Metrics details
Recent reports suggest that cell-surface and intracellular immune receptors function synergistically to activate robust defence against pathogens
Here we determined the numbers of cell-surface and intracellular immune receptors in 350 species
the number of receptor genes that are predicted to encode cell-surface and intracellular immune receptors is strongly correlated
We suggest this is consistent with mutual potentiation of immunity initiated by cell-surface and intracellular receptors being reflected in the concerted co-evolution of the size of their repertoires across plant species
PTI and ETI function synergistically to provide robust immunity against pathogens
As PRRs and NLRs are functionally inter-dependent
we investigated whether the sizes of these two receptor gene families are correlated
Phylogenetic tree of 350 plant species
79 monocot species and 208 eudicot species
Heat maps represent the percentages (%) of LRR-RLKs
LRR-RLK-XIIs + LRR-RLPs (magenta) and NB-ARCs (blue) in their corresponding annotated proteomes
Grey boxes in heat maps indicate null values where no receptors were identified
Scatter plot of %LRR-RLP + LRR-RLK-XII against %NB-ARC
with dark-grey shade representing the 95% confidence interval and light-grey shade representing the 95% prediction interval
aquatic species and trees are indicated as yellow inverted triangles
Model organisms are also indicated as spheres of different colours
Schematic illustration of the co-expansion and co-contraction of immune receptors in plant genomes
we conclude that PRR gene families specifically involved in pathogen recognition co-expand or co-contract with NB-ARC gene families
thomaeum has a similar number of predicted proteins as other Poales species such as Ananas comosus
suggesting that the contraction of immune receptor families could be independent of the reduced genome size
some plant lifestyles might also correlate with expansion of immune receptor gene families
As %LRR-RLK-III and %LRR-RLK-XI do not show positive correlation with %NB-ARC
while NB-ARC-encoding genes can form genomic clusters adjacent to LRR-RLK-XIIs
the co-expansion/contraction of these immune receptors is likely to be caused by mechanisms other than genomic clustering
partial or complete submersion of aquatic species results in reduced exposure to airborne pathogenic spores
removing an interface for interaction with pathogens
Lifespan may also drive changes in the immune receptor repertoire
We found that trees generally show higher %PRR and %NB-ARC than other species
While annual plants are subject to shorter periods of pathogen pressure before reproduction
this long-term pathogen pressure could drive the expansion of immune receptor gene families
inbreeding species may require an increased number of immune receptors compared with their outbreeding ancestors
an outcome that can also result from polyploidy
As the concerted expansion and contraction of immune receptors in plant genomes is not due to genomic clustering
further study is needed to understand the mechanism(s) underpinning these observations
As functionally inter-dependent genes often co-expand/contract together
it is likely that the functional relationship between cell-surface and intracellular immune receptors is conserved across plant species
NB-ARCs were identified using the set of proteins with a minimal length of 150 AA
Proteins were then searched for the presence of NB-ARC (PF00931.23) domains (hmmer option -E 1e-10 for NB-ARC)
Candidates were split into LysM-RLKs and LysM-RLPs by searching for presence/absence of a kinase domain (PF00069.26
The latter contained 238 out of the 350 genomes analysed
Statistical analyses were performed with OriginPro (version 2022; https://www.originlab.com/) and R (version 3.4.4)
Further information on research design is available in the Nature Research Reporting Summary linked to this article
All the analyses were done as described in Methods with publicly available tools (hmmer, tmhmm, diamond, FastTree, FAMSA, gotree and R). Scripts are available on github.com/MWSchmid/Ngou-et-al.-2022
Thirty years of resistance: zig-zag through the plant immune system
Mutual potentiation of plant immunity by cell-surface and intracellular receptors
Pattern-recognition receptors are required for NLR-mediated plant immunity
The EDS1-PAD4-ADR1 node mediates Arabidopsis pattern-triggered immunity
Activation of TIR signalling boosts pattern-triggered immunity
and exploitation of plant pattern recognition receptors for broad-spectrum disease resistance
Expansion of the receptor-like kinase/Pelle gene family and receptor-like proteins in Arabidopsis
Origin and diversity of plant receptor-like kinases
Evolutionary history and stress regulation of plant receptor-like kinase/pelle genes
Arabidopsis transmembrane receptor-like kinases (RLKs): a bridge between extracellular signal and intracellular regulatory machinery
The sequenced genomes of nonflowering land plants reveal the innovative evolutionary history of peptide signaling
Intracellular innate immune surveillance devices in plants and animals
Long-term evolution of nucleotide-binding site-leucine-rich repeat genes: understanding gained from and beyond the legume family
Convergent loss of an EDS1/PAD4 signaling pathway in several plant lineages reveals coevolved components of plant immunity and drought response
New insights on leucine-rich repeats receptor-like kinase orthologous relationships in angiosperms
RGAugury: a pipeline for genome-wide prediction of resistance gene analogs (RGAs) in plants
Quinone perception in plants via leucine-rich-repeat receptor-like kinases
Hydrogen peroxide sensor HPCA1 is an LRR receptor kinase in Arabidopsis
is required for cellooligomer-induced responses in Arabidopsis thaliana
Loss-of-function of Arabidopsis receptor-like kinase BIR1 activates cell death and defense responses mediated by BAK1 and SOBIR1
a tool for generating plant phylogenies and an analysis of phylogenetic community structure
An angiosperm NLR Atlas reveals that NLR gene reduction is associated with ecological specialization and signal transduction component deletion
Single-molecule sequencing of the desiccation-tolerant grass Oropetium thomaeum
Overview of tomato (Solanum lycopersicum) candidate pathogen recognition genes reveals important Solanum R locus dynamics
Resistance gene analogs in the Brassicaceae: identification
Oak genome reveals facets of long lifespan
The role of hybridization in the evolution and emergence of new fungal plant pathogens
Emergence of the Ug99 lineage of the wheat stem rust pathogen through somatic hybridisation
Sexual reproduction as an adaptation to resist parasites (a review)
Fast and sensitive protein alignment using DIAMOND
FAMSA: fast and accurate multiple sequence alignment of huge protein families
Current methods for automated filtering of multiple sequence alignments frequently worsen single-gene phylogenetic inference
FastTree 2—approximately maximum-likelihood trees for large alignments
Gotree/Goalign: toolkit and Go API to facilitate the development of phylogenetic workflows
Multi-omics approach highlights differences between RLP classes in Arabidopsis thaliana
Phylogenomic analysis of the receptor-like proteins of rice and Arabidopsis
Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes
Interactive Tree Of Life (iTOL) v5: an online tool for phylogenetic tree display and annotation
Controlling the false discovery rate: a practical and powerful approach to multiple testing
Ngou, B. P. M., Heal, R., Wyler, M., Schmid, M. W. & Jones, J. D. Genome-wide identification of cell-surface and intracellular immune receptors in 350 plant species. Zenodo https://doi.org/10.5281/zenodo.7017981 (2022)
Download references
was supported by the Norwich Research Park Biosciences Doctoral Training Partnership from the Biotechnology and Biological Sciences Research Council (BBSRC) (grant agreement BB/M011216/1)
These authors contributed equally: Bruno Pok Man Ngou
RIKEN Center for Sustainable Resource Science
conceived and conceptualized the study; B.P.M.N.
designed and performed the bioinformatic analyses; B.P.M.N.
performed the statistical analyses; B.P.M.N
The authors declare no competing interests
reviewer(s) for their contribution to the peer review of this work
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
ETI is activated by estradiol-induced expression of AvrRps4 in Arabidopsis thaliana for 4 hours
Number of genes (n) from each LRR-RLK subgroup: I
with heatmaps representing the assembled genome size
number (no.) of annotated proteins and number (no.) of primary transcripts
Brown branches indicates monocots and teal branches represent eudicots
Protein sequences from all 350 proteomes were first filtered for the primary gene models
Primary-transcript proteomes were then filtered and NB-ARC
LysM-RLP and LRR-RLK proteins were identified
LRR-RLK genes were further classified into 20 subgroups according to their alignment to the Arabidopsis thaliana subgroups
Details of the pipeline are described in the methods section
the black line represents the linear trend
with dark grey shade represents the 95% confidence interval and light grey shade represents the 95% prediction interval
Bottom left boxes include scatter plot between the corresponding % receptor-gene families in 300 angiosperms
Grey dots represent basal angiosperms (n = 13)
brown dots represent monocots (n = 79) and green dots represent eudiots (n = 208)
The Pearson correlation coefficient (Pearson’s r) is indicated below each scatter plot
The diagonal boxes include the distribution of % receptor-gene families in 300 angiosperms
Phylogenetic tree of the species used in the analysis
Pearson correlation between %LRR-RLK_XII+LRR-RLP and %NB-ARC in b
Phylogenetic tree of the Asterids clade used in the analysis
Carnivorous plants are marked with orange stars; parasitic plants are marked with yellow triangles and aquatic plants are marked with blue circles
Pearson correlation between %LRR-RLK_XII+LRR-RLP and %NB-ARC in the Asterids clade
Number of species (n) in each category: non-parasitic species
Table summarizing the statistical analysis of genomic clustering between PRRs (LRR-RLKs and LRR-RLPs) and NB-ARCs in Solanum tuberosum (a)
The 90-percentile distance (bp) between PRR gene family members and the next closest NB-ARC genes were calculated
This is then compared to a distribution (n = 1000) of 90-percentile distances between randomly-sampled genes and the next closest NB-ARC genes
One-sided test was performed to test the differences between tested distance (PRRs) and sampled distance (randomly-sampled)
P-values are calculated based on the comparison to 1000 cases of randomly-sampled genes
Significant values are indicated in bold (p-value < 0.05 is considered as significant)
Distribution (n = 1000) of 90-percentile distances (bp) between randomly-sampled genes and the next closest NB-ARC genes in Solanum tuberosum (b)
Red lines indicate the 90-percentile distance between the corresponding PRR gene family members and the next closest NB-ARC genes
Percentage of receptor genes in each species
Download citation
DOI: https://doi.org/10.1038/s41477-022-01260-5
Anyone you share the following link with will be able to read this content:
a shareable link is not currently available for this article
Sign up for the Nature Briefing newsletter — what matters in science
Metrics details
but that mediated by intracellular receptors has rarely been investigated in the absence of surface-receptor-mediated immunity
interactions between these two immune pathways are poorly understood
by activating intracellular receptors without inducing surface-receptor-mediated immunity
we analyse interactions between these two distinct immune systems in Arabidopsis
Pathogen recognition by surface receptors activates multiple protein kinases and NADPH oxidases
and we find that intracellular receptors primarily potentiate the activation of these proteins by increasing their abundance through several mechanisms
the hypersensitive response that depends on intracellular receptors is strongly enhanced by the activation of surface receptors
Activation of either immune system alone is insufficient to provide effective resistance against the bacterial pathogen Pseudomonas syringae
immune pathways activated by cell-surface and intracellular receptors in plants mutually potentiate to activate strong defences against pathogens
These findings reshape our understanding of plant immunity and have broad implications for crop improvement
Prices may be subject to local taxes which are calculated during checkout
Regulation of pattern recognition receptor signalling in plants
and genomic diversity of plant NLR proteins: an evolved resource for rational engineering of plant immunity
Reconstitution and structure of a plant NLR resistosome conferring immunity
Structure of the activated ROQ1 resistosome directly recognizing the pathogen effector XopQ
Direct pathogen-induced assembly of an NLR immune receptor complex to form a holoenzyme
A comparative overview of the intracellular guardians of plants and animals: NLRs in innate immunity and beyond
A coevolved EDS1-SAG101-NRG1 module mediates cell death signaling by TIR-domain immune receptors
Estradiol-inducible AvrRps4 expression reveals distinct properties of TIR-NLR-mediated effector-triggered immunity
Callose-mediated resistance to pathogenic intruders in plant defense-related papillae
Direct regulation of the NADPH oxidase RBOHD by the PRR-associated kinase BIK1 during plant immunity
The FLS2-associated kinase BIK1 directly phosphorylates the NADPH oxidase RbohD to control plant immunity
MAPK cascades in plant disease resistance signaling
Defended to the nines: 25 years of resistance gene cloning identifies nine mechanisms for R protein function
Translatome analysis of an NB-LRR immune response identifies important contributors to plant immunity in Arabidopsis
High-resolution expression profiling of selected gene sets during plant immune activation
Ding, P. et al. Chromatin accessibility landscapes activated by cell surface and intracellular immune receptors. Preprint at https://doi.org/10.1101/2020.06.17.157040 (2020)
Ligand-induced monoubiquitination of BIK1 regulates plant immunity
Regulation of reactive oxygen species during plant immunity through phosphorylation and ubiquitination of RBOHD
Translational gene regulation in plants: a green new deal
The Arabidopsis leucine-rich repeat receptor-like kinases BAK1/SERK3 and BKK1/SERK4 are required for innate immunity to hemibiotrophic and biotrophic pathogens
Yuan, M. et al. Pattern-recognition receptors are required for NLR-mediated plant immunity. Nature, https://doi.org/10.1038/s41586-021-03316-6 (2021)
Active photosynthetic inhibition mediated by MPK3/MPK6 is critical to effector-triggered immunity
Arabidopsis gp91phox homologues AtrbohD and AtrbohF are required for accumulation of reactive oxygen intermediates in the plant defense response
Quantitative phosphoproteomic analysis reveals common regulatory mechanisms between effector- and PAMP-triggered immunity in plants
A chemical genetic approach demonstrates that MPK3/MPK6 activation and NADPH oxidase-mediated oxidative burst are two independent signaling events in plant immunity
Regulation of sugar transporter activity for antibacterial defense in Arabidopsis
Decreased abundance of type III secretion system-inducing signals in Arabidopsis mkp1 enhances resistance against Pseudomonas syringae
Putting knowledge of plant disease resistance genes to work
Luo, M. et al. A five-transgene cassette confers broad-spectrum resistance to a fungal rust pathogen in wheat. Nat. Biotechnol. https://doi.org/10.1038/s41587-020-00770-x (2021)
Transgressive segregation reveals mechanisms of Arabidopsis immunity to Brassica-infecting races of white rust (Albugo candida)
The NLRP3 inflammasome: an overview of mechanisms of activation and regulation
Peptidoglycan recognition by the innate immune system
Phosphorylation-dependent differential regulation of plant growth
and innate immunity by the regulatory receptor-like kinase BAK1
A downy mildew effector evades recognition by polymorphism of expression and subcellular localization
Large-scale structure-function analysis of the Arabidopsis RPM1 disease resistance protein
Dual regulation of gene expression mediated by extended MAPK activation and salicylic acid contributes to robust innate immunity in Arabidopsis thaliana
Recognition of the protein kinase AVRPPHB SUSCEPTIBLE1 by the disease resistance protein RESISTANCE TO PSEUDOMONAS SYRINGAE5 is dependent on S-acylation and an exposed loop in AVRPPHB SUSCEPTIBLE1
Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method
Near-optimal probabilistic RNA-seq quantification
Guo, W. et al. 3D RNA-seq - a powerful and flexible tool for rapid and accurate differential expression and alternative splicing analysis of RNA-seq data for biologists. RNA Biol. https://doi.org/10.1080/15476286.2020.1858253 (2020)
Pescadillo plays an essential role in plant cell growth and survival by modulating ribosome biogenesis
Proteomic analysis of SUMO1-SUMOylome changes during defense elicitation in Arabidopsis
requires in planta processing and the KRVY domain to function
Recombineering and stable integration of the Pseudomonas syringae pv
syringae 61 hrp/hrc cluster into the genome of the soil bacterium Pseudomonas fluorescens Pf0-1
Download references
He for critical reading of the manuscript; and the Gatsby Foundation for funding to the J.D.G.J
B.P.M.N was supported by the Norwich Research Park Biosciences Doctoral Training Partnership from the Biotechnology and Biological Sciences Research Council (BBSRC) (grant agreement BB/M011216/1); H.-K.A
was supported by European Research Council Advanced Grant ‘ImmunitybyPairDesign’ (grant agreement: 669926); and P.D
acknowledges support from the European Union’s Horizon 2020 Research and Innovation Program under Marie Skłodowska-Curie Actions (grant agreement 656243) and a Future Leader Fellowship from BBSRC (grant agreement BB/R012172/1)
Present address: Institute of Biology Leiden
conceived and conceptualized the study; B.P.M.N
HR assay and electrolyte leakage assay; B.P.M.N performed the extraction of plasma membrane proteins with assistance from H.-K.A.; H.-K.A designed and performed the cycloheximide and MG132 experiment; H.-K.A
Peer review information Nature thanks Thorsten Nürnberger and the other
All experiments were repeated at least three times with similar results
Source data
Tabular summary of total ROS production in different phases after different PAMP or DAMP treatments with co-activation of PTI and ETIAvrRps4
Source data
CERK1 and MPK4 after activation of ETI for 4 h and 8 h in several effector-inducible lines
AvrRps4 and AvrRpp4 lines were infiltrated with 50 μM dexamethasone (for Dex:AvrRpm1) or 50 μM oestradiol
4 h and 8 h after infiltration for protein extraction
Molecular weight markers (in kDa) are indicated on the left
Ponceau staining (PS) was used as loading control
Source data
Source data
Source data
Ponceau staining was used as loading control
Source data
Source data
Source data
nlp20 or chitin does not lead to macroscopic HR
Co-activation of PTI (triggered by these PAMPs or DAMP) with ETIAvrRps4 leads to macroscopic HR
The numbers indicate the number of leaves that display HR of the total number of leaves infiltrated
Five-week-old inducible AvrRpm1 (Dex:AvrRpm1)
AvrRps4 (Est:AvrRps4) and AvrRpp4 (Est:AvrRpp4) Arabidopsis leaves were infiltrated with either dexamethasone (for Dex:AvrRpm1 only) or oestradiol
The combination of PTI and ETI leads to stronger macroscopic HR in inducible AvrRpm1
Source data
MPK phosphorylation during ETI triggered by multiple effectors
Est:AvrPphB and Est:AvrRpp4 lines were soaked in dexamethasone or oestradiol solution
Untreated seedlings were used as a negative control; seedlings treated with 100 nM flg22 for 15 min (red
RBOHD phosphorylation during ETI triggered by multiple effectors
Est:AvrPphB and Est:AvrRpp4 were soaked in mock (black)
dexamethasone or oestradiol solution (dark yellow) for 6 h
Microsomal fractions from seedlings were isolated for immunoblotting with RBOHD(pS39) antibody
Ponceau staining was used as the loading control
MPK6SR#58 (mpk3 mpk6 PMPK6:MPK6YG) is a conditional mpk3 mpk6 double mutant
MPK6YG has a larger ATP-binding pocket than wild-type MPK6 and is sensitive to the inhibitor 1-naphthyl-PP1 (NA-PP1
Pre-treatment with NA-PP1 inhibits MPK6YG and temporarily generates a mpk3 mpk6 double mutant
Both Col-0 and MPK6SR#58 leaves were pre-infiltrated with either 1% DMSO (mock) or 10 μM NA-PP1
these leaves were infiltrated with either Pf0-1:empty vector (triggers PTI) or Pf0-1:AvrRps4 (triggers PTI + ETIAvrRps4)
Pf0-1:AvrRps4 infiltration leads to macroscopic HR in both Col-0 and MPKS6R#58
NA-PP1 pre-treatment attenuates HR caused by Pf0-1:AvrRps4 only in the MPK6SR#58 line
Col-0 and rbohd rbohf leaves were infiltrated with either Pf0-1:empty vector (triggers PTI) or Pf0-1:AvrRps4 (triggers PTI + ETIAvrRps4) at a varying OD600
Pf0-1:AvrRps4 infiltration leads to less macroscopic HR in rbohd rbohf leaves
Activation of an NLR (ETI without PTI) increases the accumulation of PTI signalling components
Co-activation of both PTI and ETI increases this accumulation and enhances the activation of multiple PTI signalling components
Molecular weight in kDa (kilodalton) is indicated on the left
This file contains Supplementary Tables 1-3
Differentially expressed gene list during ETI
List of differentially expressed (DE) genes identified in the comparison between Est:AvrRps4 treated with estradiol for 0h (seti_e2_0h) and 4h (seti_e2_4h)
Positive log2FC values (red) indicates upregulation and negative log2FC values (blue) indicates downregulation
P-values for differentially expressed (DE) genes were generated with Fisher Z-transformation after Student’s t-test
DE genes with “Benjamini and Hochberg’s (BH) method” false discovery rate (FDR) two-sided adjusted P-value (adj.pval) < 0.05 (green) is categorized as significant
Summary of all statistical analysis are listed
The specification of samples and the number of samples used for each dataset
and the statistical tests and the P-values are summarized
The methods used for adjustment of P-values are also shown
The source data of all the statistical analysis can be found in the Source File
Download citation
DOI: https://doi.org/10.1038/s41586-021-03315-7
Plants are continuously evolving new immune receptors to ever-changing pathogens
Researchers at the RIKEN Center for Sustainable Resource Science (CSRS) have traced the origin and evolutionary trajectory of plant immune receptors
Their discovery will make it easier to identify immune receptor genes from genomic information and could help in the development of pathogen-resistant crops
This study was published in the scientific journal Nature Communications on February 1
plants have immune responses that help them defend against pathogens such as viruses
and this is accomplished by pattern recognition receptors located on the surface of plant cells
The ability of these receptors to detect molecular patterns associated with pathogens depends on two types of proteins
both of which can contain leucine-rich repeats—sections in which the amino acid leucine appears multiple times
the international research team led by Ken Shirasu and Yasuhiro Kadota at RIKEN CSRS examined the numbers and patterns of receptors
They analyzed over 170,000 genes encoding RLKs and about 40,000 genes encoding RLPs
which they obtained from publicly available data taken from 350 plant species
They discovered that RLKs and RLPs with leucine-rich repeats were the most abundant receptor types among all the plant species
making up nearly half of RLKs and 70% of RLPs
are known to contain a special island region that is crucial for recognizing parts of pathogens
Investigation by the RIKEN CSRS team revealed that amongRLPsthat contain the leucine-rich repeats
this special region was almost always located in the same place; between the 4th and 5th leucine-rich repeat
These RLPs were found to be associated with immune responses
They also discovered that the island region was located at the same position in some RLKs
nearly all of which belong to a functional group that regulates growth and development
Comparative analysis showed that the sequence of the four repeats below the island region was very similar between the two types of protein detectors
suggesting that they have a common evolutionary ancestry
these four sets of leucine repeats contained sections needed for bonding to the same co-receptor
This means that immunity-related RLPs and growth-related RLKs inherited the ability to bind BAK1 from a common ancestor
we found that exchanging the four regions of leucine-rich repeats among these receptors did not disrupt their functionality,” says Bruno Pok Man Ngou
Creating a hybrid receptor by combining a growth-related RLK with an immunity-related RLP resulted in a hybrid receptor that recognized pathogens and induced both immune and growth-related responses
This means that scientists should be able to engineer receptors with new functions by swapping those modules
This study addressed the origins of plant immunity at a molecular level
showing that simultaneously analyzing information from multiple plant genomes can allow straightforward and precise prediction of genes involved in plant immunity and growth
“We are currently isolating immune receptors from various plants using this information
aiming for practical applications such as developing disease-resistant crops in the future,” says Shirasu
Masataka Sasabe RIKEN International Affairs Division Tel: +81-(0)48-462-1225 Email: masataka.sasabe [at] riken.jp
Both immunity-related LRR-RLPs and growth-related LRR-RLKs have evolved from a common ancestor to inherit the last four LRRs and the ability to bind the co-receptor BAK1
The chimeric receptor that incorporates the cytoplasmic kinase domain of a growth-related LRR-RLK into an immunity-related LRR-RLP activates both immune and growth responses upon sensing a pathogen-derived molecule
Top
Forbes contributors publish independent expert analyses and insights
Christine Ro is a journalist covering science and development
ShareSaveCommentA nurse prepares a dose of vaccine against malaria at a district hospital in Soa
More (Photo by Kepseu/Xinhua via Getty Images)
Prossy Muyingo is worried. Uganda announced an Ebola outbreak on Jan
and they help to plug the gap where nurses and doctors are in short supply
president and secretary of state abruptly cut off most aid
While people had been preparing for some sort of foreign aid review by the new administration
the rug-pulled-out part was that aid was immediately stopped while the review was pending
The State Department’s new spokesperson said that the 90-day cutoff of foreign aid was necessary because
they didn’t trust aid workers to provide information otherwise — even though continuation of funding typically depends on regular reporting
such as antiretroviral drugs to keep HIV in check
The aid freeze means that they will likely have to pay out of pocket — as well as going without the reminders and health information that are a key part of a CHW’s work
What will happen to them?” Muyingo wonders
She estimates that perhaps 2% of her patients will be able to afford the medicines they need
And she knows just how expensive these medicines are
as one of her children lives with sickle cell disease
Community health workers are already feeling the effects of the uncertainty, service cutoffs and loss of work, says Madeleine Ballard, the CEO of the Community Health Impact Coalition
both witnessing suffering and themselves experiencing suffering,” Ballard says
“I think it’s always the most devastating when those who care for others are not themselves cared for.”
there’s no clear line between emergency aid (which is allowed yet still ambiguous) and longer-term support that also saves lives
For instance, Olivia Ngou is the executive director of Impact Santé Afrique
which works from Cameroon to increase local political advocacy and community engagement around malaria
She hasn’t heard that malaria aid has been included in the lifesaving assistance exception
Delays are critical to ongoing malaria treatment and testing conducted by community health workers in remote areas
But it also affects months-long preparation for the rainy season
“The preparation for that has stopped,” Ngou reports
we are going to lose a lot of children.” Even more urgent is the 48-hour window to get tested and treated following a mosquito bite
The system for requesting waivers to the near-blanket freeze has also been confusing
Some organizations report a lack of clarity about where to even submit the requests
(The State Department did not immediately respond to a request for comment.)
the idea that contraception is unacceptable aid makes no sense to people who have seen loved ones die of AIDS
Muyingo has also seen the benefits of neutral
science-backed information on contraception: “Family planning has helped many students to finish their studies,” among other things
Some people don’t use condoms simply because they can’t afford them
the Heritage Foundation’s view that “education and abstinence could end the AIDS epidemic” has failed in the U.S
One phrase that’s being repeated over and over is “safer, stronger, and more prosperous.” This is the State Department’s mantra for justifiable aid. “Our test is simple,” the department’s spokesperson tweeted
Scientists have also overcome the challenges of developing vaccines for parasitic diseases
and vaccination against malaria is now routine in several African countries after starting in Cameroon
But vaccination programs have now halted due to the aid shutoff
sometimes before children have finished the full four-dose course of the malaria vaccine
Malaria No More estimates that a 90-day U.S
aid freeze could stop delivery of 15.6 million doses of medicine
trainings for 64,700 health workers and 9 million insecticide-treated mosquito nets
accounting for the contributions of all U.S
is that every day that the 90-day aid freeze continues
over 1,000 additional people will die of malaria
Much of U.S. aid on malaria comes through the U.S. President’s Initiative on Malaria, a nonpartisan program working in Africa and Southeast Asia. PMI has saved the lives of over 11 million people
Ngou says that it especially targets those at highest risk and in highest need: pregnant women and children under the age of 5
African countries have been stepping up to fight malaria, Ngou points out. In 2024, ministers of health from 11 African nations signed the Yaoundé Declaration
committing to putting more domestic resources to malaria control
And countries highly affected by malaria continue to build up health facilities and other infrastructure that U.S
While she and fellow advocates have been urging local leaders to release emergency funds to make up for aid gaps
“unfortunately the economic context right now is very challenging.”
there is talk of health and peace-building aid countering terrorism or Chinese influence
Some of the arguments have been pragmatic to the point of coldhearted
In an era when the State Department is proudly pointing to money being cut from aid
this detached approach may ultimately be effective
But bending over backward to justify certain aid projects as aligning with the new administration’s agenda risks undermining the core ethical imperative of aid
community health worker Muyingo agrees that her country will need more self-reliant health services
She believes that this will happen within the next decade; for now
the Ugandan government and other donors can’t fill the enormous gap left by the U.S
She would like the American government to remember that diseases have no borders
Metrics details
Cell-surface receptors play pivotal roles in many biological processes
How cell-surface receptors evolve to become specialised in different biological processes remains elusive
To shed light on the immune-specificity of cell-surface receptors
we analyzed more than 200,000 genes encoding cell-surface receptors from 350 genomes and traced the evolutionary origin of immune-specific leucine-rich repeat receptor-like proteins (LRR-RLPs) in plants
we discovered that the motifs crucial for co-receptor interaction in LRR-RLPs are closely related to those of the LRR-receptor-like kinase (RLK) subgroup Xb
which perceives phytohormones and primarily governs growth and development
Functional characterisation further reveals that LRR-RLPs initiate immune responses through their juxtamembrane and transmembrane regions
while LRR-RLK-Xb members regulate development through their cytosolic kinase domains
Our data suggest that the cell-surface receptors involved in immunity and development share a common origin
and cytosolic regions have either diversified or stabilised to recognise diverse ligands and activate differential downstream responses
Our work reveals a mechanism by which plants evolve to perceive diverse signals to activate the appropriate responses in a rapidly changing environment
the origins of PRR families involved in PAMP perception in plants remain largely unclear
Given the striking resemblance in their domain architecture
it is reasonable to infer that immunity- and development-related cell-surface receptors share a common origin
the evolutionary trajectory that led to their divergence and specialisation in distinct biological processes remains poorly understood
The bottom panel represents the presence or absence of different receptor classes in algal and plant lineages
‘-’ represents ectodomains with no transmembrane or kinase domain
‘RLP’ represents ectodomains with a transmembrane domain but no kinase domain
‘RLK’ represents ectodomains with both transmembrane and kinase domains
The number of species available from each algal and plant lineage is indicated by the numbers within respective boxes
A grey box indicates the absence of receptors and a green box indicates their presence in each lineage
The origin of a receptor is indicated with a circle (○)
Expansion rates of receptor classes are indicated by boxplots
The percentages (%) of cell-surface receptors from each genome were calculated as (number of identified genes/number of searched genes × 100)
the percentages from each species within a lineage (e.g.
Rhodophtya or green algae) were grouped and the median percentage was calculated
Median value was used instead of mean to avoid outliers within the lineages
The expansion rate within a species is calculated by ((% cell surface receptors in that species)-(median))/(median)
The cyan boxplot represents the expansion rate from Glaucophyta and Rhodophyta to green algae (LRR
The yellow boxplot represents the expansion rate from green algae to Embryophytes (LRR
n = 0) and the orange boxplot represents the differences between early land plants to Tracheophytes (LRR
Light blue area represents expansion and light pink area represents contraction of the gene family
X-axis values represent expansion rate (×)
Values larger than 0 indicate expansion; values equal to 0 indicate no expansion
reinforcing the idea that cell surface immune receptors underwent extensive expansions as the plant lineage diversified and evolved to adapt to a wide range of environments
The origin of a protein family is indicated with a circle (○)
followed by another circle indicating the origin of the orthologs of PTI-signalling component
Expansion rates of PTI-signalling component families are indicated by boxplots
The percentages (%) of signalling components from each genome were calculated as (number of identified genes/number of searched genes × 100)
the percentages from each species within a lineage (e.g
The expansion rate within a species is calculated by ((%signalling components in that species)-(median))/(median)
The cyan boxplot represents the expansion rate from Glaucophyta and Rhodophyta to green algae (SERKs
The yellow boxplot represents the expansion rate from green algae to Embryophytes (SERKs
n = 0) and the orange boxplot represents the differences between early land plants to Tracheophytes (SERKs
This observation implies a functional necessity for the specific placement of IDs in LRR-RLPs
d The concentric ring pie chart presents the percentage of LRR-containing cell-surface receptors (PRRs) from 350 species
The inner ring represents all LRR-containing cell-surface receptors (113,794); the middle ring represents LRR-containing PRRs with ID (20,556); the outer ring represents LRR-containing PRRs with an ID preceding the last 4 LRR (ID + 4LRR) at the C terminus (16,885)
e The presence or absence of receptor classes in various algal and plant lineages
and a green box indicates the presence of a given receptor class in each lineage
The origin of LRR-RLP and LRR-RLK-Xb with ID + 4LRR is indicated with a circle (○)
f Sequence similarity tree of the C3 region (last 4 LRRs) from all LRR-containing cell-surface receptors of 350 species
The inner ring and middle ring indicate the lineage and subclass/order of the corresponding protein (species) from the branch
Outer ring represents the LRR-RLP or LRR-RLK classification
The light grey area indicates clustering of LRR-RLK-Xb and LRR-RLP with ID + 4LRR
The pruned sequence similarity tree on the right g corresponds to the light grey area in the left tree
with clades labelled in dark grey areas accordingly
Characterized LRR-RLK-Xb and LRR-RLP members are labelled
The BRI/BRL-clade and the PSKR/PSY1R-clades are also labelled
Considering the similarity in structural motifs between LRR-RLPs and LRR-RLK-Xbs
it is likely that these two receptor classes share a common origin
e Alignment of amino acids in the last LRR motifs from NbRXEG1
Amino acid residues involved in the interaction between NbRXEG1 and BAK1 are highlighted in green
The QxxT motif positions are highlighted in yellow
Amino acids with similar properties to AtRLP23 are highlighted in grey
The last LRR motif of AtRLP23 is exchanged with either AtPSY1R
AtBRI1 have also been mutated to threonine (T)
l Immuno-precipitation to test interactions between AtRLP23 chimeras
Nb leaves expressing the indicated constructs were treated with either mock or 1 μM nlp20 for 5 min
h–k Functionality testing of AtRLP23 chimeras
Nb leaves expressing the indicated constructs were treated with 1 μM nlp20 and samples were collected at indicated time points
Phosphorylation of NbSIPK and NbWIPK was detected with p-P42/44 antibody
k Nb leaf discs expressing the indicated constructs were collected and treated with either mock or 1 μM nlp20
and ROS production was measured for indicated time points
the experiments were repeated at least twice with similar results
the residues around and within the TQxxx motif are both crucial for SERK interactions
We concluded that the C3 region in LRR-RLPs and some LRR-RLK-Xbs (such as PSY1R and PSKR1) interact with SERKs in a similar manner
while some LRR-RLK-Xbs (PSKR2 and BRI1) have evolved to interacts with SERKs in a slightly different manner
our results strongly support the functional conservation of C3 regions in LRR-RLK-Xbs and LRR-RLPs
specifically their ability to interact with SERKs
tracing back the origin of IDs is challenging due to their considerable diversity
We therefore propose that the IDs of LRR-RLK-Xbs and LRR-RLPs likely originated from a common ancestor
with the IDs of LRR-RLPs expanding and diversifying after the divergence of LRR-RLK-Xb and LRR-RLPs
We propose that these RLPs may either recognise endogenous molecules to activate growth and development
or participate in the recognition of pathogen-mimicking molecules to trigger immune signalling
Number of cell-surface receptors (n) in each LRR-RLK subgroup: I
Number of cell-surface receptors (n) in each LRR-RLP subgroup: LRR-RLP (RLP)
a The top panel depicts the presence of cell-surface receptors
and downstream signalling components (downstream) in Glaucophyta and Rhodophyta
Nodes are labelled in colours as indicated on the left
The absence of a node indicates the absence of a gene family from the lineage
Nodes with dotted outlines indicate the presence of a gene family
but the absence of immunity-related orthologs
Nodes with thick outlines indicate the expansion of gene families
Repeated expansion is indicated with thicker outlines
The middle panel (top) displays the percentages (%) of cell-surface receptors and signalling components in the genome of each species within the lineage
Middle panel (bottom) represents the distribution of signalling components
in the genome of each species within a lineage
Bars are labelled in colours as indicated on top left
The bottom panel shows examples of plant species and the classification of the plant lineages
Left panel: expansion of PRR family gene repertoires throughout the plant lineage
which leads to recognition of a larger range of PAMPs/MAMPs
and helper NLRs are absent from many algal species
A more complex immune network involving these signalling components apparently developed in vascular plants
Right panel: An ancient PRR with LRRID + 4LRR with unknown function evolved into LRR-RLK-Xbs and LRR-RLPs
which are involved in development- and immune-signalling
eJM-TM-cJM region of LRR-RLPs evolved to allow interactions with SOBIR1 to induce immunity (negatively charged eJM and GxxxG)
LRR-RLK-Xbs utilize Xb kinase domains to induce distinct downstream responses
The initial set of proteins included only the primary gene models from all 350 species (12,979,225 proteins in total)
sequences were filtered for a minimal length of 250 AA in the case of LRR-RLKs (7,690,505 proteins) or 150 AA in the case of all others (10,224,242 proteins)
Based on the presence of a kinase domain (KD) and/or a trans-membrane domain (TM)
the proteins fell into three major groups: (1) RLKs with KD and TM
and (3) ectodomain candidates without KD or TM
To identify RLPs and ectodomain candidates (without KD or TM)
we first removed all proteins with a kinase domain match (hmmer with the option -E1000)
leaving 9,746,585 from the initial 10,224,242 proteins
We next removed potential signalling peptides from the sequences with SignalP because they are sometimes identified as TM domains (trimmed 796,385 sequences)
We then searched for TMs with tmhmm and kept proteins with no TM (7,917,087
To test whether RLPs contained potentially functional endodomain sequences
we extracted endo- and ectodomain sequences
As tmhmm sometimes reverts in- and outside locations
we defined the stretch that was matched by the motif (e.g
the ectodomain sequence was defined as the stretch that contained the most LRR repeats
The endodomain was the remaining longest internal sequence stretch
The lengths of these sequences were visualized
We also searched for all available PFAM patterns in the endodomains of all RLPs as well as the proteins with kinase and TM domain with hmmsearch (with a strict e-value threshold of 1e-10 to ensure a minimum of partial matches)
we only kept the hit with the best score and counted the number of hits per PFAM domain and species
We only kept domains that were found in more than 1 or 1 out of 10,000 input sequences
Within the endodomains of the proteins with kinase and TM domains
99.42% were best matched by a kinase pattern
in all the RLPs we only found five kinase pattern matches among the 3'860 endodomains of WAK-RLPs (0.13%)
endodomains of RLPs were rarely matched by any PFAM pattern: 1.27% of the Duf26-RLPs
The WAK-RLPs were slightly different as 15.36% of their ectodomains matched to a PFAM pattern: most of them (13.94%) to a RING finger domain which might be involved in protein-protein interaction (PF13639.7)
and RBOHs were identified by hmmer searches (option -E 10) for specific domains
Ion transport protein domains (PFAM: PF00520.32) were used for CNGCs
PHM7_cyt (PFAM: PF14703.7) and RSN1_TM (PFAM: PF13967.7) were used for OSCAs
and FAD-binding domains (PFAM: PF08022.13)
ferric reductase like transmembrane components (PFAM: PF01794.20)
and ferric reductase NAD binding domains (PFAM: PF08030.13) were used for RBOHs
and Solyc02g067660) proteins were found in exactly one cluster each
we used the three matching clusters as EDS1
RPW8-NLRs (NRG1 and ADR1) were identified similarly using the NB-ARC (PFAM: PF00931.23) and RPW8 (PFAM: PF05659.12) domains
we found all known NRG1 and ADR1 sequences in one single cluster
This allowed us to extract and re-cluster these sequences with more stringent parameters (options --min-seq-id 0.3 -c 0.75)
and Niben101Scf02118g00018.1) and ADR1 (AT1G33560
and Niben101Scf02422g02015) proteins in exactly one cluster each
indicating that the two matching clusters could be considered as NRG1 and ADR1 proteins
The following patterns were used for the families of interest: SOBIR1: RLK-Pelle_LRR-XI-2
The percentages (%) of cell-surface receptors and signalling components from each genome were calculated as (number of identified genes/number of searched genes × 100)
The expansion rate within a species is calculated by ((% cell surface receptors or signalling components in that species)-(median))/(median)
the expansion rate of LRR-RLP family in Marchantia polymorpha from green algae is calculated by ((%LRR-RLP in Marchantia polymorpha)-(median %LRR-RLP in green algae)/(median %LRR-RLP in green algae)
Note that the reliability of the expansion rate is dependent on the number of species used to calculate the median
which is also dependent on the available genomes in Glaucophyta
We then pruned the sequences to include everything from the C3 domain to the C terminus
we further searched sequences for kinase domains and removed sequences upstream of the start of the kinase domain
C3 or eJM) were subsequently extracted from this alignment
the sequence similarity trees of specific domains were constructed as described above (FAMSA
In-depth analysis of the ectodomains from all Duf26
and WAK candidates was done using the ectodomain sequences extracted from the ectodomain-only proteins
Ectodomain sequences from the ectodomain-only proteins were extracted based on the location of the hmm-pattern match
Phylogenies were constructed as described above with FAMSA
and gotree (rooted with a sequence from the most basal species according to the NCBI taxonomy)
and AtBES1 were amplified by PCR with KoD one (Toyobo
and the PCR products were cloned into the epiGreenB5 (3× HA) vector between the ClaI and BamHI restriction sites with In-Fusion HD Cloning Kit (Clontech
USA) to generate p35S::BES1-HA or p35S::cell-surface receptor-HA (epiGreenB5-Cauliflower mosaic virus (CaMV) p35S:gene of interest-3 × HA)
The constructs were then transformed into Agrobacterium tumefaciens strain AGL1 for transient expression in Nicotiana benthamiana
All chimeric cell-surface receptors generated in this study contain the EFR signal peptide to ensure consistency between the constructs and expression levels
tumefaciens strain AGL1 carrying the binary expression vectors described above were grown on LB agar plates amended with selection antibiotics
and then resuspended in infiltration buffer (10 mM MgCl2
The concentration of AGL1 was then adjusted to OD600 = 0.5 and syringe-infiltrated into N
Protein extraction for immunoprecipitation was performed as previously described93
benthamiana leaves were treated with elicitors and snap-frozen
The tissues were then ground in liquid nitrogen and extracted in extraction buffer (50 mM Tris-HCl at pH 7.5
1% Protease Inhibitor Cocktail (P9599; Sigma-Aldrich)
100 μM phenylmethylsulphonyl fluoride and 2% IGEPAL CA-630 (v/v; Sigma-Aldrich)
and 2 mM EDTA) at a concentration of 3 mL/g tissue powder
Samples were then incubated at 4 °C for an hour and debris was removed by centrifugation at 13,000 rpm for 10 min at 4 °C
protein concentrations were adjusted to 5 mg/mL
then incubated with rotation for an hour at 4 °C with 50 μL anti-HA magnetic beads (Miltenyi Biotec) for immunoprecipitation
Magnetic beads were then washed twice with extraction buffer and the HA-tagged protein was eluted with sodium dodecyl sulphate (SDS) sample buffer at 95 °C
Protein extractions were performed as previously described94
benthamiana leaves were infiltrated with elicitors and snap-frozen at indicated time points
The tissues were then lysed in liquid nitrogen and extracted in 1×NuPAGE™ LDS Sample Buffer (Invitrogen™) with 10 mM DTT at 70 °C for 10 min
Total proteins were then separated by SDS-PAGE and blotted onto a nitrocellulose membrane (Trans-Blot Turbo Transfer System
The membrane was then blocked in a solution of either 5% skimmed milk (for BES1 and cell-surface receptor detection) or 5% bovine serum albumin (BSA; for MAPK detection) in Tris-buffered saline
0.1% Tween 20 detergent (TBST) for an hour
Phosphorylated MAPKs were detected using α-phospho-p44/42 MAPK rabbit monoclonal antibody (D13.14.4E
USA) in a solution of 5% BSA in TBST overnight at 4 °C
HA-tagged BES1 or cell surface receptors were detected using Anti-HA-Peroxidase
rat IgG1 antibody (Roche) in a solution of 5% skimmed milk in TBST overnight at 4 °C
this was followed by incubation with α-rabbit IgG-HRP-conjugated secondary antibodies (1:10,000
USA) in a solution of 5% BSA in TBST for an hour at room temperature
HRP signal was then detected by Clarity Western ECL Substrate (Bio-Rad) with a LAS 4000 system (GE Healthcare
Nitrocellulose membranes were stained with Coomassie Brilliant Blue (CBB) to ensure equal loading
ROS burst assays were performed as described previously93
benthamiana leaf discs were collected with a 4-mm-diameter cork borer and placed in 96-well plates with 120 μl deionised water overnight in the dark (abaxial surface of the leaves facing down)
benthamiana leaf discs were then treated with either mock (water) or 1 μM nlp20 in 20 mM luminol (Wako
Japan) and 0.02 mg ml−1 horseradish peroxidase (Sigma-Aldrich)
Luminescence was then measured over indicated periods of time with a Tristar2 multimode reader (Berthold Technologies
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article
Evolutionary dynamics of the leucine-rich repeat receptor-like kinase (LRR-RLK) subfamily in angiosperms
Concerted expansion and contraction of immune receptor gene repertoires in plant genomes
NLR surveillance of essential SEC-9 SNARE proteins induces programmed cell death upon allorecognition in filamentous fungi
Prokaryotic innate immunity through pattern recognition of conserved viral proteins
Innate immunity in plants and animals: emerging parallels between the recognition of general elicitors and pathogen-associated molecular patterns
Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases
Flourishing in water: the early evolution and diversification of plant receptor-like kinases
Phytocytokines function as immunological modulators of plant immunity
Plant cell surface receptor-mediated signaling - a common theme amid diversity
Out of water: the origin and early diversification of plant R-genes
Structural evolution drives diversification of the large LRR-RLK gene family
Molecular mechanisms of early plant pattern-triggered immune signaling
Roles of receptor-like cytoplasmic kinase VII members in pattern-triggered immune signaling
Signaling mechanisms in pattern-triggered immunity (PTI)
The structural basis of ligand perception and signal activation by receptor kinases
Receptor-like kinase SOBIR1/EVR interacts with receptor-like proteins in plant immunity against fungal infection
EDS1 signalling: at the nexus of intracellular and surface receptor immunity
Plant NLRs get by with a little help from their friends
Receptor-like cytoplasmic kinases directly link diverse pattern recognition receptors to the activation of mitogen-activated protein kinase cascades in Arabidopsis
Huang, W. R. H. et al. Receptor-like cytoplasmic kinases belonging to different subfamilies mediate immune responses downstream of the Cf-4 resistance protein in Nicotiana benthamiana. BioRxiv. https://doi.org/10.1101/2023.04.25.538242 (2023)
Tyrosine phosphorylation of the lectin receptor-like kinase LORE regulates plant immunity
The calcium-dependent protein kinase CPK28 buffers plant immunity and regulates BIK1 turnover
Calcium-dependent protein kinase/NADPH oxidase activation circuit is required for rapid defense signal propagation
Bifurcation of Arabidopsis NLR immune signaling via Ca2+-dependent protein kinases
Differential innate immune signalling via Ca(2+) sensor protein kinases
A Phytophthora capsici RXLR effector targets and inhibits the central immune kinases to suppress plant immunity
The Arabidopsis E3 ubiquitin ligase PUB4 regulates BIK1 and is targeted by a bacterial type-III effector
Receptor-like cytoplasmic kinases integrate signaling from multiple plant immune receptors and are targeted by a Pseudomonas syringae effector
Perturbations of the ZED1 pseudokinase activate plant immunity
Activation of a plant nucleotide binding-leucine rich repeat disease resistance protein by a modified self protein
The Arabidopsis ZED1 pseudokinase is required for ZAR1-mediated immunity induced by the Pseudomonas syringae type III effector HopZ1a
The decoy substrate of a pathogen effector and a pseudokinase specify pathogen-induced modified-self recognition and immunity in plants
Leucine-rich repeat (LRR) domains containing intervening motifs in plants
Plant Receptor-like proteins (RLPs): Structural features enabling versatile immune recognition
Arabidopsis leucine-rich repeat receptor-like kinase NILR1 is required for induction of innate immunity to parasitic nematodes
Signaling of cell fate determination by the TPD1 small protein and EMS1 receptor kinase
Tyrosine-sulfated glycopeptide involved in cellular proliferation and expansion in Arabidopsis
A LRR receptor kinase involved in perception of a peptide plant hormone
A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction
a receptor kinase pair mediating brassinosteroid signaling
Allosteric receptor activation by the plant peptide hormone phytosulfokine
An RLP23-SOBIR1-BAK1 complex mediates NLP-triggered immunity
Structure reveals that BAK1 as a co-receptor recognizes the BRI1-bound brassinolide
Structural basis of steroid hormone perception by the receptor kinase BRI1
Molecular mechanism for plant steroid receptor activation by somatic embryogenesis co-receptor kinases
Plant receptor-like protein activation by a microbial glycoside hydrolase
Structural insight into brassinosteroid perception by BRI1
Structural basis for flg22-induced activation of the Arabidopsis FLS2-BAK1 immune complex
Kinase activity of SOBIR1 and BAK1 is required for immune signalling
Peptide ligand-mediated trade-off between plant growth and stress response
Arabidopsis thaliana resistance to fusarium oxysporum 2 implicates tyrosine-sulfated peptide signaling in susceptibility and resistance to root infection
SOBIR1 requires the GxxxG dimerization motif in its transmembrane domain to form constitutive complexes with receptor-like proteins
BES1 accumulates in the nucleus in response to brassinosteroids to regulate gene expression and promote stem elongation
Evolutionary gain and loss of a plant pattern-recognition receptor for HAMP recognition
Pathogen-induced defense and innate immunity in macroalgae
The receptor kinase FER is a RALF-regulated scaffold controlling plant immune signaling
The leucine-rich repeat receptor kinase BIR2 is a negative regulator of BAK1 in plant immunity
The Arabidopsis leucine-rich repeat receptor kinase BIR3 negatively regulates BAK1 receptor complex formation and stabilizes BAK1
Structural basis for BIR1-mediated negative regulation of plant immunity
Regulation of cell death and innate immunity by two receptor-like kinases in Arabidopsis
Regulation of plant immune signaling by calcium-dependent protein kinases
Perception of brassinosteroids by the extracellular domain of the receptor kinase BRI1
Perception of a divergent family of phytocytokines by the Arabidopsis receptor kinase MIK2
The Arabidopsis MIK2 receptor elicits immunity by sensing a conserved signature from phytocytokines and microbes
Database mining of plant peptide homologues
A microbially derived tyrosine-sulfated peptide mimics a plant peptide hormone
Plant-parasitic nematode secreted peptides hijack a plant secretory pathway
SignalP 5.0 improves signal peptide predictions using deep neural networks
Identification and characterization of the LRR repeats in plant LRR-RLKs
Clustering huge protein sequence sets in linear time
classification and function of the plant protein kinase superfamily
LRRsearch: An asynchronous server-based application for the prediction of leucine-rich repeat motifs and an integrative database of NOD-like receptors
FAMSA: Fast and accurate multiple sequence alignment of huge protein families
ape 5.0: an environment for modern phylogenetics and evolutionary analyses in R
Fast unfolding of communities in large network
The igraph software package for complex network research | CiNii Research
Interactive Tree Of Life (iTOL) v5: an online tool for sequence similarity tree display and annotation
Mechanisms of RALF peptide perception by a heterotypic receptor complex
Chitin-induced dimerization activates a plant immune receptor
Highly accurate protein structure prediction with AlphaFold
a web-based 3D viewer for sharing 1D/2D/3D representations of biomolecular structures
Timetree 5: an expanded resource for species divergence times
Structural basis for differential recognition of brassinolide by its receptors
Download references
and Mizuki Yamamoto for providing technical support
and Jonathan Jones for discussions and critical reading of the manuscript
We also thank Markus Albert for sharing the information on chimeric receptors and BES1-HA experiments
is an International Research Fellow of the Japan Society for the Promotion of Science (Postdoctoral Fellowships for Research in Japan (Standard)
The research was financially supported by MEXT/JSPS KAKENHI Grant Numbers
conceived and conceptualised the study; B.P.M.N.
designed the bioinformatic analyses; M.W.S
performed the bioinformatic analyses; B.P.M.N
performed the vector construction with assistance from Y.K
Download citation
DOI: https://doi.org/10.1038/s41467-023-44408-3
Metrics details
A Correction to this article was published on 04 October 2021
This article has been updated
Malaria remains a serious public health problem in Cameroon
Implementation of control interventions requires prior knowledge of the local epidemiological situation
Here we report the results of epidemiological and entomological surveys carried out in Tibati
an area where malaria transmission is seasonal
6 years after the introduction of long-lasting insecticidal bed nets
Cross-sectional studies were carried out in July 2015 and 2017 in Tibati
Thick blood smears and dried blood spots were collected from asymptomatic and symptomatic individuals in the community and at health centers
and used for the molecular diagnosis of Plasmodium species
Adult mosquitoes were collected by indoor residual spraying and identified morphologically and molecularly
and positivity of PCR-positive samples was confirmed by Sanger sequencing
Overall malaria prevalence in our study population was 55.0% (752/1367) and Plasmodium falciparum was the most prevalent parasite species (94.3%)
ovale (0.8%); 92 (12.7%) infections were mixed infections
Infection parameters varied according to clinical status (symptomatic/asymptomatic) and age of the sampled population and the collection sites
Infection prevalence was higher in asymptomatic carriers (60.8%)
but asexual and sexual parasite densities were lower
Prevalence and intensity of infection decreased with age in both the symptomatic and asymptomatic groups
Heterogeneity in infections was observed at the neighborhood level
Among the 592 Anopheles mosquitoes collected
A total of 26 (4.39%) mosquito specimens were infected by Plasmodium sp
and the three Anopheles mosquitoes transmitted Plasmodium at equal efficiency
coluzzii specimen infected by Plasmodium vivax
which confirms circulation of this species in Cameroon
The positivity of all 26 PCR-positive Plasmodium-infected mosquitoes was successively confirmed by sequencing analysis
Our study presents the baseline malaria parasite burden in Tibati
Our results highlight the high malaria endemicity in the area
and hotspots of disease transmission are identified
Parasitological indices suggest low bednet usage and that implementation of control interventions in the area is needed to reduce malaria burden
We also report for the first time a mosquito vector with naturally acquired P
vivax with human infections in both Duffy-positive and -negative individuals has been reported
no study has yet looked at which local malaria vector species is involved in the transmission of P
we aimed to characterize malaria parasites and vectors circulating in an area of seasonal transmission in the Adamawa region
vivax detection as this species was recently reported in other parts of the country
and we conducted entomological surveys to identify its putative vector
which has not been investigated in earlier studies
Our main goal was to examine the potential risk of transmission of P
vivax and to determine which local malaria vector species could be implicated in its transmission in this region of Cameroon
Map of Tibati showing collection sites
Blood samples were collected from symptomatic malaria patients in three health facilities: the Missionary hospital of Ngaoubela (MDH)
District Medical Center (DMC) and the Integrated Medical Center (IMC)
Blood samples were collected from asymptomatic persons in the community in eight neighborhoods of Tibati: Malarba
which were the same neighborhoods that mosquito collections were performed
stored at room temperature in a desiccant container and brought to the Malaria Research Unit for further analysis
Plasmodium-positive patients were treated with an ACT according to the recommendations of the Ministry of Health of Cameroon (National Malaria Control Program [NMCP])
All female Anopheles specimens were dissected individually
and the carcasses and head-thoraces were stored separately in tubes containing a desiccant
archived and kept at − 20 °C for further molecular analysis
three 5-mm discs of a dried blood spot were cut out using a sterile paper punch and transferred to a 1.5-ml sample tube filled with 500 µl of sterile water
The tubes were vortexed three times for at least 5 s each time and centrifuged at 2000 rpm for few seconds
the paper discs were transferred into a 0.5-ml tube containing 120 µl sterile water and incubated at 95 °C in a heat block for 15 min (Gene Amp®; Applied Biosystems
the tubes were centrifuged for few seconds
and the extracted DNA was stored at − 20 °C for molecular diagnosis
To optimize the DNA extraction and allow for a better separation of DNA from some nucleases
we used the Qiagen DNeasy® Blood and Tissue Kit (Hilden
Germany) to isolate DNA from the blood samples collected in 2017
in accordance with the manufacturer’s instructions
and resuspended the isolated DNA in 60 µl elution buffer
DNA from the head-thoraces was isolated using the Qiagen DNeasy® Blood and Tissue Kit according to the manufacturer’s instructions
The amplified products were visualized by electrophoresis in a 2% agarose gel and stained with SYBR dye (Green Nucleic Acid stain; Biotium
The expected amplicon size was 276 bp for P
All PCR products were analyzed in a 2% agarose gel
Reaction mixtures were performed with 1 μl of template DNA in a final volume of 10 μl with EvaGreen® (5× HOT Pol EvaGreen® RT PCR Mix Plus; Euromodex
France) and amplified in a 7300 Real-time PCR system (Applied Biosystems)
A dissociation curve was used to estimate the specific melting temperature for each reaction
The final volume of the reaction mixture was 10 μl
The PCR conditions consisted of an initial preincubation step at 95 °C for 10 min; followed a three-step amplification of 95 °C/10 s (ramp 4.4 °C/s)
50 °C/5 s (2.2 °C/s) and 72 °C/20 s (4.4 °C/s)
Amplification was directly followed by a melting program of 95 °C/120 s (2.2 °C/s)
68 °C/120 s (2.2 °C/s) and 90 °C/1 s (ramp 0.2 °C/s with 15 readings/°C)
and a stepwise temperature increase of 0.03 °C/s until 95 °C
falciparum 3D7 clone was used as positive control for Plasmodium species differentiation
The qPCR products of Plasmodium-positive mosquitoes were sequenced using the Big Dye Terminator v3.1 Sequencing Kit (Applied Biosystems) and run on an Applied Biosystems 3130xl Sequencer at the GenSeq technical facility of the Institut des Sciences de l’Evolution de Montpellier
Sequences were verified using SeqScape software (Applied Biosystems)
Data were stored in Microsoft Office Excel files (Microsoft Corp.
USA) and transferred into GraphPad Prism 7 (GraphPad Software Inc.
The Mann–Whitney test was used to compare mean parasite densities according to clinical status
The Chi-square test was used to compare the prevalence of Plasmodium infections among population age groups and mosquito species
The non-parametric Kruskal–Wallis test was used to assess differences in parasite densities between age groups
The significance threshold was set at alpha = 0.05
The relative risk (RR) was computed to estimate the protection associated with LLIN use
R software version 3.5.3 with maptools and ggplot2 packages was used to generate the map and figures
respectively (R Foundation for Statistical Computing
while melting temperatures of qPCR-positive controls were retrieved by using functional bases (without package)
A total of 1367 participants were enrolled in this study, 418 through the health centers (symptomatics) and 949 at the community level (asymptomatics). The main characteristics of the participants are detailed in Table 1
Boxplot of parasite densities for each age group in symptomatic and asymptomatic individuals
malaria diagnostic tools revealed 752 (55.01%) infections with Plasmodium
The infection rate was 41.8% (175/418) in samples from symptomatic patients and 60.8% (577/949) in samples from asymptomatic individuals
which is significantly different (X2 = 41.21; P < 0.0001)
Plasmodium falciparum was the most prevalent Plasmodium species collected during both collection periods and represented 98.8 and 92.9% of human infections in 2015 and 2017
ovale each accounted for 0.6% of clinical infections
in samples from asymptomatic persons a higher prevalence was recorded for P
Mixed infections were only found in asymptomatic carriers and were mostly represented by P
vivax infection was detected in any blood sample
LLIN use based on self-report only conferred slight protection: malaria infection among LLIN users was only 7% less than that among non-users (non-significant difference; RR = 0.89
95% confidence interval [CI] 0.58–0.98); however
the difference varied with collection site
with up to 20% protection found in Malarba
Distribution of Anopheles mosquitoes according to collection time
Melting curve peak of the YK60 and NG239 samples identified as being infected by a Plasmodium sp
The x-axis represents the melting temperature (Tm) and the y-axis represents the results of quantitative PCR on the Lightcycler real-time PCR system (dF/dT negative derivative of the fluorescence/derivative temperature)
Each red and blue curve represents the Tm of each Plasmodium falciparum-positive control
The green curve represents the Tm of the sample
All Plasmodium-positive specimens were monoinfected
A single sample was found by qPCR to be positive for An
All Plasmodium-positive mosquitoes were processed for sequencing, and the sequence alignment of the P. vivax infection is presented in Fig. 5.
vivax sample isolated from an infected Anopheles coluzzii sampled in 2015
with the reference DNA sequence Pv_ECPR (accession number AY423071.1)
P4G2_F and P4G2_R represent the forward and reverse sequences
Numbering corresponds to that of the Pv_ECPR sequence
This study was performed to assess epidemiological and entomological parameters of malaria in Tibati
a locality situated in the Adamawa region of Cameroon
9% of the negative blood smears by microscopic examination were found to be positive upon molecular analysis in our study
and modifications to the habitat in Tibati (e.g
home improvement) that occurred during the 2 years between the studies have probably contributed to the creation of breeding sites more suitable for An
which is a widely used method for sampling endophilic mosquitoes
could have favored the collection of endophilic resting Anopheles over other mosquito species
The percentage of infected mosquitoes sampled in 2017 varied between neighborhoods
with the highest infection rate recorded in Yoko (5.2%)
which could be explained by a high malaria prevalence and the lower LLIN coverage (21.1%) in this neighborhood
Plasmodium vivax can then be considered to circulate in the study area and may also be underestimated
vivax infections in humans are “hidden” since hypnozoites lie dormant in the liver for several months (or years) where they are undetected
A limitation to this study is that mosquito samples were obtained at a single collection time and
the results only provide a one-shot picture of malaria transmission in the study site
Longitudinal surveys would be necessary to follow the dynamics of malaria transmission and
will be crucial to perform parasitological and entomological surveys before and after the implementation of control interventions
A second limitation is that sample sizes were small
which is possibly the reason we did not detect P
Regular monitoring of malaria infections will be necessary to assess the true circulation of P
vivax in the area; we cannot exclude the possibility that the P
vivax-positive mosquito got infected while feeding on a non-resident as Tibati is a cross-border city with a high circulation of people
We have provided a picture of the epidemiological and entomological malaria situation in Tibati
a small town in the Adamawa region of Cameroon
Malaria prevalence varied from 42% in symptomatic patients to 61% in asymptomatic individuals
and this finding highlights the high malaria endemicity in the area
were responsible for the transmission of the disease
with all three species contributing equally to Plasmodium transmission
Parasitological indices suggest low bednet usage and that the implementation of control interventions in the area is needed to reduce the malaria burden
We identified hotspots of disease transmission
and these sites should be the target of malaria control efforts
coluzzii mosquito prompts for regular monitoring as the spread of this species could introduce further complexity into malaria epidemiology and control measures in this area
All data generated or analyzed during the current study are included in this published article
A Correction to this paper has been published: https://doi.org/10.1186/s13071-021-04912-1
PCR‐restriction fragment length polymorphism
World Health Organization. World malaria report. Geneva: World Health Organization; 2020. https://www.who.int/publications/i/item/9789240015791
Complexity of the malaria vectorial system in cameroon: contribution of secondary vectors to malaria transmission
Anopheles species of the Mount Cameroon region: biting habits
feeding behaviour and entomological inoculation rates
Systematique et biologie des Anopheles vecteurs de Plasmodium en Afrique Données récentes
Unravelling complexities in human malaria transmission dynamics in Africa through a comprehensive knowledge of vector populations
Short report: first report of knockdown mutation in the malaria vector Anopheles gambiae from Cameroon
Anopheles coluzzii larval habitat and insecticide resistance in the island area of Manoka
Multiple insecticide resistance mechanisms in Anopheles gambiae s.l
Multiple insecticide resistance in the malaria vector Anopheles funestus from Northern Cameroon is mediated by metabolic resistance alongside potential target site insensitivity mutations
The distribution of insecticide resistance in Anopheles gambiae s.l
Patterns of anopheline feeding/resting behaviour and Plasmodium infections in North Cameroon
2011–2014: implications for malaria control
Status of insecticide resistance and its mechanisms in Anopheles gambiae and Anopheles coluzzii populations from forest settings in South Cameroon
Molecular monitoring of Plasmodium falciparum drug susceptibility at the time of the introduction of artemisinin-based combination therapy in Yaoundé
Prevalence of Plasmodium falciparum parasites resistant to sulfadoxine/pyrimethamine in pregnant women in Yaoundé
Cameroon: emergence of highly resistant pfdhfr/pfdhps alleles
Molecular markers for artemisinin and partner drug resistance in natural Plasmodium falciparum populations following increased insecticide treated net coverage along the slope of mount Cameroon: cross-sectional study
Effects of drug policy changes on evolution of molecular markers of Plasmodium falciparum resistance to chloroquine
and sulphadoxine-pyrimethamine in the south west region of Cameroon
sulphadoxine-pyrimethamine and their combination for the treatment of uncomplicated Plasmodium falciparum malaria in children in Cameroon at the time of policy change to artemisinin-based combination therapy
Genetic diversity of Plasmodium falciparum in Bolifamba
on the slopes of Mount Cameroon: influence of MSP1 allelic variants on symptomatic malaria and anaemia
Genetic diversity of Plasmodium falciparum and genetic profile in children affected by uncomplicated malaria in Cameroon
Identification of the Plasmodium species in clinical samples from children residing in five epidemiological strata of malaria in Cameroon
Comparison of the accuracy of four malaria diagnostic methods in a high transmission setting in coastal Cameroon
Epidemiology of malaria in three geo-ecological zones along the chad-cameroon pipeline
Rapport d’activité 2012 du programme de lutte contre le paludisme
Molecular evidence of Plasmodium vivax mono and mixed malaria parasite infections in duffy-negative native Cameroonians
Molecular typing reveals substantial Plasmodium vivax infection in asymptomatic adults in a rural area of Cameroon
Molecular evidence of Plasmodium vivax infection in Duffy negative symptomatic individuals from Dschang
An additional observation of Plasmodium vivax malaria infection in Duffy-negative individuals from Cameroon
World Health Organization. World Malaria Report. Geneva; 2019. https://www.who.int/publications/i/item/9789241565721
The duffy blood groups of New York negroes: the phenotype Fy(a−b−)
The resistance factor to Plasmodium vivax in blacks
Du¡y-null promoter heterozygosity reduces DARC expression and abrogates adhesion of the P
vivax ligand required for blood-stage infection
Plasmodium vivax in Africa: hidden in plain sight
Transmission of Plasmodium vivax in Duffy-negative individuals in central Sudan
Widespread distribution of Plasmodium vivax malaria in Mauritania on the interface of the Maghreb and West Africa
Microscopic and molecular evidence of the presence of asymptomatic Plasmodium falciparum and Plasmodium vivax infections in an area with low
seasonal and unstable malaria transmission in Ethiopia
Multiplicity and molecular epidemiology of Plasmodium vivax and Plasmodium falciparum infections in East Africa
Duffy negative antigen is no longer a barrier to Plasmodium vivax–molecular evidences from the African West Coast (Angola and Equatorial Guinea)
Plasmodium vivax Infections over 3 Years in Duffy blood group negative Malians in Bandiagara
Evidence for the transmission of Plasmodium vivax in the republic of the Congo West Central Africa
The hide and seek of Plasmodium vivax in West Africa: report from a large-scale study in Beninese asymptomatic subjects
Molecular identification of Plasmodium species responsible for malaria reveals Plasmodium vivax isolates in Duffy negative individuals from southwestern Nigeria
PNDP. Plan communal de développement de la commune de Tibati. 2003. https://www.pndp.org/plan-communaux-developpement.php?dest=plan&crc=23
Distribution of Plasmodium falciparum gametocytes and malaria-attributable fraction of fever episodes along an altitudinal transect in Western Cameroon
A Supplement to the Anophelinae of Africa South of the Sahara
The Anophelinae of Africa south of the Sahara (Ethiopian Zoogeographical Region)
Johannesburg: South African Institute for Medical Research
A reliable and effective method of DNA isolation from old human blood paper cards
A ribosomal RNA gene probe differentiates member species of the Anopheles gambiae complex
multiplex PCR to detect malarial parasites and identify the species present
Simultaneous identification of species and molecular forms of the Anopheles gambiae complex by PCR-RFLP
A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group
Application of a qPCR assay in the investigation of susceptibility to malaria infection of the M and S Molecular Forms of An
Evaluation of a real-time PCR assay for malaria diagnosis in patients from Vietnam and in returned travellers
Real-Time PCR for detection and identification of Plasmodium spp
Evaluating a malaria intervention strategy using knowledge
practices and coverage surveys in rural Bolifamba
and polymerase chain reaction for accurate diagnosis of Plasmodium falciparum malaria
Predominance of Plasmodium malariae-falciparum Co-infection by molecular speciation in Bangolan
Falciparum malaria as an emerging cause of fever in adults living in Gabon Central Africa
Concurrent Chikungunya and dengue virus infections during simultaneous outbreaks
A survey of malaria and some arboviral infections among suspected febrile patients visiting a health centre in Simawa
Seroprevalence and distribution of flaviviridae
and bunyaviridae arboviral infections in rural cameroonian adults
Programme National de Lutte contre le Paludisme
Rapport d’activités du programme national de lutte contre le paludisme
Asymptomatic Plasmodium falciparum malaria prevalence among adolescents and adults in Malawi
Reported reasons for not using a mosquito net when one is available: a review of the published literature
attitudes and prevention of malaria in the cities of Douala and Yaoundé (Cameroon)
Asymptomatic malaria infections: detectability
transmissibility and public health relevance
The silent threat: asymptomatic parasitemia and malaria transmission
Plasmodium falciparum mating patterns and mosquito infectivity of natural isolates of gametocytes
Parasite threshold associated with clinical malaria in areas of different transmission intensities in north eastern Tanzania
Attributable fraction estimates and case definitions for malaria in endemic areas
Mortality and morbidity from malaria among children in a rural area of The Gambia
Effect of transmission intensity on hotspots and micro-epidemiology of malaria in sub-Saharan Africa
Do hotspots fuel malaria transmission: a village-scale spatio-temporal analysis of a 2-year cohort study in The Gambia
Identification of hot spots of malaria transmission for targeted malaria control
The impact of hotspot-targeted interventions on malaria transmission in Rachuonyo South District in the Western Kenyan Highlands: a cluster-randomized controlled trial
Analysis of genetic diversity in the chloroquine-resistant gene Pfcrt in field Plasmodium falciparum isolates from five regions of the southern Cameroon
Impact of exposure to mosquito transmission-blocking antibodies on Plasmodium falciparum population genetic structure
Imported Plasmodium vivax malaria in France: geographical origin and report of an atypical case acquired in Central or Western Africa
Case report: an unusual late relapse of Plasmodium vivax malaria
Growing evidence of Plasmodium vivax across malaria-endemic Africa
1910) and other local anophelines in human malaria transmission in the northern savannah of Cameroon: a cross-sectional survey
Habitat suitability and ecological niche profile of major malaria vectors in Cameroon
Malaria vectors and transmission dynamics in coastal south-western Cameroon
Bionomics and insecticides resistance profiling of malaria vectors at a selected site for experimental hut trials in central Cameroon
Review of the evolution of insecticide resistance in main malaria vectors in Cameroon from 1990 to 2017
Investigation of the influence of a glutathione S-transferase metabolic resistance to pyrethroids/DDT on mating competitiveness in males of the African malaria vector Anopheles funestus
Gene flow between chromosomal forms of the malaria vector Anopheles funestus in Cameroon
Anopheles gambiae distribution and insecticide resistance in the cities of Douala and Yaoundé (Cameroon): influence of urban agriculture and pollution
Distribution des espèces et de la fréquence du gène Kdr chez les populations d’Anopheles gambiae s.s
et d’Anopheles coluzzii dans cinq sites agricoles de la Côte d’Ivoire
Anthropogenic habitat disturbance and ecological divergence between incipient species of the malaria mosquito Anopheles gambiae
Ecological niche partitioning between Anopheles gambiae molecular forms in Cameroon: the ecological side of speciation
Water quality and Anopheles gambiae larval tolerance to pyrethroids in the cities of douala and yaoundé (Cameroon)
Malaria in Mauritania: retrospective and prospective overview
Small-scale land-use variability affects Anopheles spp
distribution and concomitant Plasmodium infection in humans and mosquito vectors in southeastern Madagascar
Malaria transmission and vector biology in manarintsoa
Blood meal sources and entomological inoculation rates of anophelines along a highland altitudinal transect in south-central Ethiopia
Moderate transmission but high prevalence of malaria in Madagascar
Evidence for transmission of Plasmodium vivax among a duffy antigen negative population in western Kenya
Download references
We are grateful to all the residents of Tibati who agreed to participate in this study and gave permission to enter their houses for mosquito collection
Our appreciation is also extended to Estelle Esssangui and Balotin Fongang for their support and cooperation during data analysis
Our sincere thanks go to Rhoel Dinglasan for his contribution in reviewing the article
This work was carried out as part of an AUF-IRD research project granted to SEN
This research was supported by the French National Research Institute for Sustainable Development (IRD) through the JEAI-IMPALA project granted to SEN
LBFD and EMSM were supported by a doctoral fellowship from the LabEx ParaFrap
Lionel Brice Feufack-Donfack and Elangwe Milo Sarah-Matio contributed equally to this work
Service de Paludisme du Centre Pasteur Cameroun
Isabelle Morlais & Sandrine Eveline Nsango
Institut de Recherche pour le Développement
Christelle Maffo Ngou & Isabelle Morlais
Organisation de Coordination pour la lutte contre les Endémies en Afrique Centrale
Jean-Claude Toto & Parfait Awono-Ambene
Centre for Research in Infectious Diseases
Department of Microbiology and Infectious Diseases
School of Veterinary Medicine and Sciences
Faculté de Médecine et des Sciences Pharmaceutiques de l’Université de Douala (FMSP–UD)
Carole Else Eboumbou Moukoko & Sandrine Eveline Nsango
SEN and IM conceived and designed the study protocol
MMS and SEN carried out the field and laboratory assays
LA and PAA critically reviewed the manuscript
All authors read and approved the final manuscript
Our study was approved by the Cameroon National Ethics Committee (N°2015/04/579/CE/NERSH) and by the Delegate of Public Health from Adamaoua region (N°029/L/RA/DSP/SAGE/BPF/NGD/15;735/L/RA/DSP/SAGE/BPF/NGD/17) and the Health Director of the Evangelical Lutheran Church (N°004/EELC/OSEELC/SRH/15)
All human volunteers were enrolled after providing written informed consent
either by the participants and/or by their legal guardians
Written informed consent were also provided by household owners prior to mosquito collection
Free malaria treatments with ACT were given to all Plasmodium-infected patients as recommended by the Ministry of Health (NMCP)
The authors declare that they have no competing interests
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
unless otherwise stated in a credit line to the data
Download citation
DOI: https://doi.org/10.1186/s13071-021-04745-y
Metrics details
The careHPV assay is a test for high-risk (HR) human papillomaviruses (HPV) detection designed to be affordable in resource-poor settings
We evaluated the performance of careHPV screening among 1052 women living with HIV/AIDS included in the HARP (HPV in Africa Research Partnership) study in Burkina Faso (BF) and South Africa (SA)
Cervical samples were tested for HR-HPV by the careHPV and the INNO-LiPA HPV genotyping Extra assays
visual inspection with acetic acid/Lugol’s iodine (VIA/VILI) and colposcopy
Cervical biopsies were obtained for participants who were HR-HPV DNA positive by careHPV or who had abnormalities detected on cytology
45.1% of women had a positive careHPV test (46.5% in BF
The careHPV positivity rate increased with the grade of cytological lesions
Sensitivity and specificity of careHPV for the diagnosis of CIN2+ (n=60
both countries combined) were 93.3% (95% confidence interval (CI): 83.8–98.2) and 57.9% (95% CI: 54.5–61.2)
careHPV had a similar clinical sensitivity but higher specificity than the INNO-LiPA assay for detection of CIN2+
Our results suggest that careHPV testing is a reliable tool for cervical cancer screening in HIV-1-infected women in sub-Saharan Africa
there is a need for developing preventive measures in this highly exposed population of African women living with HIV/AIDS (WLHA)
cervical cancer screening programs are lacking in most Sub-Saharan African countries and the diagnosis of cervical cancer is generally made at an advanced stage of the disease when treatment is unavailable or ineffective
but evaluation among WLHA have only rarely been conducted
it remained to be demonstrated that careHPV had a good performance among African WLHA
we used the second round of screening in HARP 18 months after enrolment to evaluate the performance of careHPV for the detection of CIN2+
Participants were confirmed HIV-1− seropositive women aged 25–50 years recruited from the HIV outpatient clinic of the University Hospital of Ouagadougou
and HIV treatment centres and surrounding primary health care clinics in Hillbrow
Eligible women were invited for inclusion in the study if they were resident in the recruitment city
did not had a total hysterectomy or history of cervical cancer treatment and were not pregnant or less than 8 weeks post partum
Ethical approval was granted from Ministry of Health in Burkina Faso (no
the Witwatersrand University in South Africa (no
110707) and the London School of Hygiene and Tropical Medicine (no
All women provided a written informed consent at the screening visit
and they were given a reflection period of at least 7 days before enrolment in the study
A second written informed consent was obtained at the enrolment visit for enrolment and follow-up over scheduled visits at months (M) 6
endo- and ectocervical sampling was performed using the careHPV sample collection device consisting of a careBrush and a vial containing 1 ml of careHPV collection medium (Qiagen)
the brush was stirred into the collection medium
the cell collection was homogenised by vortexing and divided into four 0.25-ml aliquots
One aliquot was maintained at 4 °C until careHPV analysis
and the others were cryopreserved at −80 °C
The careHPV test was performed using 50 μl of cervical sample in collection medium according to the manufacturer’s instruction
The tests were performed at the respective sites by medical scientists specifically trained by a Qiagen’s scientist
The positive or negative result of the careHPV assay was displayed by the careHPV test controller without additional specification of the luminescent signal intensity
which is based on PCR amplification of HPV DNA using broad-spectrum SPF10 consensus primers followed by hybridisation of the amplicons with type-specific oligonucleotides probes immobilised on membrane strips
including the 14 HR-HPV types targeted by the careHPV assay
Testing was performed on an aliquot preserved at −80 °C
A sample was considered HPV+ if at least one of the type-specific probes or one of the HPV control probes were detected
An additional cervical brush was collected from the ecto- and endocervix and rolled on a glass slide which was fixed with ethanol for cytological reading using the Papanicolaou method (Pap test)
Conventional cytology was used as liquid-based cytology was not available at that time in the African laboratories involved in the study
All participants had visual inspection with acetic acid/Lugol’s iodine (VIA/VILI) performed by trained nurses and colposcopy performed by trained colposcopists
Systematic four-quadrant biopsy and directed biopsy from any suspicious lesions were performed for participants testing positive by careHPV or who had abnormalities detected on cytology (⩾ASC-US)
The Bethesda system for reporting cervical cytology (Smith, 2002) was used for cytology results and the CIN classification for histology results
Cytological and histological slides were independently examined at each site by two senior pathologists blinded to the other study results
Pathologists were trained before the start of the study in order to harmonise slide interpretation between sites
A quality assessment of over 10% of slides was organised at 6-month intervals by the reference pathology laboratory at Montpellier University Hospital for both sites
in addition to existing internal and external quality assurance schemes adhered to by the National Health Laboratory Service (NHLS) in SA
reviewed all histological slides from women with a local diagnosis of CIN2+ and ∼10% of slides from women with normal or CIN1 histological findings; the final classification of lesions was based on a consensus of the committee
Women included in this analysis comprise all of the HARP participants who were not lost to follow-up at M18 visit
including women who may have been treated for CIN2+ detected at baseline
Proportions were compared between groups using χ2 or Fisher’s exact test
positive and negative predictive values (NPV) were calculated with exact binomial 95% confidence intervals (CI)
separately for each country first and then for both countries combined (sensitivity and specificity only)
sensitivity and specificity analysis to detect CIN2+ was stratified by levels of CD4 T-cell counts (⩽200 cells per mm3
>350 cells per mm3) at entry in the study and at the time of screening (M18)
and by age (<35 and ⩾35 years) and compared across strata using χ2 or Fisher’s exact tests as appropriate
The comparative analysis of performance of all other methods and triage combinations for the detection of CIN2+ is not reported in this paper
Agreement between the careHPV assay and the INNO-LiPA HPV genotyping Extra assay was assessed by percentage overall agreement and prevalence-adjusted bias-adjusted (PABA)-kappa coefficient
All analyses were done using the Stata version 14 software (Stata Corp
Study flowchart.BF=Burkina Faso; SA=South Africa
The detection rates ranged from 40.0 (HPV56) to 86.4% (HPV58)
Cytology results showed that the overall prevalence of high-grade squamous intraepithelial lesions (HSIL) was 10.5% (98/929) with a prevalence of 2.1% (9/426) in BF and of 17.7% (89/503) in SA. As shown in Table 2
the prevalence of HR-HPV detected by careHPV or the INNO-LiPA genotyping Extra assay increased with the lesion grade (P<0.0001)
and the sensitivity and specificity values
for the detection of HSIL were 88.8% (87/98) and 61.8% (514/831) for careHPV
The sensitivity of careHPV for detecting HSIL was lower in SA than in BF (87.6% vs 100%) but specificity was higher (64.7% vs 56.3%)
Negative predictive values for HSIL were 100% (95% CI: 98.4–100.0) and 96.1% (95% CI: 93.1–98.0) in BF and SA
none of these studies had been conducted among WLHA
the possible impact of HIV infection on the performance of careHPV for cervical screening deserved further investigation
The high prevalence of HR-HPV in this highly exposed population
which increases by level of immune suppression
as not all HPV infections will progress to CIN2+ lesions
which would make that HPV assay suitable for screening
The overall sensitivity and specificity of 93.3 and 57.9% observed in the present study indicate that careHPV would perform equally well for cervical cancer screening of WLHA in sub-Saharan Africa
Given the relative low specificity of HPV testing
a triage test such as cytology might be required to determine which women should be referred to colposcopy
The cost-effectiveness of this approach should be evaluated
we can rule out issues of histological misclassification as final histological diagnosis of CIN2+ lesions was established by a consensus Expert Committee reviewing slides from both countries simultaneously
our results indicate that the careHPV test would be a reliable tool for cervical cancer screening in WLHA
Such a cost-affordable test should be considered for implementation in cervical cancer prevention programs in sub-Saharan Africa targeting women living with HIV/AIDS
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms
Jeronimo JA (2015) Performance of cervical cancer techniques in HIV-infected women in Uganda
Castle PE (2014) The concordance of HPV DNA detection by Hybrid Capture 2 and careHPV on clinician- and self-collected specimens
Meijer CJ (2008) Overview of human papillomavirus-based and other novel options for cervical cancer screening in developed and developing countries
Cancer Epidemiol Biomarkers Prev 21: 1434–1438
and cervical neoplasia and cancer in the era of highly active antiretroviral therapy
Franceschi S (2012) Prevalence and determinants of human papillomavirus infection and cervical lesions in HIV-positive women in Kenya
Segondy M (2006) Human papillomavirus genotype distribution and cervical squamous intraepithelial lesions among high-risk women with and without HIV-1 infection in Burkina Faso
Bray F (2015) Cancer incidence and mortality worldwide: sources
methods and major patterns in GLOBOCAN 2012
Smith JS (2013) Validation of cervical cancer screening methods in HIV positive women from Johannesburg South Africa
Smith JS (2010) Association between cervical dysplasia and human papillomavirus in HIV seropositive women from Johannesburg South Africa
Schiffman M (2012) Effectiveness of a simple rapid human papillomavirus DNA test in rural Nigeria
Kiviat NB (2003) Increased risk of high-grade cervical squamous intraepithelial lesions and invasive cancer among African women with human immunodeficiency virus type 1 and 2 infections
Jeronimo J (2014) CareHPV cervical cancer screening demonstration in a rural population of north India
Eur J Obstet Gynecol Reprod Biol 176: 75–79
Qiao YL (2014) A parallel study of careHPV and Hybrid Capture2 human papillomavirus DNA testing for cervical cancer screening in rural China
Longatto-Filho A (2013) Self-collection for high-risk HPV detection in Brazilian women using the careHPV™ test
Segondy M (2015) Comparison of analytical and clinical performances of the Digene HC2 HPV DNA assay and the INNO-LiPA HPV genotyping assay for detecting high-risk HPV infection and cervical neoplasia among HIV-positive African women
Segondy M (2013) Comparison of careHPV and hybrid capture 2 assays for detection of high-risk human Papillomavirus DNA in cervical samples from HIV-1-infected African women
Lorincz AT (2008) A new HPV-DNA test for cervical-cancer screening in developing regions: a cross-sectional study of clinical accuracy in rural China
Meijer CJ (2014) International HPV screening working group
Efficacy of HPV-based screening for prevention of invasive cervical cancer: follow-up of four European randomised controlled trials
Castle PE (2009) Human papillomavirus infection and cervical cytology in HIV-infected and HIV-uninfected Rwandan women
Blumenthal PD (2013) Feasibility of community-based careHPV for cervical cancer prevention in rural Thailand
Download references
Other contributing members of the HARP study group included: E Cutler
We wish to thank the members of the HARP International Scientific Advisory Group (ISAG): C Lacey (University of York
Zimbabwe),Y Qiao (Chinese Academy of Medical Sciences and Peking Union Medical College
China) and S de Sanjosé (Institut Catala d’Oncologia
France) for her participation in the histological classification end point committee
careHPV test kits and testing systems were obtained through a Qiagen Corporation donation programme
The research leading to these results has received funding from the European Commission (EC) 7th Framework Programme under grant agreement No
INSERM U1058 and University Hospital (CHU)
London School of Hygiene and Tropical Medicine
National Institute for Communicable Diseases
Centre de Recherche Biomoléculaire Pietro Annigoni
Centre de Recherche Internationale en Santé
Sinead Delany-Moretlwe & Philippe Mayaud
The authors declare no conflict of interest
This work is published under the standard license to publish agreement
After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/
Download citation
Journal of the Egyptian National Cancer Institute (2023)
Gynecologic Oncology Research and Practice (2017)
SINGAPORE — Even while she was under police investigation for peddling sham investments
using part of her nearly S$300,000 loot to pay off her earlier victims
after she admitted to 10 counts of cheating
Another 21 counts were taken into consideration for sentencing purposes
The court heard that Neo used the S$285,825 she fleeced from nine victims to repay debts she owed to unlicensed moneylenders
and to repay some of the victims from her scams
She has partially repaid S$32,300 — or around 11 per cent of the money she had cheated — to her victims
marketed a sham investment in Janssen beauty products to some of her friends and acquaintances
Deputy Public Prosecutor (DPP) Victoria Ting told the court that Neo claimed each box of these beauty products could be purchased in bulk at S$150 from the United States and sold for S$350
Neo promised her victims monthly payouts that were proportionate to their investments
In one instance between October 2011 and February 2012
59-year-old Wong Ah Sen into investing in the products
promising her that she would get a monthly dividend of S$1,000 for each tranche of S$1,500 invested
who worked at a hair salon beside the beauty salon Neo was employed at
had confided in her about her financial difficulties
Neo said she would help Ms Wong invest the initial S$1,500
In order to raise the funds for the non-existent investment scheme
Mr Khoo then threatened to lodge a police report when the payout was not given at the end of the month
Despite being under police investigation in 2013
Neo tried to sell other beauty products-related investment scams involving deer placenta capsules and facial serums
Neo told one of her clients Tan Chay Hua that the latter could stand to make a S$50 profit for every box of the products sold
The court heard that Neo also misrepresented herself as the boss of a beauty salon and told an acquaintance that she hoped the two could invest in skincare products
defence counsel Wilson Foo argued for a sentence of 30 months’ jail
saying his client suffers from heart problems
Neo’s “amateurish” scams were of “low levels of sophistication”
which cannot be compared with that of someone in a financial position
DPP Ting countered that the fact that Neo was not in a professional financial position was not a mitigating factor
“(Neo’s) experience in the industry would have lent her scams greater credence and believability,” said the prosecutor
That Neo perpetrated more scams even while under police investigation showed that she was “brazen
(and had) a disregard for the law,” DPP Ting added
urging the court to impose a jail sentence of at least three years (36 months)
Neo’s 32-month jail sentence will be backdated to Sept 28
She could have been sentenced to 10 years’ imprisonment
Todayonline.com and Today Online domains and apps are now part of 'Channelnewsasia.com' domain
We know it's a hassle to switch browsers but we want your experience with TODAY to be fast
Upgraded but still having issues? Contact us
Massive MSC Turkiye to Dock in Kribi as New Container Terminal Opens
Kribi Bitumen Plant Set to Start Construction in 2025 with Government Backing
CEMAC Bond Market Hits CFA 8.45 Trillion in March 2025, Interest Rates Drop
Cameroon’s Timber Output Projected to Rise in 2025 Despite Higher Export Taxes
Central Africa Stock Exchange Sees 98% Drop in Trading Value in Q1 2025
Mboa Paris Trains 30 Young Cameroonians to Boost Tech and Entrepreneurship
Cameroon Audit Targets Former Officials for Mismanagement in Agricultural Project
Camwater Seeks Global Bids to Launch Bottled Water Lines in Five Cities
Bafoussam Workshop Highlights Benefits of Cameroon-EU Trade Agreement
Cameroon Could Reach 350,100 Tons of Cotton in 2025 (Beac)
Paul Biya Appoints Johnny Razack as Chair of Cameroon’s National Investment Company
Cameroon Refuses Work Visa Renewal for Casino and Super U Boss Over Toxic Workplace Claims
Cameroon Joins Global Charter to Fight Illegal Fishing
Cameroon Launches Construction of New Gas Filling Center in Kumba
Cameroon Launches New CFA15bn Bond Issue After Recent Success
Cameroon Eases Port Procedures to Keep CFA350bn in Chadian Transit Trade
Douala Doubles Water Storage in Key Districts with New Reservoir
Cameroon’s Supreme Court Reaffirms Exclusive Authority Over Public Finance Management
"11 of the World’s 20 Fastest-Growing Economies in 2025 Are in Sub-Saharan Africa" (Amadou Sy, IMF)
Douala Airport Customs Seizes $2.3mln Worth of Cocaine Bound for India
Douala Port Shares CFA2bn in Rebates with Shippers and Shipping Lines in 2024
New Sosucam CEO Faces Tough Start as Worker Tensions Linger
Rubber Output in Cameroon Could Reach 59,100 Tons in 2025 (BEAC)
Cameroon Could Hit Highest Cocoa Output in Over Five Years
Cameroon and Chad to Inaugurate CFA74bn Cross-Border Bridge on April 28
Cameroon Ranks Last in Global Fiber Optic Development Index
Cameroon’s Finance Ministry Denies Budget Fraud After Audit Flags Irregularities
Mbam Bridge Project in Cameroon Reassigned to CFHEC, Budget Increased
Alucam Faces Financial Crisis: Audit Calls for CFA43bn Recapitalization to Avoid Shutdown
CSPH Begins Construction of Ebolowa Gas Filling Center, Expected to Open in 2027
Cameroon Turns to BEAC Market for CFA25bn as Interest Rates Surge
Every week the economy and investment news from Cameroon
(Business in Cameroon) - The African Research Centre on Bananas and Plantains (CARBAP) is now an international institution since May 30
a statement from the school board said last week in Njombe
It was at the 7th Ordinary Session of the Njombe School
Jean Daniel Ngou Ngoupayou and chaired by Valentin Pangou du Congo
Minister of Scientific Research and Innovation
Authorities see in the new status a valuable tool for advocacy with international partners and the strengthening of partnership with national agricultural research schemes in the region
CARBAP wants to contribute to the development of sustainable agriculture that respects the environment
diversify production and improve the income of farmers through the provision of new resistant varieties
Cameroon Tribune said Thursday June 18 2013
In Central Africa over 8 million tons of plantain and 3 million tons of bananas are produced yearly for a consumer population of 250 million consumers
Publication date 22 April 2014 | 08:15 ICT
chairman of China Railway Major Bridge Engineering Group Co Ltd
general manager of Cambodia Iron & Steel Mining Industry Group
shake hands following a signing ceremony in Phnom Penhin 2012
The China Railway Group’s planned $7.5 billion Cambodian north-south railway line has been delayed due to funding shortages
Originally slated to begin construction last year
told the South China Morning Post that work on the 400-kilometre railway line was on hold due to funding problems
The April 9 the Morning Post’s report did not say when construction would begin
Part of a larger $9.6 billion agreement signed between China Railway and the Chinese-owned Cambodia Iron and Steel Mining Industry Group
the railway is to connect a steel factory to be built in Preah Vihear province in the north to a new port in Koh Kong province on the gulf of Thailand in the south
Construction has yet to begin on any part of what would be Cambodia’s largest-ever infrastructure project and little information has been made available from either of the companies involved or the government since the project was announced on December 31
From the iron-ore mining area of Preah Vihear
the railway is to pass through the provinces of Kampong Thom
Kampong Chhnang and Kampong Speu before reaching the proposed port in Koh Kong
Chinese company Union Development Group holds 45,000 hectares of land concessions in Koh Kong
It would be difficult for the railway to avoid passing through their holding
which includes a golf course and luxury resort
But they say they have not been told of any plans to lay tracks on their turf
“I don’t think they will build the railway through our concession because until today no one has come to contact us,” the company’s representative Ngou Tieng Lung told the Phnom Penh Post in an interview last month
Union Development has not been informed of any progress on the railway by the government
The ministry of public works and transport could not be reached for comment yesterday
told the Post in March that they too have not received word on the project
“I think the company is still in the feasibility study phase because so far I have not received any news from them
If they have any update on the project they have to inform us,” he said
Pheakavanmony attributed the lack of information to delays in China Railway’s feasibility study brought on by political instability from last year’s national election
ADDITIONAL REPORTING BY MAY KUNMAKARA AND DAVID BOYLE
Secretaries General of ministries in Cameroon have said they will contribute to ongoing efforts for the implementation the National Malaria Multisectoral Plan in the country
The administrators met at a working session in Yaounde on 18th July 2024 to discuss a common goal for malaria elimination in Cameroon by 2035
The meeting was a joint initiative of the Ministry of Public Health, the National Malaria Control Programme, and Impact Santé Afrique.
Being the 11th country most affected by Malaria
the implementation of the multisectoral plan is expected to reduce the Malaria burden in the country by 75%
paving the way for its elimination by 2035
“ We must understand that Malaria is a real challenge for our country and this fight must be wholistic
We will try to ensure that our directorate of norms and control is involved in this fight because this is an issue of habitat which is conditioned by key factors such as climate
the fight will not be successful,” Paul Tchawa
Secretary General of the Ministry of the Environment and Sustainable Development said
The National Malaria Multisectoral Plan comes on the heels of the National Plan for the Fight Against Malaria
This plan accounted for the reduction of Cameroon’s Malaria prevalence to 11%
“ Seeing such personalities in such a meeting gives me hope that these ministries are going to be more committed to this fight against Malaria
We want them to tell their Ministers that we need just one percent of their budget to kick out this disease
President of the Parliamentary Caucus for Health Financing said
Cameroon is the 11th country most affected by Malaria
According to recent statistics from the Ministry of Public Health
almost three million cases and nearly 2000 deaths are recorded yearly in Cameroon
Malaria also accounts for about 40% of the causes of hospitalization with children under 5 years of age accounting for 68% of deaths
According to the National Malaria Control Programme
Cameroon’s prevalence should be less than 1%
The structure reports a reduction in mortality rates between 2019 and 2023
unfortunately threatened by a new anopheles mosquito
“You should have noticed that we have lots of cases of Malaria in the urban areas because there are new anopheles mosquitoes which spread the disease quickly when the environment is favorable,” Olivia Ngou Executive Director of Impact Santé Afrique told CRTV WEB
Proper sensitization on malaria prevention techniques in schools
churches and other public places are among issues stakeholders are looking to effectively address
Stakeholders hope the multisectoral Malaria plan will change the tides
and significantly contribute to Malaria elimination in Cameroon
“The multisectoral framework is a real guide to multisectoral actions and resource mobilization developed and validated with the participation and consensus of all stakeholders,” Public Health Minister
“Today we have a strategic document that guides every sector and permits them understand the activities that they can carry out and it will strongly contribute to the reduction of Malaria,” Olivia Ngou
As efforts to implement the plan are being intensified
Manaouda calls on all actors to welcome the new plan and make it a permanent guide for the fight against the disease
Cameroon is the first country in West and Central Africa to have set up a multisectoral plan to fight against Malaria
Malaria stakeholders and other decision makers will put hands on deck to make the plan a reality in Cameroon
They are looking forward to Cameroon’s elimination of Malaria like Cape Verte that just celebrated the elimination of this disease in their country
Les champs obligatoires sont indiqués avec *
mon e-mail et mon site dans le navigateur pour mon prochain commentaire
Δdocument.getElementById( "ak_js_1" ).setAttribute( "value"
Macau picked up its first medal at the Asian Games currently being held in Indonesia
Wushu athlete Huang Junhua won a gold medal in the men’s Nanquan-Nangun category in the Asian Games 2018
During the competition held yesterday morning
Huang scored 9.73 points in the Nangun category
0.03 higher than his earlier score of 9.70 points in the same category
A silver medal went to Quoc Khanh Pham from Vietnam
and a bronze medal was awarded to Yongmun Lee
wushu athlete Harris Horatius failed to secure a medal in the category after he only scored 19.21 points
he managed to improve his score to 9.71 points from an earlier 9.50 points
A total of 28 gold medals were awarded yesterday
more athletes set new records for Macau in swimming
and Chou Kit set a new Macau record of 7 minutes and 51 seconds
although they failed to advance in the competition
You must be logged in to post a comment.
Copyright © Macau Daily Times 2008-2022. All Rights Reserved
A soldier of the Multinational Force of Central Africa searches a man in Bangui. Photo: Reuters
Clashes between Muslims and Christians in Central African Republic’s capital killed three yesterday as angry residents threw grenades and torched homes, witnesses said.
French and African troops deployed in the country have struggled to stop the tit-for-tat violence between Muslim Seleka rebels, who seized power in March, and Christian self-defence militia, clashes that killed more than 1,000 people in December.
Residents of Bangui said that Seleka forces wearing civilian clothes threw grenades at Christian houses in a northern district of the city, setting them alight. Christian youths then launched reprisal attacks, burning nearby Muslim homes.
“The Muslims came and set fire to the houses... Everyone was affected,” said Aristide Yenga, resident of the Ngou Simon neighbourhood in the north of the capital Bangui.
“This morning they began shooting and when we heard that we left for the larger (displaced persons’) camps.”
The body of a Seleka fighter, with his left hand severed off and missing, lay in the middle of a large avenue following the attacks, a Reuters witness said. A resident of the neighbourhood who asked not to be named said that two other Muslims had been killed there overnight.
No peacekeepers were present, although a French helicopterflew overhead, the Reuters witness said. The incidents occurred in the city’s fifth district which houses both Muslims and Christians and near where heavy artillery battles took place the previous two nights.
To view comments, please register for free or log in to your account.
By EMN
1 (EMN): To mark the National Unity Day (Rashtriya Ekta Diwas) mini marathon race was organised by Police department on October 32 in Noklak
Pritpal Kaur said National Unity Day is celebrated every year to mark the birth anniversary of Iron Man of India
Sardar Vallabhbhai Patel who played a major role in the integration of India
To acknowledge his efforts in uniting the nation
India celebrate National Unity Day on his birth anniversary
Kaur advised the students and youths to set aim in life
He also encouraged them to contribute to the society in whatever way possible
He further congratulated all the winners and participants of the mini marathon race and gave away the cash prize and certificates to them
second and third prizes respectively while Monglon S and Mutchiu S got consolidation prizes
second and third winners respectively while Thongkoi T and Tongzen H got consolation prizes
SDO C Phuniang administrated the Rashtriya Ekta Diwas pledge and the programme was chaired by Additional Superintendent of Police Ngou Khiam
The mini marathon race started at Government Middle School Junction (Penshe Sector) to GHSS Noklak (High School B Sector) where the prize distribution ceremony was held
Participants in the unity run comprised of students
The Republic of the Congo has been granted approximately US$ 59M to finance the construction works of the Dolisie-Kibangou road
which is located in the department of Niari (in the southern region of the Central African country)
This is financial facility was granted by the Development Bank of Central African States (BDEAC) subject to an agreement signed mid-August 2021 between Fortunato-Ofa Mbo Nchama
The 93 km long Dolisie-Kibangou road is a part of the Brazzaville-Libreville road which is one of the eleven priority projects of the Regional Economic Program (PER) of the Central African Economic and Monetary Community (CEMAC)
The scope of the Dolisie-Kibangou road project also includes the construction of bridges over the Niari and Louvakou rivers as well as the rehabilitation of 53 km of related rural roads
the rehabilitation of socio-economic infrastructure
the construction of 21 water boreholes along the river axis in Congo and the construction of a forest and wildlife control post in Mila Mila
Also Read: Grand Zambi-Kribi Road construction in Cameroon about 84% complete
In addition to the Dolisie-Kibangou road section
the Brazzaville-Libreville road also includes the construction of the 49 km Ndendé-Doussala section in Gabon and the rehabilitation of the 132 km Ngongo-Kibangou section in Congo
Considered as a strategic road project that will contribute to the integration and development of the Central African sub-region
the Brazzaville-Libreville road is also listed among the priority achievements of the Gabonese government
the latter approved two draft laws relating to borrowing authorizations for a total of US$ 162M for the financing of works related to the construction of the Ndendé-Doussala road section
Welcome to Construction Review a portal that serves the building and construction industry through our blog
social media presence and print publication
Contact us
Guest Posting
Take down policy