Immunological assays for chemokine detection in in-vitro culture of CNS cells

Herein we review the various methods currently in use for determining the expression of chemokines by CNS cells in vitro. Chemokine detection assays are used in conjuction with one another to provide a comprehensive, biologically relevant assessment of the chemokines which is necessary for correct data interpretation of a specific observed biological effect. The methods described include bioassays for soluble chemokine receptors, RNA extraction, RT-PCR, Real - time quantitative PCR, gene array analysis, northern blot analysis, Ribonuclease Protection assay, Flow cytometry, ELISPOT, western blot analysis, and ELISA. No single method of analysis meets the criteria for a comprehensive, biologically relevant assessment of the chemokines, therefore more than one assay might be necessary for correct data interpretation, a choice that is based on development of a scientific rationale for the method with emphasis on the reliability and relevance of the method

Summary findings from a mixed methods study on identifying and responding to maternal and newborn illness in seven countries: implications for programs

Background: There is a lack of systematic information documenting recognition of potentially life-threatening complications and decisions to seek care, as well as reaching care and the specific steps in that process. In response to this gap in knowledge, a multi-country mixed methods study was conducted to illuminate the dynamics driving Delays 1 and 2 across seven countries for maternal and newborn illness and death. Methods: A common protocol and tools were developed, adapted by each of seven study teams depending on their local context (Ethiopia, India, Indonesia, Nigeria, Tanzania, Uganda, and Nepal). Maternal and newborn illness, and maternal and newborn death cases were included. Trained interviewers conducted event narratives to elicit and document a detailed sequence of actions, from onset of symptoms to the resolution of the problem. Event timelines were constructed, and in-depth interviews with key informants in the community were conducted. Transcripts were coded and analyzed for common themes corresponding to the three main domains of recognition, decision-making, and care-seeking. Results: Maternal symptom recognition and decision-making to seek care is faster than for newborns. Perceived cause of the illness (supernatural vs. biological) influences the type of care sought (spiritual/traditional vs. formal sector, skilled). Mothers, fathers, and other relatives tend to be the decision-makers for newborns while husbands and elder females make decisions for maternal cases. Cultural norms such as confinement periods and perceptions of newborn vulnerability result in care being brought in to the home. Perceived and actual poor quality of care was repeatedly experienced by families seeking care. Conclusion: The findings link to three action points: (1) messaging around newborn illness needs to reinforce a sense of urgency and the need for skilled care regardless of perceived cause; (2) targeted awareness building around specific maternal danger signs that are not currently recognized and where quality care is available is needed; and (3) designing appropriate contextualized messages. This research links to and supports a number of current global initiatives such as Ending Preventable Maternal Mortality, the Every Newborn Action Plan, the WHO Quality of Care framework, and the WHO guidelines on simplified management of newborn sepsis at the community level. This type of research is invaluable for designing programs to improve maternal and newborn survival to achieve ambitious global targets

Effects of User Fee Exemptions on the Provision and Use of Maternal Health Services: A Review of Literature

User fee removal has been put forward as an approach to increasing priority health service utilization, reducing impoverishment, and ultimately reducing maternal and neonatal mortality. However, user fees are a source of facility revenue in many low-income countries, often used for purchasing drugs and supplies and paying incentives to health workers. This paper reviews evidence on the effects of user fee exemptions on maternal health service utilization, service provision, and outcomes, including both supply-side and demand-side effects. We reviewed 19 peer-reviewed research articles addressing user fee exemptions and maternal health services or outcomes published since 1990. Studies were identified through a USAIDcommissioned call for evidence, key word search, and screening process. Teams of reviewers assigned criteria- based quality scores to each paper and prepared structured narrative reviews. The grade of the evidence was found to be relatively weak, mainly from short-term, non-controlled studies. The introduction of user fee exemptions appears to have resulted in increased rates of facility-based deliveries and caesarean sections in some contexts. Impacts on maternal and neonatal mortality have not been conclusively demonstrated; exemptions for delivery care may contribute to modest reductions in institutional maternal mortality but the evidence is very weak. User fee exemptions were found to have negative, neutral, or inconclusive effects on availability of inputs, provider motivation, and quality of services. The extent to which user fee revenue lost by facilities is replaced can directly affect service provision and may have unintended consequences for provider motivation. Few studies have looked at the equity effects of fee removal, despite clear evidence that fees disproportionately burden the poor. This review highlights potential and documented benefits (increased use of maternity services) as well as risks (decreased provider motivation and quality) of user fee exemption policies for maternal health services. Governments should link user fee exemption policies with the replacement of lost revenue for facilities as well as broader health system improvements, including facility upgrades, ensured supply of needed inputs, and improved human resources for health. Removing user fees may increase uptake but will not reduce mortality proportionally if the quality of facility-based care is poor. More rigorous evaluations of both demand- and supply-side effects of mature fee exemption programmes are needed

Effector cell mediated cytotoxicity measured by intracellular Granzyme B release in HIV infected subjects

CD8+ cytotoxic T lymphocyte (CTL) activity is currently believed to be one of the key immunologic mechanisms responsible for the prevention or attenuation of HIV-1 infection. The induction of CD8+ T cell activation may also result in the production of soluble or non-classical lytic factors that are associated with protection from infection or slower disease progression. Traditionally, CD8+ CTL responses have been measured by the classic chromium release assay, monitoring the ability of T cells (Effector cells) to lyse radiolabelled HLA – matched “target cells” that express the appropriate antigen-MHC complex. This method is not only labor intensive, semi quantitative assay at best, but also needs fresh, non-cryopreserved cells. Recently, cytokine specific ELISPOT assays or tetrameric MHC-I/ peptide complexes have utilized to directly quantitate circulating CD8+ effector cells, and these assays are more sensitive, quantitative and reproducible than the traditional CTL lysis assay and can also be performed on cryopreserved cells. Although these are reproducible assays for the assessment of soluble antiviral activity secreted by activated T cell populations they can be extremely expensive to perform. We have used FACS Analysis to measure Granzyme B release as a function of cell mediated cytotoxicity. This method helps quantitate the CTL activity and also identifies the phenotype of the cells elucidating this immune response. The method described not only monitors immunological response but also is also simple to perform, precise and extremely time efficient and is ideal for screening a large number of samples

Methodology for a mixed-methods multi-country study to assess recognition of and response to maternal and newborn illness

Background: Although maternal and newborn mortality have decreased 44 and 46% respectively between 1990 and 2015, achievement of ambitious Sustainable Development Goal targets requires accelerated progress. Mortality reduction requires a renewed focus on the continuum of maternal and newborn care from the household to the health facility. Although barriers to accessing skilled care are documented for specific contexts, there is a lack of systematic evidence on how women and families identify maternal and newborn illness and make decisions and subsequent care-seeking patterns. The focus of this multi-country study was to identify and describe illness recognition, decision-making, and care-seeking patterns across various contexts among women and newborns who survived and died to ultimately inform programmatic priorities moving forward. Methods: This study was conducted in seven countries\u2014Ethiopia, Tanzania, Uganda, Nigeria, India, Indonesia, and Nepal. Mixed-methods were utilized including event narratives (group interviews), in-depth interviews (IDIs), focus group discussions (FDGs), rapid facility assessments, and secondary analyses of existing program data. A common protocol and tools were developed in collaboration with study teams and adapted for each site, as needed. Sample size was a minimum of five cases of each type (e.g., perceived postpartum hemorrhage, maternal death, newborn illness, and newborn death) for each study site, with a total of 84 perceived PPH, 45 maternal deaths, 83 newborn illness, 55 newborn deaths, 64 IDIs/FGDs, and 99 health facility assessments across all sites. Analysis included coding within and across cases, identifying broad themes on recognition of illness, decision-making, and patterns of care seeking, and corresponding contextual factors. Technical support was provided throughout the process for capacity building, quality assurance, and consistency across sites. Conclusion: This study provides rigorous evidence on how women and families recognize and respond to maternal and newborn illness. By using a common methodology and tools, findings not only were site-specific but also allow for comparison across contexts

Immunological assays for chemokine detection in in-vitro culture of CNS cells

Herein we review the various methods currently in use for determining the expression of chemokines by CNS cells in vitro. Chemokine detection assays are used in conjuction with one another to provide a comprehensive, biologically relevant assessment of the chemokines which is necessary for correct data interpretation of a specific observed biological effect. The methods described include bioassays for soluble chemokine receptors, RNA extraction, RT-PCR, Real - time quantitative PCR, gene array analysis, northern blot analysis, Ribonuclease Protection assay, Flow cytometry, ELISPOT, western blot analysis, and ELISA. No single method of analysis meets the criteria for a comprehensive, biologically relevant assessment of the chemokines, therefore more than one assay might be necessary for correct data interpretation, a choice that is based on development of a scientific rationale for the method with emphasis on the reliability and relevance of the method

AN ANALYSIS OF THE PROXIMATE DETERMINANTS OF FERTILITY IN SUB-SAHARAN AFRICA WITH A FOCUS ON INDUCED ABORTION

Background: The fertility transition in sub-Saharan Africa (SSA) has been slower than in other regions of the world largely due to lack of rapid increase in contraceptive prevalence and the associated decline of unmet need for family planning. To study how fertility has changed across SSA, this dissertation will analyze the contributions of the proximate determinants of fertility to overall fertility decline by country, and undertake focal studies on one of the main proximate determinants – induced abortion. Methods: The data used for the analysis in this dissertation comes from the Demographic and Health Surveys. A total of 105 surveys covering 39 countries are used from SSA and seven non-SSA comparator countries. The first analytic paper calculates each of the proximate determinants using Bongaarts’ framework (1978, 1982) and decomposes change in fertility by country by the four major determinants. The second paper uses a country fixed effects multivariate linear regression model to determine the association between unmet need for family planning and induced abortion for all countries of SSA with available data, testing two different estimators of total abortion rate. The third analytic paper uses regression discontinuity design to isolate the impact of change in the abortion law in Ethiopia in 2005 on the incidence of reported pregnancy termination. Results: Most of the decline in fertility in SSA Is attributable to increases in the proportions of women unmarried, and to a lesser extent, increases in contraceptive use. A positive association exists between unmet need and total abortion rate for countries of SSA though the pathways inherent in this relationship are still unclear. And finally, the regression analysis in the third paper shows that the incidence of reported pregnancy termination neither increases nor decreases as a direct result of the change in abortion law in Ethiopia. Conclusion: Family planning programs should be further strengthened to reduce unmet need. Data on induced abortion need to be better collected and/or estimated. Loosening of abortion laws do not necessarily lead to greater uptake of abortion

Summary findings from a mixed methods study on identifying and responding to maternal and newborn illness in seven countries: implications for programs

Abstract Background There is a lack of systematic information documenting recognition of potentially life-threatening complications and decisions to seek care, as well as reaching care and the specific steps in that process. In response to this gap in knowledge, a multi-country mixed methods study was conducted to illuminate the dynamics driving Delays 1 and 2 across seven countries for maternal and newborn illness and death. Methods A common protocol and tools were developed, adapted by each of seven study teams depending on their local context (Ethiopia, India, Indonesia, Nigeria, Tanzania, Uganda, and Nepal). Maternal and newborn illness, and maternal and newborn death cases were included. Trained interviewers conducted event narratives to elicit and document a detailed sequence of actions, from onset of symptoms to the resolution of the problem. Event timelines were constructed, and in-depth interviews with key informants in the community were conducted. Transcripts were coded and analyzed for common themes corresponding to the three main domains of recognition, decision-making, and care-seeking. Results Maternal symptom recognition and decision-making to seek care is faster than for newborns. Perceived cause of the illness (supernatural vs. biological) influences the type of care sought (spiritual/traditional vs. formal sector, skilled). Mothers, fathers, and other relatives tend to be the decision-makers for newborns while husbands and elder females make decisions for maternal cases. Cultural norms such as confinement periods and perceptions of newborn vulnerability result in care being brought in to the home. Perceived and actual poor quality of care was repeatedly experienced by families seeking care. Conclusion The findings link to three action points: (1) messaging around newborn illness needs to reinforce a sense of urgency and the need for skilled care regardless of perceived cause; (2) targeted awareness building around specific maternal danger signs that are not currently recognized and where quality care is available is needed; and (3) designing appropriate contextualized messages. This research links to and supports a number of current global initiatives such as Ending Preventable Maternal Mortality, the Every Newborn Action Plan, the WHO Quality of Care framework, and the WHO guidelines on simplified management of newborn sepsis at the community level. This type of research is invaluable for designing programs to improve maternal and newborn survival to achieve ambitious global targets