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Biomedical subjects

M Blum

Publications and source records attributed to M Blum.

At least 199 records · Page 11Linked to original sources
PubMed1990

Effect of excess parathyroid hormone on human bone marrow fibroblasts.

Patients with uremia have excess levels of parathyroid hormone (PTH). It has been reported that excess PTH is associated with bone marrow fibrosis. The present study was undertaken to examine the in vitro effect of intact 1-34 PTH and the active 1-34 N-terminal fragment on human bone-marrow-derived fibroblasts. Proliferation of fibroblasts was not stimulated by PTH in concentrations present in uremia (5-30 U/ml) nor by uremic sera which contained either high or low PTH levels. The lack of stimulation contrasted with the activity of both PTH preparations in other systems, i.e., inhibitory to erythroid colonies and stimulatory to heart muscle cells.

Bone Marrow↗
PubMed1990

Differential regulation of type II corticosteroid receptor messenger ribonucleic acid expression in the rat anterior pituitary and hippocampus.

The present study was designed to characterize the regulation of the type II corticosteroid receptor (GR) mRNA in two tissues involved in the control of the hypothalamic-pituitary-adrenal axis. We have used a solution hybridization/S1 nuclease protection assay to quantitate GR mRNA levels in the rat hippocampus and anterior pituitary after CRF, dexamethasone (DEX), or corticosterone (CORT) treatment. In general, hippocampal GR mRNA levels increased after removal of endogenous corticosteroids by surgical adrenalectomy and decreased in response to glucocorticoid treatment. More specifically, in the hippocampus 1) GR mRNA expression was decreased when adrenalectomized (ADX) animals were replaced with a relatively low dose of CORT, but not with a low dose of DEX; 2) acutely, CRF was more effective than DEX in decreasing the levels of GR mRNA in intact animals; however, under the same paradigm in ADX animals, DEX decreased the level of GR mRNA, whereas CRF was ineffective; and 3) in contrast to the decrease in GR mRNA levels observed after acute and low doses of glucocorticoid treatment, chronic treatment with either DEX or CORT did not change the level of hippocampal GR mRNA. These results suggest that in the hippocampus the decrease in GR mRNA expression after CRF treatment is probably via the release of glucocorticoids, and that this tissue is more sensitive to endogenous glucocorticoids than DEX. Anterior pituitary GR mRNA was differentially regulated compared with that in the hippocampus. In marked contrast to Gr mRNA in the hippocampus, ADX did not alter anterior pituitary GR mRNA expression, and glucocorticoid treatment led to an increase in GR mRNA levels. In the anterior pituitary 1) glucocorticoid treatment led to an increase in GR mRNA expression, when replaced with a relatively low dose of DEX, but not when replaced with a low dose of CORT; 2) acutely, neither CRF nor DEX altered levels of GR mRNA in intact animals; however, under the same paradigm DEX increased levels in ADX animals; and 3) chronic DEX or CORT treatment of intact animals elevated levels of anterior pituitary GR mRNA. In summary, these data have demonstrated tissue-specific regulation of GR mRNA in the hippocampus and anterior pituitary, which is dependent on both the dose and length of treatment and, in addition, on the glucocorticoid itself.

Adrenalectomy↗
PubMed1990

Developmental changes in levels of proopiomelanocortin intron A-containing heterogeneous nuclear RNA and mature messenger RNA in the anterior and neurointermediate lobes of the rat pituitary.

The POMC cells of the rat pituitary undergo dynamic phenotypic changes during differentiation. Here we have determined that alterations in the relative levels of a POMC precursor RNA species and POMC mRNA occur during development and may represent another level at which the POMC phenotype is developmentally regulated. We performed solution hybridization/nuclease protection assays using a POMC exon 1/intron A splice junction probe to quantitate levels of both intron A-containing POMC heterogeneous nuclear (hnRNA) and fully processed POMC mRNA in separated anterior and neurointermediate lobes of the fetal, neonatal, and adult pituitary. The levels of POMC hnRNA per anterior lobe increased 7-fold from embryonic day 15 to adulthood (0.022 to 0.159 fmol/lobe), while POMC mRNA levels increased 121-fold (0.15 to 18.2 fmol/lobe). POMC hnRNA levels per neurointermediate lobe increased 23-fold from embryonic day 18 to adulthood (0.024 to 0.54 fmol/lobe), while POMC mRNA levels increased 69-fold (0.65 to 44.6 fmol/lobe). Thus, both anterior and neurointermediate lobes contain higher relative abundances of POMC hnRNA compared to mRNA during early development. These subsequently decrease (from 1:7 to 1: approximately 110 in the anterior lobe and from 1:27 to 1:83 in the intermediate lobe over the ages examined) as the levels of POMC mRNA in both anterior and neurointermediate lobe increase at a greater rate than POMC hnRNA as development progresses. These results provide the first measurements of POMC mRNA and hnRNA levels during ontogeny and suggest that there may be a developmental change in the regulation of POMC primary transcript processing.

Animals↗
PubMed1990

[Lowering effect of estrogen replacement treatment on immunoglobulins in menopausal women].

Similarly to the results reported in pregnancy and during oral contraceptive use, estrogen replacement therapy (ERT) had a lowering effect on serum immunoglobulins (IgG, IgA, IgM) of 20 postmenopausal women with a mean age of 52.2 years, in parallel with an increase in the blood values of estradiol (E2). The clinical implications of this phenomenon warrant the study on a bigger series and during a longer period of time. It is recommended to use with precaution the ERT in women with known defects in the immunologic system. A study on the effect of the new transdermal therapeutic system (Estraderm, TTS, Ciba) on the immunoglobulins is in course.

Adult↗
PubMed1989

Primary structure and possible origin of the non-glycosylated basic proline-rich protein of human submandibular/sublingual saliva.

Human submandibular/sublingual saliva contains one non-glycosylated basic proline-rich protein whereas parotid saliva contains multiple such components. The submandibular protein has a primary structure identical with the C-terminal segment [TZ] of the human parotid acidic proline-rich proteins that contain 150 amino acid residues (Mr 16,000). Northern-blot analyses of human parotid and submandibular glands revealed that mRNAs containing the HaeIII repeat sequence typical for acidic proline-rich proteins are expressed in both of these salivary glands whereas mRNAs for non-glycosylated basic proline-rich proteins containing a typical BstN1 repeat sequence are expressed in the parotid but not in the submandibular gland. Products of translation in vitro of mRNAs from human parotid and submandibular glands were also examined. Two immunoprecipitable bands with Mr 29,000 and 28,000 were obtained by translation of both parotid and submandibular mRNA. In the presence of microsomal membranes these proteins gave rise to proteins electrophoretically identical with the secreted acidic proline-rich proteins of Mr 16,000. These proteins were cleaved by kallikrein, giving rise to proteins with electrophoretic mobilities identical with those of a smaller acidic proline-rich protein with Mr 11,000 and peptide TZ. Additional immunoprecipitable bands with Mr ranging from 35,000 to 46,000 were seen when parotid mRNA was used for translation in vitro, and are believed to be precursors of the basic proline-rich proteins encoded by the BstN1 repeat type mRNA. Neither these bands nor a separate precursor for the basic non-glycosylated proline-rich protein was detected when submandibular mRNA was used for translation in vitro. It is suggested that the non-glycosylated basic proline-rich protein present in human submandibular saliva arises by cleavage of acidic proline-rich proteins.

Amino Acid Sequence↗
PubMed1989

A promising genomic transfectant into Xeroderma pigmentosum group A with highly amplified mouse DNA and intermediate UV resistance turns revertant.

Following transfection of genomic mouse DNA into an SV40 transformed fibroblast cell line from a patient with Xeroderma pigmentosum (complementation group A, XPA), a single UV resistant cell clone was isolated out of a total of 10(4) independent transfectants. The recipient XPA cell line has as yet not produced spontaneous revertants among 2.2 x 10(8) cells. The isolated cell clone contains 50-70 kb of mouse sequences which are heavily amplified (500-fold), and has acquired both intermediate resistance to UV killing and intermediate unscheduled DNA synthesis (UDS) capacity. By continued passage without selective pressure, cells were generated, which had lost both the dominant marker gene and repetitive mouse sequences. Single colonies of these cells were still intermediately resistant to UV suggesting that either undetected unique mouse DNA had segregated from the bulk of repetitive DNA, or, more likely, that the initially isolated transfectant was a spontaneous revertant. This documents that a persuasive clone isolated can still be a false positive (spontaneous revertant) and that an extremely laborious approach may lead into a dead end.

Animals↗
PubMed1989

Tryptophanyl-tRNA synthetase from Bacillus subtilis. Characterization and role of hydrophobicity in substrate recognition.

The tryptophanyl-tRNA synthetase from Bacillus subtilis was purified to homogeneity and characterized. It has an alpha 2 subunit structure and a molecular weight of 77,000. Tryptophanyl-tRNA synthetase does not catalyze any significant proofreading. It activates tryptophan as well as the three fluorinated analogues, DL-4-fluoro-, DL-5-fluoro-, or DL-6-fluorotryptophan (4F-, 5F-, and 6F-Trp), in the ATP-pyrophosphate exchange reaction. In the aminoacylation reaction, the fluorotryptophans act as competitive inhibitors of Trp. Their relative activities follow the same order in both reactions: Trp greater than 4F-Trp greater than 6F-Trp greater than 5F-Trp. This order is the inverse of the order of relative hydrophobicities of these compounds, pointing to the importance of hydrophobic interactions in the selective recognition by tryptophanyl-tRNA synthetase among this group of substrates. To define the physical basis of the relative hydrophobicities, the crystallographic structure of 4F-Trp was determined and compared to that of trptophan. Charge distributions calculated for tryptophan and its different fluoroanalogues on the basis of molecular structures were supported by their carbon-13 NMR spectra. Correlations between charge distributions and relative hydrophobicities suggest that the polarity of the C-F bond represents an underlying factor determining the hydrophobicities of 4F-, 5F-, and 6F-Trp, thus relating tryptophanyl-tRNA synthetase selectivity toward tryptophan and its fluoroanalogues directly to their electronic configurations.

Adenosine Monophosphate↗
PubMed1989

Primary hyperparathyroidism associated with two enlarged parathyroid glands.

An increasingly recognized although small percentage of patients with primary hyperparathyroidism have enlargement of two parathyroid glands. We have treated nine patients with primary hyperparathyroidism associated with such double parathyroid gland enlargement. In four of these patients, marked asymmetry of the two enlarged glands was noted and the failure to recognize and excise a second enlarged parathyroid gland resulted in persistent or recurrent hyperparathyroidism. In one of these patients, the second enlargement was present in a super-numerary mediastinal gland. The subsequent excision of the second enlarged parathyroid gland resulted in normocalcemia in each instance. This contrasts with five patients in whom initial excision of two enlarged glands resulted in normocalcemia with no recurrence of hypercalcemia. Only three patients fulfilled the histologic criteria of true double adenomas. The remainder showed multiglandular hypercellularity. This experience supports identifying all parathyroid glands and recognizing that even minimal enlargement of a gland may be important pathophysiologically, regardless of its histopathologic classification. Excision of both enlarged glands, even if asymmetric, is appropriate.

Adult↗
PubMed1989

Antisperm antibodies in young oral contraceptive users.

Thirty-five young oral contraceptive users (group A) and twenty-four non-users (group B) of comparable age and background were screened for the presence of serum antisperm antibodies, antibodies to Chlamydia trachomatis and vaginal infections. Twenty-three per cent (8/35) of the oral contraceptive users were found to have significant titers of antisperm antibodies as compared to none in the non-user group (p less than 0.05). In 17% of group A patients (6/35) the presence of antisperm antibodies coincided with the presence of serum antibodies to chlamydia and vaginal candidiasis. One of these patients had E. coli in her vaginal culture in addition to the serum antichlamydia antibodies. The other two patients harboring serum antisperm antibodies were free of any sign of vaginal infection. A significant correlation (r = 0.7038, p less than 0.0000005) was found between the presence of antisperm antibodies and antichlamydia antibodies. There appears to be an increased frequency of serum antisperm antibodies in oral contraceptive users. This finding coincides with infections or colonizations with potential pathogens of the lower genital tract.

Adult↗
PubMed1989

Modified Cabrol shunt for control of hemorrhage in repair of type A dissection of the aorta.

A technique of modifying the Cabrol shunt using preserved bovine pericardium and a patch of autologous pericardium to deal with postoperative hemorrhage from an ascending aortic operation is described. A fistula to the right atrium was created for autotransfusion. This simple technique is very useful for dealing with the catastrophic complication of such postoperative hemorrhage.

Anastomosis, Surgical↗
PubMed1989

Estradiol stimulates preoptic area-anterior hypothalamic proGnRH-GAP gene expression in ovariectomized rats.

The decapeptide gonadotropin-releasing hormone (GnRH) and the 56-amino acid GnRH-associated peptide (GAP) are derived from a common precursor translated from the proGnRH-GAP mRNA. Studies using solid-phase hybridization techniques (i.e., Northern blot analysis, dot blot analysis, or in situ hybridization autoradiography) have yielded a controversy as to whether estradiol stimulates, inhibits, or has any effect on proGnRH-GAP gene expression in the preoptic area-anterior hypothalamus (POA-AH) of the ovariectomized (OVX) rat. Using a sensitive and quantitative solution hybridization-nuclease protection assay, which ensures complete hybridization of target RNA to probe RNA, we examined the effects of OVX and estradiol replacement on the amount of proGnRH-GAP mRNA in individual POA-AH dissections. Rats sacrificed at different intervals after OVX showed a significant time-dependent decrease (34-60%) in the levels of POA-AH proGnRH-GAP mRNA relative to sham-operated animals; OVX rats treated with estradiol, however, had proGnRH-GAP mRNA levels comparable to those of sham-OVX animals. To verify these observations, levels of the proGnRH-GAP peptide, measured by radioimmunoassay with antibodies directed against the cleavage and amidation site between the GnRH and the GAP portions fo the precursor molecular, were also found to decrease (37%) after OVX and increase (63-85%) following estradiol replacement, relative to intact rats. These data support the view that estradiol stimulates the levels of both proGnRH-GAP mRNA and its primary translation product in the POA-AH region of the OVX rat.

Animals↗
PubMed1989

Intervening sequence-specific in situ hybridization: detection of the pro-opiomelanocortin gene primary transcript in individual neurons.

We describe here the in situ hybridization procedure which we have used to detect the pro-opiomelanocortin (POMC) gene primary transcript in nuclei of individual neurons in the periarcuate region of the hypothalamus. An exon-intron RNA probe was used to detect POMC primary transcript and mature mRNA in nuclear extracts of nucleic acids using a sensitive S1 nuclease protection assay. The levels per cell of nuclear primary transcript were similar to those seen in the anterior pituitary, suggesting that intervening sequence in situ hybridization should be feasible. A nonrepetitive complementary RNA probe specific for the first intervening sequence of the rat POMC gene (POMC IVS-A) was used to detect the POMC primary transcript in hypothalamic tissue sections by in situ hybridization. The distribution of nuclear localized autoradiographic grains was similar to that previously reported for immunocytochemically defined POMC neurons, suggesting that the procedure is also effective in brain cells.

Arcuate Nucleus of Hypothalamus↗
PubMed1989

Changes in rat pituitary nuclear and cytoplasmic pro-opiomelanocortin RNAs associated with adrenalectomy and glucocorticoid replacement.

While the transcriptional effects of glucocorticoid hormone manipulation on the pituitary pro-opiomelanocortin (POMC) gene have been documented, it is not yet clear whether glucocorticoids activate additional post-transcriptional mechanisms to regulate POMC gene expression. We have used RNA probes that span exon/intron junctions in sensitive nuclease protection assays in order to examine changes in POMC precursor RNA as well as mature mRNA in nucleus and cytoplasm following both adrenalectomy (ADX) and administration of exogenous glucocorticoids. ADX led to a rapid and sustained 8- to 10-fold increase in the level of POMC primary transcript in the anterior lobe (AL), from 1 to 14 days after ADX. Stimulation of mature POMC mRNA in the nucleus was also rapid, with 7- to 8-fold increases evident by 1 day after ADX. In sharp contrast, the time-dependent accumulation of POMC mRNA in the cytoplasm was slow in comparison, reaching levels approximately 2-fold higher than sham-operated animals by 1 day post-ADX and 12-fold higher by 14 days after ADX. Despite the constant elevated level of nuclear POMC precursor RNA, the rate of accumulation of POMC mRNA in the corticotroph cytoplasm after ADX was not linear, with the greatest increase occurring within the first 1-4 days post-ADX. This led to alterations in the molar ratio of POMC primary transcript: nuclear mRNA: cytoplasmic mRNA in the AL at 1 and 4 days after ADX and showed a relative increase in the proportion of POMC RNA transcripts within the nucleus. Acute administration of dexamethasone to ADX rats resulted in rapid 80-90% inhibition of POMC primary transcript levels in the AL that was maximal by 30 min but with no associated change in mature mRNA. No significant changes in POMC RNA were seen in neurointermediate lobe in any of these studies. These studies suggest that following ADX, time-dependent alterations in nuclear transport of mature POMC mRNA and/or changes in POMC mRNA stability, in addition to changes in gene transcription may account for the overall level of POMC mRNA expressed in the AL. Furthermore, we have illustrated the use of exon/intron probes for accurately quantitating rapid alterations in steady-state levels of nuclear precursor RNA that may reflect transcriptional responses and/or changes in post-transcriptional processing of the primary transcript.

Adrenalectomy↗
PubMed1989

Visualization of renal autoregulation in the split hydronephrotic kidney of rats.

The autoregulatory ability of the renal vascular system in the split hydronephrotic kidney was quantitated with intravital microscopy. The luminal diameters of the arcuate and interlobular arteries as well as the afferent and efferent arterioles were measured. Glomerular blood flow was determined by the dual slit technique. In the first series of experiments, the renal perfusion pressure was reduced by graded clamping of the abdominal aorta. Pressure reduction from 118 mm Hg to 95 mm Hg induced dilation of all preglomerular vessels except for the distal afferent arteriole; there was no change in the efferent arteriole and the blood flow was maintained. Further pressure reductions to 71 and 43 mm Hg caused additional dilations of the preglomerular vessels, a marginal reduction in diameter of proximal efferent arterioles and flow reductions by 15% and 41%, respectively. In the second series, systemic blood pressure was increased by continuous i.v. infusions of norepinephrine (NE). NE constricted pre- and postglomerular vessels except for the distal afferent arteriole; glomerular flow was decreased. Reduction of renal perfusion pressure during NE infusion to the preinfusion value did not diminish glomerular blood flow, but reduced the constrictor response to NE in preglomerular vessels. In a third series of experiments we examined the effect of atrial natriuretic factor (ANF) on renal autoregulation. Addition of ANF (10(-9) to 10(-7) M) to the renal bath induced a dose-dependent dilation of all preglomerular vessels and a constriction of the efferent arteriole. Pressure reduction from 120 to 95 mm Hg resulted in a further preglomerular vasodilation. These experiments demonstrate that autoregulation is mediated primarily by diameter changes in all preglomerular vessels excluding the distal segment of the afferent arteriole. Further, these data suggest that ANF induced dilation of preglomerular vessels does not impair the myogenic response of these vessels.

Acetylcholine↗