Evidence for immune activation in pathogenesis of the HLA class II associated disease, podoconiosis.

Available evidences suggest that podoconiosis is triggered by long term exposure of bare feet to volcanic red clay soil particles. Previous genome-wide studies in Ethiopia showed association between the HLA class II region and disease susceptibility. However, functional relationships between the soil trigger, immunogenetic risk factors and the immunological basis of the disease are uncharted. Therefore, we aimed to characterise the immune profile and gene expression of podoconiosis patients relative to endemic healthy controls. Peripheral blood immunophenotyping of T cells indicated podoconiosis patients had significantly higher CD4 and CD8 T cell surface HLA-DR expression compared to healthy controls while CD62L expression was significantly lower. The levels of the activation markers CD40 and CD86 were significantly higher on monocytes and dendritic cell subsets in patients compared to the controls. RNA sequencing gene expression data indicated higher transcript levels for activation, scavenger receptors, and apoptosis markers while levels were lower for histones, T cell receptors, variable, and constant immunoglobulin chain in podoconiosis patients compared to healthy controls. Our finding provides evidence that podoconiosis is associated with high levels of immune activation and inflammation with over-expression of genes within the pro-inflammatory axis. This offers further support to a working hypothesis of podoconiosis as soil particle-driven, HLA-associated disease of immunopathogenic aetiology.

Common hematological parameters reference intervals for apparently healthy pregnant and non-pregnant women of South Wollo Zone, Amhara Regional State, Northeast Ethiopia.

BACKGROUND: Hematological reference intervals (RIs) are affected by inherent variables like age, sex, genetic background, environment, diet and certain circumstances such as pregnancy signifying the need for population specific values. This study was designed to establish RIs for common hematological parameters of apparently healthy pregnant and non-pregnant women from Northeast Ethiopia. METHOD: This community based cross-sectional study recruited 600 pregnant and non-pregnant women in South Wollo Zone, Northeast Ethiopia from June to August 2019. Complete blood count was performed for eligible participants using Mindary BC-3000 plus hematology analyzer. The mean, median, and 2.5th and 97.5th percentile reference limits with 90% CI were determined using SPSS version 23. RESULT: The established selected 2.5th-97.5th percentiles RIs for pregnant women were: WBC: 4.0-13.2x109/L; RBC: 3.45-4.67x1012/L; Hgb: 10.1-13.7g/dL; HCT: 33.5-46.5%; MCV: 85-104fL; MCH: 27.5-33.0pg; MCHC: 30.3-33.7g/dL and Platelet count: 132-373x109/L. The respective values for non-pregnant women were 3.6-10.3; 4.44-5.01; 12.4-14.3; 38.4-50.1; 86-102; 27.1-32.4, 30.4-34.1, 173-456. A statistically significant difference between pregnant and non-pregnant women was noted in all hematological parameters except MCHC. The mean and median value of WBC count, MCV, MPV and PDW increased, whereas mean values of HCT and Platelet count decreased as gestational period advances. CONCLUSION: The observed difference from other studies signify the necessity for using trimester specific RIs and separate RI for pregnant and non-pregnant women. Thus, we recommend the health facilities in the study area to utilize the currently established RIs for pregnant and non-pregnant women for better care.

Effect of Gasoline Exposure on Hematological Parameters of Gas Station Workers in Mekelle City, Tigray Region, Northern Ethiopia.

BACKGROUND: The adverse health effects of chronic gasoline exposure may be related to impairment of the hematopoietic system with bone marrow suppression, an increased risk of blood cell morphology abnormality and developing cancer. OBJECTIVE: To assess the effect of gasoline exposure on hematological parameters among gas station workers in Mekelle City, Tigray Region, Northern Ethiopia. METHODS: This cross-sectional study was carried out on 43 subjects (exposed group) and 77 subjects (unexposed group) with matched age and sex. Socio-demographic characteristics and duration of exposure data were collected using a structured questionnaire and an observation checklist. Sysmex XP-300 was used for hematological analysis and stained peripheral blood smear was examined for any abnormality. Data were entered and analyzed using SPSS version 23. RESULTS: Of exposed individuals, 28/43 (65.1%) and 49/77 (63.6%) of controls were males. The average exposure time was 5.19±4.38 years, with an average working hour of 11.74±1.89 hours/day. The mean RBC count (1012/L), HCT (%), HGB (g/dl) and platelets count (109/L) of the exposed group were significantly lower (4.88±0.573, 43.29±3.71, 15.04±1.33 and 248.95±58.19) compared with controls (5.35±0.533, 44.95±3.10, 15.59±1.26 and 292.45±62.17) at p<0.05, respectively. The MCH (pg) (30.48±2.06 vs 29.52±1.66) and MCHC (g/dl) (34.83±0.988 vs 34.32±0.927) were significantly higher in the exposed group compared with controls (p<0.05). HCT, RBC, HGB and platelet counts were significantly decreased with increased years of exposure (p<0.05). The peripheral blood film examination revealed basophilic stippling and macrocytosis in 9.3% of the exposed group. CONCLUSION: Long-term exposure to gasoline at gas stations affected RBC parameters and platelet count. A significant negative correlation was noted between duration of exposure and HGB, HCT and platelet count, warranting implementation of protective measures at gas stations.

Rapid ethical appraisal of stakeholder views on research prior to undertaking immunopathogenesis studies on podoconiosis in northeast Ethiopia during a period of social instability.

BACKGROUND: Undertaking research and attaining informed consent can be challenging when there is political unrest and community mistrust. Rapid ethical appraisal (REA) is a tool that uses qualitative methods to explore sociocultural issues that may affect the ethical conduct of research. METHODS: We used REA in northeast Ethiopia shortly following a period of unrest, during which violence against researchers occurred, to assess stakeholder perceptions of research, researchers and the informed consent process. We held 32 in-depth interviews and 2 focus group discussions. RESULTS: Most community members had little awareness about podoconiosis or healthcare research. Convincing the community to donate blood for research is challenging due to association with HIV testing. The attack on researchers was mainly motivated by the community's mistrust of their intentions against the background of a volatile political situation. Social media contributed to the spread of misinformation. Lack of community engagement was also a key contributing factor. CONCLUSIONS: Using REA, we identified potential barriers to the informed consent process, participant recruitment for data and specimen collection and the smooth conduct of research. Researchers should assess existing conditions in the study area and engage with the community to increase awareness prior to commencing their research activities.

Role of platelet parameters in early detection and prediction of severity of preeclampsia: A comparative cross-sectional study at Ayder comprehensive specialized and Mekelle general hospitals, Mekelle, Tigray, Ethiopia.

BACKGROUND: Platelet parameters alterations are one of the most commonly identified hematological changes in preeclampsia (PE). However, their functions as a tool for prediction and prognosis of PE have not been extensively studied in developing countries. The aim of this study was to compare platelet count (PC), and platelet indices (mean platelet volume (MPV), platelet distribution width (PDW), and platelet large cell ratio (PLCR)) between preeclamptic and normotensive (NT) pregnant women and assess their role in diagnosis and prediction of PE development. METHODS: A cross sectional comparative study was conducted at Ayder comprehensive specialized hospital (ACSH) and Mekelle general hospital (MGH) from January to March 2017. Platelet parameters of mild preeclamptic (mPE) (n = 35), severe preeclamptic (sPE) (n = 44) and NT pregnant women (n = 140) were analyzed using SYSMEX-XT 4000i automated hematology analyzer. One-way ANOVA supplemented with post-hoc test, receiver operating characteristics (ROC) curve and pearson correlation test statistical analyses were performed. P < 0.05 was considered significant. RESULT: Pregnant women with sPE had lower PC as compared with that of mPE and NT women (p<0.05). All platelet indices showed significant increment with severity of PE. PC was negatively correlated with platelet indices. There was a positive correlation among platelet indices. ROC analysis revealed that MPV had the largest area under the ROC curve (0.85; 95%CI (0.79, 0.89)) with cutoff value >9.45fl, sensitivity of 83.5%, specificity of 86.4%, positive predictive value of 77.6% and negative predictive value of 90.3%. CONCLUSION: MPV and PC were identified as good candidates for sPE diagnosis. Because evaluation of platelet parameters is rapid, reliable and economical, they can be utilized as an alternative biomarker for prediction and prognosis of PE.

Effect of iron deficiency anemia on HbA1c in diabetic patients at Tikur Anbessa specialized teaching hospital, Addis Ababa Ethiopia.

BACKGROUND: Hemoglobin A1C (HbA1c) is the predominant hemoglobin found in HbA1 fractions. A1c assay is the recommended assay for diagnosing diabetes and any condition that changes red cell turnover such as Iron deficiency Anemia (IDA), will lead to spurious A1C results. Therefore, the present study was aimed at determining the effect of IDA on HbA1c in diabetic patients attending Black Lion Specialized Teaching Hospital, Addis Ababa, Ethiopia. METHODS: A facility based comparative cross sectional study was conducted on 174 diabetic patients (87 with IDA and 87 without IDA) from April to July 2016. Socio demographic data and clinical conditions were collected using structured questionnaire. Venous blood was collected for performing Complete blood count (CBC) using Cell dyn 1800 hematology analyzer; Serum ferritin, performed by COBAS INTEGRA 400/800 Chemistry analyzer and HbA1c tests, performed by COBAS C 111 analyzer. Data was analyzed using SPSS version 21 software. Pearson's correlation, chi-square, and independent t-tests were calculated. The data was presented as mean ± SD. A P-value of < 0.05 was taken as statistically significant. RESULTS: Mean hemoglobin (Hgb), hematocrit (HCT), Mean cell volume (MCV), mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC) were lower in IDA group compared to non-IDA diabetic patients. HbA1c (%) level was significantly lower in IDA group (6.18 ± 1.57) compared with the non-IDA diabetic patients (7.74 ± 1.81) (p < 0.05). CONCLUSION: HbA1c is significantly lower in diabetic patients with IDA compared to the non-IDA diabetic patients. Therefore, the authors believe that monitoring these patients using only HbA1c could be misleading, hence physicians and health care providers should take this into account before making any therapeutic decision. Detailed examination including large number of participants employing advanced laboratory techniques is recommended.

Phenotypic and functional heterogeneity of peripheral γδ T cells in pulmonary TB and HIV patients in Addis Ababa, Ethiopia.

BACKGROUND: Previous studies reported HIV infection alters the distribution and function of γδ T cells and their subsets. γδ T phenotypes in healthy and diseased individuals has received little attention in Ethiopia. We conducted this study to analyze the distribution of γδ T cells, the subsets and levels of expression of activation (CD38), exhaustion or anergy (CD95, PD1), adhesion (N-CAM/CD56 and CD103), among HIV and TB infected patients. METHOD: The distributions of total γδ T cells, Vδ1 and Vδ2 T cells subsets were analyzed in clinical samples collected from asymptomatic HIV, pulmonary TB patients and apparently healthy controls. Multicolor flow cytometry and IFN-γ ELISA were used to assess surface markers and functional responses of Vδ2 T cells to isopentenyl pyrophosphate stimulation, respectively. RESULT: A total of 52 study participants were enrolled in this study, 22 HIV + TB-, 10 HIV-TB+ and 20 healthy controls. No significant differences were observed in the distribution of total γδ T cells and in the proportion of Vδ1 subsets in all study groups, though slightly higher proportions were observed in HIV + TB- patients for the latter, of borderline statistical significance (p = 0.07). However, the proportion of Vδ2 T cells, as well as the IFN-γ response to IPP stimulation, was significantly reduced in HIV + TB- patients compared to healthy controls (p < 0.002). Expression of the activation marker CD38 (p < 0.001) and adhesion marker CD103 (αEß7) were significantly higher in the Vδ1 T cell subset among both HIV + TB- (p = 0.013) and HIV-TB+ (p = 0.006) patients compared to healthy controls. Similarly, exhaustion markers, CD95 and PD1, were significantly higher in these two T cell subsets among both HIV + TB- and HIV-TB+ patients (p < 0.01). Interestingly, we also observed an increased proportion of effector memory (CD45RA-CD27-) and effector cytotoxic (CD45RA + CD27-) Vδ2 T cell subsets in HIV negative pulmonary TB patients. CONCLUSION: In sum, HIV infection was associated with an increase in Vδ1 and a decrease in the function and frequencies of Vδ2 T cells. Moreover, increased effector Vδ2 T cells were observed among HIV negative pulmonary TB patients suggesting a potential role of these T cells in the host response to TB.

Diagnostic predictive value of platelet indices for discriminating hypo productive versus immune thrombocytopenia purpura in patients attending a tertiary care teaching hospital in Addis Ababa, Ethiopia.

BACKGROUND: Bone marrow examination may be required to discriminate causes of thrombocytopenia as hypoproductive or hyperdestructive. However, this procedure is invasive and time consuming. This study assessed the diagnostic value of Mean Platelet Volume (MPV), Platelet Distribution Width (PDW) and Platelet Large Cell-Ratio (P-LCR) in discriminating causes of thrombocytopenia as hypoproductive or hyperdestructive (Immune thrombocytopenia purpura). METHOD: A prospective cross-sectional study was conducted on 83 thrombocytopenic patients (Plt < 150 × 10(9)/L). From these, 50 patients had hypoproductive and the rest 33 Immune Thrombocytopenia Purpura (ITP). Age and sex matched 42 healthy controls were included as a comparative group. Hematological analysis was carried out using Sysmex XT 2000i 5 part diff analyzer. SPSS Version16 was used for data analysis. A two by two table and receiver operating characteristic (ROC) curve was used to calculate sensitivity, specificity, positive and negative predictive values, for a given platelet indices (MPV, PDW and P-LCR). Student t test and Mann Whitney U test were used to compare means and medians, respectively. Correlation test was used to determine associations between continuous variables. RESULTS: All Platelet indices were significantly higher in ITP patients (n = 33) than in hypoproductive thrombocytopenic patients (n = 50) (p < 0.0001). In particular MPV and P-LCR have larger area under ROC curve (0.876 and 0.816, respectively), indicating a better predictive capacity, sensitivity and specificity in discriminating the two causes of thrombocytopenia. The indices were still significantly higher in ITP patients compared to 42 healthy controls (p < 0.0001). A significant negative correlation was observed between platelet count and platelet indices in ITP patients, (p < 0.001). CONCLUSION: MPV, PDW and P-LCR help in predicting thrombocytopenic patients as having ITP or hypoproductive thrombocytopenia. If these indices are used in line with other laboratory and clinical information, they may help in delaying/ avoiding unnecessary bone marrow aspiration in ITP patients or supplement a request for bone morrow aspiration or biopsy in hypoproductive thrombocytopenic patients.