Respiratory Burst Factors in Nigerian Patients with COVID-19

Respiratory burst function resulting in the release of reactive oxygen species from leucocytes is one of the key mechanisms of innate immune system to prevent the establishment of intracellular pathogens in the host cells. Previous studies on COVID-19 patients concentrated on adaptive immunity while study on respiratory burst functions is lacking. Respiratory burst mediators levels [nitric oxide (NO) and hydrogen peroxide (H2O2)] and respiratory burst enzymes activities [Catalase (CAT), Myeloperoxidase (MPO) and Superoxide dismutase (SOD)] were quantitated in the plasma Mean plasma NO level, MPO activity and H2O2 level were significantly decreased while SOD activity was significantly increased in COVID-19 patients at admission compared with control. Mean plasma NO level significantly decreased while MPO activity was significantly increased in COVID-19 patients at discharge compared with control. Plasma NO level, H2O2 level and MPO activity were significantly increased in COVID-19 patients at discharge compared with COVID-19 patients at admission. In COVID-19 patients that spent ≥10days in admission, the levels of NO and H2O2 were significantly increased compared with the levels of NO and H2O2 in COVID-19 patients that spent <10days in admission. In male COVID-19 patients, NO level and MPO activity were significantly increased compared with MPO activity in female patients. In COVID-19 patients ≥40years of age, NO level was significantly decreased while MPO activity was significantly increased compared with COVID-19 patients <40yrs of age. In male COVID-19 patients, NO level and MPO activity was significantly increased compared with MPO activity in female patients. It could be concluded from this study that factors of respiratory burst which are components of the innate immune system are altered in COVID-19 patients and could be involved in the immune-pathogenecity of SARS-CoV-2; and that MPO coupled with NO may explain differential severities of COVID-19 among genders and age groups

Antioxidant Vitamins Are Correlated with Different Aspects of Phagocytic Processes in Healthy Nigerians: Benefits As Supplements During Antimicrobial Treatment

Background: Antioxidant vitamins are important for the immune system to function efficiently through several mechanisms. However, according to several previous studies, individual step of leucocyte phagocytosis is not correlated with different antioxidant vitamins. Methods: This study included 50 healthy Nigerians whose cellular phagocytic mechanism such as percentage leucocyte migration (%LM) and intracellular killing (%Nitroblue Tetrazolium Test) were determined by microscopy, neutrophil chemokines [plasma interleukin 8 (IL-8)] was determined using ELISA, and respiratory burst indices [plasma catalase (CAT), superoxide dismutase (SOD), myeloperoxidase (MPO), hydrogen peroxide (H2O2), and nitric oxide (NO)] were determined by spectrophotometry. While the plasma antioxidant vitamins (Vitamins A, C, and E) were determined using HPLC, the phagocytic indices, chemokines, and respiratory burst indices were correlated with plasma antioxidant vitamins using Spearman’s Correlation analysis at α0.05. Results: The results show that although among the healthy Nigerian adults, vitamin C was significantly and positively correlated with %NBT, it was negatively correlated with CAT activity. Vitamin A showed a significantly positive correlation with SOD while Vitamin E showed a significantly negative correlation with MPO. Conclusions: These findings suggest that antioxidant vitamins affect different stages of phagocytosis. It is advisable to use a combination of antioxidant vitamins as supplements with recommended treatment strategies against intracellular micro-organisms or inflammatory diseases. Keywords: Antioxidants, Intracellular microbial killing, Vitamin

Human Beta Defensin 1 (hbd1) Levels in Sputum and Lysate of Mononuclear Blood Cells of Drug-Sensitive and Drug-Resistant Pulmonary Tuberculosis Patients Attending a Tertiary Hospital in Ibadan, Nigeria.

Background: Mycobacterium tuberculosis (M. tb) that causes pulmonary tuberculosis (PTB) occupies the lungs, while human β-defensin-1 (hBD1) is expressed in all human epithelial tissues as one of the products of phagocytic leucocytes, especially at the site of microbial colonisation such as the lungs. The involvement of hBD1 in mycobacterial infection has not been extensively studied, thus there is the need to measure the levels of the hBD1 in mononuclear cell lysates and sputum of PTB patients at diagnosis. Materials and Methods: Ninety participants aged between 15 and 64 years were recruited as follows: 30 newly diagnosed multi-drug-resistant TB (MDR-TB) patients and 30 newly diagnosed drug-sensitive TB patients (DS-TB) from MDR-TB Treatment centre and the Medicine Outpatient Clinic at University College Hospital (UCH) Ibadan, Nigeria. Thirty (30) non-TB apparently healthy individuals served as controls. The analytical method employed for the measurement of hBD1 in the sputum and lysate was the Enzyme-linked immunosorbent assay (ELISA). The data were expressed as mean and standard deviation, and the differences between the means were established using Student (t) test. P-value ≤ 0.05 indicated statistical significance. Results: The mean levels of lysate and sputum hBD1 were not significantly different in newly diagnosed DS–TB patients (D0)compared with control(p > 0.05). Whereas, the mean levels of lysate and sputum hBD1 were significantly higher in newly diagnosed MDR–TB patients (M0) compared with newly diagnosed DS–TB patients (D0)or control (p < 0.05). Conclusion: Due to the higher levels of hBD1 in the sputum and lysate of M0 than in D0, one might conclude that there is a relationship between chronicity of PTB and hBD1 level

Innate Cellular Immunity in Newly Diagnosed Pulmonary Tuberculosis Patients and During Chemotherapy

Background: Leukocyte migration (LM) and intracellular killing aspects of the innate immune responseplay important roles in protection against and containment and cure of Mycobacterium tuberculosis infection, and thus may be exploited as immunotherapeutic targets to improve the management and treatment outcomes of patients with tuberculosis (TB). Objectives: The aim of this study was to assess LM and mediators of intracellular killing in patients with TB at the time of diagnosis and during anti-TB chemotherapy and compare them with apparently healthy controls. Methods: We recruited 24 patients who were newly diagnosed with pulmonary TB and 20 apparently healthy individuals. Blood was drawn from patients with TB at the time of diagnosis, and after 2, 4, and 6 months of anti-TB chemotherapy and control. In vitro percentage LM (%LM) upon stimulation with Bacillus Calmette-Guérin vaccine, percentage nitroblue tetrazolium (%NBT) reduction, plasma concentrations of hydrogen peroxide (H2O2), and nitric oxide(NO) were assessed in both groups. Findings: Percentage NBT was significantly reduced in patients with TB at 2 months of anti-TB chemotherapy compared with patients at diagnosis and in healthy controls, whereas %LM was significantly increased in patients at 4 months of anti-TB chemotherapy compared with patients at diagnosis and controls. Mean plasma H2O2 and NO were significantly reduced in patients at diagnosis and throughout the period of anti-TB chemotherapy compared with the control group. Significant decreases were demonstrated in mean plasma H2O2 and NO in patients at 2 and 4 months of anti-TB chemotherapy, respectively, compared with patients at diagnosis. There was significant positive correlation between %NBT with plasma H2O2 and NO, but %LM was negatively correlated with plasma H2O2 in this group. Conclusion: The intracellular killing aspect of innate cellular immunity is deficient in patients with TB, especially 2 to 4 months after commencement of treatment. Therefore, measures (eg, arginine supplementation) to improve intracellular killing in these patients is advocated. Moreover, %LM assay with Bacillus Calmette-Guérin vaccine as an antigen may be used to differentiate those newly diagnosed patients from those on anti-TB chemotherapy

Thyroid function in multidrug-resistant tuberculosis patients with or without human immunodeficiency virus (HIV) infection before commencement of MDR-TB drug regimen.

Background: Mycobacterium tuberculosis and human immunodeficiency virus (HIV) are known to cause abnormal thyroid function. There is little information on whether HIV infection aggravates alteration of thyroid function in patients with MDRTB. Objectives: This study was carried out to determine if HIV co-infection alters serum levels of thyroid hormones (T3, T4) and thyroid stimulating hormone (TSH) in patients with MDR-TB patients and to find out the frequency of subclinical thyroid dysfunction before the commencement of MDR-TB therapy. Methods: This observational and cross-sectional study involved all the newly admitted patients in MDR-TB Referral Centre, University College Hospital, Ibadan, Nigeria between July 2010 and December 2014. Serum levels of thyroid stimulating hormone (TSH), free thyroxine (fT4) and free triiodothyronine (fT3) were determined using ELISA. Results: Enrolled were 115 patients with MDR-TB, out of which 22 (19.13%) had MDR-TB/HIV co-infection. Sick euthyroid syndrome (SES), subclinical hypothyroidism and subclinical hyperthyroidism were observed in 5 (4.35%), 9 (7.83%) and 2 (1.74%) patients respectively. The median level of TSH was insignificantly higher while the median levels of T3 and T4 were insignificantly lower in patients with MDR-TB/HIV co-infection compared with patients with MDRT-TB only. Conclusion: It could be concluded from this study that patients with MDR-TB/HIV co-infection have a similar thyroid function as patients having MDR-TB without HIV infection before commencement of MDR-TB drug regimen. Also, there is a possibility of subclinical thyroid dysfunction in patients with MDR-TB/HIV co-infection even, before the commencement of MDR-TB therapy

Serum Micronutrients in Helminth-infected Pregnant Women and Children: Suggestions for Differential Supplementation During Anti-helminthic Treatment

Background: The prevalence of helminth infection, which is known to affect nutritional status of the host, varies with age. The complex interplay between ages, nutrient requirements, and infection necessitated the need to recommend micronutrient supplementation during helminth infection among different age groups. Objective: The aim of this study was to determine the pattern of alteration in selected micronutrients in pregnant women and preschool- and school-aged children with helminth infection. Methods: We screened 245 pregnant women and 349 children for helminth infection. Of these, 17 (6.9%) pregnant women and 102 (29.2%) children (42 preschool- and 60 school-aged) had helminth infection. Only 'Ascaris lumbricoides' was found in pregnant women, whereas the children had 'A lumbricoides', hookworm, 'Fasciola hepatica', and 'Trichuris' trichiura infections. The helminth-infected (HI) pregnant women, preschool-aged children, and school-aged children were matched with helminth-negative (HN) pregnant women (n = 21), preschool-aged children (n = 42), and school-aged children (n = 50) who served as controls. Venous blood samples were obtained and analyzed for iron (Fe), zinc (Zn), selenium (Se), and vitamins A and C. Statistical analysis was done using Student’s 't' test, and 'P' vitamin A were significantly lower in the HI than in the HN group. Similarly, serum levels of Zn and vitamin A were significantly lower in HI school-aged children than in the HN group. However, serum levels of Se were significantly higher in HI children (both age groups) than in the corresponding HN group. Conclusion: Helminth infection alters different types of micronutrients in children and pregnant women. Results from the present study therefore suggest monitoring Fe, Zn, or vitamin A supplementation with an anti-helminthic regimen

Serum Levels of Cytokines and IgE in Helminth-Infected Nigerian Pregnant Women and Children

Background: Helminth infection is an important health challenge. Because of modulation of the immune response toward T-helper 2 (Th2) cells, the immunologic interplay that manifest during the coexistence of helminth infection with other conditions is still poorly understood. Objective: This study determined the pattern of alteration in selected cytokines and immunoglobulin E(IgE) in pregnant women, preschool aged children, and school-aged children with helminth infection compared with uninfected groups. Methods: Seventeen pregnant women, 42 preschool-aged children, and 60 school-aged children with helminth infection (HI) were recruited into this study. They were matched with 21 pregnant women, 42 preschool-aged children, and 50 school-aged children without helminth infection (HN) who served as controls. Venous blood samples were collected from each participant and analyzed for serum levels of tumor necrosis factor α (TNF-α), interleukin-10 (IL-10), interleukin-8 (IL-8), interleukin-6 (IL-6), and IgE. Statistical analysis was done using the Student 't' test, and 'P' < .05 was considered as statistically significant. Findings: Only serum level of IgE was significantly elevated in HI pregnant women compared with HN pregnant women. In HI preschool- and school-aged children, serum levels of IL-8, IL-6, and IgE were significantly elevated compared with HN children. However, preschool- and school-aged children with HI had similar levels of serum TNF-α and IL-10 compared with their corresponding HN groups. Conclusions: It could be concluded that altered cytokines expression in children and pregnant women with helminth infection might have some implications on need for deworming programs to improve pregnancy outcomes and vaccine responses

Development and Validation of an RP-HPLC Method for the Quantitative Analysis of Triclosan in Human Urine

Triclosan (TCS), a synthesized chlorinated phenolic compound, is commonly utilized in consumable products as an antimicrobial agent. TCS has sparked widespread awareness because of its toxicity and possible negative effect on public health in recent years. In this study, a highly sensitive, fast, and cost-effective isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method coupled with solid-phase extraction for analysis of triclosan in human urine samples was developed. The method utilized methanol and water in a ratio of 90 : 10 as the mobile phase on a Phenomenex Luna 3 µm C18(2) 100 Å, 150 × 4.60 mm stationary phase, with a runtime of 5 minutes. The method showed good resolution of triclosan in the presence of the sample matrix. Validation of the method was performed according to the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH). Linearity was tested over a range of 0.00625 µg/mL to 6.4 µg/mL, as accuracy recorded a recovery of 89.25%, 91.0%, and 92.75%. Limits of detection (LOD) and quantification (LOQ) were obtained to be 0.0173 µg/mL and 0.0525 µg/mL, respectively. The method proved to be robust over a temperature range of 26°C, 30°C, and 35°C and a flow rate of 0.5 ml, 1.0 ml, and 1.5 ml. The developed method was employed to detect and quantify triclosan in 153 urine samples, comprising 60 samples from Ibadan, Nigeria, and 93 samples from Kumasi, Ghana. Triclosan was detected in a total of 52 samples with an average content of 0.054588 µg/ml. This method can therefore be used for the routine analysis of triclosan in urine samples

Serum levels of anti-corona virus specific-IgG and -IgM antibodies in COVID-19 patients at admission and at discharge

Introduction. Clear understanding of duration of antibody based protective immunity following natural infection with SARSCoV-2 will give idea about the efficacy of proposed prophylactic vaccines against SARS-CoV-2, establishment of herd immunity and use of convalescent plasma. Aim. This study clarified the kinetics and magnitude of the initial antibody response against SARS-CoV-2 in a cohort of symptomatic COVID-19 patients from Ibadan, Nigeria. Material and methods. This study quantified immunoglobulin M (IgM) and G (IgG) antibodies recognizing the SARS-CoV-2 Spike (S) protein in 35 symptomatic COVID-19 patients at admission and at discharge using ELISA. Results. CovIgG was positive in none (0)% and 20% of COVID-19 patients at admission and at discharge respectively while CovIgM was positive in 54% and 69% of COVID-19 patients at admission and at discharged respectively. The level of CovIgG was significantly higher in COVID-19 patients at discharge compared with the level at admission while the level of CovIgM was insignificantly reduced in COVID-19 patients at discharge compared with the level at admission. Conclusion. The data indicates increased anti-SARS-COV-2 IgG Spike antibody in symptomatic COVID-19 at discharge, thus providing basis for antibody-based therapies to treat COVID-19 patient

Haematological changes and metabolic alterations in SARS-CoV-2 infected patients hospitalised at an Infectious Diseases Center, Ibadan, Nigeria

Introduction and aim. Following the use of repurposing drugs to successfully manage coronavirus disease 2019 (COVID-19) patients in an Infectious Diseases Center (IDC) in Nigeria, it was imperative to assess haematological changes and metabolic alterations in these patients which may inform recommendations for future use. Material and methods. Blood samples of admitted COVID-19 Nigerian patients during therapeutic management were analysed for haematological- (total white blood cells (WBC), lymphocyte, monocyte, neutrophil, eosinophil, basophil and neutrophil:lymphocyte ratio) and blood chemistry- parameters [total protein, total and conjugated bilirubin, alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine transaminase (ALT), gamma-glutamyl transferase (GGT), albumin, urea, creatinine, total cholesterol, triglycerides, low-density lipoprotein (LDL), high-density lipoprotein (HDL), PO₄³⁻, Ca²⁺, uric acid, Na⁺, K⁺, Cl⁻ and HCO₃⁻] using autoanalysers. The percentages of patients having values below, within and above reference ranges were compared using Chi-square test while the mean values at admission were compared with mean values at discharge using Student t-test. Results. The mean values of total protein, albumin, Na⁺, HCO₃⁻, uric acid, Ca²⁺, WBC, platelets, lymphocytes, eosinophils and basophils were significantly increased in COVID-19 patients at discharge compared with COVID-19 patients at admission. Also, more percentages of COVID-19 patients at discharge compared with COVID-19 patients at admission had albumin, ALP, total bilirubin, HDL, Na⁺, K⁺, Cl⁻, HCO₃⁻, urea, creatinine, WBC, lymphocytes, neutrophils, monocytes, eosinophils and basophils within normal reference intervals. Conclusion. This study showed that most metabolic and haematological derangements were normalised by repurposing drugs in most of the COVID-19 patients at this IDC, thus supporting the continuous use of this therapeutic option