4-Hydroxylation of estradiol by human uterine myometrium and myoma microsomes: implications for the mechanism of uterine tumorigenesis.

Estradiol is converted to catechol estrogens via 2- and 4-hydroxylation by cytochrome P450 enzymes. 4-Hydroxyestradiol elicits biological activities distinct from estradiol, most notably an oxidant stress response induced by free radicals generated by metabolic redox cycling reactions. In this study, we have examined 2- and 4-hydroxylation of estradiol by microsomes of human uterine myometrium and of associated myomata. In all eight cases studied, estradiol 4-hydroxylation by myoma has been substantially elevated relative to surrounding myometrial tissue (minimum, 2-fold; mean, 5-fold). Estradiol 2-hydroxylation in myomata occurs at much lower rates than 4-hydroxylation (ratio of 4-hydroxyestradiol/2-hydroxyestradiol, 7.9 +/- 1.4) and does not significantly differ from rates in surrounding myometrial tissue. Rates of myometrial 2-hydroxylation of estradiol were also not significantly different from values in patients without myomata. We have used various inhibitors to establish that 4-hydroxylation is catalyzed by a completely different cytochrome P450 than 2-hydroxylation. In myoma, alpha-naphthoflavone and a set of ethynyl polycyclic hydrocarbon inhibitors (5 microM) each inhibited 4-hydroxylation more efficiently (up to 90%) than 2-hydroxylation (up to 40%), indicating > 10-fold differences in Ki (<0.5 microM vs. > 5 microM). These activities were clearly distinguished from the selective 2-hydroxylation of estradiol in placenta by aromatase reported previously (low Km, inhibition by Fadrozole hydrochloride or ICI D1033). 4-Hydroxylation was also selectively inhibited relative to 2-hydroxylation by antibodies raised against cytochrome P450 IB1 (rat) (53 vs. 17%). These data indicate that specific 4-hydroxylation of estradiol in human uterine tissues is catalyzed by a form(s) of cytochrome P450 related to P450 IB1, which contribute(s) little to 2-hydroxylation. This enzyme(s) is therefore a marker for uterine myomata and may play a role in the etiology of the tumor.

Identification of receptor-binding domains on human interleukin 5 and design of an interleukin 5-derived receptor antagonist.

A detailed structure-function analysis of human interleukin 5 (hIL5) has been performed. The hIL5 receptor is composed of two different polypeptide chains, the alpha and beta subunits. The alpha subunit alone is sufficient for ligand binding, but association with the beta subunit leads to a 2- to 3-fold increase in binding affinity. The beta chain is shared with the receptors for IL3 and granulocyte/macrophage-colony-stimulating factor--hence the descriptor beta C (C for common). All hIL5 mutants were analyzed in a solid-phase binding assay for hIL5R alpha interaction and in a proliferation assay using IL5-dependent cell lines for receptor-complex activation. Most residues affecting binding to the receptor alpha subunit were clustered in a loop connecting beta-strand 1 and helix B (mutants H38A, K39A, and H41A), in beta-strand 2 (E89A and R91A; weaker effect for E90A) and close to the C terminus (T109A, E110A, W111S, and I112A). Mutations at one position, E13 (Glu13), caused a reduced activation of the hIL5 receptor complex. In the case of E13Q, only 0.05% bioactivity was detected on a hIL5-responsive subclone of the mouse promyelocytic cell line FDC-P1. Moreover, on hIL5-responsive TF1 cells, the same mutant was completely inactive and proved to have antagonistic properties. Interactions of this mutant with both receptor subunits were nevertheless indistinguishable from those of nonmutated hIL5 by crosslinking and Scatchard plot analysis of transfected COS-1 cells.

[Forensic dispute on thinking and the soul], undated

One folio-sized leaf containing a handwritten essay responding to an unidentified opponent's claims that "thinking is essential to the soul." The response begins with the introduction, "In the consideration of this question, I shall only examine one or two of the most material objects of our antagonist." The verso is inscribed: "2d Forensic. not read."

Human creation, imagination and autonomy: a brief introduction to Castoriadis’ social and psychoanalytical philosophy

During the last decade Castoriadis’ questioning has become a reference point in contemporary social theory. In this article I examine some of the key notions in Castoriadis’ work and explore how he strives to develop a theory on the irreducible creativity in the radical imagination of the individual and in the institution of the social-historical sphere. Firstly, I briefly discuss his conception of modern capitalism as bureaucratic capitalism, a view initiated by his criticism of the USSR regime. The following break up with Marxist theory and his psychoanalytic interests empowered him to criticize Lacan and read Freud in an imaginative, though unorthodox, fashion. I argue that this critical enterprise assisted greatly Castoriadis in his conception of the radical imaginary and in his unveiling of the political aspects of psychoanalysis. On the issue of the radical imaginary and its methodological repercussions, I’m focusing mainly on the radical imagination of the subject and its importance in the transition from the ‘‘psychic’’ to the ‘‘subject’’. Taking up the notion of “Being” as a starting point, I examine the notion of autonomy, seeking its roots in the ancient Greek world. By looking at notions such as “praxis”, “doing”, “project” and “elucidation”, I show how Castoriadis sought to redefine revolution as a means for social and individual autonomy. Finally I attempt to clarify the meaning of “democracy” and “democratic society” in the context of the social imaginary and its creations, the social imaginary significations.

A Rapid FACS-Based Strategy to Isolate Human Gene Knockin and Knockout Clones

Gene targeting protocols for mammalian cells remain inefficient and labor intensive. Here we describe FASTarget, a rapid, fluorescent cell sorting based strategy to isolate rare gene targeting events in human somatic cells. A fluorescent protein is used as a means for direct selection of targeted clones obviating the need for selection and outgrowth of drug resistant clones. Importantly, the use of a promoter-less, ATG-less construct greatly facilitates the recovery of correctly targeted cells. Using this method we report successful gene targeting in up to 94% of recovered human somatic cell clones. We create functional EYFP-tagged knockin clones in both transformed and non-transformed human somatic cell lines providing a valuable tool for mammalian cell biology. We further demonstrate the use of this technology to create gene knockouts. Using this generally applicable strategy we can recover gene targeted clones within approximately one month from DNA construct delivery to obtaining targeted monoclonal cell lines.

Federal aid : programs available to state and local governments : fact sheet for the chairman, Subcommittee on Labor, Health and Human Services, Education, and Related Agencies, Committee on Appropriations, U.S. Senate /

"B-243721"--P. 1.

Guidelines for prevention of transmission of human immunodeficiency virus and hepatitis B virus to health-care and public-safety workers : a response to P.L. 100-607, The Health Omnibus Programs Extension Act of 1988.

Item 499-F-3.

Departments of Labor, Health and Human Services, Education, and related agencies appropriations for 2012 : hearings before a subcommittee of the Committee on Appropriations, House of Representatives, One Hundred Twelfth Congress, first session /

Shipping list no.: 2011-0418-P (pt. 1), 2011-0423-P (pt. 2A), 2011-0426-P (pt. 2B), 2011-0439-P (pt. 3), 2011-0432-P (pt. 4), 2012-0085-P (pt. 5), 2012-0221-P (pt. 6), 2012-0256-P (pt. 7).

Departments of Labor, Health and Human Services, Education, and related agencies appropriations for 2011 : hearings before a subcommittee of the Committee on Appropriations, House of Representatives, One Hundred Eleventh Congress, second session /

Mode of access: Internet.

Departments of Labor, Health and Human Services, Education, and related agencies appropriations for 2013 : hearings before a subcommittee of the Committee on Appropriations, House of Representatives, One Hundred Twelfth Congress, second session /

Shipping list no. 2012-0243-P (pt. 1, 4), 2012-0266-P (pt. 2A), 2012-0267-P (pt. 2B), 2012-0250-P (pt. 3), 2013-0038 (pt. 5), 2013-0040-P (pt. 6), 2012-0298-P (pt. 7).

Departments of Labor, Health and Human Services, Education, and related agencies appropriations for 2014 : hearings before a subcommittee of the Committee on Appropriations, House of Representatives, One Hundred Thirteenth Congress, first session /

Mode of access: Internet.

The existence of evil spirits proved; and their agency, particularly in relation to the human race, explained and illustrated.

Mode of access: Internet.

The science of human behavior; biological and psychological foundations,

"Partial bibliography": p. 425-436.

USDA human nutrition research and education : report to Congress /

Mode of access: Internet.