Effect of preantral follicle isolation technique on in-vitro follicular growth, oocyte maturation and embryo development in mice

Background: The use of mechanical and enzymatic techniques to isolate preantral follicles before in-vitro culture has been previously described. The aim of this study was to assess the effect of the isolation procedure of mouse preantral follicles on their subsequent development in vitro. Methods: Follicles were isolated either mechanically or enzymatically and cultured using an individual non-spherical culture system. Follicular development and steroidogenesis, oocyte in-vitro maturation and embryo development were assessed for both groups. Results: After 12 days of culture, follicles isolated mechanically had a higher survival rate but a lower antral-like cavity formation rate than follicles isolated enzymatically. Enzymatic follicle isolation was associated with a higher production of testosterone and estradiol compared with mechanical isolation. A stronger phosphatase alkaline reaction was observed after enzymatic isolation, suggesting that follicles isolated enzymatically had more theca cells than those isolated mechanically. However, both isolation techniques resulted in similar oocyte maturation and embryo development rates. Conclusions: Enzymatic follicular isolation did not affect theca cell development. Follicular steroidogenesis was enhanced after enzymatic isolation but the developmental capacity of oocytes was comparable to that obtained after mechanical isolation.

Mechanobiology of the meniscus.

The meniscus plays a critical biomechanical role in the knee, providing load support, joint stability, and congruity. Importantly, growing evidence indicates that the mechanobiologic response of meniscal cells plays a critical role in the physiologic, pathologic, and repair responses of the meniscus. Here we review experimental and theoretical studies that have begun to directly measure the biomechanical effects of joint loading on the meniscus under physiologic and pathologic conditions, showing that the menisci are exposed to high contact stresses, resulting in a complex and nonuniform stress-strain environment within the tissue. By combining microscale measurements of the mechanical properties of meniscal cells and their pericellular and extracellular matrix regions, theoretical and experimental models indicate that the cells in the meniscus are exposed to a complex and inhomogeneous environment of stress, strain, fluid pressure, fluid flow, and a variety of physicochemical factors. Studies across a range of culture systems from isolated cells to tissues have revealed that the biological response of meniscal cells is directly influenced by physical factors, such as tension, compression, and hydrostatic pressure. In addition, these studies have provided new insights into the mechanotransduction mechanisms by which physical signals are converted into metabolic or pro/anti-inflammatory responses. Taken together, these in vivo and in vitro studies show that mechanical factors play an important role in the health, degeneration, and regeneration of the meniscus. A more thorough understanding of the mechanobiologic responses of the meniscus will hopefully lead to therapeutic approaches to prevent degeneration and enhance repair of the meniscus.

CD45+ Cells Present Within Mesenchymal Stem Cell Populations Affect Network Formation of Blood-Derived Endothelial Outgrowth Cells.

Mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs) represent promising cell sources for angiogenic therapies. There are, however, conflicting reports regarding the ability of MSCs to support network formation of endothelial cells. The goal of this study was to assess the ability of human bone marrow-derived MSCs to support network formation of endothelial outgrowth cells (EOCs) derived from umbilical cord blood EPCs. We hypothesized that upon in vitro coculture, MSCs and EOCs promote a microenvironment conducive for EOC network formation without the addition of angiogenic growth supplements. EOC networks formed by coculture with MSCs underwent regression and cell loss by day 10 with a near 4-fold and 2-fold reduction in branch points and mean segment length, respectively, in comparison with networks formed by coculture vascular smooth muscle cell (SMC) cocultures. EOC network regression in MSC cocultures was not caused by lack of vascular endothelial growth factor (VEGF)-A or changes in TGF-β1 or Ang-2 supernatant concentrations in comparison with SMC cocultures. Removal of CD45+ cells from MSCs improved EOC network formation through a 2-fold increase in total segment length and number of branch points in comparison to unsorted MSCs by day 6. These improvements, however, were not sustained by day 10. CD45 expression in MSC cocultures correlated with EOC network regression with a 5-fold increase between day 6 and day 10 of culture. The addition of supplemental growth factors VEGF, fibroblastic growth factor-2, EGF, hydrocortisone, insulin growth factor-1, ascorbic acid, and heparin to MSC cocultures promoted stable EOC network formation over 2 weeks in vitro, without affecting CD45 expression, as evidenced by a lack of significant differences in total segment length (p=0.96). These findings demonstrate the ability of MSCs to support EOC network formation correlates with removal of CD45+ cells and improves upon the addition of soluble growth factors.

The Influence Of Temperature And Salinity On Energy Partitioning In The Marine Nematode Diplolaimelloides-Bruciei

Measurements of population growth, generation time, fecundity and respiration in laboratory culture have been made, in relation to temperature and salinity, for the nematode Diplolaimelloides bruciei Hopper, a species normally associated with decayed material of the marsh grass Spartina. The intrinsic rate of increase (r) is high: it is related to temperature between 5° and 25°C by a sigmoid function which is steepest between 10° and 15°C, and is maximum at 26‰ salinity. Generation time is related to temperature by a power function and is shortest at 26‰ salinity. The effect of temperature on generation time is consistent with other data for marine nematodes, and the steep slope of r against temperature is largely due to the marked effect of temperature on fecundity. A sex ratio of 2:1 in favour of males is maintained regardless of culture conditions or population density. Respiration increases exponentially with temperature between 5° and 25°C, with a very high Q10 (3.94), but is not affected by salinity. At 30°C respiration is no higher than at 25°C. A high and relatively stable production efficiency (P/A) is maintained between 10 and 30°C with a maximum of 87% at 15°C; there is a stable reproductive effort (Pr/A) of about 10%. At 5°C both these ratios are zero. Data for the harpacticoid copepod Tachidius discipes, derived from the literature, show that this too has a high and stable production efficiency, which may be a characteristic of meiofaunal species in general, but in this species efficiency is relatively high at 5°C. Many features of the energy balance in D. bruciei can be related to an opportunistic mode of life.

Intercultural competence in the area of drug dependencies

This article takes a multidimensional or biopsychosocial conception of drug dependency as its starting point. Within this analytical framework, we advocate making the intercultural dimension more visible, since it is essential for the design and implementation of integral intervention processes. We propose intercultural competence as a working model that can increase the capacities of institutions and professionals —a particularly important consideration in the case of social work— in order to effectively address the aforementioned cultural dimension. After an extensive review of the scientific literature, we have defined five processes that can contribute to strengthening an institution’s intercultural competence and four processes that can do the same for a professional’s intercultural competence. Though selected for application in the area of drug dependencies, all these processes can also prove useful in improving attention to any other kind of culturally diverse group or person.

Calling for resistance: The political economy of Indian and Canadian call centre industries

Call centres have in the last three decades come to define the interaction between corporations, governments, and other institutions and their respective customers, citizens, and members. From telemarketing to tele-health services, to credit card assistance, and even emergency response systems, call centres function as a nexus mediating technologically enabled labour practices with the commodification of services. Because of the ubiquitous nature of the call centre in post-industrial capitalism, the banality of these interactions often overshadows the nature of work and labour in this now-global sector. Advances in telecommunication technologies and the globalization of management practices designed to oversee and maintain standardized labour processes have made call centre work an international phenomenon. Simultaneously, these developments have dislocated assumptions about the geographic and spatial seat of work in what is defined here as the new international division of knowledge labour. The offshoring and outsourcing of call centre employment, part of the larger information technology and information technology enabled services sectors, has become a growing practice amongst governments and corporations in their attempts at controlling costs. Leading offshore destinations for call centre work, such as Canada and India, emerged as prominent locations for call centre work for these reasons. While incredible advances in technology have permitted the use of distant and “offshore” labour forces, the grander reshaping of an international political economy of communications has allowed for the acceleration of these processes. New and established labour unions have responded to these changes in the global regimes of work by seeking to organize call centre workers. These efforts have been assisted by a range of forces, not least of which is the condition of work itself, but also attempts by global union federations to build a bridge between international unionism and local organizing campaigns in the Global South and Global North. Through an examination of trade union interventions in the call centre industries located in Canada and India, this dissertation contributes to research on post-industrial employment by using political economy as a juncture between development studies, critical communications, and labour studies.

Distinctive effects of G-CSF, GM-CSF and TNFalpha on neutrophil apoptosis in systemic lupus erythematosus.

OBJECTIVE: To investigate the influence of culture with G-CSF GM-CSF and TNFalpha on neutrophil apoptosis, comparing neutrophils from SLE patients with rheumatoid arthritis (RA) patients and healthy control subjects. METHODS: Neutrophils were isolated from SLE (n= 10), RA (n= 10) and healthy control subjects (n= 10), and cultured with two different concentrations of G-CSF, GM-CSF and TNFalpha. Proportion of apoptotic neutrophils at T=0, T=2hrs and T=24hrs was measured using FITC-labelled annexinV and flow cytometry. RESULTS: Significantly more neutrophils were apoptotic at T=0 in the SLE subjects than in the other groups (median, range--Control 3.5% (0.3-7.9) SLE 9.5% (2.9-29.1) RA 3.0% (0.4-23.0) p

Bioreactor Expansion of Human Adult Bone Marrow-Derived Mesenchymal Stem Cells

Supplementation of mesenchymal stem cells (MSCs) during hematopoietic stem cell transplantation (HSCT) alleviates complications such as graft-versus-host disease, leading to a speedy recovery of hematopoiesis. To meet such clinical demand, a fast MSCs expansion method is required. In the present study, we examined the feasibility of expanding MSCs from the isolated bone marrow mononuclear cells using a rotary bioreactor system. The cells were cultured in a rotary bioreactor with Myelocult� medium containing a combination of supplementary factors, including stem cell factor (SCF), interleukin 3 and 6 (IL-3, IL-6). After 8 days of culture, total cell numbers, Stro-1+CD44+CD34- MSCs and CD34+CD44+Stro-1- HSCs were increased 9, 29, and 8 folds respectively. Colony forming efficiency-fibroblast per day (CFE-F/day) of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. The bioreactor-expanded MSCs showed expression of primitive MSCs markers endoglin (SH2) and vimentin, whereas markers associated with lineage differentiation including osteocalcin (osteogenesis), Type II collagen (chondrogenesis) and C/EBPα (adipogenesis) were not detected. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes and adipocytes. Taken together, we conclude that the rotary bioreactor with the modified Myelocult� medium reported in this study may be used to rapidly expand MSCs.

Cultural policy models and cultural facilities models: from national to local models. Analysis of Barcelona’s case

The development of cultural policy analysis by social science has been produced a theorization about cultural policy models from sociology and political science. This analysis shows the influence of the national model of cultural policy on the forms of governance and management of cultural facilities. However, in this paper we will defend that currently the local model of cultural policy decisively influences the model of cultural institutions. This is explained by the growing importance of culture in local development strategies. In order to demonstrate this we will analyze the case of the Barcelona Model of local development and cultural policy, that is characterized for the level of local government leadership, multilevel governance, the use of culture in urban planning processes and a tendency to use public-private partnership in public management. This Model influences the genesis and development of the cultural facilities and it produces a singular and relatively successful model.

Cultural assets and sustainable development: the economic significance of Cultural Heritage

This article assesses the condition of the Cultural Heritage as a form of capital that gives rise to a significant flow of economic returns widely outweighing the effort it takes to preserve it. More specifically, the data related to Spain is provided from the perspective of aggregate demand drawing up an estimation of both the direct and indirect economic impacts arising from the Cultural Heritage valuation. The results highlight again the relevance of cultural tourism in the delivery of these economic returns and as a catalyst of activities leading to the sustainable socioeconomic devel-opment of multiple territories.

Confusing Culture and Politics: Ulster Scots Culture and Music

In Northern Ireland of the mid-1990s, many were caught off-guard by the comet-like arrival of a previously unarticulated Ulster Scots identity, replete with its own folk traditions. The article exploits the author’s position as Traditional Arts Officer of the Arts Council of Northern Ireland from 1998 until 2003 to offer an auto-ethnographic reflection on the literature on cultural politics in Northern Ireland since the outbreak of the Troubles, informed by extensive fieldwork with arts administrators and with contemporary Ulster Scots musicians. The article gives a close reading and historical contextualisation of the Ulster-Scots musical revue On Eagle's Wing, which was developed during this period. The article uses the concept of "suture" articulated by Lacan and developed by Laclau, Mouffe, and Žižek to explain both the potential for Ulster Scots culture to enter political discourse out of a seeming vacuum and its subsequent difficulties, offering an original interpretation of the role of culture in the contemporary trajectory of Northern Ireland politics.

Characterization of a spontaneous mouse retinal pigment epithelial cell line B6-RPE07.

PURPOSE. A spontaneously arising retinal pigment epithelial (RPE) cell line (B6-RPE07) was cloned from a primary culture of mouse RPE cells and maintained in culture for more than 18 months. Morphologic and functional properties of this cell line have been characterized.

METHODS. The morphology of the B6-RPE07 cells was examined by phase-contrast light microscopy, electron microscopy, and confocal microscopy. Barrier properties were measured by the flux of fluorescence from the apical to the basolateral compartment of culture chambers. The abilities of the cells to bind/phagocytose photoreceptor outer segments (POS) were determined by confocal microscopy, electron microscopy, and flow cytometry. Cytokine/chemokine secretion was measured by cytometric bead array. The expression of visual cycle proteins was determined by RT-PCR and Western blotting.

RESULTS. In standard culture conditions, B6-RPE07 cells display cobblestone morphology. When cultured on three-dimensional (3D) collagen gel–coated membranes, B6-RPE07 cells exhibit a monolayer epithelial polarization with apical surface microvilli. Immunohistochemistry of B6-RPE07 cultures revealed a high expression of pan-cytokeratin. B6-RPE07 cells also expressed the retinal pigment epithelium-specific marker CRALBP, but not RPE65. Cell junction proteins ZO-1 and ß-catenin, but not claudin-1/3 or occludin-1, were observed in B6-RPE07 cells. B6-RPE07 cells are able to bind, phagocytose, and digest POS. Finally, B6-RPE07 cells produce high levels of IL-6 and CCL2.

CONCLUSIONS. This is the first report of a mouse RPE cell line with morphology, phenotype, and function similar to those of in vivo mouse RPE cells. This cell line will be a valuable resource for future RPE studies, in particular for in vivo gene modification and transplantation studies.

From Empire to Iraq and the "War on Terror": The Transplanation and Commodification of the (Northern) Irish Policing Experience

Charting the enduring export appeal of policing models from (Northern) Ireland, this article sheds some light on the processes by which policing models are communicated and actively promoted to the global policing environment. The authors demonstrate how the transplantation of the Irish colonial model (ICM) represents an early example of the globalization of policing. The legacy of counterinsurgency expertise embedded within the ICM remains a historical constant and is a key factor in relation to the increasing commodification of the contemporary Northern Irish policing model, a model that successfully blends counterterrorism experience with a template for democratic policing reform. By juxtaposing these models, the authors provide a conceptual framework through which to assess the contemporary substance of policing transfer. The authors conclude by suggesting that the seductiveness of these policing models is largely attributable to lessons in counterinsurgency and notions of "Ireland as the solution" to a host of complex security scenarios.

The Challenges of Universalising Religions: the Kabbalah Centre in France and Britain.

The Kabbalah Centre is an offshoot of Judaism which since the 1990s has spread kabbalistic teaching in several countries to a religiously diverse audience. This article compares two European branches of the Kabbalah Centre: the flourishing London Centre, and the Parisian Centre that declined in the late 1990s before closing its doors in 2005. It emphasizes in particular the responses they stirred from the media, anticult movements, Orthodox Judaism and the Jewish population. Ultimately, this case study allows us to observe, in situ, the trajectory of a global religion, torn between its Jewish roots and universalistic ambitions. It emphasizes the importance, in this process, of the relationship it maintains, willingly or not, with its original religious frame. Consequently, the importance of local contexts is raised, illustrating the impact and combination of diverse factors. In addition to public and official responses to religious diversity, religious movements are affected by the religious landscape and the structures and authorities of religious organizations, as well as the religious and cultural characteristics of the population. Ultimately, this article underscores the complexity of the globalization of religion, which embraces a wide range of complex, sometimes ambiguous, situations lying between strong particularistic identity-claims and cosmopolitan, universalistic ambitions.