Immune responses to P falciparum antibodies in symptomatic malaria patients with variant hemoglobin genotypes in Ghana.

BACKGROUND: Haemoglobin (Hb) variants such as sickle cell trait (SCT/HbAS) play a role in protecting against clinical malaria, but little is known about the development of immune responses against malaria parasite (Plasmodium falciparum surface protein 230 (Pfs230) and Plasmodium falciparum erythrocyte binding antigen 175 region-3 (PfEBA175-3R)) and vector (on the An. gambiae Salivary Gland Protein-6 peptide 1 (gSG6-P1)) antigens in individuals with variants Hb genotypes. This study assessed antibody (IgG) responses against malaria parasite, Pfs230 and PfEBA175-3R and vector, gSG6-P1 in febrile individuals with variant Hb genotypes. METHODS: The study was conducted on symptomatic malaria patients attending various healthcare facilities throughout Ghana. Microscopy and ELISA were used to determine the natural IgG antibody levels of gSG6-P1, PfEBA175-3R & Pfs230, and Capillarys 2 Flex Piercing was used for Hb variants determination. RESULTS: Of the 600 symptomatic malaria patients, 50.0% of the participants had malaria parasites by microscopy. The majority 79.0% (398/504) of the participants had Hb AA, followed by HbAS variant at 11.3% (57/504) and HbAC 6.7% (34/504). There were significantly (p < 0.0001) reduced levels of gSG6-P1 IgG in individuals with both HbAC and HbAS genotypes compared to the HbAA genotype. The levels of gSG6-P1 IgG were significantly (p < 0.0001) higher in HbAS compared to HbAC. Similarly, Pfs230 IgG and PfEBA-175-3R IgG distributions observed across the haemoglobin variants were significantly higher in HbAC relative to HbAS. CONCLUSION: The study has shown that haemoglobin variants significantly influence the pattern of anti-gSG6-P1, Pfs230, and PfEBA-175 IgG levels in malaria-endemic population. The HbAS genotype is suggested to confer protection against malaria infection. Reduced exposure to infection ultimately reduces the induction of antibodies targeted against P. falciparum antigens.

Comparison of microscopic and xpert MTB diagnoses of presumptive mycobacteria tuberculosis infection: retrospective analysis of routine diagnosis at Cape Coast Teaching Hospital.

INTRODUCTION: Tuberculosis is a global health problem that causes 1. 4 million deaths every year. It has been estimated that sputum smear-negative diagnosis but culture-positive pulmonary TB diagnosis contribute to 12.6% of pulmonary TB transmission. TB diagnosis by smear microscopy smear has a minimum detection limit (LOD) of 5,000 to 10,000 bacilli per milliliter (CFU/ml) of sputum result in missed cases and false positives. However, GeneXpert technology, with a LOD of 131-250 CFU/ml in sputum samples and its implementation is believe to facilitate early detection TB and drug-resistant TB case. Since 2013, Ghana health Service (GHS) introduce GeneXpert MTB/RIF diagnostic in all regional hospitals in Ghana, however no assessment of performance between microscopy and GeneXpert TB diagnosis cross the health facilities has been reported. The study compared the results of routine diagnoses of TB by microscopy and Xpert MTB from 2016 to 2020 at the Cape Coast Teaching Hospital (CCTH). METHODS: The study compared routine microscopic and GeneXpert TB diagnosis results at the Cape Coast Teaching Hospital (CCTH) from 2016 to 2020 retrospectively. Briefly, sputum specimens were collected into 20 mL sterile screw-capped containers for each case of suspected TB infection and processed within 24 h. The samples were decontaminated using the NALC-NaOH method with the final NaOH concentration of 1%. The supernatants were discarded after the centrifuge and the remaining pellets dissolved in 1-1.5 ml of phosphate buffer saline (PBS) and used for diagnosis. A fixed smears were Ziehl-Neelsen acid-fast stain and observed under microscope and the remainings were used for GeneXpert MTB/RIF diagnosis. The data were analyze using GraphPad Prism. RESULTS: 50.11% (48.48-51.38%) were females with an odd ratio (95% CI) of 1.004 (0.944-1.069) more likely to report to the TB clinic for suspected TB diagnosis. The smear-positive cases for the first sputum were 6.6% (5.98-7.25%), and the second sputum was 6.07% (5.45-6.73%). The Xpert MTB-RIF diagnosis detected 2.93% (10/341) (1.42-5.33%) in the first and 5.44% (16/294) (3.14-8.69%) in the second smear-negative TB samples. The prevalence of Xpert MTB-RIF across smear positive showed that males had 56.87% (178/313) and 56.15% (137/244) and females had 43.13% (135/313) and 43.85% (107/244) for the first and second sputum. Also, false negative smears were 0.18% (10/5607) for smear 1 and 0.31% (16/5126) for smear 2. CONCLUSION: In conclusion, the study highlights the higher sensitivity of the GeneXpert assay compared to traditional smear microscopy for detecting MTB. The GeneXpert assay identified 10 and 16 positive MTB from smear 1 and smear 2 samples which were microscopic negative.

Mosquito bites and stage-specific antibody responses against Plasmodium falciparum in southern Ghana.

BACKGROUND: The human host elicits specific immune responses after exposure to various life stages of the malaria parasite as well as components of mosquito saliva injected into the host during a mosquito bite. This study describes differences in IgG responses against antigens derived from the sporozoite (PfCSP), asexual stage parasite (PfEBA175) and the gametocyte (Pfs230), in addition to an Anopheles gambiae salivary gland antigen (gSG6-P1), in two communities in Ghana with similar blood stage malaria parasite prevalence. METHODS: This study used archived plasma samples collected from an earlier cross-sectional study that enrolled volunteers aged from 6 months to 70 years from Simiw, peri-urban community (N = 347) and Obom, rural community (N = 291). An archived thick and thin blood smear for microscopy was used for the estimation of Plasmodium parasite density and species and DNA extraction from blood spots and P. falciparum confirmation was performed using PCR. This study used the stored plasma samples to determine IgG antibody levels to P. falciparum and Anopheles salivary antigens using indirect ELISA. RESULTS: Individuals from Simiw had significantly higher levels of IgG against mosquito gSG6-P1 [median (95%CI)] [2.590 (2.452-2.783) ng/mL] compared to those from Obom [2.119 (1.957-2.345) ng/mL], p < 0.0001. Both IgG responses against Pfs230proC (p = 0.0006), and PfCSP (p = 0.002) were significantly lower in volunteers from Simiw compared to the participants from Obom. The seroprevalence of PfEBA-175.5R (p = 0.8613), gSG6-P1 (p = 0.0704), PfCSP (p = 0.7798) IgG were all similar in Obom and Simiw. However, Pfs230 seroprevalence was significantly higher at Obom compared to Simiw (p = 0.0006). Spearman correlation analysis showed no significant association between IgG responses against gSG6-P1, PfCSP, Pfs230proC and PfEBA-175.5R and parasite density at both Obom and Simiw (p > 0.05). CONCLUSION: In conclusion, the study showed that participants from Simiw had higher concentrations of circulating gSG6-P1 IgG antibodies but lower concentrations of P. falciparum antibodies, PfCSP IgG and Pfs230proC IgG compared to participants from Obom.

Candidiasis profile at the outpatient department of the university of cape coast hospital in the central region of Ghana: a retrospective study.

INTRODUCTION: Vulvovaginal candidiasis (VVC) is a public health problem with an estimated 138 million women globally experiencing recurrent VVC annually. The microscopic diagnosis of VVC has low sensitivity, but it remains an essential tool for diagnosis as the microbiological culture methods are limited to advanced clinical microbiology laboratories in developing countries. The study retrospectively analyzed the presence of red blood cells (RBCs), epithelial cells (ECs), pus cells (PCs) and Candida albicans positive in wet mount preparation of urine or high vaginal swabs (HVS) samples to test for their sensitivity and specificity for the diagnosis of candidiasis. METHODS: The study is a retrospective analysis at the Outpatient Department of the University of Cape Coast between 2013 and 2020. All urine and high vagina swabs (HVS) cultures samples using Sabourauds dextrose agar with wet mount data were analyzed. 2 × 2 contingency diagnostic test was used to ascertain the diagnostic accuracy of red blood cells (RBCs), epithelial cells (ECs), pus cells (PCs), and Candida albicans positive in wet mount preparation of urine or high vaginal swabs (HVS) samples for the diagnosis of candidiasis. The association of candidiasis among patients' demographics was analyzed using relative risk (RR) analysis. RESULTS: The high prevalence of candida infection was among female subjects 97.1% (831/856) compared to males 2.9% (25/856). The microscopic profiles which characterized candida infection were pus cells 96.4% (825/856), epithelial cells 98.7% (845/856), red blood cells (RBCs) 7.6% (65/856) and Candida albicans positive 63.2% (541/856). There was a lower risk of Candida infections among male patients compared to female patients RR (95% CI) = 0.061 (0.041-0.088). The sensitivity (95%) for detecting Candida albicans positive and red blood cells (0.62 (0.59-0.65)), Candida albicans positive and pus cells (0.75 (0.72-0.78)) and Candida albicans positive and epithelial cells (0.95 (0.92-0.96)) with corresponding specificity (95% CI) of 0.63 (0.60-0.67), 0.69 (0.66-0.72) and 0.74 (0.71-0.76) were detected among the high vaginal swab samples. CONCLUSION: In conclusion, the study has shown that the presence of PCs, ECs, RBCs or ratio of RBCs/ECs and RBCs/PCs in the wet mount preparation from urine or HVS can enhance microscopic diagnosis of VVC cases.

The emergence of chloroquine-sensitive Plasmodium falciparum is influenced by selected communities in some parts of the Central Region of Ghana.

BACKGROUND: The return of chloroquine-sensitive Plasmodium falciparum in sub-Saharan Africa countries offers the opportunity for the reintroduction of chloroquine (CQ) either in combination with other drugs or as a single therapy for the management of malaria. This study assesses the influence of individual study sites on the selection of CQ sensitive P. falciparum markers in the Central region of Ghana. METHODS: Genomic DNA was extracted from an archived filter paper blood blot from Cape Coast, Elmina, Assin Fosu, and Twifo Praso using the Chelex DNA extraction method. The age metadata of the patients from whom the blood spots were taken was collected. The prevalence of CQ-sensitive markers of pfcrt K76 and pfmdr1 N86 was performed using nested PCR and RFLP. The data were analysed using Chi-square and Odd ratio. RESULTS: The overall prevalence of CQ-sensitive P. falciparum markers, pfcrt K76 and pfmdr1 N86 in the Central Region of Ghana were 142 out of 184 (77.17%) and 180 out of 184 (97.83%), respectively. The distribution of pfcrt K76 was assessed among the age groups per the individual study sites. 12 out of 33 (36.36%), 8 out of 33 (24.24%) and 6 out of 33 (18.18%) of pfcrt K76 CQ-sensitive marker were isolated from age 0 to 5 years, 16 to 30 years and 31 to 45 years old respectively at Cape Coast. Assin Fosu and Twifo Praso had the highest pfcrt K76 prevalence in 0-5 years, followed by 16-30 years and 6-15 years of age. The results showed that there was a significant prevalence of pfcrt K76 in all study sites; Cape Coast (χ2 = 26.48, p < 0.0001), Assin Fosu (χ2 = 37.67, p < 0.0001), Twifo Praso (χ2 = 32.25, p < 0.0001) and Elmina (χ2 = 17.88, p < 0.0001). Again, the likelihood to detect pfcrt K76 (OR (95% CI) was 7.105 (3.118-17.14), p < 0.0001 and pfmdr1 (2.028 (1.065-3.790), p < 0.001) among P. falciparum isolates from Cape Coast to be seven times and two times, respectively. CONCLUSION: The study showed a significant selection and expansion of chloroquine-sensitive P. falciparum markers in all the selected study areas in the Central region. This finding has a significant implication for the future treatment, management, and control of P. falciparum malaria.

Antibody responses to two new Lactococcus lactis-produced recombinant Pfs48/45 and Pfs230 proteins increase with age in malaria patients living in the Central Region of Ghana.

BACKGROUND: Recent advances in malaria control efforts have led to an increased number of national malaria control programmes implementing pre-elimination measures and demonstrated the need to develop new tools to track and control malaria transmission. Key to understanding transmission is monitoring the prevalence and immune response against the sexual stages of the parasite, known as gametocytes, which are responsible for transmission. Sexual-stage specific antigens, Pfs230 and Pfs48/45, have been identified and shown to be targets for transmission blocking antibodies, but they have been difficult to produce recombinantly in the absence of a fusion partner. METHODS: Regions of Pfs48/45 and Pfs230 known to contain transmission blocking epitopes, 6C and C0, respectively, were produced in a Lactococcus lactis expression system and used in enzyme linked immunosorbent assays to determine the seroreactivity of 95 malaria patients living in the Central Region of Ghana. RESULTS: Pfs48/45.6C and Pfs230.C0 were successfully produced in L. lactis in the absence of a fusion partner using a simplified purification scheme. Seroprevalence for L. lactis-produced Pfs48/45.6C and Pfs230.C0 in the study population was 74.7 and 72.8%, respectively. CONCLUSIONS: A significant age-dependent increase in antibody titers was observed, which suggests a vaccine targeting these antigens could be boosted during a natural infection in the field.

Use of proscribed chloroquine is associated with an increased risk of pfcrt T76 mutation in some parts of Ghana.

BACKGROUND: After years of disuse of chloroquine (CQ) as first-line anti-malarial drug in Ghana, reports from molecular studies conducted in parts of the country indicate varying prevalence of T76 mutation in the pfcrt gene. This situation has several health implications, one being that mutations that confer resistance to CQ have been reported to show substantial cross-resistance to other anti-malarial drugs. It is important to identify some of the factors contributing to the continuous presence of CQ resistance markers in the country. This study determined the prevalence of T76 mutation in pfcrt gene of Plasmodium falciparum isolates collected from selected areas of the Central region of Ghana and correlated with the level of CQ use in these areas. METHODS: Plasmodium falciparum DNA was extracted from collected blood-blot filter paper samples in the study sites. The prevalence of T76 point mutation in pfcrt gene was assessed using nested PCR followed by RFLP. CQ from pharmacy and chemical shops was obtained using mystery buying method. The extent of CQ use by the participants was determined by measuring the level of the drug in their urine samples using the Saker-Solomon method. RESULTS: Of the 214 P. falciparum isolates analysed, 71.9% were found to have T76 mutation of pfcrt gene. The study revealed that 14.49% of community pharmacies and chemical shops had stocks of CQ for sale while 16.9% of the participants had CQ in their urine samples. There is five times more risks of becoming infected with CQ resistant strain for staying in an area where CQ is stocked for sale [RR = 0.20, p < 0.0001] and thirteen times more risks of having CQ-resistant mutant from those who still use CQ than non-users [OR = 0.08, p < 0.0001]. CONCLUSION: This study has shown that high variation in the prevalence of T76 mutations of P. falciparum is linked with the level of CQ stocking and usage within study area.

Care practices of contact lens solutions and microbial contamination among wearers in Ghana.

PURPOSE: This study sought to assess contact lens solutions care practices, and their microbial contamination among contact lens wearers in Ghana and to profile their antibiotic susceptibility pattern. METHODS: The study employed a biphasic approach which involved a cross-sectional design that investigated participants' habits related to care for the solutions with a two-part questionnaire and a microbiological analysis of samples of contact lens care solutions of the participants for microbial contamination. A snowball sampling method provided access to 32 different contact lens wearers in four care facilities in Ghana. In most cases, the participants had no pre-existing familial relationship with each other or with the care facilities. RESULTS: Out of 32 samples of contact lens solutions, 30 were tested for microbial contamination. A total of 23 (76.67 %) samples of contact lens solution were found to be contaminated with Enterobacter sp. (34.80 %), Pseudomonas sp. (21.70 %), Bacilli sp. (21.70 %), Klebsiella sp. (17.20 %), and Escherichia coli (4.60 %). The duration of solution storage in the open bottle and nonadherence to manufacturer instructions for solution storage showed a statistically significant association with microbial contamination (p ≤ 0.05). CONCLUSION: Contact lens care solutions have been found to harbour multiple antibiotic-resistant bacteria that are potentially pathogenic to the corneal surface. The contamination is associated with some unhealthy solution-care practices among wearers.

Correction: Genotypic glucose-6-phosphate dehydrogenase (G6PD) deficiency protects against Plasmodium falciparum infection in individuals living in Ghana.

[This corrects the article DOI: 10.1371/journal.pone.0257562.].

Plant diversity and ethnobotanical importance of home gardens in Ghana's middle belt: a cross-sectional survey of the Sunyani municipality.

BACKGROUND: Home gardens are a species-rich socioecological system with a diverse range of cultivated and naturally occurring plants with the potential to make contributions to address sustainable food, biodiversity and climate crisis. However, there is a dearth of information on the socio-demographic profile of home gardeners and the importance of home gardens to ethnobotany, food security and biodiversity. Therefore, the study aimed to assess the socio-demographic profile of home gardeners in the Sunyani municipality as a case in point for the middle belt of Ghana and to evaluate the diversity of plants in home gardens and their ethnobotanical importance. METHODS: A total of 12 suburbs were selected from three subzones in the Sunyani municipality. In each suburb, 25% of households were randomly selected and if they had a home garden, one adult in the house was interviewed. A list of all plants in the home garden and their uses was obtained from respondents. A Chi-square test was used to assess the distribution of home gardeners among various socio-demographic categories, and binomial logistic regression was employed to determine links between socio-demography and home garden attributes. The ethnobotany R package was used to evaluate the ethnobotanical importance of plants in home gardens. RESULTS: A total of 186 respondents were recruited for this study, 79 being females and 107 males. A total of 79 plant species were also identified belonging to 70 genera and 40 families. Trees were the most common plant life form in home gardens, followed by shrubs, herbs, vines, grasses and lianas. Ethnobotanical indices revealed the most important plants in home gardens to be staples, food supplements and medicinal plants. These were Musa paradisiaca, Caripa pabaya, Xanthosoma sagittifolium, Manihot utilisima and Mangifera indica, Moringa oleifera, Citrus sinensis, Capsicum frutescens, Taraxacum officinale, Solanum aethiopicum, Cocos nucifera, Solanum torvum, Persea americana, Dioscorea alata and Elaeis guineensis. CONCLUSION: Plants used as staples, food supplements and medicinal purposes emerged as the most culturally relevant scoring high on all ethnobotanical indices. Home gardens present an opportunity to address food security and nutrition needs of households and communities.

Abnormal Obesity Phenotype Is Associated with Reduced eGFR among Diabetes Mellitus and Hypertensive Patients in a Peri-Urban Community in Ghana.

Background: Diabetes mellitus (DM) is a chronic disease characterized by hyperglycemia due to obesity and defects in insulin action. Significant complications of DM include kidney disease due to its association with hypertension and obesity. Thus, the contribution of the various obesity phenotypes to the kidney impairment observed among hypertensive and diabetes mellitus patients is of major concern. Aim: The study assessed the association between obesity phenotypes and reduced glomerular filtration rate among diabetes mellitus and hypertensive patients. Methods: Three hundred and ten (310) adult patients diagnosed with type 2 diabetes mellitus, hypertension, or both who attended the Presbyterian Hospital, Dormaa Ahenkro, from October 2016 to March 2017 were recruited for the study. Blood samples were collected to analyze biochemical parameters (fasting blood glucose (FBG), lipid profile, and creatinine). Questionnaires were used to collect sociodemographic information, and anthropometrics were appropriately measured. The estimated glomerular filtration rate (eGFR) was calculated using the CKD-EPI equation, and reduced eGFR was defined as eGFR <90 ml/min/1.73 m2. Results: The prevalence of metabolically healthy nonobese (MHNO), metabolically healthy obese (MHO), metabolically abnormal nonobese (MANO), and metabolically abnormal obese (MAO) phenotypes among the study participants was 30.65%, 4.50%, 52.90%, and 11.94%, respectively. The highest prevalence of reduced eGFR (29/37 (78.38%)) was seen among the MAO group. This was followed by the MANO, MHO, and MHNO with a reduced eGFR prevalence of 62.20%, 57.64%, and 37.89%, respectively. After normalization with MHNO, the reduced eGFR was 1.51, 1.64, and 2.06 times expressed in MHO, MANO, and MAO. For the total samples, when MHNO was maintained as a reference, reduced eGFR was significantly associated with MANO (aOR = 3.07 (95% CI = 1.76-5.35), P < 0.001) and MAO (aOR = 5.67 (95% CI = 2.66-17.27), P < 0.001) even after adjusting for age, gender, smoking, and alcohol intake. This association was maintained among the female study participants when stratified by gender, and in addition, among the female participants, reduced eGFR was also associated with MHO (aOR = 4.19 (95% CI = 1.06-16.53), P=0.041). Conclusion: There is a high prevalence of abnormal metabolic phenotypes among diabetes mellitus patients, and these were significantly associated with reduced eGFR among our study participants.

Intestinal microbiome-rheumatoid arthritis crosstalk: The therapeutic role of probiotics.

Rheumatoid arthritis (RA) is a common systemic autoimmune disease with a global health importance. It is characterized by long-term complications, progressive disability and high mortality tied to increased social-economic pressures. RA has an inflammatory microenvironment as one of the major underlying factors together with other complex processes. Although mechanisms underlying the triggering of RA remain partially elusive, microbiota interactions have been implicated. Again, significant alterations in the gut microbiome of RA patients compared to healthy individuals have intimated a chronic inflammatory response due to gut dysbiosis. Against this backdrop, myriads of studies have hinted at the prospective therapeutic role of probiotics as an adjuvant for the management of RA in the quest to correct this dysbiosis. In this article, the major gut microbiome alterations associated with RA are discussed. Subsequently, the role of the gut microbiome dysbiosis in the initiation and progression of RA is highlighted. Lastly, the effect and mechanism of action of probiotics in the amelioration of symptoms and severity of RA are also espoused. Although strain-specific, probiotic supplementation as adjuvant therapy for the management of RA is very promising and warrants more research.

In vitro regeneration of Vitellaria paradoxa from shoot tip explants.

Shea tree (Vitellaria paradoxa) is an essential tree crop with great potential economic value mainly because of its seed oil (shea butter) which is of high demand for manufacturing assorted products in food, cosmetic, and rubber industries. Propagation of this species is, however, hindered by relative unavailability of seed (nuts), erratic seed germination, a long vegetative phase, and latex exudation from cuttings. Thus, another method of propagation through in vitro culture is recommended for rapid multiplication of shea genotypes for large-scale cultivation. In the present study, the effects of two cytokinins, namely, 6-benzylaminopurine (BAP) and kinetin (KIN), and one auxin, namely 1-naphthaleneacetic acid (NAA), on shoot and/or root induction in vitro were assessed at various combinations/concentrations. The inclusion of these growth regulators in the culture medium significantly improved (P < 0.05) shoot/root regeneration over the controls. The highest shoot regeneration percentage (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 2 mg ∙ dm-3 KIN + + 0.5 mg ∙ dm-3 NAA or 1.5 mg ∙ dm-3 KIN within 7/8 days of inoculation. This medium (2 mg ∙ dm-3 KIN + + 0.5 mg ∙ dm-3 NAA) showed the highest mean shoot length of 3.24 cm. Compared to KIN, BAP was more effective in inducing vigorous shoot growth. However, rooting was induced only on MS medium modified with 1 mg ∙ dm-3 BAP + 0.5 mg ∙ dm-3 NAA. These findings can serve as baseline information for in vitro, commercialscale propagation of shea tree.

Evaluating immunohaematological profile among COVID-19 active infection and recovered patients in Ghana.

INTRODUCTION: The rapid spread of COVID-19 has been a global public health problem and it is yet to be put under control. Active COVID-19 is associated with unrestrained secretion of pro-inflammatory cytokines and imbalances in haematological profile including anaemia, leukocytosis and thrombocytopaenia. However, the haematological profile and immune status following recovery from COVID-19 has not been recognized. We evaluated the immunohaematological profile among COVID-19 patients with active infection, recovered cases and unexposed healthy individuals in the Ashanti region of Ghana. METHODOLOGY: A total of 95 adult participants, consisting of 35 positive, 30 recovered and 30 unexposed COVID-19 negative individuals confirmed by RT-PCR were recruited for the study. All the patients had the complete blood count performed using the haematological analyzer Sysmex XN-1500. Their plasma cytokine levels of interleukin (IL)-1ß, IL-6, IL-10, IL-17, tumour necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) were analysed using ELISA. Statistical analyses were performed on R statistical software. RESULT: The Patients with COVID-19 active infection had significantly higher levels of IL10 (181±6.14 pg/mL vs 155.00±14.32 pg/mL vs 158.80±11.70 pg/mL, p = 0.038), WBC count (5.5±0.4 x109 /L vs 4.5±0.6 x109 /L vs 3.8±0.5, p < 0.0001) and percentage basophil (1.8±0.1% vs 0.8±0.3% vs 0.7±0.2%, p = 0.0040) but significantly lower levels of IFN-γ (110.10±9.52 pg/mL vs 142.80±5.46 pg/mL vs 140.80±6.39 pg/mL, p = 0.021), haematocrit (24.1±3.7% vs 38.3± 3.0% vs 38.5±2.2%, p < 0.0001), haemoglobin concentration (9.4±0.1g/dl vs 12.5± 5.0g/dl vs 12.7±0.8, p < 0.0001) and MPV (9.8±0.2fL vs 11.1±0.5fL vs 11.6±0.3fL, p < 0.0001) compared to recovered and unexposed controls respectively. There were significant association between IL-1ß & neutrophils (r = 0.42, p<0.05), IL-10 & WBC (r = 0.39, p<0.05), IL-10 & Basophils (r = -0.51, p<0.01), IL-17 & Neutrophil (r = 0.39, p<0.05) in the active COVID-19 cases. CONCLUSION: COVID-19 active infection is associated with increased IL-10 and WBC with a concomitant decrease in IFN-γ and haemoglobin concentration. However, recovery from the disease is associated with immune recovery with appareantly normal haematological profile.

Nationwide Surveillance of Pfhrp2 Exon 2 Diversity in Plasmodium falciparum Circulating in Symptomatic Malaria Patients Living in Ghana.

Reports of increasing false-negative HRP2-based rapid diagnostic test results across Africa require constant monitoring of factors associated with these false-negative outcomes, as failure of this diagnostic tool will have severe consequences on malaria treatment and control programs. This study characterized the extent of genetic diversity in the Plasmodium falciparum histidine-rich protein 2 (Pfhrp2) gene in P. falciparum isolates from symptomatic malaria patients across the regions of Ghana. Exon 2 of Pfhrp2 was amplified from gDNA using polymerase chain reaction. All Pfhrp2-negative samples were subjected to Pf18S rRNA and Pfmsp2 gene amplifications. The amplified Pfhrp2 exon 2 fragments from clonal samples were sent for commercial Sanger sequencing. The type and number of PfHRP2 repeats, classified based on repeat types previously reported, were estimated from the sequence data and compared among geographical regions. About 81% (2,333/2,890) of the original microscopy positive DBS were available and used in this study. The Pfhrp2 exon 2 amplification was successful in 98.5% (2,297/2,333) of the tested samples, with band size ranging from 400 bp to 1,050 bp. A total of 13 out of the 24 previously reported repeat types were identified among the samples, with three samples lacking both type 2 and type 7 repeat motifs. This study suggested that the genetic diversity of Pfhrp2 exon 2 identified in P. falciparum circulating in symptomatic malaria patients in Ghana is unlikely to influence the sensitivity and specificity of HRP2 RDT-based diagnosis.

Antibiotic-resistant pathogenic bacterial isolates from patients attending the outpatient department of university of Cape Coast hospital, Ghana: A retrospective study between 2013-2015.

Uropathogenic Escherichia coli (E. coli) is an important urinary tract infection (UTI) that has been associated with both complicated and uncomplicated disease conditions. The global emergence of multiple drug-resistant (MDR) and extended-spectrum ß-lactamase (ESBL) is of public health concern as the resistance limits the current treatment options. The objective of this study was to analyze the antibiotic-resistant patterns among the uropathogenic E. coli isolates at the University of Cape Coast (UCC) hospital between 2013 and 2015 as baseline data to understand the current antibiotic resistance situation within UCC and its environs. A retrospective cross-sectional study of bacteria isolates at UCC hospital from January 2013 to December 2015 were analyzed. A standard biochemical and antibiotic susceptibility tests were performed using Kirby-Bauer NCCLs modified disc diffusion technique. The network of interaction between pathogenic isolates and antibiotic resistance was performed using Cytoscape software. Statistical significance was tested using ANOVA and one-sample Wilcoxon test. The overall E. coli prevalence was 15.76% (32/203); females had the highest infection of 17.33% (26/150) compared to male subjects who had 11.32% (6/53) out of all the pathogenic infections. The E. coli prevalence among the age categories were 2/21 (9.52%), 27/154 (17.53%) and 4/21 (19.05%) among ≤20 years, 21-40 years and 41-60 years respectively. The isolated resistant pathogens exhibited different antibiotic resistance patterns. An interaction network of nodes connecting to other nodes indicating positive correlations between the pathogens and antibiotic resistance was established. Escherichia coli, Citrobacter spp, Klebsiella spp among other isolated pathogens formed higher centrality in the network of interaction with antibiotic resistance. The individual E. coli isolates showed a significant difference in the mean ± SD (95% CI) pattern of antibiotic resistance, 2.409±1.205 (1.828-2.990), χ2 = 36.68, p<0.0001. In conclusion, the study reports the interaction of E. coli isolates at UCC hospital and its antibiotic-resistant status between 2013 and 2015. This data forms the baseline information for assessing the current antibiotic status in UCC and its environs.

Nationwide molecular surveillance of three Plasmodium species harboured by symptomatic malaria patients living in Ghana.

BACKGROUND: Clinical presentations of malaria in Ghana are primarily caused by infections containing microscopic densities of Plasmodium falciparum, with a minor contribution from Plasmodium malariae and Plasmodium ovale. However, infections containing submicroscopic parasite densities can result in clinical disease. In this study, we used PCR to determine the prevalence of three human malaria parasite species harboured by suspected malaria patients attending healthcare facilities across the country. METHODS: Archived dried blood spots on filter paper that had been prepared from whole blood collected from 5260 patients with suspected malaria attending healthcare facilities across the country in 2018 were used as experimental material. Plasmodium species-specific PCR was performed on DNA extracted from the dried blood spots. Demographic data and microscopy data for the subset of samples tested were available from the original study on these specimens. RESULTS: The overall frequency of P. falciparum, P. malariae and P. ovale detected by PCR was 74.9, 1.4 and 0.9%, respectively. Of the suspected symptomatic P. falciparum malaria cases, 33.5% contained submicroscopic densities of parasites. For all regions, molecular diagnosis of P. falciparum, P. malariae and P. ovale was significantly higher than diagnosis using microscopy: up to 98.7% (75/76) of P. malariae and 97.8% (45/46) of P. ovale infections detected by PCR were missed by microscopy. CONCLUSION: Plasmodium malariae and P. ovale contributed to clinical malaria infections, with children aged between 5 and 15 years harbouring a higher frequency of P. falciparum and P. ovale, whilst P. malariae was more predominant in individuals aged between 10 and 20 years. More sensitive point-of-care tools are needed to detect the presence of low-density (submicroscopic) Plasmodium infections, which may be responsible for symptomatic infections.

Genotypic glucose-6-phosphate dehydrogenase (G6PD) deficiency protects against Plasmodium falciparum infection in individuals living in Ghana.

INTRODUCTION: The global effort to eradicate malaria requires a drastic measure to terminate relapse from hypnozoites as well as transmission via gametocytes in malaria-endemic areas. Primaquine has been recommended for the treatment of P. falciparum gametocytes and P. vivax hypnozoites, however, its implementation is challenged by the high prevalence of G6PD deficient (G6PDd) genotypes in malaria endemic countries. The objective of this study was to profile G6PDd genotypic variants and correlate them with malaria prevalence in Ghana. METHODS: A cross-sectional survey of G6PDd genotypic variants was conducted amongst suspected malaria patients attending health care facilities across the entire country. Malaria was diagnosed using microscopy whilst G6PD deficiency was determined using restriction fragment length polymorphisms at position 376 and 202 of the G6PD gene. The results were analysed using GraphPad prism. RESULTS: A total of 6108 subjects were enrolled in the study with females representing 65.59% of the population. The overall prevalence of malaria was 36.31%, with malaria prevalence among G6PDd genotypic variants were 0.07% for A-A- homozygous deficient females, 1.31% and 3.03% for AA- and BA- heterozygous deficient females respectively and 2.03% for A- hemizygous deficient males. The odd ratio (OR) for detecting P. falciparum malaria infection in the A-A- genotypic variant was 0.0784 (95% CI: 0.0265-0.2319, p<0.0001). Also, P. malariae and P. ovale parasites frequently were observed in G6PD B variants relative to G6PD A- variants. CONCLUSION: G6PDd genotypic variants, A-A-, AA- and A- protect against P. falciparum, P. ovale and P. malariae infection in Ghana.

Mycophenolate-induced oral ulcers: Case report and literature review.

PURPOSE: A case of mycophenolate mofetil (MMF)-induced oral ulceration in a kidney transplant recipient is reported. SUMMARY: A 54-year-old man who had received a kidney transplant 7 months previously reported to our outpatient clinic with severe oral ulcers with odynophagia and was admitted to the hospital. His maintenance immunosuppressive agents at the time of admission consisted of tacrolimus and mycophenolate. The patient had stable renal function, with all laboratory values within normal ranges. After various alternative etiologies were ruled out, drug-induced oral ulceration was suspected, and the patient's tacrolimus dose was empirically reduced, resulting in reduction of the trough concentration from 10 ng/mL to 3.3 ng/mL without improvement of the ulceration. Mycophenolate-induced oral ulceration was suspected, and MMF was discontinued. Within 5 days of discontinuation, there was a remarkable improvement in both the size and severity of the ulceration, and the patient was discharged from the hospital. During the next clinic visit (a total of 12 days after MMF was discontinued), the patient's mouth and esophageal ulcers had completely healed. Six weeks after complete resolution of the ulcer, MMF at a dosage of 250 mg twice daily was initiated; the dosage was subsequently increased to 500 mg twice daily without a recurrence of ulceration. CONCLUSION: A 54-year-old man developed oral ulceration after 7 months of MMF therapy. Discontinuation of therapy resulted in prompt resolution of the patient's ulcers, with no recurrence of ulceration at a lower MMF dose. This is the first case report indicating that mycophenolate-induced ulceration may be dose dependent.

Fibroblast Growth Factor-21 ameliorates hepatic encephalopathy by activating the STAT3-SOCS3 pathway to inhibit activated hepatic stellate cells.

Neurological dysfunction, one of the consequences of acute liver failure (ALF), and also referred to as hepatic encephalopathy (HE), contributes to mortality posing challenges for clinical management. FGF21 has been implicated in the inhibition of cognitive decline and fibrogenesis. However, the effects of FGF21 on the clinical and molecular presentations of HE has not been elucidated. HE was induced by fulminant hepatic failure using thioacetamide (TAA) in male C57BL/6J mice while controls were injected with saline. For two consecutive weeks, mice were treated intraperitoneally with FGF21 (3 mg/kg) while controls were treated with saline. Cognitive, neurological, and activity function scores were recorded. Serum, liver, and brain samples were taken for analysis of CCL5 and GABA by ELISA, and RT qPCR was used to measure the expressions of fibrotic and pro-inflammatory markers. We report significant improvement in both cognitive and neurological scores by FGF21 treatment after impairment by TAA. GABA and CCL5, key factors in the progression of HE were also significantly reduced in the treatment group. Furthermore, the expression of fibrotic markers such as TGFß and Col1 were also significantly downregulated after FGF21 treatment. TNFα and IL-6 were significantly reduced in the liver while in the brain, TNFα and IL-1 were downregulated. However, both in the liver and the brain, IL-10 was significantly upregulated. FGF21 inhibits CXCR4/CCL5 activation and upregulates the production of IL-10 in the damaged liver stimulating the production pro-inflammatory cytokines and apoptosis of hepatic stellate cells through the STAT3-SOCS3 pathway terminating the underlying fibrosis in HE.