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Biomedical subjects

R Moser

Publications and source records attributed to R Moser.

At least 37 records · Page 2Linked to original sources
PubMed1998

IL-4 and IL-13 downregulate rolling adhesion of leukocytes to IL-1 or TNF-alpha-activated endothelial cells by limiting the interval of E-selectin expression.

Leukocyte extravasation is governed by the endothelium, expressing a defined pattern of adhesion molecules in response to inflammatory stimuli. Among them, E-selectin, which is expressed in response to interleukin 1 (IL-1) or tumour necrosis factor alpha (TNF-alpha), provides rolling adhesion of the circulating leukocytes, a transient and reversible interaction that initiates leukocyte extravasation. In our experiments, E-selectin expression culminated after 4 to 6 h and declined thereafter. After 24 h a considerable amount of E-selectin was presented, reflecting its continuous expression in different inflammatory skin disorders. After preincubation of the endothelial cells with TNF-alpha together with IL-4 or IL-13, E-selectin mRNA transcription and protein expression were markedly reduced at 8 h and almost abolished at 20 h. In contrast, early E-selectin expression between 2 to 6 h was not significantly impaired. In a rotating adherence assay that mimics physiological shear forces in circulation, preincubation of the endothelial cells with TNF-alpha for 4 and 20 h induced similar adherence of neutrophils, which was largely E-selectin dependent. According to the modified expression kinetics of E-selectin in the presence of IL-4 of IL-13 rolling adhesion was unimpaired at 4 h but significantly diminished after 20 h of preincubation. Similar results were obtained with clones of the cutaneous T cell lymphoma cell line HUT78 highly expressing the E-selectin ligand cutaneous lymphocyte-associated Ag. Together, these data suggest that IL-4 and IL-13 control the rolling adhesion by limiting the period of the induced E-selectin expression and may thereby confine the acute phase of leukocyte emigration.

Antibodies, Blocking↗
PubMed1998

Soluble CD44 v5 and v6 in serum of patients with breast cancer. Correlation with expression of CD44 v5 and v6 variants in primary tumors and location of distant metastasis.

Primary breast cancers were shown to overexpress CD44 v5 and v6 at the plasma membrane. However, the clinical significance of this overexpression remains unclear. Overexpression of CD44 v5 and v6 in primary breast cancers was found to correlate with metastasis and poor prognosis by some investigators, yet this correlation could not be confirmed by others using different antibodies. In this study the influence of metastatic disease, the site of metastasis, and the amount of CD44 v5 and v6 expression in the primary tumor on serum levels of the soluble forms of CD44 v5 and v6 (sCD44 v5 and v6) in breast cancer patients was investigated. Soluble CD44 v5 and v6 serum levels were measured by enzyme linked immuno sorbent assay in a group of breast cancer patients who developed metastases in various organs and in another group of patients with single organ metastasis. For control, sCD44 v5 and v6 levels were measured in breast cancer patients who remained free of metastasis and in healthy blood donors. Expression of plasma membrane bound CD44 v5 and v6 in the primary tumors of the patients with metastasis in various organs was correlated to sCD44 v5 and v6 levels in serum. Furthermore the size of sCD44 v6 was analyzed by immunoblot using a monoclonal antibody directed against CD44 v6. When metastases were detected, sCD44 v5 and v6 serum levels were increased as compared to levels measured one month after tumor surgery in patients free of metastases (p= 0.0025 and p=0.0004). Six of 19 and 6 of 20 patients had sCD44 v5 and v6 serum levels above a cut-off level of 85 and 275 ng/mL, respectively. In these cases expression of CD44 v5 and v6 in the primary cancers was also elevated. Low sCD44 v5 and v6 serum levels were associated with weak expression of CD44 v5 and v6 in the respective primary cancers. As shown by statistical analysis of sCD44 v5 and v6 levels in 57 patients with single organ metastases, elevated sCD44 v6 levels but not sCD44 v5 levels were associated with metastases in liver or bone (p=0.0025). Immunoblot analysis of soluble CD44 proteins in serum revealed two CD44 v6 specific signals of approximately 120 and 170 kDa. Increased sCD44 v5 and v6 serum levels in patients with breast cancer were influenced by the amount of CD44 v5 and v6 expression in the primary tumor by the site of metastasis. Elevated sCD44 v6 serum levels were preferentially found in patients with metastases in liver or bone.

Adult↗
PubMed1998

Limited value of scintimammography and contrast-enhanced MRI in the evaluation of microcalcification detected by mammography.

The aim of this study was to evaluate whether scintimammography using 99Tc(m)-sestamibi or contrast-enhanced magnetic resonance imaging (MRI) can improve the specificity of mammography for the differentiation of benign and malignant breast microcalcification. From 156 consecutive patients studied with SMM, 44 patients with microcalcification on mammograms were selected for this study. Forty patients in this group also had contrast-enhanced MRI of the breast. The intensity and patterns of sestamibi uptake for scintimammography and contrast enhancement for MRI were visually determined and graded on a 5-point scale for malignancy. The results of both techniques were compared and correlated with final histopathologic diagnoses. The sensitivity and specificity of scintimammography were 63% and 85% respectively, if only those cases classified as probable or definite malignancy were considered positive. If indeterminate findings were also considered positive, the sensitivity and specificity of scintimammography were 79% and 80% respectively. Using the latter classification for MRI revealed a comparable sensitivity of 82% but a markedly lower specificity of 56%. Excluding indeterminate findings from the group of positive MRI diagnoses resulted in a specificity of 94% and a sensitivity of 64%. In conclusion, scintimammography of the breast had a comparable sensitivity but a higher specificity than MRI. The sensitivity of both techniques, however, is probably too low for routine use in the evaluation of microcalcification detected by mammography.

Adult↗
PubMed1998

[Prevalence of depressive disorders in early puerperium].

OBJECTIVE: The aims of this investigation were to detect the prevalence and influencing factors of early postpartal depressive disorders in a large hospital sample. METHODS: By means of an interview we acquired information on sociodemographic data, physical and psychiatric anamnesis, obstetric and psychological variables. The German version of the Edinburgh Postnatal Depression Scale (EPDS) served to determine the depressive disorder of our patients. The interview was carried out on 1,250 women at the maternity ward 5 days after delivery. RESULTS: According to the results of the German validation of the EPDS, 254 (20.3%) women at the maternity ward had an at least mild depressive disorder. A comparison between nondepressed women (n = 996, 79.7%) and depressed women (n = 254, 20.3%) revealed the following risk factors for the development of an early postpartal depressive disorder: higher subjective burden of childbirth, higher trait anxiety, poorer couple compatibility, lower job satisfaction and lower social status. Of all the obstetric variables previous abortions, elevated pregnancy risks, low weight of the newborn at delivery and cesarean section were of significant influence. CONCLUSIONS: Early postpartal depressive disorder appears in 20% of all women 5 days after delivery. According to previous studies this group seems to be at highest risk for developing a later postpartum depression. Considering the psychosocial and obstetric risk factors, preventive psychotherapeutic interventions at the maternity ward are indicated.

Adult↗
PubMed1998

The interleukin-4/interleukin-13 receptor of human synovial fibroblasts: overexpression of the nonsignaling interleukin-13 receptor alpha2.

Interleukin (IL)-4 and IL-13 are known to bind to shared heteromultimeric receptor complexes of variable composition. Given the many regulatory effects of IL-4 and IL-13 on synovial cells, we aimed to characterize their IL-4/IL-13 receptor (R). Cultivated synovial fibroblasts expressed transcripts for IL-4Ralpha and IL-13Ralpha1, the human homolog of the recently cloned mouse IL-13R, but not the common gamma-chain of the IL-2R. In particular, IL-13Ralpha2 mRNA, encoding a different IL-13R recently cloned from human renal carcinoma cells, was expressed at a strikingly high level. Correspondingly, a predominant protein migrating at 65 to 75 kd was cross-linked by iodinated IL-13 and was not cross-competed by an excess of unlabeled IL-4. However, by flow cytofluorometry, IL-13Ralpha1 (detected by the anti-lL-13Ralpha1 mAb 65) and IL-4Ralpha (detected by the mAb S697) were expressed at similar low density. Radioligand binding studies revealed for both cytokines approximately 300 receptors/cell with similar high affinity. An additional class of IL-13Rs was identified after occupation of the shared high-affinity receptors by the nonsignaling, double-mutant IL-4121R-->D, 124Y-->D (RY-IL-4). In these experiments, 1251-IL-13 bound to a single receptor population with a Kd of approximately 300 pM and approximately 5000 sites/cell, matching the published affinity of monomeric IL-13Ralpha2 when expressed in COS7 cells. RY-IL-4 blocked the IL-4- and IL-13-mediated vascular cell adhesion molecule (VCAM)-1 expression and Stat6 activation, suggesting that the large number of high-affinity IL-13Ralpha2 monomers are silent receptors, likely representing a decoy target for IL-13.

Carrier Proteins↗
PubMed1997

Tumor necrosis factor alpha enhances the expression of the interleukin (IL)-4 receptor alpha-chain on endothelial cells increasing IL-4 or IL-13-induced Stat6 activation.

Functional receptors for interleukin (IL)-4 and IL-13 on endothelial cells consist of the 130-kDa IL-4 receptor alpha-chain (IL-4Ralpha) and a 65-75-kDa IL-13 binding subunit that are expressed in a ratio of about 1:3, respectively. The restricted number of IL-4Ralpha limits subunit heterodimerization and in turn receptor-mediated signaling. We report here, the effects of tumor necrosis factor alpha (TNF-alpha) on the expression of the receptor subunits for IL-4 and IL-13. By flow cytofluorometry and receptor-binding analysis of iodinated IL-4 and IL-13, stimulation with TNF-alpha-induced a 2-3-fold increase of the IL-4Ralpha expression. The up-regulation was also confirmed at the transcriptional level by reverse transcription-polymerase chain reaction. Radioligand cross-linking experiments revealed no change in the subunit composition of the TNF-alpha-induced receptor complex. Nevertheless, TNF-alpha stimulation led to increased activation of the IL-4-specific signal transducers and activators of transcription protein (Stat6) by IL-4 and IL-13. Thus, TNF-alpha corrects the subunit imbalance of the endothelial IL-4.IL-13 receptor complex thereby increasing receptor heterodimerization and in turn the signaling capability by IL-4 and IL-13.

Animals↗
PubMed1997

A novel 4-kb interleukin-13 receptor alpha mRNA expressed in human B, T, and endothelial cells encoding an alternate type-II interleukin-4/interleukin-13 receptor.

A 4 kb human interleukin-13 receptor (IL-13R) chain cDNA was cloned from a B cell cDNA library using expressed sequence tags homologous to mouse IL-13R as probes. The deduced protein sequence shows a significant level of sequence identity with the IL-5R and the human IL-13R identified recently by expression cloning. The cytoplasmic region is very highly conserved between human and mouse homologs and contains a consensus binding motif for a signal transducer and activator of transcription. The cDNA encodes a protein binding IL-13 when expressed alone which participates in a receptor complex for both IL-4 and IL-13 when expressed in conjunction with the IL-4R alpha chain. Transcripts for this IL-13R chain could be detected in most tissues and organs studied and in T, B, endothelial cells, basophilic, immature mast cell, and monocytic cell lines. The pattern of expression is different from the other recently cloned IL-13R molecule, and correlates with sites where IL-4 and IL-13 signaling is known to occur. This novel receptor is therefore likely to be implicated in reactions involved in IgE responses, T helper 2 differentiation, adhesion of leukocytes to endothelium, and therefore in pathological phenomena such as allergy, atopy, and asthma.

Amino Acid Sequence↗
PubMed1997

Role of technetium-99m sestamibi scintimammography and contrast-enhanced magnetic resonance imaging for the evaluation of indeterminate mammograms.

This study evaluated and compared technetium-99m sestamibi scintimammography (SMM) and breast magnetic resonance imaging (MRI) results in patients with indeterminate mammograms to determine whether either technique can improve the sensitivity and specificity for the diagnosis of breast carcinoma. From 123 consecutive patients who underwent physical examination, mammography, SMM, and histopathologic confirmation, a subgroup of 82 patients presenting with indeterminate mammograms was studied. Sixty-eight patients underwent contrast-enhanced MRI. SMM results were scored on the basis of the intensity and pattern of sestamibi uptake. MRI images were scored on the basis of signal intensity increase after administration of contrast material as well as the enhancement pattern and speed of gadolinium uptake. The results obtained with the two techniques were compared and related to the final histopathologic diagnoses. Considering indeterminate findings as positive, the sensitivity of SMM was 79% and the specificity, 70%. MRI displayed a sensitivity of 84% and a specificity of 49%. When indeterminate results were considered negative, the sensitivity and specificity of SMM were 62% and 83%, respectively. MRI revealed a sensitivity and specificity of 56% and 79%, respectively. The calculated sensitivities and specificities demonstrate the diagnostic limitations of both SMM and MRI in the evaluation of patients with indeterminate mammographic findings. Due to the higher specificity, SMM may be the preferred modality in the evaluation of selected patients with breast abnormalities.

Breast Diseases↗
PubMed1997

Defining total corporate health and safety costs--significance and impact. Review and recommendations.

This is the first in a series of studies designed to assist directors of occupational health and safety services in defining, measuring, predicting, and integrating total health and safety costs into useful management information. This study was structured to review recent literature on health and safety costs and to categorize costs as either direct or indirect. This delineation should aid in defining total health and safety costs, delineating priority areas for interventions to reduce costs, and evaluating the effectiveness of health and safety programs. The significance of such efforts is underscored by the reported direct health care costs for the nation's work force of over $418 billion, and indirect costs, using the lower range of estimates for such costs, of over $837 billion. Reducing the total costs of over $1.256 trillion would have major impacts on corporate productivity and competitiveness, as well as on availability of health care programs for employees. Recommendations for follow-up activities to define costs and evaluate intervention programs are provided.

Costs and Cost Analysis↗
PubMed1997

Comparison of technetium-99m-sestamibi scintimammography with contrast-enhanced MRI for diagnosis of breast lesions.

UNLABELLED: Scintimammography using 99mTc-sestamibi and contrast-enhanced MRI were performed to determine the diagnostic accuracy of either method in the diagnostic workup of patients suspicious for breast tumors. METHODS: Fifty-six patients (42 with indeterminate mammograms) underwent preoperative prone planar scintimammography and pre- and postcontrast-enhanced MRI. Visually determined signal increase after application of Gd-DTPA was compared with visually scored sestamibi uptake, and the diagnoses of both methods were correlated with the final histopathologic results. RESULTS: Overall, sensitivity and specificity of scintimammography for diagnosing breast cancer were 88% and 83%, respectively. In the subgroup of patients with indeterminate mammograms, sensitivity was 79% and specificity was 83%. MRI readings provided a higher sensitivity (91% with respect to all patients and 89% with respect to patients with indeterminate mammograms), but a considerably lower specificity (52% in both groups) due to contrast-enhancement in different benign lesions. CONCLUSION: Due to its considerably higher specificity, scintimammography rather than MRI may be suitable to reduce the number of breast biopsies which yield benign results. Thus, this method may be suggested as the preferable tool in the diagnostic workup of patients with indeterminate mammographic findings.

Adult↗
PubMed1997

Evaluation of mammographic breast lesions with Tc-99m sestamibi scintimammography and contrast enhanced MRI.

Scintimammography using Tc-99m sestamibi and contrast enhanced MRI were performed in order to determine the accuracy of both methods in the diagnostic work up of patients with suspicious or indeterminate preliminary diagnosis. 25 controls and 56 patients (14 with suspicious and 42 with indeterminate preliminary diagnoses), in whom physical examination and/or mammography warranted breast biopsy, underwent prone planar scintimammography. Sestamibi uptake was scored visually and measured using the ROI technique to enable semiquantitative evaluation. The patient group additionally underwent plain and contrast enhanced MRI. Visually determined signal increase following application of Gd-DTPA was compared with scintigraphic findings and final histopathologic results. Sensitivity and specificity of semiquantitative scintimammography for diagnosing breast cancer was 88% and 87%, respectively. Based on ROC analysis a target/non target ratio R > 1.3 was shown to be the optimal threshold for separating benign from suspicious scintigraphic diagnoses. MRI reading provided a slightly higher sensitivity (91%), but a considerable lower specificity (52%) due to contrast enhancement of different benign lesions. In the clinically important patient subpopulation with indeterminate results from previous diagnostic procedures, sensitivity of scintimammography fell to 79%, while specificity remained at 87%. MRI revealed a higher sensitivity of 89% and a lower specificity of 52%. Our data indicate that semiquantitative scintimammography using Tc-99m sestamibi provides a comparable sensitivity to contrast enhanced MRI in the assessment of breast cancer. The latter does not reduce the number of biopsies yielding benign results due to the high number of false positive diagnoses. Therefore, scintimammography seems to be the preferable tool in the diagnostic work-up of patients with indeterminate mammographic diagnoses.

Aged↗
PubMed1996

Interleukin-4 (IL-4) and IL-13 bind to a shared heterodimeric complex on endothelial cells mediating vascular cell adhesion molecule-1 induction in the absence of the common gamma chain.

Interleukin-4 (IL-4) and IL-13 exert similar, nonadditive effects on endothelial cells, inducing vascular cell adhesion molecule-1 (VCAM-1) expression and subsequent transmigration of eosinophils. The receptor for IL-4 and IL-13 was described as a shared heteromultimeric complex in which the common gamma-chain (gamma c) subunit was essential for activity. Endothelial cell bound both cytokines with high affinity; by flow cytofluorometry and reverse transcription-polymerase chain reaction (RT-PCR), they expressed IL-4 receptor alpha (IL-4R alpha) but did not express the gamma c of the IL-2R. Radioligand cross-linking experiments followed by immunoprecipitation with the monoclonal antibody (MoAb) S697 to the IL-4R alpha showed IL-4-specific binding at 130 kD, the IL-4R alpha, and to a minor extent to a double band coimmunoprecipitated at 65 to 75 kD. [125 I]IL-13 bound predominantly to the 65- to 75- kD band and with a trace amount of binding at 130 kD. However, no ligand-cross-linked receptor was precipitated by the MoAb S697, indicating a cognate novel IL-13-binding subunit. Excess unlabeled IL-4 completely displaced IL-13 binding. Similarly, nonsignaling IL-4 (Y124D)-mutant abolished IL-4- and IL-13-mediated signal transduction. Unlabeled IL-13 competed successfully for IL-4 binding at 65 to 75 kD but was unable to completely displace Il-4 from its binding to the IL-4R alpha. The MoAb TUGh4, specific for the gamma c, failed to precipitate ligand-cross-linked IL-4R and IL-13R. Therefore, the subunit structure of the functional receptors for IL-4 and IL-13 on human endothelial cells does not use or require the common gamma c of the IL-2R.

Amino Acid Sequence↗
PubMed1996

The interleukin-8 receptor B and CXC chemokines can mediate transendothelial migration of human skin homing T cells.

We studied the involvement of chemokines that bind to G protein-coupled receptors in the migration of skin homing T cells across a bilayer vascular construct (BVC) consisting of a fibroblast matrix underneath an activated endothelial (EC) monolayer. Based on the expression of the cutaneous lymphocyte-associated antigen (CLA), a skin homing receptor, CD45R0+ T cells freshly isolated from blood or HUT-78 cutaneous T lymphoma cells were separated into CLA+ and CLA- subpopulations. These T cells were incubated on interleukin (IL)-1 beta and tumor necrosis factor-alpha-activated EC, and the number of transmigrated cells was determined. The chemokine IL-8 was selectively involved in the enhanced migration of CLA+ T cells across activated EC as demonstrated by blocking antibody to IL-8 but not to GRO-alpha, MCP-1 and RANTES. Identical results were obtained with both human umbilical vein EC (HUVEC) and microvascular skin EC (HDMEC). Pertussis toxin selectively inhibited the enhanced transendothelial migration (TEM) of CLA+ T cells, suggesting that CLA-dependent TEM depends on Gi protein-transmitted signals. Moreover, the IL-8 receptor B (IL-8RB) appeared to be functionally involved in TEM, as demonstrated by receptor desensitization with the CXC chemokines IL-8 and GRO-alpha and by blocking the IL-8RB with specific monoclonal antibodies. Although only the IL-8RB was involved in CLA-dependent TEM, mRNA encoding IL-8RA and IL-8RB was expressed by both CLA+ and CLA- T cells. This correlated with IL-8RA and IL-8RB surface expression on these cells. Thus, the IL-8RB is selectively functional in TEM of T cells expressing the skin homing receptor CLA. Our results demonstrate a critical role for IL-8 and possibly other IL-8RB ligands in addition to the IL-8RB in TEM and suggest the involvement of these molecules in the homing of specific T cells to inflamed skin.

Animals↗
PubMed1996

Biochemical and morphological characterization of vascular and lymphocytic interleukin-4 receptors.

The distribution of the interleukin (IL)-4 receptor in normal human and common marmoset (Callithrix jacchus) tissues was examined by immunofluorescence and flow cytometry using monoclonal antibodies specific for the human IL-4 receptor to gain further insight into IL-4-mediated inflammatory and immunological events. IL-4 receptor positivity was unequivocally demonstrated on lymphocytes, predominantly T cells, and on blood vessels in many tissues. Vascular IL-4 receptor immunofluorescence consisted of a strong smooth muscle cell positivity and weaker positive staining of capillary and venular endothelial cells. Subnanomolar concentrations of IL-4 induced a genistein-sensitive up-regulation of VCAM-1 in vascular cell cultures. Tumor necrosis factor-alpha induced a genistein-resistant up-regulation of VCAM-1. IL-4 strongly induced expression of the IL-4 receptor on splenocytes (T lymphocytes) but not on vascular smooth muscle or endothelial cell cultures. Receptor cross-linking to [125I]IL-4 revealed a 65- to 75-kDa accessory receptor subunit consistent with a recently cloned IL-13 receptor associated with the IL-4 receptor on both vascular endothelial and smooth muscle cells. The demonstration of a vascular distribution pattern for the IL-4 receptor in addition to expression on lymphocytes suggests that vascular functional alterations, transduced through a unique IL-4 receptor complex (the type II IL-4 receptor), may be of importance during immunological and allergic inflammatory events.

Animals↗
PubMed1995

Migration of skin-homing T cells across cytokine-activated human endothelial cell layers involves interaction of the cutaneous lymphocyte-associated antigen (CLA), the very late antigen-4 (VLA-4), and the lymphocyte function-associated antigen-1 (LFA-1).

The cutaneous lymphocyte-associated Ag (CLA) is expressed by a subset of circulating memory/effector T cells and by the vast majority of skin-infiltrating T cells. CLA is thought to target skin-associated T cells to inflammatory skin sites by interacting with endothelial cell ligand E-selectin (CD62E). We have examined adhesion molecules involved in the migration of human CLA+ and CLA- memory/effector T lymphocytes through IL-1- and TNF-alpha-activated and nonactivated HUVEC layers under static (nonflow) conditions. CLA-enriched memory/effector T lymphocytes migrated more actively across cytokine-activated HUVEC than CLA-depleted memory/effector T cells. This enhanced migration is dependent on the CLA/E-selectin interaction. mAb to very late Ag-4 (VLA-4) and vascular cell adhesion molecule-1 (VCAM-1) blocked the migration of CLA-enriched, but not of CLA-depleted, T cells across activated HUVEC. The observation that anti-VLA-4 and anti-CLA mAb did not show additional inhibition supports the concept that CLA and VLA-4 are sequentially involved in the extravasation. The fact that only CLA+ T cells were inhibited by the anti-VLA-4 mAb suggests that, in this system, CLA engagement is required for using the VLA-4/VCAM-1 pathway. Our studies demonstrate that CLA+ T cells use LFA-1/intercellular leukocyte adhesion molecule-1 (ICAM-1) for transmigration but that CLA expression is not required for the LFA-1/ICAM-1-dependent transmigration because anti-CD18/CD11a mAbs and anti-ICAM-1 mAbs were able to block T cell migration regardless of the activation state of HUVEC or the CLA expression by T cells. Taken together, our results suggest that CLA has a homing function in conducting the T cell to interact with LFA-1/ICAM-1 and/or VLA-4/VCAM-1; this results in enhanced adhesion and migration across cytokine-activated endothelial cells.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1995

Action of soybean lipoxygenase 1 on 12-iodo-cis-9-octadecenoic acid and 12-bromo-cis-9-octadecenoic acid.

The ferric form of soybean lipoxygenase catalyzes an elimination reaction on 12-iodo-cis-9-octadecenoic acid (12-IODE) to produce iodide ions and 9,11-octadecadienoic acid (9, 11-ODA). If excess 13(S)-hydroperoxy-cis-9,trans-11-octadecadienoic acid (13-HPOD) is present, the reaction proceeds until about one-half of the racemic 12-IODE is consumed; in the absence of excess 13-HPOD, the reaction stops after about three turnovers. Ferric lipoxygenase also catalyzes the conversion of 12-bromo-cis-9-octadecenoic acid (12-BrODE) to 9,11-ODA at a rate that is less than 25% of that observed with 12-IODE. These elimination reactions cannot be detected with ferrous lipoxygenase or with lipoxygenase that has been inactivated by 5,8,11,14-eicosatetraynoic acid. In the case of 12-IODE, elimination is accompanied by a loss of enzymatic activity; at pH 9.0, about 10 iodide ions are produced per molecule of enzyme inactivated. No inactivation can be detected with 12-BrODE. Ascorbate and hydroxylamine, which can act as free-radical traps, block the inactivation by 12-IODE but do not inhibit the elimination reaction. When the enzyme is inactivated by [1-14C]-12-IODE at pH 9.0, the amount of radioactivity that is covalently bound to the protein is less than 30% of that expected for 1:1 incorporation.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromatography, High Pressure Liquid↗
PubMed1995

Effects of lead administration on developing rat kidney. I. Glutathione S-transferase isoenzymes.

The effects of acute and chronic exposure to lead on glutathione S-transferase (GST) isoforms were determined in developing kidney in the rat. The ontogeny of glutathione S-transferase isoforms was characterized as were the effects of depletion of dietary calcium on glutathione S-transferase isoform profiles in control and lead-treated rats. In the acute exposure experiments, rats of 14 and 50 days of age received three daily injections of lead acetate (114 mg/kg) and in the chronic exposure studies, rats received lead acetate at doses ranging from 50 to 500 ppm in their drinking water. Lead acetate administration in these chronic studies began 1 day after conception. Acute and chronic lead exposure had similar effects, causing increases in all but one glutathione S-transferase isoform (Yb3); these increases were markedly exacerbated by dietary calcium depletion. In all lead paradigms, GST subunits Yb1 and Yp showed the largest increases--greater than 25-fold in rats fed a low-calcium diet. GST subunit Yb3 showed small increases in the 14-day acute lead and the 4 week low-calcium animals and did not increase in other groups. Lead-related increases in GSTs were partially reversed by transferring animals previously receiving lead to lead-free water for a 4-week period. Kidneys of rats fed the low-calcium diet did not have detectable GST Yk, but in rats on this low-calcium diet that received 500 ppm lead; this GST isoform was found at levels comparable to those in control rats fed lab chow.

Aging↗