Molecular characterization of HAMP rs10421768 gene and phenotypic expression of hepcidin; a case-control study among sickle cell anaemia patients in Ghana.

BACKGROUND: The sporadic nature of blood transfusion therapy coupled with the alteration of HAMP genes may exacerbate the risk of iron burden in sickle cell anaemia (SCA) patients. The study determined the polymorphic distribution of the HAMP promoter gene rs10421768 and hepcidin levels in SCA patients. METHOD: Sixty participants aged ≥12years [45 SCA patients and 15 controls (HbA)] were recruited from 15th March, 2023 to 20th July, 2023 for a case-control study at Methodist Hospital Wenchi, Ghana. Complete blood count and hepcidin levels assessment were done using haematology analyzer and ELISA, respectively. Genomic DNA was extracted using the Qiagen Kit, and HAMP gene rs10421768 (c.-582 A>G) was sequenced using the MassARRAY method. Data were analysed using SPSS version 26.0. RESULTS: The frequencies of the HAMP promoter rs10421768 genotypes AA, AG, and GG were 64.4%, 33.3%, and 2.2% in SCA patients, and 86.7%, 13.3%, and 0% in the controls, respectively. Serum hepcidin levels were significantly higher among controls than cases [204.0 (154.1-219.3) vs 150.2 (108.1-195.6)µg/L, p<0.010]. Participants with HAMP rs10421768 homozygous A genotype had higher serum levels of hepcidin compared with those in the wild genotypes (AG/GG) group [(188.7 (130.9-226.9) vs 136.8 (109.7-157.8)µg/L, p<0.016]. Disease severity and blood cell parameters were not associated with the HAMP variants (p>0.05). CONCLUSION: The HAMP promoter rs10421768 AA genotype has the highest frequency of distribution and the GG genotype with the least distribution. Participants with HAMP rs10421768 G allele (c.-582A>G) had reduced levels of hepcidin. HAMP rs10421768 genotypes had no association with blood cell parameters and disease severity. The HAMP rs10421768 genotypes may influence serum levels of hepcidin. Further study is required to elucidate the potential effect of the G allele on hepcidin transcription.

Complete Blood Count Reference Intervals for Children Aged Less Than 1 to 12 Years in the Northern Region of Ghana.

Reliable laboratory diagnostic results are key for evaluating and improving children's health. To interpret these results, child-specific reference intervals (RIs), which account for constant biological changes and physiological development with sex and age, are required, as recommended by the Clinical and Laboratory Standards Institute (CLSI). This study presents age- and sex-specific reference intervals for complete blood count (CBC) parameters in children (<1-12 years old) in the Northern Region of Ghana. In this cross-sectional study, 600 healthy children from randomly sampled schools in Tamale (the Northern Region) were recruited and screened. Data from 388 eligible children were used to nonparametrically determine the reference intervals of CBC parameters at the 2.5th and 97.5th percentiles. The CBC reference intervals were compared for variations in sex and age groups using the Wilcoxon rank-sum test. There were no statistically significant differences in most CBC parameters by sex (RBC, Hb, HCT, MCH, RDW (CV/SD), WBC, LYM#, MON#(%) NEU#(%), EOS#(%), and BAS#(%); p > 0.05) and age group (RBC, MCV, RDW (CV/SD), WBC, LYM#, MON#(%) NEU#(%), EOS#(%), and BAS%; p > 0.05). However, there were observable differences between this locally established CBC reference interval and that used for children at Tamale Teaching Hospital (manufacturer's RIs). This study emphasises the importance of determining reference intervals representative of the local child population and incorporating them into the current reporting system of laboratories in the Northern Region to ensure the provision of effective and efficient healthcare services.

Plasma levels of fibrinolytic and coagulation biomarkers in HIV-infected individuals on highly active antiretroviral therapy: A case-control study in a Northern Ghanaian population.

Background and Aim: Impaired coagulation and fibrinolysis have been implicated in thromboembolism in human immunodeficiency virus (HIV)-infected individuals. This study evaluated the plasma levels of plasminogen activator inhibitor-1 (PAI-1) and coagulation biomarkers in HIV-infected individuals on highly active antiretroviral therapy (HAART). Methods: This matched case-control study from March to December, 2020 comprised 76 participants: 38 HIV-positive individuals on HAART and 38 apparently healthy HIV-negative individuals as controls. Blood samples were collected for prothrombin time (PT), activated partial thromboplastin time (aPTT), D-dimers, PAI-1, and soluble fibrin monomer complex (SFMC) estimations. The data were analysed using SPSS version 22.0 and statistical significance was set at p < 0.05. Results: Activated partial thromboplastin time was significantly lower in HIV seropositive individuals on HAART compared with HIV seronegative controls (25.90 s vs. 29.0 s, p = 0.030); however, PT, SFMC, D-dimers, and PAI-1 were significantly higher among the HIV-seropositive individuals compared with the controls: PT: (16.29 s ± 2.16 vs. 15.15 s ± 2.60, p = 0.010), SFMC: [8.53 ng/mL (8.03-9.12) vs. 7.84 ng/mL (7.32-8.58), p = 0.005]), D-Dimer: [463.37 ng/mL (402.70-526.33) vs. 421.11 ng/mL (341.11-462.52), p = 0.015], and PAI-1: [12.77 ng/mL (10.63-14.65) vs. 11.27 ng/mL (10.08-12.95), p = 0.039]. PAI-1 showed a moderate positive correlation with D-Dimer (r = 0.659, p < 0.001) and SFMC (r = 0.463, p = 0.003) among HIV-positive individuals on HAART. There was a strong positive correlation between the plasma PAI-1 concentration and the HIV viral load (r = 0.955, p < 0.001). Conclusion: HIV-seropositive individuals on HAART have deranged coagulation and fibrinolytic markers. Higher HIV viral load correlates strongly with elevated plasma levels of PAI-1 antigens. Periodic assessment of markers of coagulation and fibrinolysis be included in the management of HIV/AIDS in Ghana.

Effects of COVID-19 disease on PAI-1 antigen and haematological parameters during disease management: A prospective cross-sectional study in a regional Hospital in Ghana.

BACKGROUND: Individuals with COVID-19 experience thrombotic events probably due to the associated hypofibrinolysis resulting from the upregulation of plasminogen activator inhibitor-1 (PAI-1) antigen. This study evaluated plasma PAI-1 antigen levels and haematological parameters before treatment and after recovery from severe COVID-19 in Ghana. MATERIALS AND METHODS: This cross-sectional study was conducted at Sunyani Regional Hospital, and recruited 51 patients who had RT-PCR-confirmed SARS-CoV-2. Participants' sociodemographic data and clinical characteristics were taken from the hospital records. Venous blood was taken before COVID-19 treatment commenced for FBC, PAI-1 and ferritin assays. FBC was assessed using an automated haematology analyzer, whilst plasma PAI-1 Ag and serum ferritin levels were assessed with sandwich ELISA. All the tests were repeated immediately after participants recovered from COVID-19. RESULTS: Of the 51 participants recruited into the study, 78.4% (40) had non-severe COVID-19 whiles 21.6% (11) experienced a severe form of the disease. Severe COVID-19 participants had significantly lower haemoglobin (g/dL): 8.1 (7.3-8.4) vs 11.8 (11.0-12.5), p<0.001; RBC x 1012/L: 2.9 (2.6-3.1) vs 3.4 (3.1-4.3), p = 0.001; HCT%: 24.8 ± 2.6 vs 35.3 ± 6.7, p<0.001 and platelet x 109/L: 86.4 (62.2-91.8) vs 165.5 (115.1-210.3), p<0.001, compared with the non-severe COVID-19 group. But WBC x 109/L: 11.6 (9.9-14.2) vs 5.4 (3.7-6.6), p<0.001 and ferritin (ng/mL): 473.1 (428.3-496.0) vs 336.2 (249.9-386.5), p<0.001, were relatively higher in the participants with severe COVID-19 than the non-severe COVID-19 counterparts. Also, the severely ill SARS-CoV-2-infected participants had relatively higher plasma PAI-1 Ag levels (ng/mL): 131.1 (128.7-131.9) vs 101.3 (92.0-116.8), p<0.001, than those with the non-severe form of the disease. Participants had lower haemoglobin (g/dL): 11.4 (8.8-12.3 vs 12.4 (11.5-13.6), p<0.001; RBC x 1012/L: 3.3 (2.9-4.0) vs 4.3 (3.4-4.6), p = 0.001; absolute granulocyte count x 109/L: 2.3 ± 1.0 vs 4.6 ± 1.8, p<0.001, and platelet x 109/L: 135.0 (107.0-193.0) vs 229.0 (166.0-270.0), p<0.001 values at admission before treatment commenced, compared to when they recovered from the disease. Additionally, the median PAI-1 Ag (ng/mL): 89.6 (74.9-100.8) vs 103.1 (93.2-128.7), p<0.001 and ferritin (ng/mL): 242.2 (197.1-302.1) vs 362.3 (273.1-399.9), p<0.001 levels were reduced after a successful recovery from COVID-19 compared to the values at admission. CONCLUSION: Plasma PAI-1 Ag level was higher among severe COVID-19 participants. The COVID-19-associated inflammation could affect red blood cell parameters and platelets. Successful recovery from COVID-19, with reduced inflammatory response as observed in the decline of serum ferritin levels restores the haematological parameters. Plasma levels of PAI-1 should be assessed during the management of severe COVID-19 in Ghana. This will enhance the early detection of probable thrombotic events and prompts Physicians to provide interventions to prevent thrombotic complications associated with COVID-19.

Serum anti-erythropoietin antibodies among pregnant women with Plasmodium falciparum malaria and anaemia: A case-control study in northern Ghana.

BACKGROUND: Anaemia in pregnancy is common in underdeveloped countries, and malaria remains the predominant cause of the condition in Ghana. Anti-erythropoietin (anti-EPO) antibody production may be implicated in the pathogenesis of Plasmodium falciparum malaria-related anaemia in pregnancy. This study ascertained the prevalence of anti-EPO antibody production and evaluated the antibodies' relationship with Plasmodium falciparum malaria and malaria-related anaemia in pregnancy. METHODS: This hospital-based case-control study recruited a total of 85 pregnant women (55 with Plasmodium falciparum malaria and 30 controls without malaria). Venous blood was taken from participants for thick and thin blood films for malaria parasite microscopy. Complete blood count (CBC) analyses were done using an automated haematology analyzer. Sandwich enzyme-linked immunosorbent assay (ELISA) was used to assess serum erythropoietin (EPO) levels and anti-EPO antibodies. Data were analyzed using IBM SPSS version 22.0. RESULTS: Haemoglobin (p<0.001), RBC (p<0.001), HCT (p = 0.006) and platelet (p<0.001) were significantly lower among pregnant women infected with Plasmodium falciparum. Of the 85 participants, five (5.9%) had anti-EPO antibodies in their sera, and the prevalence of anti-EPO antibody production among the Plasmodium falciparum-infected pregnant women was 9.1%. Plasmodium falciparum-infected pregnant women with anti-EPO antibodies had lower Hb (p<0.001), RBC (p<0.001), and HCT (p<0.001), but higher EPO levels (p<0.001). Younger age (p = 0.013) and high parasite density (p = 0.004) were significantly associated with Plasmodium falciparum-related anti-EPO antibodies production in pregnancy. Also, younger age (p = 0.039) and anti-EPO antibody production (p = 0.012) related to the development of Plasmodium falciparum malaria anaemia in pregnancy. CONCLUSION: The prevalence of anti-EPO antibodies among pregnant women with Plasmodium falciparum malaria was high. Plasmodium falciparum parasite density and younger age could stimulate the production of anti-EPO antibodies, and the antibodies may contribute to the development of malarial anaemia in pregnancy. Screening for anti-EPO antibodies should be considered in pregnant women with P. falciparum malaria.

Seroprevalence, trends, and risk factors of hepatitis B and C among family replacement blood donors; a 7-year retrospective study at Sunyani Municipal Hospital, Ghana.

Hepatitis B and C cause chronic infections which develop into liver-related sequelae, like cirrhosis and liver carcinoma. This study determined the seroprevalence, trends, and risk factors of HBV and HCV among family replacement donors. A retrospective review of primary data on blood donors screened between January 2015 and December 2021 was conducted at Sunyani Municipal Hospital. The data were assessed for seroprevalence, trends, and odds ratios using SPSS. Of 6847 donors, the majority were males (88.1% [6033]), ≤24 years (27.4% [1874]), O blood type (69.8% [4776]), and Rh-positive (89.9% [6154]). The seroprevalences of HBV and HCV were 3.2% and 1.9%, respectively, with more males infected with HBV and HCV (3.4% vs 2.0%). Males were 2.842 times (p = .001) and 2.399 times (p = .025) more susceptible than females to HBV and HCV, respectively. In the rainy season, donors were 1.489 times (p = .041) more susceptible to HCV. HBV and HCV seroprevalence declined over the period (slope: -0.5464, p ≤ .001 vs slope: -0.6179, p ≤ .001). Male gender and rainy season were significant determinants of both infections. The seroprevalence of HBV was higher than HCV despite the significant decline in both infections. We, recommend health authorities intensify health education among males and during the rainy season.

Sex-moderated relationship between the 2D:4D ratio and circulating hormones in an adult Ghanaian population.

The second-to-fourth digit ratio (2D:4D) is the putative marker of prenatal hormone exposure. The 2D:4D ratio or the right-left difference (Dr-l) are said to be negative and positive correlates, respectively, of circulating testosterone and estrogen in both adult males and females. However, previous studies on the subject have reported mixed results. This study aimed to determine the sex-moderated relationship between the 2D:4D ratio and adult circulating testosterone, estradiol, testosterone-to-estradiol ratio and the free androgen index. This was a cross-sectional study from January to June 2021 at the University for Development Studies, Ghana. The study involved 62 participants (Female = 28; Male = 34), aged between 20 and 26 years. The right (2D:4DR), the left (2D:4DL), and their difference (Dr-l) were measured by computer-assisted analysis. Fasting venous samples were assayed for total testosterone (T), estradiol (E2 ), and sex hormone-binding globulin (SHBG) using ELISA. The free androgen index (FAI) was then calculated (T/SHBG) and the data were analyzed using moderated and/or weighted regression. Males had significantly higher T and FAI than females while females had significantly higher E2 than males, which were independent of age and body mass index (p < 0.001). There was a significant SEX*Dr-l interaction on FAI (p = 0.007). The Dr-l correlated negatively with FAI in males but positively in females and accounted for about 94.0% of the variability of FAI in males (adjR2  = 0.940) and only 0.2% in females (adjR2  = 0.002). The 2D:4D ratio, a putative marker of prenatal hormone exposure, may have an impact on sex differences in adult free androgen index.

Effect of pulmonary tuberculosis on natural anticoagulant activity in therapy-naïve Ghanaian adults; a case-control study.

Tuberculosis constitutes a global emergency as it affects one-third of the world's inhabitants. Although Pulmonary tuberculosis (PTB) is curable, immunological responses to the infection induce several hematological derangements. This study evaluated the effect of PTB on natural anticoagulant activity and CBC indices. Ninety adults were recruited: 60 PTB patients and 30 non-TB controls. Blood specimens from each participant was tested for Proteins C and S, Antithrombin-III and CBC. Pulmonary TB was associated with significantly reduced Protein C activity (101.46 [87.61-128.3] vs 121.44 [99.50-149.8] IU/L, p= 0.038), RBC (p< 0.0001), HgB (p= 0.0019), HCT (p< 0.0001), MCV (p= 0.0133) and PDW (p< 0.0001) compared to controls. Conversely, PTB patients were associated with significantly increased MCH (p= 0.0086), TWBC (p= 0.0047), Abs. GRAN (p= 0.0226), RDW-CV (p< 0.0001), MCHC (p< 0.0001) and MPV (p= 0.0027) compared to controls. The PTB patients were disproportionately affected with anemia (91.7%, p= 0.001), erythrocytopenia (75.0%, p≤ 0.001) and reduced HCT (80.0%, p≤ 0.001). The frequency of thrombocytosis, leucocytosis, and granulocytosis (50.0%, p= 0.013; 23.3%, p= 0.013; 18.3%, p= 0.025; respectively) in PTB patients were significantly higher than in controls. PTB predisposes to hypercoagulability and causes derangements in erythrocytes, leucocytes, and thrombocytes, and disproportionately causes anemia. Measurement of Protein C activity and CBC indices are useful in the management of PTB patients.

Plasminogen Activator Inhibitor-1 in poorly controlled vs well controlled Type-2 Diabetes Mellitus patients: A case-control study in a district hospital in Ghana.

BACKGROUND: Hypofibrinolysis resulting from the up-regulation of plasminogen activator inhibitor-1 (PAI-1) usually occurs in patients with type 2 diabetes mellitus (T2DM), rendering them hypercoagulable. This study assessed the plasma antigen and activity levels of the PAI-1 enzyme in T2DM patients in a district hospital in Ghana. METHODS: This was a hospital-based case-control study conducted from December 2018 to May 2019 at Nkenkaasu District Hospital. Sixty subjects with T2DM (30 T2DM subjects with good glycemic control and 30 with poor glycemic control), and 30 apparently healthy blood donors were recruited into the study. Blood specimens were collected for complete blood count, lipid profile, PAI-1 Ag and PAI-1 activity levels. A pre-tested questionnaire was used to obtain demographic and clinical information. The data was analyzed using SPSS version 22.0. RESULTS: Elevated PAI-1 Ag and activity levels were observed in the T2DM subjects compared to the healthy controls, with the levels and activity significantly higher (PAI-1 Ag; p< 0.001, PAI-1 activity level; p = 0.004) in the T2DM subjects with poor glycemic control in comparison to those with good glycemic control. A significant positive correlation was observed between HbA1c and PAI-1 enzymes. PAI-1 Ag levels significantly increased along with increased total cholesterol (Β = 0.262, p = 0.033), triglyceride (Β = -0.273, p = 0.034) and HbA1c (Β = 0.419, p = 0.001). Similarly, PAI-1 activity level was associated with total cholesterol (Β = 0.325, p = 0.009), triglyceride (Β = -0.262, p = 0.042), HbA1c (Β = 0.389, p = 0.003) and VLDL-c (Β = -0.227, p = 0.029). CONCLUSION: PAI-1 antigen/activity is enhanced in poorly controlled Ghanaian T2DM subjects. The hypercoagulable state of the affected individuals put them at higher risk of developing cardiovascular diseases. Good glycemic control to regulate plasma PAI-1 levels is essential during T2DM lifelong management. Markers of fibrinolysis should be assessed in these individuals and appropriate anticoagulants given to prevent thrombosis and adverse cardiovascular diseases.

Prevalence and determinants of low testosterone levels in men with type 2 diabetes mellitus; a case-control study in a district hospital in Ghana.

Diabetes mellitus, an endocrine disorder, has been implicated in many including hypogonadism in men. Given the fact that diabetes mellitus is becoming a fast-growing epidemic and the morbidity associated with it is more disabling than the disease itself. This study sought to assess the prevalence of low testosterone levels and predictors in type 2 diabetes mellitus patients and non-diabetic men in a district hospital in Ghana. This hospital-based case-control study comprised 150 type 2 diabetics and 150 healthy men. A pre-structured questionnaire and patient case notes were used to document relevant demographic and clinical information. Venous blood sample of about 6 ml was taken to measure FBS, HbA1c, FSH, LH, and testosterone levels. All data were analyzed using STATA version 12 (STATA Corporation, Texas, USA). The overall hypogonadism in the study population was 48% (144/300). The prevalence of hypogonadism in type 2 diabetic subjects was almost three times more than in healthy men (70.7% vs 25.3%). The odds of having hypogonadism was lower in the men with normal weight and overweight with their underweight counterparts (AOR = 0.33, 95% CI; 0.12-0.96, p = 0.042) and (AOR = 0.29, 95% CI; 0.10-0.84, p = 0.023) respectively. Also, the odds of suffering from hypogonadism was lower in non-smokers compared with smokers (AOR: 0.16, 95% CI; 0.05-0.58, p = 0.005). Participants who were engaged in light (AOR: 0.29, 95% CI; 0.14-0.61, p = 0.001), moderate (AOR: 0.26, 95% CI; 0.13-0.54, p<0.001) and heavy (AOR: 0.25, 95% CI; 0.10-0.67, p = 0.006) leisure time activities had lower odds hypogonadal compared to those engaged in sedentary living. Type 2 diabetic men have high incidence of hypogonadism, irrespective of their baseline clinical, lifestyle or demographic characteristics. Smoking and sedentary lifestyle and BMI were associated with hypogonadism in the study population. Routine testosterone assessment and replacement therapy for high risk patients is recommended to prevent the detrimental effect of hypogonadism in diabetic men.

Molecular Characterization of Glucose-6-Phosphate Dehydrogenase: Do Single Nucleotide Polymorphisms Affect Hematological Parameters in HIV-Positive Patients?

This descriptive, cross-sectional study aimed at evaluating the prevalence of G6PD deficiency and the 376A ⟶ G, 202G ⟶ A single nucleotide polymorphisms (SNPs) among HIV patients attending care at a teaching hospital in Ghana and determine how the SNPs affect haematological profile in HIV. A total of 200 HIV-positive Ghanaians were recruited. Venous blood samples were obtained and complete blood count, and G6PD screening and genotyping for the 376A ⟶ G, 202G ⟶ A SNPs were performed. Out of the 200 participants, 13.0% (26/200) were G6PD-deficient based on the methemoglobin reductase technique, with 1.5% (3/200) and 11.5% (23/200) presenting with partial and full enzyme defect, respectively. Among the 13.0% participants with G6PD deficiency, 19.2% (5/26), 30.8% (8/26), and 19.2% (5/26) presented with 376A ⟶ G only (enzyme activity (EA): 1.19 U/g Hb), 202G ⟶A only (EA: 1.41 U/g Hb), and G202/A376 SNPs (EA: 1.14 U/g Hb), respectively. Having the 376A ⟶ G mutation was associated not only with lower red blood cell (RBC) count (3.38 × 106/µL (3.16-3.46) vs 3.95 × 106/µL (3.53-4.41), p = 0.010) but also with higher mean cell volume (MCV) (102.90 (99.40-113.0) vs 91.10 fL (84.65-98.98), p = 0.041) and mean cell haemoglobin (MCH) (33.70 pg (32.70-38.50) vs 30.75 pg (28.50-33.35), p = 0.038), whereas possessing the 202G ⟶ A mutation was associated with higher MCV only (98.90 fL (90.95-102.35) vs 91.10 fL (84.65-98.98), p = 0.041) compared to G6PD nondeficient participants. The prevalence of G6PD deficiency among HIV patients in Kumasi, Ghana, is 13.0% prevalence, comprising 1.5% and 11.5% partial and full enzyme defect, respectively, based on the methemoglobin reductase technique among HIV patients in Ghana. Among G6PD-deficient HIV patients, the prevalence of G202/A376 SNPs is 19.2%. The 376A ⟶ G mutation is associated not only with lower RBC count but also with higher MCV and MCH, whereas the 202G ⟶ A mutation is associated with higher MCV compared to the normal G6PD population.

Evidence-based region of interest matching guidelines for sarcoma volumetric image-guided radiation therapy.

PURPOSE: Region-of-interest (ROI) guidelines for online cone-beam computed tomography (CBCT) radiotherapy may improve matching reproducibility and reduce inter-user variability of soft tissue sarcoma (STS) image guidance. The purpose of this work is to standardize ROI STS CBCT image registration guidelines using the plan-do-study-act (PDSA) cycle for the lower extremity, retroperitoneal, pelvis, and thorax. METHODS: Based on anatomic bony surrogates, initial ROI matching guidelines for STS were developed by a team of radiation therapists, physicists and oncologists (Plan). Retrospective, qualitative evaluation of the guidelines was completed by the designated sarcoma lead therapist to determine clinical feasibility (Do). Validation of the ROI guidelines was performed through independent evaluation by radiation therapy CBCT imaging experts on a cohort of 10 patients per anatomic region (Study). RESULTS: Draft ROI guidelines were evaluated by 2 independent observers who registered weekly CBCT images to test their validity. Each observer assessed 5 patients per anatomic site, testing ROI options for accuracy of image registration and feasibility, while some ROI borders were adjusted based on algorithm matching performance. Validated ROI guidelines were presented to the sarcoma multidisciplinary site group, and an inter-professional committee of imaging experts for approval prior to clinical implementation (Act). CONCLUSION: ROI matching guidelines for STS IGRT were standardized for 4 anatomic sites using the PDSA cycle for change testing and implementation. IGRT guidelines are intended to improve STS image registration reproducibility, and in turn, are expected to improve the confidence of IGRT decision making and workflow efficiencies for a rare disease with diverse presentation.