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Biomedical subjects

K Keller

Publications and source records attributed to K Keller.

At least 73 records · Page 4Linked to original sources
PubMed1994

The use of proliferating cell nuclear antigen (PCNA) expression and intraepithelial lymphocyte count in the differential diagnosis of inflammatory bowel disease.

Colonic biopsies of 283 patients with Crohn's disease (CD), ulcerative colitis (UC), infectious colitis (IC) lymphocytic colitis (LC) and controls for patients with either CD or UC, matched for age and sex, were evaluated with respect to the number of intraepithelial lymphocytes (IEL) and PCNA (cyclin) positive crypt epithelia. The IEL count of patients with LC differed significantly from all other patient groups, indicating that this condition bears no relation to the other diseases. Our patients with histologic diagnosis of LC had no uniform clinical presentation and the significance of this characteristic increase in IEL is unclear. The mean value of PCNA positive crypt epithelia in the lower crypt half was significantly higher in UC patients than in CD patients. PCNA counts in patients with IC or LC were not different from values in UC patients. PCNA counts in CD patients overlapped frequently with those in UC patients. This parameter thus fails to distinguish between CD and UC in individual patients and, though not investigated by us, is unlikely to be effective in the differential diagnosis in patients with a provisional diagnosis of indeterminate colitis.

Adult↗
PubMed1994

[A procedure for assessment "relaxed state" with reference to basic exercises in autogenic training].

The starting point of the present article was the fact that there is practically never a baseline-testing of "relaxability" before a relaxation training starts. This article presents such a procedure, which has been adapted from the "Autogenic Training". The procedure is suitable for application to groups, and consists of an instruction text for 11 exercises and a questionnaire. The psychometric properties of the questionnaire were checked with data of an accidental sample (n = 106) of an orthopedic rehabilitation clinic. In order to check for the stability over time, the procedure was repeated after an interval of two to three weeks. Cronbach's coefficient alpha was .96 and .97 for the first and second testing, respectively. The corrected item-total-correlation varied between .56 and .87 for both testings. The retest-reliability of the score was .75. Factor-analysis yielded two factors for each testing, respectively. For one factor, items mainly referring to the heaviness- and warmthexercises had highest loadings. Items referring to the gerenal relaxation predominantly belonged to the other factor. Consequently, all the analyses point to the stability of the results over time.

Adult↗
PubMed1993

GTP analogs suppress uptake but not transport of D-glucose analogs in Glut1 glucose transporter-expressing Xenopus oocytes.

A Xenopus oocyte expression-co-injection system was used to study the influence of guanine nucleotides on D-glucose uptake. GTP analogs like GTP gamma S and GppNHp had no effect on 3-O-methylglucose transport determined by zero-trans uptake or equilibrium exchange, but suppressed 2-deoxyglucose uptake into Glut1 glucose transporter-expressing oocytes by up to 86%. Both GTP analogs showed concentration dependence of their effectiveness, with GTP gamma S being more potent than GppNHp. No statistically significant differences were observed between groups of oocytes co-injected with water or GDP beta S (250 and 500 microM intracellular concentration). Glut1 transporter expression in plasma membrane was not different between water or GTP gamma S-co-injected oocytes. Thus, inhibition of hexokinase catalytic activity is the most likely causative factor for down-regulation of 2-deoxyglucose uptake.

Animals↗
PubMed1993

Glucose transport activity and photolabelling with 3-[125I]iodo-4-azidophenethylamido-7-O-succinyldeacetyl (IAPS)-forskolin of two mutants at tryptophan-388 and -412 of the glucose transporter GLUT1: dissociation of the binding domains of forskolin and glucose.

The tryptophan residues 388 and 412 in the glucose transporter GLUT1 were altered to leucine (L) by site-directed mutagenesis and were transiently expressed in COS-7 cells. As assessed by immunoblotting, comparable numbers of glucose transporters were present in plasma membranes from cells transfected with wild-type GLUT1, GLUT1-L388 or GLUT1-L412. Transfection of the wild-type GLUT1 gave rise to a 3-fold increase in the reconstituted glucose transport activity recovered from plasma membranes. In contrast, transfection of GLUT1-L412 failed to increase the reconstituted transport activity, whereas transfection of GLUT1-L388 produced only a 70% increase. Photolabelling of GLUT1-L412 with 3-[125I]iodo-4-azidophenethylamido-7-O-succinyldeacetyl (125IAPS)-forskolin was not different from that of the wild-type GLUT1, whereas the GLUT1-L388 incorporated 70% less photolabel than did the wild-type GLUT1. These data suggest a dissociation of the binding sites of forskolin and glucose in GLUT1. Whereas both tryptophan-388 and tryptophan-412 appear indispensable for the function of the transporter, only tryptophan-388 is involved in the binding of the inhibitory ligand forskolin.

Affinity Labels↗
PubMed1993

Endocarditis caused by a Leptotrichia buccalis-like bacterium in a patient with a prosthetic aortic valve.

The isolation of a Leptotrichia buccalis-like bacterium from the blood of a patient with endocarditis associated with a prosthetic aortic valve is reported. The organism was isolated from different blood cultures. It was similar but not completely identical to Leptotrichia buccalis. The microbiological identification was confirmed independently by three laboratories, including two reference institutions. The case and the characteristics of the microorganism are reported.

Aged↗
PubMed1993

Study of the membrane permeability of a paramagnetic metal complex on single cells by NMR microscopy.

A new procedure has been developed for investigating the ability of paramagnetic metal complexes to penetrate the plasma membrane of eukaryotic cells without decomposition. Defolliculated Xenopus laevis oocytes formed the biological system to test N,N-ethylenebis-(1,5,5-trimethyltetramic-acid-3-acetiminato) copper (II). An increase of the signal intensities in spin-echo (SE) images of oocytes treated with the tested substance indicated that the complex was able to penetrate biological membranes due to the arrangement of hydrophobic and hydrophilic groups within the ligand. In contrast, the treatment with the commonly used contrast agent gadolinium-DTPA/dimeglumine did not enhance the signal intensity in NMR images of oocytes after time periods of exposure comparable to those used for the copper complex. After microinjection into Xenopus oocytes the copper complex was released into the extracellular medium without degradation, as shown by HPLC measurements.

Animals↗
PubMed1993

Entamoeba histolytica proteins modulate the respiratory burst potential by murine macrophages.

Reactive oxygen intermediates are important components of macrophage microbicidal mechanisms and pathogenesis of parasitic disease. The purpose of the present study was to investigate the effect of virulent Entamoeba histolytica (strain HM1-IMSS) on respiratory burst potential of macrophages. Pretreatment of elicited peritoneal macrophages (EPM) with crude soluble amoebic proteins from 1 to 6 hr was found to prime EPM for enhanced O2 and H2O2 release in response to phorbol myristate acetate (PMA) in a dose-dependent manner, whereas pretreatment with the same concentrations of the non-pathogenic E. histolytica-like Laredo strain was without priming effect. Low molecular weight (MW) amoebic proteins (27,000-67,000) purified by Sephacryl-200 column chromatography and subfractionated by diethylaminoethyl cellulose chromatography were 10-fold more potent than crude amoebic proteins in priming EPM for an enhanced respiratory burst potential. Both crude and purified amoebic proteins inhibited the priming effect of lipopolysaccharide (LPS) or interferon-gamma (IFN-gamma) and antagonized the stimulating effect of PMA. Amoebic proteins by themselves were incapable of stimulating EPM respiratory burst. These findings demonstrate that amoebic proteins are capable of modulating the respiratory burst response of macrophages, suggesting an important role for them in the immunoregulation and pathogenesis of amoebiasis.

Animals↗
PubMed1992

The differential role of Cys-421 and Cys-429 of the Glut1 glucose transporter in transport inhibition by p-chloromercuribenzenesulfonic acid (pCMBS) or cytochalasin B (CB).

Cys-421 and Cys-429 of Glut1 were replaced by site-directed mutagenesis in order to investigate their involvement in basal glucose transport and transport inhibition. Neither of the two cysteine residues was essential for basal 2-deoxy-D-glucose uptake in Xenopus oocytes expressing the respective mutant M421 and M429. If applied from the external side, the poorly permeable sulfhydryl-reactive agent pCMBS inhibited 2-deoxy-D-glucose uptake of Glut1- and M421-expressing Xenopus oocytes but failed to affect uptake of the Cys-429 mutant. This is in agreement with the proposed two-dimensional model of Glut1 confirming that Cys-429 is the only residue exposed to the surface of the plasma membrane. The replacement of Cys-421 at the exofacial end of helix eleven caused a partial protection of 3-O-methylglucose transport inhibition by CB; this residue may thus be involved in stabilizing an adjacent local tertiary structure necessary for the full activity of this inhibitor.

4-Chloromercuribenzenesulfonate↗
PubMed1992

Subcellular distribution and activity of glucose transporter isoforms GLUT1 and GLUT4 transiently expressed in COS-7 cells.

In adipose and muscle cells, the glucose transporter isoform GLUT4 is mainly located in an intracellular, vesicular compartment from which it is translocated to the plasma membrane in response to insulin. In order to test the hypothesis that this preferential targeting of a glucose transporter to an intracellular storage site is conferred only by its primary sequence, we compared the subcellular distribution of the fat/muscle glucose transporter GLUT4 with that of the erythrocyte/brain-type glucose transporter GLUT1 after transient expression in COS-7 cells. Full-length cDNA was ligated into the expression vector pCMV that is driven by the cytomegalovirus promoter, and introduced into COS cells by the DEAE-dextran method. Cells were homogenized and fractionated by differential centrifugation to yield plasma membranes and a Golgi-enriched fraction of intracellular membranes (low-density microsomes). In these membrane fractions, the abundance of glucose transporters was assessed by immunoblotting with specific antibodies against GLUT1 and GLUT4, and their transport activity was assayed after solubilization and reconstitution into lecithin liposomes. Uptake rates of 2-deoxyglucose assayed in parallel samples were higher in cells expressing GLUT1 or GLUT4 as compared with control cells (transfection of pCMV without transporter cDNA). Reconstituted glucose transport activity in plasma membranes was about 5-fold higher after expression of GLUT1 and GLUT4 as compared with control cells. The relative amount of GLUT4 in the low-density microsomes as detected by reconstitution and immunoblotting exceeded that of the GLUT1, but was much lower than that observed in typical insulin-sensitive cells, e.g., rat fat cells or 3T3-L1 adipocytes. These data indicate that COS-7 cells transfected with glucose transporter cDNA express the active transport proteins and can be used for functional studies.

Amino Acid Sequence↗
PubMed1992

Amino acid substitutions at tryptophan 388 and tryptophan 412 of the HepG2 (Glut1) glucose transporter inhibit transport activity and targeting to the plasma membrane in Xenopus oocytes.

All 6 tryptophan residues in the human HepG2-type glucose transporter (Glut1) were individually altered by site-directed mutagenesis to investigate the role of these residues in transport function. Tryptophan residues in positions 48, 65, 186, 363, 388, and 412 of Glut1 were changed to either a glycine or leucine residue. Mutant mRNAs were synthesized and injected into Xenopus laevis oocytes. Transporter function as assessed by uptake of 2-deoxy-D-[3H]glucose or transport of 3-O-[3H]methylglucose was decreased in the 388 and 412 mutants but was unaltered in all other mutants. The amount of the mutant transporters expressed in total membrane and plasma membrane fractions was measured using Glut1-specific antibodies. Calculation of the intrinsic transport activity of each of the mutants using these data demonstrated that the reduced transport activity of the 412 mutants was caused entirely by a dramatic decrease in the intrinsic activity of the mutant proteins whereas the reduced activity of the 388 mutants was a result of a decreased level of the protein in oocytes, decreased targeting to the plasma membrane, and a modest decrease in the intrinsic activity. Protease/glycosidase mapping of in vitro translation products indicated that the effects of the 388 and 412 point mutations could not be attributed to a disruption in the ability of the mutant proteins to insert properly into the membrane. The ID50 for cytochalasin B inhibition of 2-deoxyglucose uptake was increased from 5 x 10(-7) M for the wild-type Glut1 to 4 x 10(-6) M in the 388 mutants but was unaltered in the 412 mutants. These observations suggest that 1) Trp-412 may comprise part of a hexose binding site or is involved in maintaining a local tertiary structure critical for transport function; 2) Trp-388 is involved in stabilizing the equilibrium binding of cytochalasin B to the transporter. Trp-388 may therefore lie near a substrate binding site and also appears to participate in stabilization of local tertiary structure important for full catalytic activity and efficient targeting to the Xenopus plasma membrane.

Animals↗
PubMed1992

Modulation of tumor necrosis factor production by macrophages in Entamoeba histolytica infection.

The macrophage-derived mediator tumor necrosis factor alpha (TNF) is a cytokine with pleiotropic effects. TNF exhibits potent immunologic and inflammatory properties in parasitic diseases. The present study examined the production of TNF by macrophages isolated from gerbils infected with Entamoeba histolytica and by naive macrophages in response to amoebae in vitro. Amoebic liver abscess-derived macrophages produced low constitutive basal levels of TNF; in response to lipopolysaccharide (LPS) stimulation, TNF production was enhanced by 14-, 11-, and 6-fold at 10, 20, and 30 days postinfection, respectively. Amoebic liver abscess-derived macrophages pretreated with either recombinant gamma interferon (IFN-gamma) or the cyclooxygenase inhibitor indomethacin augmented TNF production in response to soluble amoebic proteins and LPS. Kupffer cells and peritoneal and spleen macrophages from infected animals did not release TNF constitutively in vitro. However, TNF production in response to LPS stimulation was significantly higher at 10 and 20 days postinfection. Macrophages from infected and naive animals pretreated with recombinant IFN-gamma or indomethacin produced increased amounts of TNF in response to LPS but not in response to soluble amoebic protein stimulation. Pretreatment of naive macrophages with amoebic proteins inhibited LPS-induced TNF production by 69 to 79%; the effect of the amoebic proteins was partially reversed by indomethacin pretreatment. In contrast, IFN-gamma- and LPS-activated naive macrophages produced enhanced levels of TNF in response to live amoebae and soluble amoebic proteins. Our results demonstrate that TNF production by macrophages is altered during E. histolytica infection and in response to amoebae and suggest a role for IFN-gamma and prostaglandin E2 in regulating TNF production during the infection.

Animals↗
PubMed1992

The fast release of mucin secretion from human colonic cells induced by Entamoeba histolytica is dependent on contact and protein kinase C activation.

The mucus producing colonic cell line, LS174T, was used as a model to study E. histolytica-induced mucin secretion. E. histolytica trophozoites in contact with the mucus layer overlying the LS174T cells and in response to PMA, a protein kinase C activator, and Ca2+ ionophore A23187 which elevates intracellular Ca2+ ([Ca]i), caused a time-dependent (0.25-2.00 h) release of mucin. PKC inhibitors, H7 and staurosporine inhibited E. histolytica (37 and 75%) and PMA (46 and 100%)-induced mucin secretion, whereas in response to Ca2+ ionophore mucin secretion was augmented (56 and 17%). Both PMA and E. histolytica-induced the translocation of the PKC enzyme from the cytoplasm to the membrane fraction with increased enzyme activity. These results suggest that even though mucin secretion can be induced by PKC and Ca(2+)-dependent pathways, E. histolytica evokes the fast release of mucins by a PKC-dependent mechanism.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

[The legal status of homeopathic medicines. Quality requirements, registration and permit procedures].

Homeopathic medicaments which are ready for application have to be approved or registered before they can be made available. This procedure is regulated by the "Arzneimittelgesetz (AMG)", a law regulating licensing and registration of medicaments. The commission in charge of the registration and licensing of homeopathic medicaments is the "Aufbereitungskommission D" of the German "Bundesgesundheitsamt (BGA)", the Board of Public Health. Application forms are dealt with on the basis of controlled and non-controlled clinical studies, application observations, and case studies, as well as specialized literature. The commission is trying to do justice to the particular features of the special field of homeopathy.

Animals↗
PubMed1991

Down-regulation of murine lymphocyte responsiveness to mitogens after treatment with antigens of Entamoeba histolytica.

Injection of mice with pathogenic Entamoeba histolytica (strain HM1-IMSS) antigens resulted in a decreased capacity of splenocytes to respond to mitogen-induced blastogenesis following a challenge with concanavalin A (Con A), phytohemagglutinin (PHA), and lipopolysaccharide (LPS), whereas no inhibition was observed in mice that had previously been injected with equivalent amounts of non-pathogenic E. histolytica-like Laredo antigens. Depletion of adherent cells in the splenocyte preparation indicated that these cells were not a major contributor to the observed immunosuppression. Quantification of splenic T-lymphocyte subsets demonstrated a significant decrease in Thy-1+ and Lyt-1+ cells, but Lyt-2+ cells were not affected. Splenocytes treated with pathogenic amoebic antigens in vitro affected the capacity of these cells to respond optimally to Con A- and LPS-induced blastogenesis but not to that induced by PHA. These findings demonstrate that amoebic antigens affect lymphocyte function and may be important co-factors in the immunoregulation and pathogenesis of amoebiasis.

Animals↗
PubMed1991

Mucin and nonmucin secretagogue activity of Entamoeba histolytica and cholera toxin in rat colon.

Depletion of colonic mucus occurs before invasion of the colonic mucosa by Entamoeba histolytica trophozoites. It is hypothesized that E. histolytica releases a mucus secretagogue; this was studied in a rat colonic loop model. In colonic loops exposed to live amebae, mucus secretion was quantitated by release of acid-precipitable [3H]glucosamine-labeled luminal glycoprotein and by specific immunoassay. Mucus secretion increased in dose-dependent fashion in response to greater than or equal to 1 X 10(5) trophozoites; cholera toxin (20 micrograms per loop), a known mucus secretagogue, elicited a similar response. Thin-section histological analysis of amebae and cholera toxin-exposed loops showed increased mucus release and streaming from mucosal goblet cells with cellular cavitation compared with control loops. Sepharose-4B chromatography of amebae and cholera toxin-stimulated glycoproteins demonstrated secretion of mucins and an 80%-90% increase in low-molecular-weight proteins. E. histolytica trophozoites and cholera toxin enhanced the secretion of preformed and newly synthesized mucin glycoproteins and stimulated colonic glycoprotein synthesis. The level of mucus secretion elicited by axenic E. histolytica strains correlated with their virulence in vivo and in vitro. The amebic secretagogue was released into the culture medium and was heat stable. Mucus secretagogue activity of E. histolytica may contribute to depletion or alteration of the protective mucus blanket, facilitating pathogenesis of invasive amebiasis.

Animals↗
PubMed1991

Legal requirements for the use of phytopharmaceutical drugs in the Federal Republic of Germany.

The legal status for herbal remedies is defined by the Drug Law of 24 August 1976 in the version of the forth law amending of April 1990. For finished drugs a marketing authorization is obligatory. Herbal finished drugs hae to comply with the same criteria for quality, safety and efficacy as all other finished drugs. Marketing of finished herbal drugs is possible by three ways: 1. Procedure of evaluation and validation of old medicines. Finished drugs registered in 1978 did possess a provisional marketing authorization and could be on the market until the end of April 1990. The medical evaluation of these drugs is mainly based on bibliographic data and it is done by a special expert committee, the Commission E. It is documented by a monograph which is the basis for the assessment of these drugs starting this year. 2. Standardized marketing authorization. Medicines which do not represent a direct or indirect risk for health can be exempted from the need of an individual marketing authorization by monographs of standardized marketing authorizations. If a applicant refers to such a monograph he does not have to present any documentation. 3. Individual applications for marketing authorization. In this procedure a complete documentation including the results of analytical tests, results of the pharmacological and toxicological tests and results of clinical or other medical tests are required. Drugs sold outside pharmacies and referring only to traditional uses without clinical evidence for efficacy have to be labelled as "traditionally used".

Germany↗
PubMed1991

Autoradiographic mapping of the glucose transporter with cytochalasin B in the mammalian eye.

The anatomic localization of the glucose transport protein in the eyes of rats, rabbits, baboons, marmosets, and humans with [3H]cytochalasin B using in vitro autoradiography showed high densities of glucose carrier densities. These densities were seen in the ciliary body, especially ciliary processes, iris, retina, and in some species, the trabecular meshwork and lens. In the lens, specific [3H]cytochalasin B-binding sites were mainly concentrated in the lens nucleus. Lower concentrations were found in the cortex. During aging, glucose transporter concentration increases up to the age of 8 yr in the marmoset lens nucleus, but decreases in the cortex and retina. Moderate amounts of carrier are located in the corneal endothelium and epithelium. The enrichment of glucose carrier protein in the trabecular meshwork suggests a high metabolic activity and a possible relationship in the regulation of intraocular pressure.

Aging↗