A Unique Plasmodium falciparum Kelch 13 Gene Mutation in Northwest Ethiopia

Artemisinin combination therapy (ACT) is the first line to treat uncomplicated Plasmodium falciparum malaria worldwide. Artemisinin treatment failures are on the rise in southeast Asia. Delayed parasite clearance after ACT is associated with mutations of the P. falciparum kelch 13 gene. Patients (N = 148) in five districts of northwest Ethiopia were enrolled in a 28-day ACT trial. We identified a unique kelch 13 mutation (R622I) in 3/125 (2.4%) samples. The three isolates with R622I were from Negade-Bahir and Aykel districts close to the Ethiopia–Sudan border. One of three patients with the mutant strain was parasitemic at day 3; however, all patients cleared parasites by day 28. Correlation between kelch 13 mutations and parasite clearance was not possible due to the low frequency of mutations in this study.A report by World Health Organization (WHO) estimated that 3.3 billion people are at risk of contracting malaria. In 2013 alone, there were an estimated 198 million cases of malaria worldwide with 584,000 deaths. Ninety percent of the deaths occurred in sub-Saharan Africa where children are the main victims.¹ According to the WHO, between 2000 and 2013, an expansion of malaria interventions helped to reduce malaria incidence by 30% globally and by 34% in Africa.¹ Many countries have initiated malaria elimination programs.¹ Artemisinin-based combination therapies (ACTs) have been highly effective first-line drugs for the treatment of uncomplicated malaria worldwide.² However, the recent emergence of Plasmodium falciparum strains that are resistant to artemisinin in southeast Asian countries poses a huge challenge to the effectiveness of ACT.^1,3 The possibility of dissemination to or independent emergence of artemisinin-resistant strains in Africa, where the majority of malaria-associated deaths occur, will have potentially catastrophic outcomes.A recent study using genome sequencing of P. falciparum has demonstrated an association between in vivo delayed parasite clearance, in vitro artemisinin “resistance” in a ring-stage assay, and mutations (Y493H, R539T, I543T, and C580Y) in the propeller domain of the parasite kelch 13 gene located on chromosome 13 (PF3D7_1343700 or PF13_0238).⁴ Moreover, an artemisinin-resistant P. falciparum strain that was selected by dose-escalating in vitro culture harbored the mutation, M476I, in the kelch 13 gene. Another study on samples from different southeast Asian countries confirmed the association of kelch 13 mutations and increased parasite clearance half-life. It also demonstrated the independent emergence of these mutations in different geographical areas.⁵ Targeted genetic engineering of kelch 13 using zinc-finger nuclease technology significantly increases the ring-stage survival rate of the parasite in vitro.⁶ Thus, detection of mutations on kelch 13 can be used to track artemisinin resistance in places where clinical treatment failure has not been observed. Kelch 13 mutation screening studies of P. falciparum isolates collected across sub-Saharan African countries have not identified the genotypes associated with treatment failure. However, several other non-synonymous mutations were detected in these countries.^7,8Ethiopia, with a population of more than 90 million, is a malaria-endemic country situated in sub-Saharan Africa. Approximately 68% of the population of Ethiopia live in malarious areas and are at risk of contracting the disease.⁹ There has been a significant reduction in malaria cases in Ethiopia since the introduction of artemether–lumefantrine for the treatment of uncomplicated falciparum malaria in 2004.^9,10 In this study, we aimed to assess the presence of kelch 13 propeller mutations in P. falciparum isolates from northwest Ethiopia as part of a 28-day ACT clinical trial. We identified a mutation in the P. falciparum kelch 13 gene that has not been observed in Asia and Africa before.The study protocol was reviewed and approved by Research and Ethical Review Committee of School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar (reference no.: SBMLS/344/05). The study was conducted on P. falciparum DNA samples obtained from malaria patients from five different health centers in Amhara Regional State, northwest Ethiopia, between October 2013 and April 2014. The five sites were Addis Zemen, Aykel, Maksegnet, Negade Bahir, and Sanja. A total of 148 confirmed P. falciparum malaria patients were enrolled in a 28-day ACT efficacy trial. Patients were treated with artemether–lumefantrine (IPCA Laboratories, Mumbai, India). A directly observed therapy model was used for the first and each morning dose thereafter. The evening doses were given to the patient/guardian for self-administration in the presence of health extension workers.Peripheral blood was collected by finger prick and thin and thick blood films were prepared on a microscope slide. Giemsa staining and microscopy was performed as described previously.¹¹ Only those with mono-infection with P. falciparum were included in this study. Finger prick blood samples were collected on Whatman 903 filter paper (GE Healthcare, Mississauga, Canada) at the time of diagnosis, air-dried, individually inserted in a zip-lock bag, and transported to University of Calgary, Canada, for molecular analysis. Genomic DNA extraction from the filter paper blood spots was performed as described previously in a study.¹¹ Plasmodium falciparum kelch 13 was amplified from the genomic DNA samples using nested polymerase chain reaction (PCR) with slight modification.⁴ The annealing temperature was set at 65.5°C for 45 seconds for the second step of the nested PCR to increase specificity.^4,12 Sequencing of the kelch 13 propeller domain gene was performed using Applied Biosystems (Burlington, ON, Canada) 3730XL 96 capillary DNA analyzer. Bidirectional kelch 13 sequences from each patient sample and the reference kelch 13 gene sequence (PF3D7_1343700) (Genbank ID: {"type":"entrez-nucleotide","attrs":{"text":"AL844509.2","term_id":"225631926","term_text":"AL844509.2"}}AL844509.2) were aligned using Clustal Omega software (EMBL-EBI, Hinxton, UK). The sequences in the multiple sequences alignment were edited manually by Jalview software (Dundee, UK) to remove the gaps.¹³ Nested PCR was performed to confirm the species of Plasmodium as described previously in a study.¹¹ Modeling of the predicted structure of the mutant kelch 13 propeller protein from Ethiopian isolates and its putative effect on the function of the protein was performed using Phyre2 (London, UK) (PDB ID: 2WOZ).¹⁴A total of 148 confirmed P. falciparum-infected malaria patients (age = 1–69 years) were enrolled in a 28-day ACT treatment trial. The kelch 13 propeller domain gene was amplified and sequenced from 125 of the 148 blood samples collected on filter paper. As shown in Figure 1 shows the predicted model of kelch 13 propeller domain with the R622I mutation. The model predicts that the substitution from basic-to-aliphatic residue at this position is likely to impact the protein function. Confirmation of the species of the parasite in the samples where mutation was detected was performed using nested PCR to amplify the small subunit ribosomal RNA gene.¹⁵ Patients with mutant kelch 13 were infected with P. falciparum only.Open in a separate windowFigure 1.Predicted model of the Plasmodium falciparum kelch 13 propeller protein with the R622I mutation. (A) Location of the mutation sites relative to the overall model of the protein. The beta-propeller domain of the btb-kelch protein Krp1 (PDB ID: 2WOZ) was used as the modeling template. (B) The colored bars indicate the probability that a mutation to the corresponding residue will have some effect on function of the protein (mutation sensitivity) or on the phenotype of the organism. The tall and red bars indicate that the likelihood is high. Short and blue bars indicate that the likelihood is low. The R622I substitution (vertical arrow) is predicted to impact protein function.

Table 1

Plasmodium falciparum kelch 13 gene nested PCR and mutation results from study sites in northwest Ethiopia

Thermo-Physiological Comfort Properties of Sportswear with Different Combination of Inner and Outer Layers

Consumers expect high-performance functionality from sportswear. To meet athletic and leisure-time activity requirements, further research needs to be carried out. Sportswear layers and their specific thermal qualities, as well as the set and air layer between materials, are all important factors in sports clothing. This research aims to examine the thermal properties of sports fabrics, and how they are affected by structure parameters and maintained with different layers. Three inner and four outer layers of fabric were used to make 12 sets of sportswear in this study. Before the combination of outer and inner layers, thermal properties were measured for each individual layer. Finally, the thermal resistance, thermal conductivity, thermal absorptivity, peak heat flow density ratio, stationary heat flow density, and water vapor permeability of bi-layered sportswear were evaluated and analyzed. The findings show that sportswear made from a 60% cotton/30% polyester/10% elastane inner layer and a 100% polyester outer layer had the maximum thermal resistance of 61.16 (×10³ K·m² W⁻¹). This performance was followed by the sample made from a 90% polyester/10% elastane inner layer and a 100% polyester outer layer, and the sample composed of a 100% elastane inner layer and a 100% polyester outer layer, which achieved a thermal resistance value of 60.41 and 59.41 (×10³ K·m² W⁻¹), respectively. These results can be explained by the fact that thicker textiles have a higher thermal resistance. This high-thermal-resistance sportswear fabric is appropriate for the winter season. Sportswear with a 90% polyester/10% elastane inner layer had worse water vapor resistance than sportswear with a 60% cotton/30% polyester/10% elastane and a 100% elastane layer. Therefore, these sports clothes have a higher breathability and can provide the wearers with very good comfort. According to the findings, water vapor permeability of bi-layered sportswear is influenced by geometric characteristics and material properties.     

Predictors of Mortality among Adult People Living with HIV and Its Implications for Appointment Spacing Model Approach Care

BackgroundEthiopia has been motivated to achieve a goal of “appointment spacing model approach care.” However, little has been documented on the predictor of mortality and challenges of sustainable HIV care. Therefore, the aim of this study was to determine predictors of mortality among adult people living with HIV/AIDS on antiretroviral therapy (ART).MethodsA retrospective cohort study was conducted on 676 adult people living with HIV who enrolled to ART clinic from September 01, 2012 - August 30, 2016. Multivariable Cox Regression analysis was done where adjusted hazard ratio (AHR)with corresponding 95% confidence interval (CI) at P value ≤ 0.05 cut of point was used to identify predictors of mortality.ResultsThe total person-time contributed was 28,209 person-months with an overall mortality incidence rate of 11 per 1000 person-months observation. The cumulative mortality incidence among females over the study period was 16.8% (64/382). Severe undernourishment and moderate malnutrition at baseline, younger age, female sex, single, divorced, illiterate, lack of disclosure, advanced WHO clinical stage, seeking treatment outside catchment area, rural residence and immunological failure were found to be independent predictors of mortality.ConclusionsPoor nutritional status at baseline, advanced stage of HIV disease, occurrence of treatment failure, female sex, substance abuse, lack of social support, immunological failure, clinical failure, and younger age, low level of education and poor physical access to healthcare facility were found to be important predictors of mortality. Intervening, those factors as routine and part of the appointment spacing model care can improve survival of PLWHA.     

Prevalence and associated factors of female genital mutilation among Somali refugees in eastern Ethiopia: a cross-sectional study

Background  

Eastern Ethiopia hosts a substantial number of refugees originated from Somalia. Female genital mutilation (FGM) is a common practice in the area, despite the campaigns to eliminate it.     

Review on the Integration of Microelectronics for E-Textile

Modern electronic textiles are moving towards flexible wearable textiles, so-called e-textiles that have micro-electronic elements embedded onto the textile fabric that can be used for varied classes of functionalities. There are different methods of integrating rigid microelectronic components into/onto textiles for the development of smart textiles, which include, but are not limited to, physical, mechanical, and chemical approaches. The integration systems must satisfy being flexible, lightweight, stretchable, and washable to offer a superior usability, comfortability, and non-intrusiveness. Furthermore, the resulting wearable garment needs to be breathable. In this review work, three levels of integration of the microelectronics into/onto the textile structures are discussed, the textile-adapted, the textile-integrated, and the textile-based integration. The textile-integrated and the textile-adapted e-textiles have failed to efficiently meet being flexible and washable. To overcome the above problems, researchers studied the integration of microelectronics into/onto textile at fiber or yarn level applying various mechanisms. Hence, a new method of integration, textile-based, has risen to the challenge due to the flexibility and washability advantages of the ultimate product. In general, the aim of this review is to provide a complete overview of the different interconnection methods of electronic components into/onto textile substrate.     

Prevalence of Chlamydia specific antibodies among obstetric and gynecological outpatients in Gondar,north-west Ethiopia

A total of 533 obstetric and gynecological outpatients who visited the dedicated clinics in Gondar, Ethiopia, between April and August 1995 were investigated for Chlamydia-specific antibodies. Relevant epidemiological and clinical data were collected using a pretested questionnaire, clinical examinations and laboratory tests. Earlier, using an antigen specific ELISA technique, we had reported that only 33 (6%) of these women were positive for C. trachomatis in cervical swabs. In this study, we evaluated the prevalence of C. trachomatis-specific antibodies in serum samples of the same patients collected at the time the cervical swabs were taken. Employing a genus-specific ELISA for the detection of chlamydial LPS antibodies, 90% of the sera were found to be positive for IgG, while 49% of the sera contained IgA and 28% IgM antibodies. For 436 of the positive sera, species specificity of the antibodies was determined by means of a micro-immunofluorescence assay. Antibodies against C. trachomatics were present in 75% and antibodies against C. pneumoniae in 69% of these sera. Forty-nine percent of the patients carried antibodies against both species. The antibody based assay estimated that 13.9% of the patients had acute genital chlamydial infection (antigen specific IgM and the simultaneous presence of C. trachomatis specific IgG, IgM and IgA). C. trachomatis antigen detection in cervical swabs apparently underestimated the real frequency of acute infections. Seroprevalence of C. pneumoniae antibodies in the Ethiopian patients was similar to that in industrialized countries.     

Heteronormativity and 'troubled' masculinities among men who have sex with men in Addis Ababa

In most societies, heterosexuality is the dominant way of expressing sexuality and masculinity and those men outside of it are stigmatised and discriminated against. This paper explores the sexual lives of men who have sex with men and the personal and social conflicts that arise as they attempt to both live up to societal expectations and manage their sexual desires. It critically explores how an overriding heteronormativity structures and influences men's perception and understanding of sexuality and masculinity/femininity. The paper draws on data from 24 in-depth/life history interviews, one focus group discussion and ethnographic observation conducted between July 2006 and June 2007. The study reveals that powerful and dominating beliefs about heteronormativity and masculinity result in men who have sex with men dealing with a number of issues of personal conflict and contradiction resulting in uncertainty, resentment, ambivalence, worry and discomfort. Heteronormativity or the expectations of parents, community and society at large is far more influential on the sexuality of men who have sex with men than their own individual desires and needs. The paper concludes that there is little room for individuality for Ethiopian men who have sex with men with their sexual bodies 'belonging' to parents, families and to society at large.