Peroxynitrite-mediated inactivation of heme oxygenases

BACKGROUND: Endogenous nitric oxide (NO) and carbon monoxide (CO) are generated by nitric oxide synthase and heme oxygenase, respectively. Like NO, CO has been accepted as an important cellular signaling molecule in biological systems. An up-regulation in both gene and protein expression of heme oxygenase-1 (HO-1) under oxidative/nitrosative stress has been well documented, and the protective role of HO-1 and HO-2 against oxidative damage is proposed. However, data on the direct effect of reactive oxygen/nitrogen species (ROS/RNS) on HO function is incomplete. Using gas chromatography to quantify carbon monoxide (CO) formation from heme oxidation, we investigated the effects of peroxynitrite (ONOO(-)) on the in vitro catalytic activity of rat spleen (HO-1) and brain (HO-2) microsomal heme oxygenases. RESULTS: Exposure to ONOO(- )led to concentration-dependent but reversible decreases in the activity of microsomal rat spleen and brain HO activity. Spleen HO activity was 100-fold more sensitive to ONOO(-)-dependent inactivation compared to that of the brain, with IC(50 )values of 0.015 ± 0.005 mM and 1.25 ± 0.25 mM respectively. Inhibition of both rat spleen and brain microsomal HO activity was also observed with tetra-nitromethane, a tyrosine nitrating agent, as well as two NO donors, S-nitrosoglutathione (GSNO) and diethylamine NONOate (DEA-NONOate). However, no additive effect was found following the application of NO donors and ONOO(- )together. CONCLUSION: These results indicate that ONOO(- )may regulate HO-1 and HO-2 activities by mechanisms that involve different interactions with these proteins. It is suggested that while nitration of tyrosine residues and oxidation of sulfhydryl groups may be involved, consideration should be given to other facets of ONOO(- )chemistry. This inhibition of HO activity offers a mechanism for cross talk between the nitric oxide synthase and HO systems

A systematic review of experimental evidence for antiviral effects of ivermectin and an in silico analysis of ivermectin's possible mode of action against SARS-CoV-2

Viral infections remain a major cause of economic loss with an unmet need for novel therapeutic agents. Ivermectin is a putative antiviral compound; the proposed mechanism is the inhibition of nuclear translocation of viral proteins, facilitated by mammalian host importins, a necessary process for propagation of infections. We systematically reviewed the evidence for the applicability of ivermectin against viral infections including SARS-CoV-2 regarding efficacy, mechanisms and selective toxicity. The SARS-CoV-2 genome was mined to determine potential nuclear location signals for ivermectin and meta-analyses for in vivo studies included all comparators over time, dose range and viral replication in multiple organs. Ivermectin inhibited the replication of many viruses including those in Flaviviridae, Circoviridae and Coronaviridae families in vitro. Real and mock nuclear location signals were identified in SARS-CoV-2, a potential target for ivermectin and predicting a sequestration bait for importin β, stopping infected cells from reaching a virus-resistant state. While pharmacokinetic evaluations indicate that ivermectin could be toxic if applied based on in vitro studies, inhibition of viral replication in vivo was shown for Porcine circovirus in piglets and Suid herpesvirus in mice. Overall standardized mean differences; 95% confidence intervals for ivermectin versus controls were: -4.43 (-5.81, -3.04), P < 0.00001. Based on current results, the potential for repurposing ivermectin as an antiviral agent is promising. However, further work is needed to reconcile in vitro studies with clinical efficacy. Developing ivermectin as an additional antiviral agent should be pursued with an emphasis on pre-clinical trials in validated models of infection

Utilisation and evaluation of cooperative case-based teaching for integration of microbiology and pharmacology in veterinary education

Purpose: Integrating basic sciences with clinical disciplines while fostering clinical reasoning capabilities is difficult. We investigated the utilisation of diagnostic specimens and, a cooperative, case-based learning and teaching model to integrate principles of antimicrobial drug pharmacology and microbiology in the fifth year of a veterinary course. Methods: In small groups, students were assigned diagnostic specimens from which they isolated and identified clinically relevant microorganisms and then performed antimicrobial susceptibility tests based on a review of pharmacology, microbiology and pathophysiology. Results were recorded and analysed followed by a student-led integrative tutorial. Learning outcomes were assessed via individually written reports discussing the disease process, interpretation of diagnostic results and, recommendations and rationales for therapeutic interventions. Results: This approach yielded high quality student reports that conformed to antimicrobial prescription guidelines with consistently high summative assessment scores. Mean scores for the final report in this learning activity were: 82 ± 12%, 80 ± 12% and 80 ± 11% for 2015, 2016 and 2017 cohorts respectively; over the same time period, 98 ± 1% of students indicated that these learning activities facilitated the development of confidence, professional knowledge and skills. Discussion: This was a consistent approach for integrating principles of veterinary pharmacology and microbiology in clinical disciplines. These data illustrate the benefit of a systematic application of a cooperative, case-based learning and teaching model in integrating pre-clinical and clinical disciplines in a bachelor of veterinary science course

Assessment of urban solid waste logistics systems: the case of Kampala, Uganda

Many cities, especially in developing countries, are facing challenges in the management of solid waste. The aim of the study was to develop effective logistics systems for solid waste management in urban areas of developing countries, with a specific focus on Kampala, Uganda. This thesis contains an assessment of the reverse logistics systems that enable effective recapturing of valuable products from urban solid waste. The study mapped the waste collection systems in Kampala using a geographic information system (GIS), i.e. ArcGIS software, and examined the existing models of waste collection to the final disposal destinations. It was found that food and yard wastes constitute 92.7 % of the waste that reaches landfills in Kampala. Recyclables and other special wastes constitute only 7.3 % of the total waste. The generation rate of solid wastes on average from poor areas, upscale wealthier areas, business centres and market areas was 582, 169, 105 and 90 tonnes/day respectively. The study optimised travel distances, number of vehicles and collection time, while maximising total waste collection for environmental sustainability. Results showed that, an increase from a 6-tonne truck to a 10-tonne one reduced the travel distance by 39 %, while an increase from a 10-tonne truck to an 18-tonne truck reduced the travel distance by 34 % considering the current 40 % waste collection. Suggestions regarding the best waste collection routes and a suitable vehicle fleet and capacity to be used by Kampala Capital City Authority (KCCA) have been provided in this study. The research study further developed an overview of reverse logistics at the Kiteezi landfill. The study analysed in detail the collection, re-processing, re-distribution and final markets of these products into a reversed supply chain network of products delivered to the landfill. Of the products at the Kiteezi landfill, 14 % was channelled into the reverse chain, 63 % could be included in the distribution chain but were left out and disposed of while the remaining 23 % was buried straightaway. The main conclusion of the work was that solid waste management in Kampala is characterised by inefficient collection methods, insufficient coverage of the collection system and improper disposal of municipal solid waste. The existing system pertaining to reverse logistics suffers from unfavourable economics and legislative, technical and operational constraints that affect the recycling rate in Kampala compared to developed countries. This study presented large-scale data that can be used to improve solid waste management in other cities in developing countries

Maintaining normal blood pressure in a renovascular stenosis model of hypertension in adult Lewis rats: putative physiological modulation of the renin-angiotensin system

Prolonged unilateral renal artery stenosis using a Goldblatt Two-Kidney-One Clip (2K1C) technique is a validated scientific approach to inducing experimental hypertension in laboratory animals. This patho-physiological modulation is associated with the activation of the renin-angiotensin system with increased renin and angiotensin II release to initiate hypertension. This mode of experimental hypertension has been demonstrated for many rat strains including Sprague Dawley, Brown Norway and Wistar Kyoto but has not been fully characterized in syngeneic Lewis rats. The objective of this study was to develop and characterize a unilateral renal artery stenosis model of hypertension in adult male Lewis rats using a 2K1C method for hypertension studies in our laboratory. Thirty animals were randomly assigned to two groups to undergo sham or the 2K1C surgical procedure under isoflurane-induced anesthesia; approval was granted by our institutional animal ethics committee (A2404, n =15 per group). Hemodynamic parameters including heart rate; systolic and diastolic blood pressure were monitored weekly for 6 weeks, using a volume-pressure tail cuff and a CODA pressure computer. Plasma concentrations of critical biomarkers of the renin-angiotensin system were quantified using standard ELISA assays and renal histopathological changes were assessed microscopically. Unilateral renal arterial stenosis caused marked renal atrophy, glomerulosclerosis and interstitial fibrosis, but a compensatory renal hypertrophy was observed in contralateral kidneys. Interestingly, unilateral renal arterial stenosis failed to induce hypertension over the six week period. The resistance to develop hypertension in the 2K1C group was associated with a significant decrease in total plasma renin (P = 0.023), an increase in the ratio of plasma angiotensin (1-7) to total renin (P = 0.034) and a decrease in the ratio of angiotensin II to total renin (P = 0.013). There were no changes in plasma electrolytes, glucose, creatinine or urea. These data demonstrate that adult male Lewis rats may have physiologically modulated the activation of the renin-angiotensin system to maintain a homeostatic balance and resist hypertension following unilateral renal artery stenosis

Testudines as sentinels for monitoring the dissemination of antibiotic resistance in marine environments: an integrative review

Dissemination of antibiotic resistance (AR) in marine environments is a global concern with a propensity to affect public health and many ecosystems worldwide. We evaluated the use of sea turtles as sentinel species for monitoring AR in marine environments. In this field, antibiotic resistant bacteria have been commonly identified by using standard culture and sensitivity tests, leading to an overrepresentation of specific, culturable bacterial classes in the available literature. AR was detected against all major antibiotic classes, but the highest cumulative global frequency of resistance in all represented geographical sites was against the beta-lactam class by a two-fold difference compared to all other antibiotics. Wastewater facilities and turtle rehabilitation centres were associated with higher incidences of multidrug-resistant bacteria (MDRB) accounting for an average of 58% and 49% of resistant isolates, respectively. Furthermore, a relatively similar prevalence of MDRB was seen in all studied locations. These data suggest that anthropogenically driven selection pressures for the development of AR in sea turtles and marine environments are relatively similar worldwide. There is a need, however, to establish direct demonstrable associations between AR in sea turtles in their respective marine environments with wastewater facilities and other anthropogenic activities worldwide

Are interactions between epicardial adipose tissue, cardiac fibroblasts and cardiac myocytes instrumental in atrial fibrosis and atrial fibrillation?

Atrial fibrillation is very common among the elderly and/or obese. While myocardial fibrosis is associated with atrial fibrillation, the exact mechanisms within atrial myocytes and surrounding non-myocytes are not fully understood. This review considers the potential roles of myocardial fibroblasts and myofibroblasts in fibrosis and modulating myocyte electrophysiology through electrotonic interactions. Coupling with (myo)fibroblasts in vitro and in silico prolonged myocyte action potential duration and caused resting depolarization; an optogenetic study has verified in vivo that fibroblasts depolarized when coupled myocytes produced action potentials. This review also introduces another non-myocyte which may modulate both myocardial (myo)fibroblasts and myocytes: epicardial adipose tissue. Epicardial adipocytes are in intimate contact with myocytes and (myo)fibroblasts and may infiltrate the myocardium. Adipocytes secrete numerous adipokines which modulate (myo)fibroblast and myocyte physiology. These adipokines are protective in healthy hearts, preventing inflammation and fibrosis. However, adipokines secreted from adipocytes may switch to pro-inflammatory and pro-fibrotic, associated with reactive oxygen species generation. Pro-fibrotic adipokines stimulate myofibroblast differentiation, causing pronounced fibrosis in the epicardial adipose tissue and the myocardium. Adipose tissue also influences myocyte electrophysiology, via the adipokines and/or through electrotonic interactions. Deeper understanding of the interactions between myocytes and non-myocytes is important to understand and manage atrial fibrillation

Nutritional enhancement of health beneficial omega-3 long-chain polyunsaturated fatty acids in the muscle, liver, kidney, and heart of Tattykeel Australian White MARGRA lambs fed pellets fortified with omega-3 oil in a feedlot system.

The aim of this research was to evaluate the nutritional enhancement of omega-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA) composition of edible lamb Longissimus thoracis et lumborum muscle, heart, kidney, and liver in response to dietary supplementation of lot-fed lambs with or without omega-3 oil fortified pellets. The hypothesis tested was that fortifying feedlot pellets with omega-3 oil will enhance the human health beneficial n-3 LC-PUFA composition of edible lamb muscle tissue and organs. Seventy-five Tattykeel Australian White lambs exclusive to the MARGRA brand, with an average body weight of 30 kg at six months of age, were randomly assigned to the following three dietary treatments of 25 lambs each, and lot-fed as a cohort for 47 days in a completely randomized experimental design: (1) Control grain pellets without oil plus hay; (2) Omega-3 oil fortified grain pellets plus hay; and (3) Commercial whole grain pellets plus hay. All lambs had ad libitum access to the basal hay diet and water. Post-slaughter fatty acid composition of the Longissimus thoracis et lumborum muscle, liver, kidney, and heart were determined using thee gas chromatography–mass spectrophotometry technique. Results indicated significant variations (p < 0.05) in fatty acid profiles between tissues and organs. Omega-3 oil fortified pellets significantly (p < 0.05) increased ≥C20 n-3 LC-PUFA (C20:5n-3 eicosapentaenoate, EPA + C22:5n3 docosapentaenoate, DPA + C22:6n3 docosahexanoate DHA); C18:3n-3 alpha-linolenate, ALA; C18:2 conjugated linoleic acid, CLA; total monounsaturated fatty acids, MUFA; polyunsaturated fatty acids, PUFA contents; and reduced the ratio of omega-6 to omega-3 fatty acids in all lamb organs and tissues without impacting shelf-life. The findings demonstrate that the inclusion of omega-3 oil in feedlot diets of lambs enhances the human health beneficial omega-3 long-chain polyunsaturated fatty acid profiles of edible muscle tissue and organs without compromising meat quality

Angiopoietin-2 attenuates angiotensin II-induced aortic aneurysm and atherosclerosis in apolipoprotein E-deficient mice

Angiogenesis and inflammation are implicated in aortic aneurysm and atherosclerosis and regulated by angiopoietin-2 (Angpt2). The effect of Angpt2 administration on experimental aortic aneurysm and atherosclerosis was examined. Six-month-old male apolipoprotein E deficient (ApoE⁻/⁻) mice were infused with angiotensin II (AngII) and administered subcutaneous human Fc-protein (control) or recombinant Angpt2 (rAngpt2) over 14 days. Administration of rAngpt2 significantly inhibited AngII-induced aortic dilatation and rupture of the suprarenal aorta (SRA), and development of atherosclerosis within the aortic arch. These effects were blood pressure and plasma lipoprotein independent and associated with Tie2 activation and down-regulation of monocyte chemotactic protein-1 (MCP-1) within the SRA. Plasma concentrations of MCP-1 and interleukin-6 were significantly lower in mice receiving rAngpt2. Immunostaining for the monocyte/macrophage marker MOMA-2 and the angiogenesis marker CD31 within the SRA were less in mice receiving rAngpt2 than controls. The percentage of inflammatory (Ly6Cʰⁱ) monocytes within the bone marrow was increased while that in peripheral blood was decreased by rAngpt2 administration. In conclusion, administration of rAngpt2 attenuated angiotensin II-induced aortic aneurysm and atherosclerosis in ApoE⁻/⁻ mice associated with reduced aortic inflammation and angiogenesis. Up-regulation of Angpt2 may have potential therapeutic value in patients with aortic aneurysm and atherosclerosis

MARGRA lamb eating quality and human health-promoting omega-3 long-chain polyunsaturated fatty acid profiles of Tattykeel Australian White sheep: linebreeding and gender effects

Health-conscious consumers increasingly demand healthier, tastier, and more nutritious meat, hence the continuous need to meet market specifications and demand for high-quality lamb. We evaluated the longissimus dorsi muscle of 147 Tattykeel Australian White (TAW) sheep fed on antioxidant-rich ryegrass pastures exclusive to MAGRA lamb brand for meat eating quality parameters of intramuscular fat (IMF) content, fat melting point (FMP) and omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA). The aim was to assess the impact of linebreeding and gender on pasture-fed lamb eating quality and to test the hypothesis that variation in healthy lamb eating quality is a function of lamb gender and not its antioxidant status or inbreeding coefficient (IC). After solid-phase extraction and purification, phenolics and antioxidant enzyme activities were analysed by high-performance liquid chromatography and mass spectrometry. IMF and fatty acid composition were determined using solvent extraction and gas chromatography, respectively. IC was classified into low (0–5%), medium (6–10%) and high (>10%) and ranged from 0–15.6%. FMP and IMF ranged from 28 to 39 °C and 3.4% to 8.2%, with overall means of 34.6 ± 2.3 °C and 4.4 ± 0.2%, respectively, and n-3 LC-PUFA ranged from “source” to “good source” levels of 33–69 mg/100 g. Ewes had significantly (P ˂ 0.0001) higher IMF, C22:5n-3 (DPA), C22:6n-3 (DHA), C18:3n-6, C20:3, C22:4n-6, C22:5n-6, total monounsaturated (MUFA), PUFA and Σn-3 fatty acids and lower total saturated fatty acids (SFA) and FMP, than rams. As IC increased, there were no differences in FMP and IMF. Folin–Ciocalteu total phenolics, ferric reducing antioxidant power and antioxidant activities of glutathione peroxidase, catalase and superoxide dismutase enzymes did not differ by either gender or IC. This study provides evidence that IC is inconsequential in affecting antioxidant status, IMF, FMP and n-3 LC-PUFA in linebred and pasture-fed TAW sheep because the observed variation in individual fatty acids was mainly driven by gender differences between ewes and rams, hence the need to accept the tested hypothesis. This finding reinforces the consistent healthy eating quality of MARGRA lamb brand from TAW sheep regardless of its linebred origin