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Ghana has made significant strides in maternal healthcare under the National Health Insurance Scheme, but more effort is required to achieve the Sustainable Development Goal 3.1 target of A total of 1295 consenting pregnant women participated in the study. Information on sociodemographic characteristics, knowledge about ANC, IPTp and knowledge about malaria were collected by questionnaire. Chi-square tests were used to compare proportions of variables across the two study sites.Higher proportion (70.8%) of participants accessing Battor Catholic Hospital showed adequate knowledge about ANC than 38.1% in Adidome Government Hospital (X2(7) =105.11; pX2(2) =83.37; p80%) possess and use mosquito bed nets for malaria prevention. A lower proportion (46.6%) of participants in Battor Catholic Hospital made the WHO recommended 4-7 visits compared to 50.2% participants in Adidome Government Hospital. Age, marital status, employment and education influenced utilization of antenatal and delivery services. More sensitization on the importance of ANC and hospital delivery is needed in the study areas.

The Noguchi Memorial Institute for Medical Research (NMIMR) is a semi-autonomous institute of the College of Health Sciences, University of Ghana, Legon. Founded in 1979, the Institute was built with a grant aid from the Government of Japan as a gift to the people of Ghana in memory of the renowned Japanese medical scientist, Dr. Hideyo Noguchi, who died from Yellow fever infection while conducting research on the disease in Ghana. The Institute has a three-pronged mandate to conduct health related research, build human capacity and provide specialized diagnostic and disease monitoring services in support of the Ghana Health Service. Over the past 40 years, the Institute has grown to be a leading biomedical research institute in the African region. It has strong and long-standing collaborations with scientists and institutions in Africa, Japan, Europe, Australia and North America on several projects on diseases of public health importance. The Institute also hosts several regional and national centres such as Regional Influenza laboratory. The Institute’s research activities are relevant to the control and prevention of infectious diseases in Ghana, particularly, HIV/AIDS, Tuberculosis, Buruli ulcer, Polio, Malaria and emerging infectious diseases. The Institute also plays a technical/advisory role to government through collaborations with disease control programmes and has since inception provided the country with needed critical evidence in support of health policy as well as laboratory diagnostic services among others. Going forward, the Institute seeks to expand and consolidate its activities in areas of antimicrobial resistance (AMR), clinical trials, genomic surveillance and academic programs and in the next 25 years, NMIMR hopes to approach every research area using the one health approach.

BackgroundSoil Transmitted helminths and schistosomiasis may have implications for pregnancy outcomes especially, in rural communities. In Adidome (a rural community) and Battor (a semi-rural community), soil and water contact activities expose inhabitants to helminth infections. There is, however, limited information on the prevalence and determinants among pregnant women in these areas. The present study was conducted to access the prevalence, knowledge and perceptions about helminthiasis among pregnant women accessing obstetric care at Adidome Government Hospital and Battor Catholic Hospital in the Volta region of Ghana.Methodology/Principal FindingsA cross-sectional survey was conducted by recruiting 1,295 pregnant women out of which only 616 representing 47.5% provided stool samples for analysis. Sociodemographic characteristics and participant knowledge on helminth infection transmission, symptoms and prevention were collected by questionnaire and analyzed using STATA. Preserved stool specimen were processed and examined for helminth eggs by Kato Katz, and genomic DNA extracted from aliquots, was tested forS. mansoniandN. americanus. Prevalence of helminths and participant knowledge were expressed in proportions. Chi-square and Fisher’s exact test were used to show association at P < 0.05 significant level. Intestinal helminth infections found among participants at ANC wereT. trichiura(0.4%),N. americanus(0.4%) andS. mansoni(0.4%). At delivery, a PCR prevalence of 5% was observed forS. mansoni. A high proportion of study participants, 82.5% in Adidome and 87.1% in Battor do not take dewormers on regular basis. Also, a high proportion of participants did not receive any dewormer prior to sample collection. Although knowledge on helminth transmission, risks and prevention were low, levels of prevalence of infection with helminths were also low.Conclusion/SignificanceIntensifying health education as community-based interventions is necessary for the total and effective control and elimination of schistosomiasis and STH in the study area.Author summaryAdidome and Battor are two communities sited close to the Volta Lake which is known to be infested with intermediate hosts for schistosomiasis. Consequently, there has been continued efforts towards control by mass deworming especially among pre-school and school-going children. Despite these control efforts, infections persist as older age groups including pregnant women are often left out of such programs. The fertile soil in the Volta basin encourages farming activities which further expose the people to soil transmitted helminths. There is therefore a need to evaluate community knowledge on helminth infection and precautions to be taken to improve on the control efforts in the areas. The study selected two district Hospitals (a private Hospital in Battor and a government Hospital in Adidome) as study sites. Stool samples collected from pregnant women reporting for the first antenatal care and for delivery were examined for the presence of helminths. Knowledge about helminths among participants were collated by questionnaire. Although prevalences of helminth infections were low, knowledge about helminth infections were also low. However, participants from Battor appear more knowledgeable about helminth infections than those in Adidome. Our study therefore suggests intensive public health education as an intervention for control.

Sterile protection against clinical malaria has been achieved in animal models and experimental human challenge studies involving immunization with radiation attenuated Plasmodium falciparum sporozoite vaccines as well as by live sporozoites under chloroquine prophylaxis. Parasite-specific IFN-γ and granzyme B-secreting CD8 + T cells have been identified as key mediators of protection. Although the exact parasite targets of protective CD8 + T cell responses are not fully defined, responses against a handful of vaccine candidate antigens have been associated with protection. Identifying the T cell targets in these antigens will facilitate the development of simpler, cost-effective, and efficacious next generation multi-epitope vaccines. The aim of this study was to identify immunodominant portions of four malaria vaccine candidate antigens using peripheral blood mononuclear cells (PBMCs) from adults with life-long exposure to malaria parasites. Cryopreserved PBMCs from 291 HLA-typed subjects were stimulated with pools of overlapping 15mer peptides spanning the entire sequences of P. falciparum circumsporozoite protein (CSP, 9 pools), apical membrane antigen 1 (AMA1, 12 pools), thrombospondin related anonymous protein (TRAP, 6 pools) and cell traversal for ookinetes and sporozoites (CelTOS, 4 pools) in FluoroSpot assays. 125 of 291 subjects made IFN-γ responses to 30 of the 31 peptide pools tested and 22 of 291 made granzyme B responses, with 20 making dual responses. The most frequent responses were to the CSP C-terminal region and the least frequent responses were to TRAP and CelTOS. There was no association between FluoroSpot responses and active malaria infection, detected by either microscopy, RDT, or PCR. In conclusion, CSP and AMA1 have relatively higher numbers of epitopes that trigger IFN-γ and granzyme B-secreting T cells in adults with life-long malaria parasite exposure compared to the other two antigens tested, and highlights the continued relevance of these two antigens as vaccine candidates.

BackgroundWith the decline in malaria transmission due to global efforts, a more sensitive tool is needed to monitor transmission intensity and pattern at the micro-level. Though transmission in a broader area may be similar, factors such as sanitation, practices of open water storage, early morning and evening activities, outdoor sleeping and agricultural practices within communities could cause differences in exposure and thus transmission. This study thus probed malaria transmission at a micro-level using serology in the Hohoe Municipality of Ghana.MethodsThis cross-sectional study involved 327 asymptomatic children aged 1-12 years in both rural (196) and urban (131) communities in the Hohoe municipality. Total IgG responses specific for three P. falciparum antigens (CSP, MSP2-FC27, MSP2-3D7) were determined in plasma eluted from dried blood spots using indirect ELISA.ResultsA higher proportion of individuals in the rural area had parasites by both microscopy and PCR. Total IgG levels and seroprevalence were higher in rural compared to urban communities (pConclusionThe results suggest that though the district is categorized as having medium malaria transmission, differences within settlements may influence malaria transmission reflecting in antibody levels and prevalence of malaria antigen-specific IgG.

Background: Plant as a source of medicine has gained international popularity in recent times because of its natural origin, availability in local communities, cheaper to purchase, ease of administration, and its usefulness as an alternative treatment in case of numerous side effects and drug resistance. However, the use of herbal formulations can also result in short-term and long-term organ damage or dysfunction to the host. In this study, chloroform fractions of the leaves of two medicinal plants, Alchornea cordifolia (ACL) and Carapa procera (CPL), were investigated for their toxicological and anti-malarial effects in murine models.Method: Acute (14-day) and sub-acute (28-day) studies were conducted based on the Organization for Economic Cooperation and Development (OECD) Guidelines in Institute for Cancer Research (ICR) mice and Sprague Dawley (SD) rats respectively. A dosage of 2000 mg/kg body weight was administered orally to each ICR mouse during the acute study and 100, 300, and 1000 mg/kg body weight to each SD rat during the sub-acute study. A 5-day curative anti-plasmodial activity was assessed in ICR mouse model.Results: The assessment of toxicity revealed that all three fractions did not influence mortality, clinical appearance, body weight gain, or necropsy at the various doses. Hematological and serum biochemical analysis indicated no significant elevations in liver and renal function parameters. Histopathological examinations of the liver indicated reversible liver degeneration with the chloroform fraction of the 100% ethanol extract of Carapa procera leaves (CPL100%) at 1000 mg/kg. Anti-plasmodial assessments showed CPL100% exhibiting dose-dependent anti-plasmodial activity from 16% to 26.67%. On the other hand, chloroform fraction of the 100% ethanol extract of Alchornea cordifolia leaves (ACL100%) showed declining anti-plasmodial activity from 21.1% to 15.1%.Conclusion: These preliminary findings demonstrate that chloroform fractions of the leaves of Carapa procera and Alchornea cordifolia may be safe agents for treating malaria hence further development for drug discovery must be pursued.

The coronavirus disease 2019 (COVID-19) pandemic devastated countries worldwide, and resulted in a global shutdown. Not all infections are symptomatic and hence the extent of SARS-CoV-2 infection in the community is unknown. The paper presents the dynamics of the SARS-CoV-2 epidemic in the Greater Accra Metropolis, describing the evolution of seroprevalence through time and by age group. Three repeated independent population-based surveys at 6-week intervals were conducted in from November 2020 to July 2021. The global and by age-groups weighted seroprevalences were estimated and the risk factors for SARS-CoV-2 antibody seropositivity were assessed using logistic regression. The overall age-standardized SARS-CoV-2 antibody seroprevalence for both spike and nucleocapsid increased from 13.8% (95% CI 11.9, 16.1) in November 2020 to 39.6% (95% CI 34.8, 44.6) in July 2021. After controlling for gender, marital status, education level, and occupation, the older age group over 40 years had a higher odds of seropositivity than the younger age group (OR 3.0 [95% CI 1.1–8.5]) in the final survey. Pupils or students had 3.3-fold increased odds of seropositivity (OR 3.2 [95% CI 1.1–8.5]) compared to the unemployed. This study reinforces that, SARS-CoV-2 infections have been significantly higher than reported.

Background: In Ghana, Hepatitis B virus (HBV) infection remains a major public health threat as in many parts of the world. Even with an effective vaccine, there are shortfalls with low vaccine coverage among adults. To create awareness and encourage vaccination, community engagement and public-private partnerships are needed in endemic settings to help fund campaigns and offer vaccinations at no cost to under privileged people. Methods: Participants from the University of Ghana community and its immediate environs were registered, taken through pre-counselling sessions where they were educated on hepatitis transmission and prevention before consenting. Eligible participants were screened for HBV markers with rapid test kits. All HBsAb-negative participants were recommended for initial vaccination at the event, whilst the subsequent shots were administered. Hepatitis B surface Antigen-positive participants were counselled and referred for appropriate care. Results: A total of 297 people, comprising of 126 (42%) males and 171 (58%) females aged between 17 and 67 years were screened. Amongst these, 246 (82.8%) showed no detectable protective antibodies against HBV and all of them agreed to and were given the first dose HBV vaccine. Additionally, 19 (6.4%) individuals tested positive for HBsAg and were counselled and referred to specialists for further assessment and management. We found that 59 (19.9%) of our participants had previously initiated HBV vaccination and had taken at least one dose of the vaccine more than 6 months prior to this screening, 3 of whom tested positive for HBsAg. For the three-dose HBV vaccines deployed, a little over 20% (50/246) and a further 17% (33/196) did not return for the second and the third doses respectively, resulting in an overall 66% (163/246) of persons who completed all three vaccinations. Conclusions: A key lesson from this exercise is the need for partnerships between the health sector and corporate bodies to increase and improve awareness, care, and management of hepatitis. Additionally, home and school vaccination programmes may be adopted to enhance vaccine uptake and adherence to the vaccination schedule. We plan to extend this screening exercise to deprived and/or rural communities where HBV incidence may be higher than in urban communities.

BackgroundThe human host elicits specific immune responses after exposure to various life stages of the malaria parasite as well as components of mosquito saliva injected into the host during a mosquito bite. This study describes differences in IgG responses against antigens derived from the sporozoite (PfCSP), asexual stage parasite (PfEBA175) and the gametocyte (Pfs230) in addition to anAnopheles gambiaesalivary gland antigen (gSG6-P1) in two communities in Ghana with similar blood stage malaria parasite prevalence.MethodologyThis study used archived plasma samples collected from an earlier cross-sectional study that enrolled volunteers aged from 6 months to 70 years from Simiw, peri-urban community (N=347) and Obom, rural community (N=291). An archived thick and thin blood smear for microscopy was used for the estimation ofPlasmodiumparasite density and species and DNA extraction from blood spots andP. falciparumconfirmation was performed using PCR. This study used the stored plasma samples to determine IgG antibody levels toPlasmodium falciparumandAnophelessalivary antigens using indirect ELISA.ResultsIndividuals from Simiw had significantly higher levels of IgG against mosquito gSG6-P1 (median (95%CI)) (2.590 (2.452-2.783) ng/mL) compared to those from Obom (2.119 (1.957-2.345) ng/mL), p0.05).ConclusionIn conclusion, malaria transmission dynamics is highly complex. The similar malaria transmission intensity identified in the two communities resulted from a different combination of vector, environmental and host factors. With one community likely having a higher prevalence of uninfected mosquitoes and the other a larger reservoir of gametocyte carriers.

Cerebral malaria (CM) may cause death or long-term neurological damage in children, and several host genetic risk factors have been reported. Malaria-specific immunoglobulin (Ig) G3 antibodies are crucial to human immune response against malaria. The hinge region of IgG3 exhibits length polymorphism (with long [L], medium [M], and short [S] alleles), which may influence its functionality. We studied IgG3 hinge region length polymorphisms in 136 Ghanaian children with malaria. Using logistic regression models, we found that children with the recessive MM allotype encoding medium IgG3 hinge region length had an increased risk of CM (adjusted odds ratio, 6.67 [95% confidence interval,1.30–34.32]; P=.004) . This has implications for future epidemiological studies on CM.

Background Malaria eradication requires a combined effort involving all available control tools, and these efforts would be complemented by an effective vaccine. The antigen targets of immune responses may show polymorphisms that can undermine their recognition by immune effectors and hence render vaccines based on antigens from a single parasite variant ineffective against other variants. This study compared the influence of allelic polymorphisms in Plasmodium falciparum apical membrane antigen 1 (PfAMA1) peptide sequences from three strains of P. falciparum (3D7, 7G8 and FVO) on their function as immunodominant targets of T cell responses in high and low malaria transmission communities in Ghana. Methods Peripheral blood mononuclear cells (PBMCs) from 10 subjects from a high transmission area (Obom) and 10 subjects from a low transmission area (Legon) were tested against 15 predicted CD8 + T cell minimal epitopes within the PfAMA1 antigen of multiple parasite strains using IFN-γ ELISpot assay. The peptides were also tested in similar assays against CD8 + enriched PBMC fractions from the same subjects in an effort to characterize the responding T cell subsets. Results In assays using unfractionated PBMCs, two subjects from the high transmission area, Obom, responded positively to four (26.7%) of the 15 tested peptides. None of the Legon subject PBMCs yielded positive peptide responses using unfractionated PBMCs. In assays with CD8 + enriched PBMCs, three subjects from Obom made positive recall responses to six (40%) of the 15 tested peptides, while only one subject from Legon made a positive recall response to a single peptide. Overall, 5 of the 20 study subjects who had positive peptide-specific IFN-γ recall responses were from the high transmission area, Obom. Furthermore, while subjects from Obom responded to peptides in PfAMA1 from multiple parasite strains, one subject from Legon responded to a peptide from 3D7 strain only. Conclusions The current data demonstrate the possibility of a real effect of PfAMA1 polymorphisms on the induction of T cell responses in malaria exposed subjects, and this effect may be more pronounced in communities with higher parasite exposure.

The changes occurring in the T cell repertoire during clinical malaria infection in children remain unknown. In this study, we undertook the first detailed comparative study of the T cell repertoire in African children with and without clinical malaria to test the hypothesis that clonotypic expansions that occur during P. falciparum infection will contribute to the generation of a T cell repertoire that is unique to each disease state. We profiled the complementarity-determining region 3 (CDR3) of the TCRβ chain sequences from children with Plasmodium falciparum infections (asymptomatic, uncomplicated and severe malaria) and compared these with sequences from healthy children. Interestingly, we discovered that children with symptomatic malaria have a lower TCR diversity and frequency of shared (or “public”) TCR sequences compared to asymptomatic children. Also, TCR diversity was inversely associated with parasitemia. Furthermore, by clustering TCR sequences based on their predicted antigen specificities, we identified a specificity cluster, with a 4-mer amino acid motif, that is overrepresented in the asymptomatic group compared to the diseased groups. Further investigations into this finding may help in delineating important antigenic targets for vaccine and therapeutic development. The results show that the T cell repertoire in children is altered during malaria, suggesting that exposure to P. falciparum antigens disrupts the adaptive immune response, which is an underlying feature of the disease.

BackgroundVaccine-preventable diseases (VPDs) persist globally with a disproportionately high burden in Low and Middle-Income Countries (LMICs). Although this might be partly due to the failure to sustain vaccination coverage above 90% in some WHO regions, a more nuanced understanding of VPD transmission beyond vaccination coverage may unveil other important factors in VPD transmission and control. This study identified VPDs hotspots and explored their relationships with ecology, urbanicity and land-use variations (Artisanal and Small-scale Gold Mining (ASGM) activities) in Ghana.MethodsDistrict-level disease count data from 2010 to 2014 from the Ghana Health Service (GHS) and population data from the Ghana Population and Housing Census (PHC) were used to determine clustering patterns of six VPDs (Measles, Meningitis, Mumps, Otitis media, Pneumonia and Tetanus). Spatial and space-time cluster analyses were implemented in SaTScan using the discrete Poisson model.P-values were estimated using a combination of sequential Monte Carlo, standard Monte Carlo, and Gumbel approximations.ResultsThe study found a preponderance for VPD hotspots in the northern parts of Ghana and northernmost ecological zones (Sudan Savannah and Guinea Savannah). Incidence of meningitis was higher in the Sudan Savannah ecological zone relative to: Tropical Rain Forest (p = 0.001); Semi Deciduous Forest (p p = 0.045), incidence of the other five VPDs did not differ across the urban-rural divide. Whereas spatial analysis suggested that some VPD hotspots (tetanus and otitis media) occur more frequently in mining districts in the southern part of the country, a Mann-Whitney U test revealed a higher incidence of meningitis in non-mining districts (p = 0.019). Pneumonia and meningitis recorded the highest (722.8 per 100,000) and least (0.8 per 100,000) incidence rates respectively during the study period.ConclusionThis study shows a preponderance of VPD hotspots in the northern parts of Ghana and in semi-arid ecoclimates. The relationship between ASGM activities and VPD transmission in Ghana remains blurred and requires further studies with better spatial resolution to clarify.

Background and aim Most developing countries resort to medicinal plants for treating diseases, but few of these have scientific backing for their use. The aim of the study was to validate traditional use of Morinda lucida leaves in treating inflammation and determine the mechanism of action. Experimental procedure Effect of hydroethanolic leaf extract of M. lucida (HEML) on localized inflammation was evaluated using rat paw edema presented by sub-planter injections of λ-carrageenan, histamine or serotonin in separate experiments. Systemic inflammation was evaluated by lipopolysaccharide (LPS)-induced hyperthermia. Antioxidant activity of HEML was also evaluated using the free-radical scavenging assay. Results and conclusion No mortalities were recorded in acute toxicity assay after administering 5000 mg/kg HEML to rats. It showed very good activity against localized and systemic inflammation in inverse dose-dependent manner and caused reduction in nitric oxide and prostaglandin E−2 levels by affecting expression of inducible nitric oxide synthase, but not cyclooxygenases-2 in LPS-activated RAW 264.7 murine macrophages. HEML reduced pro-inflammatory cytokines interleukin (IL)-1β and tumor necrotic factor, but elevated levels of anti-inflammatory cytokine IL-10 in vitro. HEML contains saponins, reducing sugars, polyphenols and flavonoids and showed antioxidant activity with EC50 = 0.6415 ± 0.0027 mg/ml. In conclusion, this study provides evidence that HEML possesses anti-inflammatory activity, possibly through modulation of production of early/late phase inflammation mediators.

Background Malaria related mortality is associated with significant deregulation of host inflammatory factors such as interferon-inducible protein 10, a member of the CXC or α-subfamily (CXCL10), and host angiogenic factors such as angiopoietin 1 (Ang-1) and angiopoietin 2 (Ang-2). However, detection of these factors in malaria patients requires the drawing of blood, which is invasive and increases the risk of accidental blood-borne infections. There has been an increased interest in the use of saliva as the body fluid of choice for the diagnosis of many infectious diseases including malaria. Here, saliva levels of CXCL10, Ang-1, and Ang-2 previously shown to be predictive of severe malaria in malaria patients in Ghana were assessed in malaria patients. Methods This study was conducted in the Shai-Osudoku District Hospital in Dodowa, Accra, Ghana and the study population comprised 119 malaria patients and 94 non-malaria subjects. The non-malaria subjects are healthy community participants with no malaria infection. Plasma and saliva levels of CXCL10, Ang-1 and Ang-2 of the study participants were measured using an enzyme-linked immunoassay. Complete blood counts of each participant were measured with a haematology autoanalyzer. Pearson correlation was used to evaluate the correlation between plasma and saliva levels of each biomarker in malaria patients. A p-value of Results The non-malaria subjects had a median age of 29 years compared to 23 years for malaria patients (p = 0.001). Among the malaria patients, there was a strong significant relationship between CXCL10 (R2 = 0.7, p 2 = 0.7, p Conclusions This study provides the first evidence of elevated levels of CXCL10 and Ang-2 in the saliva of malaria patients. Detection of CXCL10, Ang-1 and Ang-2 in saliva may have a potential application for non-invasive malaria diagnosis.

Nearly 60% of Plasmodium falciparum proteins are still uncharacterized and their functions are unknown. In this report, we carried out the functional characterization of a 45 kDa protein (PF3D7_1459400) and showed its potential as a target for blood stage malaria vaccine development. Analysis of protein subcellular localization, native protein expression profile, and erythrocyte invasion inhibition of both clinical and laboratory parasite strains by peptide antibodies suggest a functional role of PF3D7_1459400 protein during erythrocyte invasion. Also, immunoreactivity screens using synthetic peptides of the protein showed that adults resident in malaria endemic regions in Ghana have naturally acquired plasma antibodies against PF3D7_1459400 protein. Altogether, this study presents PF3D7_1459400 protein as a potential target for the development of peptide-based vaccine for blood-stage malaria. Impact statement Plasmodium falciparum malaria is a global health problem. Erythrocyte invasion by P. falciparum merozoites appears to be a promising target to curb malaria. We have identified and characterized a novel protein that is involved in erythrocyte invasion. Our data on protein subcellular localization, stage-specific protein expression pattern, and merozoite invasion inhibition by α-peptide antibodies suggest a role for PF3D7_1459400 protein during P. falciparum erythrocyte invasion. Even more, the human immunoepidemiology data present PF3D7_1459400 protein as an immunogenic antigen which could be further exploited for the development of new anti-infective therapy against malaria.

Plasmodium falciparum erythrocyte invasion phenotyping assays are a very useful tool for assessing parasite diversity and virulence, and for characterizing the formation of ligand–receptor interactions. However, such assays need to be highly sensitive and reproducible, and the selection of labeling dyes for differentiating donor and acceptor erythrocytes is a critical factor. We investigated the suitability of cell trace far-red (CTFR) as a dye for P. falciparum invasion phenotyping assays. Using the dyes carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and dichloro dimethyl acridin one succinimidyl ester (DDAO-SE) as comparators, we used a dye-dilution approach to assess the limitations and specific staining procedures for the applicability of CTFR in P. falciparum invasion phenotyping assays. Our data show that CTFR effectively labels acceptor erythrocytes and provides a stable fluorescent intensity at relatively low concentrations. CTFR also yielded a higher fluorescence intensity relative to DDAO-SE and with a more stable fluorescence intensity over time. Furthermore, CTFR did not affect merozoites invasion of erythrocytes and was not toxic to the parasite’s intraerythrocytic development. Additionally, CTFR offers flexibility in the choice of combinations with several other DNA dyes, which broaden its usage for P. falciparum erythrocyte invasion assays, considering a wider range of flow cytometers with various laser settings. Impact statement In recent years, flow cytometry has become a cornerstone in investigating P. falciparum phenotypic diversity using multiple dyes to discriminate between donor and acceptor erythrocytes. To broaden the applicability of such assays, we optimized the staining conditions of a newly developed cytoplasmic dye, cell trace far-red (CTFR), and assessed its suitability for use in P. falciparum invasion phenotyping assays. We showed that CTFR has a very narrow emission peak excited by red lasers. Furthermore, CTFR labeling of target erythrocytes, achieved even at low concentrations, is stable over time and did not impair parasite development. P. falciparum erythrocyte invasion phenotyping assays revealed that CTFR is suitable for use in combination with several DNA dyes in multiplex assays. This will allow for high throughput phenotyping of parasites as well as facilitate the evaluation of preference of erythrocytes by merozoites. Altogether, these make screening for potential invasion-blocking interventions possible.

The RTS,S/AS01E vaccine targets the circumsporozoite protein (CSP) of the Plasmodium falciparum (P. falciparum) parasite. Protein microarrays were used to measure levels of IgG against 1000 P. falciparum antigens in 2138 infants (age 6-12 weeks) and children (age 5-17 months) from 6 African sites of the phase III trial, sampled before and at 4 longitudinal visits after vaccination. One month postvaccination, IgG responses to 17% of all probed antigens showed differences between RTS,S/AS01E and comparator vaccination groups, whereas no prevaccination differences were found. A small subset of antigens presented IgG levels reaching 4- to 8-fold increases in the RTS,S/AS01E group, comparable in magnitude to anti-CSP IgG levels (~11-fold increase). They were strongly cross-correlated and correlated with anti-CSP levels, waning similarly over time and reincreasing with the booster dose. Such an intriguing phenomenon may be due to cross-reactivity of anti-CSP antibodies with these antigens. RTS,S/AS01E vaccinees with strong off-target IgG responses had an estimated lower clinical malaria incidence after adjusting for age group, site, and postvaccination anti-CSP levels. RTS,S/AS01E-induced IgG may bind strongly not only to CSP, but also to unrelated malaria antigens, and this seems to either confer, or at least be a marker of, increased protection from clinical malaria.

The RTS,S/AS01E vaccine targets the circumsporozoite protein (CSP) of the Plasmodium falciparum parasite. Using protein microarrays, levels of IgG to 1,000 P. falciparum antigens were measured in 2,138 infants (age 6-12 weeks) and children (age 5-17 months) from 6 African sites of the phase 3 trial, sampled before and at four longitudinal visits after vaccination. One month post-vaccination, IgG responses to 17% of all probed antigens showed differences between RTS,S/AS01E and comparator vaccination groups, whereas no pre-vaccination differences were found. A small subset of antigens presented IgG levels reaching 4- to 8-fold increases in the RTS,S/AS01E group, comparable in magnitude to anti-CSP IgG levels (∼11-fold increase). They were strongly cross-correlated and correlated with anti-CSP levels, waning similarly over time and re-increasing with the booster dose. Such an intriguing phenomenon may be due to cross-reactivity of anti-CSP antibodies with these antigens. RTS,S/AS01E vaccinees with strong off-target IgG responses had an estimated lower clinical malaria incidence after adjusting for age group, site and post-vaccination anti-CSP levels. RTS,S/AS01E-induced IgG may bind strongly not only to CSP, but to unrelated malaria antigens, and this seems to either confer, or at least be a marker of, increased protection from clinical malaria.

Background In malaria, clinical disease has been associated with increased levels of endothelial activation due to the sequestration of infected erythrocytes. However, the levels and impact of endothelial activation and pro-angiogenic molecules such as vascular endothelial growth factor (VEGF)–A and its receptor vascular endothelial growth factor receptor 2 (VEGFR2) in asymptomatic malaria have not been well characterized. Methods Blood samples were obtained from community children for malaria diagnosis using microscopy and polymerase chain reaction. A multiplex immunoassay was used to determine the levels of intracellular adhesion molecule (ICAM)–1, vascular endothelial growth factor (VEGF)–A, and VEGFR2 in the plasma of children with microscopic or submicroscopic asymptomatic parasitemia and compared with levels in uninfected controls. Results Levels of ICAM-1, VEGF-A, and VEGFR2 were significantly increased in children with microscopic asymptomatic parasitemia compared with uninfected controls. Also, levels of VEGF-A were found to be inversely associated with age. Additionally, a receiver operating characteristic analysis revealed that plasma levels of ICAM-1 (area under the curve [AUC], 0.72) showed a moderate potential in discriminating between children with microscopic malaria from uninfected controls when compared with VEGF-A (AUC, 0.67) and VEGFR2 (AUC, 0.69). Conclusions These data imply that endothelial activation and pro-angiogenic growth factors could be one of the early host responders during microscopic asymptomatic malaria and may play a significant role in disease pathogenesis.

The current global malaria control and elimination agenda requires development of additional effective disease intervention tools. Discovery and characterization of relevant parasite antigens is important for the development of new diagnostics and transmission monitoring tools and for subunit vaccine development. This study assessed the natural antibody response profile of seven novel Plasmodium falciparum pre-erythrocytic antigens and their potential association with protection against clinical malaria. Antigen-specific antibody levels in plasma collected at six time points from a longitudinal cohort of one-to-five year old children resident in a seasonal malaria transmission area of northern Ghana were assessed by ELISA. Antibody levels were compared between parasite-positive and parasite-negative individuals and the association of antibody levels with malaria risk assessed using a regression model. Plasma antibody levels against five of the seven antigens were significantly higher in parasite-positive children compared to parasite-negative children, especially during low transmission periods. None of the antigen-specific antibodies showed an association with protection against clinical malaria. The study identified five of the seven antigens as markers of exposure to malaria, and these will have relevance for the development of disease diagnostic and monitoring tools. The vaccine potential of these antigens requires further assessment.

BackgroundDiverse signalling pathways are involved in carcinogenesis and one of such pathways implicated in many cancers is the interleukin 6/signal transducer and activator of transcription 3 (IL-6/STAT3) signalling pathway. Therefore, inhibition of this pathway is targeted as an anti-cancer intervention. This study aimed to establish the effect of cryptolepine, which is the main bioactive alkaloid in the medicinal plantCryptolepis sanguinolenta, on the IL-6/STAT3 signalling pathway.MethodsFirst, the effect of cryptolepine on the IL-6/STAT3 pathway in human hepatoma cells (HepG2 cells) was screened using the Cignal Finder Multi-Pathway Reporter Array. Next, to confirm the effect of cryptolepine on the IL-6/STAT3 signalling pathway, the pathway was activated using 200 ng/mL IL-6 in the presence of 0.5–2 μM cryptolepine. The levels of total STAT3, p-STAT3 and IL-23 were assessed by ELISA.ResultsCryptolepine downregulated 12 signalling pathways including the IL-6/STAT3 signalling pathway and upregulated 17 signalling pathways. Cryptolepine, in the presence of IL-6, decreased the levels of p-STAT3 and IL-23 in a dose-dependent fashion.ConclusionOur results demonstrated that cryptolepine inhibits the IL-6/STAT3 signalling pathway, and therefore cryptolepine-based remedies such asCryptolepis sanguinolentacould potentially be used as an effective immunotherapeutic agent for hepatocellular carcinoma and other cancers.

The rapid development in Morocco with an urban and agricultural expansion is putting pressure on ecosystem services. An integrated assessment of these services at different spatial and temporal scales will ensure their sustainability. This paper used Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST) to quantify and map multiple ecosystem services in Morocco from 1992 to 2015. The results show that urban areas, agriculture, sparse vegetation, and forests increased by 93%, 2%, 8%, and 9% whereas grassland and bare areas decreased by 80% and 2% respectively. This caused an increase in carbon storage by 1.7% and decreases in water yield, sediment export, and nitrogen and phosphorus exports by 15.5%, 7.4%, and 0.4% respectively. The overall ecosystem services of Morocco showed high values in the north but lower values in the east and south. Land management activities which were implemented in a selected watershed in the east using the Resource Investment Optimization System (RIOS) caused carbon storage to further increase by 0.2% whereas water yield, sediment export, nitrogen export, and phosphorus export declined by 0.07%, 0.09%, 0.17%, and 0.09%. Ecosystem services could be improved at both national and watershed scales when management activities such as reforestation and pasture management are implemented at specific watersheds.

Detection of antibody reactivity to appropriate, specific parasite antigens may constitute a sensitive and cost-effective alternative to current tools to monitor malaria transmission across different endemicity settings. This study aimed to determine the suitability of IgG responses to a number of P. falciparum antigens as markers of transmission intensity and pattern. Antibody responses to multiple malaria antigens were determined in 905 participants aged 1–12 years from three districts with low (Keta), medium (Hohoe) and high (Krachi) transmission intensity in the Volta region of Ghana. Blood film microscopy slides and dry blood spots (DBS) were obtained for parasitaemia detection and antibody measurement, respectively. Sera were eluted from DBS and levels of IgG specific for 10 malaria antigens determined by a multiplex assay. Results were compared within and among the districts. Total IgG responses to MSPDBL1, MSPDBLLeucine, MSP2-FC27, RAMA, and PfRh2a and PfRh2b were higher in Krachi than in Hohoe and Keta. Seroprevalence of IgG specific for MSPDBLLeucine, RON4, and PfRh2b were also highest in Krachi. Responses to RALP-1, PfRh2a and PfRh2b were associated with patent but asymptomatic parasitaemia in Keta, while responses to MSPDBL1, MSPDBLLeucine, MSP2-FC27, RAMA, Rh2-2030, and PfRh2b were associated with parasite carriage in Hohoe, but not in Krachi. Using ROC analysis, only PfRh2b was found to predict patent, but asymptomatic, parasitaemia in Keta and Hohoe. Antibody breadth correlated positively with age (r = 0.29, pPfRh2b has potential as a marker of malaria transmission intensity and pattern. This could have implications for malaria control programs and vaccine trials.

Antigen polymorphisms in essential malarial antigens are a key challenge to the design and development of broadly effective malaria vaccines. The effect of polymorphisms on antibody responses is fairly well studied while much fewer studies have assessed this for T cell responses. This study investigated the effect of allelic polymorphisms in the malarial antigen apical membrane antigen 1 (AMA1) onex vivoT cell-specific IFN-γ responses in subjects with lifelong exposure to malaria. Human leukocyte antigen (HLA) class I-restricted peptides from the 3D7 clone AMA1 were bioinformatically predicted and those with variant amino acid positions used to select corresponding allelic sequences from the 7G8, FVO, FC27 and tm284 parasite strains. A total of 91 AMA1 9-10mer peptides from the five parasite strains were identified, synthesized, grouped into 42 allele sets and used to stimulate PBMCs from seven HLA class 1-typed subjects in IFN-γ ELISpot assays. PBMCs from four of the seven subjects (57%) made positive responses to 18 peptides within 12 allele sets. Fifty percent of the 18 positive peptides were from the 3D7 parasite variant. Amino acid substitutions that were associated with IFN-γ response abrogation were more frequently found at positions 1 and 6 of the tested peptides, but substitutions did not show a clear pattern of association with response abrogation. Thus, while we show some evidence of polymorphisms affecting T cell response induction, other factors including TCR recognition of HLA-peptide complexes may also be at play.

Background: National immunization programs around the world have seen massive improvements in the last few decades especially, in vaccinations coverage. This notwithstanding, vaccine-preventable diseases (VPDs) have persisted globally with a disproportionately high burden in Low and Middle-Income Countries (LMICs). Perhaps, a more nuanced understanding of VPD transmission beyond vaccination coverage may help optimise VPDs control efforts. This study set out to first identify VPDs hotspots in Ghana, and subsequently, explore how ecology, ubanicity and land-use variations (Artisanal and Small-scale Gold Mining (ASGM) activities) may influence the spatial distribution hotspots. Methods: The study used district-level disease count data from 2010 to 2014 from the Ghana Health Service (GHS) and population data from the Ghana Population and Housing Census (PHC) in determining the clustering patterns for six VPDS (Measles, Meningitis, Mumps, Otitis media, Pneumonia and Tetanus). Purely spatial and space-time cluster analyses were implemented in SaTScan using the discrete Poisson model and the default settings which calculate pp = 0.045), incidence of the other five VPDs did not differ across the urban-rural divide. The relationship between ASGM activities and VPDs was ambivalent. Whereas spatial analysis suggested that some VPD hotspots (tetanus and otitis media) occur more frequently in mining districts in the southern part of the country, a Mann-Whitney U test revealed a higher incidence of meningitis in non-mining districts (p = 0.019). Interpretation: Taken together, this study presents evidence of a preponderance of VPDs hotspots in the northern parts of Ghana. It also highlights differences in VPD clustering patterns and its implications for integration VPDs control activities into a unified framework. Further studies will be needed to clarify the relationship between ASGM activities and VPD transmission in Ghana. Funding Information: This study was partly supported by funding from Pure Earth. Declaration of Interests: We declare no competing interests.

Findings from previous comparative genomics studies of the Mycobacterium tuberculosis complex (MTBC) suggest genomic variation among the genotypes may have phenotypic implications. We investigated the diversity in the phenotypic profiles of the main prevalent MTBC genotypes in West Africa. Thirty-six whole genome sequenced drug susceptible MTBC isolates belonging to lineages 4, 5 and 6 were included in this study. The isolates were phenotypically characterized for urease activity, tween hydrolysis, Thiophen-2-Carboxylic Acid Hydrazide (TCH) susceptibility, nitric oxide production, and growth rate in both liquid (7H9) and solid media (7H11 and Löwenstein–Jensen (L-J)). Lineage 4 isolates showed the highest growth rate in both liquid (p = 0.0003) and on solid (L-J) media supplemented with glycerol (p

Nearly 60% of Plasmodium falciparum proteins are still uncharacterized and their functions are unknown. In this report, we carried out the functional characterization of a 45 kDa protein (PF3D7_1459400) and showed its potential as a target for blood stage malaria vaccine development. Analysis of protein subcellular localization, native protein expression profile, and erythrocyte invasion inhibition of both clinical and laboratory parasite strains by peptide antibodies suggest a functional role of PF3D7_1459400 protein during erythrocyte invasion. Also, immunoreactivity screens using synthetic peptides of the protein showed that adults resident in malaria endemic regions in Ghana have naturally acquired plasma antibodies against PF3D7_1459400 protein. Altogether, this study presents PF3D7_1459400 protein as a potential target for the development of peptide-based vaccine for blood-stage malaria. IMPACT STATEMENT: Plasmodium falciparum malaria is a global health problem. Erythrocyte invasion by P. falciparum merozoites appears to be a promising target to curb malaria. We have identified and characterized a novel protein that is involved in erythrocyte invasion. Our data on protein subcellular localization, stage-specific protein expression pattern, and merozoite invasion inhibition by α-peptide antibodies suggest a role for PF3D7_1459400 protein during P. falciparum erythrocyte invasion. Even more, the human immunoepidemiology data present PF3D7_1459400 protein as an immunogenic antigen which could be further exploited for the development of new anti-infective therapy against malaria.

The continuous spread of antimalarial drug resistance is a threat to current chemotherapy efficacy. Therefore, characterizing the genetic diversity of drug resistance markers is needed to follow treatment effectiveness and further update control strategies. Here, we genotyped Plasmodium falciparum resistance gene markers associated with sulfadoxine-pyrimethamine (SP) and artemisinin-based combination therapy (ACT) in isolates from pregnant women in Ghana. The prevalence of the septuple IRN I- A/FG K GS/Tpfdhfr/pfdhps haplotypes, including the pfdhps A581G and A613S/T mutations, was high at delivery among post-SP treatment isolates (18.2%) compared to those of first antenatal care (before initiation of intermittent preventive treatment of malaria in pregnancy with sulfadoxine-pyrimethamine [IPTp-SP]; 6.1%; P = 0.03). Regarding the pfk13 marker gene, two nonsynonymous mutations (N458D and A481C) were detected at positions previously related to artemisinin resistance in isolates from Southeast Asia. These mutations were predicted in silico to alter the stability of the pfk13 propeller-encoding domain. Overall, these findings highlight the need for intensified monitoring and surveillance of additional mutations associated with increased SP resistance as well as emergence of resistance against artemisinin derivatives.

Preeclampsia (PE) is a placental disorder with different phenotypic presentations. In malaria-endemic regions, high incidence of PE is reported, with debilitating foeto-maternal effects, particularly among primigravid women. However, the relationship between placental pathology and Plasmodium falciparum infection in the placenta with PE is underexplored. Placentas from 134 pregnant women were examined after delivery for pathological lesions and placental malaria (PM). They comprised of 69 women without PE (non-PE group) and 65 women diagnosed with PE (PE group). The presence of placental pathology increased the risk of PE, with particular reference to syncytial knots. Placental malaria was 64 (48.1%) and 21 (15.8%) respectively for active and past infections and these proportions were significantly higher in the PE group compared to the non-PE group. Further multivariate analyses showed placental pathology (adjusted (aOR) 3.0, 95% CI = 1.2–7.5), active PM (aOR 6.7, 95% CI = 2.3–19.1), past PM (aOR 12.4, 95% CI = 3.0–51.0) and primigravidity (aOR 6.6, 95% CI 2.4–18.2) to be associated with PE. Our findings suggest that placental histological changes and PM are independent risk factors for PE particularly in primigravida. These findings might improve the management of PE in malaria-endemic regions.

Chronic hepatitis B infection is an important medical problem in sub-Saharan Africa. With increasing concerns of dwindling access to needed care, increasing cost of treatment, and rising prevalence of dire outcomes like liver cirrhosis and hepatocellular cancer, the need to determine the genetic associations underpinning hepatitis B virus persistence or clearance in a population comes to the fore. Genetic association studies have suggested a variation in human leukocyte antigen alleles associated with hepatitis B virus outcome along geo-ethnic lines. We investigated the association of human leukocyte antigen alleles to hepatitis B virus outcome against this backdrop. We used targeted next generation sequencing to type the human leukocyte antigen class I and II alleles of 173 study participants. These comprised of 92 cases with chronic hepatitis B infection and 81 healthy controls with serological evidence of naturally cleared hepatitis B virus infection. We have identified human leukocyte antigen alleles associated with hepatitis B virus clearance and persistence for the first time in a Ghanaian population. The class 1 allele C*16:01 (odds ratio (OR) = 3.4, confidence interval (CI) = 1.6–7.0, P-value = 0.01) was associated with hepatitis B virus persistence. Four class I alleles and one class II allele: A*34:02 (OR = 0.1, CI = 0.04–0.2, P-value = 3.4e-05), A*74:01 (OR = 0.3, CI = 0.2–0.7, P-value = 0.0135), B*13:02 (OR = 0.04, CI = 0.01–0.2, P-value = 0.000172), C*08:04 (OR = 0.06, CI = 0.01–0.2, P-value = 7.83e-05), and DRB1*08:04 (OR = 0.2, CI = 0.03–0.27, P-value = 0.000252) were associated with hepatitis B virus clearance. Our data show that previously reported human leukocyte antigen alleles associations to hepatitis B virus outcome are not found in this Ghanaian study. This study has therefore identified human leukocyte antigen types that are associated with either hepatitis B virus persistence or clearance and highlights the importance of geo-ethnic pivoted studies in determining the genetic associations to acute hepatitis B virus infection outcome. Impact statement Genetic association studies can determine the effect size of gene loci on disease outcomes. In the arena of HBV infections, HLA alleles that associate with HBV outcomes can be used in clinical management decisions. This potential translational utility can shape the future management of HBV infections by identifying at-risk individuals and tailoring medical interventions accordingly. This precision medicine motif is currently only a nascent idea. However, it has stakes that may well override the current “wait and see” approach of clinical management of HBV infections. Here, we have identified HLA alleles associated with HBV outcome in a Ghanaian cohort. Our findings support the motif that HLA alleles associate with HBV outcome along geo-ethnic lines. This buttresses the need for further population pivoted studies. In the long term, our findings add to efforts towards the development of an HLA molecular-based algorithm for predicting HBV infection outcomes.