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U Sommer

Publications and source records attributed to U Sommer.

At least 37 records · Page 2Linked to original sources

Establishing an algorithm for molecular genetic diagnostics in 127 families with juvenile nephronophthisis.

BACKGROUND: Juvenile nephronophthisis (NPH1), an autosomal recessive cystic disease of the kidney, represents the most common genetic cause of end-stage renal disease in the first two decades of life. On the basis of identification of the gene (NPHP1) defective in NPH1 and the presence of homozygous deletions of NPHP1 in the majority of NPH1 patients, molecular genetic diagnosis for NPH1 is now possible. Molecular genetic testing offers the only method for definite diagnosis of NPH1 and avoids invasive diagnostic measures like renal biopsy. METHODS: We examined 127 families (204 patients) with the presumed diagnosis of NPH using molecular genetic diagnostic techniques. In 68 families, renal biopsy was performed and was consistent with NPH, and in 61 families, there was more than one affected child ("multiplex families"). RESULTS: In 74 families (115 patients), there was proof of the diagnosis of NPH1 by detection of a homozygous deletion of the NPHP1 gene, and in 5 families a heterozygous deletion in combination with a point mutation in NPHP1 was demonstrated. Furthermore, for 16 families, NPH1 was excluded with high likelihood by linkage analysis, and for 20 families by detection of heterozygosity for two newly identified polymorphic markers within the deletion region. In 5 of the remaining 12 families, which were noninformative for these markers, fluorescence in situ hybridization did not detect any further heterozygous deletions. CONCLUSIONS: The diagnosis of NPH1 was proven by molecular genetic techniques in 62% of families with one or more children with the presumed diagnosis of NPH. We present evidence that there is a fourth locus for NPH, since only 6 of the 26 multiplex families in whom the diagnosis of NPH1 was excluded were compatible with linkage to other loci for NPH. On the basis of the presented data, we propose an algorithm for molecular genetic diagnostics in NPH.

Adaptor Proteins, Signal Transducing↗

Proliferation in the gastric epithelium of bovine abomasum during foetal development as revealed by Ki-67 immunocytochemistry.

For the first time, proliferative cells in the abomasal mucosa of 30 bovine foetuses with a crown-rump length of 34 to 1000 mm (approximately 45-250 days of gestation) were detected using the antibody MIB-1 against the (human) nuclear-associated protein Ki-67. At the beginning of mucosal development, the epithelium is stratified and MIB-1 positive cells are scattered all over the epithelium. With the formation of gastric pit/gland invaginations, the proliferative cells are mostly confined to the basal half or third of the gastric pit/gland unit, both in the regio glandulae propriae and in the regio glandulae pyloricae. From the 27th week in the regio glandulae propriae and the 30th week in the pyloric gland region, respectively, the proliferative cells move upwards and are situated in the bottom of the pit and the upper part of the gland, whereas the base of the gland is free of labelling.

Abomasum↗

Termination of IL-6-induced STAT activation is independent of receptor internalization but requires de novo protein synthesis.

The interleukin-6 (IL-6) receptor complex comprises the IL-6 receptor (IL-6R, gp80) and the signal transducer gp130. Binding of IL-6 to its receptor results in dimerization of gp130, activation of the Jak/STAT pathway, and in a down-regulation of IL-6 binding sites by endocytosis. The STAT activation after stimulation is transient, being maximal after 15-30 min and disappearing after 60-90 min. The mechanism which leads to the termination of the signal is still unknown. In this paper we have studied whether the down-modulation of the STAT signal requires the endocytosis of the receptor complex. Our results suggest that the desensitization of the IL-6 signal is not due to internalization of the receptor complex but requires de novo protein synthesis.

Animals↗

Workshop IV: drug treatment guidelines for the long-term management of Parkinson's disease.

An attempt was made to establish a decision algorithm for the treatment of idiopathic Parkinson's disease at various stages and in different subgroups such as akinetic-rigid or tremor dominance type. We suggest treating young patients with selegiline and a dopamine agonist. In the tremor dominance type we use either budipine or a dopamine agonist. Due to levodopa-induced dyskinesia, we try to avoid levodopa in the early stages of the disease and use it only later in more advanced situations in a combination therapy with dopamine agonists. Since IPS is not only based upon dopamine deficiency but also on resulting glutamatergic overstimulation, we advocate the use of a glutamate antagonist such as amantadine or budipine. Catechol-O-methyl inhibitors are very helpful when wearing-off occurs. Anticholinergics are only used in the early stages of tremor-dominant IPS because we fear enhancing the risk of dementia.

Adult↗

The sialate pyruvate-lyase from pig kidney: purification, properties and genetic relationship.

For further insight into the structural relationship between mammalian and microbial sialate pyruvate-lyases, the enzyme from pig kidney was purified to homogeneity from the tissue homogenate by a heat precipitation step followed by anion exchange and Hydrophobic Interaction Chromatography or native gel electrophoresis, respectively. The pure enzyme preparation exhibited an about 1000-fold increase of specific activity compared to the supernatant after the first centrifugation and revealed a single band at 34-37 kDa after SDS-PAGE, which represents the monomeric form of the protein. While the native enzyme seems to be a trimer according to the molecular weight obtained by gel filtration (108 kDa), crosslinking with dimethylpimelimidate suggests it to be a tetramer. The lyase is optimally active at about 75 degrees C and in the pH range of 7.6 to 8.0 and belongs to the class I-aldolases, due to its non-requirement of metal ions and the presence of lysine as the main functional residue in its catalytic centre. These data are similar to those obtained with bacterial lyases. However, peptide fragments of this enzyme show less similarity to primary lyase structures of microbia than to those derived from expressed sequence tags of mammals.

Amino Acid Sequence↗

The terminal enzymes of sialic acid metabolism: acylneuraminate pyruvate-lyases.

The acylneuraminate pyruvate-lyase gene from Clostridium perfringens was sequenced and found to be most similar to the lyase gene from Haemophilus influenzae. Both the recombinant clostridial enzyme and the native enzyme from pig kidney were purified in larger amounts and characterized. The properties of the porcine lyase are similar to the microbial ones. However, the much higher degree of similarity in comparison to the microbial enzymes that was found between porcine lyase peptides and two putative mammalian lyase sequences show that the latter form an own group apart from the microbial lyases. Actual models of the acylneuraminate pyruvate-lyase reaction are discussed.

Amino Acid Sequence↗

[Structure of infrapatellar synovial ligament in dogs].

The knee joint of the dog is frequently the subject of surgery if the cruciate ligaments or the menisci are injured. In the knee joint of the dog we find a ligament, which is spread out between the anterior part of the fossa intercondylaris ossis femoris and the corpus adiposus infrapatellaris. This ligamentum synoviale infrapatellare has not previously been described and represents a homologous structure to the plica synovialis infrapatellaris of human knee joint. Just like the latter and just like the cruciate ligaments it is a rudiment of a septum articulare genu, which is spread out between an articulatio femorotibialis and an articulatio femorofibularis in the early phylogenesis of tetrapods. The ligamentum synoviale infrapatellare of the dog consists of a stratum synoviale and a stratum spinosum, which includes blood vessels, nerves, many collagenous fibres and some elastic fibres. We do not find evidence for sensory nerve endings or corpuscular receptors. Therefore a servering of the ligament during knee joint surgery does not affect the proprioceptive potential of the knee joint.

Animals↗

Glucocorticoids regulate the expression of the human osteoblastic endothelin A receptor gene.

The endothelial cell-derived peptide endothelin 1 (ET1) stimulates cell proliferation and differentiated functions of human osteoblastic cells (HOC), and HOC constitutively express the endothelin A receptor (ETRA). Therefore, ET1 may play an important role in the regulation of bone cell metabolism. As glucocorticoids (GC) exert a profound influence on bone metabolism and increase the effects of ET1 on bone cell metabolism in vitro, the effects of GC on ETRA expression in HOC were investigated. Dexamethasone (DEX) increased ETRA mRNA levels in a dose- and time-dependent fashion. The effects of dexamethasone, prednisolone, and deflazacort on the increase of ETRA mRNA levels correlate positively with their binding affinity to the GC receptor. Scatchard analysis of ET1 binding data to HOC revealed that DEX increased the binding capacity for ET1 from 25,300 to 62,800 binding sites per osteoblastic cell, leading to an enhanced mitogenic effect of ET1 on HOC after preincubation with DEX. Transiently transfected primary HOC with a reporter gene construct, containing the 5'-flanking region of the ETRA gene fused to luciferase gene, showed a promoter-dependent expression of the reporter gene and the induction of reporter gene expression by DEX treatment. Total RNA extracts of femoral head biopsies with osteonecrotic lesions from GC-treated patients showed threefold higher ETRA mRNA levels compared with extracts of bone biopsies from patients with traumatically induced osteonecrosis and coxarthrosis. Furthermore, GC treatment increased plasma ET1 levels by 50% compared with pretreatment values. These findings suggest that GC induced upregulation of ETRA, and ET1 plasma levels enhance ET1's anabolic action on bone cell metabolism. Increased ET1 concentrations may also impair bone perfusion by vasoconstriction in a metabolically activated skeletal region.

Adult↗

Regulation of interleukin-6 expression in human osteoblastic cells in vitro.

Interleukin-6 (IL-6) is a potent stimulator of bone resorption which has been demonstrated in a variety of in vivo and in vitro models. We investigated the regulation of IL-6 secretion in primary human osteoblastic cells (HOC) in vitro by cytokines known to play an important role in coupling bone formation to bone resorption. HOC were isolated from healthy adults who underwent selective orthopedic surgery and treated with cytokines released in the bone microenvironment during coupling i.e Interleukin-1beta (IL-1beta), Tumor Necrosis Factor alpha (TNFalpha), Transforming Growth Factor beta1 and 2 (TGFbeta 1 and 2) and Endothelin-1 (ET-1). Furthermore, we determined whether systemically-acting steroid hormones of gonadal and adrenal origin as well as glucocorticoids affect the local regulation of IL-6 secretion in primary HOC. To examine the effects of different steroid hormones on IL-6 production, HOC were exposed to estradiol (E2), dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA) and dexamethasone (Dexa) with and without a subsequent treatment of the HOC populations with cytokines. We observed that (1) IL-1beta and TNFalpha induced IL-6 in a dose and time-dependent fashion, (2) TGFbeta 1 and 2 enhanced basal and IL-1beta and TNFalpha induced IL-6 expression, (3) ET-1 elicited a dose-dependent stimulatory effect on IL-6 expression. (4) E2, DHT and DHEA alone and in combination with IL-1beta and TNFalpha elicited no reproducible dose-dependent effect on IL-6 production, whereas Dexa inhibited basal and IL-1beta and TNFalpha induced IL-6 expression dose dependently. In conclusion, IL-1beta, TNFalpha, TGFbeta 1 and 2 and ET-1 may participate in the regulation of bone resorption by stimulating IL-6 expression in HOC. Dexa inhibits the constitutive and cytokine stimulated IL-6 expression, whereas there is no in vitro evidence that sex steroids exert a major inhibitory effect on the osteoblastic secretion of IL-6 as demonstrated in a primary human bone cell model.

Aged↗

Electron-microscopic detection of cytoskeletal actin in the epithelial cells of bovine abomasum.

The fine structural distribution of cytoskeletal actin was investigated in the oxyntic gland region and in the pyloric gland region of bovine abomasal mucosa using immunoelectron methods. In all exocrine cell types of the gastric epithelium, actin filaments exhibit a polarized pattern of distribution. In the members of the mucus- and protein-synthesizing cell line, the actin filaments form a prominent barrier to the exocytosis of secretory granules underneath the apical membrane. Therefore, actin is supposed to be involved in the control of exocytosis. The basolateral subplasmalemmal cortex of the exocrine cells, however, shows weaker labeling for actin filaments. In this position actin might be responsible for the movement of epithelial cells along the length of the gastric gland during physiological cell renewal and for the maintenance of the integrity of the epithelial unit and of cell polarization. In parietal cells of bovine abomasum, actin filaments are, in addition, associated with the intracellular canaliculus, but not with the tubulovesicular compartment. We assume that actin filaments reorganize the canalicular membrane after membrane translocations in the course of acid secretion. For the first time, actin filaments were documented in the rare brush cell type of bovine abomasum, where they do not form a terminal web beneath the apical membrane and therefore point out the nonexocrine function of the brush cell type. In all polarized cells of the bovine abomasum, actin filaments underlay the two apical members of the junctional complex, the tight junction and the adherens junction, where they might be involved in the regulation of cell adhesion, cell motility and cell shape determination.

Abomasum↗

[Intraepithelial nerve endings in the lingual marginal papillae of newborn piglets].

The structure and occurrence of nerve fibres and nerve terminals in the lingual marginal papillae of newborn piglets are investigated by means of the osmium-zinc-iodate method and by transmission-electron microscopy. Subepithelial nerve fibres primarily accompany the blood vessels. They cross the epithelium with an almost vertical course to terminate in the stratum superficiale or the stratum corneum. Due to rows of varicosities, the axons look like a 'string of pearls'. As a result of the superficial localization and the typical ultrastructure, they seem to have a sensory function. Therefore, the papillae marginales are considered to function as mechanoreceptors.

Animals↗

Cytoarchitecture of the bovine pyloric epithelium during early ontogenesis.

The cytoarchitecture of the pyloric gland region in the early ontogeny of the bovine abomasal mucosa was investigated using light and transmission electron microscopic methods. Two cell types are involved in forming the lamina epithelialis in the youngest fetus with a 24 mm crown-rump-length (CRL): the indifferent cell and the endocrine cell. With a CRL of 71 mm, two other cell types occur, the granule-containing cell and the brush cell. The indifferent cell represents the first stem cell, which develops into the secretory granule-containing cell type. This seems to be the secondary stem cell lining the top of the epithelium as well as the base of the primitive epithelial tubes, and it differentiates into the surface mucous cell and the pylorocyte. Endocrine cells appear as open and closed types and represent the most differentiated cells already present in the youngest specimen in this investigation. The most rare cell type, the brush cell, appears in the bovine abomasal ontogeny much earlier than in other mammalian species.

Animals↗

[The spermathecal epithelium of the queen bee (Apis mellifera): morphology, age-dependent changes and cell contacts].

The spermatheca of the honey bee queen is covered by a single-layered, uniform, polarised epithelium. The apical cell surface is greatly enlarged by protrusions and plasma membrane infoldings, the basal cell surface by numerous interdigitating, long, small processes. Cytoplasmic organelles are chiefly represented by mitochondria. Numerous microtubuli extend throughout the cytoplasm. Golgi and endoplasmic profiles are rare. The cells are subject to senile degeneration: with increasing age, a variety of cytoplasmic inclusions appear, among which are myelinated membranes, dense bodies and dense filamentous aggregates. The spermathecal epithelium does not seem to be involved in exocrine secretion related to nutrition of the long-term stored spermatozoa. The ultra-structure points, however, to ion transport functions and to an engagement in the maintenance of an adequate physicochemical environment ensuring the viability of the spermatozoa. Cellular junctions are represented by luminal zonulae adherentes, focal cell-cell adhering junctions and hemiadhering junctions along the basal plasmalemma. Desmosomal contacts and cytoskeletal intermediate filaments are missing. Along the lateral plasmalemma, gap junctions and septate junctions are found.

Animals↗

[The differentiation of the surface mucous-cell line in the abomasum of the adult cow].

The differentiation of the surface mucous-cell lineage during physiological cell renewal was investigated using light and electron microscopy in the abomasal mucosa of adult cattle. The surface mucous cells constitute a morphologically and functionally heterogeneous population, whose members correspond to different developmental stages, OFFanged in a distoproximal gradient from the depth of the pit towards the free luminal surface. The cell lineage comprises immature pre-pit cells near the proliferative isthmus, mature pit cells within the foveola, and older interfoveolar cells lining the free surface. Ultrastructurally, differentiation can be traced towards a predominantly mucus-producing cell type and finally towards a surface-protective cell variant, which degenerates in situ and is extruded into the lumen without affecting epithelial integrity.

Abomasum↗

Differential effects of glucocorticoids on human osteoblastic cell metabolism in vitro.

Clinical observations suggest that the onset and severity of glucocorticoid (GC) induced osteoporosis is dependent on the duration of the GC treatment and the applied GC compound. To test whether these in vivo observations are reflected by different in vitro effects of various synthetic GCs on human bone cell metabolism we isolated human osteoblast-like cells (HOC) from bone biopsies of healthy (no clinical symptoms of arthritis or arthrosis) adults who underwent selective orthopedic surgery. HOC were identified as bone cells by 1,25-vitamin D3-stimulated increase of specific alkaline phosphatase (ALP) activity, secretion of osteocalcin and type-I procollagen peptide, and the ability to form mineral in vitro. We investigated the effects of dexamethasone (dexa), methylprednisolone (mpred), prednisolone (pred), and deflazacort (defla) on DNA-synthesis, ALP, and osteocalcin (OC)- and type-I procollagen peptide secretion of HOC in vitro. In summary, (1) GC exposure stimulates DNA synthesis after 6-12 hour treatment periods; (2) dex and mpred strongly inhibit DNA (48-hour treatment) and collagen synthesis but stimulate ALP, whereas pred and defla exhibit smaller effects on DNA synthesis, ALP, and collagen production; and (3) all tested glucocorticoids inhibit OC secretion by HOC in vitro. Thus, the effect of GC on DNA synthesis of HOC varies with the duration of GC exposure, and dex and mpred more potently affect HOC metabolism in vitro than pred and defla.

Adult↗

Differentiation of oxyntic cells during early ontogenesis in the bovine.

The origin and the differentiation of oxyntic cells in fetal bovine abomasum were investigated using transmission electron and light microscopic methods. In the oxyntic gland region oxyntic cell precursors and immature oxyntic cells appear as early as at the end of the first trimester of gestation--much earlier than described in any other mammalian animal species. Immature oxyntic cells are characterized by long apical microvilli, by their triangular-shaped light cytoplasm rich in large and numerous mitochondria, by the existence of vesicular profiles and by the incipient invagination of the apical plasma membrane forming a primitive intracellular canaliculus expanding into central areas of the cell. The oxyntic cell represents the first exocrine cell type developing from secretory granule-containing cells in the base of the primitive gastric glands.

Abomasum↗

Demonstration of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase in Drosophila Kc-cells.

Kc-cells from Drosophila melanogaster, grown under serum-free conditions, produce two beta-hexosaminidases and secrete these enzymes into the medium. The two enzymes were separated by DEAE-exchange chromatography. According to their substrate specificities one enzyme is a beta-N-acetyl-D-glucosaminidase (E.C.3.2.1.30), the other one a beta-N-acetyl-D-hexosaminidase (E.C.3.2.1.52). The beta-N-acetyl-D-glucosaminidase is predominant in the medium, the beta-N-acetyl-D-hexosaminidase within the cells. The Km values for the substrates pNP-GlcNAc, pNP-GalNAc, and (GlcNAc)2 are 0.8, 16.73, and 1.67 mM for the beta-N-acetyl-D-glucosaminidase and 0.24, 0.44, and 0.2 mM for the beta-N-acetyl-D-hexosaminidase. Both enzymes are inhibited by the products and the beta-N-acetyl-D-glucosaminidase is also inhibited stereospecifically by the substrates pNP-GlcNAc and (GlcNAc)2. Both enzymes are inhibited in a partial competitive way by acetamidolactones, the Kis being as low as 0.1 microM.

Acetylglucosaminidase↗

Physical properties of beta-N-acetyl-D-glucosaminidase and beta-N-acetyl-D-hexosaminidase from Drosophila Kc-cells.

Kc-cells from Drosophila produce two different beta-D-hexosaminidases, a beta-N-acetyl-D-glucosaminidase (E.C.3.2.1.30) and a beta-N-acetyl-D-hexosaminidase (E.C.3.2.1.52), which are also secreted into the medium. The Mr of both enzymes is about 126,000 +/- 9,700; the S-values are 8.37 +/- 0.44. Both enzymes have about the same pH optima at 5.5 and the same thermal stability. The temperature optima are identical (50 degrees C) for both enzymes if p-nitrophenyl-N-acetylglucosaminide is used as a substrate. However, when p-nitrophenyl-N-acetylgalactosaminide is used as the substrate the beta-N-acetyl-D-hexosaminidase has a temperature optimum about 10 degrees C higher. With higher salt concentrations, the activity of the beta-N-acetyl-D-glucosaminidase increases, whereas beta-N-acetyl-D-hexosaminidase is inhibited. Both enzymes also differ in their sensitivity to urea, the beta-N-acetyl-D-hexosaminidase being less sensitive than the beta-N-acetyl-D-glucosaminidase.

Acetylglucosaminidase↗