Optimized Case Finding of tuberculosis among key populations in Ukraine. A follow up study.

INTRODUCTION: In 2021, there were 4 million tuberculosis (TB) cases that were not detected by health systems, globally. Many of those cases are among hard-to-reach populations or key population groups. An Optimized Case Finding (OCF) strategy was introduced in Ukraine to enhance case detection and identify those "missing" cases. OCF included screening of up to eight referred household and social network contacts of an index TB case. Following the OCF project implementation, TB detection and characteristics of index cases and contacts were assessed. METHODOLOGY: A cohort study using project data (July 2018 - April 2022) was conducted. RESULTS: In total 7,976 close contacts were engaged in the project from 1,028 index TB cases. Among the contacts, 507 were diagnosed with TB. The TB case detection was 6,356/100,000 and the number needed to investigate was 16. Multiple factors were identified as associated with TB detection including smoking, HIV, poverty, etc. About 90% of cases were identified at the initial screening of the contacts. OCF was proven to be 5.8 times more effective than the standard active case finding using household surveys and 106 times more effective than passive case finding in the general public. CONCLUSIONS: Our study demonstrated the effectiveness of OCF in detecting cases among key population groups and their social networks. We encourage adaptation and use of OCF by civil society organizations that already work with key vulnerable populations around the globe.

ER and SOCE Ca2+ signals are not required for directed cell migration in human microglia.

The central nervous system (CNS) is constantly surveilled by microglia, highly motile and dynamic cells deputed to act as the first line of immune defense in the brain and spinal cord. Alterations in the homeostasis of the CNS are detected by microglia that respond by migrating toward the affected area. Understanding the mechanisms controlling directed cell migration of microglia is crucial to dissect their responses to neuroinflammation and injury. We used a combination of pharmacological and genetic approaches to explore the involvement of calcium (Ca2+) signaling in the directed migration of induced pluripotent stem cell (iPSC)-derived microglia challenged with a purinergic stimulus. This approach mimics cues originating from injury of the CNS. Unexpectedly, simultaneous imaging of microglia migration and intracellular Ca2+ changes revealed that this phenomenon does not require Ca2+ signals generated from the endoplasmic reticulum (ER) and store-operated Ca2+ entry (SOCE) pathways. Instead, we find evidence that human microglial chemotaxis to purinergic signals is mediated by cyclic AMP in a Ca2+-independent manner. These results challenge prevailing notions, with important implications in neurological conditions characterized by perturbation in Ca2+ homeostasis.

Alteration of microglial metabolism and inflammatory profile contributes to neurotoxicity in a hiPSC-derived microglia model of frontotemporal dementia 3.

Frontotemporal dementia (FTD) is a common cause of early-onset dementia, with no current treatment options. FTD linked to chromosome 3 (FTD3) is a rare sub-form of the disease, caused by a point mutation in the Charged Multivesicular Body Protein 2B (CHMP2B). This mutation causes neuronal phenotypes, such as mitochondrial deficiencies, accompanied by metabolic changes and interrupted endosomal-lysosomal fusion. However, the contribution of glial cells to FTD3 pathogenesis has, until recently, been largely unexplored. Glial cells play an important role in most neurodegenerative disorders as drivers and facilitators of neuroinflammation. Microglia are at the center of current investigations as potential pro-inflammatory drivers. While gliosis has been observed in FTD3 patient brains, it has not yet been systematically analyzed. In the light of this, we investigated the role of microglia in FTD3 by implementing human induced pluripotent stem cells (hiPSC) with either a heterozygous or homozygous CHMP2B mutation, introduced into a healthy control hiPSC line via CRISPR-Cas9 precision gene editing. These hiPSC were differentiated into microglia to evaluate the pro-inflammatory profile and metabolic state. Moreover, hiPSC-derived neurons were cultured with conditioned microglia media to investigate disease specific interactions between the two cell populations. Interestingly, we identified two divergent inflammatory microglial phenotypes resulting from the underlying mutations: a severe pro-inflammatory profile in CHMP2B homozygous FTD3 microglia, and an "unresponsive" CHMP2B heterozygous FTD3 microglial state. These findings correlate with our observations of increased phagocytic activity in CHMP2B homozygous, and impaired protein degradation in CHMP2B heterozygous FTD3 microglia. Metabolic mapping confirmed these differences, revealing a metabolic reprogramming of the CHMP2B FTD3 microglia, displayed as a compensatory up-regulation of glutamine metabolism in the CHMP2B homozygous FTD3 microglia. Intriguingly, conditioned CHMP2B homozygous FTD3 microglia media caused neurotoxic effects, which was not evident for the heterozygous microglia. Strikingly, IFN-γ treatment initiated an immune boost of the CHMP2B heterozygous FTD3 microglia, and conditioned microglia media exposure promoted neural outgrowth. Our findings indicate that the microglial profile, activity, and behavior is highly dependent on the status of the CHMP2B mutation. Our results suggest that the heterozygous state of the mutation in FTD3 patients could potentially be exploited in form of immune-boosting intervention strategies to counteract neurodegeneration.

Astrocytic response mediated by the CLU risk allele inhibits OPC proliferation and myelination in a human iPSC model.

The C allele of rs11136000 variant in the clusterin (CLU) gene represents the third strongest known genetic risk factor for late-onset Alzheimer's disease. However, whether this single-nucleotide polymorphism (SNP) is functional and what the underlying mechanisms are remain unclear. In this study, the CLU rs11136000 SNP is identified as a functional variant by a small-scale CRISPR-Cas9 screen. Astrocytes derived from isogenic induced pluripotent stem cells (iPSCs) carrying the "C" or "T" allele of the CLU rs11136000 SNP exhibit different CLU expression levels. TAR DNA-binding protein-43 (TDP-43) preferentially binds to the "C" allele to promote CLU expression and exacerbate inflammation. The interferon response and CXCL10 expression are elevated in cytokine-treated C/C astrocytes, leading to inhibition of oligodendrocyte progenitor cell (OPC) proliferation and myelination. Accordingly, elevated CLU and CXCL10 but reduced myelin basic protein (MBP) expression are detected in human brains of C/C carriers. Our study uncovers a mechanism underlying reduced white matter integrity observed in the CLU rs11136000 risk "C" allele carriers.

Strengthening the Operational Research Capacity of National Tuberculosis Control Programs: Necessity or Luxury?

'How to get research into practice: first get practice into research [...].

CRISPR generation of CSF1R-G795A human microglia for robust microglia replacement in a chimeric mouse model.

Chimeric mouse models have recently been developed to study human microglia in vivo. However, widespread engraftment of donor microglia within the adult brain has been challenging. Here, we present a protocol to introduce the G795A point mutation using CRISPR-Cas9 into the CSF1R locus of human pluripotent stem cells. We also describe an optimized microglial differentiation technique for transplantation into newborn or adult recipients. We then detail pharmacological paradigms to achieve widespread and near-complete engraftment of human microglia. For complete details on the use and execution of this protocol, please refer to Chadarevian et al. (2023).1.

Novel Vaccine against Pathological Pyroglutamate-Modified Amyloid Beta for Prevention of Alzheimer's Disease.

Post-translationally modified N-terminally truncated amyloid beta peptide with a cyclized form of glutamate at position 3 (pE3Aß) is a highly pathogenic molecule with increased neurotoxicity and propensity for aggregation. In the brains of Alzheimer's Disease (AD) cases, pE3Aß represents a major constituent of the amyloid plaque. The data show that pE3Aß formation is increased at early pre-symptomatic disease stages, while tau phosphorylation and aggregation mostly occur at later stages of the disease. This suggests that pE3Aß accumulation may be an early event in the disease pathogenesis and can be prophylactically targeted to prevent the onset of AD. The vaccine (AV-1986R/A) was generated by chemically conjugating the pE3Aß3-11 fragment to our universal immunogenic vaccine platform MultiTEP, then formulated in AdvaxCpG adjuvant. AV-1986R/A showed high immunogenicity and selectivity, with endpoint titers in the range of 105-106 against pE3Aß and 103-104 against the full-sized peptide in the 5XFAD AD mouse model. The vaccination showed efficient clearance of the pathology, including non-pyroglutamate-modified plaques, from the mice brains. AV-1986R/A is a novel promising candidate for the immunoprevention of AD. It is the first late preclinical candidate which selectively targets a pathology-specific form of amyloid with minimal immunoreactivity against the full-size peptide. Successful translation into clinic may offer a new avenue for the prevention of AD via vaccination of cognitively unimpaired individuals at risk of disease.

Epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples, Ecuador, 2018.

Objective: To compare the epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples in Ecuador. Methods: A secondary analysis was done of data on bacteria isolated from inpatient and outpatient samples. Data were taken from the 2018 national antimicrobial resistance surveillance database of the National Reference Center for Antimicrobial Resistance. The variables included were: age, sex, inpatient versus outpatient setting, type of specimen, bacterial species identified, pattern of resistance to antibiotics, and geographic area. Results: Data from 57 305 bacterial isolates were included in the study: 48.8% were from hospitalized patients, 55.7% were from women, and 60.1% were from patients older than 45 years. Urine (42.9%) and blood (12.4%) were the most common clinical samples. Overall, 77.1% of bacterial isolates were gram-negative (83% and 71% in outpatients and inpatients, respectively). The most common gram-positive and gram-negative species were Staphylococcus aureus and Escherichia coli, respectively. Antimicrobial resistance levels were high (up to 80% for some antimicrobial drugs), and were higher in hospitalized patients compared with outpatients. A variety of carbapenemases were found to confer resistance to carbapenems (antibiotics of last resort) in gram-negative bacteria. Conclusions: The study findings provide an important baseline on antimicrobial resistance in Ecuador. This will allow the strengthening of guidelines of the surveillance system, the creation of public policies for standardization of laboratory methodologies, the proper handling of information, and the development of empirical therapy guidelines based on local epidemiology.

Epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples, Ecuador, 2018

[ABSTRACT]. Objective. To compare the epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples in Ecuador. Methods. A secondary analysis was done of data on bacteria isolated from inpatient and outpatient samples. Data were taken from the 2018 national antimicrobial resistance surveillance database of the National Refer- ence Center for Antimicrobial Resistance. The variables included were: age, sex, inpatient versus outpatient setting, type of specimen, bacterial species identified, pattern of resistance to antibiotics, and geographic area. Results. Data from 57 305 bacterial isolates were included in the study: 48.8% were from hospitalized patients, 55.7% were from women, and 60.1% were from patients older than 45 years. Urine (42.9%) and blood (12.4%) were the most common clinical samples. Overall, 77.1% of bacterial isolates were gram-negative (83% and 71% in outpatients and inpatients, respectively). The most common gram-positive and gram-negative species were Staphylococcus aureus and Escherichia coli, respectively. Antimicrobial resistance levels were high (up to 80% for some antimicrobial drugs), and were higher in hospitalized patients compared with outpatients. A variety of carbapenemases were found to confer resistance to carbapenems (antibiotics of last resort) in gram-negative bacteria. Conclusions. The study findings provide an important baseline on antimicrobial resistance in Ecuador. This will allow the strengthening of guidelines of the surveillance system, the creation of public policies for stan- dardization of laboratory methodologies, the proper handling of information, and the development of empirical therapy guidelines based on local epidemiology.

[RESUMEN]. Objetivo. Comparar las características epidemiológicas de la resistencia a los antimicrobianos en cepas bacterianas aisladas de muestras de pacientes de servicios hospitalarios y ambulatorios en Ecuador. Métodos. Se realizó un análisis secundario de los datos sobre cepas bacterianas aisladas en muestras de pacientes de servicios hospitalarios y ambulatorios. Se recogieron los datos de la base de datos nacional del 2018 para la vigilancia de la resistencia a los antimicrobianos del Centro de Referencia Nacional para la Resis- tencia a los Antimicrobianos. Las variables incluidas fueron: edad, sexo, entorno hospitalario frente a entorno ambulatorio, tipo de muestra, especies bacterianas detectadas, patrón de resistencia a los antibióticos y zona geográfica. Resultados. En el estudio se incluyeron datos de 57 305 cepas aislamientos bacterianos: 48,8% fueron de pacientes hospitalizados, 55,7% fueron de mujeres y 60,1% fueron de pacientes mayores de 45 años. La orina (42,9%) y la sangre (12,4%) fueron las muestras clínicas más comunes. En general, 77,1% de las cepas bac- terianas aisladas fueron gramnegativas (83% y 71% en pacientes de servicios ambulatorios y hospitalarios, respectivamente). Las especies grampositivas y gramnegativas más comunes fueron Staphylococcus aureus y Escherichia coli, respectivamente. Los niveles de resistencia a los antimicrobianos fueron elevados (hasta 80% en el caso de algunos fármacos antimicrobianos) y fueron más elevados en los pacientes de servicios hospitalarios en comparación con los pacientes de servicios ambulatorios. Se encontró que una variedad de carbapenemasas confiere resistencia a los carbapenémicos (antibióticos de último recurso) en bacterias gramnegativas. Conclusiones. Los resultados del estudio proporcionan una línea de base importante sobre la resistencia a los antimicrobianos en Ecuador, que permitirá el fortalecimiento de las directrices del sistema de vigilancia, la creación de políticas públicas para la estandarización de los métodos de laboratorio, una adecuada gestión de la información y la elaboración de orientaciones de tratamiento empírico basadas en las características epidemiológicas locales.

[RESUMO]. Objetivo. Comparar a epidemiologia da resistência aos antimicrobianos em bactérias isoladas de amostras hospitalares e ambulatoriais no Equador. Métodos. Foi feita uma análise secundária de dados sobre bactérias isoladas de amostras hospitalares e ambulatoriais. Os dados foram obtidos do banco de dados nacional de vigilância da resistência aos antimi- crobianos de 2018 do Centro Nacional de Referência para a Resistência aos Antimicrobianos. As variáveis incluídas foram: idade, sexo, ambiente hospitalar versus ambiente ambulatorial, tipo de espécime, espécies bacterianas identificadas, padrão de resistência a antibióticos e área geográfica. Resultados. Foram incluídos no estudo os dados de 57 305 isolados bacterianos: 48,8% eram de pacientes hospitalizados, 55,7% eram de mulheres e 60,1% eram de pacientes com mais de 45 anos. As amostras clínicas mais comuns foram urina (42,9%) e sangue (12,4%). No total, 77,1% dos isolados bacterianos eram gram-negativos (83% e 71% em pacientes ambulatoriais e pacientes internados, respectivamente). As espécies gram-positivas e gram-negativas mais comuns foram Staphylococcus aureus e Escherichia coli, respectivamente. Os níveis de resistência aos antimicrobianos foram elevados (até 80% para alguns antimi- crobianos) e foram mais elevados em pacientes hospitalizados em comparação com pacientes ambulatoriais. Foram encontradas várias carbapenemases que conferem resistência aos carbapenêmicos (antibióticos de último recurso) em bactérias gram-negativas. Conclusões. Os resultados do estudo fornecem uma importante linha de base sobre a resistência aos anti- microbianos no Equador. Isto permitirá o fortalecimento das diretrizes do sistema de vigilância, a criação de políticas públicas para padronização de metodologias laboratoriais, o manejo adequado de informações e o desenvolvimento de diretrizes para a antibioticoterapia empírica com base na epidemiologia local.

Engineering an inhibitor-resistant human CSF1R variant for microglia replacement.

Hematopoietic stem cell transplantation (HSCT) can replace endogenous microglia with circulation-derived macrophages but has high mortality. To mitigate the risks of HSCT and expand the potential for microglia replacement, we engineered an inhibitor-resistant CSF1R that enables robust microglia replacement. A glycine to alanine substitution at position 795 of human CSF1R (G795A) confers resistance to multiple CSF1R inhibitors, including PLX3397 and PLX5622. Biochemical and cell-based assays show no discernable gain or loss of function. G795A- but not wildtype-CSF1R expressing macrophages efficiently engraft the brain of PLX3397-treated mice and persist after cessation of inhibitor treatment. To gauge translational potential, we CRISPR engineered human-induced pluripotent stem cell-derived microglia (iMG) to express G795A. Xenotransplantation studies demonstrate that G795A-iMG exhibit nearly identical gene expression to wildtype iMG, respond to inflammatory stimuli, and progressively expand in the presence of PLX3397, replacing endogenous microglia to fully occupy the brain. In sum, we engineered a human CSF1R variant that enables nontoxic, cell type, and tissue-specific replacement of microglia.

Nonpharmaceutical interventions reduce the incidence and mortality of COVID-19: A study based on the survey from the International COVID-19 Research Network (ICRN).

The recently emerged novel coronavirus, "severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)," caused a highly contagious disease called coronavirus disease 2019 (COVID-19). It has severely damaged the world's most developed countries and has turned into a major threat for low- and middle-income countries. Since its emergence in late 2019, medical interventions have been substantial, and most countries relied on public health measures collectively known as nonpharmaceutical interventions (NPIs). We aimed to centralize the accumulative knowledge of NPIs against COVID-19 for each country under one worldwide consortium. International COVID-19 Research Network collaborators developed a cross-sectional online survey to assess the implications of NPIs and sanitary supply on the incidence and mortality of COVID-19. The survey was conducted between January 1 and February 1, 2021, and participants from 92 countries/territories completed it. The association between NPIs, sanitation supplies, and incidence and mortality were examined by multivariate regression, with the log-transformed value of population as an offset value. The majority of countries/territories applied several preventive strategies, including social distancing (100.0%), quarantine (100.0%), isolation (98.9%), and school closure (97.8%). Individual-level preventive measures such as personal hygiene (100.0%) and wearing facial masks (94.6% at hospitals; 93.5% at mass transportation; 91.3% in mass gathering facilities) were also frequently applied. Quarantine at a designated place was negatively associated with incidence and mortality compared to home quarantine. Isolation at a designated place was also associated with reduced mortality compared to home isolation. Recommendations to use sanitizer for personal hygiene reduced incidence compared to the recommendation to use soap. Deprivation of masks was associated with increased incidence. Higher incidence and mortality were found in countries/territories with higher economic levels. Mask deprivation was pervasive regardless of economic level. NPIs against COVID-19 such as using sanitizer, quarantine, and isolation can decrease the incidence and mortality of COVID-19.

Epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples, Ecuador, 2018 / Características epidemiológicas de la resistencia antimicrobiana en cepas bacterianas aisladas en pacientes de servicios hospitalarios y ambulatorios de Ecuador, 2018 / Epidemiologia da resistência aos antimicrobianos em bactérias isoladas de amostras hospitalares e ambulatoriais no Equador, 2018

ABSTRACT Objective. To compare the epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples in Ecuador. Methods. A secondary analysis was done of data on bacteria isolated from inpatient and outpatient samples. Data were taken from the 2018 national antimicrobial resistance surveillance database of the National Reference Center for Antimicrobial Resistance. The variables included were: age, sex, inpatient versus outpatient setting, type of specimen, bacterial species identified, pattern of resistance to antibiotics, and geographic area. Results. Data from 57 305 bacterial isolates were included in the study: 48.8% were from hospitalized patients, 55.7% were from women, and 60.1% were from patients older than 45 years. Urine (42.9%) and blood (12.4%) were the most common clinical samples. Overall, 77.1% of bacterial isolates were gram-negative (83% and 71% in outpatients and inpatients, respectively). The most common gram-positive and gram-negative species were Staphylococcus aureus and Escherichia coli, respectively. Antimicrobial resistance levels were high (up to 80% for some antimicrobial drugs), and were higher in hospitalized patients compared with outpatients. A variety of carbapenemases were found to confer resistance to carbapenems (antibiotics of last resort) in gram-negative bacteria. Conclusions. The study findings provide an important baseline on antimicrobial resistance in Ecuador. This will allow the strengthening of guidelines of the surveillance system, the creation of public policies for standardization of laboratory methodologies, the proper handling of information, and the development of empirical therapy guidelines based on local epidemiology.

RESUMEN Objetivo. Comparar las características epidemiológicas de la resistencia a los antimicrobianos en cepas bacterianas aisladas de muestras de pacientes de servicios hospitalarios y ambulatorios en Ecuador. Métodos. Se realizó un análisis secundario de los datos sobre cepas bacterianas aisladas en muestras de pacientes de servicios hospitalarios y ambulatorios. Se recogieron los datos de la base de datos nacional del 2018 para la vigilancia de la resistencia a los antimicrobianos del Centro de Referencia Nacional para la Resistencia a los Antimicrobianos. Las variables incluidas fueron: edad, sexo, entorno hospitalario frente a entorno ambulatorio, tipo de muestra, especies bacterianas detectadas, patrón de resistencia a los antibióticos y zona geográfica. Resultados. En el estudio se incluyeron datos de 57 305 cepas aislamientos bacterianos: 48,8% fueron de pacientes hospitalizados, 55,7% fueron de mujeres y 60,1% fueron de pacientes mayores de 45 años. La orina (42,9%) y la sangre (12,4%) fueron las muestras clínicas más comunes. En general, 77,1% de las cepas bacterianas aisladas fueron gramnegativas (83% y 71% en pacientes de servicios ambulatorios y hospitalarios, respectivamente). Las especies grampositivas y gramnegativas más comunes fueron Staphylococcus aureus y Escherichia coli, respectivamente. Los niveles de resistencia a los antimicrobianos fueron elevados (hasta 80% en el caso de algunos fármacos antimicrobianos) y fueron más elevados en los pacientes de servicios hospitalarios en comparación con los pacientes de servicios ambulatorios. Se encontró que una variedad de carbapenemasas confiere resistencia a los carbapenémicos (antibióticos de último recurso) en bacterias gramnegativas. Conclusiones. Los resultados del estudio proporcionan una línea de base importante sobre la resistencia a los antimicrobianos en Ecuador, que permitirá el fortalecimiento de las directrices del sistema de vigilancia, la creación de políticas públicas para la estandarización de los métodos de laboratorio, una adecuada gestión de la información y la elaboración de orientaciones de tratamiento empírico basadas en las características epidemiológicas locales.

RESUMO Objetivo. Comparar a epidemiologia da resistência aos antimicrobianos em bactérias isoladas de amostras hospitalares e ambulatoriais no Equador. Métodos. Foi feita uma análise secundária de dados sobre bactérias isoladas de amostras hospitalares e ambulatoriais. Os dados foram obtidos do banco de dados nacional de vigilância da resistência aos antimicrobianos de 2018 do Centro Nacional de Referência para a Resistência aos Antimicrobianos. As variáveis incluídas foram: idade, sexo, ambiente hospitalar versus ambiente ambulatorial, tipo de espécime, espécies bacterianas identificadas, padrão de resistência a antibióticos e área geográfica. Resultados. Foram incluídos no estudo os dados de 57 305 isolados bacterianos: 48,8% eram de pacientes hospitalizados, 55,7% eram de mulheres e 60,1% eram de pacientes com mais de 45 anos. As amostras clínicas mais comuns foram urina (42,9%) e sangue (12,4%). No total, 77,1% dos isolados bacterianos eram gram-negativos (83% e 71% em pacientes ambulatoriais e pacientes internados, respectivamente). As espécies gram-positivas e gram-negativas mais comuns foram Staphylococcus aureus e Escherichia coli, respectivamente. Os níveis de resistência aos antimicrobianos foram elevados (até 80% para alguns antimicrobianos) e foram mais elevados em pacientes hospitalizados em comparação com pacientes ambulatoriais. Foram encontradas várias carbapenemases que conferem resistência aos carbapenêmicos (antibióticos de último recurso) em bactérias gram-negativas. Conclusões. Os resultados do estudo fornecem uma importante linha de base sobre a resistência aos antimicrobianos no Equador. Isto permitirá o fortalecimento das diretrizes do sistema de vigilância, a criação de políticas públicas para padronização de metodologias laboratoriais, o manejo adequado de informações e o desenvolvimento de diretrizes para a antibioticoterapia empírica com base na epidemiologia local.

High Resistance to Antibiotics Recommended in Standard Treatment Guidelines in Ghana: A Cross-Sectional Study of Antimicrobial Resistance Patterns in Patients with Urinary Tract Infections between 2017-2021.

Management of urinary tract infections is challenged by increasing antimicrobial resistance (AMR) worldwide. In this study, we describe the trends in antimicrobial resistance of uropathogens isolated from the largest private sector laboratory in Ghana over a five-year period. We reviewed positive urine cultures at the MDS Lancet Laboratories from 2017 to 2021. The proportions of uropathogens with antimicrobial resistance to oral and parenteral antimicrobials recommended by the Ghana standard treatment guidelines were determined. The proportion of multi-drug resistant isolates, ESBL and carbapenemase-producing phenotypes were determined. Of 94,134 urine specimens submitted for culture, 20,010 (22.1%) were culture positive. Enterobacterales was the most common group of organisms, E. coli (70.6%) being the most common isolate and Enterococcus spp. the most common gram-positive (1.3%) organisms. Among oral antimicrobials, the highest resistance was observed to ciprofloxacin (62.3%) and cefuroxime (60.2%) and the least resistance to fosfomycin (1.9%). The least resistance among parenteral antimicrobials was to meropenem (0.3%). The highest multi-drug resistance levels were observed among Klebsiella spp. (68.6%) and E. coli (64.0%). Extended-spectrum beta-lactamase (ESBL) positivity was highest in Klebsiella spp. (58.6%) and E. coli (50.0%). There may be a need to review the Ghana standard treatment guidelines to reflect increased resistance among uropathogens to recommended antimicrobials.

Audit of Clinical Care Received by COVID-19 Patients Treated at a Tertiary Care Hospital of Nepal in 2021.

Like the world over, Nepal was also hard hit by the second wave of COVID-19. We audited the clinical care provided to COVID-19 patients admitted from April to June 2021 in a tertiary care hospital of Nepal. This was a cohort study using routinely collected hospital data. There were 620 patients, and most (458, 74%) had severe illness. The majority (600, 97%) of the patients were eligible for admission as per national guidelines. Laboratory tests helping to predict the outcome of COVID-19, such as D-dimer and C-reactive protein, were missing in about 25% of patients. Nearly all (>95%) patients with severe disease received corticosteroids, anticoagulants and oxygen. The use of remdesivir was low (22%). About 70% of the patients received antibiotics. Hospital exit outcomes of most (>95%) patients with mild and moderate illness were favorable (alive and discharged). Among patients with severe illness, about 25% died and 4% were critically ill, needing further referral. This is the first study from Nepal to audit and document COVID-19 clinical care provision in a tertiary care hospital, thus filling the evidence gap in this area from resource-limited settings. Adherence to admission guidelines was excellent. Laboratory testing, access to essential drugs and data management needs to be improved.

Quality, Equity and Partnerships in Mixed Methods and Qualitative Research during Seven Years of Implementing the Structured Operational Research and Training Initiative in 18 Countries.

Introduction: Qualitative studies are often inadequately reported, making it difficult to judge their appropriateness for decision making in public health. We assessed the publication characteristics and quality of reporting of qualitative and mixed-method studies from the Structured Operational Research and Training Initiative (SORT IT), a global partnership for operational research capacity building. Methods: A cross-sectional analysis of publications to assess the qualitative component using an adapted version of the Consolidated Criteria for Reporting Qualitative Research (COREQ) checklist. Results: In 67 publications involving 18 countries, 32 journals and 13 public health themes, 55 were mixed-methods studies and 12 were qualitative studies. First authorship from low-and-middle-income (LMIC) countries was present in 64 (96%), LMIC last authorship in 55 (82%), and female first authorship in 30 (45%). The mean LMIC institutions represented per publication was five (range 1-11). Sixty-three (94%) publications were open access. Reporting quality was graded as 'good' to 'excellent' in 60 (89%) publications, 'fair' in five (8%) and 'poor' in two (3%). Conclusion: Most SORT IT publications adhered to COREQ standards, while supporting gender equity in authorship and the promotion of LMIC research leadership. SORT IT plays an important role in ensuring quality of evidence for decision making to improve public health.

Trends in Influenza Infections in Three States of India from 2015-2021: Has There Been a Change during COVID-19 Pandemic?

The COVID-19 pandemic and public health response to the pandemic has caused huge setbacks in the management of other infectious diseases. In the present study, we aimed to (i) assess the trends in numbers of samples from patients with influenza-like illness and severe acute respiratory syndrome tested for influenza and the number and proportion of cases detected from 2015−2021 and (ii) examine if there were changes during the COVID-19 period (2020−2021) compared to the pre-COVID-19 period (2015−2019) in three states of India. The median (IQR) number of samples tested per month during the pre-COVID-19 period was 653 (395−1245), compared to 27 (11−98) during the COVID-19 period (p value < 0.001). The median (IQR) number of influenza cases detected per month during the pre-COVID-19 period was 190 (113−372), compared to 29 (27−30) during the COVID-19 period (p value < 0.001). Interrupted time series analysis (adjusting for seasonality and testing charges) confirmed a significant reduction in the total number of samples tested and influenza cases detected during the COVID-19 period. However, there was no change in the influenza positivity rate between pre-COVID-19 (29%) and COVID-19 (30%) period. These findings suggest that COVID-19-related disruptions, poor health-seeking behavior, and overburdened health systems might have led to a reduction in reported influenza cases rather than a true reduction in disease transmission.

Absence of microglia promotes diverse pathologies and early lethality in Alzheimer's disease mice.

Microglia are strongly implicated in the development and progression of Alzheimer's disease (AD), yet their impact on pathology and lifespan remains unclear. Here we utilize a CSF1R hypomorphic mouse to generate a model of AD that genetically lacks microglia. The resulting microglial-deficient mice exhibit a profound shift from parenchymal amyloid plaques to cerebral amyloid angiopathy (CAA), which is accompanied by numerous transcriptional changes, greatly increased brain calcification and hemorrhages, and premature lethality. Remarkably, a single injection of wild-type microglia into adult mice repopulates the microglial niche and prevents each of these pathological changes. Taken together, these results indicate the protective functions of microglia in reducing CAA, blood-brain barrier dysfunction, and brain calcification. To further understand the clinical implications of these findings, human AD tissue and iPSC-microglia were examined, providing evidence that microglia phagocytose calcium crystals, and this process is impaired by loss of the AD risk gene, TREM2.

Immunogenicity of MultiTEP-Platform-Based Recombinant Protein Vaccine, PV-1950R, Targeting Three B-Cell Antigenic Determinants of Pathological α-Synuclein.

Parkinson's disease (PD) and dementia with Lewy bodies (DLB) are characterized by the aberrant accumulation of intracytoplasmic misfolded and aggregated α-synuclein (α-Syn), resulting in neurodegeneration associated with inflammation. The propagation of α-Syn aggregates from cell to cell is implicated in the spreading of pathological α-Syn in the brain and disease progression. We and others demonstrated that antibodies generated after active and passive vaccinations could inhibit the propagation of pathological α-Syn in the extracellular space and prevent/inhibit disease/s in the relevant animal models. We recently tested the immunogenicity and efficacy of four DNA vaccines on the basis of the universal MultiTEP platform technology in the DLB/PD mouse model. The antibodies generated by these vaccines efficiently reduced/inhibited the accumulation of pathological α-Syn in the different brain regions and improved the motor deficit of immunized female mice. The most immunogenic and preclinically effective vaccine, PV-1950D, targeting three B-cell epitopes of pathological α-Syn simultaneously, has been selected for future IND-enabling studies. However, to ensure therapeutically potent concentrations of α-Syn antibodies in the periphery of the vaccinated elderly, we developed a recombinant protein-based MultiTEP vaccine, PV-1950R/A, and tested its immunogenicity in young and aged D-line mice. Antibody responses induced by immunizations with the PV-1950R/A vaccine and its homologous DNA counterpart, PV-1950D, in a mouse model of PD/DLB have been compared.