Novel human recombinant N-acetylgalactosamine-6-sulfate sulfatase produced in a glyco-engineered Escherichia coli strain.

Mucopolysaccharidosis IVA (MPS IVA) is a lysosomal storage disease caused by mutations in the gene encoding the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS), resulting in the accumulation of keratan sulfate (KS) and chondroitin-6-sulfate (C6S). Previously, it was reported the production of an active human recombinant GALNS (rGALNS) in E. coli BL21(DE3). However, this recombinant enzyme was not taken up by HEK293 cells or MPS IVA skin fibroblasts. Here, we leveraged a glyco-engineered E. coli strain to produce a recombinant human GALNS bearing the eukaryotic trimannosyl core N-glycan, Man3GlcNAc2 (rGALNSoptGly). The N-glycosylated GALNS was produced at 100 mL and 1.65 L scales, purified and characterized with respect to pH stability, enzyme kinetic parameters, cell uptake, and KS clearance. The results showed that the addition of trimannosyl core N-glycans enhanced both protein stability and substrate affinity. rGALNSoptGly was capture through a mannose receptor-mediated process. This enzyme was delivered to the lysosome, where it reduced KS storage in human MPS IVA fibroblasts. This study demonstrates the potential of a glyco-engineered E. coli for producing a fully functional GALNS enzyme. It may offer an economic approach for the biosynthesis of a therapeutic glycoprotein that could prove useful for MPS IVA treatment. This strategy could be extended to other lysosomal enzymes that rely on the presence of mannose N-glycans for cell uptake.

Are Contraceptive Method Preferences Stable? Measuring Change in the Preferred Method among Kenyan Women.

Contraceptive preferences are important for reproductive outcomes, such as contraceptive continuation and pregnancy. Current approaches to measuring reproductive preferences in population surveys are limited to exploring only fertility preferences and implicitly assume that contracepting people are using a method they want. We know that people change their fertility preferences over the life course as a response to life events, but there is no information about changes in contraceptive preferences, given the limited evidence about the measurement and distribution of contraceptive preferences. In this study, we examined the extent of change in women's contraceptive preferences over one year and identified characteristics associated with this change in Kenya using three rounds of nationally representative longitudinal data. Over one year, 18 percent of contraceptive users and 46 percent of contraceptive nonusers reported changes in their preferred contraceptive. Experiencing a pregnancy or birth and changes in marital status were associated with changes in contraceptive preferences for users and nonusers. We found that contraceptive preferences are dynamic, suggesting that family planning programs should ensure people's access to various methods to respond to women's changing circumstances and preferences.

Demographic dividend-favorable policy environment in two pre-dividend African nations: review of national policies and prospects for policy amendments in Nigeria and Tanzania.

BACKGROUND: In collaboration with local partners, we reviewed 18 national policy documents across two sub-Saharan African countries identified as pre-dividend nations by the World Bank in 2017: Nigeria and Tanzania. Our aim was to assess national policies in pre-dividend countries and to determine whether national strategies were primed to capitalize on changing demographic structures, maximally attain the demographic dividend, and augment socio-economic growth. METHODS: We conducted policy reviews by focusing on five key sectors of the Gates Institute Demographic Dividend Framework: Family Planning, Maternal and Child Health, Education, Women's Empowerment, and Labor Market. This framework was developed as a tool for countries to apply targeted policies for accelerating the demographic dividend based on their demographic structure. For each component we used a comprehensive list of indicators, defined via a systematic literature review, through which we assessed national policies aimed at maximizing the demographic dividend. RESULTS: Between the two countries, we observed persistent gaps in policies targeting family planning. Although more comprehensive, policies addressing maternal and child health, education, women's empowerment, and labor market still lagged in their specificity and measurability. We identified specific policy amendments and alternatives that Nigeria and Tanzania could consider to mitigate these gaps. We also stress the importance of designing measurable policy initiatives across sectors. CONCLUSIONS: Based on these recommendations, as Nigeria, Tanzania, and other pre-dividend nations start experiencing rapid demographic changes, they may consider implementing routine policy reviews to strengthen policies across the five key sectors and harness the benefits of a demographic dividend.

Development of the Demographic Dividend Effort Index, a novel tool to measure existing efforts to create a favourable environment to harness a demographic dividend: results from an experts' survey from six sub-Saharan African countries.

OBJECTIVE: This study aimed to develop a tool to measure the extent of national efforts in policies, services, research and programmes implemented to cultivate and harness the benefits of a potential demographic dividend in six sub-Saharan African countries. DESIGN: The survey was self-administered online using the SurveyMonkey platform. The survey questionnaire covered six key sectors: family planning, maternal and child health, education, women's empowerment, labour market, and governance and economic institution. Each sector-specific questionnaire was structured around five practice domains: policymaking, services and programmes, advocacy, research and civil society. Each item was scored from 1 to 10. Factor analysis was used to select the items to be retained for final score estimation. Simple averages were computed to estimate sectoral and domain scores and overall country scores were estimated using weighted country mean scores. Internal consistency, construct validity and reliability were examined using factor analysis and Cronbach's alpha. SETTING: Sub-Saharan Africa. PARTICIPANTS: A total of 440 knowledgeable informants from six countries; namely, Ethiopia (73), Kenya (69), Nigeria (67), Rwanda (54), Senegal (81) and Tanzania (96). RESULTS: Based on the results from factor analysis, 38 items were dropped from the analysis and Cronbach's alpha results ranged from 0.84 to 0.98 across domains. The overall demographic dividend effort index (DDEI) scores ranged between 5.4 (95% CI 5.1 to 5.8) in Ethiopia to 7.7 (95% CI 7.5 to 8.0) in Rwanda. In most countries, the disaggregated scores by sector revealed low scores in the labour market and women's empowerment. CONCLUSION: The DDEI scores highlight important gaps in key health and development sectors. The DDEI proved to be a reliable and internally consistent tool for effort measurement in key demographic dividend sectors. The DDEI can serve as a self-evaluation tool for local actors and may complement existing quantitative tools such as the Global Gender Gap and the Human Capital Index.

Everyday capabilities were a path to resilience during COVID-19: a case study of five countries.

COVID-19 demanded urgent responses by all countries, with wide variations in the scope and sustainability of those responses. Scholarship on resilience has increasingly emphasized relational considerations such as norms and power and how they influence health systems' responses to evolving challenges. In this study, we explored what influenced countries' national pandemic responses over time considering a country's capacity to test for COVID-19. To identify countries for inclusion, we used daily reports of COVID-19 cases and testing from 184 countries between 21st January 2020 and 31st December 2020. Countries reporting test data consistently and for at least 105 days were included, yielding a sample of 52 countries. We then sampled five countries representing different geographies, income levels and governance structures (Belgium, Ethiopia, India, Israel and Peru) and conducted semi-structured key informant interviews with stakeholders working in, or deeply familiar with, national responses. Across these five countries, we found that existing health systems capacities and political leadership determined how responses unfolded, while emergency plans or pandemic preparedness documents were not fit-for-purpose. While all five countries were successful at reducing COVID-19 infections at a specific moment in the pandemic, political economy factors complicated the ability to sustain responses, with all countries experiencing larger waves of the virus in 2021 or 2022. Our findings emphasize the continued importance of foundational public health and health systems capacities, bolstered by clear leadership and multisectoral coordination functions. Even in settings with high-level political leadership and a strong multisectoral response, informants wished that they-and their country's health system-were more prepared to address the pandemic and maintain an effective response over time. Our findings challenge emergency preparedness as the dominant frame in pandemic preparedness and call for a continued emphasis on health systems strengthening to respond to future health shocks and a pandemic moving to endemic status.

The relationship between client dissatisfaction and contraceptive discontinuation among urban family planning clients in three sub-Saharan African countries.

Although researchers and practitioners have suggested that the quality of family planning services impacts contraceptive discontinuation, establishing a causal relationship has been challenging, primarily due to data limitations and a lack of agreement on how to measure quality. This longitudinal study estimated the relationship of the dissatisfaction with family planning services on contraceptive discontinuation for a sample of 797 female clients who sought family planning services at urban facilities across Kenya, Nigeria, and Burkina Faso. Clients who sought family planning services were first interviewed in person at private and public health facilities and received a follow-up phone interview four to six months later. In our sample, 18.2% of clients who were using a modern contraceptive at baseline stopped using it by follow-up. At baseline, nearly 14% of clients reported experiencing a problem with service convenience, nearly 12% with the availability of medicines and contraceptives, and nearly 6% with facility cleanliness and/or staff treatment. We hypothesized that client dissatisfaction with the family planning services received informed their decision to discontinue contraception and estimated univariate and bivariate probit regression models, controlling for individual and health facility characteristics. We found that client's perceptions of staff treatment and facility cleanliness informed their expectations about service and contraceptive standards, affecting subsequent contraceptive discontinuation. The difference in the probability of discontinuing contraception was 8.2 percentage-points between dissatisfied and satisfied clients. Examining client dissatisfaction with family planning services can inform the family planning community on needed improvements to increase contraceptive adherence for women in need, which can prevent unplanned pregnancies and unwanted births in the long run.

Mucocutaneous manifestations and their relationship with CD4 T-lymphocyte count in hospitalized patients infected with the human immunodeficiency virus (HIV) in Medellín, Colombia / Manifestaciones mucocutáneas y su relación con el recuento de linfocitos T CD4 en pacientes infectados con el virus de inmunodeficiencia humana hospitalizados en Medellín, Colombia

Introduction: About 80-95% of patients infected with the human immunodeficiency virus (HIV) develop skin manifestations, which are markers of the patients' immune status. Objective: To describe the dermatologic manifestations and the clinical and sociodemographic factors of hospitalized patients diagnosed with HIV and their correlation with CD4 T-lymphocyte count. Materials and methods: We conducted an observational, cross-sectional, and retrospective study of the medical records of 227 adult patients with HIV diagnosis evaluated by dermatology in a hospital in Medellín, Colombia. Results: We included 227 patient records with 433 dermatologic manifestations, 64.4% of them infectious. The most frequent manifestations were oral candidiasis, condylomata acuminata, and drug reactions. Moreover, a statistically significant relationship was found between disseminated herpes zoster virus and secondary syphilis with a CD4 count between 200-499 cells/mm3 (p=0.04 and 0.028, respectively). There was also a statistically significant relationship between oral candidiasis and a CD4 count of less than 100 cells/mm3 (p=0.008). Conclusions: The relationship between disseminated herpes zoster with CD4 between 200-499 cells/mm3 suggests that, despite having high CD4 counts, severe forms of the disease may occur due to possible T-cell dysfunction and depletion of the immune system. Additionally, the relationship between oral candidiasis and CD4 less than 100 cells/mm3 indicates the potential role of oral candidiasis as an essential marker of weakened immune status in HIV patients.

Introducción. Entre el 80 y el 95 % de los pacientes infectados por el virus de inmunodeficiencia humana (HIV) desarrollan manifestaciones en la piel que sirven como marcadores de su estado inmunológico. Objetivos. Describir las manifestaciones dermatológicas y los factores clínicos y sociodemográficos de los pacientes hospitalizados con diagnóstico de HIV y su correlación con el recuento de linfocitos T CD4. Materiales y métodos. Se hizo un estudio observacional de corte transversal y retrospectivo a partir del registro de las historias clínicas de 227 pacientes mayores de edad con diagnóstico de HIV, evaluados por dermatología en un hospital de Medellín, Colombia. Resultados. Los 227 registros daban cuenta de 433 manifestaciones dermatológicas, el 64,4 % de ellas infecciosas. Las tres manifestaciones más frecuentes fueron candidiasis oral, condilomas acuminados y reacciones a medicamentos. Se encontró una relación estadísticamente significativa entre el virus del herpes zóster (HZ) diseminado y la sífilis secundaria, con un recuento de CD4 entre 200 y 499 células/mm3 (p=0,04 y 0,028, respectivamente), y entre la candidiasis oral y un recuento de CD4 menor de 100 células/mm3 (p=0,008). Conclusiones. La relación entre el herpes zóster diseminado y un recuento de CD4 entre 200 y 499 células/mm3 sugiere que, a pesar de los recuentos altos, se pueden presentar formas graves de la enfermedad debido a una posible disfunción de las células T y el agotamiento del sistema inmunológico. La relación entre la candidiasis oral y un recuento de CD4 menor de 100 células/mm3 plantea la posibilidad de considerar esta infección micótica como un marcador importante de debilitamiento inmunológico de los pacientes con HIV.

Past local government health spending was not correlated with COVID-19 control in US counties.

CONTEXT: Wide variation in state and county health spending prior to 2020 enables tests of whether historically better state and locally funded counties achieved faster control over COVID-19 in the first 6 months of the pandemic in the Unites States prior to federal supplemental funding. OBJECTIVE: We used time-to-event and generalized linear models to examine the association between pre-pandemic state-level public health spending, county-level non-hospital health spending, and effective COVID-19 control at the county level. We include 2,775 counties that reported 10 or more COVID-19 cases between January 22, 2020, and July 19, 2020, in the analysis. MAIN OUTCOME MEASURE: Control of COVID-19 was defined by: (i) elapsed time in days between the 10th case and the day of peak incidence of a county's local epidemic, among counties that bent their case curves, and (ii) doubling time of case counts within the first 30 days of a county's local epidemic for all counties that reported 10 or more cases. RESULTS: Only 26% of eligible counties had bent their case curve in the first 6 months of the pandemic. Government health spending at the county level was not associated with better COVID-19 control in terms of either a shorter time to peak in survival analyses, or doubling time in generalized linear models. State-level public spending on hazard preparation and response was associated with a shorter time to peak among counties that were able to bend their case incidence curves. CONCLUSIONS: Increasing resource availability for public health in local jurisdictions without thoughtful attention to bolstering the foundational capabilities inside health departments is unlikely to be sufficient to prepare the country for future outbreaks or other public health emergencies.

The wealth gradient and the effect of COVID-19 restrictions on income loss, food insecurity and health care access in four sub-Saharan African geographies.

INTRODUCTION: While there has been considerable analysis of the health and economic effects of COVID-19 in the Global North, representative data on the distribution and depth of social and economic impacts in Africa has been more limited. METHODS: We analyze household data collected prior to the COVID-19 pandemic and during the first wave of COVID in four African countries. We evaluate the short-term changes to household economic status and assess women's access to health care during the first wave of COVID-19 in nationally representative samples of women aged 15-49 in Kenya and Burkina Faso, and in sub-nationally representative samples of women aged 15-49 in Kinshasa, Democratic Republic of Congo and Lagos, Nigeria. We examine prevalence and distribution of household income loss, food insecurity, and access to health care during the COVID-19 lockdowns across residence and pre-pandemic wealth categories. We then regress pre-pandemic individual and household sociodemographic characteristics on the three outcomes. RESULTS: In three out of four samples, over 90% of women reported partial or complete loss of household income since the beginning of the coronavirus restrictions. Prevalence of food insecurity ranged from 17.0% (95% CI 13.6-20.9) to 39.8% (95% CI 36.0-43.7), and the majority of women in food insecure households reported increases in food insecurity during the COVID-19 restriction period. In contrast, we did not find significant barriers to accessing health care during COVID restrictions. Between 78·3% and 94·0% of women who needed health care were able successfully access it. When we examined pre-pandemic sociodemographic correlates of the outcomes, we found that the income shock of COVID-19 was substantial and distributed similarly across wealth groups, but food insecurity was concentrated among poorer households. Contrary to a-priori expectations, we find little evidence of women experiencing barriers to health care, but there is significant need for food support.

County health outcomes linkage to county spending on social services, building infrastructure, and law and order.

Will counties that reallocate money from law enforcement to social services improve subsequent markers of population wellbeing? In this study, we measure the association between county government spending across multiple sectors and Life Expectancy at Birth (LEB) in the U.S. using data from the U.S. Census Bureau. We constructed a Structural Equation Model to determine whether social expenditure, building infrastructure, and spending on law and order were positively or negatively associated with LEB three-years after initial spending. The analysis compared data between 2002-05 and 2007-10 and was stratified for urban and rural counties. In rural counties, a one-standard-deviation increase in social spending increased subsequent LEB by 0.58 (SE 0.16) and 0.36 (SE 0.16) years in 2005 and 2010, respectively. In urban counties, a one-standard-deviation increase in building infrastructure spending increased subsequent LEB by 1.14 (SE 0.51) and 1.05 (SE 0.49) years in 2005 and 2010, respectively. In 2002, a one-standard-deviation increase in law and order spending significantly decreased subsequent life expectancy, 2.2 (SE 1.27) and 0.46 (SE 0.13) years in urban and rural counties, respectively. Similarly, investments in building infrastructure for urban counties and social services for rural counties were associated with subsequently higher life expectancy three years later after initial investments.

Is Client Reporting on Contraceptive Use Always Accurate? Measuring Consistency and Change with a Multicountry Study.

The consistency of self-reported contraceptive use over short periods of time is important for understanding measurement reliability. We assess the consistency of and change in contraceptive use using longitudinal data from 9,390 urban female clients interviewed in DR Congo, India, Kenya, Niger, Nigeria, and Burkina Faso. Clients were interviewed in-person at a health facility and four to six months later by phone. We compared reports of contraceptive use at baseline with recall of baseline contraceptive use at follow-up. Agreement between these measures ranged from 59.1 percent in DR Congo to 84.4 percent in India. Change in both contraceptive method type (sterilization, long-acting, short-acting, nonuse) and use status (user, nonuser, discontinuer, adopter, switcher) was assessed comparing baseline to follow-up reports and retrospective versus current reports within the follow-up survey. More change in use was observed with panel reporting than within the cross section. The percent agreement between the two scenarios of change ranged from 64.8 percent in DR Congo to 84.5 percent in India, with cross-site variation. Consistently reported change in use status was highest for nonusers, followed by users, discontinuers, adopters, and switchers. Inconsistency in self-reported contraceptive use, even over four to six months, was nontrivial, indicating that studying measurement reliability of contraceptive use remains important.

Service delivery point and individual characteristics associated with the adoption of modern contraceptive: A multi-country longitudinal analysis.

BACKGROUND: Women who start using contraception ("adopters") are a key population for family planning goals, but little is known about characteristics that predict the adoption of contraception as opposed to current use. We used prospective data from women and facilities for five countries, (Democratic Republic of Congo, India, Kenya, Nigeria, and Burkina Faso) and identified baseline characteristics that predicted adoption of modern contraception in the short term. METHODS: We used data from the Performance Monitoring for Action (PMA) Agile Project. PMA Agile administered service delivery point (SDP) client exit interview (CEI) surveys in urban sites of these five countries. Female clients responding to the CEI were asked for phone numbers that were used for a phone follow-up survey approximately four months later. For our analysis, we used data from the SDP and CEI baseline surveys, and the phone follow up to compare women who start using contraception during this period with those who remain non-users. We used characteristics of the facility and the woman at baseline to predict her contraception adoption in the future. RESULTS: Discussing FP with a partner at baseline was associated with greater odds of adoption in DRC (OR 2.34; 95% CI 0.97-5.66), India (OR 2.27; 95% CI 1.05-4.93), and Kenya (OR 1.65; 95% CI 1.16-2.35). Women who discussed family planning with any staff member at the health facility had 1.72 greater odds (95% CI 1.13-2.67) of becoming an adopter in Nigeria. The odds of adoption were lower in Nigerian facilities that had a stockout (OR 0.66 95% CI 0.44-1.00) at baseline. Other characteristics associated with contraception adoption across settings were education, age, wealth, parity, and marital status. CONCLUSIONS: Characteristics of both the woman and the health facility were associated with adoption of modern contraception in the future. Some characteristics, like discussing family planning with a spouse, education, and parity, were associated with contraceptive adoption across settings. Other characteristics that predict contraceptive use, such as health facility measures, varied across countries.

The Role of Public Health Expenditures in COVID-19 control: Evidence from Local Governments in England.

For over 150 years the local health departments of England have been critical in controlling 19th and 20th century infectious epidemics. However, recent administrative changes have hollowed out their flexibility to serve communities. We use administrative data on past budgetary allocations per capita to public health departments at upper tier local areas (UTLAs) of England to examine whether public health funding levels were correlated with more rapid control of the first wave of the COVID-19 pandemic between March and July of 2020. The dependent variable was the number of days between a UTLA's 10th case of COVID-19 and the day when new cases per 100,000 peaked and began to decline. Our models controlled for regional socio-economic factors. We found no correlation between local public health expenditure and the speed of control of COVID-19. However, overall public expenditure allocated to improve local areas helped reduce time to reach peak. Contrary to expectation, more dense areas such as London experienced shorter duration. Higher income areas had more rapid success in accelerating the time of the first peak in the first wave of their local COVID-19 incidence. We contribute to understanding the impact of how public expenditure and socio-economic factors affect an epidemic.

Glutaminase isoforms expression switches microRNA levels and oxidative status in glioblastoma cells.

BACKGROUND: Glutaminase isoenzymes GLS and GLS2 play apparently opposing roles in cancer: GLS acts as an oncoprotein, while GLS2 (GAB isoform) has context specific tumour suppressive activity. Some microRNAs (miRNAs) have been implicated in progression of tumours, including gliomas. The aim was to investigate the effect of GLS and GAB expression on both miRNAs and oxidative status in glioblastoma cells. METHODS: Microarray profiling of miRNA was performed in GLS-silenced LN229 and GAB-transfected T98G human glioblastoma cells and their wild-type counterparts. Results were validated by real-time quantitative RT-PCR. Oxidative status and antioxidant enzymes were determined by spectrophotometric or fluorescence assays in GLS-silenced LN229 and T98G, and GAB-transfected LN229 and T98G. RESULTS: MiRNA-146a-5p, miRNA-140-3p, miRNA-21-5p, miRNA-1260a, and miRNA-92a-3p were downregulated, and miRNA-1246 was upregulated when GLS was knocked down. MiRNA-140-3p, miRNA-1246, miRNA-1260a, miRNA-21-5p, and miRNA-146a-5p were upregulated when GAB was overexpressed. Oxidative status (lipid peroxidation, protein carbonylation, total antioxidant capacity, and glutathione levels), as well as antioxidant enzymes (catalase, superoxide dismutase, and glutathione reductase) of silenced GLS glioblastoma cells and overexpressed GAB glioblastoma cells significantly changed versus their respective control glioblastoma cells. MiRNA-1246, miRNA-1260a, miRNA-146a-5p, and miRNA-21-5p have been characterized as strong biomarkers of glioblastoma proliferation linked to both GLS silencing and GAB overexpression. Total glutathione is a reliable biomarker of glioblastoma oxidative status steadily associated to both GLS silencing and GAB overexpression. CONCLUSIONS: Glutaminase isoenzymes are related to the expression of some miRNAs and may contribute to either tumour progression or suppression through certain miRNA-mediated pathways, proving to be a key tool to switch cancer proliferation and redox status leading to a less malignant phenotype. Accordingly, GLS and GAB expression are especially involved in glutathione-dependent antioxidant defence.

Performance monitoring and accountability: The Agile Project's protocol, record and experience.

The Performance Monitoring and Accountability 2020 (PMA2020) project implemented a multi-country sub-project called PMA Agile, a system of continuous data collection for a probability sample of urban public and private health facilities and their clients that began November 2017 and concluded December 2019.  The objective was to monitor the supply, quality and consumption of family planning services.  In total, across 14 urban settings, nearly 2300 health facilities were surveyed three to six times in two years and a total sample of 48,610 female and male clients of childbearing age were interviewed in Burkina Faso, Democratic Republic of Congo, India, Kenya, Niger and Nigeria.  Consenting female clients with access to a cellphone were re-interviewed by telephone after four months; two rounds of the client exit, and follow-up interviews were conducted in nearly all settings.  This paper reports on the PMA Agile data system protocols, coverage and early experiences.  An online dashboard is publicly accessible, analyses of measured trends are underway, and the data are publicly available.

Nuclear Translocation of Glutaminase GLS2 in Human Cancer Cells Associates with Proliferation Arrest and Differentiation.

Glutaminase (GA) catalyzes the first step in mitochondrial glutaminolysis playing a key role in cancer metabolic reprogramming. Humans express two types of GA isoforms: GLS and GLS2. GLS isozymes have been consistently related to cell proliferation, but the role of GLS2 in cancer remains poorly understood. GLS2 is repressed in many tumor cells and a better understanding of its function in tumorigenesis may further the development of new therapeutic approaches. We analyzed GLS2 expression in HCC, GBM and neuroblastoma cells, as well as in monkey COS-7 cells. We studied GLS2 expression after induction of differentiation with phorbol ester (PMA) and transduction with the full-length cDNA of GLS2. In parallel, we investigated cell cycle progression and levels of p53, p21 and c-Myc proteins. Using the baculovirus system, human GLS2 protein was overexpressed, purified and analyzed for posttranslational modifications employing a proteomics LC-MS/MS platform. We have demonstrated a dual targeting of GLS2 in human cancer cells. Immunocytochemistry and subcellular fractionation gave consistent results demonstrating nuclear and mitochondrial locations, with the latter being predominant. Nuclear targeting was confirmed in cancer cells overexpressing c-Myc- and GFP-tagged GLS2 proteins. We assessed the subnuclear location finding a widespread distribution of GLS2 in the nucleoplasm without clear overlapping with specific nuclear substructures. GLS2 expression and nuclear accrual notably increased by treatment of SH-SY5Y cells with PMA and it correlated with cell cycle arrest at G2/M, upregulation of tumor suppressor p53 and p21 protein. A similar response was obtained by overexpression of GLS2 in T98G glioma cells, including downregulation of oncogene c-Myc. Furthermore, human GLS2 was identified as being hypusinated by MS analysis, a posttranslational modification which may be relevant for its nuclear targeting and/or function. Our studies provide evidence for a tumor suppressor role of GLS2 in certain types of cancer. The data imply that GLS2 can be regarded as a highly mobile and multilocalizing protein translocated to both mitochondria and nuclei. Upregulation of GLS2 in cancer cells induced an antiproliferative response with cell cycle arrest at the G2/M phase.

Generating and capitalizing on the demographic dividend potential in sub-Saharan Africa: a conceptual framework from a systematic literature review.

Background: Africa will double its population by 2050 and more than half will be below age 25. The continent has a unique opportunity to boost its socioeconomic welfare. This systematic literature review aims to develop a conceptual framework that identifies policies and programs that have provided a favorable environment for generating and harnessing a demographic dividend. This framework can facilitate sub-Saharan African countries' understanding of needed actions to accelerate their demographic transition and capitalize on their demographic dividend potential. Methods: The search strategy was structured around three concepts: economic development, fertility, and sub-Saharan Africa. Databases used included PubMed and EconLit. An inductive approach was employed to expand the reference base further. Data were extracted using literature records following a checklist of items to include when reporting a systematic review suggested in the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) Statement. Results: The final review consisted of 78 peer-reviewed articles, ten reports from the gray literature, and one book. Data were categorized according to relevant demographic dividend typology: pre-dividend and early-dividend. The results from the literature review were synthesized into a framework consisting of five sectors for pre-dividend countries, namely 1) Governance and Economic Institutions, 2) Family Planning, 3) Maternal and Child Health, 4) Education, and 5) Women's Empowerment. An additional sector, 6) Labor Market, is added for early-dividend countries. These sectors must work together to attain a demographic dividend. Conclusions: A country's demographic transition stage must guide policy and programs. Most sub-Saharan African countries have prioritized job creation and employment for youth, yet their efforts to secure a productive labor market require preliminary and complementary investments in governance, family planning, maternal and child health, education, and women's empowerment. Creating a favorable policy environment for generating and capitalizing on a demographic dividend can support their stated goals for development.

Ulcers in leprosy patients, an unrecognized clinical manifestation: a report of 8 cases.

BACKGROUND: Leprosy is a chronic granulomatous infection caused by Mycobacterium leprae. It is a polymorphic disease with a wide range of cutaneous and neural manifestations. Ulcer is not a common feature in leprosy patients, except during reactional states, Lucio's phenomenon (LP), or secondary to neuropathies. CASES PRESENTATION: We report eight patients with multibacillary leprosy who presented specific skin ulcers as part of their main leprosy manifestation. Ulcers were mostly present on lower limbs (eight patients), followed by the upper limbs (three patients), and the abdomen (one patient). Mean time from onset of skin ulcers to diagnosis of leprosy was 17.4 months: all patients were either misdiagnosed or had delayed diagnosis, with seven of them presenting grade 2 disability by the time of the diagnosis. Reactional states, LP or neuropathy as potential causes of ulcers were ruled out. Biopsy of the ulcer was available in seven patients: histopathology showed mild to moderate lympho-histiocytic infiltrate with vacuolized histiocytes and intact isolated and grouped acid-fast bacilli. Eosinophils, vasculitis, vasculopathy or signs of chronic venous insufficiency were not observed. Skin lesions improved rapidly after multidrug therapy, without any concomitant specific treatment for ulcers. CONCLUSIONS: This series of cases highlights the importance of recognizing ulcers as a specific cutaneous manifestation of leprosy, allowing diagnosis and treatment of the disease, and therefore avoiding development of disabilities and persistence of the transmission chain of M. leprae.