bluuebunny/crossref_metadata_2025 huggingface dataset:
For White Men Only: The Socialist Party of America and Issues of Gender, Ethnicity and Race
Authors: Miller Sally M.
Published: November 2010

This article examines the experience of distinct groups within the Socialist Party of America, considering the period from its founding in 1901 to the eve of World War One. The Socialist Party, befitting its membership in the international socialist movement of that era, was committed to representing the workers of the world in the Marxist struggle to achieve political and economic equality within a cooperative commonwealth. Accordingly, variables of gender, ethnicity and race were subsumed within the Social Question. Ideally, workers in every capacity and of all backgrounds would march together under the Red Flag and create a new egalitarian society. While, indeed, many lawyers, ministers and others of the middle-class belonged to the Socialist Party and often held office within it, thereby derisively being likened by critics to an assemblage of dentists, the party was very much working class in composition, as shown by James R. Green and others.
“The Workingman's Bible”: Robert Blatchford'sMerrie England, Radical Literacy, and the Making of Debsian Socialism, 1895–1900
Authors: Martinek Jason D.
Published: November 2010

Turn-of-the-century socialists radicalized literacy. Unlike middle-class reformers, whose desire for mass literacy arose from the need for a hardworking, compliant workforce, socialists used it to undermine capitalism. Through their printed culture of dissent, they not only sought to transform individual lives, but an entire social system. They took up the task of using literacy to convert workers with a missionary zeal. Moral indignation fueled their crusade. In a nation of such wealth, they asked, why was it that so many industrious people did not have enough to provide themselves and their families with adequate food, clothing, and shelter? Their answer was that America's political and economic institutions had been corrupted by the nation's monied power. In their minds, only an enlightened, educated working-class could challenge the prerogatives of capital and make these institutions fully socially accountable to the people.
War and the Intellectuals: Bourne, Dewey, and the Fate of Pragmatism
Authors: Livingston James
Published: November 2010

My purpose here, apart from convincing you that John Dewey was quite possibly right about American entry into World War I, is to address the repression and mutilation of pragmatism by left-wing intellectuals in the twentieth century. These would seem to be very different purposes, but in fact they are the same. If we are to understand how pragmatism acquired its unsavory reputation among leftists everywhere, we must go back to 1917, when Randolph Bourne denounced not only Dewey's decision in favor of American entry but also pragmatism itself as the source of that decision. These almost ancient denunciations would not matter very much, except that they are repeated in every subsequent account of the American Left in World War I, and are recalled if not reiterated in every subsequent critique of pragmatism – they still determine our thinking about Dewey, about pragmatism, and about the war. Revisiting this primal scene allows us to ask why. It allows us to convert the following statement, which still serves as a left-wing credential, into a question: Dewey's support for American entry into the Great War demonstrates that pragmatism is a philosophy of acquiescence to “the existing fact,” a philosophy that must validate capitalism, accept imperialism, and repudiate socialism.
All Politics Are Local: Another Look at the 1890s
Authors: Argersinger Peter H.
Published: November 2010

Although rarely considered by historians, legislative and congressional apportionments were among the most important, absorbing, and contentious political issues of the late nineteenth century. Local, state, and national party leaders struggled to shape apportionments and thereby secure disproportionate influence for the counties, districts, and states their followers controlled. Gerrymanders, in turn, not only distorted representation but often incited a furious opposition, which disrupted legislative bodies, transformed political campaigns, and ultimately produced unprecedented judicial intervention. In surveying these overlooked developments, this essay points to important questions that historians must hereafter address.
Inventing “Little Italy”
Authors: Gabaccia Donna R.
Published: November 2010

Digitized texts open new methodologies for explorations of the history of ideas. This paper locates the invention of the term “Little Italy” in New York in the 1880s and explores its rapid spread through print and popular culture from police reporting to fictional portraits of slumming and then into adolescent dime novels and early film representations. New Yorkers invented “Little Italy” but they long disagreed with urban tourists about its exact location. Still, from the moment of its origin, both visitors and natives of New York associated Little Italy with entertainment, spectacle, and the search for “safe danger.” While the location of Little Italy changed over time, such associations with pleasure and crime have persisted, even as the neighborhood emptied of its immigrant residents.
Philadelphia's Lords of the Docks: Interracial Unionism Wobbly-Style
Authors: Cole Peter
Published: November 2010

In the early twentieth century, several thousand Philadelphia longshoremen organized themselves into a powerful, durable, and effective labor union. These men, who proudly belonged to the Industrial Workers of the World, proved willing and able to employ the Wobblies’ direct action tactics to improve their lives. Perhaps even more impressive is that Local 8 was one of the most, perhaps the most, racially inclusive union of its era. Few institutions of any sort at that time in America could claim to be more committed to interracial, multiethnic unionism than Local 8. For ideological and pragmatic reasons, Local 8 stood for racial and ethnic integration on the waterfront. Uniting a diverse workforce was essential to the union's success. Indeed, the union collapsed when Local 8 was split along racial lines. This article looks at the rise and fall of the Progressive Era's most integrated union.
‘An Actual Working Out of Internationalism’: Russian Politics, Zionism, and Lillian Wald's Ethnic Progressivism
Authors: Feld Marjorie N.
Published: November 2010

Students of the life of Lillian D. Wald (1867–1940) know her best as a Progressive activist. A trained nurse and advocate for East European Jewish immigrants on New York's Lower East Side, she founded Henry Street Settlement House there in 1893 and worked for state intervention in public health issues concerning women and children. Though she lived until 1940, historians have focused almost exclusively on her achievements before 1920: her founding of Henry Street, her key role in the formation of the Children's Bureau, her anti-militarism during World War I. This is not surprising, given that Wald' s rhetoric is that of a dyed-in-the-wool Progressive. She consistently cited her actions as in line with her universalist philosophy of human interdependence, which she referred to as “mutuality” and defined as a vision in which “no one class of people can be independent of the other”. Wald's mutuality echoes the Protestant social gospel movement's call for a “brotherhood of man” which inspired so many – including so many middle-class women – to work for various currents of Progressive reform.
“Not Designed Merely to Heal”: Women Reformers and the Emergence of Children's Hospitals
Authors: Sloane David
Published: November 2010

Children were a special concern of women reformers in the Gilded Age and Progressive Era. Assembling in federations and associations, women were, as Mary Odem has written, “especially active in efforts that aimed to protect women, children, and the home from the harmful effects of rapid urban growth and industrial capitalism.” Poor children were at risk due to industrial accidents, epidemics, and the stress and exhaustion of simply surviving in crowded tenements and polluted cities. Daphne Spain has suggested that women “saved the city” by starting political coalitions, improving neighborhood environments, and fighting for a wide range of protective legislation. Among those reforms was the nationwide movement to establish medical services for children. New pediatric wards and children's hospitals were intended to be places of comfort and cure as well as moral and spiritual education for the “little sufferers” and their parents.
Defective or Disabled?: Race, Medicine, and Eugenics in Progressive Era Virginia and Alabama
Authors: Dorr Gregory Michael
Published: November 2010

Something was menacing the South during the Progressive Era. Southern physicians located the threat in the “germ plasm,” the genes, of the region's inhabitants. Writing in a now-infamous 1893 “open letter” published in theVirginia Medical Monthly, Hunter Holmes McGuire, a Richmond physician and president of the American Medical Association, asked for “some scientific explanation of the sexual perversion in the negro of the present day.” McGuire's correspondent, Chicago physician G. Frank Lydston, replied that African-American men raped white women because of “[h]ereditary influences descending from the uncivilized ancestors of our negroes.” Lydston's solution to this problem was not lynching, but surgical castration which “prevents the criminal from perpetuating his kind.” Eight years later in Alabama, Dr. John E. Purdon opined, “It is a proved fact of experience that the inveterate criminal tends to propagates a race of criminals, and that the undeveloped or degraded nerve-tissue will duplicate itself in the next generation.” Dr. Purdon then declared, “Emasculation is the simplest and most perfect plan that can be adopted to secure the perfection of the race.” Twenty-three years later, in 1924, Harry Hamilton Laughlin testified in support of a Virginia law providing for the eugenic sterilization of the “shiftless, ignorant, and worthless class of anti-social whites of the South,” who allegedly created social problems for “normal” people. The multiplication of these “defective delinquents,” Laughlin and Virginia officials claimed, could only be controlled by restricting their procreation.
Culture Wars: The U.S. Art Lobby and Congressional Tariff Legislation during the Gilded Age and Progressive Era
Authors: May Robert E.
Published: November 2010

From 1883 to World War I, disputes over art tariffs roiled America's art community, drawing preeminent painters, sculptors, architects, and illustrators into national lobbying campaigns. This essay exposes artists’ agency in tariff politics, illuminates their ideologies, and explains congressional debates, legislation, and diplomacy regarding U.S. art schedules, while demonstrating how the art tariff imbroglio often challenged longstanding partisan patterns in Washington with respect to tariff protectionism. It also contributes to Atlantic world studies by exploring how artists’ anti-tariff positions derived from transoceanic systems of art pedagogy and exhibitions and by showing how protectionists (including a minority of artists) capitalized upon persistent popular stereotypes of national cultural inferiority. Finally, this essay argues that growing disparities of wealth and class sensitivities increasingly affected turn-of-the-century tariff discourse. Protectionists demanded punitive retribution against the international collecting activities of America's ostentatious plutocrats; free-art proponents craved tariff reforms for the didactic purpose of elevating popular taste through exposure to European masterworks.
“More Conscience Than Force”: U.S. Indian Inspector William Vandever, Grant's Peace Policy, and Protestant Whiteness
Authors: Anderson Douglas Firth
Published: November 2010

William Vandever (1817–1893) served as a U.S. Indian inspector from 1873 until early 1878. A lawyer by profession, Vandever had been a Republican congressman from Iowa and a Civil War officer. (Later, he would return to Congress, representing California.). While serving with the Indian Office, he became a critic of the militarization of federal Indian policy, so much so as to be reprimanded and not reappointed. His experience enables a reconsideration of President U.S. Grant's peace policy in at least two areas. First, as one of a new group of Office of Indian Affairs officials, Vandever provides a view of federal Indian policy from the middle level of the federal bureaucracy during the 1870s. His case especially illustrates his bureau's attempts to centralize civilian management of Indian reservations. Second, Vandever's policy criticisms, though they assumed white American “civilization” as normative, more immediately arose from his religious perspective. Although he lost his post, Vandever serves to highlight the privileged role of white Protestantism during Grant's peace policy. He exemplified a set of racialized religious sensibilities that were important at the time and that could be termed Protestant whiteness.
Resonant X-ray scattering study of perovskite-type vanadate RVO3
Authors: Nakao H. , Bizen D. , Shirane N. , Ikeuchi K. , Murakami Y. , Fujioka J. , Yasue T. , Miyasaka S. , Tokura Y.
Published: November 2010

The orbital ordering in perovskite-type vanadium oxides, RVO3 (R: rare earth), has been investigated by resonant X-ray scattering (RXS) near the V K-edge energy. The G-type orbital order, C-type orbital order and orbital disorder phases are elucidated on the basis of the azimuthal-angle and polarization dependence of the RXS signal reflecting the orbital ordering.
Inference of field‐scale fracture transmissivities in crystalline rock using flow log measurements
Authors: Frampton A. , Cvetkovic V.
Published: November 2010

Characterization of transmissivity for crystalline rock is conducted through simulation by conditioning against borehole flow rates obtained from high‐resolution, in situ field measurements during extraction pumping. Full three‐dimensional discrete fracture network simulations are carried out according to specifications obtained from site characterization data in a stochastic Monte Carlo setting. A novel method of conditioning is thereby introduced and applied using nonparametric comparison tests, which provide quantifiable measures of accuracy enabling evaluation of simulated results against field measurements. The assumption of a constitutive relationship (perfect correlation) between fracture size and transmissivity is adopted. The method is evaluated against both single and multiple realizations, various domain size, and fracture length configurations and shown to be robust for the cases considered. When the introduced method of conditioning is applied, transmissivity parameterization can be inferred to a narrow range with a quantifiable accuracy in terms of a probability value. Results indicate that elementary interpretation of transmissivity based on homogenization of a porous medium will generally underestimate transmissivity. Further implications on advective transport for natural flow conditions are briefly evaluated, indicating advective breakthrough times can be overestimated up to a factor of about 10 in the median.
Laryngeal Contribution to Cough
Authors: Pitts Teresa E. , Sapienza Christine M.
Published: November 2010

Cough is an important airway protection reflex. While cough involves shared anatomy with both the functions of swallowing and voice, its function is less understood by speech-language pathologists. The purpose of this article is to provide speech-language pathologists with a brief summary of the cough mechanism, its measurement, and its potential role in clinical evaluation and treatment.
Quasicylindrical description of a swirling light gas jet discharging into a heavier ambient gas
Authors: Gallardo-Ruiz J. M. , del Pino C. , Fernandez-Feria R.
Published: November 2010

The structure of an axisymmetric swirling gas jet of a light species discharging into an ambient of a heavier gas is analyzed using the quasicylindrical approximation of the compressible flow equations, with the main aim of describing the conditions for the onset of vortex breakdown. A self-similar solution valid in the mixing-layer close to the jet exit is found, which is used to start the numerical integration of the parabolic equations. For the computations, we consider the particular case of a swirling jet of hydrogen discharging into air. We characterize the critical swirl number for vortex breakdown as a function of the coflow velocity of the ambient gas, and compare it to the case of a homogeneous, single-species gas jet, discussing the physical differences found between the cases. We also consider the influence of the Mach number on the onset of vortex breakdown, and discuss the results in relation to the incompressible limit, finding that the swirl level for breakdown decreases as the Mach number increases.
Does viscosity describe the kinetic barrier for crystal growth from the liquidus to the glass transition?
Authors: Nascimento Marcio Luis Ferreira , Dutra Zanotto Edgar
Published: November 2010

An analysis of the kinetic coefficient of crystal growth Ukin, recently proposed by Ediger et al. [J. Chem. Phys. 128, 034709 (2008)], indicates that the Stokes–Einstein/Eyring (SE/E) equation does not describe the diffusion process controlling crystal growth rates in fragile glass-forming liquids. Ukin was defined using the normal growth model and tested for crystal data for inorganic and organic liquids covering a viscosity range of about 104–1012 Pa s. Here, we revisit their interesting finding considering two other models: the screw dislocation (SD) and the two-dimensional surface nucleated (2D) growth models for nine undercooled oxide liquids, in a wider temperature range, from slightly below the melting point down to the glass transition region Tg, thus covering a wider viscosity range: 101–1013 Pa s. We then propose and use normalized kinetic coefficients (Mkin) for the SD and 2D growth models. These new kinetic coefficients restore the ability of viscosity to describe the transport part of crystal growth rates (Mkin∼1/η and ξ∼1) from low to moderate viscosities (η<106 Pa s), and thus the SE/E equation works well in this viscosity range for all systems tested. For strong glasses, the SE/E equation works well from low to high viscosities, from the melting point down to Tg! However, for at least three fragile liquids, diopside (kink at 1.08Tg, η=1.6×108 Pa s), lead metasilicate (kink at 1.14Tg, η=4.3×106 Pa s), and lithium disilicate (kink at 1.11Tg, η=1.6×108 Pa s), there are clear signs of a breakdown of the SE/E equation at these higher viscosities. Our results corroborate the findings of Ediger et al. and demonstrate that viscosity data cannot be used to describe the transport part of the crystal growth (via the SE/E equation) in fragile glasses in the neighborhood of Tg.
Shock-wave solutions in two-layer channel flow. I. One-dimensional flows
Authors: Mavromoustaki A. , Matar O. K. , Craster R. V.
Published: November 2010

We study the dynamics of an interface separating two immiscible layers in an inclined channel. Lubrication theory is used to derive an evolution equation for the interface position that models the two-dimensional flow in both co- and countercurrent configurations. This equation is parameterized by viscosity and density ratios, and a total dimensionless flow rate; the system is further characterized by the height of the interface at the channel inlet and outlet, which are treated as additional parameters. In the present work, which corresponds to part I of a two-part paper, we focus on one-dimensional flows. We use an entropy-flux analysis to delineate the existence of various types of shocklike solutions, which include compressive Lax shocks, pairs of Lax and under-compressive shocks, and rarefaction waves. Flows characterized by unstably stratified layers are accompanied by the formation of propagating, large-amplitude interfacial waves, which are not shocklike in nature. The results of our transient numerical simulations accord with our analytical predictions and elucidate the mechanisms underlying spatio-temporal development of the various types of waves; the stability of these waves to spanwise perturbations is investigated in part II of this work.
Study of the isotropic to smectic-A phase transition in liquid crystal and acetone binary mixtures
Authors: Sigdel Krishna P. , Iannacchione Germano S.
Published: November 2010

The first-order transition from the isotropic (I) to smectic-A (Sm A) phase in the liquid crystal 4-cyano-4′-decylbiphenyl (10CB) doped with the polar solvent acetone (ace) has been studied as a function of solvent concentration by high-resolution ac-calorimetry. Heating and cooling scans were performed for miscible 10CB+ace samples having acetone mole fractions from xace=0.05 (1 wt %) to 0.36 (10%) over a wide temperature range from 310 to 327 K. Two distinct first-order phase transition features are observed in the mixture whereas there is only one transition (I-Sm A) in the pure 10CB for that particular temperature range. Both calorimetric features reproduce on repeated heating and cooling scans and evolve with increasing xace with the high-temperature feature relatively stable in temperature but reduced in size while the low-temperature feature shifts dramatically to lower temperature and exhibits increased dispersion. The coexistence region increases for the low-temperature feature but remains fairly constant for the high-temperature feature as a function of xace. Polarizing optical microscopy supports the identification of a smectic phase below the high-temperature heat capacity signature indicating that the low-temperature feature represents an injected smectic-smectic phase transition. These effects may be the consequence of screening the intermolecular potential of the liquid crystals by the solvent that stabilizes a weak smectic phase intermediate of the isotropic and pure smectic-A.
Classical photodissociation dynamics with Bohr quantization
Authors: Bonnet L.
Published: November 2010

The standard classical expression of the state-resolved photodissociation cross section is not consistent with an efficient Bohr quantization of product internal motions. A new and strictly equivalent expression not suffering from this drawback is proposed. This expression opens the way to more realistic classical simulations of direct polyatomic photodissociations in the quantum regime where only a few states are available to the products.
Density functional study of magnetic properties in Zn-doped SnO2
Authors: Wei Wei , Dai Ying , Guo Meng , Lai Kangrong , Huang Baibiao
Published: November 2010

Magnetic properties in Zn-doped SnO2 are examined through the first-principles electronic structure calculations based on density functional theory. Our results reveal that Zn-doping induces localized spin magnetic moments primarily on the first coordination shell of O atoms surrounding the Zn atom as well on Zn atom with total magnetic moment of 1.47 μB per supercell. Holes localized on O atoms in ZnO6 are polarized with the same spin orientation as that of the dopant. Ferromagnetic coupling between Zn ions in Zn-doped SnO2 is attributed to the hole-mediated p-d exchange coupling interaction. With respect to native defects in Zinc-doped SnO2, formation of oxygen vacancy (VO) is suppressed whereas formation of tin vacancy (VSn) is facilitated due to Zn-doping. It is found that the observed ferromagnetism in Zn-doped SnO2 mainly originates from the Zn dopant rather than exclusively the formation of VSn.
Using the friendship paradox to sample a social network
Authors: Wilson Mark
Published: November 2010

When applied to random nodes in a network, the statement “Your friends have more friends than you do” has predictive power.
Optical manipulation of light-absorbing particles takes to the air
Authors: Smart Ashley G.
Published: November 2010

An effect known as photophoresis sends carbon-coated microspheres traveling unprecedented distances inside a beam of light.
041.Activin A: from reproductive factor to inflammatory cytokine
Authors: Phillips D. J. , Jones K. L. , O'Connor A. E. , Wilson K. M. , Patella S. , Sebire K. , De Kretser D. M.
Published: November 2010

Activin A was originally isolated and characterised as a reproductive feedback regulator of follicle-stimulating hormone. While potent paracrine networks involving activin and its binding protein, follistatin, are present in the gonad and pituitary, recent focus has been on emerging roles in a number of other systems, such as erythropoiesis, neuronal survival, embryonic development and inflammatory processes. The latter relatively new property was first suggested by us when follistatin in the circulation was elevated in sheep undergoing surgical trauma. We have since focussed on a model of acute inflammatory challenge using the bacterial cell wall component lipopolysaccharide (LPS) or endotoxin. This has highlighted that the release of activin into the bloodstream occurs extremely rapidly, within about 50�min. The response appears to be biphasic and precedes or is at least coincident with the release of a number of key pro-inflammatory cytokines, such as tumour necrosis factor α and interleukin-6. The mechanisms of this release are still being delineated, but it is fever- and prostaglandin-independent, and largely unaffected by blocking other key cytokine responses. Nevertheless, it is directly downstream of the LPS receptor and its activation pathway. Importantly, activin's property as an inflammatory cytokine appears to be borne out in a number of clinical inflammatory syndromes such as septicemia, suggesting that it is a hitherto undescribed component of the organism's innate immune response to infection.
125. Differential regulation of inhibin binding via betaglycan expression in several mouse cell lines
Authors: Farnworth P. G. , Wang Y. , Ooi G. T. , Findlay J. K.
Published: November 2010

Inhibin A, a member of the transforming growth factor (TGF)-β superfamily, binds to mouse adrenocortical (AC), Leydig (TM3) and Sertoli (TM4) cell lines with high affinity via at least eight membrane protein species, two of which are forms of betaglycan. Inhibin A has been proposed to inhibit the actions of activin and BMP by sequestering their type II receptors in high affinity complexes with betaglycan (1). We previously found that BMPs appear to counteract inhibin action in AC cells by selectively suppressing the expression of endogenous betaglycan, consequently reducing inhibin binding. In the present studies, we have examined how factors that stimulate betaglycan expression in other systems modify the binding of radiolabelled inhibin A at the surface of AC, TM3 and TM4 cells.
AC, TM3 and TM4 cells were treated overnight with glucocorticoid, membrane-permeable cAMP analogue or retinoic acid, after which the levels of betaglycan mRNA, corrected for GAPDH content, were measured using real-time RT-PCR, and [125I]inhibin A binding was determined. Treatment of AC cell cultures with 8Br-cAMP (1�mM), glucocorticoid (RU28362, 100�nM) or retinoic acid (30�mM) increased betaglycan mRNA levels 120-150%, and increased subsequent inhibin A binding to 146���12, 132���13 and 125���18% of control (mean���SD, n = 6–12). The glucocorticoid and cAMP treatments also increased inhibin binding to TM3 and TM4 cells by similar amounts, but retinoic acid was less effective. Affinity labelled protein species of deduced sizes 115 and >170�kDa, consistent in size with betaglycan forms, were the primary target for stimulation by these agents, whereas species of 65 and 75�kDa were selectively increased by retinoic acid in the AC cells.
In summary, glucocorticoids, retinoids and hormones that stimulate cAMP production may increase the expression of betaglycan in inhibin target cells, increase their binding of inhibin, and thereby promote inhibin action. These studies confirm that betaglycan is a primary determinant of inhibin binding and action. The protein species other than betaglycan that are selectively upregulated by retinoic acid in AC cells are yet to be identified. Funded by the NH&MRC of Australia (RegKey 241000 & 198705).
(1) Wiater & Vale (2003) J. Biol. Chem. 278, 7934.
212.The molecular basis of epididymal sperm maturation
Authors: Nixon B. , Harman A. J. , Asquith K. L. , Aitken R. J.
Published: November 2010

The mammalian epididymis represents the site where functionally incompetent spermatozoa originating from the testes undergo their final maturation enabling them to engage in the complex cascade of sperm-egg interactions that culminate in fertilization. The extent to which this process is actively driven by the epididymis or reflects properties intrinsic to the gamete remains largely unknown. However, recent studies within our laboratory have demonstrated that sperm transit through region 4 (corpus) of the mouse epididymis is associated with an acquired ability to exhibit coordinated movement, capacitate and engage in sperm-zona binding. Furthermore, we have demonstrated that immature mouse sperm recovered from region 3 (caput) of the epididymis are able to display similar attributes following brief co-culture with region 4 epididymal plasma. Interestingly, immunohistological studies of the epididymal lumen within region 4 have revealed the presence of a number of dense bodies containing the molecular chaperones, heat shock protein 60 (HSP60) and endoplasmin (GRP94). Although the nature and origin of such inclusions remains to be resolved, these collective findings raise the intriguing possibility that the molecular chaperones are involved in the delivery of critical signaling molecules to the surface of spermatozoa. In light of this data, we have commenced a proteomic analysis of region 4 epididymal fluid with a view to identifying proteins which interact with the chaperones, HSP60 and GRP94. Such studies will provide an extremely important insight into the molecular basis of sperm maturation.
220.TGFβ1 deficient mice exhibit impaired follicle growth and luteal maintenance
Authors: Robker R. L. , Ingman W. V. , Robertson S. A.
Published: November 2010

Transforming Growth Factor β1 (TGFβ1) is essential for normal female reproduction. Mice with a targeted deletion in the TGFβ1 gene (TGFβ1–/–) have severely impaired fertility with pregnancy occurring in <25% of mated females. TGFβ1 is implicated in several aspects of ovarian function, including potentiation of granulosa cell proliferation and suppression of luteal cell apoptosis. Our initial observations indicate that estrous cycling is disrupted in TGFβ1–/– mice and that ovulation rate is reduced. To further investigate how impaired ovarian function contributes to the infertility of TGFβ1–/– mice, ovaries were isolated from TGFβ1+/+ and TGFβ1–/– littermates at proestrus and fixed and sectioned for examination of follicle morphology and growth. BrdU labelling was performed to detect granulosa cell proliferation and blood samples were obtained for analysis of gonadotrophins and ovarian steroid hormones. Histological examination showed that ovaries from TGFβ1–/– mice were smaller than those of TGF–1+/+ mice, however large antral follicles were observed, indicating that TGFβ1 is not essential for granulosa cell proliferation. Compared to TGFβ1+/+ ovaries however, there were fewer antral follicles and only rare corpora lutea. Interestingly, in some cases there were large numbers of macrophages surrounding small follicles suggesting increased follicular atresia and/or altered macrophage activity in the TGFβ1–/– ovaries. Ovaries and serum were also isolated from females at d4 post-coital for assessment of corpora lutea morphology. TGFβ1–/– ovaries weighed less and had fewer corpora lutea than TGFβ1+/+ ovaries. TGFβ1–/– corpora lutea also contained increased numbers of apoptotic cells and infiltrating macrophages indicative of premature luteal regression. Circulating progesterone levels were reduced in TGFβ1–/– females, as was progesterone production per corpus luteum further indicating a functional defect in luteal maintenance. Cumulatively these observations show that TGFβ1 has essential roles in regulation of ovarian macrophage populations, in normal follicular development and in the generation, maintenance and steroidogenic function of corpora lutea.
227.An inhibitor of leukemia inhibitory factor signalling blocks embryo implantation in the mouse
Authors: Dimitriadis E. , Stoikos C. , Baca M. , Fairlie W. , Uboldi A. D. , McCoubrie J. E. , Salamonsen L. A.
Published: November 2010

Embryo implantation is a critical step in the establishment of pregnancy. Endometrial leukemia inhibitory factor (LIF) is essential for embryo implantation in the mouse (1). Uterine LIF is expressed in the luminal epithelium on Day 3 of pregnancy (D3) (D0�=�day of plug detection) and signals via activation of signal transducer and activator of transcription (Stat) 3 (2). We examined the effect of a novel LIF signalling inhibitor on the phosphorylation (p) of Stat3 during early pregnancy and on embryo implantation in the mouse. We injected LIF inhibitor into one uterine horn and PBS into the other uterine horn of the mouse at D3 and examined the effect on pStat3 immunostaining in the luminal epithelium between 30 and 360�min later. We found no immunoreactive pStat3 in luminal epithelium following treatment with LIF inhibitor at 60 and 90�min but variable staining at other time points. The PBS-treated uterine horn showed intense immunostaining at all times. LIF inhibitor (1mg/kg body weight per day) or PBS was administered to mice (a) subcutaneously, (b) intraperitoneally, at 8-hourly intervals for 3�days from D2, or (c) continuously into the peritoneal cavity via Alzet pumps from D2. No effect was seen on implantation at D6. When LIF antagonist (3.5mg/kg/day) or PBS were administered by Alzet pumps from D2 together with ip injections, 4-hourly from D3 for 36�h, there were no implantation sites in the uteri of treated mice (n�=�5) while the control mice (n�=�4) had 3.6���0.5�sites (P�<�0.001). Histologically, the uteri of the treated mice resembled non-pregnant uterus, while the control uterus resembled post-implantation uterus. The results demonstrate that treatment of mice during early pregnancy with a novel LIF inhibitor blocks LIF action in vivo and embryo implantation. This knowledge is important for development of novel contraceptives.
(1) Stewart, C. L., Kaspar, P., Brunet, L. J., Bhatt, H., Gadi, I., Kontgen, F., Abbondanzo, S. J. (1992) Nature 359, 76–79. (2) Cheng, J. G., Chen, J. R., Hernandez, L., Alvord, W. G., Stewart, C. L. (2001) Proc. Natl Acad. Sci. USA 98, 8680–8685.
237.Insulin receptor expression in mouse preimplantation embryos
Authors: Hung F. , Pantaleon M. , Kaye P. L.
Published: November 2010

The insulin receptor (IR) mediates the actions of insulin and insulin-like growth factors (IGF-I and II). Two IR isoforms result from alternate splicing of exon 11, IR-A (without exon 11) and IR-B (with exon 11). Exon 11 is 36 bp and encodes 12 amino acids (717-729) in the COOH-terminus of the IR alpha-subunit. IR-A has higher binding affinity for insulin and IGF-II than IR-B. Interestingly, IR-A is predominantly expressed in fetal tissues, adult spleen, brain and tumours, where its expression has been associated with an undifferentiated state. IR-B is expressed primarily in classical insulin sensitive tissues such as adult liver and muscle. In contrast to other species, no evidence existed for IR expression prior to compaction in mice. However, both insulin and IGF-II have been shown to stimulate growth in cleaving embryos (Kaye 1997). We returned to this question using primers flanking exon 11 to detect IR-A and IR-B mRNA. Sequencing confirmed the identity of the amplified PCR products. IR-B mRNA was present in zygotes and all stages to blastocyst, IR-A mRNA was only detected in compacted morulae. Immunofluorescent confocal microsopy using a rabbit polyclonal antibody (against amino acids 128-205 of alpha-subunit) that recognizes both isoforms revealed nuclear IR localization in zygotes and cleavage stage embryos including morulae. The results demonstrate that mice like other species expresses IR throughout preimplantation development in contrast with earlier studies. Nuclear localisation of IR has been observed in hepatocytes and adipocytes and may be associated with transcriptional regulation. IR-A expression in compacted morulae may reflect the change in metabolism that occurs at this time and requires further study.
(1) Kaye, P. L. (1997) Preimplantation growth factor physiology. Rev. Reprod. 2, 121–127.
249.Activin A upregulates endometrial metalloproteases: potential mechanisms for promotion of decidualisation and implantation
Authors: Jones R. L. , Paiva P. , Kaitu'u T. J. , Salamonsen L. A.
Published: November 2010

Activin and inhibin subunits are co-expressed by human endometrial epithelial and decidualised stromal cells. Activin A is a potent stimulator of decidualisation in vitro, but the mechanisms are unknown. Matrix metalloproteases (MMPs) are known to be important during decidualisation, as administration of a broad spectrum MMP inhibitor in the rat results in reduced decidualisation. Transforming Growth Factor(TGF)-βs are closely related to activins and inhibit MMP production in endometrial epithelial cells. We hypothesised that activins regulate MMP production during decidualisation and/or trophoblast invasion. Epithelial and stromal cells were isolated from human endometrium and treated for 24 h with activin, inhibin, activin/inhibin, and follistatin. Media were collected and subjected to gelatin and caesin zymography. In epithelial cells, activin A stimulated the expression of latent forms of MMPs-1, -2, -7 and -9, and increased formation of active forms of MMPs-2 and -7. Cotreatment with inhibin prevented this stimulation, whilst inhibin alone completely inhibited MMP production. Treatment with follistatin treatment reduced MMP levels. Similar regulation was seen in stromal cells for MMPs-1, -2 and -9. These data show that activin stimulates the production and activation of MMPs in both endometrial cells, and that inhibin is a potent inhibitor. It is interesting that activin is acting in an opposing manner to TGF-β, indicating that these two closely related proteins have divergent signalling pathways in endometrial cells. Further, this is the first demonstration of a role for inhibin in regulating MMPs and indeed for inhibin action in the endometrium. These findings are of potential importance in understanding regulation of MMPs in the peri-implantation endometrium. Activin is the predominant dimer produced by decidual and epithelial cells, where it may be promoting decidualisation though enhancing MMP production and activation. Furthermore, activin secretion by invasive cytotrophoblasts may stimulate focal decidual MMP production promoting their invasion during embryo implantation.
256.Development of a label retaining cell method to identify stem cells in mouse endometrium
Authors: Chan R. W. , Gargett C. E.
Published: November 2010

Epithelial and stromal stem/progenitor cells have been demonstrated in human endometrium by their clonogenic activity (1). To understand the regulation and growth kinetics of endometrial stem cells (SCs), an animal model is being established using the label retaining cell (LRC) approach. Tissue SCs are quiescent and will retain a DNA synthesis label (BrdU), while the label is diluted out in more mature dividing cells during a chase period. Those cells retaining the label over long period of time are LRCs and have been identified as SCs (2). An optimum time to initially label the majority of the cells is critical. We hypothesised that during mouse endometrial development, there is a growth window for maximum labelling of epithelial and stromal cells. The aim of this study was to determine the optimum age and length of time for maximum endometrial BrdU labelling. Two stages of endometrial growth were exploited: (1) postnatal growth before gland development (3 days old – P3); and (2) oestrogen induced growth in ovariectomised prepubertal mice (4–5 weeks old). C57/CBA P3 mice received subcutaneous BrdU injections twice daily for 3 consecutive days. Four to five-week-old ovariectomised mice received BrdU filled mini-osmotic pumps implanted subcutaneously for 7 and 14 days, with daily oestrogen injections. Mice were killed 4 h after the last treatment, uteri collected, and BrdU labelled cells were detected by immunohistochemistry. Results are shown in the table below (see PDF file).
The data shows that maximum labelling for epithelial and stromal cells was achieved for P3 mice, possibly due to greater endometrial growth of the Müllerian duct compared to oestrogen induced endometrial growth following ovariectomy. Chase experiments are currently in progress on P3 labelled mice to identify the location of LRCs in mouse endometrial glands and stroma.
(1) Chan RW, Schwab KE, Gargett CE (2004) Biol. Reprod. 70, in press. (2) Morris RJ, Potten CS (1994) Cell Prolif. 27, 279–289.
258.Natural killer cell activity in the adult rat testis
Authors: Aridi D. , Hutchinson P. , Hedger M.
Published: November 2010

Immune privilege of the testis is due to an inability to recognise and respond to antigens within the testis, and implies that innate immunity against tumours and infections, such as that afforded by natural killer (NK) cells, may be important. Our previous studies established the existence of cells that express NK markers in the rat testis, but the functional activity of these cells has not been assessed previously. Testicular interstitial cells were collected from adult male Sprague-Dawley rats by mechanical dissociation, and macrophages were removed by adherence depletion. These interstitial cell preparations contained on average 3% cells expressing the NK cell marker, CD161. The interstitial cells (effector cells) were labelled with 5-(6)-carboxy-fluorescein succinimydyl ester (CFSE), and cultured at various effector�:�target cell ratios with YAC-1 tumour cells (target cells) overnight. The combined cells were labelled with 7-amino actinomycin D (7-AAD), which stains the nuclei of cells undergoing apoptosis. Cells were fixed with paraformaldehyde and analysed by flow cytometry. The proportion of target (CSFE negative) cells expressing 7-AAD staining (mean���SD, n�=�4 experiments) increased as the effector�:�target cell ratio was increased: 3.5���3.6% (no. effector cells; background), 24���7.0% (effector�:�target cell ratio, 2�:�1), 25.8���11.7% (7�:�1), 41.8���17.0% (20�:�1), and 71.7���12.7% (60�:�1). These data indicate that functional NK cells are present in the rat testicular interstitial cell preparations, and may contribute to innate immune protection in the rat testis.
275.Calmodulin-dependent nuclear import pathway of the testis-determining factor SRY
Authors: Kaur G. , Delluc-Clavieres A. , Poon I. , Jans D. A.
Published: November 2010

Modulation of the nuclear entry of transcription factors (TFs) and chromatin components is a means by which eukaryotic cells can regulate gene expression in response to extracellular signals and the cell cycle during differentiation and development. TFs and chromatin components access the nucleus through nuclear localisation sequences (NLSs), which mediate interaction with components of the cellular nuclear import machinery, such as members of the importin superfamily. The Ca2+-binding protein calmodulin (CaM ) has previously been shown to bind at or near NLSs in several nuclear-localising proteins that have important roles in testis development including the Y chromosome-encoded HMG-domain-carrying chromatin remodelling factor SRY, and related factor SOX9, both of which are key regulators of gonadal development. SRY function in the nucleus of somatic cells of the fetal gonad, in particular, is essential for development of a testis in males. Here we present new findings implicating a role for CaM in modulating SRY nuclear accumulation, whereby treatment of transfected cells with CaM antagonists significantly reduces nuclear accumulation of green fluorescent protein (GFP)-fusion proteins encoding either full length SRY or the SRY HMG domain alone. An in vitro nuclear transport assay using bacterially expressed fluorescent proteins showed similar results, with native gel electrophoresis/fluorimaging and fluorescence polarisation assays, indicating direct binding of CaM to the SRY HMG domain in Ca2+-dependent fashion. Since clinical mutations resulting in sex reversal occur within SRY's CaM-binding NLS, these results may shed new insight into CaM-dependent pathways of nuclear protein import, and how this may relate to testis development.
286.Nuclear factor κB downregulation in human T-cells is essential for the maintenance of the cytokine profile required for pregnancy success
Authors: McCracken S. A. , Gallery E. D.M. , Morris J. M.
Published: November 2010

Human pregnancy is associated with a shift away from Th-1 type and a bias towards Th-2 type immune responses. The molecular mechanisms that regulate this are unknown. We assessed the expression and activity of Nuclear Factor (NF)-κB, a transcription factor that plays a central role in regulating immune responses. Nuclear and cytoplasmic fractions were prepared from isolated T-cells from non-pregnant and pregnant females and subjected to Western blotting to assess NF-κB and its' inhibitors IκBα and β expression. NF-κB activity in nuclear extracts was determined by Electrophoretic Mobility Shift Assays. Isolated T-cells were pre-incubated with/without the NF-κB translocation inhibitor SN50 and subsequently stimulated with PMA/ionomycin in the presence of the protein transport inhibitor Brefeldin A. Cytokine production was determined using flow cytometry.
Our results demonstrated high levels of immunoreactive NF-κB (p65) in all nuclear fractions of T-cells from non-pregnant females. In contrast, low levels of p65 were detected in nuclear fractions of T-cells from pregnant females. Levels of IκBα and β were also higher in cytoplasmic fractions of T-cells from non-pregnant than from pregnant females. The reduction in p65 levels in pregnancy was consistent with reduced levels of active NF-κB in T-cells from pregnant relative to non-pregnant females. Stimulation of T-cells from non-pregnant females with PMA/ionomycin resulted in IκBα degradation, p65 translocation and subsequent production of Th-1 cytokines IFN-β and IL-2. In contrast, PMA stimulation had no effect on NF-κB activity in T-cells from pregnant females and a reduced effect on IFN-β and IL-2 production. In the presence of SN50, IFN-β and IL-2 production by T-cells from non-pregnant females was attenuated demonstrating a specific role for NF-κB in the production of these Th-1 cytokines. We can therefore conclude that, specific down-regulation of NF-κB in T-cells in pregnancy is an essential requirement for maintaining the cytokine profile necessary for pregnancy success.