously clonidine reduces glucose-stimulated insulin release and this effect is mediated by inhibitory postsynaptic alpha 2-adrenoceptors. The present experiments demonstrate that clonidine has the additional property to also stimulate insulin release. This became evident when the alpha 2-adrenoceptors of isolated islets were blocked by benextramine, and thus protected from being stimulated by clonidine. In the presence of benextramine, clonidine no longer reduced, but on the contrary enhanced, the release of insulin in response to glucose. In control experiments benextramine by itself failed to affect insulin release from isolated islets. These results show that the imidazoline derivative clonidine shares the property of other imidazoline compounds to enhance the insulin response to glucose. All of these agents may stimulate insulin by binding to "imidazoline-preferring" sites, that clearly differ from alpha-adrenoceptors.

Adrenergic alpha-Antagonists↗

Immunocytochemical identification of osteogenic bone tumors by osteonectin antibodies.

18 bone-forming tumours and tumour-like lesions were investigated immunocytochemically for the presence of osteonectin. A group of non-bone-forming skeletal tumours (five cartilage-forming tumours, four Ewing sarcomas and five extraskeletal sarcomas) served as controls. The studies showed that osteonectin antibodies react reliably with benign and malignant bone-forming tumours (two cases of fibrous dysplasia, three osteoid osteomas, 13 osteosarcomas). This finding was supported by protein blot studies. Osteonectin is formed by cells which do not yet possess the morphological phenotype of osteoblasts and may be regarded as a "differentiation marker" of the osteoblastic lineage. Only chondroid bone (tissue in which chondrocytes were surrounded by osteoid matrix containing type I and type II collagen) showed a positive reaction. All other primary skeletal tumours and extraskeletal soft tissue tumours were completely negative.

Antibodies↗

[Ectopic gastric mucosa in the duodenal bulb].

The radiological and clinical findings of 12 patients with ectopic gastric mucosa in the duodenal bulb are presented. This is a defined disease with characteristic radiological features: multiple small nodular defects of the contrast medium of 1-3 mm diameter. Histology shows complete heterotopia. Pathogenesis and clinical significance are discussed with reference to the literature on this subject.

Adult↗

The herbicidally active experimental compound Hoe 704 is a potent inhibitor of the enzyme acetolactate reductoisomerase.

Growth inhibition of plants and bacteria by the experimental herbicide Hoe 704 (2-methylphosphinoyl-2-hydroxyacetic acid) was alleviated by the addition of the branched-chain amino acids to growth media. Hoe 704 caused a massive accumulation of acetoin and acetolactate, indicating its direct interference with the branched-chain amino acid biosynthetic pathway. The second enzyme of this pathway, acetolactate reductoisomerase (EC 1.1.1.86), was found to be subject to strong inhibition by Hoe 704. The inhibition was time-dependent and competitive with the enzyme's substrate, acetolactate. This report establishes acetolactate reductoisomerase as a new target for a herbicidal compound.

2-Acetolactate Mutase↗

Phentolamine, a deceptive tool to investigate sympathetic nervous control of insulin release.

We investigated the effects of phentolamine and another more selective alpha 2-adrenoceptor antagonist, rauwolscine, on insulin release in vivo (in female Wistar-rats) and in vitro (in perfused rat pancreas and in isolated perifused mouse islets). Phentolamine was found to significantly increase glucose-induced insulin release. On the other hand, rauwolscine failed to do so, when applied in a concentration that effectively antagonized the inhibitory effect of clonidine. These results demonstrate that phentolamine is capable of directly stimulating insulin release. This effect is thus not mediated by alpha-adrenoceptors. For this reason phentolamine is not an appropriate tool to study possible inhibitory effects of the sympathetic nervous system on insulin release. An enhanced insulin response as may be observed in animals and in man in the presence of phentolamine does not furnish evidence for a tonic inhibitory control of the islet cells by the sympathetic nervous system.

Animals↗

Microcalorimetric investigations on Listeria.

Sixty-one Listeria strains--Listeria monocytogenes (35 strains), Listeria ivanovii (8), Listeria innocua (7), Listeria welshimeri (6) and Listeria seeligeri (5)--were tested microcalorimetrically for their heat production upon growth in Columbia broth. Listeria ivanovii and Listeria seeligeri displayed different and quite characteristic thermograms. Listeria welshimeri, Listeria innocua and all but one of the Listeria monocytogenes strains showed similar microcalorimetric curves, which differed considerably from those of Listeria ivanovii and Listeria seeligeri.

Calorimetry↗