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.

Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity.

BACKGROUND: Plasmodium falciparum, the parasite causing malaria, affects populations in many endemic countries threatening mainly individuals with low malaria immunity, especially children. Despite the approval of the first malaria vaccine Mosquirix™ and very promising data using cryopreserved P. falciparum sporozoites (PfSPZ), further research is needed to elucidate the mechanisms of humoral immunity for the development of next-generation vaccines and alternative malaria therapies including antibody therapy. A high prevalence of antibodies against AMA1 in immune individuals has made this antigen one of the major blood-stage vaccine candidates. MATERIAL AND METHODS: Using antibody phage display, an AMA1-specific growth inhibitory human monoclonal antibody from a malaria-immune Fab library using a set of three AMA1 diversity covering variants (DiCo 1-3), which represents a wide range of AMA1 antigen sequences, was selected. The functionality of the selected clone was tested in vitro using a growth inhibition assay with P. falciparum strain 3D7. To potentially improve affinity and functional activity of the isolated antibody, a phage display mediated light chain shuffling was employed. The parental light chain was replaced with a light chain repertoire derived from the same population of human V genes, these selected antibodies were tested in binding tests and in functionality assays. RESULTS: The selected parental antibody achieved a 50% effective concentration (EC50) of 1.25 mg/mL. The subsequent light chain shuffling led to the generation of four derivatives of the parental clone with higher expression levels, similar or increased affinity and improved EC50 against 3D7 of 0.29 mg/mL. Pairwise epitope mapping gave evidence for binding to AMA1 domain II without competing with RON2. CONCLUSION: We have thus shown that a compact immune human phage display library is sufficient for the isolation of potent inhibitory monoclonal antibodies and that minor sequence mutations dramatically increase expression levels in Nicotiana benthamiana. Interestingly, the antibody blocks parasite inhibition independently of binding to RON2, thus having a yet undescribed mode of action.

Coagulation abnormalities in childhood acute lymphoblastic leukemia: assessing the impact of L-asparaginase therapy in Ghana.

BACKGROUND: Although the rate of childhood acute lymphoblastic leukemia (ALL) is increasing in Africa, there is a dearth of information on the disease and the dynamics of hemostatic parameters with therapy. METHODS: In this case-control study, we evaluated variations in the level/activity of selected coagulation parameters among cALL in Ghana and healthy controls stratified by stage of therapeutic management. RESULTS: In all, the research recruited 104 participants comprising 26 cALL cases and 78 healthy controls. The cALL group had significantly higher prothrombin time (PT) (p = 0.001), activated partial thromboplastin time (APTT) (p < 0.0001) and D-dimers (p = 0.001) but lower platelet (PLT) count, protein C (PC) (p < 0.0001), protein S (PS) (p < 0.0001) and antithrombin III (ATIII) (p < 0.0001) compared to controls. Compared to the healthy controls, activity of PC was lower during induction (p < 0.0001), consolidation (p = 0.005) and maintenance phases of chemotherapy (p = 0.012) while activities of PS and ATIII were lower at both induction (p < 0.0001, p = 0.006) and consolidation (p < 0.0001, p = 0.018) phases of chemotherapy. CONCLUSION: Our findings provide evidence in the context of Africa and corroborates previous reports that cALL could result in a state of hypercoagulability, possibly leading to a high risk of thrombosis and thromboembolic complications. This possibly increased risk is not limited to the induction phase but also the consolidation phase.

Assessment of platelet indices and platelet activation markers in children with Plasmodium falciparum malaria.

BACKGROUND: Plasmodium falciparum malaria remains one of the world's major infectious diseases that cause most morbidity and mortality, particularly in children. In Ghana, most children below the ages of 5 years depending on the severity of the infection often lose their lives. However, it is still debatable why infection with falciparum malaria contributes to thrombocytopenia. METHODS: This study sought to investigate the expression of the various platelet indices and activation markers in children with falciparum malaria. Platelet indices (Platelet count [PLT], Plateletcrite [PCT], Mean Platelet Volume [MPV], Platelet Distribution Width [PDW] and Platelet-Large Cell Ratio [P-LCR]) and platelet surface membrane glycoproteins (GPIIb/IIIa [PAC-1], P-selectin [CD62p] and GPIV [CD36]) expressions were determined in children with falciparum malaria (cases) and healthy children (controls) using automated blood cell analysis and flow cytometry techniques, respectively. RESULTS: Except for P-LCR, all the other platelet indices (PLT, MPV, PDW, and PCT) were significantly lower in the cases than the controls (P < 0.05). Also, it was observed that the level of expression of the activation markers; PAC 1 and CD62p showed a significant (P < 0.05) decreased before and after activation in falciparum malaria cases than in the controls. On the contrary, CD36 expression in the controls did not differ significantly (p > 0.05) from the malaria cases. Platelet activation markers were known to be associated with increased risk of falciparum malaria with the mean fluorescence intensity of PAC1 (Odds Ratio [OR] 34.0, Relative Risk [RR] 4.47, 95% Confidence Interval [CI] 4.904-235.7; p < 0.0001) and CD36 (OR 4.2, RR 1.82, 95% CI 0.9824-17.96; p = 0.04). Moreover, the percentage expression of CD62p (OR 4.0, RR 1.80, 95% CI 0.59-27.24; p = 0.19) was also observed to be probably associated with increased risk of falciparum malaria although not statistically significant (p > 0.05). CONCLUSION: Plasmodium falciparum malaria has been known to be associated with platelet activation markers, which probably contributes to thrombocytopenia.

Plasmodium falciparum histidine-rich protein 2 diversity in Ghana.

BACKGROUND: In the absence of microscopy, Plasmodium falciparum histidine-rich proteins 2 (PfHRP2)-based rapid diagnostic tests (RDTs) are recommended for the diagnosis of falciparum malaria, particularly in endemic regions. However, genetic variability of the pfhrp2 gene threatens the usefulness of the test due to its impact on RDT sensitivity. This study aimed to investigate the diversity of pfhrp2 in malaria cases among children in Ghana. METHODS: A cross-sectional study was conducted at the Adidome Government Hospital in the Volta Region of Ghana. A total of 50 children with mean age of 6.6 ± 3.5 years and diagnosed falciparum malaria were included. Blood samples were collected for complete blood count, malaria parasite identification and counting using auto analyzer and microscopy, respectively. DNA was isolated from blood-spotted Whatman filters, amplified and sequenced. Nucleotide sequences were translated in silico to corresponding amino acids and the deduced amino acids sequences were analyzed for diversity using Mega X. RESULTS: The number of repeats and number of each repeat within PfHRP2 varied between isolates. Twelve rare PfHRP2 repeat types, two of which are previously unreported, were identified in this study. The HRP2 sequence obtained in this study shared high similarities with isolates from Kenya. Using Baker's regression model, Group B was the highest occurring type (58.0%). Screening of all sequences for epitopes recognized by PfHRP2-specific monoclonal antibodies (mAbs), the predominant motif was AHHAADAHH, which is recognized by the C1-13 mAbs. CONCLUSION: This study reports diversity of P. falciparum HRP2 in samples from Ghanaian children with symptomatic malaria. The findings of this study highlight the existence of extra amino acid repeat types which adds to the PfHRP2 antigenic variability.

Antierythropoietin Antibody Production Is Not Associated with Malaria and Malaria-Related Anaemia in Humans.

INTRODUCTION: The pathophysiology of malaria-related anaemia is not fully understood although increased destruction of parasitized and nonparasitized erythrocytes, as well as inadequate erythropoiesis, has been proposed. Circulating antierythropoietin (anti-EPO) antibodies have also been implicated in malaria and malaria-related anaemia in mice. However, studies on this association have not been investigated in humans. This study therefore determined the prevalence of anti-EPO antibody production and assessed its association with malaria and malaria-related anaemia in humans. METHODS: A total of 86 children aged 1-10 years (57 children with malaria serving as the case group and 29 healthy children serving as control), all residents of Duayaw Nkwanta, Ghana, were recruited for this case-control study. Venous blood was collected for thick and thin films for malaria microscopy, full blood count by automated haematology analyzer, and antierythropoietin antibody and erythropoietin estimation by sandwich ELISA method. RESULTS: Out of the 86 participants recruited, only 3 (3.5%) were positive for anti-EPO antibody; 2.3% of the case group; and 1.2% of the control group. There was no association between the cases and the controls in the production of anti-EPO antibodies. Erythropoietin concentration was significantly higher in malaria-related anaemic subjects (p=0.032). CONCLUSION: Antierythropoietin antibodies are not associated with malaria infection and malaria-related anaemia in humans. Erythropoietin concentration is associated with malaria-related anaemia.

Predominant Complications of Type 2 Diabetes in Kumasi: A 4-Year Retrospective Cross-Sectional Study at a Teaching Hospital in Ghana.

Background and objectives: Diabetes mellitus type 2 (T2DM) has been associated with several microvascular and macrovascular complications. However, studies regarding the predominant complications of T2DM in Ghana have not been conducted. This study evaluated the prevalence and predominant complications of T2DM and assessed the sociodemographic factors associated with the development of diabetes-related complications in Kumasi, Ghana. Materials and Methods: This was a retrospective cross-sectional study conducted at Komfo Anokye Teaching Hospital (KATH), Kumasi, Ghana. A total of 1600 Ghanaian T2DM adults were included in this study. Patients' clinical data from 2012 to 2016 were retrieved from the hospital's archive. Results: The prevalence of macrovascular and microvascular complications of T2DM was 31.8% and 35.3% respectively. The prevalence of neuropathy, nephropathy, retinopathy, sexual dysfunction, diabetic keto-acidosis (DKA), and hypoglycemia were 20.8%, 12.5%, 6.5%, 3.8%, 2.0%, and 0.8% respectively. Sexual dysfunction was significantly associated with the male gender compared to females. Being employed: Informal (aOR = 0.479, p < 0.0001), and Formal (aOR = 0.475, p = 0.0008) was associated with lower age- and sex-adjusted odds of developing T2DM-related complications while having T2DM for 5-10 years (aOR = 1.550, p = 0.0009) and more than 10 years (aOR = 2.755, p < 0.0001) was associated with increased odds of developing complications. Conclusions: Microvascular complication is the most predominant among T2DM in Kumasi, Ghana. The most prevalent T2DM-related microvascular complication in Kumasi, Ghana is neuropathy. Sexual dysfunction is associated with male compared to female T2DM patients. Being employed reduces the chance of developing T2DM-related complications while increasing DM duration increases the risk of complications.

Acquired immune responses to three malaria vaccine candidates and their relationship to invasion inhibition in two populations naturally exposed to malaria.

BACKGROUND: Malaria still represents a major cause of morbidity and mortality predominantly in several developing countries, and remains a priority in many public health programmes. Despite the enormous gains made in control and prevention the development of an effective vaccine represents a persisting challenge. Although several parasite antigens including pre-erythrocytic antigens and blood stage antigens have been thoroughly investigated, the identification of solid immune correlates of protection against infection by Plasmodium falciparum or clinical malaria remains a major hurdle. In this study, an immuno-epidemiological survey was carried out between two populations naturally exposed to P. falciparum malaria to determine the immune correlates of protection. METHODS: Plasma samples of immune adults from two countries (Ghana and Madagascar) were tested for their reactivity against the merozoite surface proteins MSP1-19, MSP3 and AMA1 by ELISA. The antigens had been selected on the basis of cumulative evidence of their role in anti-malarial immunity. Additionally, reactivity against crude P. falciparum lysate was investigated. Purified IgG from these samples were furthermore tested in an invasion inhibition assay for their antiparasitic activity. RESULTS: Significant intra- and inter- population variation of the reactivity of the samples to the tested antigens were found, as well as a significant positive correlation between MSP1-19 reactivity and invasion inhibition (p < 0.05). Interestingly, male donors showed a significantly higher antibody response to all tested antigens than their female counterparts. In vitro invasion inhibition assays comparing the purified antibodies from the donors from Ghana and Madagascar did not show any statistically significant difference. Although in vitro invasion inhibition increased with breadth of antibody response, the increase was not statistically significant. CONCLUSIONS: The findings support the fact that the development of semi-immunity to malaria is probably contingent on the development of antibodies to not only one, but a range of antigens and that invasion inhibition in immune adults may be a function of antibodies to various antigens. This supports strategies of vaccination including multicomponent vaccines as well as passive vaccination strategies with antibody cocktails.

Isolation, production and characterization of fully human monoclonal antibodies directed to Plasmodium falciparum MSP10.

BACKGROUND: Semi-immunity against the malaria parasite is defined by a protection against clinical episodes of malaria and is partially mediated by a repertoire of inhibitory antibodies directed against the blood stage of Plasmodium falciparum, in particular against surface proteins of merozoites, the invasive form of the parasite. Such antibodies may be used for preventive or therapeutic treatment of P. falciparum malaria. Here, the isolation and characterization of novel human monoclonal antibodies (humAbs) for such applications is described. METHODS: B lymphocytes had been selected by flow cytometry for specificity against merozoite surface proteins, including the merozoite surface protein 10 (MSP10). After Epstein-Barr virus (EBV) transformation and identification of promising resulting lymphoblastoid cell lines (LCLs), human immunoglobulin heavy and light chain variable regions (Vh or Vl regions) were secured, cloned into plant expression vectors and transiently produced in Nicotiana benthamiana in the context of human full-size IgG1:κ. The specificity and the affinity of the generated antibodies were assessed by ELISA, dotblot and surface plasmon resonance (SPR) spectroscopy. The growth inhibitory activity was evaluated based on growth inhibition assays (GIAs) using the parasite strain 3D7A. RESULTS: Supernatants from two LCLs, 5E8 and 5F6, showed reactivity against the second (5E8) or first (5F6) epidermal growth factor (EGF)-like domain of MSP10. The isolated V regions were recombinantly expressed in their natural pairing as well as in combination with each other. The resulting recombinant humAbs showed affinities of 9.27 × 10(-7) M [humAb10.1 (H5F6:κ5E8)], 5.46 × 10(-9) M [humAb10.2 (H5F6:κ5F6)] and 4.34 × 10(-9) M [humAb10.3 (H5E8:κ5E8)]. In GIAs, these antibodies exhibited EC50 values of 4.1 mg/ml [95% confidence interval (CI) 2.6-6.6 mg/ml], 6.9 mg/ml (CI 5.5-8.6 mg/ml) and 9.5 mg/ml (CI 5.5-16.4 mg/ml), respectively. CONCLUSION: This report describes a platform for the isolation of human antibodies from semi-immune blood donors by EBV transformation and their subsequent characterization after transient expression in plants. To our knowledge, the presented antibodies are the first humAbs directed against P. falciparum MSP10 to be described. They recognize the EGF-like folds of MSP10 and bind these with high affinity. Moreover, these antibodies inhibit P. falciparum 3D7A growth in vitro.

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.

Association between micronutrients, oxidative stress biomarkers and angiogenic growth mediators in early and late-onset preeclamptic Ghanaian women.

Objectives: Micronutrients, especially calcium (Ca) and magnesium (Mg) are reported to reduce preeclampsia events via several factors such as endothelial cell control, optimal oxidative stress and a balanced angiogenic growth mediator. We evaluated the association of micronutrients with oxidative stress biomarkers, and angiogenic growth mediators in early-onset preeclampsia and late-onset preeclampsia. Methods: This case-control study recruited 197 preeclampsia (early-onset preeclampsia = 70 and late-onset preeclampsia = 127) as cases and 301 normotensive pregnant women as controls from the Komfo Anokye Teaching Hospital, Ghana. Samples were collected after 20 weeks of gestation for both cases and controls and estimated for Ca, Mg, soluble fms-like tyrosine kinase-1, placental growth factor, vascular endothelial growth factor-A, soluble endoglin, 8-hydroxydeoxyguanosine, 8-epiprostaglandinF2-alpha and total antioxidant capacity. Results: Early-onset preeclampsia women had significantly lower levels of Ca, Mg, placental growth factor, vascular endothelial growth factor-A and total antioxidant capacity but higher levels of soluble fms-like tyrosine kinase-1, soluble endoglin, 8-epiprostaglandinF2-alpha, 8-hydroxydeoxyguanosine, soluble fms-like tyrosine kinase-1/placental growth factor ratio, 8-epiprostaglandinF2-alpha /placental growth factor ratio, 8-hydroxydeoxyguanosine/placental growth factor ratio and soluble endoglin/placental growth factor ratio than late-onset preeclampsia and normotensive pregnant women (p < 0.0001). Among the early-onset preeclampsia women, the first and second quartile for serum placental growth factor, first quartile for vascular endothelial growth factor-A and total antioxidant capacity and the fourth quartiles for serum sEng, serum sFlt-1, 8-epiPGF2α and 8-OHdG were independently associated with low Ca and Mg (p < 0.05). Among late-onset preeclampsia women, the fourth quartile for soluble fms-like tyrosine kinase-1 was independently associated with low Ca and Mg (p < 0.05). Conclusion: Magnesium and calcium are associated with an imbalance in angiogenic growth mediators and oxidative stress biomarkers among preeclampsia women, particularly early-onset preeclampsia. Serial and routine measurement of these micronutrients would allow the monitoring of poor placental angiogenesis while enabling an understanding of the triggers of increased oxidative stress and reduced antioxidant in preeclampsia.

Biomarkers of oxidative stress and its nexus with haemoglobin variants and adverse foeto-maternal outcome among women with preeclampsia in a Ghanaian population: A multi-centre prospective study.

INTRODUCTION: Haemoglobin variants and preeclampsia (PE) are associated with adverse fatal events of which oxidative stress may be an underlying factor. Oxidative stress (OS) among preeclamptic women with haemoglobin variants has been well established. It is, however, unclear whether haemoglobin variants induce OS to aggravate the risk of adverse foeto-maternal outcomes in pregnant women with preeclampsia. We measured the levels of OS biomarkers and determined the association between haemoglobin variants, and adverse foeto-maternal outcomes among pregnant women with PE. METHODS: This multi-centre prospective study recruited 150 PE women from three major health facilities in both Bono and Bono east regions of Ghana from April to December 2019. Haemoglobin variants; HbAS, HbSS, HbSC, HbCC, and HbAC were determined by haemoglobin electrophoresis. OS biomarkers such as malondialdehyde (MDA), catalase (CAT), vitamin C, and uric acid (UA) along with haematological and biochemical parameters were estimated using standard protocol. Adverse pregnancy complications (APCs) such as post-partum haemorrhage (PPH), HELLP (Haemolysis, Elevated liver enzymes, Low platelet count) syndrome, preterm delivery, neonatal intensive care unit (NICU) admission, and neonatal jaundice were recorded. RESULTS: Of the 150 pregnant women with preeclampsia, the distribution of haemoglobin AA, AS, AC, CC, SS and SC phenotypes were 66.0%, 13.3%, 12.7%, 3.3%, 3.3% and 1.3%, respectively. The most prevalent foeto-maternal outcomes among PE women were NICU admission (32.0%) followed by PPH (24.0%), preterm delivery (21.3%), HELLP syndrome (18.7%), and neonatal jaundice (18.0%). Except for vitamin C level which was significantly higher in patients with at least a copy of Haemoglobin S variant than those with at least a copy of Haemoglobin C variant (5.52 vs 4.55; p = 0.014), levels of MDA, CAT, and UA were not statistically significantly different across the various haemoglobin variants. Multivariate logistic regression model showed that participants with HbAS, HbAC, having at least a copy of S or C and participants with HbCC, SC, SS had significantly higher odds of neonatal jaundice, NICU admission, PPH and HELLP syndrome compared to participants with HbAA. CONCLUSION: Reduced levels of vitamin C are common among preeclamptics with at least one copy of the HbC variant. Haemoglobin variants in preeclampsia contribute to adverse foeto-maternal outcomes with Haemoglobin S variants being the most influencing factor for PPH, HELLP, preterm labour, NICU admission, and neonatal jaundice.

Quality of glycemic control in type 2 diabetes mellitus (T2DM) and its association with markers of coagulation and inhibitors of fibrinolysis: A case-control study in the Upper West Region, Ghana.

Background and Aims: Type 2 diabetes mellitus (T2DM) individuals are at a higher risk of developing diabetes complications, with approximately 80% complication-related mortality. The increased morbidity and mortality among T2DM patients are partly due to dysregulated hemostasis. This study determined the quality of glycemic control in T2DM and its association with markers of coagulation and inhibitors of fibrinolysis. Methods: This case-control study recruited 90 participants involving: 30 T2DM patients with good glycemic control, 30 with poor glycemic control, and 30 nondiabetic subjects as controls at a Municipal Hospital in Ghana. Fasting blood glucose, glycated hemoglobin, activated partial thromboplastin time (APTT), prothrombin time (PT), calculated international normalized ratio (INR), and full blood count (FBC) were determined for each respondent. Plasma levels of plasminogen activator inhibitor-1 (PAI-1) and thrombin activatable fibrinolysis inhibitor (TAFI) were determined using the solid-phase sandwich enzyme-linked immunosorbent assay method. Data were analyzed using R language software. Results: Plasma PAI-1 antigen levels were significantly higher in the participants with poor glycemic control as compared to participants with good glycemic control (p < 0.0001). There was no significant difference in plasma TAFI levels between the participants with poor glycemic control as compared to participants with good glycemic control (p = 0.900). T2DM patients had significantly shorter APTT, PT, and INR than controls (p < 0.05). At a cut-off of ≥161.70 pg/µL, PAI was independently associated with increasing odds (adjusted odds ratio = 13.71, 95% confidence interval: 3.67-51.26, p < 0.0001) of poor glycemic control and showed the best diagnostic accuracy for poor glycemic control (area under the curve = 0.85, p < 0.0001). Conclusion: PAI-1 levels were significantly increased in T2DM with poor glycemic control and emerged as the best predictor for poor glycemic control. Good glycemic management to control the plasma levels of PAI-1 is required to prevent hypercoagulability and thrombotic disorders.

Coagulation Factors and Natural Anticoagulants as Surrogate Markers of Preeclampsia and Its Subtypes: A Case-Control Study in a Ghanaian Population.

Preeclampsia (PE) is associated with endothelial injury and hemostatic abnormalities. However, the diagnostic role of coagulation parameters and natural anticoagulants in predicting PE has not been explored in Ghana. This study assessed plasma levels of these factors as surrogate markers of PE and its subtypes. This case-control study included 90 women with PE (cases) and 90 normotensive pregnant women (controls). Blood samples were drawn for the estimation of complete blood count and coagulation tests. The prothrombin time (PT), activated partial thromboplastin time (APTT), and the calculation of the international normalized ratio (INR) were determined by an ACL elite coagulometer while the levels of protein C (PC), protein S (PS), antithrombin III (ATIII), and D-dimers were also measured using the solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) method. All statistical analyses were performed using the R Language for Statistical Computing. Results showed significantly (p < .05) shortened APTT (28.25 s) and higher D-dimer levels (1219.00 ng/mL) among PE women, as well as low levels of PC (1.02 µg/mL), PS (6.58 µg/mL), and ATIII (3.99 ng/mL). No significant difference was found in terms of PT and INR. From the receiver operating characteristic analysis, PC, PS, and ATIII could significantly predict PE and its subtypes at certain cutoffs with high accuracies (area under the curve [AUC] ≥0.70). Most women with PE are in a hypercoagulable state with lower natural anticoagulants. PC, PS, and ATIII are good predictive and diagnostic markers of PE and its subtypes (early-onset PE [EO-PE] and late-onset PE [LO-PE]) and should be explored in future studies.

Prevalence and risk factors of malaria and anaemia and the impact of preventive methods among pregnant women: A case study at the Akatsi South District in Ghana.

AIM: This study aimed to ascertain the prevalence and risk factors of malaria and anaemia as well as the impact of preventive methods among pregnant women at the Akatsi South District Hospital of Ghana. SUBJECTS AND METHODS: A hospital based cross-sectional study using simple random sampling technique was conducted among 200 pregnant women receiving antenatal care and laboratory services at the Akatsi District Hospital from May 2016 to July 2016. A semi-structured questionnaire was administered to obtain participants' malaria preventive methods in addition to demographic and gestational details. Participants' hemoglobin and malaria status were assessed using one milliliter (1 ml) whole blood collected from each participant following standard procedures. Factors that produced a p-value of ≤0.2 from the univariate model were included in the final model. Association between potential covariates and the outcomes was assessed using multivariate logistic regression. The Clopper-Pearson test statistic was used to determine the 95% confidence intervals of the outcome variables of interest. We also estimated the population attributable fraction (PAF) of anaemia due to malaria by substituting the adjusted relative risk estimates (RRi) (using the adjrr command in STATA) of anaemia due to malaria into the category-specific attributable formula. P-values of <0.05 were considered statistically significant. RESULTS: Prevalence of anaemia in pregnancy (AiP), malaria in pregnancy (MiP) and AiP/MiP comorbidity was 63.5% (95% CI:56.4-70.2), 11.0% (96% CI:7.0-16.2) and 10.5% (95% CI:6.6-15.6) respectively. Prevalence rates of AiP (66.7%) and MiP (18.5%) predominated among pregnant women aged < 20 years. PAF of AiP due to MiP was 34.5% (95% CI:23.8-43.6). High use of IPTp-SP, 64.0% (95% CI:56.9-70.6) and LLIN, 90.0% (95% CI:85.0-93.8) was observed in this study. Only 42.0% (95% CI:35.1-49.2) used repellent. Not being on the IPTp-SP program posed a 11.70 times risk of MiP (95% CI:2.32-58.96; p = 0.003) compared to pregnant women on the IPTp-SP program. Similarly, not sleeping under LLIN posed an 8.07 times risk of MiP (95% CI:1.98-32.2; p = 0.004) compared to pregnant women who slept under LLIN. Meanwhile, being positive for MiP posed a 12.10 times risk (95% CI:1.35-85.06; p = 0.025) of AiP compared to those negative for malaria whereas failure to attend ANC as scheduled posed 6.34 times risk (95% CI:1.81-22.19; p = 0.004) of AiP among the pregnant women studied. CONCLUSION: The prevalence of MiP and AiP among pregnant women in the Akatsi South District remains a great concern. High utilization of IPTp-SP and LLIN was observed with a resultant positive effect on malaria prevalence among pregnant women. Improved access to IPTp-SP and LLIN is hence encouraged to help further diminish the risk of malaria infection amongst pregnant women in the District.

Prevalence and Risk Factors of Preterm Birth Among Pregnant Women Admitted at the Labor Ward of the Komfo Anokye Teaching Hospital, Ghana.

Preterm birth is a global epidemic and a leading cause of neonatal mortality in Sub-Saharan Africa. We evaluated the prevalence and risk factors of preterm birth among women attending the labor ward for delivery at a tertiary hospital in Ghana. This comparative cross-sectional study was conducted among a cohort of 209 pregnant women admitted to the labor ward of the Komfo Anokye Teaching Hospital (KATH). Pregnant women who delivered between 28 and 36 completed weeks of gestation were classified as preterm delivery whereas those who delivered after 37-42 completed weeks were described as term. Sociodemographic, clinical, and obstetric data were collected from patient's folder and hospital archives. Categorical variables were analyzed and expressed as frequencies and proportions. We determined the association between obstetric factors and preterm delivery with multiple logistic regressions. Significance level of the strength of association was determined at p-value < 0.05. of the 209 participants, the prevalence of preterm birth was 37.3% (78/209) whereas 62.7% (131/209) delivered at Term. Intrauterine growth restriction (IUGR) [aOR = 2.15, 95% CI = (1.819.55), p = 0.0390], HELLP (hemolysis, elevated liver enzymes and low platelet count) syndrome [aOR = 3.94, 95% CI = (1.64-9.48), p = 0.0020], early gestational obesity [aOR = 2.11, 95% CI = (1.31-11.92), p = 0.0480] and preeclampsia [aOR = 4.56, 95% CI = (1.63-12.76), p = 0.004] were identified as independent risk factors of preterm birth. Prevalence of preterm birth was high among women attending labor admission at the Komfo Anokye Teaching Hospital and this was independently influenced by IUGR, HELLP syndrome, early gestational obesity, and preeclampsia. Identifying early signs of adverse pregnancy outcomes would inform the need for management policy to prevent high prevalence of preterm births.

Placental lesions and differential expression of pro-and anti-angiogenic growth mediators and oxidative DNA damage marker in placentae of Ghanaian suboptimal and optimal health status pregnant women who later developed preeclampsia.

BACKGROUND: Angiogenic growth mediators (AGMs) and oxidative stress (OS) both play essential roles in normal placental vascular development and as such, placental alterations in these factors contribute to pre-eclampsia (PE). Suboptimal health status (SHS), an intermediate between health and disease, has been associated with imbalanced AGMs and OS biomarkers. Thus, SHS pregnant women may be at increased risk of developing PE and may present abnormal placental alteration and expression of AGMs and OS compared to optimal health status (OHS) pregnant women. We examined the histopathological morphology, immunohistochemical expression of AGMs antibodies and oxidative DNA damage marker in the placentae of SHS and OHS pregnant women who developed early-onset PE (EO-PE) and late-onset (LO-PE) compared to normotensive pregnancy (NTN-P). METHODS: This nested case-control study recruited 593 singleton normotensive pregnant women at baseline (10-20 weeks gestation) from the Ghanaian Suboptimal Health Status Cohort Study (GHOACS) undertaken at the Komfo Anokye Teaching Hospital, Ghana. Socio-demographic, clinical and obstetrics data were collected, and a validated SHS questionnaire-25 (SHSQ-25) was used in classifying participants into SHS (n = 297) and OHS (n = 296). Participants were followed until the time of PE diagnosis and delivery (32-42 weeks gestation). Blood samples were collected at the two-time points and were assayed for AGMs; soluble fms-like tyrosine kinase-1 (sFlt-1), placental growth factor (PIGF), vascular endothelial growth factor-A (VEGF-A), and soluble endoglin (sEng), and OS biomarkers; 8-hydroxydeoxyguanosine (8-OHdG), 8-epiprostaglandinF2-alpha (8- epi-PGF2α) and total antioxidant capacity (TAC) using ELISA. Placental samples were collected for histopathological and immunohistochemical analysis. RESULTS: Of the 593 pregnant women, 498 comprising 248 SHS and 250 OHS women returned for delivery and were included in the final analysis. Of the 248 SHS women, 56, 97 and 95 developed EO-PE, LO-PE and NTN-P, respectively, whereas 14, 30 and 206 of the 250 OHS mothers developed EO-PE, LO-PE and NTN-P, respectively. At baseline, SHS_NTN pregnant women had a significant imbalance in AGMs and OS biomarkers compared to OHS_NTN pregnant women (p<0.0001). At the time of PE diagnosis, SHS_NTN-P women who developed EO-PE, LO-PE, and NTN-P had lower serum levels of P1GF, VEGF-A and TAC and correspondingly higher levels of sEng, sFlt-1, 8-epiPGF2α, and 8-OHdG than OHS-NTN-P women who developed EO-PE and LO-PE, NTN-P (p<0.0001). A reduced placental size, increased foetal/placental weight ratio, and a significantly higher proportion of fibrinoid necrosis, infarction, villous fibrin, syncytial knots, calcification, chorangiosis, tunica media/vascular wall hypertrophy and chorioamnionitis was associated with the SHS group who developed PE (EO-PE>LO-PE) more than OHS groups who developed PE (EO-PE>LO-PE) when all were compared to NTN-P (p<0.0001). The intensity of antibody expression of PIGF and VEGF-A were significantly reduced, whereas Flt-1, Eng and 8-OHdG were significantly increased in placentae from SHS-pregnant women who developed EO-PE>LO-PE more than OHS- pregnant women who developed EO-PE>LO-PE when all were compared to NTN-P (p<0.0001). CONCLUSION: Increased lesions, oxidative DNA damage, and imbalanced expression between pro-and anti-AGMs are associated more with SHS-embodied PE placentae rather than OHS-embodied PE subtypes, thus potentially allowing differential evaluation of PE.

Reference intervals for selected haematological and biochemical parameters among apparently healthy adults in different eco-geographical zones in Ghana.

BACKGROUND: Due to the influence of gender, race/genetics, age, lifestyle habits and geography on the references intervals (RIs), the Clinical and Laboratory Standards Institute (CLSI) recommends the determination of population-specific RIs. Ghana continues to depend on pre-established RIs from other countries which poses the risk of misdiagnoses and wrong treatment. This study presents the haemato-biochemical RIs from four eco-geographical zones in Ghana. METHODS: In this population-based cross-sectional study, a total of 1227 randomly selected healthy voluntary blood donors from the four eco-geographic zones (Coastal Savannah, Rain Forest, Savannah and Transitional) were enrolled and screened. Based on the CLSI Guidance Document C28A2992, the data of eligible participants were used to non-parametrically determine the RIs for the haemato-biochemical parameters at the 2.5th and 97.5th percentiles. Comparison of analytes by gender was done by Wilcoxon rank sum test and eco-geographic differences were assessed using the Kruskal-Wallis with the Dunn post hoc multiple comparison tests. RESULTS: There were statistically significant differences in most of the haematological parameters (RBC, Hb, HCT, MCV, PLT, WBC; p-values <0.0001 and MCH; p-value = 0.007), and biochemical analytes (Urea, Cr, Trig, HDL-C, AST, ALT, ALP, GGT, BID, BIT, Prot-T and Albumin; p-values <0.0001) based on gender. Significant inter eco-geographic (intra-population) variations and substantial differences between the established RI and the RIs accompanying the analyzers used were also observed. CONCLUSION: This study reports significant inter-sex and inter-geographical differences in haemato-biochemical RIs in Ghana as well as differences in RIs with both the RIs accompanying the analyzers and those of other countries. Determining RIs representative of populations and including them in the report systems of laboratories to ensure effective and efficient healthcare service delivery is thus recommended.

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.

Acute Phase Responses Vary Between Children of HbAS and HbAA Genotypes During Plasmodium falciparum Infection.

PURPOSE: Haemoglobin genotype S is known to offer protection against Plasmodium falciparum infections but the mechanism underlying this protection is not completely understood. Associated changes in acute phase proteins (APPs) during Plasmodium falciparum infections between Haemoglobin AA (HbAA) and Haemoglobin AS (HbAS) individuals also remain unclear. This study aimed to evaluate changes in three APPs and full blood count (FBC) indices of HbAA and HbAS children during Plasmodium falciparum infection. METHODS: Venous blood was collected from three hundred and twenty children (6 months to 15 years) in Begoro in Fanteakwa District of Ghana during a cross-sectional study. Full blood count (FBC) indices were measured and levels of previously investigated APPs in malaria patients; C-reactive protein (CRP), ferritin and transferrin measured using Enzyme-Linked Immunosorbent Assays. RESULTS: Among the HbAA and HbAS children, levels of CRP and ferritin were higher in malaria positive children as compared to those who did not have malaria. The mean CRP levels were significantly higher among HbAA children (p=0.2e-08) as compared to the HbAS children (p=0.43). Levels of transferrin reduced in both HbAA and HbAS children with malaria, but the difference was only significant among HbAA children (p=0.0038), as compared to the HbAS children. No significant differences were observed in ferritin levels between HbAA and HbAS children in both malaria negative (p=0.76) and positive (p=0.26) children. Of the full blood count indices measured, red blood cell count (p=0.044) and haemoglobin (Hb) levels (p=0.017) differed between HbAA and HbAS in those without malaria, with higher RBC counts and lower Hb levels found in HbAS children. In contrast, during malaria, lymphocyte and platelet counts were elevated, whilst granulocytes and Mean Cell Haematocrit counts were reduced among children of the HbAS genotypes. CONCLUSION: Significant changes in APPs were found in HbAA children during malaria as compared to HbAS children, possibly due to differences in malaria-induced inflammation levels. This suggests that the HbAS genotype is associated with better control of P. falciparum infection-induced inflammatory response than HbAA genotype.