Long-Term storage of Tularemia Agent strains

Keeping microorganisms alive, saving initial practically vital properties, is a major issue in microbiology. Objective of this research was to study preservation of the key biological properties of lyophilized cultures of 18 F. tularensis strains of different subspecies from the cultures collection of the «48 Central Research Institute» of the Ministry of Defense of the Russian Federation under varying conditions of preliminary preparation (including animalization through the guinea pigs’ body or without it). The microbe storage life at below-freezing temperatures ranged from 5 to 50 years. Materials and methods. The survival of microbes in the process of drying and subsequent storage was determined in accordance with the commonly-accepted methods; morphological, cultural, biochemical, antigenic, and pathogenic properties of F. tularensis strain cultures with different storage term and conditions for preparation to lyophilization were assessed and compared with datasheet specification. The results and conclusions. The research showed that under preliminary stabilization of F. tularensis strain properties using animalization, the storage in lyophilized state at below-freezing temperatures under vacuum for up to 50 years or less protected them from changes in phenotypic properties. At the same time, after lyophilization of cultures, which were repeatedly sub-cultured on dense nutrient media without initial signs of properties’ breach, phenomena of dissociation, the decrease in titers of cultures in the agglutination reaction from 2 to 4 times, and an increase in LD50 index by times were found

MANUFACTURING OF HYBRIDOMAS-PRODUCERS OF MONOCLONAL ANTIBODIES TO BRUCELLOSIS AGENT ANTIGENS

Objective of study is to prepare hybridomas-producers of monoclonal antibodies to brucellosis agent antigens. Materials and methods. B. abortus, B. melitensis, B. suis strains from the State collection of microorganisms of the 48th Central Research Institute Affiliated Branch and BALB/c mice. Hybridization was performed as described by G.Kohler and C.Milstein in modification by Fazekas De St. and Scheidegger D. The study of specific activity of immune sera, hybridoma supernatants, ascites fluid, and monoclonal antibody preparations was performed using ELISA. Results and conclusions. Obtained and characterized have been hybridomas-producers of monoclonal antibodies to specific antigens of brucellosis agent. They are active and stable antibody producers in the repeated passaging both, in vitro and in vivo. Obtained have also been the ascites fluid and preparations of monoclonal antibodies of brucellosis agent. Carried out has been substantiated selection of antibodies which could provide for the most sensitive ELISA. It is established that the monoclonal antibodies produced by hybridomas 232B6H7, 232G12F7, 233B2C5 in combination with brucellosis rabbit immunoglobulins allow for the identification of microbial cells of type strains of various Brucella species in concentrations ranging from 0,25·106 mc·sm–3 up to 1,0·106 mc·sm–3 and gave negative results with cultures of heterologous microorganisms in the contents of 1,0·108 mc·sm–3. Hybridomas-producers of monoclonal antibodies are planned to be used for the construction and manufacturing of immunodetection test-systems

Development of Immuno-Enzymatic Monoclonal Tests-Systems for the Detection of Glanders and Melioidosis Agents

Objective of the study was the development of immune-enzymatic monoclonal test-kit for detecting glanders and melioidosis agents. Materials and methods. We used microbial cultures and hybrid cell lines obtained from the collection of the «48th Central Research Institute» of the Ministry of Defense of the Russian Federation. Hybridoma cells were incubated in the peritoneal cavity of BALB/c mice. Preparations of glanders and melioidosis monoclonal antibodies were isolated from the ascetic fluids through precipitation with ammonium sulfate and purification by means of ion-exchange chromatography. Specific components of the test-kits were subjected to freeze drying in corresponding protective media. Study of diagnostic properties of the developed test systems was performed using ELISA. Results and conclusions. We have obtained preparations of monoclonal antibodies in vivo, as well as isolated and purified immunoglobulins from ascetic fluids. We also selected the pairs of monoclonal antibodies for manufacturing specific components. Experimental series of immune-enzymatic monoclonal test-systems allowing for specific detection of glanders and melioidosis causative agents in concentrations ranging from 0.5·106 CFU/ml and higher were made. The absence of cross-reactivity with closely related saprophytes and heterologous microorganisms in concentrations of 1,0·108 CFU/ml was shown. Demonstrated was the possibility in principle to differentiate between Burkholderia malleiand Burkholderia pseudomallei using ELISA. Test systems are promising for follow up state registration as medical products for in vitro diagnostics

Manufacturing of Hybridomas-Producers of Monoclonal Anti-Bodies to Tularemia Agent Antigens

Carried out have been two experimental studies on hybridization of mouse myeloma cells and lymphocytes of BALB/c mice immunized with inactivated microbe Francisella tularensis cultures. As a result obtained have been hybridomas-producers of monoclonal antibodies (MAb) specific to the antigens of tularemia agent. Evaluated have been the prospects of its application for the detection of the agent under discussion using enzyme-linked immunoassay. Established is the fact that monoclonal antibodies produced by 31G1F10, 32E5D3, 35B11C8, 36C2F11 hybridomas make it possible to identify microbe cells of various tularemia agent strains when concentrated up to 0.5·106 mc/sm3, and do not interact with cultures of heterologous microorganisms when concentrated to 1.0·108 mc/sm3, which testifies to their specificity. These MAb are planned to be used for the construction of immune-enzyme and immune-chromatographic test-systems designed for tularemia agent detection

Development of the Immuno-Enzyme Test-System for the Detection of <i>Legionella pheumophila</i>, Serogroup I

Developed is the highly sensitive and specific immuno-enzyme test-system, which is perspective for the detection of L. pneumophilia, serogroup 1. Isolated are the three hybrid cell lines that secrete monoclonal antibodies to specific epitopes of L. pneumophilia, serogroup 1 lipopolysaccharide antigen. Hyper immune rabbit sera, characterized by highly specific activity and specificity, are obtained using lipopolysaccharide antigen

Measuring universal health coverage based on an index of effective coverage of health services in 204 countries and territories, 1990–2019 : a systematic analysis for the Global Burden of Disease Study 2019

Background: Achieving universal health coverage (UHC) involves all people receiving the health services they need, of high quality, without experiencing financial hardship. Making progress towards UHC is a policy priority for both countries and global institutions, as highlighted by the agenda of the UN Sustainable Development Goals (SDGs) and WHO's Thirteenth General Programme of Work (GPW13). Measuring effective coverage at the health-system level is important for understanding whether health services are aligned with countries' health profiles and are of sufficient quality to produce health gains for populations of all ages. Methods: Based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we assessed UHC effective coverage for 204 countries and territories from 1990 to 2019. Drawing from a measurement framework developed through WHO's GPW13 consultation, we mapped 23 effective coverage indicators to a matrix representing health service types (eg, promotion, prevention, and treatment) and five population-age groups spanning from reproductive and newborn to older adults (>= 65 years). Effective coverage indicators were based on intervention coverage or outcome-based measures such as mortality-to-incidence ratios to approximate access to quality care; outcome-based measures were transformed to values on a scale of 0-100 based on the 2.5th and 97.5th percentile of location-year values. We constructed the UHC effective coverage index by weighting each effective coverage indicator relative to its associated potential health gains, as measured by disability-adjusted life-years for each location-year and population-age group. For three tests of validity (content, known-groups, and convergent), UHC effective coverage index performance was generally better than that of other UHC service coverage indices from WHO (ie, the current metric for SDG indicator 3.8.1 on UHC service coverage), the World Bank, and GBD 2017. We quantified frontiers of UHC effective coverage performance on the basis of pooled health spending per capita, representing UHC effective coverage index levels achieved in 2019 relative to country-level government health spending, prepaid private expenditures, and development assistance for health. To assess current trajectories towards the GPW13 UHC billion target-1 billion more people benefiting from UHC by 2023-we estimated additional population equivalents with UHC effective coverage from 2018 to 2023. Findings: Globally, performance on the UHC effective coverage index improved from 45.8 (95% uncertainty interval 44.2-47.5) in 1990 to 60.3 (58.7-61.9) in 2019, yet country-level UHC effective coverage in 2019 still spanned from 95 or higher in Japan and Iceland to lower than 25 in Somalia and the Central African Republic. Since 2010, sub-Saharan Africa showed accelerated gains on the UHC effective coverage index (at an average increase of 2.6% [1.9-3.3] per year up to 2019); by contrast, most other GBD super-regions had slowed rates of progress in 2010-2019 relative to 1990-2010. Many countries showed lagging performance on effective coverage indicators for non-communicable diseases relative to those for communicable diseases and maternal and child health, despite non-communicable diseases accounting for a greater proportion of potential health gains in 2019, suggesting that many health systems are not keeping pace with the rising non-communicable disease burden and associated population health needs. In 2019, the UHC effective coverage index was associated with pooled health spending per capita (r=0.79), although countries across the development spectrum had much lower UHC effective coverage than is potentially achievable relative to their health spending. Under maximum efficiency of translating health spending into UHC effective coverage performance, countries would need to reach $1398 pooled health spending per capita (US$ adjusted for purchasing power parity) in order to achieve 80 on the UHC effective coverage index. From 2018 to 2023, an estimated 388.9 million (358.6-421.3) more population equivalents would have UHC effective coverage, falling well short of the GPW13 target of 1 billion more people benefiting from UHC during this time. Current projections point to an estimated 3.1 billion (3.0-3.2) population equivalents still lacking UHC effective coverage in 2023, with nearly a third (968.1 million [903.5-1040.3]) residing in south Asia. Interpretation: The present study demonstrates the utility of measuring effective coverage and its role in supporting improved health outcomes for all people-the ultimate goal of UHC and its achievement. Global ambitions to accelerate progress on UHC service coverage are increasingly unlikely unless concerted action on non-communicable diseases occurs and countries can better translate health spending into improved performance. Focusing on effective coverage and accounting for the world's evolving health needs lays the groundwork for better understanding how close-or how far-all populations are in benefiting from UHC

Anemia prevalence in women of reproductive age in low- and middle-income countries between 2000 and 2018

Anemia is a globally widespread condition in women and is associated with reduced economic productivity and increased mortality worldwide. Here we map annual 2000–2018 geospatial estimates of anemia prevalence in women of reproductive age (15–49 years) across 82 low- and middle-income countries (LMICs), stratify anemia by severity and aggregate results to policy-relevant administrative and national levels. Additionally, we provide subnational disparity analyses to provide a comprehensive overview of anemia prevalence inequalities within these countries and predict progress toward the World Health Organization’s Global Nutrition Target (WHO GNT) to reduce anemia by half by 2030. Our results demonstrate widespread moderate improvements in overall anemia prevalence but identify only three LMICs with a high probability of achieving the WHO GNT by 2030 at a national scale, and no LMIC is expected to achieve the target in all their subnational administrative units. Our maps show where large within-country disparities occur, as well as areas likely to fall short of the WHO GNT, offering precision public health tools so that adequate resource allocation and subsequent interventions can be targeted to the most vulnerable populations

Measuring the availability of human resources for health and its relationship to universal health coverage for 204 countries and territories from 1990 to 2019: a systematic analysis for the Global Burden of Disease Study 2019

Background: Human resources for health (HRH) include a range of occupations that aim to promote or improve human health. The UN Sustainable Development Goals (SDGs) and the WHO Health Workforce 2030 strategy have drawn attention to the importance of HRH for achieving policy priorities such as universal health coverage (UHC). Although previous research has found substantial global disparities in HRH, the absence of comparable cross-national estimates of existing workforces has hindered efforts to quantify workforce requirements to meet health system goals. We aimed to use comparable and standardised data sources to estimate HRH densities globally, and to examine the relationship between a subset of HRH cadres and UHC effective coverage performance. Methods: Through the International Labour Organization and Global Health Data Exchange databases, we identified 1404 country-years of data from labour force surveys and 69 country-years of census data, with detailed microdata on health-related employment. From the WHO National Health Workforce Accounts, we identified 2950 country-years of data. We mapped data from all occupational coding systems to the International Standard Classification of Occupations 1988 (ISCO-88), allowing for standardised estimation of densities for 16 categories of health workers across the full time series. Using data from 1990 to 2019 for 196 of 204 countries and territories, covering seven Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) super-regions and 21 regions, we applied spatiotemporal Gaussian process regression (ST-GPR) to model HRH densities from 1990 to 2019 for all countries and territories. We used stochastic frontier meta-regression to model the relationship between the UHC effective coverage index and densities for the four categories of health workers enumerated in SDG indicator 3.c.1 pertaining to HRH: physicians, nurses and midwives, dentistry personnel, and pharmaceutical personnel. We identified minimum workforce density thresholds required to meet a specified target of 80 out of 100 on the UHC effective coverage index, and quantified national shortages with respect to those minimum thresholds. Findings: We estimated that, in 2019, the world had 104·0 million (95% uncertainty interval 83·5–128·0) health workers, including 12·8 million (9·7–16·6) physicians, 29·8 million (23·3–37·7) nurses and midwives, 4·6 million (3·6–6·0) dentistry personnel, and 5·2 million (4·0–6·7) pharmaceutical personnel. We calculated a global physician density of 16·7 (12·6–21·6) per 10 000 population, and a nurse and midwife density of 38·6 (30·1–48·8) per 10 000 population. We found the GBD super-regions of sub-Saharan Africa, south Asia, and north Africa and the Middle East had the lowest HRH densities. To reach 80 out of 100 on the UHC effective coverage index, we estimated that, per 10 000 population, at least 20·7 physicians, 70·6 nurses and midwives, 8·2 dentistry personnel, and 9·4 pharmaceutical personnel would be needed. In total, the 2019 national health workforces fell short of these minimum thresholds by 6·4 million physicians, 30·6 million nurses and midwives, 3·3 million dentistry personnel, and 2·9 million pharmaceutical personnel. Interpretation: Considerable expansion of the world's health workforce is needed to achieve high levels of UHC effective coverage. The largest shortages are in low-income settings, highlighting the need for increased financing and coordination to train, employ, and retain human resources in the health sector. Actual HRH shortages might be larger than estimated because minimum thresholds for each cadre of health workers are benchmarked on health systems that most efficiently translate human resources into UHC attainment