Novel melanin-derived stationary phase for immobilized metal ion affinity chromatography in recombinant His-tagged protein purification.

The matrix of the stationary phase is a crucial element in affinity chromatography for protein purification. Various materials, including polymer or magnetic materials, have been employed as the matrix in the purification of His-tagged protein. Here, for the first time, we utilized a combination of melanin and alginate, both natural polymer materials, to synthesize Ni-melanin/alginate (Ni-M/A) beads for His-tagged protein purification. We investigated the binding of His-tagged Mpro on the Ni-M/A beads, referred to as Ni-M/A-Mpro, and assessed the elution efficiency of Mpro from the beads. Our examination involved FTIR, EDS, XRD, SDS-PAGE, and Western blotting methods. FTIR spectra revealed notable changes in the stretching patterns and intensities of hydroxyl, amine, carbonyl, imine and amide chemical groups, when Mpro protein was present in the Ni-M/A sample. XRD spectra demonstrated the occurrence of two Nickel peaks at 35-40 deg and 40-45 deg in Ni-M/A, but only one nickel peak at 35-40 deg in Ni-M/A-Mpro, indicating the binding of Mpro on the Nickel ions. EDS analysis reported a decrease in the concentration of Nickel on the surface of Ni-M/A from 16% to 7% when Mpro protein was loaded into the stationary phase. Importantly, our data indicated that the purity of the His-tagged protein Mpro after purification reached 97% after just one-step purification using the Ni-M/A stationary phase. Moreover, the binding capacity of Ni-M/A for Mpro was approximately 5.2 mg/g with recovery efficiency of 40%. Our results suggested Ni-M/A as a highly potential solid phase for affinity chromatography in the purification of His-tagged protein.

Treatment of Organic and Sulfate/Sulfide Contaminated Wastewater and Bioelectricity Generation by Sulfate-Reducing Bioreactor Coupling with Sulfide-Oxidizing Fuel Cell.

A wastewater treatment system has been established based on sulfate-reducing and sulfide-oxidizing processes for treating organic wastewater containing high sulfate/sulfide. The influence of COD/SO42- ratio and hydraulic retention time (HRT) on removal efficiencies of sulfate, COD, sulfide and electricity generation was investigated. The continuous operation of the treatment system was carried out for 63 days with the optimum COD/SO42- ratio and HRT. The result showed that the COD and sulfate removal efficiencies were stable, reaching 94.8 ± 0.6 and 93.0 ± 1.3% during the operation. A power density level of 18.0 ± 1.6 mW/m2 was obtained with a sulfide removal efficiency of 93.0 ± 1.2%. However, the sulfide removal efficiency and power density decreased gradually after 45 days. The results from scanning electron microscopy (SEM) with an energy dispersive X-ray (EDX) show that sulfur accumulated on the anode, which could explain the decline in sulfide oxidation and electricity generation. This study provides a promising treatment system to scale up for its actual applications in this type of wastewater.

Effect of Different Catholytes on the Removal of Sulfate/Sulfide and Electricity Generation in Sulfide-Oxidizing Fuel Cell.

Microbial fuel cells are one of the alternative methods that generate green, renewable sources of energy from wastewater. In this study, a new bio-electrochemical system called the sulfide-oxidizing fuel cell (SOFC) is developed for the simultaneous removal of sulfide/sulfide and electricity generation. To improve the application capacity of the SOFC, a system combining sulfate-reducing and sulfide-oxidizing processes for sulfate/sulfide removal and electricity generation was designed. Key factors influencing the sulfide-removal efficiency and electricity-generation capacity of the SOFC are the anolytes and catholytes. The sulfide produced from the sulfate-reducing process is thought to play the key role of an electron mediator (anolyte), which transfers electrons to the electrode to produce electricity. Sulfide can be removed in the anodic chamber of the SOFC when it is oxidized to the element sulfur (S°) through the biochemical reaction at the anode. The performance of wastewater treatment for sulfate/sulfide removal and electricity generation was evaluated by using different catholytes (dissolved oxygen in deionized water, a phosphate buffer, and ferricyanide). The results showed that the sulfate-removal efficiency is 92 ± 1.2% during a 95-day operation. A high sulfide-removal efficiency of 93.5 ± 1.2 and 83.7 ± 2% and power density of 18.5 ± 1.1 and 15.2 ± 1.2 mW/m2 were obtained with ferricyanide and phosphate buffers as the catholyte, respectively, which is about 2.6 and 2.1 times higher than dissolved oxygen being used as a catholyte, respectively. These results indicated that cathode electron acceptors have a direct effect on the performance of the treatment system. The sulfide-removal efficiency and power density of the phosphate buffer SOFC were only slightly less than the ferricyanide SOFC. Therefore, a phosphate buffer could serve as a low-cost and effective pH buffer for practical applications, especially for wastewater treatment. The results presented in this study clearly revealed that the integrated treatment system can be effectively applied for sulfate/sulfide removal and electricity generation simultaneously.

Separation of Rare-Earth Elements from Nitrate Solutions by Solvent Extraction Using Mixtures of Methyltri-n-octylammonium Nitrate and Tri-n-butyl Phosphate.

The article presents data on the solvent extraction separation of rare-earth elements (REEs), such as La(III), Ce(III), Pr(III), and Nd(III), using synergic mixtures of methyltrioctylammonium nitrate (TOMANO3) with tri-n-butyl phosphate (TBP) from weakly acidic nitrate solutions. Specifically, experimental results on separation of REEs, for the pair Ce(III)/Pr(III) for quaternary mixtures of REEs (La(III), Ce(III), Pr(III), Nd(III)) and for the pair La(III)/Pr(III) for solutions containing La(III), Pr(III), and Nd(III), are presented. It was shown that effective separation for the pair Ce(III)/Pr(III) from a solution containing 219 g Ce(III)/L, 106 g La(III)/L, 20 g Pr(III)/L, 55 g Nd(III)/L, and 0.1 mol/L HNO3, was achieved using 56 steps of a multistage, counter-current solvent extraction cascade with scrubbing, at an organic-to-aqueous phase volume ratio (O/A) equal to 2/1 on the extraction section and O/A equal to 4/1 on the scrubbing section, using 3.3 mol/L solutions of the mixture TOMANO3-TBP with molar ratio 0.15:0.85 in dodecane. Separation for the pair La(III)/Pr(III) could be achieved using a solvent extraction cascade with scrubbing in 32 steps at O/A equal to 2/1 on the extraction section and O/A equal to 2.8/1 on the scrubbing section of the solvent extraction cascade from a solution containing 258 g La(III)/L, 58 g Pr(III)/L, 141 g Nd(III)/L, and 0.1 mol/L HNO3 with 3.0 mol/L solution of the mixture TOMANO3-TBP with molar ratio 0.2:0.8 in dodecane.

Weight shapes the intestinal microbiome in preterm infants: results of a prospective observational study.

BACKGROUND: The intestinal microbiome in preterm infants differs markedly from term infants. It is unclear whether the microbiome develops over time according to infant specific factors. METHODS: We analysed (clinical) metadata - to identify the main factors influencing the microbiome composition development - and the first meconium and faecal samples til the 4th week via 16 S rRNA amplican sequencing. RESULTS: We included 41 infants (gestational age 25-30 weeks; birth weight 430-990 g. Birth via Caesarean section (CS) was associated with placental insufficiency during pregnancy and lower BW. In meconium samples and in samples from weeks 2 and 3 the abundance of Escherichia and Bacteroides (maternal faecal representatives) were associated with vaginal delivery while Staphylococcus (skin microbiome representative) was associated with CS. Secondly, irrespective of the week of sampling or the mode of birth, a transition was observed as children children gradually increased in weight from a microbiome dominated by Staphylococcus (Bacilli) towards a microbiome dominated by Enterobacteriaceae (Gammaproteobacteria). CONCLUSIONS: Our data show that the mode of delivery affects the meconium microbiome composition. They also suggest that the weight of the infant at the time of sampling is a better predictor for the stage of progression of the intestinal microbiome development/maturation than postconceptional age as it less confounded by various infant-specific factors.

CAGI 5 splicing challenge: Improved exon skipping and intron retention predictions with MMSplice.

Pathogenic genetic variants often primarily affect splicing. However, it remains difficult to quantitatively predict whether and how genetic variants affect splicing. In 2018, the fifth edition of the Critical Assessment of Genome Interpretation proposed two splicing prediction challenges based on experimental perturbation assays: Vex-seq, assessing exon skipping, and MaPSy, assessing splicing efficiency. We developed a modular modeling framework, MMSplice, the performance of which was among the best on both challenges. Here we provide insights into the modeling assumptions of MMSplice and its individual modules. We furthermore illustrate how MMSplice can be applied in practice for individual genome interpretation, using the MMSplice VEP plugin and the Kipoi variant interpretation plugin, which are directly applicable to VCF files.

Assessing predictions of the impact of variants on splicing in CAGI5.

Precision medicine and sequence-based clinical diagnostics seek to predict disease risk or to identify causative variants from sequencing data. The Critical Assessment of Genome Interpretation (CAGI) is a community experiment consisting of genotype-phenotype prediction challenges; participants build models, undergo assessment, and share key findings. In the past, few CAGI challenges have addressed the impact of sequence variants on splicing. In CAGI5, two challenges (Vex-seq and MaPSY) involved prediction of the effect of variants, primarily single-nucleotide changes, on splicing. Although there are significant differences between these two challenges, both involved prediction of results from high-throughput exon inclusion assays. Here, we discuss the methods used to predict the impact of these variants on splicing, their performance, strengths, and weaknesses, and prospects for predicting the impact of sequence variation on splicing and disease phenotypes.

Assessing evidence for avian-to-human transmission of influenza A/H9N2 virus in rural farming communities in northern Vietnam.

Rural farming communities in northern Vietnam do not routinely practice vaccination for influenza A viruses (IAV) for either humans or poultry, which enables us to study transmission intensity via seroepidemiology. Using samples from a longitudinal cohort of farming households, we determined the number of symptomatic and asymptomatic human infections for seasonal IAV and avian A/H9 over 2 years. As expected, we detected virologically confirmed acute cases of seasonal IAV in humans, as well as large numbers of subclinical seroconversions to A/H1pdm [55/265 (21 %)], A/H3 [95/265 (36 %)] and A/H9 [24/265 (9 %)]. Five of the A/H9 human seroconverters likely represented true infections rather than heterosubtypic immunity, because the individuals seroconverted solely to A/H9. Among co-located poultry, we found significantly higher seroprevalance for A/H5 compared to A/H9 in both chickens and ducks [for northern study sites overall, 337/1105 (30.5 %) seropositive for A/H5 and 123/1105 (11.1 %) seropositive for A/H9].

Comparison of Resting Energy Expenditure Between Cancer Subjects and Healthy Controls: A Meta-Analysis.

There is conflicting evidence surrounding the extent of changes in resting energy expenditure (REE) in cancer. This meta-analysis aimed to establish the mean difference in REE, as kilojoules per kilogram fat-free mass, among cancer patients when compared to healthy control subjects. The secondary aim was to determine differences among different cancer types. PubMed, Cochrane Library, Medline, Science Direct, Scopus, Web of Science, Wiley Online Library, and ProQuest Central were searched from the earliest records until March 2014. Studies were included if measured REE was reported as kilojoules or kilocalories per kilogram fat-free mass (FFM) in adult subjects with cancer. Twenty-seven studies were included in the meta-analysis. Fourteen studies included both cancer (n = 1453) and control (n = 1145) groups. The meta-analysis shows an average increase in REE of 9.66 (95% confidence interval: 3.34, 15.98) kJ/kgFFM/day in cancer patients when compared to control subjects. Heterogeneity was detected (P < 0.001) which suggest variations in REE among cancer types. Elevations are most noticeable in patients with cancers of metabolically demanding organs.

Optimization of sulfide production by an indigenous consortium of sulfate-reducing bacteria for the treatment of lead-contaminated wastewater.

Biological treatment with sulfate-reducing bacteria (SRB) is considered to be an excellent option to remove heavy metals from wastewater. In this study, the optimization of sulfide production for an enhanced removal of lead by a consortium of SRB was carried out based on central composite design and analyzed using response surface methodology (RSM). The sulfide production process was investigated as a function of three independent variables: solution pH (6.5-8.5), lactate concentration (32-96 mM), and sulfate concentration (16-32 mM). RSM analysis showed that the optimum conditions for a high sulfide concentration (14.2 mM) occurred at a pH of 7.5 and at lactate and sulfate concentrations of 53.4 mM and 22.6 mM, respectively. The lead removal efficiency of the SRB consortium using optimum conditions was determined in four parallel anaerobic continuous moving bed biofilm reactors (V = 2 L) that were fed with synthetic wastewater containing dissolved lead at concentrations of 0, 100, 150, 200 mg L(-1) and operated with a hydraulic retention time of 5 days. 99-100 % was removed from synthetic wastewater with lead concentrations of 100 and 150 mg L(-1) during 40 days of operation. For the highest lead concentration of 200 mg L(-1), a decrease in efficiency of removal (96 %) was observed at the end of the experiment.

RNA therapeutics: Molecular mechanisms, and potential clinical translations.

RNA therapies involve the utilization of natural and artificial RNA molecules to control the expression and function of cellular genes and proteins. Initializing from 1990s, RNA therapies now show the rapid growth in the development and application of RNA therapeutics for treating various conditions, especially for undruggable diseases. The outstanding success of recent mRNA vaccines against COVID-19 infection again highlighted the important role of RNA therapies in future medicine. In this review, we will first briefly provide the crucial investigations on RNA therapy, from the first pieces of discovery on RNA molecules to clinical applications of RNA therapeutics. We will then classify the mechanisms of RNA therapeutics from various classes in the treatment of diseases. To emphasize the huge potential of RNA therapies, we also provide the key RNA products that have been on clinical trials or already FDA-approved. With comprehensive knowledge on RNA biology, and the advances in analysis, technology and computer-aid science, RNA therapies can bring a promise to be more expanding to the market in the future.

RNA therapeutics for regenerative medicine.

It is estimated that millions of people around the world experience various types of tissue injuries every year. Regenerative medicine was born and developed for understanding and application with the aim of replacing affected organs or some cells. The research, manufacture, production, and distribution of RNA in cells have acted as a basic foundation for the development and testing of therapies and treatments that are widely applied in different fields of medicine. Vaccines against COVID-19 are considered one of the brilliant and outstanding successes of RNA therapeutics research. With the characteristics of bio-derived RNA therapeutics, the mechanism of rapid implementation, safe production, and flexibility to create proteins depending on actual requirements. Based on the advantages above in this review, we discuss RNA therapeutics for regenerative medicine, and the types of RNA therapies currently being used for regenerative medicine. The relationship between disease and regenerative medicine is currently being studied or tested in RNA therapeutics. We have also covered the mechanisms of action of RNA therapy for regenerative medicine and some of the limitations in our current understanding of the effects of RNA therapy in this area. Additionally, we have also covered developing RNA therapeutics for regenerative medicine, focusing on RNA therapeutics for regenerative medicine. As a final point, we discuss potential applications for therapeutics for regenerative medicine in the future, as well as their mechanisms.

RNA therapeutics for treatment of diabetes.

Diabetes is an ongoing global problem as it affects health of more than 537 million people around the world. Diabetes leaves many serious complications that affect patients and can cause death if not detected and treated promptly. Some of the complications of diabetes include impaired vascular system, increased risk of stroke, neurological diseases that cause pain and numbness, diseases related to the retina leading to blindness, and other complications affecting kidneys, heart failure, muscle weakness, muscle atrophy. All complications of diabetes seriously affect the health of patients. Recently, gene therapy has emerged as a viable treatment strategy for various diseases. DNA and RNA are among the target molecules that can change the structure and function of proteins and are effective methods of treating diseases, especially genetically inherited diseases. RNA therapeutics has attracted deep interest as it has been approved for application in the treatment of functional system disorders such as spinal muscular atrophy, and muscular dystrophy. In this review, we cover the types of RNA therapies considered for treatment of diabetes. In particular, we delve into the mechanism of action of RNA therapies for diabetes, and studies involving testing of these RNA therapies. Finally, we have highlighted the limitations of the current understanding in the mechanism of action of RNA therapies.

Role of DNA methylation in diabetes and obesity.

Due to the fact that the upward trend of several metabolic disorders such as diabetes and obesity, in individuals especially monozygotic twins, who are under the same effects from the environment, are not similar, the role of epigenetic elements like DNA methylation needs taking into account. In this chapter, emerging scientific evidence supporting the strong relationship between changes in DNA methylation and those diseases' development was summarized. Changing in the expression level of diabetes/obesity-related genes through being silenced by methylation can be the underlying mechanism of this phenomenon. Genes with abnormal methylation status are potential biomarkers for early prediction and diagnosis. Moreover, methylation-based molecular targets should be investigated as a new treatment for both T2D and obesity.

Acceptability, Appropriateness, and Feasibility of an Educational Program to Promote Treatment Adherence in Patients with Chronic Obstructive Pulmonary Disease.

The mortality rate due to chronic obstructive pulmonary disease (COPD) has increased annually, and non-adherence to treatment is one reason for this rise. Developing intervention programs to enhance treatment adherence for people with COPD is essential. The purpose of this mixed-methods study was to determine the acceptability, appropriateness, and feasibility of such a program. We sought the opinions of 15 healthcare managers and 15 practicing nurses from three hospitals across Vietnam and conducted group discussions and interviews with 30 patients with COPD. We then formulated integrated conclusions on the acceptability, appropriateness, and feasibility of the program. The overall average score of 12 items to test the acceptability, appropriateness, and feasibility of the program from both healthcare managers and practicing nurses was high (M = 4.31; SD = 0.11) and (M = 4.37; SD = 0.12), respectively. Thirty COPD outpatients agreed the content and plan of the program were necessary for them to enhance their treatment adherence at home. The document content was appropriate, easy to understand, and the support and education provided by nurses was helpful. The educational intervention program to promote treatment adherence for patients with COPD was acceptable, appropriate, and feasible from the views of healthcare managers, nurses, and patients with COPD.

Protection against COVID-19 hospitalisation conferred by primary-series vaccination with AZD1222 in non-boosted individuals: first vaccine effectiveness results of the European COVIDRIVE study and meta-regression analysis.

Background: Vaccine effectiveness (VE) studies with long-term follow-up are needed to understand durability of protection against severe COVID-19 outcomes conferred by primary-series vaccination in individuals not receiving boosters. COVIDRIVE is a European public-private partnership evaluating brand-specific vaccine effectiveness (VE). We report a prespecified interim analysis of primary-series AZD1222 (ChAdOx1 nCoV-19) VE. Methods: Seven Study Contributors in Europe collected data on individuals aged ≥18 years who were hospitalised with severe acute respiratory infection (June 1st, 2021-September 5th, 2022) and eligible for COVID-19 vaccination prior to hospitalisation. In this test-negative case-control study, individuals were defined as test-positive cases or test-negative controls (SARS-CoV-2 RT-PCR) and were either fully vaccinated (two AZD1222 doses, 4-12 weeks apart, completed ≥14 days prior to symptom onset; no booster doses) or unvaccinated (no COVID-19 vaccine prior to hospitalisation). The primary objective was to estimate AZD1222 VE against COVID-19 hospitalisation. A literature review and meta-regression were conducted to contextualise findings on durability of protection. Findings: 761 individuals were included during the 15-month analysis period. Overall AZD1222 VE estimate was 72.8% (95% CI, 53.4-84.1). VE was 93.8% (48.6-99.3) in participants who received second AZD1222 doses ≤8 weeks prior to hospitalisation, with spline-based VE estimates demonstrating protection (VE ≥ 50%) 30 weeks post-second dose. Meta-regression analysis (data from seven publications) showed consistent results, with ≥80% protection against COVID-19 hospitalisation through ∼43 weeks post-second dose, with some degree of waning. Interpretation: Primary-series AZD1222 vaccination confers protection against COVID-19 hospitalisation with enduring levels of VE through ≥6 months. Funding: AstraZeneca.

Early home discharge after robot-assisted coronary artery bypass grafting.

OBJECTIVES: Robot-assisted coronary artery bypass grafting (CABG) has been developed as a less invasive alternative for conventional CABG to enhance postoperative recovery, patient satisfaction and early discharge to home. Furthermore, it may provide a basis for hybrid coronary revascularization. To determine the feasibility of this procedure, we compared robot-assisted with conventional off-pump CABG. METHODS: All consecutive patients undergoing a robot-assisted left internal mammary artery-to-left anterior descending coronary artery procedure were compared to consecutive patients undergoing conventional off-pump CABG for single-vessel disease from October 2016 to July 2019. The primary outcome was discharge to home within 5 days after the operation. Secondary outcomes were total hospital stay, reoperations within 48 h, transfusions, atrial fibrillation, 30-day mortality and quality of life 1 month postoperatively. A propensity matched cohort was assembled to correct for possible confounders. RESULTS: A total of 107 patients who had robot-assisted CABG were compared to 194 patients who had conventional off-pump CABG. The primary outcome was reached in 51% of the robot-assisted group versus 19% of the conventional off-pump group (P < 0.01). The median postoperative hospital stay was 5 days for the robot-assisted group versus 7 days in the conventional off-pump group (P < 0.01). Other secondary outcomes did not differ significantly between the groups, and the quality of life 1 month after the operation was equal. The results after propensity matching were similar. CONCLUSIONS: Early discharge to home is more frequent for patients who have robot-assisted CABG than in those who have conventional off-pump CABG, with no difference in health-related quality of life. Therefore, this approach may reduce healthcare resources and provide a solid basis for hybrid coronary revascularization.

COVID-19 in Southeast Asia: current status and perspectives.

Coronavirus Disease-2019 (COVID-19) has spread globally with catastrophic damages to the public health, social and economy since the beginning of the outbreak. In 2020, Southeast Asia proved that it could prevent the worst effects of a pandemic through the closure of activities and borders and movement restriction, as well as social distancing. Nevertheless, with the occurrence of the common variants of concern (VOCs), especially Alpha (B.1.1.7), Beta (B.1.351), Delta (B.1.617.2), Southeast Asia is facing a significant increase in the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infections. Now, the area also has the threats of the spreading out of the dangerous variant - Omicron (B.1.1.529) from other close countries or regions. COVID-19 countermeasures such as closures and social distancing seem to be insufficient. Moreover, Southeast Asia is being held back by a shortage of vaccines and other medical resources. This work focuses on describing the COVID-19 situation, the virus variants, and the coverage of COVID-19 vaccination in the area. We also provide perspectives on the COVID-19 vaccine distribution, protecting the economic capitals, developing the green zone, and the importance of finding more vaccine supplies in Southeast Asia.

The relationships of nursing students' satisfaction and self-confidence after a simulation-based course with their self-confidence while practicing on real patients in Vietnam

PURPOSE: Simulation teaching refers to the replication of real-life scenarios, enabling students to practice nursing skills and learn actively in a safe environment. It also helps students control their anxiety and fears when caring for real patients. This study investigated the relationships of Vietnamese nursing students' self-confidence in clinical practice with their satisfaction and self-confidence in simulation-based practice. METHODS: This cross-sectional descriptive study included 182 nursing students. The data collection included 2 separate stages with 2 main questionnaires. The Student Satisfaction and Self-Confidence in Learning Scale was used to measure students' satisfaction and self-confidence after learning in the simulation room. The Confidence Scale was used to measure students' self-confidence when first performing techniques on actual patients. Data were analyzed by descriptive and Pearson correlation statistics. RESULTS: Students' satisfaction and self-confidence during the simulation course were quite high (mean±standard deviation [SD], 4.06±0.48 and 4.11±0.46 out of 5.0, respectively). In contrast, their confidence when first practicing on a patient was moderate (mean±SD, 3.19±0.62 out of 5.0). Students' satisfaction showed moderate and weak positive correlations with self-confidence in pre-clinical practice and in clinical practice (r=0.33, P<0.001 and r=0.26, P<0.001, respectively). CONCLUSION: Simulation has become an effective teaching strategy that can help nursing students be well-prepared for clinical placements in Vietnam. An effective nursing education strategy is needed to enhance the satisfaction and self-confidence of nursing students in simulation and then in clinical practice to help achieve professional engagement and development.

Diarrhoea among Children Aged under Five Years and Risk Factors in Informal Settlements: A Cross-Sectional Study in Cape Town, South Africa.

BACKGROUND: There is limited data on the association between diarrhoea among children aged under five years (U5D) and water use, sanitation, hygiene, and socio-economics factors in low-income communities. The study investigated U5D and the associated risk factors in the Zeekoe catchment in Cape Town, South Africa. METHODS: A cross-sectional study was conducted in 707 households in six informal settlements (IS) two formal settlements (FS) (March-June 2017). RESULTS: Most IS households used public taps (74.4%) and shared toilets (93.0%), while FS households used piped water on premises (89.6%) and private toilets (98.3%). IS respondents had higher average hand-washing scores than those of FS (0.04 vs. -0.14, p = 0.02). The overall U5D prevalence was 15.3% (range: 8.6%-24.2%) and was higher in FS than in IS (21.2% vs. 13.4%, respectively, p = 0.01). Water storage >12 h was associated with increasing U5D (OR = 1.88, 95% CI 1.00-3.55, p = 0.05). Water treatment (OR = 0.57, 95%CI 0.34-0.97, p = 0.04), good hand-washing practices (OR = 0.59, 95%CI 0.42-0.82, p = 0.002) and Hepatitis A vaccination (OR = 0.51, 95%CI 0.28-0.9, p = 0.02) had significant preventing effects on U5D. CONCLUSIONS: The study highlights that good hygiene practice is a key intervention against U5D in informal settlements. The promotion of hand-washing, proper water storage, and hygienic breastfeeding is highly recommended.