Immunological and biochemical biomarker alterations among SARS-COV-2 patients with varying disease phenotypes in Uganda.

Every novel infection requires an assessment of the host response coupled with identification of unique biomarkers for predicting disease pathogenesis, treatment targets and diagnostic utility. Studies have exposed dysregulated inflammatory response induced by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as significant predictor or cause of disease severity/prognosis and death. This study evaluated inflammatory biomarkers induced by SARS-CoV-2 in plasma of patients with varying disease phenotypes and healthy controls with prognostic or therapeutic potential. We stratified SARS-CoV-2 plasma samples based on disease status (asymptomatic, mild, severe, and healthy controls), as diagnosed by RT-PCR SARS-CoV-2. We used a solid phase sandwich and competitive Enzyme-Linked Immunosorbent Assay (ELISA) to measure levels of panels of immunological (IFN-γ, TNF-α, IL-6, and IL-10) and biochemical markers (Ferritin, Procalcitonin, C-Reactive Protein, Angiotensin II, Homocysteine, and D-dimer). Biomarker levels were compared across SARS-CoV-2 disease stratification. Plasma IFN-γ, TNF-α, IL-6, and IL-10 levels were significantly (P < 0.05) elevated in the severe SARS-CoV-2 patients as compared to mild, asymptomatic, and healthy controls. Ferritin, Homocysteine, and D-dimer plasma levels were significantly elevated in severe cases over asymptomatic and healthy controls. Plasma C-reactive protein and Angiotensin II levels were significantly (P < 0.05) higher in mild than severe cases and healthy controls. Plasma Procalcitonin levels were significantly higher in asymptomatic than in mild, severe cases and healthy controls. Our study demonstrates the role of host inflammatory biomarkers in modulating the pathogenesis of COVID-19. The study proposes a number of potential biomarkers that could be explored as SARS-CoV-2 treatment targets and possible prognostic predictors for a severe outcome. The comprehensive analysis of prognostic biomarkers may contribute to the evidence-based management of COVID-19 patients.

Retrospective in silico mutation profiling of SARS-CoV-2 structural proteins circulating in Uganda by July 2021: Towards refinement of COVID-19 disease vaccines, diagnostics, and therapeutics.

The SARS-CoV-2 virus, the agent of COVID-19, caused unprecedented loss of lives and economic decline worldwide. Although the introduction of public health measures, vaccines, diagnostics, and therapeutics disrupted the spread of the SARS-CoV-2, the emergence of variants poses substantial threat. This study traced SARS-CoV-2 variants circulating in Uganda by July 2021 to inform the necessity for refinement of the intervention medical products. A comprehensive in silico analysis of the SARS-CoV-2 genomes detected in clinical samples collected from COVID-19 patients in Uganda revealed occurrence of structural protein variants with potential of escaping detection, resisting antibody therapy, or increased infectivity. The genome sequence dataset was retrieved from the GISAID database and the open reading frame encoding the spike, envelope, membrane, or nucleocapsid proteins was translated. The obtained protein sequences were aligned and inspected for existence of variants. The variant positions on each of the four alignment sets were mapped on predicted epitopes as well as the 3D structures. Additionally, sequences within each of the sets were clustered by family. A phylogenetic tree was constructed to assess relationship between the encountered spike protein sequences and Wuhan-Hu-1 wild-type, or the Alpha, Beta, Delta and Gamma variants of concern. Strikingly, the frequency of each of the spike protein point mutations F157L/Del, D614G and P681H/R was over 50%. The furin and the transmembrane serine protease 2 cleavage sites were unaffected by mutation. Whereas the Delta dominated the spike sequences (16.5%, 91/550), Gamma was not detected. The envelope protein was the most conserved with 96.3% (525/545) sequences being wild-type followed by membrane at 68.4% (397/580). Although the nucleocapsid protein sequences varied, the variant residue positions were less concentrated at the RNA binding domains. The dominant nucleocapsid sequence variant was S202N (34.5%, 205/595). These findings offer baseline information required for refining the existing COVID-19 vaccines, diagnostics, and therapeutics.

Molecular epidemiology of anaplasmosis in small ruminants along a human-livestock-wildlife interface in Uganda.

BACKGROUND: Information as regards the epidemiology of the Anaplasmataceae in small ruminants in several low- and middle-income countries is scarce. METHODS: In this study a total of 712 DNA samples collected from small ruminants were analyzed for Anaplasmataceae and Anaplasma ovis using the 16S rRNA and MSP4 genes respectively. Infection risk was assessed by location, sex and age of the animals and qGIS® was used to construct spatial maps. RESULTS: The prevalence of Anaplasmataceae spp was 89.1% (95% CI: 77.5-95.9) and 79.1% (95% CI: 75.9-82.1) in ovines and caprines respectively (RR = 1.1, 95% CI: 1.0-1.3); higher than those previously reported in other eastern African countries. The prevalence of A. ovis was 26.1% and 25.4% for both ovines and caprines respectively with ovines showing significantly higher levels of infection than caprines (P < 0.05). The risk of Anaplasma ovis infections was not affected by age (OR = 1.2, 95% CI: 0.9-1.7) or sex (OR = 1.1, 95% CI: 0.6-2.0). Small ruminants located at the forest edge (<0.3 km) showed higher A. ovis prevalence than those found inland with infections present in the midland regions associated with increased agricultural activity. CONCLUSION: Anaplasma ovis remains a major challenge for small ruminant husbandry in Uganda and infections are under-reported. Policy efforts to prioritize management of Anaplasmataceae for small ruminant health would promote livestock productivity in vulnerable communities, improving livelihoods and ecosystem health.

Prevalence of hemoprotozoan parasites in small ruminants along a human-livestock-wildlife interface in western Uganda.

Small ruminants are important to community livelihood in developing countries; however information on the role of hemoprotozoan parasites is scanty. The objective of the study was to determine hemoprotozoan parasitic prevalence in western Uganda and identify major areas associated with these infections. This was a cross sectional study conducted at the edge of Budongo Conservation Forest in Masindi district of western Uganda in which 712 small ruminants were sampled. Blood from the jugular vein was collected from caprines and ovines and placed in an EDTA tube, and transported to the laboratory for examination. Thin and thick smears were prepared and examined by microscopy for hemoprotozoan parasites, and DNA was extracted and examined by PCR for Trypanosoma spp. A total of 13 villages in Budongo sub-county were surveyed and the study showed that caprines were the major small ruminants of importance to the community. Prevalence of hemoprotozoan parasites was as follows; anaplasmosis (3.65%) > theileriosis (0.45%) > trypanosomiasis (0.15%) and babesiosis (0%) by microscopy. Infections were found in the young with the exception of Anaplasma spp. while coinfections of anaplasmosis and theileriosis were high. Molecular analysis showed an overall trypanosome prevalence of 9.27% (PCR), mainly due to Trypanosoma brucei and T. congolense forest. Villages with trypanosomiasis were found in lowlands and swamps. The current trypanosomiasis prevalence in small ruminants of Uganda was 10 times greater than that previously reported showing that the disease burden has increased overtime within Uganda. A prevalence of 0.14% (95% CI: 0.00, 0.78) for the SRA gene showed that small ruminants would be important reservoirs of infection to humans. Hemoprotozoan parasites are a threat to community livelihood in developing countries and the role of molecular diagnostic techniques in disease monitoring was re-emphasized by this study. Information on primary hosts involved in the propagation of hemoprotozoan parasites in Uganda would help streamline prospective disease surveillance and control efforts.

Grain Amaranth Is Associated with Improved Hepatic and Renal Calcium Metabolism in Type 2 Diabetes Mellitus of Male Wistar Rats.

BACKGROUND: Dysregulation of calcium signaling is a hallmark of diabetes mellitus (DM) and grain amaranth (AG) has antidiabetic properties. Information on the mechanism of action of AG on blood, renal, and hepatic tissues is sparse, although it continues to be an important alternative medicinal plant in several developing countries. The objective of the study was to determine key changes in calcium levels and s100a1 protein levels and antioxidant and histopathologic changes in blood, renal, and hepatic tissues of male diabetic Wistar rats. MATERIALS AND METHODS: This was an experimental study in which 30 male Wistar rats were kept for 5 weeks (6 groups, N =5). Groups 1-IV had T2DM induced using Nicotinamide and Streptozotocin: Group I, Mixtard®; group II, positive control; group III, 25% AG; group IV, 50% AG. Furthermore, group V consisted of normal rats given 50% GA and group VI was negative control. Blood, renal, and hepatic tissues were collected and analyzed for calcium, s100a1 protein levels, and antioxidant and histopathological changes. RESULTS AND DISCUSSION: In blood, renal, and hepatic tissue, calcium and s100a1 levels were low during T2DM and these increased following AG supplementation. This was important for improved metabolic processes, thus leading to the low malondialdehyde (MDA) and glutathione peroxidase (GPx) activity in the tissues. Efficient antioxidant status was important for improved calcium signaling mechanisms, thus leading to improved tissue function and protection demonstrating the importance of AG as an alternative medicinal source through the calcium signaling pathway. CONCLUSION: Grain amaranth exerts its antidiabetic properties through improved calcium homeostasis in blood, kidney, and liver.

Trypanosoma brucei brucei traverses different biological barriers differently and may modify the host plasma membrane in the process.

Trypanosoma brucei are extracellular hemoflagellate protozoan parasites and one of the causative agents of a devastating zoonotic disease called African Trypanosomiasis. In humans, the disease is caused by Trypanosoma brucei rhodensiense and Trypanosoma brucei gambiense, which cross the blood brain barrier (BBB) causing neurological disorders which culminate in death if untreated. In some domestic animals and laboratory rodents, Trypanosoma brucei brucei causes a disease similar to that in humans. The mechanism by which Trypanosoma brucei brucei invade biological barriers including the BBB has not been fully elucidated. To further address this issue, Mardin Dardy Canine Kidney II (MDCKII) and Human dermal microvascular endothelial cell (HDMEC) monolayers were grown to confluence on transwell inserts to constitute in vitro biological barriers. MDCKII cells were chosen for their ability to form tight junctions similar to those formed by the BBB endothelial cells. Labeled trypanosomes were placed in the upper chamber of transwell inserts layered with confluent MDCKII/HDMEC monolayers and their ability to cross the monolayer over time evaluated. Our results show that only 0.5-1.25% of Trypanosoma brucei brucei were able to migrate across the monolayers after 3 h. By employing immune-staining and confocal microscopic analysis we observed that trypanosomes were located at the tight junctions and inside the cell in the MDCK II monolayers indicating that they crossed the monolayer using both the paracellular and transcellular routes. Our observations also showed that there seemed to be no obvious degradation of junction proteins Zonula Ocludens-1, Occludin and Ecadherin. In the HDMEC cell monolayer, our scanning electron microscopy data showed that Trypanosoma brucei brucei is able to modulate the plasma membrane to form invaginations similar to cuplike structures formed by Tlymphocytes. However these structures seemed to be independent of vascular adhesion molecules suggesting that they could be more like the membrane ruffles formed by certain intracellular bacteria during invasion. Taken together, our data reveal a mechanism by which Trypanosoma brucei brucei is able to cross different biological barriers including the BBB without causing any obvious damage.