Browsing by Author "Sakyi, Samuel Asamoah"

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    Anti-cyclic Citrullinated Peptide as an early and accurate laboratory marker for the diagnosis of Rheumatoid Arthritis (RA) and the prevalence of HLA-B27 among Ankylosing Spondylitis patients in Ghana
    (2010) Sakyi, Samuel Asamoah
    Rheumatoid Arthritis (RA) and Ankylosing Spondylitis (AS) are two of the most common autoimmune diseases. This study aimed to obtain the best biomarker for early and accurate diagnosis of RA, determine the prevalence of HLA-B27 among AS patients in Ghana, and examine if there is dyslipidaemia and oxidative stress among them. A total of one hundred and sixty-seven (167) subjects were recruited for this cross sectional study from October 2006 to June 2008. They were made up of one hundred and two (102) RA and sixty-five (65) AS subjects. Both groups were recruited from the orthopaedic departments of Komfo Anokye Teaching Hospital (KATH), Korle-Bu Teaching Hospital (KBTH) and from subjects visiting MEDILAB centres nationwide with laboratory requests to investigate rheumatoid conditions. Those who were confirmed with either RA or AS and consented were recruited. Fifty (50) healthy blood donors with similar age and sex distribution as the subjects were recruited as control group. After ethical approval and informed consent had been obtained, information on socio-demographic characteristics and medical history were obtained from standardized questionnaires which was administered to them and through their medical records. Rheumatoid Factor (RF)-latex and its isotypes (RF-IgA and RF-IgM) were assayed and compared with anti-CCP as specific markers for early diagnosis of RA in Ghana. HLA-B27 was assayed to determine its prevalence among AS subjects. Lipid profile, Tumour Necrosis Factor-α (TNF-α) and Erythrocyte Sedimentation Rate (ESR) were estimated to determine the presence of dyslipidaemia and inflammation among AS and RA subjects. To evaluate the presence of oxidative stress, MDA and vitamin C were also determined among RA and AS subjects. Out of 102 RA subjects, Sensitivity was highest for RF-IgM test (58.45%) followed by anti-CCP antibody (54.34%), RF-latex (52.48%) and RF-IgA tests (28.51%). Specificity of 96.67% was obtained for anti-CCP, 69.52% for RF-IgM, 63.12% for RF-latex and 51.75% for RF-IgA. RA and AS subjects had a higher mean level of total cholesterol (TC) 5.88 ± 0.06 mmol/l. The difference between the RA and AS subjects’ cholesterol level and that of the control was significant (p<0.001) with a significant (r2= 0.0619, p= 0.002) positive correlation between serum triglycerides levels of AS and RA patients and that of the control. RA and AS subjects exhibited a slightly higher mean level of triglycerides 0.88 ± 0.16 mmol/l as compared to 0.56±0.05 mmol/l of controls. The difference between the subjects triglycerides levels and that of the control was significant (p<0.001). The mean HDL level for the patients was significantly lower (p<0.001) as compared to that of the control. The mean LDL level indicates a significantly higher (p<0.001) value as compared to that of the controls with a significant (r2= 0.0619, p= 0.002) positive correlation between serum triglycerides, total cholesterol, HDL and LDL of RA subjects. A significant (p<0.001) atherogenic ratios i.e. TC/HDL-C and LDL-C/HDL-C were observed among the subjects and controls. Out of 65 AS subjects examined there were four (4) HLA-B27 positives representing 4.6%, three (3) of the HLA-B27 positives were males, the mean Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) score was 44.7/100. Forty-eight (48) AS patients had sacroilitis in their x-ray reports. None had a family history of AS or any extra-articular manifestations. TNF-α level among RA and AS patients were 11.87 ± 0.30 pg/ml and 13.11 ± 0.50 pg/ml respectively as compared to normal control of 5.70 ± 0.48 pg/ml whiles the mean ESR for AS was 34.64 ± 1.87 mm/hr and that of RA was 24.67 ± 1.87 mm/hr as compared to 9.23 ± 0.91 mm/hr of controls. The mean MDA level for AS subjects was 0.75 ± 0.03 mmol/l and that of RA subjects was 1.57± 0.06µmol/l as compared to normal controls of 0.58 ± 0.02mmol/l. MDA levels had a significant correlation with the serum levels of vitamin C. Vitamin C was 0.85 ± 0.03mmol/l for AS subjects and 0.95 ± 0.02mmol/l for RA subjects as compared to 1.12± 0.01µmol/l of control. This study has demonstrated that anti-CCP has the highest specificity and that the combination of anti-CCP and RF-IgM assay are highly specific and moderately sensitive for diagnosing RA, making this combination of autoantibodies a powerful tool in the serologic assessment of RA in Ghana. The presence of anti-CCP at disease onset means that they have a high positive predictive value of 100, a negative predictive value of 22.6 with 4.98 and 0.68 as positive and negative likelihood ratios respectively, predict the development of erosive joint lesions and the detection of these antibodies can therefore be used in clinical practice to help plan a therapeutic strategy. Findings from this study further showed that HLA-B27 is present in some AS patients. The BASDAI scores and other clinical features clearly denote the presence of the disease with moderate disease activity. Dyslipidaemia is present in early RA and AS subjects, and that proinflammation and inflammation markers plays a pivotal role in the development of atherosclerosis as evidenced in the increase in TNF-α, ESR and a more atherogenic lipid profile, thereby increasing cardiovascular risk. Increased oxidative stress is evidenced by raised MDA and decreased vitamin C and that lipid peroxidation is a giant distracter in RA and AS.
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    Demographic, Clinical Profile of Rheumatoid Arthritis Patients and Their Association with Disease Severity in Ghana
    (Hindawi, 2024) Buckman, Tonnies Abeku; Sakyi, Samuel Asamoah; Yeboah-Mensah, Kwame; Antwi, Maxwell Hubert; Darban, Isaac; Owusu-Brenya, Lawrence; Yorke, Joseph; 0000-0002-5229-0340
    Background. Rheumatoid arthritis (RA) is one of the frequent chronic, systemic, inflammatory autoimmune disorders with an estimated global prevalence of 1%. RA leads to joint destruction and disability if left untreated. Ghana has seen very few studies on RA, and little is known about the disease’s severity and related variables. This study sought to characterize the clinical presentation and determine disease severity and associated risk factors with disease severity among RA patients in a tertiary hospital in Ghana. Methods. This cross-sectional study was conducted between September 2020 and August 2021. This study included 56 consecutively consenting RA patients from the Komfo Anokye Teaching Hospital orthopaedic unit. Diagnosis of RA was based on the updated American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) 2022 rheumatoid arthritis classification criteria by a rheumatologist. A study questionnaire was used to gather participant demographics and clinical features, and results from the laboratory were taken from the patients’ charts and medical records. The patients’ disease severity was evaluated based on the rheumatoid arthritis disease activity score, which is based on a 28-joint count (DAS28), and their functioning was evaluated using the modified health assessment questionnaire. Results. The participants’ mean age was 51 25 ± 13 22 years. Out of the total participants, 46 were females, and 10 were males (female-to-male ratio 4.6 : 1). Moreover, 37.50% had arthritis of the hand; 5.30% had severe disease, and 94.60% were not severe. A majority (76.80%) were on methotrexate medication. The most frequently involved joints were the knee (42.90%), wrist (32.10%), and elbow (12.50%). There was no statistically significant association with disease severity and a functional status score of >0.5 (cOR: 10.60, 95% CI (0.52-217.30); p = 0 124). In addition, marital status (p = 0 04), disease duration (p = 0 04), family complaints (p = 0 02), and ESR (p = 0 03) were significantly associated with disease severity. Conclusion. RA is predominant among elder populations and females. Disease duration, family complaints, and ESR are associated with disease severity. The findings of this study call for interventions towards ensuring early diagnosis of RA among high-risk populations to enhance good management practices.
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    Investigation of serum level relationship of pro-inflammatory and anti-inflammatory cytokines with vitamin D among healthy Ghanaian population
    (BMC Publisher, 2024) Antwi, Maxwell Hubert; Sakyi, Samuel Asamoah; Appiah, Seth Christopher Yaw; Buckman, Tonnies Abeku; 0000-0002-5229-0340
    Background The interplay between vitamin D status and inflammatory cytokines in a supposedly sufficient sunshine environment has not well been evaluated. The study sought to determine their association. Methods This cross-sectional study involved 500 healthy adult blood donors from some selected hospitals in Ghana enrolled from June to November 2016. Venous blood samples were obtained from participants, 25(OH)D, TNF-alpha, IFN-gamma, and IL 10 were measured using enzyme linked immunosorbent assay (ELISA) technique. Serum levels of 25(OH)D<20ng/ml were classified as being deficient or low. Results The average age of the participants was 27.97 years. No statistically significant association was established between 25(OH) D status, mean age (p=0.1693), and gender (p=0.5461) of study participants. Similarly, the median 25(OH) D (p=0.8392), IL-10 (p=0.5355), TNF-alpha (p=0.9740), and IFN-gamma (p=0.6908) were not significantly different across gender. There was a significantly increased levels of TNF-alpha (p<0.0001) and IFN-gamma (p<0.0001) among participants with 25(OH) D deficiency compared to those without deficiency. Concurrently, participants with 25(OH)D deficiency had a significantly reduced levels of IL-10 (p<0.0001) compared to those without 25 (OH) D deficiency. The most accurate biochemical markers for identifying 25 (OH) D deficiency were IFN gamma (AUC=0.879; p<0.0001) followed by TNF-gamma (AUC=0.849; p<0.0001) and IL-10 (AUC=0.707; p<0.0001). Conclusion There was a significant association between vitamin D levels and pro-inflammatory cytokines (TNF alpha, IFN-gamma) and anti-inflammatory cytokine (IL 10) among healthy Ghanaian populace.
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    Molecular characterization of interactions between the D614G variant of SARS-CoV-2 S-protein and neutralizing antibodies: A computational approach
    (Infection, Genetics and Evolution, 2021) Kwarteng, Alexander; Asiedu, Ebenezer; Sylverken, Augustina Angelina; Larbi, Amma; Sakyi, Samuel Asamoah; Asiedu, Samuel Opoku; 0000-0002-0893-2908; 0000-0003-2867-1984; 0000-0002-7691-914X; 0000-0002-3814-6924; 0000-0001-5168-4762; 0000-0002-0352-3195
    The D614G variant of SARS-CoV-2 S-protein emerged in early 2020 and quickly became the dominant circulating strain in Europe and its environs. The variant was characterized by the higher viral load, which is not associated with disease severity, higher incorporation into the virion, and high cell entry via ACE-2 and TMPRSS2. Previous strains of the coronavirus and the current SARS-CoV-2 have demonstrated the selection of mutations as a mechanism of escaping immune responses. In this study, we used molecular dynamics simulation and MM-PBSA binding energy analysis to provide insights into the behaviour of the D614G S-protein at the molecular level and describe the neutralization mechanism of this variant. Our results show that the D614G S-protein adopts distinct conformational dynamics which is skewed towards the open-state conformation more than the closed-state conformation of the wild-type S-protein. Residue-specific variation of amino acid flexibility and domain specific RMSD suggest that the mutation causes an allosteric conformational change in the RBD. Evaluation of the interaction energies between the S-protein and neutralizing antibodies show that the mutation may enhance, reduce or not affect the neutralizing interactions depending on the neutralizing antibody, especially if it targets the RBD. The results of this study have shed insights into the behaviour of the D614G S-protein at the molecular level and provided a glimpse of the neutralization mechanism of this variant.
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    Rapid Extraction Method of Mycobacterium ulcerans DNA from Clinical Samples of Suspected Buruli Ulcer Patients
    (Diagnostics, 2019-11-26) Phillips, Richard Odame; Frimpong, Michael; Ahor, Hubert Senanu; Sakyi, Samuel Asamoah; Agbavor, Bernadette; et. al
    Isothermal amplification techniques such as recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP) for diagnosing Buruli ulcer, a necrotic skin disease caused by Mycobacterium ulcerans, have renewed hope for the molecular diagnosis of clinically suspected Buruli ulcer cases in endemic districts. If these techniques are applied at district-level hospitals or clinics, they will help facilitate early case detection with prompt treatment, thereby reducing disability and associated costs of disease management. The accuracy as well as the application of these molecular techniques at point of need is dependent on simple and fast DNA extraction. We have modified and tested a rapid extraction protocol for use with an already developed recombinase polymerase amplification assay. The entire procedure from “sample in, extraction and DNA amplification” was conducted in a mobile suitcase laboratory within 40 min. The DNA extraction procedure was performed within 15 min, with only two manipulation/pipetting steps needed. The diagnostic sensitivity and specificity of this extraction protocol together with M. ulcerans RPA in comparison with standard DNA extraction with real-time PCR was 87% (n = 26) and 100% (n = 13), respectively. We have established a simple, fast and e cient protocol for the extraction and detection of M. ulcerans DNA in clinical samples that is adaptable to field conditions.