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Biomedical subjects

M Betz

Publications and source records attributed to M Betz.

At least 37 records · Page 2Linked to original sources
PubMed1991

Cholesterol-transfer protein located in the intestinal brush-border membrane. Partial purification and characterization.

Cholesterol absorption by small intestinal brush border membrane vesicles from taurocholate mixed micelles is a second-order reaction. From a comparison of reaction rates and order before and after proteinase K treatment of brush-border membrane vesicles, it is concluded that cholesterol absorption is protein-mediated. It is shown that the desorption of cholesterol from taurocholate mixed micelles is by a factor of about 10(4) faster than that from egg phosphatidylcholine bilayers. When brush border membrane vesicles are stored at room temperature, intrinsic proteinases are activated and proteins are liberated from the brush border membrane. These proteins collected in the supernatant catalyze cholesterol and phosphatidylcholine exchange between two populations of small unilamellar phospholipid vesicles. One of the active proteins present in the supernatant is purified by a two-step procedure involving gel filtration on Sephadex G-75 SF and affinity chromatography on a Nucleosil-phosphatidylcholine column. The protein thus obtained is pure by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. It has an apparent molecular weight of slightly less than 14,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and a value of 11,500 determined by gel filtration on Sephadex G-75 SF.

Animals↗
PubMed1991

Prostaglandin E2 inhibits production of Th1 lymphokines but not of Th2 lymphokines.

PGE2 is known to inhibit IL-2 and IFN-gamma production from Th cells and is widely viewed as a general immunosuppressant. However, PGE2 was found not to inhibit IL-4 production from Th2 clones, and IL-5 production from these clones was slightly enhanced. The same results were obtained with short term T cell lines, which indicates that the lack of inhibition of IL-4 and IL-5 production by PGE2 is a general phenomenon. PGE2 functions by increasing cAMP levels through activation of adenylate cyclase. Despite its failure to inhibit lymphokine release, PGE2 was capable of increasing cAMP levels in Th2 cells, and forskolin, a direct activator of adenylate cyclase, also did not inhibit IL-4 or IL-5 production. These data indicate that the failure of PGE2 to inhibit IL-4 and IL-5 production was not due to an inability of PGE2 to induce an increase in intracellular cAMP, and suggested instead that the expression of IL-4 and IL-5 in Th2 cells is insensitive to elevated cAMP levels. When Th0 clones were examined, PGE2 was again found to differentially affect IL-2 and IL-4 production in three of five clones tested. In two additional Th0 clones, both IL-2 and IL-4 production were inhibited. These data suggest that lymphokine production may be regulated on two different levels. First, Th1- and Th2-associated lymphokines may be differentially sensitive to intracellular signals such as cAMP. Second, T cell subsets may exist, including subsets of Th0 cells, with different signaling pathways. In addition, our data suggest that PGE2 may play an important role in regulating the development of a response dominated by Th1- or Th2-associated lymphokines.

Animals↗
PubMed1991

Stimulation of beta 2-microglobulin synthesis in lymphocytes after exposure to Cuprophan dialyzer membranes.

We tested beta 2-microglobulin (beta 2m) synthesis by peripheral blood lymphocytes of non-uremic donors after contact with dialyzer membranes. Under serum-free conditions, lymphocytes were incubated with different dialyzer membranes for four hours. After subsequent culture in the absence of membranes to yield an overall culture time of 24 hours, RNA was extracted. Messenger RNA for beta 2m was quantitated by Northern blotting with specific probes. Increased beta 2m mRNA was seen after a minimum of one hour contact time with Cuprophan both in the presence and absence of cycloheximide. While no consistent stimulation was seen with polyacrylonitrile or polycarbonate-polyether membranes, respectively, the stimulation of beta 2m mRNA with Cuprophan was of the same order of magnitude as that obtained with interferon gamma. Stimulation of beta 2m mRNA by Cuprophan was stereospecifically diminished by 5 mM L-fucose. Apart from stimulated transcription of beta 2m we could also show increased surface expression of MHC class I molecules, using FACS technique. The results further prove complement-independent blood cell activation by Cuprophan membranes. It is uncertain whether the results are relevant for beta 2m amyloidosis.

Blood Proteins↗
PubMed1991

Immunological characterization of two dominant tonoplast polypeptides.

At least 14 distinct polypeptides reside in the tonoplast of barley (Hordeum vulgare) mesophyll vacuoles. Two of the polypeptides were isolated from two-dimensional separations of vacuoplast membrane proteins and used for immunization. With the antisera, the localization on the membrane and the distribution of the polypeptides in the plant kingdom and in various tissues of barley plants was studied. The polypeptides have an apparent molecular mass of 31 and 40 kilodaltons. After freeze-thaw cycles or washing of the membranes with 4.5 millimolar NaCl, the polypeptides were still sedimented with the membranes, suggesting an intrinsic localization. The antiserum against the 31-kilodalton polypeptide bound to the outer surface of isolated intact vacuoles. In chromatographic separations of Triton X-100-solubilized membrane fractions, the residual activities of various acid hydrolases eluted distinct from the 31- and 40-kilodalton polypeptides. Both polypeptides tend to form larger aggregates, however smaller than the tonoplast ATPase. Cross-reactive polypeptides were present in higher and lower plants (the green alga Chara corallina and the liverwort Conocephalum) and in liver tissue from rat and beef, but were not detected in other animal tissues tested so far. The results indicate a wide distribution of these tonoplast polypeptides in vacuole-containing organisms.

Journal Article↗
PubMed1990

Regulation and development of cytochrome c-specific IL-4-producing T cells.

The development of Ag-specific IL-4-producing T cells in short term cultures was examined. Freshly harvested lymph node cells from B10.A mice immunized with the cytochrome c fragment 81-104 did not produce detectable amounts of IL-4 upon Ag stimulation. However, after one cycle of bulk in vitro restimulation, the same cells could be stimulated to secrete IL-4. The development of Ag-specific IL-4-producing cells during the in vitro culture could be influenced by different culture conditions. When IL-4 and IL-1, in the presence of anti-IL-2R antibodies, were added to the bulk culture, secondary stimulation of the cells revealed increased levels of IL-4 production and constant or decreased levels of IL-2 production. The addition of IFN-gamma during the bulk culture led to a decrease in IL-4 production, yet had no effect on IL-2 production. When anti-IL-4 antibodies were present during the restimulation culture, no IL-4 was produced in the final stimulation assay. In addition, T cells cultured at high cell density produced more IL-4 in a secondary stimulation than did T cells cultured at low cell density. These results demonstrate that culture conditions during a short term culture of freshly harvested primed T cells may profoundly influence the development of IL-4-producing cells. This may be caused either by selective expansion or inhibition of preexisting IL-4-producing T cells, or by the differentiation of precursors of IL-4-producing cells.

Animals↗
PubMed1988

Nystatin stimulates prostaglandin E synthesis and formation of diacylglycerol in human monocytes.

The effect of the channel forming antibiotic nystatin on human monocytes was studied. Monocytes isolated from the peripheral blood (Mo), the monocytic cell line U937 and the promyelocytic cell line HL60 were used. Each cell type could be lysed with nystatin. The dose of nystatin required, however, was different for each cell line. In sublytic doses nystatin induced a rise of intracellular Ca2+, measured with the calcium indicator quin2. The rise of intracellular Ca2+ was followed by the release of prostaglandin E. By preincubation of the cells with quin2 the prostanoid synthesis could be inhibited suggesting that the increased Ca2+-levels could function as a signal. The prostanoid synthesis was also suppressed by inhibitors of the arachidonic acid pathway. Furthermore, nystatin induced an increase of diacylglycerol and a decrease of phosphatidylinositol. The generation of diacylglycerol, however, was not due to hydrolysis of the polyphosphoinositides because no increase of the second messenger inositol-1,4,5-triphosphate could be detected.

Calcimycin↗
PubMed1988

Cuprammonium membranes stimulate interleukin 1 release and arachidonic acid metabolism in monocytes in the absence of complement.

Contact of blood with foreign surfaces, specifically dialysis membranes, causes cell activation which has widely been assumed to be mediated by complement (C). To explore the possibility of C-independent activation, we examined different cell types: PMN, human monocytes and the cell lines U937 or HL60, washed human platelets and rat glomerular epithelial cell (primary) cultures (GEC), under serum-free conditions and after addition of anti-C3 F(ab)2, respectively. The monitored biological effects were release of PGE2, TXB2 or interleukin 1 and generation of O2- radicals. To further explore the mechanisms involved, phospholipid metabolism was studied by measuring IP3 and DG (14C-arachidonic or oleic acid prelabeled U937 and HL60 cells); changes of cytosolic Ca++ (Quin2 technique) were also determined. The results show that in absence of C, brief (2 min) contact with cuprammonium (CU) stimulated: (a) PGE2 release in U937 and human monocytes or GEC; (b) TXB2 release in washed platelets; (c) slow interleukin 1 release by monocytes; and (d) generation of O2- radicals in PMN. Artifacts due to endotoxin were excluded by appropriate polymyxin control experiments and by comparison of effects with those of bacterial LPS. Potential synthesis of C3 by U937 was excluded by addition of anti-C3 F(ab)2. C-independent cell activation was accompanied by increase of DG, but not IP3 (suggesting involvement of protein C kinase dependent mechanisms) and by increased cytosolic Ca++. To further explore the initial signal involved, incubations were carried out with covalently modified CU members (DEAE cellulose) and in the presence of mM concentrations of monosaccharides. Cationic modification of CU membranes reduced C-independent cell activation.(ABSTRACT TRUNCATED AT 250 WORDS)

Arachidonic Acid↗
PubMed1988

The complement membrane attack complex stimulates the prostanoid production of cultured glomerular epithelial cells.

Incubation of cultured rat glomerular epithelial cells (GEC) with sublytic amounts of the purified complement components C5b6, C7, C8 and C9 greatly stimulated the release of the prostanoids prostaglandin E (PGE) and thromboxane B2. Incubation of GEC with C5b-8 was also stimulatory, whereas omission of C7 abolished the enhanced prostanoid production. These effects were dose-dependent. The increased release of PGE was biphasic with peaks at 5 min and 24 h of incubation. The second peak could be prevented by treatment with cycloheximide, suggesting its dependence on protein synthesis. The observations on cultured GEC provide evidence that terminal complement components alter the metabolism of glomerular cells, resulting in increased production of prostanoids. The results are consistent with the concept that deposition of nonlytic amounts of complement in the glomerular capillary wall may affect the GEC in vivo and may indirectly contribute to abnormalities of the glomerular filter as it is seen in glomerular disease.

Animals↗
PubMed1987

Thromboxane B2 synthesis in human platelets induced by the late complement components C5b-9.

Sublytic doses of purified C5b6, C7, C8 and C9 induce the release of thromboxane B2 (TXB2) in human platelets. In the present study, we attempted to analyze the signal by which C5b-9 triggers the prostanoid synthesis in platelets. TXB2 release was accompanied by liberation of Ca++ from intracellular stores. Influx of Ca++ was not observed, indicating that C5b-9 did not form a channel large enough to permit passage of Ca++ ions. Therefore, channel formation is apparently not required for cell stimulation.

Arachidonic Acid↗
PubMed1987

Effect of the late complement components C5b-9 on human monocytes: release of prostanoids, oxygen radicals and of a factor inducing cell proliferation.

Recently, we reported stimulation of rat macrophages and human platelets by isolated C5b-9 to synthesize prostaglandin E (PGE) or thromboxane B2 (TXB2). In the present study, we tested whether besides prostanoids, C5b-9 also would induce the production of other mediators. We found that C5b-9 in sublytic concentrations stimulated human granulocytes (polymorphonuclear leukocytes) or monocytes to release oxygen radicals. Furthermore, monocytes release interleukin-1 in response to C5b-9. Thus, besides having a lytic capacity, C5b-9 also functions as a stimulator of various cells.

Complement Membrane Attack Complex↗
PubMed1986

Control of echolocation pulses by neurons of the nucleus ambiguus in the rufous horseshoe bat, Rhinolophus rouxi. I. Single unit recordings in the ventral motor nucleus of the laryngeal nerves in spontaneously vocalizing bats.

The vocal motor control of the larynx was studied with single unit recordings from the efferent motor nucleus (nucleus ambiguus) in the CF-FM-bat Rhinolophus rouxi, spontaneously emitting echolocation sounds. The experiments were performed in a stereotaxic apparatus that allowed differentiation of activities in the recorded nucleus depending on the electrode position (Fig. 1). Echolocation calls and respiration activity were monitored simultaneously, thus it was possible to compare the time course of the motor control activity during respiration with and without concurrent vocalization. Unit discharges were classified as laryngeal motoneuron activity according to their correlation with the time course (onset and end) of echolocation calls and their discharge rate as: Pre-off-tonic, pre-off-phasic, off-pauser, off-tonic, on-chopper, on-tonic, prior-tonic and inhibitory (Fig. 4). The on-chopper and on-tonic discharge patterns were assigned to the motor activity of the lateral cricoarytenoid muscle and the off-pauser and off-tonic discharge patterns to the motor activity of the posterior cricoarytenoid muscle controlling the time course of vocal pulses. Motoneuron activities recorded under the condition of systematically shifted frequencies in the emitted echolocation calls were investigated in Doppler-shift compensating bats responding to electronically simulated echoes. Of all neurons classified as motor control, only units of the pre-off-tonic discharge type (cricothyroid muscle) changed their activity with frequency shifts in the vocalized pulses; they showed a positive linear correlation with the emitted sound frequency (Fig. 6). In addition, single unit activities in strict synchronization to vocalization were recorded, that by their low discharge rate were not valid as motor control, and were considered to represent activities of interneurons or internuclear neurons connecting the nucleus ambiguus with other vocalization- and respiration-centers (Fig. 3c). Electric lesions in the brain stem and iontophoretically applied horseradish peroxidase (HRP) served as references for localization and morphological identification of the recording sites in cell stained brain slices.

Animals↗
PubMed1986

A stereotaxic method for small animals using experimentally determined reference profiles.

In bats conventional stereotaxic methods do not yield sufficient positional accuracy to allow reliable recordings and tracer injections in subnuclei of the auditory system. In a newly developed stereotaxic system experimentally measured patterns of skull profile lines are used to define the animal's brain position with an accuracy of +/- 100 microns. By combining the neurophysiological stereotaxic procedure with a standardization of the neuroanatomical processing of the brains, the location of recordings, stimulations or injections can be readily transformed into brain atlas coordinates. This facilitates the compilation and comparison of data within and among animals. The system is not restricted to use in bats and can be readily adapted to other experimental animals.

Animals↗
PubMed1985

An HRP-study of the frequency-place map of the horseshoe bat cochlea: morphological correlates of the sharp tuning to a narrow frequency band.

The frequency-place map of the horseshoe bat cochlea was studied with the horseradish peroxidase (HRP) technique involving focal injections into various, physiologically defined regions of cochlear nucleus (CN). The locations of labeled spiral ganglion cells and their termination sites on inner hair cells of the organ of Corti from injections into CN-regions responsive to different frequencies were analyzed in three dimensional reconstructions of the cochlea. Horseshoe bats from different geographical populations were investigated. They emit orientation cells with constant frequency (CF) components around 77 kHz (Rhinolophus rouxi from Ceylon) and 84 kHz (Rhinolophus rouxi from India) and their auditory systems are sharply tuned to the respective CF-components. The HRP-map shows that in both populations: the frequency range around the CF-component of the echolocation signal is processed in the second half-turn of the cochlea, where basilar membrane (BM) is not thickened, secondary spiral lamina (LSS) is still present and innervation density is maximal; frequencies more than 5 kHz above the CF-component are processed in the first half-turn, where the thickened BM is accompanied by LSS and innervation density is low; frequencies below the spectral content of the orientation call are represented in apical turns showing no morphological specializations. The data demonstrate that the cochlea of horseshoe bats is normalized to the frequency of the individual specific CF-component of the echolocation call. The HRP-map can account for the overrepresentation of neurons sharply tuned to the CF-signal found in the central auditory system. A comparison of the HRP-map with a map derived with the 'swollen nuclei technique' following loud sound exposure (Bruns 1976b) reveals that the latter is shifted towards cochlear base by about 4 mm. This discrepancy warrants a new interpretation of the functional role of specialized morphological structures of the cochlea within the mechanisms giving rise to the exceptionally high frequency selectivity of the auditory system.

Animals↗
PubMed1984

Ehrlich ascites cells activate the alternative pathway of the human complement system.

Incubation of Ehrlich ascites cells with normal or C1q or C2 deficient human sera results in killing of the cells. Killing occurred also in the absence of free Ca++, which supported by the fact that factor B and C3 were cleaved, leads to the conclusion that the alternative pathway of the complement system is activated on the surface of the Ehrlich ascites cells.

Animals↗
PubMed1984

Release of arachidonic acid: a new function of the late complement components.

Incubation of Ehrlich-ascites cells with sublytic complement doses results in degradation of membrane phospholipids and release of arachidonic acid. Phospholipase A2 blockers inhibit arachidonic acid release indicating a phospholipase dependent cleavage of the phospholipids. Phospholipase A2 is apparently activated during complement-membrane interaction, representing a new reactivity of the late complement components.

Animals↗
PubMed1984

Macrophages release arachidonic acid, prostaglandin E2, and thromboxane in response to late complement components.

Rat peritoneal macrophages released arachidonic acid, prostaglandin E2, and thromboxane B2 when treated with normal rabbit or C6-deficient rabbit complement in vitro. Normal rabbit complement, however, was more efficient, which indicates that late complement components, in addition to the known effects of C3a and C3b, were responsible for an enhanced arachidonic acid turnover. Indeed, in the absence of the C3 cleavage products, the purified late complement components C5b6, C7, C8, and C9 stimulated the arachidonic acid, as well as the prostaglandin E2 and thromboxane B2 release. Incubation of C5b6, C7, C8, and C9 for 1 hr at 37 degrees C before addition to the macrophages abolished the stimulatory activity, being in complete agreement with the fact that a fluid phase-formed complex of C5-9 loses its membrane-binding capacity. Although the mechanism by which C5b-9-membrane interaction affects the arachidonic acid metabolism remains unclear, the data describe a new function of the late complement components.

Animals↗