Distribution of the ACE Gene Polymorphisms in Type 2 Diabetes Mellitus Patients, Their Associations with Nephropathy Biomarkers and Metabolic Indicators at a Tertiary Hospital in Uganda.

Purpose: We aimed at determining the distribution of the ACE insertion/deletion gene polymorphisms among type 2 diabetic patients and their association with the nephropathy biomarkers and the metabolic indicators. Patients and Methods: Data were collected from 237 adult type 2 diabetes mellitus patients receiving healthcare at the diabetic clinic of Mbarara Regional Referral Hospital. Peripheral blood genomic DNA was amplified using a conventional PCR technique and analyzed for the ACE homozygous forms of the insertion (II), deletion (DD) and heterozygous insertion deletion (ID) genotypes as well as their respective allele counts. Biomarkers of nephropathy were analyzed on a Beckman coulter AU480 chemistry analyzer using system compatible reagents. Results: Majority of the participants were older persons (Median = 57, IQR = 49-64) and female 171 (72.2%). Most of them had the Deletion allele 198 (83.5%) and DD genotype 116 (48.9%). At multivariate logistic regression, the nephropathy biomarkers that is microalbuminuria, serum creatinine, urea, eGFR and electrolytes had no association with the ACE I/D alleles or genotypes (p > 0.05). On the other hand, selected metabolic indicators had a positive relationship. The insertion allele was associated with increasing glycated hemoglobin (OR = 1.082, p = 0.019) and decreasing serum glucose levels (OR = 0.891, p = 0.001). Deletion allele was associated with decreasing glycated hemoglobin (OR = 0.924, p = 0.047) and increasing serum glucose levels (OR = 1.208, p = 0.001). ACE II genotype was associated with decreasing serum glucose levels (OR = 0.873, p = 0.029). ACE DD genotype was associated with decreasing glycated hemoglobin (OR = 0.917, p = 0.010) and increasing serum glucose levels (OR = 1.132, p = 0.001). ACE ID genotype was associated with increasing glycated hemoglobin (OR = 1.077, p = 0.022), triglyceride levels (OR = 1.316, p = 0.031) and decreasing serum glucose levels (OR = 0.933, p = 0.038). Conclusion: The presence or absence of the ACE I/D alleles and genotypes affects the ultimate increase or decrease in the serum glucose, glycated hemoglobin and triglyceride levels. Although there was no significant association between the biomarkers of nephropathy and the ACE I/D alleles or genotypes, the above implicated metabolic indicators should be included in healthcare guidelines used when attending to type 2 diabetic patients.

Public health and research ethics education: the experience of developing a new cadre of bioethicists at a Ugandan institution.

Research ethics education is critical to developing a culture of responsible conduct of research. Many countries in sub-Saharan Africa (SSA) have a high burden of infectious diseases like HIV and malaria; some, like Uganda, have recurring outbreaks. Coupled with the increase in non-communicable diseases, researchers have access to large populations to test new medications and vaccines. The need to develop multi-level capacity in research ethics in Uganda is still huge, being compounded by the high burden of disease and challenging public health issues. Only a few institutions in the SSA offer graduate training in research ethics, implying that the proposed ideal of each high-volume research ethics committee having at least one member with in-depth training in ethics is far from reality. Finding best practices for comparable situations and training requirements is challenging because there is currently no "gold standard" for teaching research ethics and little published information on curriculum and implementation strategies. The purpose of this paper is to describe a model of research ethics (RE) education as a track in an existing 2-year Master of Public Health (MPH) to provide training for developing specific applied learning skills to address contemporary and emerging needs for biomedical and public health research in a highly disease-burdened country. We describe our five-year experience in successful implementation of the MPH-RE program by the Mbarara University Research Ethics Education Program at Mbarara University of Science and Technology in southwestern Uganda. We used curriculum materials, applications to the program, post-training and external evaluations, and annual reports for this work. This model can be adapted and used elsewhere in developing countries with similar contexts. Establishing an interface between public health and research ethics requires integration of the two early in the delivery of the MPH-RE program to prevent a disconnect in knowledge between research methods provided by the MPH component of the MPH-RE program and for research in ethics that MPH-RE students are expected to perform for their dissertation. Promoting bioethics education, which is multi-disciplinary, in institutions where it is still "foreign" is challenging and necessitates supportive leadership at all institutional levels.

Trust in Medical Research: A Comparative Study among Patients at a Regional Referral Hospital and Community Members in Lira District, Northern Uganda.

Events such as the Tuskegee syphilis study shaped how the public perceives and trusts medical research globally. However, few studies have examined trust in medical research in developing countries. We tested the hypothesis that levels of trust may be lower among community members compared to hospitalized persons in Uganda. We enrolled 296 participants in rural northern Uganda, and 148(50%) were from the community, 192(65%) were female. Mean level of trust for medical research was higher among hospitalized persons compared to community members (p = 0.0001). Previous research participation (p = 0.03), and willingness to participate in future research (p = 0.001) were positively associated with trust. Medical personnel should engage more with the communities in which they practice fostering trust in medical research.

Estimating cancer incidence in Uganda: a feasibility study for periodic cancer surveillance research in resource limited settings.

BACKGROUND: Population based cancer registries (PBCRs) are accepted as the gold standard for estimating cancer incidence in any population. However, only 15% of the world's population is covered by high quality cancer registries with coverage as low as 1.9% in settings such as Africa. This study was conducted to assess the operational feasibility of estimating cancer incidence using a retrospective "catchment population" approach in Uganda. METHODS: A retrospective population study was conducted in 2018 to identify all newly diagnosed cancer cases between 2013 and 2017 in Mbarara district. Data were extracted from the medical records of health facilities within Mbarara and from national and regional centres that provide cancer care services. Cases were coded according to the International Classification of Diseases for Oncology (ICD-0-03). Data was analysed using CanReg5 and Excel. RESULTS: We sought to collect data from 30 health facilities serving Mbarara district, southwestern Uganda. Twenty-eight sources (93%) provided approval within the set period of two months. Among the twenty-eight sources, two were excluded, as they did not record addresses for cancer cases, leaving 26 sources (87%) valid for data collection. While 13% of the sources charged a fee, ranging from $30 to $100, administrative clearance and approval was at no cost in most (87%) data sources. This study registered 1,258 new cancer cases in Mbarara district. Of the registered cases, 65.4% had a morphologically verified diagnosis indicating relatively good quality of data. The Age-Standardised Incidence Rates for all cancers combined were 109.9 and 91.9 per 100,000 in males and females, respectively. In males, the most commonly diagnosed cancers were prostate, oesophagus, stomach, Kaposi's sarcoma and liver. In females, the most common malignancies were cervix uteri, breast, stomach, liver and ovary. Approximately, 1 in 8 males and 1 in 10 females would develop cancer in Mbarara before the age of 75 years. CONCLUSION: Estimating cancer incidence using a retrospective cohort design and a "catchment population approach" is feasible in Uganda. Periodic studies using this approach are potentially a precious resource for producing quality cancer data in settings where PBCRs are scarce. This could supplement PBCR data to provide a detailed and comprehensive picture of the cancer burden over time, facilitating the direction of cancer control efforts in resource-limited countries.

Comparison of the Nutritional Status of Swiss Albino Mice Fed on Either a Purified or Cereal-Based Diet for 15 weeks.

Background: Laboratory animals are commonly fed on cereal-based diets (CBDs) whose nutrient composition is unknown and may confound the metabolic response to study interventions. Purified diets such as AIN-93M are therefore recommended, as their nutrient composition is known. However, few studies have evaluated their use as adequate control diets. The aim of this study was to compare the nutrition status of Swiss albino mice fed on either CBD or AIN-93M for 15 weeks. Methods: Twenty Swiss albino mice aged 6-8 weeks and weighing 21.7 g ± 0.6 were fed on either CBD or AIN-93M diet for 15 weeks. Their nutritional status was evaluated using anthropometric and hematological indices, serum glucose, total protein, albumin, and total cholesterol to select an appropriate normal control diet. Results: The CBD had low-calorie content (2.57 kcal/g) and protein (11 ± 3.8 g/100 g) compared to AIN-93M (3.8 kcal/g and 14 g/100 g, respectively). The BMI of male mice fed on CBD and AIN-93M diets was significantly higher (P=0.0139 and P=0.0325, respectively) compared to that of females fed on similar diets. Animals in the CBD group had lower hemoglobin (15.1-16.9 g/dl) compared to those in the AIN-93M group (18.1-20.8 g/dl). Serum albumin levels were higher in both male (P=0.001) and female (P=3 × 10-6) mice fed on AIN-93M compared to those fed on CBD. Females in the AIN-93M group had higher cholesterol (P=0.026) than those in the CBD group. Conclusion: The AIN-93 diet of caloric value 3.85 kcal/g (total protein 14 g, total fat 4 g of soy bean oil, fibre 5 g, and total carbohydrate 42 g per 100 g) can be safely used as a normal control diet in long-term research studies using Swiss albino mice.

Microalbuminuria and Traditional Serum Biomarkers of Nephropathy among Diabetic Patients at Mbarara Regional Referral Hospital in South Western Uganda.

BACKGROUND: Diabetic nephropathy (DN) is a common finding in diabetic patients. Microalbuminuria is the earliest clinical evidence of DN. Early detection of microalbuminuria is very important; it allows timely interventions to prevent progression to macroalbuminuria and later end-stage renal disease (ESRD). OBJECTIVES: To determine the prevalence of microalbuminuria in diabetic patients and establish its association with traditional serum renal markers in assessment of incipient nephropathy. METHODS: This cross-sectional study involved 140 participants with diabetes mellitus (DM) attending the diabetic clinic of Mbarara Regional Referral Hospital. Questionnaires were used to obtain participant data after obtaining written informed consent. Data collected included: age, sex, level of education, history of smoking and alcohol consumption, hypertension, body mass index, family history, and duration of DM. Morning spot urine samples were collected from each participant and blood drawn for analysis of other renal markers. Urine microalbumin was determined quantitatively using immunoturbidity assay (Microalbumin kit, Mindray). Serum creatinine and uric acid and glucose levels were determined by spectrophotometric methods. RESULTS: The overall prevalence of microalbuminuria was 22.9%. Using a simple and multiple linear regression model, serum creatinine (ß = 0.010, 95% CI (0.005, 0.014), P = 0.0001) and glucose (ß = 0.030, 95% CI (0.011, 0.048), P = 0.0017) levels were significantly associated with microalbuminuria. After adjusting for linearity, family history of DM was the only predictor of microalbuminuria (ß = 0.275, 95% CI (0.043, 0.508), P = 0.002). Although microalbuminuria was weakly associated with eGFR (OR = 1.2, 95% CI (0.24, 5.96)), the relationship was not statistically significant (P = 0.824). CONCLUSION: The prevalence of microalbuminuria in patients with diabetes in this study was high. The study suggests the need to screen for microalbuminuria early to reduce the possible burden of ESRD. When serum creatinine is used as a renal function marker among diabetic patients, it should be combined with microalbuminuria for better assessment of incipient nephropathy.

Evaluation of the Deki Reader™, an automated RDT reader and data management device, in a household survey setting in low malaria endemic southwestern Uganda.

BACKGROUND: Early diagnosis of suspected malaria cases with a rapid diagnostic test (RDT) has been shown to be an effective malaria control tool used in many resource-constrained settings. However, poor quality control and quality assurance hinder the accurate reporting of malaria diagnoses. Recent use of a portable, battery operated RDT reader (Deki Reader™, Fio Corporation) has shown to have high agreement with visual inspection across diverse health centre settings, however evidence of its feasibility and usability during cross sectional surveys are limited. This study aimed to evaluate the performance of the Deki Reader™ in a cross-sectional survey of children from southwestern Uganda. METHODS: A two-stage, stratified cluster sampling survey was conducted between July and October 2014 in three districts of southwestern Uganda, with varying malaria transmission intensities. A total of 566 children aged 6-59 months were included in the analysis. Blood samples were collected and tested for malaria using: the SD Bioline Malaria Ag Pf/Pan RDT and microscopy. Results were compared between visual inspection of the RDT and by the Deki Reader™. Diagnostic performance of both methods were compared to gold-standard microscopy. RESULTS: The sensitivity and specificity of the Deki Reader™ was 94.1% (95% CI 69.2-99.6%) and 95.6% (95% CI 93.4-97.1%), respectively. The overall percent agreement between the Deki Reader™ and visual RDT inspection was 98.9% (95% CI 93.2-99.8), with kappa statistic of 0.92 (95% CI 0.85-0.98). CONCLUSIONS: The findings from this study suggest that the Deki Reader™ is comparable to visual inspection and performs well in detecting microscopy-positive Plasmodium falciparum cases in a household survey setting. However, the reader's performance was highly dependent on ensuring adequate battery life and a work environment free of dirt particles.

Screening for Glucose-6-Phosphate Dehydrogenase Deficiency Using Three Detection Methods: A Cross-Sectional Survey in Southwestern Uganda.

Despite the potential benefit of primaquine in reducing Plasmodium falciparum transmission and radical cure of Plasmodium vivax and Plasmodium ovale infections, concerns over risk of hemolytic toxicity in individuals with glucose-6-phosphate dehydrogenase deficiency (G6PDd) have hampered its deployment. A cross-sectional survey was conducted in 2014 to assess the G6PDd prevalence among 631 children between 6 and 59 months of age in southwestern Uganda, an area where primaquine may be a promising control measure. G6PDd prevalence was determined using three detection methods: a quantitative G6PD enzyme activity assay (Trinity Biotech® G-6-PDH kit), a qualitative point-of-care test (CareStart™ G6PD rapid diagnostic test [RDT]), and molecular detection of the G6PD A- G202A allele. Qualitative tests were compared with the gold standard quantitative assay. G6PDd prevalence was higher by RDT (8.6%) than by quantitative assay (6.8%), using a < 60% activity threshold. The RDT performed optimally at a < 60% threshold and demonstrated high sensitivity (≥ 90%) and negative predictive values (100%) across three activity thresholds (below 60%, 30%, and 40%). G202A allele frequency was 6.4%, 7.9%, and 6.8% among females, males, and overall, respectively. Notably, over half of the G202A homo-/hemizygous children expressed ≥ 60% enzyme activity. Overall, the CareStart™ G6PD RDT appears to be a viable screening test to accurately identify individuals with enzyme activities below 60%. The low prevalence of G6PDd across all three diagnostic modalities and absence of severe deficiency in our study suggests that there is little barrier to the use of single-dose primaquine in this region.

Asymptomatic Plasmodium Infections in Children in Low Malaria Transmission Setting, Southwestern Uganda(1).

A survey of asymptomatic children in Uganda showed Plasmodium malariae and P. falciparum parasites in 45% and 55% of microscopy-positive samples, respectively. Although 36% of microscopy-positive samples were negative by rapid diagnostic test, 75% showed P. malariae or P. ovale parasites by PCR, indicating that routine diagnostic testing misses many non-P. falciparum malarial infections.

Alcohol exposure among pregnant women in sub-saharan Africa: a systematic review.

BACKGROUND: The prevalence of general alcohol use in many countries of sub-Saharan Africa (SSA) is high. However, research examining alcohol use in among pregnant women within this population is limited. A review of the current status of research examining the prevalence of alcohol exposed pregnancies (AEP) is required to inform future research aiming to decrease this occurrence and its subsequent socio-economic complications. OBJECTIVE: The primary objective was to identify all published papers estimating prevalence and risk-factors of alcohol use among pregnant women in SSA. A secondary objective was to determine changes in alcohol use following pregnancy recognition. METHODS: PubMed/Medline, Embase, IPA, CINAHL were systematically searched using MeSH terms and keywords from inception date to March 2013. Studies from SSA reporting prevalence of alcohol use among pregnant women were included. RESULTS: Twelve studies were identified. Studies varied significantly according to design and study population. Prevalence of alcohol use during pregnancy ranged from 2.2%-87%. The most important risk-factors for alcohol use included tobacco use, partner violence, urban living, and having a male partner who drank alcohol. Only three studies examined changes in alcohol use prior to and following pregnancy recognition with absolute reductions of between 9% and 15%. CONCLUSIONS: Although the burden of alcohol use during pregnancy is likely a significant problem, limited data currently exist for the majority of SSA countries. Furthermore, significant variation likely exists within various populations. Further research is required to explore alcohol use in pregnancy. Strategies to decrease AEP must be developed and implemented in standard pre-natal care.

Genetic diversity in Plasmodium falciparum merozoite surface protein 1 and 2 coding genes and its implications in malaria epidemiology: a review of published studies from 1997-2007.

A major characteristic of human malaria parasites is their genetic diversity and an increasing number of studies have been reported on the epidemiology of Plasmodium falciparum, mainly focusing on the polymorphism of merozoite surface protein (MSP) 1 and 2 genes. A myriad of information on the genetic diversity and multiplicity of P. falciparum infections has been generated from such studies, and a range of molecular tools for epidemiological studies were produced, creating both optimism and pessimism in regard to the global efforts to control malaria. The objective of this review is to provide current and comprehensive information on the diversity in the gene that encodes the merozoite surface protein (MSP) 1 and 2 of P. falciparum and its implications on the epidemiology of malaria, immunity and development of control measures, and point out some research themes that need to be explored further by utilizing molecular techniques currently at our disposal. Articles published in journals between 1997 and 2007 are herein reviewed.

Dynamics of Plasmodium falciparum alleles in children with normal haemoglobin and with sickle cell trait in western Uganda.

We describe the diversity of Plasmodium falciparum populations in western Uganda and assess the role that asymptomatic malaria carriers with sickle cell trait (HbAS) may be playing on the Plasmodium population structure. We genotyped P. falciparum in 291 samples using merozoite surface protein (MSP) 1 and 2 loci. Extensive genetic diversity was detected among symptomatic children in Mbarara (20 MSP1 alleles; 31 MSP2 alleles) and Kagando, Kasese (19 MSP1 alleles; 30 MSP2 alleles). Multiplicity of infection (MOI) was significantly higher in Kagando, Kasese than in Mbarara, with 2.7 and 2.1 genotypes/PCR positive sample with MSP2 marker, respectively. Similar strains were circulating in the two sites; however, a few strains specific to individual sites were observed. Prevalence of HbAS was 36% (12/33) among asymptomatic children in Kisinga sub-county, Kasese. In asymptomatic children, MOI was age-dependent and higher in HbAS carriers than HbAA, suggesting that HbAS carriers harbour a wider range of P. falciparum genotypes. Sickle cell trait may influence rapid acquisition of premunition by creating a reservoir of variant parasite strains in the host. The high level of genetic diversity demonstrated here shows that even in areas with low or seasonal transmission, high levels of parasite polymorphism can occur.