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Biomedical subjects

K Haglid

Publications and source records attributed to K Haglid.

At least 37 records · Page 2Linked to original sources

S-100 alpha-like immunoreactivity in tubules of rat kidney. A clue to the function of a "brain-specific" protein.

The localization of the alpha and beta subunits of S-100 protein was studied in normal tissue where the identification of three subclasses of S-100 containing cells was derived: i) cells that contain both alpha and beta subunits; ii) cells that contain only the alpha subunit; and iii) cells that contain only the beta subunit. In this study monospecific antibodies against the S-100 alpha and beta subunits were used to characterize the S-100-like immunoreactivity in the rat kidney: Certain cells in the distal nephron, i.e., the connecting piece, collecting ducts, and the thin limb of Henle's loop, contained S-100 alpha immunoreactivity. Proximal tubules, the thick ascending limb of Henle's loop, the distal tubules, and the juxtaglomerular apparatus were negative. No S-100 beta immunoreactivity was found in kidney tubules. However, Schwann cells of renal pelvic nerves contained S-100 beta immunoreactivity. The presence of S-100 alpha antigen in certain cells of the kidney gives further support to the assumption that the alpha subunit of S-100a is related to cells that are highly involved in pH, electrolyte, and water regulation.

Animals↗

Purification, characterization and ion binding properties of human brain S100b protein.

Human brain S100b (beta beta) protein was purified using zinc-dependent affinity chromatography on phenyl-Sepharose. The calcium- and zinc-binding properties of the protein were studied by flow dialysis technique and the protein conformation both in the metal-free form and in the presence of Ca2+ or Zn2+ was investigated with ultraviolet spectroscopy, sulfhydryl reactivity and interaction with a hydrophobic fluorescence probe 6-(p-toluidino)naphthalene-2-sulfonic acid (TNS). Flow dialysis measurements of Ca2+ binding to human brain S100b (beta beta) protein revealed six Ca2+-binding sites which we assumed to represent three for each beta monomer, characterized by the macroscopic association constants K1 = 0.44 X 10(5) M-1; K2 = 0.1 X 10(5) M-1 and K3 = 0.08 X 10(5) M-1. In the presence of 120 mM KCl, the affinity of the protein for calcium is drastically reduced. Zinc-binding studies on human S100b protein showed that the protein bound two zinc ions per beta monomer, with macroscopic constants K1 = 4.47 X 10(7) M-1 and K2 = 0.1 X 10(7) M-1. Most of the Zn2+-induced conformational changes occurred after the binding of two zinc ions per mole of S100b protein. These results differ significantly from those for bovine protein and cast doubt on the conservation of the S100 structure during evolution. When calcium binding was studied in the presence of zinc, we noted an increase in the affinity of the protein for calcium, K1 = 4.4 X 10(5) M-1; K2 = 0.57 X 10(5) M-1; K3 = 0.023 X 10(5) M-1. These results indicated that the Ca2+- and Zn2+-binding sites on S100b protein are different and suggest that Zn2+ may regulate Ca2+ binding by increasing the affinity of the protein for calcium.

Binding Sites↗

Dermal nerve sheath myxoma. A light and electron microscopic, histochemical and immunohistochemical study.

Two cases of dermal nerve sheath myxoma have been examined by light and electron microscopy, and by immunohistochemical and histochemical methods. The nomenclature of this tumor has been debated, and the histogenesis unsettled. Light microscopically, the lobulated myxoid tumors were generally composed of spindle-shaped and stellate tumor cells in an abundant matrix of mucosubstances. Ultrastructural features indicated an origin from the peripheral nerve sheath; among the pertinent findings were a single or duplicated external lamina investing the cells, desmosome-like junctions, cytoplasmic microfilaments and myelin figures, and interdigitating cytoplasmic processes. S-100 protein was demonstrated by means of the immunoperoxidase technique within both tumors, further supporting this origin. The histochemical analysis of the mucosubstance showed the presence of sulphated glucosaminoglycans. The name dermal nerve sheath myxoma, suggested by Harkin and Reed, is recommended.

Aged↗

Glial contribution to amino acid content and metabolism of the deafferented dentate gyrus.

The time course of tissue content and evoked release of endogenous amino acids was analyzed in the partially deafferented dentate gyrus of the rat hippocampus 2-24 days following unilateral lesion of the perforant path. Amino acids in tissue extracts and perfusates were determined after precolumn derivatization and hplc separation. The astrocytic glial cell reaction was monitored with immunohistochemistry of S-100. The tissue content of glutamate decreased significantly on the lesioned side, whereas only a moderate reduction in taurine, aspartate, and alanine occurred. Glutamine was significantly elevated at 7 days. The evoked efflux of glutamate was reduced at 2 and 7 days, whereas no change was seen at longer survival periods. The evoked release of GABA and aspartate increased on the denervated side after 12 and 24 days. The rate of carbon utilization into amino acid pools was followed with 14C-glucose and 14C-acetate. The incorporation of acetate showed a peak 2-9 days following lesion, which paralleled in time the hypertrophic glial cells. The incorporation of glucose decreased during this period. The metabolic events are discussed in relation to the morphological changes in synapses and glial cells.

Acetates↗

An immunohistochemical analysis of S-100 protein and glial fibrillary acidic protein in nasal glioma.

The authors describe a recurrent nasal glioma of the nasal septum in a new-born boy. Histologically, the nodular tumor tissue resembled normal glial tissue. Immunohistochemical studies with the immunoperoxidase (PAP and ABC) techniques revealed the presence of both S-100 protein and glial fibrillar acidic protein (GFAP), indicating the glial nature of the tumor.

Glial Fibrillary Acidic Protein↗

Differential localization of "brain-specific" S-100 and its subunits in rat salivary glands.

In the rat, the S-100 antigens in the submandibular gland were found to be immunochemically identical with those in the brain (glial cells) when compared using crossed immunoelectrophoresis. Specific antibodies against the S-100a non-beta and against the S-100 beta subunit were prepared from antibodies against crude S-100 protein and from S-100 components (S-100a and b) by affinity chromatography. In the rat salivary glands a differential distribution of subunit immunoreactivity was clearly evidenced using indirect immunofluorescence. Certain intercalated duct cells of the submandibular gland as well as Schwann cells contained the S-100 beta subunit immunoreactivity exclusively, while other duct cells in parotid, submandibular, and sublingual glands contained S-100a non-beta subunit immunoreactivity. Both subunits were present in astrocytes and ependymal cells. The immunocytochemical localization of alpha and beta subunits is a promising technique for the classification of various types of S-100-containing cells.

Animals↗

S-100 protein in melanocytic tumors. An immunohistochemical investigation of benign and malignant melanocytic tumors and metastases of malignant melanoma and a characterization of the antigen in comparison to human brain.

An immunohistochemical investigation regarding the presence of S-100 protein in benign and malignant, primary and metastatic melanocytic tumors is reported. The studied series consisted of 15 benign nevi, 3 blue nevi, 4 juvenile melanomas, 1 balloon cell nevus, 30 primary malignant melanomas of skin, mucous membranes and conjunctiva and 30 metastatic malignant melanomas. The immunohistochemical analysis showed positive staining for S-100 protein within the majority of the tumor cells in all benign tumors examined, except the balloon cell nevus, as well as in all the primary and metastatic malignant melanomas, including low-differentiated epitheloid or spindle-cell types without demonstrable melanin pigment. The results indicate that S-100 protein is a valuable marker for melanocytic tumors, especially in the fairly frequent dilemma of malignant melanoma presenting as a solitary non-pigmented metastasis of uncharacteristic light-microscopic appearance, and without known primary tumor. A characterization and quantification of the S-100 protein immunoreactivity in 5 metastatic malignant melanomas is presented. Using immuno-electrophoresis, the presence of 3 antigenic S-100 determinants was demonstrated within homogenates from the malignant melanomas, including the previously characterized S-100 A (alpha/beta) and S-100 B (beta/beta), and suggesting the presence of a hitherto undescribed variant of S-100 protein, possibly consisting of 2 alpha-subunits. Using rocket immuno-electrophoresis, the amount of S-100 protein was estimated.

Antigens, Neoplasm↗

The effect of S-100a and S-100b proteins and Zn2+ on the assembly of brain microtubule proteins in vitro.

The homologous proteins S-100a and S-100b affect the microtubule system in a distinctly different way in the presence of low molar ratios of Zn2+. Assembly of brain microtubule proteins can be almost completely inhibited and rapid disassembly can be induced by low molar amounts of S-100b in the presence of low molar ratios [2-4] of Zn2+. Higher molar ratios per S-100b (greater than 4) potentiate the general Zn2+ effect, promoting the formation of sheets of microtubules. However, the effect of S-100a is quite different, no inhibition of assembly can be observed and the presence of S-100a seems to protect the microtubule proteins against the effect of Zn2+ by chelating the Zn2+ and decreasing the free metal-ion concentration. S-100a or S-100b cannot bind to the microtubule polymer-form, either in the absence or in the presence of Zn2+.

Animals↗

Zinc ion binding to human brain calcium binding proteins, calmodulin and S100b protein.

Comparative studies have been performed on the binding properties of zinc ions to human brain calmodulin and S100b protein. Calmodulin is characterized by two sets of Zn2+ binding sites, with KD ranging from 8.10(-5)M to 3.10(-4)M. The S100b protein also exhibited two sets of zinc binding sites, with a much higher affinity. KD = 10(-7) - 10(-6)M. We suggest that S100b protein should no longer be considered only as a "calcium binding protein" but also as a "zinc binding protein", and that Zn2+ ions are involved in the functions of the S100 proteins.

Brain↗

Astrocytic development in fetal parietal cortex grafted to cerebral and cerebellar cortex of immature rats.

Pieces of cortex cerebri anlage were dissected out from 16- to 17-day-old fetuses and transplanted to the cortical and cerebellar regions of 5- to 6-day-old rat pups. Twelve animals with grafts in the cortical region and 5 animals with grafts in the cerebellar region were studied 1.5-4 months later. Cresyl violet stained sections revealed no gross difference in either cell morphology, cell density or cell distribution between grafts in the two locations. A molecular layer-like zone was present on all free surfaces of the grafts, whether facing a ventricle or the meninges. The astrocytic development was studied using immunohistochemistry with antibodies against glial fibrillary acidic protein, (GFA), and the S-100 protein. Both antibodies visualized starshaped astrocytes and perivascular membranes surrounding blood vessels. Semi-quantitative measurements as well as computerized image analysis showed that the total amount of GFA-like immunoreactivity was much higher in both types of grafts than in corresponding host cortex cerebri. No differences in amount of S-100-like immunoreactivity could be demonstrated. As S-100 is thought to be a more general astrocytic marker than GFA, this suggests that the difference in GFA-like immunoreactivity is due mainly to an increased amount of GFA within the individual astrocytes. It is concluded that grafts of fetal cortex cerebri pieces to the CNS of young hosts develop a profound astrocytic reaction characterized by an increased amount of GFA-like immunoreactivity.

Animals↗

Chronic exposure to trichloroethylene: lipid and acyl group composition in gerbil cerebral cortex and hippocampus.

Inhalation exposure of Mongolian gerbils to trichloroethylene (50 and 150 ppm) for 12 months caused only small changes in the lipid composition of their cerebral cortex and hippocampus. Total protein content and lipid class distribution were almost unaffected. However, the cholesterol to phospholipid ratio decreased in the cortex of animals exposed to trichloroethylene. This might be due to changes in the relative proportions of different membranes or cellular types. The finding of most interest was among the long-chain polyunsaturated fatty acids where changed ratios were found between long-chain derivatives of the linoleic and the linolenic acid series and a decrease in two minor fatty acids, 20:5 (n-3) and 22:5 (n-3). We think it is likely that such changes occur as a result of peroxidation of polyunsaturated fatty acids or in order to withstand the fluidizing properties of trichloroethylene.

Animals↗

Increased activity of a phospholipid base-exchange system by the differentiation of neoplastic cells from the nervous system.

The activity of a base-exchange enzyme system, specific for the polar head-groups of brain phospholipids, was measured in both C 1300 mouse neuroblastoma (clone 41A3) and human glioma 138 MG cell lines. This base-exchange is thought to occur between free nitrogenous bases and membrane-bound phospholipids. The incorporation of free 14C-labeled ethanolamine and L-serine was determined in undifferentiated and morphologically differentiated cells. Both cell lines showed a significant increase in ethanolamine and L-serine incorporation upon differentiation.

Animals↗

Partial purification of human kidney antigens.

A chromatographic method using immunoaffinity has been utilized in the search for kidney-specific antigens. After repeated absorptions by liver proteins coupled to a column of Sepharose CN-Br of a rabbit anti-human kidney serum, several antibodies remained. When these antibodies were coupled to a similar column and used for immunoaffinity chromatography, 8 kidney antigens, studied as peptides with molecular weights of 33,000, 80,000, 98,000 and 110,000-125,000 were obtained. They should not be found in similarly treated liver extracts.

Animals↗