bluuebunny/crossref_metadata_2025 huggingface dataset:
Finding Meaning and Purpose in Boundaryless Careers: A Framework for Study and Practice
Authors: Lips-Wiersma Marjo , Mcmorland Judith
Published: March 2006

This article addresses a paradox in current boundaryless career theories. These theories span the boundaries of employment settings and occupations in response to changing social and economic conditions. In this climate, boundaryless career advocates claim that individuals need to be concerned with subjective career determinants, such as meaning and purpose, and objective career determinants, such as remaining employable. Such advocacy, however, does not adequately acknowledge that the very premise on which boundaryless career theory is built, namely that career choice is strongly driven by market forces, may counter one’s very ability to find meaning and purpose. The authors reintroduce the notion of vocation and identify four aspects through which the concept of vocation may be expanded: animation, dedication, evaluation, and coherence. They suggest that the discourses of career and vocation can complement one another to enable the individual to discover or maintain a sense of purpose in a boundaryless career climate.
Validation of the Body in Psychotherapy
Authors: Leijssen Mia
Published: March 2006

Psychotherapists can improve verbal psychotherapy by adding a bodily perspective. Different approaches can be situated on a continuum from verbal to nonverbal, and body-oriented interventions can be directed to different aspects of the body. The body as sensed from inside is one source of information. This is different from working with the body as perceived from outside and paying attention to nonverbal communication. In the next stage, major methods are working with the body in action and in movement and other nonverbal expressions. At the end of the continuum, attention goes to touching the body. Different ways of validating the body in psychotherapy are illustrated with clinical vignettes. Effects of body-oriented interventions on the client’s process are greater awareness, engagement in the present, deepening of experience, opening the body memory, cathartic release, resolving blocks, and exploring new possibilities.
The Costatement: Objective Evidence for a Science of Subjectivity
Authors: Reeves Ralph M.
Published: March 2006

Sometimes, in conversation, there occurs a particular kind of statement that sheds unusual light on the basic processes by which we affect and are affected by one another. These “costatements” have apparently not been previously noticed. The author offers several examples of them from his own practice and then explores some of their implications. These include their usefulness in monitoring the therapeutic cooperation, registering the effectiveness of the input of each partner to the other, indicating the degree to which the cocreation of their experience is actually discernible, and pointing to the occurrence of peaks of mutuality. The author ponders the bipersonal field out of which these statements arise, and the intriguing question of the role of spirit in such a field.
Social Construction as an Ethics of Infinitude: Reply to Brinkmann
Authors: Gergen Kenneth J.
Published: March 2006

This article responds to Svend Brinkmann’s (2005) critique of social constructionism, and most particularly to his charges that constructionist ideas contribute to an ethos of consumerism, ultimately related, as he sees it, to an ethics of infinitude. For Brinkmann, such an ethic denies the unconstructed facts of interdependency and mortality. These realities, for Brinkmann, are essential to any moral foundations at all. As I attempt to demonstrate, consumerism is one among many potential implications of constructionist ideas; however, the more demanding question is how to sustain concepts of trust and commitment in a globalizing world of multiple relationships. I further challenge Brinkmann’s characterization of constructionism as individualist and raise the possibility that all attempts to generate ethical foundations may be inimical to human life.
Divorce Motives in a Period of Rising Divorce
Authors: de Graaf Paul M. , Kalmijn Matthijs
Published: March 2006

Using survey data on 1,718 ever-divorced men and women in the Netherlands, the authors describe the motives people give for their divorce. The authors distinguish motives regarding three types of issues: relational issues, behavioral problems, and problems about work and the division of labor. They observe three important trends: the normalization of divorce, the psychologization of relationships, and the emancipation of women. First, severe divorce motives (e.g., violence and infidelity) have become less important. The authors interpret this finding in terms of a threshold hypothesis: When the threshold for divorce is higher, marriages that end in divorce will be more problematic. Second, there has been a trend toward more relational and psychological motives, particularly among women. Third, problems in the realm of work and household labor have become more important motives for a divorce. This is consistent with the increase in emancipatory attitudes in the past decades.
Wives’ Employment and Spouses’ Marital Happiness
Authors: Schoen Robert , Rogers Stacy J. , Amato Paul R.
Published: March 2006

The authors investigate the direction of the relationship between marital happiness and wives’ full-time employment using the 1987 to 1988 and 1992 to 1994 waves of the National Survey of Families and Households. First, the authors predict change in wives’ employment between the two waves using marital happiness and other Time 1 characteristics. The results show that shifting into full-time employment is more likely for unhappily married than for happily married wives. Second, they examine how changes in wives’ employment between Times 1 and 2 influence marital stability and changes in marital happiness. The authors find that contrary to frequently invoked social and economic theories, wives’ full-time employment is associated with greater marital stability. Nonetheless, changes in wives’ employment have no significant effect on how marital quality changes between Times 1 and 2.
Anxiety and Psychosocial Concerns in Alopecia Areata
Authors: Elkin T. David , Hilker Kellie A. , Drabman Ronald S.
Published: March 2006

Alopecia areata (AA), or hair loss on the scalp and body, affects approximately 1 in 1,000 people. Psychological distress has been hypothesized as a determining factor of AA. Researchers evaluating psychological disorders in the development and course of AA have found equivocal results. Furthermore, few studies have evaluated parental distress in relation to childhood AA. Case studies have shown the benefit of behavioral techniques for adults with AA; however, no studies have examined the role of behavioral interventions on child and parent coping abilities in AA. This brief case report will discuss the treatment of child-reported and parent-reported psychological distress in a child with AA. Significant reductions in child anxiety and social concerns were found over the course of treatment. This case report provides support for the use of behavioral techniques for the treatment of psychological distress experienced secondary to a child's diagnosis of AA.
A Cognitive-Behavioral, Biofeedback-Assisted Relaxation Treatment for Panic Disorder With Agoraphobia
Authors: Goodwin Elizabeth A. , Montgomery Doil D.
Published: March 2006

A 39-year-old man diagnosed with panic disorder with agoraphobia, comorbid with depression, was treated successfully with cognitive-behavioral, biofeedback-assisted relaxation therapy. Treatment consisted of 22 individual sessions over the course of 10 months. Of those, 18 sessions included biofeedback with electrodermal response as the primarymodality and finger temperature monitored as a general index of relaxation. The core interventions utilized during treatment included psychophysiological monitoring, relaxation techniques, cognitive restructuring, and exposure to feared cues. Results were cessation of panic attacks, full remission of agoraphobia, and alleviation of depressive symptoms. Assessments used during the course of treatment included the Beck Anxiety Inventory, State-Trait Anxiety Inventory Form Y-2, Agoraphobic Cognitions and Body Sensations Questionnaires, and the Beck Depression Inventory-II. A treatment formulation that combined the most effective interventions with cognitive-behavioral underpinnings is discussed along with recommendations for practice.
Integrative Strategies for Treating Internet Sexuality
Authors: Chirban John T.
Published: March 2006

This article introduces strategies for deepening the understanding of paraphilias and the therapeutic process of treating them. The case study explores a young man's compulsive sexual fantasies over the Internet, his compulsive masturbation, and his pedophilic and paraphilic fantasies as adaptations from early developmental conflicts. A therapeutic process supports this young man's understanding of his rejection of himself and promotes acceptance of developmentally appropriate and integrated sexual expression. In particular, this case discussion addresses three aspects in them ultiplicity of one's sexuality: the sexual self, the “voices” of sexuality, and the “dance” of sexuality. Additionally, management of transference and countertransference are discussed as means of strengthening the therapeutic alliance.
Changes in the Interpersonal Characteristics of a Client Who “Recovered” From Anxiety and Depression Following Interactionist Interpersonal Therapy
Authors: Shields Candyce , Murrell Stanley A. , Salsman Nicholas
Published: March 2006

This therapeutic approach assumes that for successful outcome, changes in clients' interpersonal patterns are necessary. This client, “Helen,” presented with symptoms of anxiety and depression. After 45 sessions, she had made clinically significant improvement in these symptoms. Based on our case formulation, specific interpersonal characteristics were identified as targets for change (i.e., to move from being hostile and distant as her predominant interpersonalpatternto beingmore affiliative and assertive). Onthe basis of scores on three different interpersonal circumplex instruments taken over the course of the 45 sessions, the changes were consistently in line with the targeted interpersonal characteristics. The therapist was able to respond to Helen's hostile, distant pattern with non-controlling friendly acceptance and with a focus on her internal experiences, which promoted a strong therapeutic bond. Contributions to the outcome of this specific approach in conjunction with common factors are discussed.
The SCL relative LYL-1 is required for fetal and adult hematopoietic stem cell function and B-cell differentiation
Authors: Capron Claude , Lécluse Yann , Kaushik Anna Lila , Foudi Adlen , Lacout Catherine , Sekkai Dalila , Godin Isabelle , Albagli Olivier , Poullion Isabelle , Svinartchouk Fedor , Schanze Elisabeth , Vainchenker William , Sablitzky Fred , Bennaceur-Griscelli Annelise , Duménil Dominique
Published: March 2006

Hematopoietic stem cells (HSCs) arise, self-renew, or give rise to all hematopoietic lineages through the effects of transcription factors activated by signaling cascades. Lyl-1 encodes a transcription factor containing a basic helix-hoop-helix (bHLH) motif closely related to scl/tal, which controls numerous decisions in embryonic and adult hematopoiesis. We report here that Lyl-1 null mice are viable and display normal blood cell counts, except for a reduced number of B cells resulting from a partial block after the pro-B stage. Nevertheless, the deletion of Lyl-1 results in a diminution in the frequency of immature progenitors (Lin–, CD34–, sca-1+, c-kit+ [LSK], and LSK-side population [LSK-SP]) and in S12 colony-forming unit (CFU-S12) and long-term culture-initiating cell (LTC-IC) content in embryonic day 14 fetal liver (E14 FL) and adult bone marrow (BM). More important, Lyl-1–/– E14 FL cells and BM are severely impaired in their competitive reconstituting abilities, especially with respect to B and T lineage reconstitution. Thus, ablation of Lyl-1 quantitatively and functionally affects HSCs, a cell population that transcribes Lyl-1 more actively than their differentiated progenies. Our results demonstrate for the first time that Lyl-1 functions are important for HSC properties and B-cell differentiation and that they are largely distinct from scl functions.
Chemokine receptor targeting efficiently directs antigens to MHC class I pathways and elicits antigen-specific CD8+ T-cell responses
Authors: Schiavo Roberta , Baatar Dolgor , Olkhanud Purevdorj , Indig Fred E. , Restifo Nicholas , Taub Dennis , Biragyn Arya
Published: March 2006

Chemokines are key controllers of cell trafficking and are involved in numerous pathologic and inflammatory conditions. However, the fate of a chemokine ligand, once it is endocytosed with its receptor, remains obscure. Here, using chemokine–tumor antigen fusion constructs, we demonstrate for the first time that chemokines are internalized to early/late endosomal and lysosomal compartments through a clathrin-dependent process and subsequently delivered to the cytosol for proteasomal processing, facilitating efficient cross-presentation to the TAP-1–dependent MHC class I processing pathway. These data not only elucidate the intracellular fate of chemokine ligands upon receptor uptake, but also demonstrate the superior carrier potency of chemokines for delivering self-antigens to both class I and II processing pathways to induce CD8+ and CD4+ T-cell responses.
Antibody-dependent cellular cytotoxicity (ADCC) is mediated by genetically modified antigen-specific human T lymphocytes
Authors: Clémenceau Béatrice , Congy-Jolivet Nicolas , Gallot Géraldine , Vivien Régine , Gaschet Joëlle , Thibault Gilles , Vié Henri
Published: March 2006

In the context of transplantation, donor and virus-specific T-lymphocyte infusions have demonstrated the dramatic potential of T cells as immune effectors. Unfortunately, most attempts to exploit the T-cell immune system against nonviral malignancies in the syngeneic setting have been disappointing. In contrast, treatments based on monoclonal antibodies (Abs) have been clinically successful and have demonstrated the clinical relevance of several antigens as therapeutic targets and the importance of the antibody-dependent cellular cytotoxicity (ADCC) pathway. In the present study, we considered the possibility of arming specific T cells with a receptor that would enable them to mediate ADCC. After transduction with a CD16/γ receptor gene, CD4+ and CD8+ cytotoxic T lymphocytes displayed stable expression of the CD16 receptor at their surface. In the absence of Ab, CD16/γ expression did not affect the capacity of specific T lymphocytes to kill their target following “natural” T-cell receptor recognition. When tested against the autologous B-lymphoblastoid cell line (BLCL) coated with anti-CD20 mAb, the newly expressed Fc receptor enabled the T cells to kill the BLCL through ADCC. Adoptive transfer of such newly designed immune effector may be considered to increase antibody efficiency by harnessing the immune potential of T cells.
New transgenic evidence for a system of sympathetic axons able to express tissue plasminogen activator (t-PA) within arterial/arteriolar walls
Authors: Hao Zhifang , Guo Caiying , Jiang Xi , Krueger Susan , Pietri Thomas , Dufour Sylvie , Cone Robert E. , O'Rourke James
Published: March 2006

Sympathetic axons embedded in a few arterioles and vasa vasora were recently shown to store tissue plasminogen activator (t-PA) in vesicles. But the extension of such t-PA axons to arteries and arterioles throughout the organism has not been verified. Confirmation of this anatomy would identify a second significant source of vessel wall t-PA. To visualize fine embedded axons independent of endothelium, we created a transgenic mouse whose expressions of the t-PA promoter and enhanced green fluorescent protein are confined to sympathetic neurons and other neural crest derivatives. Confocal images reveal the extension of t-PA axons to arterioles serving heart, brain, kidney, lung, mesentery, and skin; plus aortic, carotid, and mesenteric artery walls. Ganglion neurons and adrenal chromaffin cells also show strong expressions. These new sightings confirm the existence of a system of t-PA axons that is prominent in arterioles, and compatible with the release of neural t-PA into their walls. (Blood. 2006;108:200-202)
Effects of oral arsenic trioxide therapy on QT intervals in patients with acute promyelocytic leukemia: implications for long-term cardiac safety
Authors: Siu Chung-Wah , Au Wing-Yan , Yung Cindy , Kumana Cyrus R. , Lau Chu-Pak , Kwong Yok-Lam , Tse Hung-Fat
Published: March 2006

Ventricular tachyarrhythmias may occur during intravenous arsenic trioxide (As2O3). This has not happened during oral As2O3. Sixteen patients were studied by electrocardiography and 24-hour Holter monitoring at baseline, during and after oral As2O3 (As2O3-ON, As2O3-OFF). QT and corrected QT (QTc) were significantly longer during As2O3-ON than in As2O3-OFF, but QT and QTc dispersions were comparable. The patients' 24-hour heart rates were higher during As2O3-ON than in As2O3-OFF. QTc intervals at each hour were longer during As2O3-ON than in As2O3-OFF. However, QTc prolongation of more than 30 milliseconds only occurred at one time point (2 hours) after oral As2O3, resulting in QTc of more than 500 milliseconds in 3 of 16 patients, all within 4 hours of oral As2O3. Although the standard deviation of normal RR interval was lower during As2O3-ON, ratios of low frequency to high frequency power for As2O3-ON and As2O3-OFF were comparable. No ventricular proarrhythmias were observed. These observations, due to the lower peak plasma arsenic reached during oral As2O3, may explain the relative cardiac safety of oral As2O3. (Blood. 2006;108:103-106)
Bradykinin B2 receptor knockout mice are protected from thrombosis by increased nitric oxide and prostacyclin
Authors: Shariat-Madar Zia , Mahdi Fakhri , Warnock Mark , Homeister Jonathon W. , Srikanth Sujata , Krijanovski Yelena , Murphey Laine J. , Jaffa Ayad A. , Schmaier Alvin H.
Published: March 2006

Bradykinin (BK) liberates nitric oxide, prostacyclin, and tissue plasminogen activator from endothelial cells. We hypothesized that BK B2 receptor knockout (KO) mice (BKB2R-/-) have increased thrombosis risk. Paradoxically, the BKB2R-/- mice have long bleeding times and delayed carotid artery thrombosis, 78 ± 6.7 minutes, versus 31 ± 2.7 minutes in controls. The mechanism(s) for thrombosis protection was sought. In BKB2R-/- plasma coagulation, fibrinolysis and anticoagulant proteins are normal except for an increased prekallikrein and decreased factor XI. BKB2R-/- mice have elevated BK 1-5 (160 ± 75 fmol/mL, vs 44 ± 29 fmol/mL in controls) and angiotensin II (182 ± 41 pg/mL, vs 49 ± 7 pg/mL in controls). Ramipril treatment shortens vessel occlusion time. BKB2R-/- mice have elevated plasma 6-keto-PGF1α (666 ± 232 ng/mL, vs 23 ± 5.3 ng/mL in controls) and serum nitrate (61 ± 5.3 μM, vs 24 ± 1.8 μMin controls). Treatment with L-NAME (NG-mono-methyl-l-arginine ester) or nimesulide shortens the thrombosis time. BKB2R-/- mice have increased angiotensin receptor 2 (AT2R) mRNA and protein expression. Treatment with an AT2R antagonist, PD123 319, normalizes the thrombosis time and nitrate and 6-keto-PGF1α. The long bleeding times in BKB2R-/- mice also correct with L-NAME and nimesulide therapy. In BKB2R-/- mice, angiotensin II binding to an overexpressed AT2R promotes thromboprotection by elevating nitric oxide and prostacyclin. These investigations indicate a pathway for thrombosis risk reduction via the plasma kallikrein/kinin and renin angiotensin systems. (Blood. 2006;108:192-199)
Vascular endothelial cell–specific phosphotyrosine phosphatase (VE-PTP) activity is required for blood vessel development
Authors: Bäumer Sebastian , Keller Linda , Holtmann Astrid , Funke Ruth , August Benjamin , Gamp Alexander , Wolburg Hartwig , Wolburg-Buchholz Karen , Deutsch Urban , Vestweber Dietmar
Published: March 2006

VE-PTP, a receptor-type phosphotyrosine phosphatase, associates with the tyrosine kinase receptor Tie-2 and VE-cadherin and enhances the adhesive function of the latter. Here, VE-PTP was found to be restricted to endothelial cells, with a preference for arterial endothelium. Mutant mice expressing a truncated, secreted form of VE-PTP lacking the cytoplasmic and transmembrane domains and the most membrane-proximal extracellular fibronectin type III repeat, showed severe vascular malformations causing lethality at 10 days of gestation. Although blood vessels were initially formed, the intraembryonic vascular system soon deteriorated. Blood vessels in the yolk sac developed into dramatically enlarged cavities. In explant cultures of mutant allantoides, endothelial cells were found next to vessel structures growing as cell layers. No signs for enhanced endothelial apoptosis or proliferation were observed. Thus, the activity of VE-PTP is not required for the initial formation of blood vessels, yet it is essential for their maintenance and remodeling.
Transient accumulation of human mature thymocytes and regulatory T cells with CD28 superagonist in “human immune system” Rag2-/-γc-/- mice
Authors: Legrand Nicolas , Cupedo Tom , van Lent Anja U. , Ebeli Menno J. , Weijer Kees , Hanke Thomas , Spits Hergen
Published: March 2006

Efficient and quick reconstitution of T-cell compartments in lymphopenic patients is of great importance to prevent opportunistic infections, but remains difficult to achieve. Human T-cell proliferation in a T-cell-receptor (TCR)-independent manner is possible in vitro with superagonist anti-CD28 antibodies, and such molecules are therefore promising therapeutic tools. Here, we investigated the in vivo effects of superagonist anti-CD28 treatment on human developing and mature T cells, in the recently developed model of “human immune system” BALB/c Rag2-/-γc-/- mice. Our results show that superagonist anti-CD28 treatment transiently induces a 7-fold increase in thymocyte numbers and up to 18-fold accumulation of mature thymocytes. The increased thymic production lead to transient accumulation of mature T cells in the periphery at the peak of treatment effect (day 6). In addition, long-term peripheral T-cell depletion was induced. Furthermore, the concomitant selective expansion and accumulation of suppressive CD4+CD25+FoxP3+ T cells was induced in a transient manner. Superagonist anti-CD28 therapy could therefore be of clinical interest in humans, both for beneficial effect on thymic T-cell production as well as regulatory T-cell accumulation. (Blood. 2006;108:238-245)
Differential Spatial Approximation between Secretin and Its Receptor Residues in Active and Inactive Conformations Demonstrated by Photoaffinity Labeling
Authors: Dong Maoqing , Hosohata Keiko , Pinon Delia I. , Muthukumaraswamy Natesa , Miller Laurence J.
Published: March 2006

Understanding of the conformational changes in G protein-coupled receptors associated with activation and inactivation is of great interest. We previously used photoaffinity labeling to elucidate spatial approximations between photolabile residues situated throughout the pharmacophore of secretin agonist probes and this receptor. The aim of the current work was to develop analogous photolabile secretin antagonist probes and to explore their spatial approximations. The most potent secretin antagonist reported is a pseudopeptide ([ψ4, 5]secretin) in which the peptide bond between residues 4 and 5 was replaced by a ψ(CH2-NH) peptide bond isostere. We have developed a series of [ψ4, 5]secretin analogs incorporating photolabile benzoyl phenylalanine residues in positions 6, 22, and 26. Each bound to the secretin receptor saturably and specifically, with affinity similar to their parental peptide. At concentrations with no measurable agonist activity, each probe covalently labeled the secretin receptor. Peptide mapping using proteolytic cleavage, immunoprecipitation, and radiochemical sequencing identified that each of these three probes labeled the amino terminus of the secretin receptor. Whereas the position 22 probe labeled the same residue as its analogous agonist probe and the position 6 probe labeled a residue within two residues of that labeled by its analogous agonist probe, the position 26 probe labeled a site 16 residues away from that labeled by its analogous agonist probe. Thus, whereas structurally related agonist and antagonist probes dock in the same general region of this receptor, conformational differences in active and inactive states result in substantial differences in spatial approximation at the carboxyl-terminal end of secretin analogs.
Breath Analysis: Potential for Clinical Diagnosis and Exposure Assessment
Authors: Cao Wenqing , Duan Yixiang
Published: March 2006

Breath tests are among the least invasive methods available for clinical diagnosis, disease state monitoring, and environmental exposure assessment. In recent years, interest in breath analysis for clinical purposes has increased. This review is intended to describe the potential applications of breath tests, including clinical diagnosis of diseases and monitoring of environmental pollutant exposure, with emphasis on oxidative stress, lung diseases, metabolic disorder, gastroenteric diseases, and some other applications. The application of breath tests in assessment of exposure to volatile organic compounds is also addressed. Finally, both the advantages and limitations of breath analysis are summarized and discussed.
Dissociation of oocyte nuclear and cytoplasmic maturation by the addition of insulin in cultured bovine antral follicles
Authors: Fouladi-Nashta A A , Campbell K H S
Published: March 2006

Follicles of 4–8 mm in diameter were dissected from ovaries and cultured in Waymouth culture medium in the presence or absence of insulin (5 μg/ml) at 39 °C in a humidified atmosphere of 45% O2, 5% CO2 and 50% N2 for 24 h. Following follicle culture, the oocytes were collected and examined for developmental potential, total protein profile and ultrastructural aspects. Oocytes aspirated directly from follicles of the same size were used as controls. Addition of insulin to the follicle culture medium significantly reduced expression of the low molecular weight insulin-like growth factor-binding proteins (IGFBPs) in the follicular fluid, and significantly reduced the cleavage rate of subsequently matured and fertilised oocytes (0.52 vs 0.61). However, there were no differences in the proportion of cleaved embryos which developed to the blastocyst stage (0.30 vs 0.28), nor embryo quality as assessed by total cell number (137 ± 8.53 vs124.6 ± 6.95). The total protein profiles of immature oocytes recovered after 24 h of follicle culture were compared by PAGE. There were marked differences between the two groups, unmatured oocytes recovered from the insulin-positive follicle group showed a protein pattern similar to that of matured oocytes. In addition, examination of ultrastructural features by transmission electron microscopy indicated that oocytes from follicles cultured in the presence of insulin undergo many of the cytoplasmic changes associated with oocyte maturation. In conclusion, follicle culture in the presence of insulin is beneficial for follicular survival and significantly reduces cleavage but has no detrimental effects on the development of cultured embryos. However, many of the cytoplasmic changes associated with oocyte maturation occur prior to the induction of nuclear maturation.
The effects of IGF-I on bovine follicle development and IGFBP-2 expression are dose and stage dependent
Authors: Walters Kirsty A , Binnie John P , Campbell Bruce K , Armstrong David G , Telfer Evelyn E
Published: March 2006

This study aimed to determine the effect of insulin-like growth factor-I (IGF-I) on early antral bovine follicular development, and the expression of insulin-like growth factor-binding protein-2 (IGFBP-2). Antral follicles separated into three different size groups were cultured for 6 days in medium supplemented with either a low (10 ng/ml) or high (1 μg/ml) dose of human recombinant IGF-I. Oestradiol production by follicles in all size ranges, cultured in the presence of the high concentration of IGF-I, significantly increased by day 6 (P < 0.05). Follicles in the smallest size range, 165–215 μm, cultured in a high dose of IGF-I, were found to be significantly increased in size (P < 0.01). Oocyte health of the largest follicles (281–380 μm) was significantly improved by the addition of IGF-I to the culture medium. mRNA expression of IGFBP-2 was decreased in the granulosa cells of follicles, size range 216–280 μm, cultured with a high dose of IGF-I (P < 0.05). Granulosa cells (P < 0.05) and oocytes (P < 0.01) of the largest follicles (281–380 μm) showed a decrease in IGFBP-2 expression (protein) when cultured in the control and low-IGF-I treatment groups. Therefore, the response of a bovine follicle to IGF-I is both dose and stage dependent. This work supports a role for IGF-I in modulating somatic and germ-cell maturation and development in early antral follicles. Furthermore, the inverse relationship between the level of IGF-I stimulation and IGFBP-2 expression suggests a local regulatory system modulating IGF-I availability.
15-Hydroxyprostaglandin dehydrogenase in the bovine endometrium during the oestrous cycle and early pregnancy
Authors: Parent Marianne , Madore Eric , MacLaren Leslie A , Fortier Michel A
Published: March 2006

Prostaglandins (PG) are primary regulators of reproductive function. In ruminants, the relative production of PGE2 and PGF2α determines the return to a new oestrous cycle or to the establishment of pregnancy in response to a viable embryo. PG action depends on biosynthesis, transport and interaction with their receptors, which are all expressed differentially during the oestrous cycle. PGs are, however, local mediators and thus the onsite degradation by enzymes such as 15-hydroxyprostaglandin dehydrogenase (HPGD), also known as 15-PGDH, is another factor to consider in the regulation of physiological action. Little information is available on PG catabolism in the endometrium during the oestrous cycle or early pregnancy. The purpose of this study was to clone the bovine 15-PGDH, produce the recombinant protein and generate a specific antibody to study its activity and its expression in the endometrium during the oestrous cycle. We have found that the bovine 15-PGDH is highly homologous to the rat and human isoforms. 15-PGDH is localized principally in the glandular epithelium and to a lesser extent in stromal and luminal epithelial cells. The enzyme expression is regulated during the oestrous cycle and it reaches its maximal level on days 16–18. Transient expression is observed in luminal epithelial and trophoblast cells on day 21 of pregnancy. The mRNA is expressed at a constant high level throughout the cycle. The activity of the recombinant 15-PGDH was also tested and was found comparable for PGF2α and PGE2. These data suggest that 15-PGDH contributes to the tight regulation of PG action in the endometrium especially at the critical period of recognition of pregnancy.
N-cadherin mediated cell contact inhibits germinal vesicle breakdown in mouse oocytes maintained in vitro
Authors: Peluso J J
Published: March 2006

The effect of granulosa cell contact on the ability of zona-free oocytes to undergo germinal vesicle breakdown (GVBD) was assessed using a granulosa cell co-culture system. Oocytes contacted granulosa cells in a site-specific manner such that their GV was away from the granulosa cells. Also contact with granulosa cells reduced the percentage of oocytes undergoing GVBD from about 40% to 15%. GVBD was inhibited by contact with granulosa cells but not a granulosa cell-secreted product, since oocytes cultured in the same culture, that were adjacent to the granulosa cell monolayer underwent GVBD at the same rate as controls. Similarly, media collected from granulosa cell cultures did not attenuate the rate of GVBD. The ability of granulosa cell contact to inhibit GVBD was equal to that of db-cAMP. Moreover, the ability of granulosa cells to inhibit GVBD was not mimicked by spontaneously immortalized granulosa cells. This cell specificity appeared to be related to N-cadherin, since granulosa cells and oocytes express N-cadherin and a N-cadherin antibody attenuates the effect of granulosa cell contact. The mechanism through which N-cadherin mediated cell contact maintains meiotic arrest is unknown. It is possible that homophilic N-cadherin binding between the granulosa cells and oocyte acts through a junxtacrine mechanism, which in part may lead in the activation fibroblast growth factor (FGF) receptors that are expressed by the oocyte. The involvement of FGF receptors is supported by the observations that FGF and a N-cadherin peptide known to activate FGF receptors inhibit GVBD.
Identification of transcription factors at the site of implantation in the later stages of murine pregnancy
Authors: Zhao Baohui , Koon Deanna , Bethin Kathleen E
Published: March 2006

Despite medical advances, preterm delivery continues to complicate 12% of all births in the United States and is a major cause of neonatal deaths. One of the reasons that preterm labor continues to be a significant problem is that very little is understood about the factors involved in normal labor. Many investigators have studied parturition in the mouse and defined essential pathways for normal labor. Prostaglandins play an essential role in mouse labor and are important in human labor as well. We examined the 23 transcription factors from pregnant mouse uterus that change expression after the induction of cyclooxygenase-1, the enzyme that catalyzes the first committed step in prostaglandin synthesis. Using in situ hybridization, we have identified three of these transcription factors, Hoxa10, Hoxa11 and GILZ as being expressed in the decidua and regulated at the end of pregnancy. Both Hoxa10 and Hoxa11 are known to be critical for implantation, but very little is known about their roles in late gestation. GILZ has not previously been identified in the gravid uterus. In summary, we have identified three transcription factors that are regulated in the decidua at the end of pregnancy, suggesting a role in detachment of the fetus and placenta.
Activities of maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) are not required for the global histone deacetylation observed after germinal vesicle breakdown (GVBD) in porcine oocytes
Authors: Endo Tsutomu , Naito Kunihiko , Kume Sachi , Nishimura Yukio , Kashima Koji , Tojo Hideaki
Published: March 2006

The acetylation of nuclear core histone has been suggested to work as an epigenetic mark for transmitting gene expression patterns to daughter cells. Global histone deacetylations, presumably involved in the reprogramming of the gene expression, have been observed after germinal vesicle breakdown (GVBD) in a cell cycle-dependent manner during meiotic maturation of mouse and porcine oocytes, although the regulation mechanism of histone deacetylation has not been studied well. In the present study, we examined the involvement of a crucial cell-cycle-regulator, maturation-promoting factor (MPF), and a meiosis-related kinase, mitogen-activated protein kinase (MAPK), in the global histone deacetylation during porcine oocyte maturation. In order to know whether the activities of MPF and MAPK were required, or the breakdown of GV membrane was sufficient, for the global histone deacetylation observed after GVBD, we artificially destroyed the GV membrane of the porcine immature oocytes. The artificial GV destruction (AGVD) induced histone deacetylation without the activation of MPF and MAPK. This deacetylation after AGVD was not affected by an MPF inhibitor, roscovitine, or an inhibitor of protein synthesis, cycloheximide, but was completely prevented by an inhibitor of histone deactylases (HDACs), trichostatine A. HDAC1 was present in the GV of the immature oocytes and localized on chromosomes after GVBD and AGVD. These results suggest that the MPF and MAPK activities were dispensable and the breakdown of the GV membrane was sufficient for the global histone deacetylation, which was catalyzed by HDAC activity
Effect of intrafollicular indomethacin injection on gonadotropin surge-induced expression of select extracellular matrix degrading enzymes and their inhibitors in bovine preovulatory follicles
Authors: Li Qinglei , Jimenez-Krassel Fermin , Kobayashi Yasuhiro , Ireland James J , Smith George W
Published: March 2006

A growing body of evidence supports an obligatory role for intrafollicular prostanoids in the mechanism of ovulation. However, the prostanoid-dependent mediators of the follicular extracellular matrix degradation required for ovulation are unknown. The objectives of this study were to determine the cellular compartment(s) in which the gonadotropin surge-induced regulation of select extracellular matrix degrading enzymes and their cognate inhibitors occurs in bovine preovulatory follicles, and to test whether such regulation is blocked by intrafollicular administration of the prostanoid synthesis and ovulation inhibitor, indomethacin (INDO). Follicular fluid prostaglandin E2 concentrations were elevated in diluent-treated follicles before ovulation (24 h after GnRH injection), but the increase was blocked in INDO-treated follicles. Real-time PCR analysis revealed the specific follicular cell types where gonadotropin surge-induced increases in mRNA abundance for members of the matrix metalloproteinase/tissue inhibitor of metalloproteinase and plasminogen activator families occurred. INDO treatment increased thecal cell mRNA for tissue inhibitor of metalloproteinase-4 and its protein abundance in the apex of preovulatory follicles before ovulation, but suppressed granulosal cell mRNA and activity for tissue plasminogen activator in follicular fluid and the follicle apex. Plasmin activity was also suppressed in the follicular fluid of INDO-treated follicles. Effects of INDO injection on select matrix metalloproteinases were not observed. The results suggest that gonadotropin surge-induced regulation of tissue inhibitor of metalloproteinase-4 and tissue plasminogen activator may be prostanoid dependent, and support a potential role for increased tissue plasminogen activator expression and decreased tissue inhibitor of metalloproteinase-4 expression in the mechanism of ovulation.
Hormonal regulation and differential expression of neuropilin (NRP)-1 and NRP-2 genes in bovine granulosa cells
Authors: Shimizu Takashi , Jayawardana Barana C , Nishimoto Hiromi , Kaneko Etsushi , Tetsuka Masafumi , Miyamoto Akio
Published: March 2006

Although much is known about the biology of vascular endothelial growth factor (VEGF) and its receptors, little is known about the role of the VEGF receptors neuropilin (NRP)-1 and NRP-2 in the process of bovine follicle development. The aim of the present study was to examine the hormonal regulation of NRP-1 and NRP-2 mRNAs by real-time PCR in follicles from the bovine ovary and in cultured granulosa cells. The NRP-1 gene was expressed in the granulosa and theca cells in the post-selection (POF) and pre-selection (PRF) follicles in the bovine ovary. In contrast, the NRP-2 gene was expressed only in the theca cells in the POF and the PRF. The level of NRP-1 mRNA was significantly increased by treatment of the cultured granulosa cells with 10 ng/ml estradiol (E2). In contrast, the addition of progesterone (P4) to the culture medium decreased the expression of the NRP-1 gene. The level of NRP-1 mRNA was increased by 10 ng/ml E2 with or without 1 ng/ml P4, but the level of NRP-1 mRNA was decreased if the P4 level was increased to 10 ng/ml, even when 1 ng/ml E2 was also added. Follicle-stimulating hormone did not stimulate the expression of the NRP-1 gene. These results are the first data showing that NRP-1, but not NRP-2, is expressed in the granulosa cells of bovine follicles and that NRP-1 gene expression is regulated by sex steroids. Our findings suggest the involvement of NRP-1 in follicle development in the cow.
Role of the N, N′-dimethylbiguanide metformin in the treatment of female prepuberal BALB/c mice hyperandrogenized with dehydroepiandrosterone
Authors: Sander Valeria , Luchetti Carolina Griselda , Solano María Emilia , Elia Evelin , Di Girolamo Guillermo , Gonzalez Claudio , Motta Alicia Beatriz
Published: March 2006

The present study investigated the role of the N, N{′ }-dimethylbiguanide metformin (50 mg/100 g body weight in 0.05 ml water, given orally with a canulla) in the prevention of endocrine and immune disorders provoked by the hyperandrogenization with dehydroepiandrosterone (DHEA) in prepuberal BALB/c mice. The treatment with DHEA (6 mg/100 g body weight in 0.1 ml oil) for 20 consecutive days, recreates a mouse model that resembles some aspects of the human polycystic ovary syndrome (PCOS). The treatment with DHEA did not modify either body mass index (BMI) or blood glucose levels, but did increase fasting insulin levels when compared with controls. Markers of ovarian function – serum estradiol (E), progesterone (P) and ovarian prostaglandin E (PGE) – were evaluated. The treatment with DHEA increased serum E and P levels while ovarian PGE diminished. When metformin was administered together with DHEA, serum insulin, E and P levels, and ovarian PGE values did not differ when compared with controls. Using flow cytometry assays we found that the treatment with DHEA diminished the percentage of the CD4 + T lymphocyte population and increased the percentage of the CD8 + T lymphocyte population from both ovarian tissue and retroperitoneal lymph nodes. However, when metformin was administered together with DHEA, the percentages of CD4 + and CD8 + T lymphocyte populations from both ovarian tissue and retroperitoneal lymph nodes were similar to those observed in controls. Finally, when DHEA was administered alone it increased the serum tumor necrosis factor-alpha (TNF-α ) levels when compared with controls; however, when metformin was administered together with DHEA, serum TNF-α levels were similar to controls. These results indicate that metformin is able, directly or indirectly, to avoid the endocrine and immune alterations produced when mice are hyperandrogenized with DHEA.
Multiplex determination of murine seminal fluid cytokine profiles
Authors: Gopichandran Nadia , Ekbote Uma V , Walker James J , Brooke David , Orsi Nicolas M
Published: March 2006

Seminal fluid is known to be responsible for orchestrating mating-induced immunomodulation. Central to this process are numerous cytokines that modulate uterine leukocyte recruitment and trafficking. Despite this, a comprehensive analysis of the cytokine profile of murine seminal fluid is lacking. This study addressed this issue by using multiplex immunoassays to characterise the profile of interleukin (IL)-1α , IL-1β , IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-17, eotaxin, granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon (IFN)-γ, keratinocyte-derived chemokine (KC), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1α , MIP-1β , regulated upon activation normal T-cell expressed and secreted (RANTES), and tumour necrosis factor (TNF)-α in fluid drawn from the seminal vesicles of single mice (n = 18). Their levels and ratios were compared with those found in serum. IL-1α , IL-1β , IL-2, IL-5, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-17, GM-CSF, IFN-γ, MCP-1 and TNF-α levels were significantly higher in serum; IL-4, G-CSF, eotaxin, KC and RANTES exhibited the opposite trend. Based on these findings, we propose a model of mating-induced immunomodulation that implicates seminal eotaxin, RANTES and MIP-1α in the relocation and concentration of extravasated migrating endometrial eosinophils to the luminal epithelium. Furthermore, KC may participate in uterine neutrophil chemotaxis and activation. Eotaxin and MIP-α , together with IL-1β and IL-9, may also enhance further cytokine synthesis for endometrial antigen-presenting cell recruitment for processing paternal ejaculate antigens. IL-4 and G-CSF could also minimise deleterious cell-mediated immunity and modulate IFN-γ production, thereby supporting the establishment of pregnancy.
Morphologic changes in boar sperm nuclei with reduced disulfide bonds in electrostimulated porcine oocytes
Authors: Nakai Michiko , Kashiwazaki Naomi , Takizawa Akiko , Maedomari Naoki , Ozawa Manabu , Noguchi Junko , Kaneko Hiroyuki , Shino Masao , Kikuchi Kazuhiro
Published: March 2006

In pigs, failure of sperm nuclear decondensation has been reported after injection into oocytes. We examined the effects of pretreating sperm heads with Triton X-100 (TX-100) and dithiothreitol (DTT) and of electrical stimulation of oocytes after sperm head injection on time-dependent morphologic changes in sperm nuclei and in vitro development to the blastocyst stage. In experiment 1, spermatozoa were pretreated with 1% TX-100 and 5 mM DTT (T + D) or not treated, and then injected into in vitro matured oocytes. Electrical stimulation (1.5 kV/cm, 20 μs DC pulse) was applied to the oocytes 1 h after injection (stimulated group) or was not applied (unstimulated group). Some of the oocytes in each group were evaluated at hourly intervals until 10 h after injection for morphologic changes in the sperm nuclei. Unstimulated oocytes injected with untreated spermatozoa showed a delayed peak in the rate of nuclear decondensation (39.4–44.1%, 3–6 h after injection) compared with oocytes injected with T + D-treated spermatozoa (57.0% and 52.6%, 1 and 2 h, respectively). The rate of male pronucleus formation peaked 6 h after stimulation (by 40–60%) after injected oocytes had been stimulated with an electrical pulse, irrespective of whether or not the spermatozoa had been pretreated. In unstimulated oocytes, the rate of male pronucleus formation did not increase and stayed at the basal level (less than 20%) throughout the culture period, regardless of the sperm treatment. Thus, T + D treatment of spermatozoa did not affect completion of fertilization. In experiment 2, we evaluated the effects of electrical stimulation and sperm treatment with T + D on the rate of blastocyst formation and the mean number of cells per blastocyst. Oocytes stimulated after injection with either T + D-treated or untreated spermatozoa showed significantly higher percentages of blastocyst formation (24.8% and 27.1% respectively) than did unstimulated oocytes (1.1% and 4.1% for T + D-treated and untreated respectively; P < 0.01 by Duncan’s multiple-range test). The rate of blastocyst formation did not differ between the T + D-treated and untreated groups. The mean number of cells per blastocyst did not differ among any of the groups (14.0–29.4 cells). These results suggest that pretreatment of sperm with TX-100 and DTT shifted the timing of sperm nuclear decondensation forward. However, pronucleus formation and development to the blastocyst stage in vitro were not improved by sperm treatment. Thus, electrical stimulation of injected oocytes enhances in vitro development to the blastocyst stage in pigs.