The Extracellular Environment\u27s Effect on Cellular Processes: An In Vitro Study of Mechanical and Chemical Cues on Human Mesenchymal Stem Cells and C17.2 Neural Stem Cells

Stem cells are widely used in the area of tissue engineering. The ability of cells to interact with materials on the nano- and micro- level is important in the success of the biomaterial. It is well-known that cells respond to their micro- and nano-environments through a process termed chemo-mechanotransduction. It is important to establish standard protocols for cellular experiments, as chemical modifications to maintenance environments can alter long-term research results. In this work, the effects of different media compositions on human mesenchymal stem cells (hMSCs) throughout normal in vitro maintenance are investigated. Changes in RNA regulation, protein expression and proliferation are studied via quantitative polymerase chain reaction (qPCR), immunocytochemistry (ICC) and cell counts, respectively. Morphological differences are also observed throughout the experiment. Results of this study illustrate the dynamic response of hMSC maintenance to differences in growth medium and passage number. These experiments highlight the effect growth medium has on in vitro experiments and the need of consistent protocols in hMSC research. A substantial opportunity exists in neuronal research to develop a material platform that allows for both the proliferation and differentiation of stem cells into neurons and the ability to quantify the secretome of neuronal cells. Anodic aluminum oxide (AAO) membranes are fabricated in a two-step anodization procedure where voltage is varied to control the pore size and morphology of the membranes. C17.2 neural stem cells are differentiated on the membranes via serum-withdrawal. Cellular growth is characterized by scanning electron microscopy (SEM), ICC and qPCR. ImageJ software is used to obtain phenotypic cell counts and neurite outgrowth lengths. Results indicate a highly tunable correlation between AAO nanopore sizes and differentiated cell populations. By selecting AAO membranes with specific pore size ranges, control of neuronal network density and neurite outgrowth length is achievable. To understand differentiation marker expressions in C17.2 NSCs and how material stiffness affects differentiation, cells are cultured on substrates of varying stiffness. qPCR is used to analyze neural stem cell, neural progenitor cell, neuron-restricted progenitor and differentiated post-mitotic neuronal cell RNA expression. Results suggest a relationship between material stiffness and neuronal development in C17.2 neural stem cells

Medium development strategies and scale down models for a high density high productivity cell line

Medium Development at Regeneron continues to enhance fed batch culture productivity. These efforts have been enabled through the development of high throughput scale down models in shake flasks and the ambr® 250. Design of Experiment (DOE) approaches have been applied to optimize the operating conditions in the small scale models leading to performance for growth and titer that match benchtop bioreactor with no off-set. The development of these representative scale down models and our approach to medium development will be described. A medium development case study will be presented from a recent Regeneron fed batch process with a cell line achieving high cell densities and depleting the culture of key amino acids. The traditional medium development approach of supplementing the culture with the depleted nutrients was unsuccessful: high amino acid consumption rates required large amounts of amino acids resulting in significantly increased culture osmolality and reduced productivity. Leveraging high throughput culture systems and multifactor DOEs, multiple medium composition factors in combination were rapidly evaluated. Mathematical models relating medium input factors to process outputs are generated that allow for process optimization. Using this approach, a new feeding strategy was developed that limits increases in osmolality and yields titers approaching 10g/L in both the scale down systems and a process that has been implemented for clinical scale manufacturing of a monoclonal antibody

Progress and gaps in reproductive health services in three humanitarian settings: mixed-methods case studies

Background Reproductive health (RH) care is an essential component of humanitarian response. Women and girls living in humanitarian settings often face high maternal mortality and are vulnerable to unwanted pregnancy, unsafe abortion, and sexual violence. This study explored the availability and quality of, and access barriers to RH services in three humanitarian settings in Burkina Faso, Democratic Republic of the Congo (DRC), and South Sudan. Methods Data collection was conducted between July and October 2013. In total, 63 purposively selected health facilities were assessed: 28 in Burkina Faso, 25 in DRC, and nine in South Sudan, and 42 providers completed a questionnaire to assess RH knowledge and attitudes. Thirty-four focus group discussions were conducted with 29 members of the host communities and 273 displaced married and unmarried women and men to understand access barriers. Results All facilities reported providing some RH services in the prior three months. Five health facilities in Burkina Faso, six in DRC, and none in South Sudan met the criteria as a family planning service delivery point. Two health facilities in Burkina Faso, one in DRC, and two in South Sudan met the criteria as an emergency obstetric and newborn care service delivery point. Across settings, three facilities in DRC adequately provided selected elements of clinical management of rape. Safe abortion was unavailable. Many providers lacked essential knowledge and skills. Focus groups revealed limited knowledge of available RH services and socio-cultural barriers to accessing them, although participants reported a remarkable increase in use of facility-based delivery services. Conclusion Although RH services are being provided, the availability of good quality RH services was inconsistent across settings. Commodity management and security must be prioritized to ensure consistent availability of essential supplies. It is critical to improve the attitudes, managerial and technical capacity of providers to ensure that RH services are delivered respectfully and efficiently. In addition to ensuring systematic implementation of good quality RH services, humanitarian health actors should meaningfully engage crisis-affected communities in RH programming to increase understanding and use of this life-saving care

Development of scale down models for perfusion bioreactor medium optimization

Due to the complex nature of balancing \u3e50 individual media components, the development and optimization of bioreactor medium for high performing perfusion bioreactors is a resource intensive, multivariate problem that greatly benefits from the availability of predictive high through-put scale-down models that simulate the bioreactor system. For that purpose, both a 10 mL long-term block model and 50 mL shaker tube model were developed and optimized to settings that balance oxygen transfer, culture health, and productivity. The long-term block model was limited by the volume needed for culture sampling; as a result, the shaker tube model was developed with a 7.5x increase in working volume. This shaker tube model was then applied to adequately characterize cell nutrient consumption profiles and subsequently inform medium development through multivariate design of experiments (DOE). Within two rounds of studies in the scale-down models, Regeneron’s first-generation perfusion medium formulation achieved approximately 100% increase in productivity compared to the initial medium. The improved nutrient strategy optimized in shaker tubes translated to several cell lines in the benchtop and pilot scale bioreactor perfusion system, indicating the predictive capabilities of the small-scale model. These results highlight the benefits of using small-scale models to shorten development time for perfusion process implementation

Etiology of Teen Dating Violence among Adolescent Children of Alcoholics

Family processes in early life have been impli- cated in adolescent involvement in teen dating violence, yet the developmental pathways through which this occurs are not well understood. In this study, etiological pathways from parental psychopathology and marital conflict in infancy to involvement in dating violence in late adoles- cence were examined in a sample of children at high-risk due to parental alcohol problems. Families (N = 227) recruited when the child was 12 months of age were assessed at 12-, 24-, 36-months, kindergarten, 6th, 8th, and 12th grades. Slightly more than half of the children were female (51%) and the majority were of European American descent (91%). Parental psychopathology in infancy was indirectly associated with teen dating violence in late adolescence via low maternal warmth and self-regulation in early childhood, externalizing behavior from kindergarten to early adolescence, and sibling problems in middle childhood. Marital conflict was also indirectly associated with teen dating violence via child externalizing behavior. Maternal warmth and sensitivity in early childhood emerged as an important protective factor and was associated with reduced marital conflict and increased child self-regulation in the preschool years as well as increased parental monitoring in middle childhood and early adolescence. Family processes occurring in the preschool years and in middle childhood appear to be critical periods for creating condi- tions that contribute to dating violence risk in late adoles- cence. These findings underscore the need for early intervention and prevention with at-risk families

Meeting the demand of women affected by ongoing crisis: Increasing contraceptive prevalence in North and South Kivu, Democratic Republic of the Congo

Context Over 20 years of conflict in the DRC, North and South Kivu have experienced cycles of stability and conflict, resulting in a compromised health system and poor sexual and reproductive health outcomes. Modern contraceptive use is low (7.5%) and maternal mortality is high (846 deaths per 100,000 live births). Program partners have supported the Ministry of Health (MOH) in North and South Kivu to provide good quality contraceptive services in public health facilities since 2011. Methods Cross-sectional population-based surveys were conducted in the program areas using a two-stage cluster sampling design to ensure representation in each of six rural health zones. Using MOH population estimates for villages in the catchment areas of supported health facilities, 25 clusters in each zone were selected using probability proportional to size. Within each cluster, 22 households were systematically selected, and one woman of reproductive age (15–49 years) was randomly selected from all eligible women in each household. Results Modern contraceptive prevalence among women in union ranged from 8.4% to 26.7% in the six health zones; current use of long-acting or permanent method (LAPM) ranged from 2.5% to 19.8%. The majority of women (58.9% to 90.2%) reported receiving their current method for the first time at a health facility supported by the program partners. Over half of women in four health zones reported wanting to continue their method for five years or longer. Conclusion Current modern contraceptive use and LAPM use were high in these six health zones compared to DRC Demographic and Health Survey data nationally and provincially. These results were accomplished across all six health zones despite their varied socio-demographic characteristics and different experiences of conflict and displacement. These findings demonstrate that women in these conflict-affected areas want contraception and will choose to use it when good quality services are available to them

“Provide care for everyone please”: Engaging Community Leaders as Sexual and Reproductive Health Advocates in North and South Kivu, Democratic Republic of Congo

Background: Inadequate infrastructure, security threats from ongoing armed conflict, and conservative socio-cultural and gender norms that favour large families and patriarchal power structures contribute to poor sexual and reproductive health (SRH) outcomes in North and South Kivu provinces, Democratic Republic of the Congo (DRC). In order to expand contraceptive and post-abortion care (PAC) access in North and South Kivu, CARE, the International Rescue Committee and Save the Children provided technical support to the Ministry of Health and health facilities in these regions. Partners acknowledged that community leaders, given their power to influence local customs, could play a critical role as agents of change in addressing inequitable gender norms, stigma surrounding SRH service utilization, and topics traditionally considered taboo within Congolese society. As such, partners actively engaged with community leaders through a variety of activities such as community mapping exercises, values clarification and transformation (VCAT) activities, situational analyses, and education. Methods: This manuscript presents findings from 12 key informant interviews (KIIs) with male political and non-political community leaders conducted in six rural health zones of North and South Kivu, DRC. Transcripts were analysed thematically to explore community leaders’ perceptions of their role in addressing the issue of unintended pregnancy in their communities. Results: While community leaders in this study expressed overall positive impressions of contraception and strong support for ensuring access to PAC services following spontaneous and induced abortions, the vast majority held negative beliefs concerning women who had induced abortion. Contrasting with their professed opposition to induced abortion, leaders’ commitment to mediating interpersonal conflict arising between community members and women who had abortions was overwhelming. Conclusion: Results from this study suggest that when thoughtfully engaged by health interventions, community leaders can be empowered to become advocates for SRH. While study participants were strong supporters of contraception and PAC, they expressed negative perceptions of induced abortion. Given the hypothesized link between the presence of induced abortion stigma and care-avoidance behavior, further engagement and values clarification exercises with leaders must be integrated into community mobilization and engagement activities in order to increase PAC utilization

Contraceptive use among adolescent and young women in North and South Kivu, Democratic Republic of the Congo: A cross-sectional population-based survey

Background: Adolescent girls in humanitarian settings are especially vulnerable as their support systems are often disrupted. More than 20 years of violence in the Democratic Republic of the Congo (DRC) has weakened the health system, resulting in poor sexual and reproductive health (SRH) outcomes for women. Little evidence on adolescent contraceptive use in humanitarian settings is available. CARE, International Rescue Committee (IRC), and Save the Children, in collaboration with the Reproductive Health Access, Information and Services in Emergencies (RAISE) Initiative, Columbia University, have supported the Ministry of Health (MOH) since 2011 to provide good quality contraceptive services in public health facilities in conflict-affected North and South Kivu. In this study, we analyzed contraceptive use among sexually active young women aged 15-–24 in the health zones served by the partners’ programs. Methods and findings: The partners conducted cross-sectional population-based surveys in program areas of North and South Kivu using two-stage cluster sampling in six health zones in July-–August 2016 and 2017. Twenty-five clusters were selected in each health zone, 22 households in each cluster, and one woman of reproductive age (15–-49 years) was randomly selected in each household. This manuscript presents results from a secondary data analysis for 1,022 women aged 15–-24 who reported ever having sex: 326 adolescents (15–-19 years) and 696 young women (20–-24 years olds ), 31.7% (95% confidence interval [(CI]) 29.5–-34.1), of whom were displaced at least once in the previous five years. Contraceptive knowledge was high, with over 90% of both groups able to name at least one modern contraceptive method. Despite this high knowledge, unmet need for contraception was also high: 31.7% (95%CI 27.9–-35.7) among 15–-19- year- olds and 40.1% (95% CI 37.1–-43.1, p = 0.001) among 20–-24- year- olds. Current modern contraceptive use (16.5%, 95% CI, 14.7–-18.4) was similar in both age groups, the majority of whom received their method from a supported health facility. Among current users, more than half of 15–-19- year- olds were using a long-acting reversible contraceptive (LARC; 51.7%, 95% CI 41.1–-61.9) compared to 36.5% of 20–-24- year- olds (95% CI 29.6–-43.9, p = 0.02). Age, Being unmarried, younger age of sexual debut, having some secondary education, being unmarried, and having begun childbearing were associated with modern contraceptive use. The main limitations of our study are related to insecurity in three health zones that prevented access to some villages, reducing the representativeness of our data, and our defining sexually active women as those who have ever had sex. Conclusions: In this study, to our knowledge one of the first to measure contraceptive prevalence among adolescents in a humanitarian setting, we observed that adolescent and young women will use modern contraception, including long-acting methods. Meaningful engagement of adolescent and young women would likely contribute to even better outcomes. Creating an enabling environment by addressing gender and social norms, however, are is key to reducing stigma and meeting the demand for contraception of young women. As we continue to build such supportive environments, we can see that they will use effective contraception when contraceptive services, including short- and long-acting methods, are available, even in protracted crisis settings

Optically-passive spirals: The missing link in gradual star formation suppression upon cluster infall

Galaxies migrate from the blue cloud to the red sequence when their star formation is quenched. Here, we report on galaxies quenched by environmental effects and not by mergers or strong AGN as often invoked: They form stars at a reduced rate which is optically even less conspicuous, and manifest a transition population of blue spirals evolving into S0 galaxies. These 'optically passive' or 'red spirals' are found in large numbers in the STAGES project (and by Galaxy Zoo) in the infall region of clusters and groups.Comment: Proceedings of "The Starburst-AGN connection" conference held in Shanghai, Oct 27-31, 200

Obscured star formation in intermediate-density environments:A Spitzer study of the Abell 901/902 supercluster

We explore the amount of obscured star formation as a function of environment in the Abell 901/902 (A901/902) supercluster at z = 0.165 in conjunction with a field sample drawn from the A901 and CDFS fields, imaged with the Hubble Space Telescope as part of the Space Telescope A901/902 Galaxy Evolution Survey and Galaxy Evolution from Morphology and Spectral Energy Distributions (SEDs) Survey. We combine the combo-17 near-UV/optical SED with Spitzer 24 mu m photometry to estimate both the unobscured and obscured star formation in galaxies with M-* > 10(10) M-circle dot. We find that the star formation activity in massive galaxies is suppressed in dense environments, in agreement with previous studies. Yet, nearly 40% of the star-forming (SF) galaxies have red optical colors at intermediate and high densities. These red systems are not starbursting; they have star formation rates (SFRs) per unit stellar mass similar to or lower than blue SF galaxies. More than half of the red SF galaxies have low infrared-to-ultraviolet (IR-to-UV) luminosity ratios, relatively high Sersicindices, and they are equally abundant at all densities. They might be gradually quenching their star formation, possibly but not necessarily under the influence of gas-removing environmental processes. The other greater than or similar to 40% of the red SF galaxies have high IR-to-UV luminosity ratios, indicative of high dust obscuration. They have relatively high specific SFRs and are more abundant at intermediate densities. Our results indicate that while there is an overall suppression in the SF galaxy fraction with density, the small amount of star formation surviving the cluster environment is to a large extent obscured, suggesting that environmental interactions trigger a phase of obscured star formation, before complete quenching