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E Friedman

Publications and source records attributed to E Friedman.

At least 217 records · Page 12Linked to original sources
PubMed1995

Colon absorptive epithelial cells lose proliferative response to TGF alpha as they differentiate.

As colon epithelial cells migrate up the cylindrical colonic crypt, they terminally differentiate and lose their ability to divide. Elevated levels of the epithelial cell mitogen TGF alpha have been found at the top of the crypt by other investigators, causing us to speculate that colon epithelial cells lose mitogenic response to TGF alpha as they differentiate. We tested this hypothesis by using the HT29 colon carcinoma sublines U4 and U4H as models of one colonocyte lineage, fluid-transporting enterocytes. TGF alpha was mitogenic for the U4 cells, but inhibited the growth of the more differentiated U4H cells. However, p44 MAP kinase was activated by TGF alpha in both U4 and U4H cells, as well as in two control undifferentiated HT29 sublines which showed no change in proliferation in response to TGF alpha. In addition, TGF alpha activated the EGF receptor in each line by increasing its tyrosine phosphorylation. No relationship was found in these four lines between response to TGF alpha and level of expression of either the EGF receptor or two EGF receptor ligands, TGF alpha and amphiregulin. Activated EGF receptors initiate both growth-inhibitory and mitogenic signals in these cells since blocking some of the EGF receptors on TGF alpha-growth-inhibited U4H cells and TGF alpha-unresponsive U9 cells overrode the inhibitory signals and made both U9 and U4H cells sensitive to mitogenesis by added TGF alpha. These data imply that upon reaching stages of greater maturation, colon enterocytes lose proliferative response to TGF alpha because of changes in signaling by their EGF receptors.

Amphiregulin↗
PubMed1995

Ocular blood flow velocity in age-related macular degeneration.

BACKGROUND: Changes in the structure of the ocular blood vessels associated with age-related macular degeneration (AMD) have been described in some detail, but comparatively little is known of the concomitant circulatory changes. The goal of this study is to evaluate changes in the ocular circulation that may be associated with AMD. METHODS: Ocular blood flow velocities and vessel pulsatilities were measured in volunteers with and without AMD using a color Doppler imaging unit. Spectral analyses were recorded from the ophthalmic artery, central retinal artery and vein, the temporal and nasal short posterior ciliary arteries, and the four vortex veins. RESULTS: Adjusting for age, pulsatility indices of all arteries were higher in subjects with AMD (central retinal artery [P = 0.02]; temporal and nasal short posterior ciliary arteries [P = 0.06 and 0.002, respectively]; and ophthalmic artery [P = 0.24]). End-diastolic blood flow velocity of the short posterior ciliary arteries tended to decrease in the presence of AMD. CONCLUSIONS: The combination of increased pulsatility and decreased velocity of the short posterior ciliary arteries, observed in the presence of AMD, are interpreted as evidence of increased vascular resistance. The clinical signs of AMD may be related to degradation of the metabolic transport function of the retinal pigment epithelium, resulting from impaired choroidal perfusion.

Aged↗
PubMed1995

Dopamine receptor stimulation decreases cytosolic gamma protein kinase C immunoreactivity in rat hippocampal slices: evidence for increased Ca(2+)-dependent proteolysis.

The effect of dopamine (DA) receptor stimulation on the distribution of gamma protein kinase C (gamma PKC) in hippocampal slices was assessed. Nanomolar concentrations of DA decreased cytosolic gamma PKC (56%) without altering membrane gamma PKC levels, resulting in decreased total gamma PKC immunoreactivity. The maximal decrease in cytosolic gamma PKC occurred at 20 min of incubation and was significantly blocked by the D1 DA antagonist SCH 23390 (10(-6) M) but not by the D2 antagonist sulpiride (10(-5) M). The D1 agonists SKF 38393 and A 77636 mimicked the effect of DA with similar responses produced at 10 microM and 1 nM, respectively. The D2 agonist quinpirole had no effect on gamma PKC immunoreactivity, thus indicating that this dopaminergic response is mediated through a D1-like receptor. DA had no effect on alpha, delta, or zeta PKC isozyme immunoreactivity in the same hippocampal preparations. The DA-induced decrease in cytosolic gamma PKC immunoreactivity was blocked by the Ca(2+)-dependent protease inhibitor N-acetyl-Leu-Leu-norleucinal (100 microM) and by the inorganic Ca2+ channel blocker Co2+. The data suggest that DA stimulates a D1-like DA receptor, which increases the influx of Ca2+ and activates the Ca(2+)-dependent proteolysis of gamma PKC.

Animals↗
PubMed1995

Colon carcinoma cells with inactive nm23 show increased motility and response to motility factors.

nm23H1 has properties of a metastasis suppressor gene. Although its mechanism of action is unknown, nm23 has been implicated in transforming growth factor beta 1 (TGF beta 1) signal transduction. In an earlier study we decreased nm23 mRNA levels 2- to 8-fold by antisense phosphorothiolated oligonucleotides in two HT29 colon carcinoma sublines at different stages in tumor progression with different responses to TGF beta 1: the HD3 subline, which shows TGF beta 1-induced growth arrest and differentiation; and the more tumorigenic U9 subline, whose growth and invasion are stimulated by TGF beta 1. Only TGF beta 1-mediated responses in HD3 cells were inhibited by nm23 antisense oligos, suggesting that nm23 functions in only one TGF beta 1 signaling pathway. In the current report we have extended this study to cell motility. HD3 motility was increased by nm23 phosphorothiolated antisense oligos which decrease nm23 mRNA levels, while HD3 cell motility was conversely decreased by TGF beta 1 which increases nm23 mRNA levels. HD3 motility was not increased by basic FGF, TGF beta 1 or TGF alpha, while the 13-fold higher basal motility of U9 cells was stimulated 3-fold by basic FGF, 4-fold by TGF beta 1 and 5-fold by TGF alpha, but not by scatter factor. Differences in motility and response to motility factors could not be ascribed to differences in either basal levels of proteases or modulation of their levels by TGF beta 1. Both HD3 and U9 cells displayed equal levels of urokinase activity and mRNA, equal expression of the metalloproteinase inhibitor TIMP-1, and no detectable collagenases by zymography. No differential response to TGF beta 1 was seen in any of these assays. Thus limited cell motility and lack of response to motility factors in HD3 colon cancer cells could be correlated with expression of nm23 active in signal transduction.

Base Sequence↗
PubMed1995

Expression of G protein alpha subunits in the aging cardiovascular system.

Alterations in responsiveness to stimulation of G protein-coupled receptors in the cardiovascular system have been noted during aging and may be due to changes in the G proteins. The levels of G protein alpha subunits and their mRNAs were examined in the aorta and heart of adult (6-month-old) and senescent (24-month-old) Fischer 344 rats. Western blot analysis using anti-Gs alpha antiserum revealed 4 bands in the aorta (38, 42, 45, and 52 kD). The 42 kD band decreased significantly in 24-month-old rats, whereas the other bands did not change. Western blots using anti-Go alpha and anti-Gi alpha revealed single bands of 39 kD and 41 kD respectively, which decreased significantly in the aortas from 24-month-old rats. There were no significant changes in the levels of G alpha mRNAs in the aorta during aging. In the heart, anti-Gs alpha antiserum detected 3 bands (42, 45, and 52 kD) which did not change with age. Levels of Gi alpha (41 kD) and Go alpha (39 kD) also did not change with age in the heart. The mRNAs for Gi2 alpha and Gi3 alpha were significantly increased in the heart, but there was no change in Go alpha or Gs alpha mRNAs. Reduced expression of Gi alpha may provide the molecular basis for alterations in receptor responsiveness in the aging vasculature. Altered expression of G protein alpha subunits does not appear to account for age-related alterations in cardiac function.

Aging↗
PubMed1995

Exercise reduces age-dependent decrease in platelet protein kinase C activity and translocation.

BACKGROUND: This study examined the effect of age and aerobic exercise performed on a regular basis on human platelet protein kinase C (PKC) activity and translocation. METHODS: Blood platelets were obtained from young (20-36 years) and older (61-78 years) healthy male human subjects. Platelets were incubated with PMA, 5-HT, or thrombin; PKC activities were measured in partially purified extracts of cytosolic and membranous fractions. RESULTS: Platelet PKC activities associated with both the membranes and the cytosol cellular fractions were significantly reduced in older nonexercisers. Redistribution of platelet PKC activity elicited by stimulating the cell surface receptors for 5-HT or thrombin or by direct PKC stimulation (PMA) was found to be reduced in the elderly subjects. Maintenance of aerobic fitness in the older group of exercisers partially prevented the age-associated decline in platelet PKC activity and in stimuli-induced enzyme redistribution. CONCLUSIONS: These results indicate that platelet PKC activity and its translocation may be biological markers of aging and that aerobic exercise may serve to slow the rate at which enzyme activation declines during senescence.

Adult↗
PubMed1995

Effects of dietary restriction on the change in aortic alpha 1-adrenoceptor mediated responses during aging in Fischer 344 rats.

These studies examine changes in alpha 1-adrenoceptor-stimulated contraction, accumulation of inositol phosphates (IPs) and calcium influx during aging, and the effect of dietary restriction. The potency of norepinephrine (NE) at stimulating aortic contraction was highest in aortas from 1-month-old rats compared to 6- or 24-month-old rats, while the potency at stimulating IP accumulation was higher in 6- and 24-month-old rats. The fact that the NE potency for IP accumulation is not decreased with age and is even increased a little, indicates that PI hydrolysis is not limiting for contraction. The data from 24-month-old dietary restricted rats support the same idea. Dietary restriction greatly increased the potency of NE for IP accumulation in the old animals (by 20-fold), but did not restore potency for contraction. Nifedipine (1 microM), a calcium channel blocker, inhibited the NE-stimulated aortic contractile response by 28% in 1-month, 40% in 6-month, and 67% in 24-month-old rats. While nifedipine did not inhibit NE-stimulated IP accumulation in 1-month-old aortas, it inhibited by 30% in 6-month-old aortas and by 27% in 24-month-old aortas. Dietary restriction (DR) did influence the inhibitory effects of nifedipine. Nifedipine inhibition of NE-stimulated contraction in 24-month-old DR rats was comparable to the inhibition in 6-month-old ad libitum (ad lib) controls and was less than in 24-month-old controls. Furthermore, nifedipine was less effective at inhibiting NE-stimulated IP accumulation in aortas from 24-month-old DR rats compared to 24-month-old controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1995

Candidate genes for multiple endocrine neoplasia type 1.

The aim of this study was to isolate and characterize candidates for the multiple endocrine neoplasia type 1 (MEN1) gene. The development of tumours related to MEN1 is associated with somatic deletions involving the MEN1 locus, suggesting inactivation of a tumour-suppressor gene in this region. We have isolated five cDNA candidates located within the 900 kb remaining for the MEN1 gene, determined their sequence, and characterized their expression in normal tissues and several endocrine tumours. One of the candidates, encoding for phospholipase C-beta 3, showed properties consistent with the idea of a tumour-suppressor gene.

Blotting, Northern↗
PubMed1995

The role of ras GTPase activating protein in human tumorigenesis.

Abnormal signal transduction involving activated ras genes plays a major role in the development of a variety of tumors. Ras GTPase-activating protein (rasGAP) is a major contributor to the downregulation of ras by facilitating GTP hydrolysis of activated ras. In addition, GAP participates in the down-stream effector system of the ras signaling pathway. Thus, depending on the precise genetic alteration, its location within the gene and the effects it exerts on protein function, rasGAP can theoretically function as either an oncogene or as a tumor suppressor gene. The putative role that rasGAP plays in human tumorigenesis is further emphasized by two lines of indirect evidence. First, mutations within the C-terminal SH2 region of rasGAP in a subset of basal cell carcinomas were demonstrated. These are presumably activating mutations and therefore confer a direct oncogenic potential to rasGAP. Second, an inverse correlation between rasGAP protein expression and the invasive/malignant potential in human trophoblastic tumors was shown. Thus, in these latter tumors rasGAP functions as an apparent tumor suppressor gene. Employing combined laboratory techniques and approaches to a variety of human tumors will further define the role of rasGAP in tumorigenesis, provide insight into the mode of action of rasGAP and structure-function relationships. Furthermore, it will help in establishing genotype-phenotype correlations and potentially may lead to a pharmacological approach to treating choriocarcinomas.

Basal Cell Carcinoma↗
PubMed1995

Lack of enhanced T-helper cell function in uremic patients with circulating alloreactive antibodies.

Some uremic patients with a history of blood transfusion, pregnancy, and previous transplantation maintain high levels of alloreactive cytotoxic antibodies in the absence of continuous exogenous allogenic stimuli and are thus considered sensitized to the major histocompatibility proteins. To differentiate into antibody-producing cells, B lymphocytes must interact with T-helper (CD4+) cells. Whether ongoing help from these cells is necessary for the B cells to continue producing cytotoxic alloreactive antibodies in these sensitized uremic patients is unknown. To gain insight into the cellular mechanisms that are associated with sustained alloantibody production, T cell activation markers were measured and specific and nonspecific T-helper cell function was studied in three uremic groups with different levels of panel reactive antibodies: 10 patients whose sera reacted to more than 80% of a panel of normal lymphocytes for at least 6 months before the study were highly sensitized, 20 patients whose sera reacted to less than 80% of the panel were moderately sensitized, and 10 nonsensitized patients whose sera did not react to any cell on the panel. The number of total and activated T-helper cells was similar in the highly sensitized and nonsensitized patients. Peripheral blood lymphocyte proliferation in response to plant lectins, soluble OKT3, or alloantigens was similar in the three uremic groups. The spontaneous proliferation of pure T-helper cells and proliferative responses to immobilized OKT3 or alloantigens were also similar in highly sensitized and nonsensitized patients. Alloreactive interleukin-2-producing cell frequencies with pure CD4+ cells as responding cells were 771 +/- 77.9/10(6) cells in highly sensitized, 945 +/- 252/10(6) cells in nonsensitized, and 973 +/- 114/10(6) cells in controls (P = not significant). Panel reactive antibody levels did not correlate with any of the measures of T helper responses. There was a significant decrease of peripheral blood lymphocyte responses to alloantigens and anti-CD3 antibody in all uremic patients as compared with normals, suggesting a dysfunction in accessory cells that was quantitatively similar in sensitized and nonsensitized patients. In spite of the continuous production of alloantibodies by B cells, there is no evidence of either specific or nonspecific enhancement of T-helper cell function in sensitized patients. The absence of T cell immunity to alloantigens suggests that sustained activation of T-helper cells with subsequent interleukin-2 production is not necessary to maintain alloreactive B cell function.

Adult↗
PubMed1995

The threat of time.

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Health Care Reform↗
PubMed1995

Prenatal exposure to cocaine selectively reduces D1 dopamine receptor-mediated activation of striatal Gs proteins.

The effect of in utero exposure to cocaine on striatal dopamine receptors was assessed at postnatal days 10 through 100 by examining receptor-mediated increases in GTP binding to G alpha proteins. Pregnant Dutch-belted rabbits were injected with 4 mg/kg i.v. of cocaine HCl twice a day on gestational days 8 through 29, and striatal membranes were prepared from their progenies on days 10 through 100. Dopamine-stimulated [35S]GTP gamma S binding to membrane alpha subunits was measured and found to increase binding to G alpha s and G alpha i. Pharmacological characterization of the dopamine response revealed that enhanced [35S]GTP gamma S binding to G alpha s is associated with D1 receptor stimulation, whereas binding to G alpha i is linked to D2 receptor activation. The abilities of dopamine to stimulate the binding of [35S]GTP gamma S to G alpha s but not to G alpha i was reduced in striata obtained from cocaine-exposed animals when examined at 10, 50 or 100 days of age. Similarly, prenatal cocaine exposure also reduced dopamine-stimulated [alpha-32P]GTP binding to G alpha s without influencing binding to G alpha i. Fetal cocaine exposure did not change carbachol-induced increases in [35S]GTP gamma S binding to G alpha i and G alpha o. Immunoblot analyses showed no changes in the amounts of these alpha subunits in membranes from cocaine-exposed animals vs. controls. Moreover, prenatal cocaine did not affect [3H]SCH23390 binding to D1 dopamine receptors in the caudate, putamen or substantia nigra.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗
PubMed1995

Beta-adrenoceptor-G alpha S coupling decreases with age in rat aorta.

beta-Adrenoceptor (beta AR) responsiveness, receptor density, receptor-G protein coupling, and the possible role of membrane fluidity in receptor-G protein coupling were investigated in the rat aorta with age. The beta AR agonist isoproterenol (ISO) produced relaxation of KCl-induced aortic contractions by 97%, 21%, and 0% in aortae from 1- 6-, and 24-month-old Fischer 344 rats, respectively. Forskolin completely relaxed the contractions at all ages. beta AR density was determined in aortic membranes by saturation binding of 125I-cyanopindolol (125I-CYP). beta AR density was 76, 52, and 47 fmol/mg of protein in 1-, 6-, and 24-month-old rats, respectively. To investigate beta AR coupling to G proteins, displacement by ISO of 125I-CYP binding was determined in aortic membranes in the presence and absence of the GTP analog guanosine-5'-(beta gamma-imido)triphosphate [Gpp(NH)p] (0.1 mM). The effect of Gpp(NH)p on the ISO displacement curve for 125I-CYP binding was greatest in 1-month-old rats and decreased markedly with age. In 1-month-old aorta, in the absence of Gpp(NH)p the ISO displacement curve was biphasic and two affinity constants were determined (KH - 0.061 microM and KL = 2.4 microM). In the presence of Gpp(NH)p the ISO displacement curve was monophasic (Kd - 0.72 microM). In 6-month-old aorta, whereas an effect of Gpp(NH)p on the ISO displacement curve could still be observed [in the absence of Gpp(NH)p, KH = 0.2 microM and KL = 3.5 microM; in the presence of Gpp(NH)p, Kd - 0.83 microM], the affinity constant for high affinity agonist binding and the percentage of receptors with high affinity for agonist were decreased significantly. In 24-month-old aorta there was no effect of Gpp(NH)p on the ISO displacement curve and a single affinity constant was detected [0.7 microM and 0.8 microM in the presence and absence of Gpp(NH)p, respectively]. The presence of two affinity constants for ISO in 1- and 6-month-old aorta in the absence of Gpp(NH)p and single affinity constants in the presence of Gpp(NH)p presumably represent the G protein-coupled and uncoupled states of the beta ARs, which are not observed in 24-month-old aorta. The ability of the beta AR to form the high affinity nucleotide-sensitive complex with the agonist was restored by treatment of the membranes with cis-vaccenic acid, which increases the fluidity of the membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗