Activation and Regulation of Blood Vδ2 T Cells Are Amplified by TREM-1+ during Active Pulmonary Tuberculosis.

Triggering receptor expressed on myeloid cells 1 (TREM-1) is a receptor mainly expressed on myeloid cells, and it plays an important role in modulating immune response against infectious agents. The function of TREM-1 on nonmyeloid cells such as Vδ2 T cells has not been characterized, and their role in pulmonary tuberculosis (TB) remains unclear. To assess the expression of TREM-1 on blood Vδ2 T cells from pulmonary TB patients and investigate its mechanism of induction, we exploited flow cytometry analysis to study the expression of TREM-1 on Vδ2 T cells from active pulmonary TB patients and control subjects. In this study we demonstrate that TREM-1 (TREM-1+) is highly expressed on Vδ2 T cells of patients with active pulmonary TB. Unlike TREM-1--expressing Vδ2 T cells, TREM-1+-producing Vδ2 T cells display APC-like phenotypes. Surprisingly, TREM-1+ signaling promotes the Ag-presenting capability of Vδ2 T cells to induce the CD4+ T cell response. TREM-1+Vδ2 T cells induced the proliferation and differentiation of naive CD4+ T cells, as well as the elimination of intracellular mycobacteria. We identified TREM-1+ (but not TREM-1-) as an Ag-presentation amplifier on human blood Vδ2 T cells, and data shed new light on the regulation of Vδ2 T cells in the phase of innate and adaptive immune responses against Mycobacterium tuberculosis infection. Targeting TREM-1+Vδ2 T cells may be a promising approach for TB therapy.

TNF-dependent regulation and activation of innate immune cells are essential for host protection against cerebral tuberculosis.

BACKGROUND: Tuberculosis (TB) affects one third of the global population, and TB of the central nervous system (CNS-TB) is the most severe form of tuberculosis which often associates with high mortality. The pro-inflammatory cytokine tumour necrosis factor (TNF) plays a critical role in the initial and long-term host immune protection against Mycobacterium tuberculosis (M. tuberculosis) which involves the activation of innate immune cells and structure maintenance of granulomas. However, the contribution of TNF, in particular neuron-derived TNF, in the control of cerebral M. tuberculosis infection and its protective immune responses in the CNS were not clear. METHODS: We generated neuron-specific TNF-deficient (NsTNF(-/-)) mice and compared outcomes of disease against TNF(f/f) control and global TNF(-/-) mice. Mycobacterial burden in brains, lungs and spleens were compared, and cerebral pathology and cellular contributions analysed by microscopy and flow cytometry after M. tuberculosis infection. Activation of innate immune cells was measured by flow cytometry and cell function assessed by cytokine and chemokine quantification using enzyme-linked immunosorbent assay (ELISA). RESULTS: Intracerebral M. tuberculosis infection of TNF(-/-) mice rendered animals highly susceptible, accompanied by uncontrolled bacilli replication and eventual mortality. In contrast, NsTNF(-/-) mice were resistant to infection and presented with a phenotype similar to that in TNF(f/f) control mice. Impaired immunity in TNF(-/-) mice was associated with altered cytokine and chemokine synthesis in the brain and characterised by a reduced number of activated innate immune cells. Brain pathology reflected enhanced inflammation dominated by neutrophil influx. CONCLUSION: Our data show that neuron-derived TNF has a limited role in immune responses, but overall TNF production is necessary for protective immunity against CNS-TB.

Neurons are host cells for Mycobacterium tuberculosis.

Mycobacterium tuberculosis infection of the central nervous system is thought to be initiated once the bacilli have breached the blood brain barrier and are phagocytosed, primarily by microglial cells. In this study, the interactions of M. tuberculosis with neurons in vitro and in vivo were investigated. The data obtained demonstrate that neurons can act as host cells for M. tuberculosis. M. tuberculosis bacilli were internalized by murine neuronal cultured cells in a time-dependent manner after exposure, with superior uptake by HT22 cells compared to Neuro-2a cells (17.7% versus 9.8%). Internalization of M. tuberculosis bacilli by human SK-N-SH cultured neurons suggested the clinical relevance of the findings. Moreover, primary murine hippocampus-derived neuronal cultures could similarly internalize M. tuberculosis. Internalized M. tuberculosis bacilli represented a productive infection with retention of bacterial viability and replicative potential, increasing 2- to 4-fold within 48 h. M. tuberculosis bacillus infection of neurons was confirmed in vivo in the brains of C57BL/6 mice after intracerebral challenge. This study, therefore, demonstrates neurons as potential new target cells for M. tuberculosis within the central nervous system.

Assessment of the yellow fever outbreak in Angola from December 2015 through December 2016: A retrospective study.

Background and Aims: The acute tropical infectious disease known as yellow fever (YF) is caused by an arbovirus and is characterized by fever, jaundice, hemorrhage, headache, muscle pain, nausea, vomiting, and fatigue. Angola experienced a yellow fever virus (YFV) outbreak that was documented in December 2015. However, little is known about the outcome of this outbreak. We aimed to demonstrate epidemic features and lessons learned during the YF epidemic in Angola. Methods: A total of 4618 blood samples from suspected YF cases were sent to the Instituto Nacional de Investigação em Saúde (INIS), a national referral and public health laboratory, between December 5, 2015, and December 23, 2016. Sample analyses were conducted using enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) assays. Blood samples were sent from 16 out of the 18 provinces of Angola. Results: We detected 884 (19.1%) cases that were positive for ELISA, which were confirmed by RT-PCR assay. Considering the positive cases, the incidence among male patients was around three times higher (n = 223; 10.9%) than in female patients (n = 59; 2.6%) in the 20-29 age group, followed by the age group 10-19 with n = 211 (6.8%) in males versus n = 108 (3.3%) in females; and the age group 30-39 had n = 68 (4.8%) in males versus n = 28 (1.8%) in females. The other groups had an incidence below 3.0%. The case fatality ratio for YF was in young adults in the age group 20-29 with n = 39 cases, followed by the age group 10-19 with n = 16 cases, and finally the age group 0-9 with n = 13 cases. The other age groups had several deaths by YF below 10 cases. Conclusions: This study demonstrates features of the YF epidemic that occurred in Angola. Also, it demonstrates that YF causes deaths in young people but is preventable by high vaccine coverage. Thus, public health laboratory surveillance must be strengthened to reduce the possibility of emerging and re-emerging human infections.

HIV-1 diversity and pre-treatment drug resistance in the era of integrase inhibitor among newly diagnosed ART-naïve adult patients in Luanda, Angola.

The surveillance of drug resistance in the HIV-1 naïve population remains critical to optimizing the effectiveness of antiretroviral therapy (ART), mainly in the era of integrase strand transfer inhibitor (INSTI) regimens. Currently, there is no data regarding resistance to INSTI in Angola since Dolutegravir-DTG was included in the first-line ART regimen. Herein, we investigated the HIV-1 genetic diversity and pretreatment drug resistance (PDR) profile against nucleoside/tide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors (PIs), and INSTIs, using a next-generation sequencing (NGS) approach with MinION, established to track and survey DRMs in Angola. This was a cross-sectional study comprising 48 newly HIV-diagnosed patients from Luanda, Angola, screened between March 2022 and May 2023. PR, RT, and IN fragments were sequenced for drug resistance and molecular transmission cluster analysis. A total of 45 out of the 48 plasma samples were successfully sequenced. Of these, 10/45 (22.2%) presented PDR to PIs/NRTIs/NNRTIs. Major mutations for NRTIs (2.2%), NNRTIs (20%), PIs (2.2%), and accessory mutations against INSTIs (13.3%) were detected. No major mutations against INSTIs were detected. M41L (2%) and I85V (2%) mutations were detected for NRTI and PI, respectively. K103N (7%), Y181C (7%), and K101E (7%) mutations were frequently observed in NNRTI. The L74M (9%) accessory mutation was frequently observed in the INSTI class. HIV-1 pure subtypes C (33%), F1 (17%), G (15%), A1 (10%), H (6%), and D (4%), CRF01_AG (4%) were observed, while about 10% were recombinant strains. About 31% of detected HIV-1C sequences were in clusters, suggesting small-scale local transmission chains. No major mutations against integrase inhibitors were detected, supporting the continued use of INSTI in the country. Further studies assessing the HIV-1 epidemiology in the era of INSTI-based ART regimens are needed in Angola.

Addressing pandemic-wide systematic errors in the SARS-CoV-2 phylogeny.

The SARS-CoV-2 genome occupies a unique place in infection biology - it is the most highly sequenced genome on earth (making up over 20% of public sequencing datasets) with fine scale information on sampling date and geography, and has been subject to unprecedented intense analysis. As a result, these phylogenetic data are an incredibly valuable resource for science and public health. However, the vast majority of the data was sequenced by tiling amplicons across the full genome, with amplicon schemes that changed over the pandemic as mutations in the viral genome interacted with primer binding sites. In combination with the disparate set of genome assembly workflows and lack of consistent quality control (QC) processes, the current genomes have many systematic errors that have evolved with the virus and amplicon schemes. These errors have significant impacts on the phylogeny, and therefore over the last few years, many thousands of hours of researchers time has been spent in "eyeballing" trees, looking for artefacts, and then patching the tree. Given the huge value of this dataset, we therefore set out to reprocess the complete set of public raw sequence data in a rigorous amplicon-aware manner, and build a cleaner phylogeny. Here we provide a global tree of 3,960,704 samples, built from a consistently assembled set of high quality consensus sequences from all available public data as of March 2023, viewable at https://viridian.taxonium.org. Each genome was constructed using a novel assembly tool called Viridian (https://github.com/iqbal-lab-org/viridian), developed specifically to process amplicon sequence data, eliminating artefactual errors and mask the genome at low quality positions. We provide simulation and empirical validation of the methodology, and quantify the improvement in the phylogeny. Phase 2 of our project will address the fact that the data in the public archives is heavily geographically biased towards the Global North. We therefore have contributed new raw data to ENA/SRA from many countries including Ghana, Thailand, Laos, Sri Lanka, India, Argentina and Singapore. We will incorporate these, along with all public raw data submitted between March 2023 and the current day, into an updated set of assemblies, and phylogeny. We hope the tree, consensus sequences and Viridian will be a valuable resource for researchers.

Immune profile of healthcare professionals six months after vaccination or exposure to SARS-CoV-2 in Angola.

Background and Aims: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is a public health concern. Until 2021, more than 2 million cumulative deaths were reported worldwide. Herein, we investigated the immune profile of healthcare professionals 6 months after vaccination or exposure to SARS-CoV-2 in Angola. Methods: This was a prospective study conducted with 1068 Angolan healthcare professionals between August and December 2021. Participants were screened for the presence of IgG and IgM against SARS-CoV-2. Results: About 9.6% and 98.2% of the participants had prior exposure to SARS-CoV-2 or vaccination against it, respectively. Participants aged between 20 and 40 years (11.2%), female (12.4%), with higher educational level (12.8%), from Luanda (60.3%), and nonhealthcare professionals (8.1%) were the most affected by the SARS-CoV-2. Gender, education, and local residence were related to SARS-CoV-2 exposure (p < 0.05). About 7.3% and 98% of the exposed population developed IgM and IgG after 3 months of exposure, respectively. The AstraZeneca vaccine was the most used, followed by the Jonhson & Johnson and Sputinik. Almost all (98%) participants vaccinated with AstraZeneca had immunity >3 months. Individuals who received only the first dose regardless of the type of vaccine had a higher immunity duration (>3 months) than those who received two doses. For individuals who received the Sputnik and Johnson, the average immunity was lower (<3 months), especially among those who were older (over 40 years old) and exposed to SARS-CoV-2. Conclusion: We observed a high adherence rate to vaccination and a long immunity duration. The immunity duration depended on the type of vaccine. Further studies on the immunity profile in the population exposed to SARS-CoV-2 must be carried out in the general population from Angola to assess antibody-waning periods.

Genomic characterisation of multi-drug resistant Escherichia coli and Klebsiella pneumoniae co-harbouring mcr-1 and mcr-3 genes on a single plasmid from paediatric clinical cases.

OBJECTIVES: Emergence of the plasmid-born mobile colistin resistance (mcr) gene is a growing concern in healthcare. Therefore, this study aimed to genomically characterise multidrug-resistant Escherichia coli and Klebsiella pneumoniae co-harbouring the mcr-1 and mcr-3 genes in young children. METHODS: E. coli (n = 3) and K. pneumoniae (n = 2) were collected from abdominal secretions and blood, respectively. The isolates were screened using tryptone soy broth with 4 µL/mL polymyxin-B. Growing bacteria were identified using the VITEK-2 system, matrix-assisted laser desorption/ionisation time-of-flight, and 16s RNA sequencing, followed by antibiotic susceptibility testing. Metallo-ß-lactamase (MBL) and extended-spectrum ß-lactamase (ESBL) production was also detected. Afterwards, strains were subjected to molecular screening targeting mcr variants and ESBL/MBL-encoding genes. Conjugation, pulsed-field gel electrophoresis, Southern hybridisation, multilocus sequence typing, and phylogenic group detection were performed, along with plasmid-genome sequencing and bioinformatics analysis. RESULTS: E. coli isolates (EC-19-322, 323, and 331) and K. pneumoniae isolates (KP-19-225 and 226) harboured both mcr-1 and mcr-3 genes. These strains were also found to be resistant to more than three classes of antibiotics. The conjugation experiment revealed the presence of mcr-1 and mcr-3 on a single plasmid, and the transmission frequency was 10-2 to 10-3. Both strains were found to be able to produce ESBLs and MBL. E. coli EC-19-322 and 323 were identified as ST131(O25a:H41); SP-19-331, as ST1577 (O16:H30); and K. pneumoniae, as ST231 (K2). All E. coli strains belonged to phylogenetic group B2, and the results of pulsed-field gel electrophoresis supported the multilocus sequence typing findings. CONCLUSION: This study reported the co-occurrence of mcr-1 and mcr-3 genes on a single plasmid in pathogenic ESBL/MBL-producing E. coli and K. pneumoniae isolated from young children.

Demographic characteristics and risk factors related to high blood pressure among healthy blood donors from Luanda, Angola: A retrospective study.

Background and Aims: Hypertension is a public health concern, mainly in resource-limited countries. We investigated the characteristics and risk factors related to high blood pressure in healthy blood donors from, Luanda, the capital city of Angola. Methods: This was a retrospective study that included 343 healthy donors from December 2019 to September 2020. Results: The mean age was 32 ± 9 years. Men represented 93% of the population. Mean systolic blood pressure (SBP) was 131 ± 12.3 mmHg (ranging from 100 to 160 mmHg) and diastolic blood pressure (DBP) was 80.1 ± 9.72 mmHg (from 56.0 to 100 mmHg). DBP was related to age and gender (p < 0.05). About 7.3% of the donors had high-pressure (>140/90 mmHg). Age between 20 and 40 years (odds ratio [OR]: 2.52, p = 0.043), women (OR: 1.87, p = 0.548), nonurbanized areas (OR: 0.39, p = 0.067), high educational level (OR: 0.76, p = 0.637), employed (OR: 0.49, p = 0.491), voluntary donors (OR: 0.87, p = 0.799), blood group B (OR: 2.06, p = 0.346), and Rh- (OR: 0.26, p = 0.104), were potentially related with high-pressure. The high-pressure cases increased from December 2019 (4%) to September 2020 (28%) (p = 0.019). Conclusion: We showed high pressure among the healthy blood donors population. Demographic characteristics, ABO/Rh blood group, and year period are features that should be considered in cardiovascular disease control strategies. Biological and nonbiological features related to blood pressure changes should be considered for further studies in the Angolan population.

Blood pressure pattern among blood donors exposed to SARS-CoV-2 in Luanda, Angola: A retrospective study.

Background and Aims: SARS-CoV-2 infection is a public health concern. Several aspects related to the pattern of infection remain unclear. This study aimed to investigate the blood pressure pattern among blood donors exposed to SARS-CoV-2 in Luanda, Angola, a sub-Saharan African country. Methods: We performed a retrospective analysis containing 343 blood donors from December 2019 to September 2020. Parametric tests compared means while χ 2 and logistic regression checked features associated with high blood pressure and were considered significant when p < 0.05. Results: The mean age of blood donors was 32.2 ± 8.81 years (ranging from 18 to 61 years) and 93% of the men's gender. Overall, 4.7% of the studied population had been exposed to SARS-CoV-2. High blood pressure prevalence increased from unexposed to exposed SARS-CoV-2 (6.7%-18.8%, p = 0.071). SARS-CoV-2 exposure increase systole (131 ± 12.2 mmHg to 136 ± 14.2 mmHg, p = 0.098), diastole (79.9 ± 9.53 mmHg to 84.2 ± 12.7 mmHg, p = 0.086), pulse in beats per minute (72.0 ± 11.1 to 73.7 ± 8.50, p = 0.553), and decrease donating time (6.31 ± 3.72 min to 5.48 ± 1.61 min, p = 0.371). Chances of having high blood pressure were high [OR: 3.20 (95% confidence interval [CI]: 0.85-12.1), p = 0.086] in exposed SARS-CoV-2. Donors exposed to SARS-CoV-2 with abnormal donation time increased from the donor up to 40 years to over 40 years (from 35.7% to 50%, p = 0.696). The mean systolic, diastolic, and pulse pressure were higher for non-O donors (p > 0.05). A significant link was observed, between the Rhesus factor and blood pressure status (p = 0.032). Conclusion: We showed important variations in blood pressure indices of the Angolan population exposed to SARS-CoV-2. Older age and non-O blood groups appear to be important biological factors for SARS-CoV-2 infection, as well as the risk of developing cardiovascular disease after or during SARS-CoV-2 exposure. Further studies assessing the impact on cardiovascular functions with ongoing or long-term SARS-CoV-2 exposure in individuals from resource-limited countries should be considered.

Epidemiology of measles in Angola: Results from routine surveillance from 2015 to 2021.

Background: Measles, an acute infectious disease of extremely contagious viral aetiology, has been eliminated in some parts of the world. To the best of the authors' knowledge, this is the first study on the epidemiological pattern of the measles virus in Angola, and it was carried out through a review of 7 years of observational retrospective data from the national measles laboratory surveillance programme. Methods: A retrospective study using national databases on the laboratory surveillance of measles was performed. Patients of all ages with suspected measles from all provinces of Angola were included. Serum samples were used to detect IgM-type measles-virus-specific antibodies by enzyme-linked immunosorbent assay. Findings: In total, 3690 suspected measles samples were sent to the Instituto Nacional de Investigação em Saúde. There were 962 (26.1%) laboratory-confirmed cases, and the most affected age group was children aged 1-4 years. The highest incidence rate per 100,000 population was found in Benguela (17.9%), followed by Huambo (16.7%) and Cuanza Sul (13.6%). Of the study years, the incidence rate per 1,000,000 population was highest in 2020 (11.9%). The most common complication was diarrhoea (n=406, 42.2%). Of the confirmed cases, 209 (21.7%) were vaccinated, 633 (65.8%) were unvaccinated, and 120 (12.5%) had unknown vaccination status. For all study years, vaccination coverage was <70%. Interpretation: Measles continues to be a serious problem in Angola, and more efforts are needed to increase measles surveillance and achieve a high percentage of vaccination coverage.

Empowering quality data - the Gordian knot of bringing real innovation into healthcare system.

OBJECTIVES: The introduction of Personalised Medicine (PM) into healthcare systems could benefit from a clearer understanding of the distinct national and regional frameworks around the world. Recent engagement by international regulators on maximising the use of real-world evidence (RWE) has highlighted the scope for improving the exploitation of the treasure-trove of health data that is currently largely neglected in many countries. The European Alliance for Personalised Medicine (EAPM) led an international study aimed at identifying the current status of conditions. METHODS: A literature review examined how far such frameworks exist, with a view to identifying conducive factors - and crucial gaps. This extensive review of key factors across 22 countries and 5 regions revealed a wide variety of attitudes, approaches, provisions and conditions, and permitted the construction of a comprehensive overview of the current status of PM. Based on seven key pillars identified from the literature review and expert panels, the data was quantified, and on the basis of further analysis, an index was developed to allow comparison country by country and region by region. RESULTS: The results show that United States of America is leading according to overall outcome whereas Kenya scored the least in the overall outcome. CONCLUSIONS: Still, common approaches exist that could help accelerate take-up of opportunities even in the less prosperous parts of the world.

Framework for Adoption of Next-Generation Sequencing (NGS) Globally in the Oncology Area.

Radical new possibilities of improved treatment of cancer are on offer from an advanced medical technology already demonstrating its significance: next-generation sequencing (NGS). This refined testing provides unprecedentedly precise diagnoses and permits the use of focused and highly personalized treatments. However, across regions globally, many cancer patients will continue to be denied the benefits of NGS as long as some of the yawning gaps in its implementation remain unattended. The challenges at the regional and national levels are linked because putting the solutions into effect is highly dependent on cooperation between regional- and national-level cooperation, which could be hindered by shortfalls in interpretation or understanding. The aim of the paper was to define and explore the necessary conditions for NGS and make recommendations for effective implementation based on extensive exchanges with policy makers and stakeholders. As a result, the European Alliance for Personalised Medicine (EAPM) developed a maturity framework structured around demand-side and supply-side issues to enable interested stakeholders in different countries to self-evaluate according to a common matrix. A questionnaire was designed to identify the current status of NGS implementation, and it was submitted to different experts in different institutions globally. This revealed significant variability in the different aspects of NGS uptake. Within different regions globally, to ensure those conditions are right, this can be improved by linking efforts made at the national level, where patients have needs and where care is delivered, and at the global level, where major policy initiatives in the health field are underway or in preparation, many of which offer direct or indirect pathways for building those conditions. In addition, in a period when consensus is still incomplete and catching up is needed at a political level to ensure rational allocation of resources-even within individual countries-to enable the best ways to make the necessary provisions for NGS, a key recommendation is to examine where closer links between national and regional actions could complement, support, and mutually reinforce efforts to improve the situation for patients.

Insights into SARS-CoV-2 in Angola during the COVID-19 peak: Molecular epidemiology and genome surveillance.

Background: In Angola, COVID-19 cases have been reported in all provinces, resulting in >105,000 cases and >1900 deaths. However, no detailed genomic surveillance into the introduction and spread of the SARS-CoV-2 virus has been conducted in Angola. We aimed to investigate the emergence and epidemic progression during the peak of the COVID-19 pandemic in Angola. Methods: We generated 1210 whole-genome SARS-CoV-2 sequences, contributing West African data to the global context, that were phylogenetically compared against global strains. Virus movement events were inferred using ancestral state reconstruction. Results: The epidemic in Angola was marked by four distinct waves of infection, dominated by 12 virus lineages, including VOCs, VOIs, and the VUM C.16, which was unique to South-Western Africa and circulated for an extended period within the region. Virus exchanges occurred between Angola and its neighboring countries, and strong links with Brazil and Portugal reflected the historical and cultural ties shared between these countries. The first case likely originated from southern Africa. Conclusion: A lack of a robust genome surveillance network and strong dependence on out-of-country sequencing limit real-time data generation to achieve timely disease outbreak responses, which remains of the utmost importance to mitigate future disease outbreaks in Angola.

Use of Honey in the Management of Chemotherapy-Associated Oral Mucositis in Paediatric Patients.

Background: Oral mucositis (OM) is a critical condition during chemotherapy in both adult and child cancer patients. Paediatric cancer patients have a higher prevalence of OM than adult cancer patients. Honey is a natural product that has been reported to have the best tissue healing properties. The present mini-review focused on the evaluation of the effectiveness of oral care with honey products in the treatment and prophylaxis of chemotherapy-induced OM in child patients. Methods: A network of electronic English databases including CINAHL, CENTRAL, EMBASE, MEDLINE and PubMed, were used for primary search from April 2010 to April 2020. We have also considered data collected from ClinicalTrials.gov, Web of Science and Google Scholar. PRISMA software was used to build collective data. Controlled trials were included in this review and were critically appraised by Down and Black. The narrative synthesis was performed. Results: A total number of 346 data of children and adolescents with cancer were considered in this short review. All patients were from three randomized controlled trial articles and two were non-randomised controlled trial articles. Based on the evidence so far revealed, honey may show an effect in the treatment and prophylaxis of OM. The analysis of collected data revealed that the probability value P<0.05. The honey enhanced recovery time and severity of OM were significantly compared with those without honey treatment receiving group of pediatric patients. Conclusion: Honey not only has been shown to have the capability for healing injured tissues but it is also a more economical treatment, and it has fewer side effects compared to synthetic drugs. Honey or honey products can prevent chemotherapy-induced OM (CIOM) and be the best treatment to grade I, II and III CIOM. However, it is disappointing that studies involving children as patients were few, and limited data available so far.

Genetic Characterization of Colistin-Resistant Salmonella enterica ST34 Co-Harbouring Plasmid-Borne mcr-1, bla CTX-M-15 and bla KPC-2 Recovered from a Paediatric Patient in Shenzhen, China.

Background: Since 2015, plasmid-borne mcr-1 has been reported in various bacterial strains in the clinical setting globally. However, the transmission mechanisms of this gene in Salmonella are not well defined. This study aimed to characterize the genomic features of a Salmonella enterica ST34 isolate, which carried a mcr-1, mapped to a carbapenemase and extended spectrum ß-lactamase encoding gene located on the IncX4 plasmid. Methods: Salmonella enterica was recovered from a diarrheal paediatric patient in Shenzhen, China. Antimicrobial susceptibility testing was performed by using the VITEK 2 system. Drug resistance genes were identified using targeted primers and Sanger sequencing. The transferability and genome location of mcr-1 was determined by performing conjugation, S1-PFGE and Southern blot hybridization analysis. WGS was performed by Illumina MiSeq sequencing and was assembled using the A5-Miseq pipeline, and gene annotation was performed using RAST 2.0. The database Centre for Genomic Epidemiology's website was used to identify resistance genes and sequence types (STs). Results: We found that the isolate was extensively drug resistant and belonging to ST34, carrying an IncX4 plasmid with mcr-1, bla KPC-2 and bla CTX-M-15. We also noticed that genes bla PAO, fosA, catB, the mutation in oprD and mexT (MexEF-OprN efflux regulator), and exotoxin-encoding genes (exoS, exoY and exoT) were associated with resistance and virulence in the genome. In addition, heavy metal resistance genes as silP and silE were determined. Conclusion: This study highlights the potential risk of ST34 of Salmonella enterica serotype Typhimurium carrying multiple drug resistance encoding genes in a single IncX4 plasmid.

The Practicality of the Use of Liquid Biopsy in Early Diagnosis and Treatment Monitoring of Oral Cancer in Resource-Limited Settings.

An important driving force for precision and individualized medicine is the provision of tailor-made care for patients on an individual basis, in accordance with best evidence practice. Liquid biopsy(LB) has emerged as a critical tool for the early diagnosis of cancer and for treatment monitoring, but its clinical utility for oral squamous cell carcinoma (OSCC) requires more research and validation. Hence, in this review, we have discussed the current applications of LB and the practicality of its routine use in Africa; the potential advantages of LB over the conventional "gold-standard" of tissue biopsy; and finally, practical considerations were discussed in three parts: pre-analytic, analytic processing, and the statistical quality and postprocessing phases. Although it is imperative to establish clinically validated and standardized working guidelines for various aspects of LB sample collection, processing, and analysis for optimal and reliable use, manpower and technological infrastructures may also be an important factor to consider for the routine clinical application of LB for OSCC. LB is poised as a non-invasive precision tool for personalized oral cancer medicine, particularly for OSCC in Africa, when fully embraced. The promising application of different LB approaches using various downstream analyses such as released circulating tumor cells (CTCs), cell free DNA (cfDNA), microRNA (miRNA), messenger RNA (mRNA), and salivary exosomes were discussed. A better understanding of the diagnostic and therapeutic biomarkers of OSCC, using LB applications, would significantly reduce the cost, provide an opportunity for prompt detection and early treatment, and a method to adequately monitor the effectiveness of the therapy for OSCC, which typically presents with ominous prognosis.

Epidemiological Characteristics and Risk Factors Related to Drug-resistant Tuberculosis in Luanda, Angola.

Tuberculosis (TB) is a major cause of illness and public health concern, especially in resource-limited countries. This study analyzed the characteristics related to anti-TB drug resistance. Moreover, we examined the evidence-based indications for the treatment of active TB in Angola. This study evaluated the medical records of 176 patients screened for TB from January to September 2016 in Luanda, the capital city of Angola. Approximately 66.5% of the patients were newly diagnosed with active TB. The residence area showed a significant relationship with TB (P = 0.025), whereas age group (P = 0.272), gender (P = 0.853), and HIV status (P = 0.284) did not showed any relationship with TB. Overall, 72.4% of TB patients had resistance to at least one of the anti-TB drugs. The risk of anti-TB drug resistance was higher in males (odds ratio [OR]: 1.22; 95% confidence interval [CI]: 0.42-3.58, P = 0.685] and in TB-HIV coinfected patients [OR: 1.39; (95% CI: 0.26-7.28), P = 0.700], whereas it was lower in patients aged 30 years or older (OR: 0.56; 95% CI: 0.18-1.69) P = 0.303) and in patients living in urbanized areas (OR: 0.74; 95% CI: 0.17-3.25; P = 0.685). Our findings showed that drug-resistant TB is emerging in Angola. Further studies on factors related to anti-TB drug resistance are urgently needed to ascertain the magnitude of the problem and to proffer strategies toward TB control in Angola.

Early Evidence of Circulating SARS-CoV-2 in Unvaccinated and Vaccinated Measles Patients, September 2019-February 2020.

BACKGROUND: The global emergence of coronavirus disease 2019 (COVID-19) has challenged healthcare and rapidly spread over the globe. Early detection of new infections is crucial in the control of emerging diseases. Evidence of early recorded COVID-19 cases outside China has been documented in various countries. In this study, we aimed to identify the time of SARS-CoV-2 infection circulation by retrospectively analyzing sera of measles patients, weeks before the reported first COVID-19 cases in Angola. MATERIALS AND METHODS: We examined the humoral response against SARS-CoV-2 by using an enzyme-linked immunosorbent assay (ELISA)-based assay on a combined two-step sandwich enzyme immunoassay method. In total, we received 568 study patients with blood specimens collected from 23 September 2019 to 28 February 2020, 442 sera samples that met the criteria of the study were withdrawn and selected from the overall 568 received samples. In this study, we considered seropositives, patients who tested positive for SARS-CoV-2 immunoglobulin G (IgG) and M (IgM) antibodies with the index value >1. RESULTS: Of the 442 sera samples that met the criteria of the study, 204 were measles seropositive. Forty out of 204 were confirmed reactive to SARS-CoV-2 viral proteins using IgG and IgM more than 2 weeks before the first reported case in Angola. The humoral response analysis showed significant differences (p = 0.01) between the IgG and IgM indexes in the unvaccinated measles patients. Similarly, a significant difference (p = 0.001) was seen between the IgG and IgM indexes in the vaccinated measles patients. CONCLUSION: Here, using the humoral response analysis, we report the identification of early circulation SARS-CoV-2 infection weeks before the first recognized cases in the Republic of Angola.

Evaluation of blood cell count parameters as predictors of treatment failure of malaria in Angola: An observational study.

BACKGROUND: Despite the guidelines provided by the World Health Organization for the treatment of malaria, treatment failure occurs in many hospitalized patients. OBJECTIVE: Evaluate whether blood cell count parameters may serve as predictors for malaria treatment. METHODOLOGY: A cross-sectional study with a quantitative approach. RESULTS: Of the 219 patients, 21.5% showed failure to antimalarial treatment, Patient with 21 and 40 years (72.6%), male (53.4%), from peri-urban area (47.5%), with high parasitemia (59.8%), treated with Arthemeter (90.9%) and the mortality were 5.9%. Significant associations were observed between occupation, level of parasitemia and outcome with resistance to antimalarial treatment (p<0.05). Patients with normal Hb [OR: 0.75 (95% CI: 0.39-1.44), p = 0.393], RBC [OR: 0.83 (95% CI: 0.40-1.72), p = 0.632], RDW [OR: 0.54 (95% CI: 0.27-1.09), p = 0.088], MCV [OR: 0.61 (95% CI: 0.28-1.31), p = 0.204] were less likely to have malaria treatment failures after artemisinin-based therapy failure. In contrast, those with normal values of segmented neutrophils [OR: 0.32 (95% CI: 0.11-0.96), p = 0.042] and lymphocyte counts [OR: 0.24 (95% CI: 0.05-1.04), p = 0.055]. We also found that patients with significant low levels of Hct [OR: 0.31 (95% CI: 0.15-0.64) p = 0.002], and high leukocytes [OR: 8.88 (95% CI: 2.02-37.2), p = 0.004] and normal platelet values [OR: 1.42 (95% CI: 0.73-2.95), p = 0.280] demonstrated high probability of treatment failure. CONCLUSION: The importance of blood cell count parameters in monitoring malaria therapy necessitates the urgent need to re-evaluate Artemether-based therapy. Future studies involving more participants in different settings are needed to provide further evidence.