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Biomedical subjects

A Bonhoff

Publications and source records attributed to A Bonhoff.

16 recordsLinked to original sources
PubMed1995

Identification of macroprolactin in a patient with asymptomatic hyperprolactinemia as a stable PRL-IgG complex.

We characterize the molecular form of PRL in a female patient with asymptomatic, idiopathic hyperprolactinemia. Size exclusion chromatography revealed that PRL was present exclusively in the form of macroprolactin (> 200 kD mol wt). By immunoaffinity purification, Western blot analysis, and binding studies the molecule was identified as a PRL-IgG complex of extremely high stability. Sequence analysis revealed no mutations in the protein coding region of the hPRL gene using hPRL cDNA amplified from the patient's peripheral blood lymphocytes. In spite of full bioactivity in vitro, as determined by Nb2 bioassay, the complex apparently lacks bioactivity in vivo. Its high molecular weight may reduce its access to target organs in the periphery as well as centrally.

Adult↗
PubMed1994

Modulation of prolactin secretion in human myometrium by cytokines.

Prolactin (PRL) secretion and PRL mRNA expression in human myometrial explant cultures was inhibited by the addition of human placental conditioned medium (HPCM). After 3 days treatment with 5% HPCM PRL secretion was reduced to 24% of control values (P < 0.001). The effect persisted even after removal of progesterone, which suppresses myometrial PRL production, from the HPCM. In search for additional regulatory substances present in the uteroplacental unit, we tested a number of growth factors and cytokines known to affect pituitary PRL secretion. Treatment with endothelin-3 (ET-3) at a dose of 10(-6) M was found to increase PRL release by 20% over 3 days (P < 0.05) whereas interleukin-6 (IL-6) showed no effect on PRL levels. Epidermal growth factor, vasoactive intestinal peptide and interferon-alpha (IFN-alpha) were inhibitory. Interleukin-4 (IL-4) was the most potent inhibitor of PRL expression in myometrial tissue, causing a reduction of secreted PRL to less than 50% of controls after 3 days at a dose of > or = 5 ng/ml (P < 0.001) and a concomitant reduction of PRL mRNA levels. These results demonstrate a modulation of PRL expression in the myometrium by locally present factors.

Adult↗
PubMed1993

Morphometric characteristics of endometrial biopsies after different types of ovarian stimulation for infertility treatment.

OBJECTIVE: To investigate whether various types of ovarian stimulation induce differences in endometrial development at the midluteal phase in infertile women. DESIGN: Assessment of stromal and glandular compartments in endometrial biopsies using morphometric criteria. SETTING: Institute for Hormone and Fertility Research, Hamburg, Germany. PATIENTS: The study included 18 women after treatment with human menopausal gonadotropin (hMG)/human chorionic gonadotropin (hCG) (group I), 23 women after clomiphene citrate (CC)/hMG/hCG treatment (group II), and 12 women after CC stimulation (group III). INTERVENTIONS: Endometrial biopsies and blood samples were taken simultaneously in the early to midluteal phase. To assess the time of ovulation, hormone analysis and regular checks by ultrasonography were performed. MAIN OUTCOME MEASURES: Morphometric evaluation of glandular and stromal structures revealed an impaired endometrial development after various treatment protocols. CONCLUSION: Ovarian stimulation in infertile women results in most cases in an elevation of steroid levels; however, the occurrence of an inadequate endometrial development might have an unfavorable influence on the outcome of implantation. Therefore, these findings may be of importance to the choice of treatment for infertility.

Adult↗
PubMed1992

Posttranscriptional regulation of the human prolactin gene in IM-9-P3 cells by retinoic acid.

The IM-9-P3 family of cell lines, which are derived from the B-lymphoblastoid IM-9 cell line, transcribe the human PRL (hPRL) gene by utilization of the decidual-type promoter and provide a model to study factors controlling extrapituitary expression of the hPRL gene. Here we describe regulation of hPRL gene expression in members of the IM-9-P3 family by retinoic acid (RA). When cells were incubated in medium supplemented with fetal calf serum that had been treated with dextran-coated charcoal, the addition of RA caused a 2-fold stimulation of hPRL secretion in the low hPRL-producing clone IM-9-P31 and the moderate producer IM-9-P32 (ED50, 0.53 and 0.13 nM, respectively), but not in the high hPRL-producing IM-9-P33 clone. Secretion from the RA-responsive cell lines increased steadily over the first 24 h of exposure and remained elevated for several days. The concomitant increase in hPRL mRNA steady state levels was not due to enhanced transcription of the hPRL gene, as assessed by nuclear run-on experiments, but, rather, to message stabilization. In RA-treated IM-9-P32 cells, the half-life of hPRL mRNA was significantly increased from 9 to 22 h. The transcripts were found to be preferentially associated with membrane-bound polysomes, thus being available for the secretory pathway. When we studied the expression of potential transducers of the RA signal, namely the RA receptor subtypes hRAR alpha, -beta, and -gamma and cellular RA-binding protein, we did not detect hRAR gamma or cellular RA-binding protein transcripts in the hPRL-negative clone IM-9-P6 or the hPRL-positive clones IM-9-P31, IM-9-P32, and IM-9-P33. hRAR alpha was equally expressed in all cell lines and not regulated by RA, whereas hRAR beta was differentially expressed and controlled by RA. This receptor subtype was absent from hPRL-negative members of the IM-9-P family, strongly induced by RA in the RA-responsive IM-9-P31 and IM-9-P32 cell lines via rapid transcriptional up-regulation, and only slightly induced in the RA-resistant IM-9-P33 cell line, suggesting a function in mediation of the effect of RA on hPRL gene expression.

Actins↗
PubMed1991

Decidual-type prolactin expression by the human myometrium.

The human myometrium, in addition to the decidualized endometrium of the late luteal phase and of pregnancy, has been proposed as a second source of uterine PRL, since immunoreactive PRL was found in supernatants from myometrial explant cultures. We demonstrate here that: 1) the human (h) PRL gene is expressed in the myometrium in vivo; 2) myometrial PRL is identical to pituitary hPRL; 3) the encoding transcript differs from pituitary hPRL messenger (m) RNA but is homologous to decidual and IM-9-P3 lymphoid hPRL mRNA; and 4) the expression of myometrial hPRL mRNA is inhibited by progestin. hPRL mRNA was detected in freshly isolated myometrium by Northern blot hybridization and was larger than the pituitary message. Sequence and primer extension analyses revealed that the transcript is identical to pituitary hPRL mRNA downstream of the pituitary cap site and carries an extension of the 5'-untranslated region homologous to that of decidual/IM-9-P3 lymphoid hPRL mRNA. This mRNA species results from alternative transcription initiation and comprises exon 1a of the hPRL gene, which is not transcribed in the pituitary. hPRL mRNA steady state levels and hPRL secretion increased dramatically when myometrial explants were maintained in long term culture. In addition to the 23,000 mol wt form, myometrial explants synthesized a glycosylated hPRL variant (G-hPRL) which was approximately 500 Daltons larger than pituitary G-hPRL but of similar size as lymphoid G-hPRL (26,500). Lactogenic activity of myometrial conditioned medium paralleled that of pituitary hPRL in the Nb2 lymphoma bioassay and was neutralized by the addition of monoclonal antibody to hPRL. hPRL secretion and hPRL mRNA abundance were not affected by estrogen but were markedly reduced by medroxy-progesterone acetate, which was maximally effective at a dose as low as 10(-10) M.

Adult↗
PubMed1990

Clonal derivatives of a human B-lymphoblastoid cell line producing prolactin--a cytogenetic characterization.

The IM-9-P cell line is a variant of the human B-lymphoblastoid cell line IM-9 which ectopically secretes prolactin (hPRL). The heterogeneous line IM-9-P and three sublines of clonal origin, two of them positive and one negative for PRL gene expression, were subjected to cytogenetic analysis and compared with the reference line IM-9 which showed a normal female diploid karyotype. G-banding revealed several rearrangements in the chromosomes. Nine altered chromosomes including one stable marker chromosome were common to all analysed karyotypes of IM-9-P cells and their clones. A second marker chromosome 'mar2' occurred only in the karyotypes of the hPRL producing clones, but not in the non-producing clone. None of the visible alterations involve chromosome 6 which carries the PRL gene in humans.

B-Lymphocytes↗
PubMed1990

Morphometric analysis of the endometrium of infertile patients in relation to peripheral hormone levels.

In an attempt to identify factors that may be responsible for reproductive failure, we compared endometrial biopsies taken from infertile patients during the luteal phase of spontaneous cycles (n = 18) with those taken after ovarian stimulation (n = 18). Morphometric analyses were performed and compared with peripheral estradiol (E2) and progesterone levels at the time of supposed implantation. In stimulated cycles the number of glands per square millimeter was positively correlated to the E2 level. The morphometric data point to a measurable difference between the study groups, indicating an insufficient secretory transformation of the tissue in infertile women when compared with a group of fertile women. The observations suggest that hormonal stimulation occasionally results in impaired development of the endometrial glands.

Adult↗
PubMed1986

Race:cultivar-specific induction of enzymes related to phytoalexin biosynthesis in soybean roots following infection with Phytophthora megasperma f. sp. glycinea.

Primary roots of soybean [Glycine max (L.), cv Harosoy 63] seedlings were inoculated with zoospores from either race 1 (incompatible, host resistant) or race 3 (compatible, host susceptible) of Phytophthora megasperma f. sp. glycinea (Pmg) and the activities of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), isoflavone synthase, and dihydroxypterocarpan 6a-hydroxylase related to phytoalexin (glyceollin) biosynthesis, and of glucose-6-phosphate dehydrogenase (Glc-6-PDH) and glutamate dehydrogenase (Glu-DH) were determined at various times after inoculation. About 2-4 h after inoculation with race 1, the activities of PAL, CHS, and pterocarpan 6a-hydroxylase were higher than after inoculation with race 3 and increased considerably thereafter. In contrast, activities of these enzymes in the compatible interaction were equal to or only slightly higher than in the controls over the entire infection period investigated (2-8 h). Isoflavone synthase did not increase until 7 h after inoculation with race 1. There were no significant differences in activities for Glc-6-PDH and Glu-DH between inoculated roots and controls. The results show that infection of soybean roots with zoospores of Pmg race 1 causes a race:cultivar-specific early induction of enzymes involved in glyceollin synthesis, whereas such an induction does not occur with zoospores of race 3. These findings are in agreement with the race:cultivar-specific accumulation of glyceollin in soybean roots reported previously [M. G. Hahn, A. Bonhoff, and H. Grisebach (1985) Plant Physiol. 77, 591-601].

Acyltransferases↗
PubMed1986

Chromosome analysis in polyploid human embryos.

Cytogenetic investigations have been performed on 436 unfertilized or polyploid human oocytes after in-vitro fertilization at the Department of Obstetrics and Gynaecology, University of Kiel. Thirty-two oocytes had more than two pronuclei 16-20 h after fertilization and were therefore potentially the precursors of polyploid embryos. The total number of fertilized oocytes was 667, and the frequency of tripronucleate ova was 4.8%. These tripronucleate eggs may develop normally up to birth but never lead to viable newborn children. Some of the resulting embryos displayed chromosomal mosaicism, where polyploid karyotypes and normal diploid cells occurred together. It is assumed that the survival rate of polyploid embryos depends upon the percentage of normal diploid cells.

Fertilization in Vitro↗
PubMed1986

Further investigations of race:cultivar-specific induction of enzymes related to phytoalexin biosynthesis in soybean roots following infection with Phytophthora megasperma f.sp. glycinea.

The activities of the following enzymes in soybean roots were determined at early times after infection of the roots with zoospores of an incompatible or a compatible race of Phytophthora megasperma f.sp. glycinea: dimethylallyl-diphosphate : 3,6a,9-trihydroxypterocarpan dimethylallyltransferase (prenyltransferase), an enzyme specific for glyceollin biosynthesis; NADPH-cytochrome reductase and hydroxymethylglutaryl-CoA reductase, enzymes related to the glyceollin pathway; and isocitrate dehydrogenase. Already at 4 h after infection there was a higher activity of the prenyltransferase in the incompatible interaction than in the compatible interaction, and enzyme activity in the incompatible interaction increased considerably between 4 and 8 h after infection. In the compatible interaction prenyltransferase activity was only slightly higher than in uninfected roots. The activity of the other enzymes in infected roots was not significantly different from that in the uninfected roots. No qualitative differences could be detected between the two-dimensional patterns of unlabelled proteins or proteins labelled with L-[35S]methionine of infected and uninfected roots at early times after infection. We conclude from these and earlier results (A. Bonhoff et al. (1986) Arch. Biochem. Biophys. 246, 149-154) that infection of the soybean roots with an incompatible race of the fungus leads to selective induction of the phytoalexin pathway and presumably to induction of other as yet unknown defense mechanisms.

Buffers↗
PubMed1985

Quantitative Localization of the Phytoalexin Glyceollin I in Relation to Fungal Hyphae in Soybean Roots Infected with Phytophthora megasperma f. sp. glycinea.

A radioimmunoassay specific for glyceollin I was used to quantitate this phytoalexin in roots of soybean (Glycine max [L.] Merr. cv Harosoy 63) after infection with zoospores of either race 1 (incompatible) or race 3 (compatible) of Phytophthora megasperma Drechs. f. sp. glycinea Kuan and Erwin. The sensitivity of the radioimmunoassay and an inmmunofluorescent stain for hyphae permitted quantitation of phytoalexin and localization of the fungus in alternate serial cryotome sections from the same root. The incompatible interaction was characterized by extensive fungal colonization of the root cortex which was limited to the immediate vicinity of the inoculation site. Glyceollin I was first detected in extracts of whole roots 2 hours after infection, and phytoalexin content rose rapidly thereafter. Significant concentrations of glyceollin I were present at the infection site in cross-sections (42 micrometers thick) of such roots by 5 hours, and exceeded 0.6 micromoles per milliliter (EC(90)in vitro for glyceollin I) by 8 hours after infection. Longitudinal sectioning (14 micrometers thick) showed that glyceollin I accumulated particularly in the epidermal cell layers, but also was present in the root cortex at inhibitory concentrations. No hyphae were observed in advance of detectable levels of the phytoalexin and, in most roots, glyceollin I concentrations dropped sharply at the leading edge of the infection. In contrast, the compatible interaction was characterized by extensive unchecked fungal colonization of the root stele, with lesser growth in the rest of the root. Only small amounts of glyceollin I were detected in whole root extracts during the first 14 hours after infection. Measurable amounts of glyceollin I were detected only in occasional cross-sections of such roots 11 and 14 hours after infection. The phytoalexin was present at inhibitory concentrations in the epidermal cell layers, but the inhibitory zone did not extend appreciably into the cortex. Altogether, these data support the hypothesis that the accumulation of glyceollin I is an important early response of soybean roots to infection by P. megasperma, but may not be solely responsible for inhibition of fungal growth in the resistant response.

Journal Article↗