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Biomedical subjects

P Herman

Publications and source records attributed to P Herman.

At least 109 records · Page 6Linked to original sources
PubMed1995

Long-term culture of autologous transplanted bone marrow for acute myeloid leukaemia: evidence for an in vitro haemopoietic defect and lack of correlation with the speed of engraftment.

Haematological recovery after autologous bone marrow transplantation (BMT) for acute myeloid leukaemia (AML) is often delayed and available laboratory assays cannot accurately predict speed of engraftment. By using the long-term bone marrow culture (LTBMC) method, we attempted to define the haemopoietic defect underlying this slow engraftment, and assessed the usefulness of LTBMC in predicting engraftment. Cryopreserved bone marrow (BM) harvested from three different groups (AML autografts, n = 18; autografts for non-leukaemic diseases, n = 23; normal donors. n = 10) were cultured and their growth was compared and correlated with the speed of engraftment. In the AML autografts, non-adherent and adherent progenitor production was significantly reduced compared with normal BM during the whole culture period (P < 0.01). None of the LTBMC parameters was found to correlate with engraftment after autologous BMT. The non-leukaemic autografts showed progenitor production intermediate between normal and AML autografts. Their progenitor content at the end of the culture period (reflecting the stem cell pool) was not statistically different from normal BM. In this group, most of the progenitor cell contents, during LTBMC correlated with neutrophil (rs = -0.618 to -0.879, P < 0.01) and platelet (rs = -0.479 to -0.707, P < 0.02) recovery. The conclusion drawn from these results is that AML autografts are defective in their ability to sustain in vitro haemopoiesis, but this in vivo defect does not correlate with the slow haemopoietic recovery after autologous BMT. In contrast, LTBMC of autografts for non-leukaemic diseases, whose defect affects the stem cell pool to a lesser extent than BM in AML correlates with the speed of engraftment.

Acute Disease↗
PubMed1995

Nodular form of local hepatic tuberculosis: case report.

The nodular form of hepatic tuberculosis (TB) is a very uncommon disease characterized by coalescing tubercles to form tumour-like tuberculomas or abscesses without evidence of extrahepatic tuberculosis. Since 1950, 23 cases of isolated nodular TB in the liver have been reported in the world literature. In 19 of these cases laparotomy was performed and the correct diagnosis was obtained only on histologic examination. We report one additional case of multiple tuberculous abscesses which were initially diagnosed as hepatic abscesses and at intraoperative evaluation were thought to be a liver tumour. The diagnosis was made by histological examination with identification of acid-fast bacili.

Diagnosis, Differential↗
PubMed1994

Time-resolved polarized fluorescence studies of the temperature adaptation in Bacillus subtilis using DPH and TMA-DPH fluorescent probes.

The validity of the concept of homeoviscous adaptation was tested for bacteria Bacillus subtilis. The Bacillus subtilis grown at 20 degrees C (referred to as Bs20) exhibit a considerable increase of branched anteiso-C15, the major fatty acid component of membrane lipids, relative to membranes grown at 40 degrees C (Bs40). The time-resolved fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-[4-(trimethylamino)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) showed that these changes in the lipid composition are accompanied by changes in a mean lipid order. In particular, the DPH order parameters and measured in Bs20 membranes at 18 degrees C and in Bs40 membranes at 45 degrees C, respectively, tend to be equal. This effect was less pronounced for TMA-DPH. Our observations suggest that a physical parallel to the changes of lipid composition is the maintenance of an optimal lipid order in the hydrophobic core of the cytoplasmic membranes. It can be interpreted as a tendency of Bacillus subtilis to keep the lateral pressure in its membranes at an optimal value, independent of the temperature of cultivation.

Adaptation, Physiological↗
PubMed1994

Modulation of cyclic AMP production by strial marginal cells from gerbil in culture.

To further investigate the role of marginal cells (MCs) in the secretion of endolymph and because of the limitations encountered in investigating these cells in vivo, we used primary cultures of MCs derived from explants of gerbil stria vascularis and investigated modulation of the adenylate cyclase-cyclic AMP system. After 10 days on type I collagen coated plastic dishes, a confluent monolayer of epithelial-like cells was obtained which exhibited the morphologic and immunohistochemical features of the native marginal cells. The cyclic AMP (cAMP) content was determined at 37 degrees C, after 5 min of incubation with various agents, in the presence of a specific inhibitor of type III cAMP-dependent phosphodiesterase, RO 20-1724. The adenylate cyclase-cAMP system was associated with beta 2-adrenergic receptors. The cAMP content was increased by isoproterenol (23-fold), a beta-agonist, but not by octopamine, an alpha-agonist, and the affinity for ICI 118.551, a specific beta 2-antagonist, was greater than for CGP 20712A, a specific beta 1-antagonist (Kd: 0.03 x 10(-6) M and 15 x 10(-6) M respectively). The cAMP content was maximally increased by prostaglandin E2 > beta 2-adrenergic agonist >> vasopressine type 2 receptor agonist (26-, 23-, and 3-fold the basal cAMP content, respectively). The present study demonstrates that cultured marginal cells retain some of their in vivo properties including a modulated enzymatic cAMP system. This culture model should allow further in-depth investigation of the function of marginal cells.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1994

Localization of the human CD40 gene to chromosome 20, bands q12-q13.2.

CD40 is a surface glycoprotein, member of the nerve growth factor receptor family, which is expressed on B cells and plays an important role in their development, growth, and differentiation. Using chromosomal in situ hybridization, we localized the CD40 gene to the long arm of chromosome 20, bands q12-q13.2. This localization correlates well with the mapping of the murine CD40 gene to the distal region of chromosome 2, syntenic to the human 20q11-q13 region.

Antigens, CD↗
PubMed1993

Middle ear cell line that maintains vectorial electrolyte transport.

The middle ear epithelium plays a major role in keeping the temporal bone cavities fluid-free and air-filled, which is a mandatory condition to allow optimum transmission of the sound vibrations from the tympanic membrane to the inner ear. Previous works have recently established the absorptive function of the middle ear epithelium, using primary cultures derived from Mongolian gerbil (Meriones unguiculatus). Because of the paucity of cells as obtained by enzymatic digestion, we developed a middle ear cell line (MESV) using wild-type SV40 infection of primary culture of Mongolian gerbil's middle ear epithelial cells. Transformation was attested by nuclear expression of SV40 large T antigen, prolonged in vitro passages (presently beyond 50 passages), and tumor-inducing ability when subcutaneously injected in athymic mice. Transport properties were evaluated after the fifteenth passage. MESV cells retained most cardinal properties of the original middle ear epithelial cells: cell polarization was evidenced by the presence of mature junctional complexes that separate the cell membrane in two distinct domains, with apical microvilli at the luminal side, and by vectorial sodium transport responsible for the transepithelial lumen-negative potential difference (-9.3 +/- 0.14 mV in culture conditions (n = 9), -2.1 +/- 0.25 mV after overnight growth factors and serum deprivation). Short-circuit current was, like in primary cultures, mainly related to a sodium transport occurring through amiloride-sensitive apical sodium channels, since apical addition of amiloride (10(-5) M) reduced ISC from 7.0 +/- 1.4 to 0.6 +/- 0.1 microA/cm2 (P < 0.01, n = 6). Cellular cAMP content was increased by isoproterenol and prostaglandin E2 from 40.5 +/- 5.6 to 258.5 +/- 17.3 and 55.6 +/- 6.2 pmol/mg protein per 5 min, respectively (P < 0.05, n = 10). Isoproterenol and prostaglandin E2 increased ISC with very similar maximal effects: isoproterenol (10(-4) M) increased ISC from 5.73 +/- 0.31 to 12.77 +/- 0.39 microA/cm2, while prostaglandin E2 increased ISC from 5.47 +/- 0.21 to 12.87 +/- 0.42 (n = 3). Since amiloride (10(-5) M) abolished this stimulation, this may be related to an increase of the electrogenic sodium transepithelial transport. The MESV cell line could provide an interesting tool as a model of middle ear epithelial cells for the study of pathophysiological modulations of ion transport.

Amiloride↗
PubMed1993

Primary culture of Reissner's membrane from the gerbil: growth, morphologic features, and characterization.

To overcome the technical difficulties encountered in investigating the physiology of Reissner's membrane (RM), primary culture of RM was established. Minute explants obtained by mechanical dissociation were seeded on impermeable substrate coated with collagen type I in serum-free, hormone-supplemented medium. Within 2 weeks, a large cultured area was obtained which reproduced the two cell layers of the original explant. One layer, deep or superficial depending on the side to which the explant adhered, was made of cells exhibiting light and electron microscopy characteristics of an epithelium. Immunohistochemical methods revealed that they expressed cytokeratin, demonstrating their epithelial nature. The second layer was made of mesenchymal cells. This study suggests that cell culture of RM will facilitate further investigation of its physiology.

Animals↗
PubMed1993

Ion transport processes and middle ear physiopathology. An experimental approach using cell culture.

The role of the middle ear (ME) epithelium is primarily to keep the cavities air-filled and fluid-free. This condition requires elimination of mucus and cellular debris from the tympanic cavity by the ciliary activity. This clearance activity may be related to active ion transport processes in the epithelium. This paper presents a review of the experimental data suggesting the existence of ion transports in ME, and demonstrating the contribution of the culture of ME epithelial cells to the investigation of ion transport properties. Primary cultures of ME epithelial cells from Mongolian gerbils (Meriones unguiculatus) were undertaken. Polarized monolayers exhibited domes on non-porous supports. On porous supports, the absorptive function was shown to be related to a Na+ absorption from apical to basal sides of the epithelium, by means of apical amiloride-sensitive Na+ channels and basolateral Na+, K(+)-ATPase. In turn, this sodium absorption would generate an osmotic gradient and drive a water flow. An amiloride-induced chloride transport is also suggested. A hormonal modulation of these ion transport properties might play a role in the development of the mucociliary impairment that initiates chronic otitis media. The establishment of a middle ea cell line using wild-type SV 40 infection is in progress. This might provide further information on the physiology of the ME.

Animals↗
PubMed1993

[Contribution of cell culture to the study of cochlear endolymph].

Investigation of the mechanisms involved in the secretion of cochlear endolymph is hindered by the heterogeneity, the fragility, and the difficulty of access to the secretory structures of the inner ear, i.e. stria vascularis and Reissner's membrane. Cell culture may overcome these limitations and allow the study of the epithelial transport processes. Cultured cells derived from gerbil's explants of stria vascularis and Reissner's membrane presented morphological and immunohistochemical properties similar to those of the native cells. Investigation by the patch clamp technique of the apical membranes of marginal-like cells showed the presence of non specific cationic channels that could allow passive K+ flow towards endolymph and, thus, play a major role in the production of endolymph. Biochemical studies demonstrated that a cAMP production by these marginal-like cells. Primary culture of stria vascularis and Reissner's membrane is a suitable model to further investigate the physiology of endolymph production.

Animals↗
PubMed1992

Ion transport by primary cultures of Mongolian gerbil middle ear epithelium.

The transport properties of Mongolian gerbil middle ear epithelial cells grown in primary culture were studied. These cells formed polarized monolayers that exhibited domes on nonporous supports. On porous supports, monolayers developed an apical-negative transepithelial electric potential difference (VT = -37.2 +/- 2.7 mV) and a transepithelial resistance (RT = 519 +/- 56 omega.cm2). The short-circuit current equivalent (Ieq) was 62.4 +/- 6.2 microA/cm2 (mean +/- SE, n = 15). Na+ and Cl- accumulated in the basal bath and generated a basolateral hyperosmolarity that drove a net water flow. Amiloride (10 microM), when added to the apical but not to the basal bath, induced a 23.4 +/- 1.5 mV and 44.1 +/- 1.3 microA/cm2 decrease of VT and Ieq, respectively, while RT increased by 403 +/- 69 omega.cm2 (P less than 0.001, n = 15). Exposure of the monolayers to a low-Cl- solution (30 mM) enhanced the transepithelial potential, possibly by means of a Cl- secretion through apical Cl- channels. Isoproterenol (10(-4) M basolateral) increased intracellular adenosine 3',5'-cyclic monophosphate (cAMP) content (concentration of half-maximal response = 2.5 x 10(-7) M) and decreased VT, RT, and Ieq. The isoproterenol-induced fall of VT occurred even in the presence of low-Cl-solutions. This suggested an increase of the paracellular pathway conductance, although there was no significant modification of the mannitol permeability.(ABSTRACT TRUNCATED AT 250 WORDS)

Amiloride↗
PubMed1992

c-myc antisense oligonucleotides inhibit the colony-forming capacity of Colo 320 colonic carcinoma cells.

Colo 320 cells are colonic carcinoma cells known to express abundant c-myc mRNA. Based on the response of several hematopoietic cell lines to chemical inducers of differentiation, we reasoned that such agents might have similar inductive activity in Colo 320 cells. Accordingly, we exposed Colo 320 cells to 5 mM sodium butyrate (NaBT) for 7 d. C-myc expression decreased threefold and self-replicative potential decreased (defined as a greater than 60% decrease in colony-forming capacity in soft agar that did not contain inducer). In an effort to demonstrate a direct cause and effect between myc expression and the colony-forming capacity of Colo 320 cells, we exposed these cells to a 15-base antisense c-myc oligonucleotide (complementary to the translation initiation region of exon II). Cells were also exposed to equimolar (20 microM) amounts of sense and missense oligonucleotides. Subsequently, cells were incubated at 10, 20, 30, and 40 microM antisense DNA for 16 h, then washed and plated in oligonucleotide-free agar medium. We demonstrated that: (a) the oligomers were stable in the extracellular medium and in the cell cytoplasm; (b) the uptake of the oligonucleotides was 0.7%; (c) sense and missense oligonucleotides had no effect on colony-forming capacity; and (d) the antisense c-myc oligonucleotide resulted in a 40-75% concentration-dependent decrease in colony-forming capacity. The specific inhibition of colony-forming capacity by antisense DNA suggests that the role of myc expression in Colo 320 cells is similar to its role in hematopoiesis, and that the failure to inhibit myc expression maintains colony-forming capacity. This system provides a new strategy for inducing differentiation and may provide further insight into the genetic factors that govern the process of colonic carcinogenesis.

Butyrates↗
PubMed1991

A new method of measuring dimensional change.

A new method of measuring linear dimensional change in denture base resins is presented and its precision in determining distances between two points was investigated. Chrome steel ball bearings 1.57 mm in diameter were invested in a stone mold and were used to record measurements by a reflection from the surface of the ball bearings. The accuracy of this method was determined by performing repeated measurements between two points in a stone mold. Random measurements were made to an accuracy of 0.002 mm. The dimensional change of light-cured, heat-cured, and self-cured denture base resins was also determined with this method, followed by an analysis of variance of the data. The findings support the accuracy and reproducibility of this new method in recording linear measurements. The dimensional changes of all three resins were different from each other after the specimens were processed and stored in distilled water for 7 days at 37 degrees C.

Acrylic Resins↗
PubMed1991

A model for cell-type determination and differentiation in plants.

We are using trichome formation on the plant Arabidopsis thaliana as a model for the study of plant cell determination and differentiation. Several of the genes that are required for trichome formation are defined by mutations. Two mutations, ttg and gl1, prevent the initiation of trichome differentiation. Thus, these mutations define products that are involved in the signalling of trichome determination. Other mutations, gl2, gl3, dis1, and dis2, define genes that are involved in trichome maturation. Our immediate goal has been to isolate the genes defined by these mutations and determine the role that they play in trichome formation. Our general goals are (1) to identify counterparts to these genes that are involved in other cell type determination and differentiation processes; (2) to manipulate cell development by altering the normal expression of these genes; and (3) to determine if this information can be used to improve crop plants. Presently, most of our progress has centered on the GL1 gene, which has been isolated and characterized. We have found that GL1 is a myb-related gene that is uniquely required for trichome initiation. As in other plants, Arabidopsis has a family of myb-related genes. We are currently investigating the possibility that some of these other myb-related genes are also uniquely required for other types of cell determination events.

Arabidopsis↗
PubMed1991

[Culture of the epithelium of middle ear in the study of seromucous otitis].

Preliminary results on transport properties of middle ear epithelium in primary culture are reported. Primary cultures of Mongolian gerbil (Meriones unguiculatus) middle ear epithelial cells were undergone. These cells formed polarized monolayers that exhibited domes when grown to confluence on non porous support. Domes are the consequence of active transepithelial transports of solutes and water. On porous support, Na+ and Cl- accumulated in the basal bath and generated a basolateral hyperosmolarity that drove a net water flow. This solutes and water absorption was the consequence of an active Na+ transcellular transport, by means of apical sodium channels, that was responsible for the transepithelial potential difference, lumen negative: -39 +/- 1.2 mV (mean +/- SEM, n = 63). Transport of fluid and ions in the middle ear cavities could play a major role in keeping the cavities fluid-free and air-filled and modulating the mucociliary clearance. Modulation of these transports could be involved at the start of otitis media with effusion, and would contribute to the mucociliary clearance impairment.

Absorption↗