Susan Hall

Cellular Effects of Pyocyanin, a Secreted Virulence Factor of Pseudomonas aeruginosa

Pyocyanin has recently emerged as an important virulence factor produced by Pseudomonas aeruginosa. The redox-active tricyclic zwitterion has been shown to have a number of potential effects on various organ systems in vitro, including the respiratory, cardiovascular, urological, and central nervous systems. It has been shown that a large number of the effects to these systems are via the formation of reactive oxygen species. The limitations of studies are, to date, focused on the localized effect of the release of pyocyanin (PCN). It has been postulated that, given its chemical properties, PCN is able to readily cross biological membranes, however studies have yet to be undertaken to evaluate this effect. This review highlights the possible manifestations of PCN exposure; however, most studies to date are in vitro. Further high quality in vivo studies are needed to fully assess the physiological manifestations of PCN exposure on the various body systems.

A review of the bioactivity of coffee, caffeine and key coffee constituents on inflammatory responses linked to depression

Coffee is a widely consumed beverage containing numerous biologically active constituents predominantly belonging to the polyphenol and alkaloid classes. It has been established that coffee has a beneficial effect on numerous disease states including depression. A number of prospective and retrospective cohort studies have assessed the effects of coffee consumption on the relative risk of developing major depressive disorder in humans. These studies have identified an inverse relationship between the consumption of caffeinated coffee and the risk of developing depression. Caffeine, chlorogenic acid, ferulic acid and caffeic acid, all important constituents of coffee, have been shown to possess biological activities that highlight a possible mechanistic link to the pathology of depression. This review aims to assess the evidence from the biological evaluation of these constituents of coffee on markers of inflammation associated with depression in in vitro and in vivo models of inflammation, neuroinflammation and depression. The ability of bioactive coffee constituents to modulate the parameters of neuroinflammation has been shown with caffeine having strong antioxidant properties in vitro, chlorogenic acid and caffeic acid having strong anti-inflammatory and antioxidant properties in vitro and ferulic acid having activities in in vivo animal models of depression.

Caffeine consumption around an exercise bout: effects on energy expenditure, energy intake, and exercise enjoyment

Combining an exercise and nutritional intervention is arguably the optimal method of creating energy imbalance for weight loss. This study sought to determine whether combining exercise and caffeine supplementation was more effective for promoting acute energy deficits and manipulations to substrate metabolism than exercise alone. Fourteen recreationally active participants (mean ± SD body mass index: 22.7 ± 2.6 kg/m2) completed a resting control trial (CON), a placebo exercise trial (EX), and a caffeine exercise trial (EX+CAF, 2 × 3 mg/kg of caffeine 90 min before and 30 min after exercise) in a randomized, double-blinded design. Trials were 4 h in duration with 1 h of rest, 1 h of cycling at ∼65% power at maximum O2 consumption or rest, and a 2-h recovery. Gas exchange, appetite perceptions, and blood samples were obtained periodically. Two hours after exercise, participants were offered an ad libitum test meal where energy and macronutrient intake were recorded. EX+CAF resulted in significantly greater energy expenditure and fat oxidation compared with EX (+250 kJ; +10.4 g) and CON (+3,126 kJ; +29.7 g) (P < 0.05). A trend for reduced energy and fat intake compared with CON (-718 kJ; -8 g) (P = 0.055) was observed. Consequently, EX+CAF created a greater energy deficit (P < 0.05). Caffeine also led to exercise being perceived as less difficult and more enjoyable (P < 0.05). Combining caffeine with exercise creates a greater acute energy deficit, and the implications of this protocol for weight loss or maintenance over longer periods of time in overweight/obese populations should be further investigated.

Protection against radiotherapy-induced toxicity

Radiation therapy is a highly utilized therapy in the treatment of malignancies with up to 60% of cancer patients receiving radiation therapy as a part of their treatment regimen. Radiation therapy does, however, cause a wide range of adverse effects that can be severe and cause permanent damage to the patient. In an attempt to minimize these effects, a small number of compounds have been identified and are in use clinically for the prevention and treatment of radiation associated toxicities. Furthermore, there are a number of emerging therapies being developed for use as agents that protect against radiation-induced toxicities. The aim of this review was to evaluate and summarise the evidence that exists for both the known radioprotectant agents and the agents that show promise as future radioprotectant agents.

Caffeic acid attenuates lipopolysaccharide-induced sickness behaviour and neuroinflammation in mice

Accumulating data links inflammation, oxidative stress and immune system in the pathophysiology of major depressive disorders. Sickness behaviour is a set of behavioural changes that develop during infection, eventually leading to decrease in mobility and depressed behaviour. Lipopolysaccharide (LPS) induces a depression-like state in animals that mimics sickness behaviour. Caffeic acid, a naturally occurring polyphenol, possesses antioxidant and anti-inflammatory properties. The present study was designed to explore the potential of caffeic acid against LPS-induced sickness behaviour in mice. Caffeic acid (30mg/kg) and imipramine (15mg/kg) were administered orally one hour prior to LPS (1.5mg/kg) challenge. Behavioural assessment was carried out between 1 and 2h and blood samples were collected at 3h post-LPS injection. Additionally, cytokines (brain and serum) and brain oxidative stress markers were estimated. LPS increased the systemic and brain cytokine levels, altered the anti-oxidant defence and produced key signs of sickness behaviour in animals. Caffeic acid treatment significantly reduced the LPS-induced changes, including reduced expression of inflammatory markers in serum and whole brain. Caffeic acid also exerted an anti-oxidant effect, which was evident from the decreased levels of oxidative stress markers in whole brain. Our data suggests that caffeic acid can prevent the neuroinflammation-induced acute and probably the long term neurodegenerative changes.

Indoleamine-2,3-dioxygenase elevated in tumor-initiating cells is suppressed by mitocans.

Tumor-initiating cells (TICs) often survive therapy and give rise to second-line tumors. We tested the plausibility of sphere cultures as models of TICs. Microarray data and microRNA data analysis confirmed the validity of spheres as models of TICs for breast and prostate cancer as well as mesothelioma cell lines. Microarray data analysis revealed the Trp pathway as the only pathway upregulated significantly in all types of studied TICs, with increased levels of indoleamine-2,3-dioxygenase-1 (IDO1), the rate-limiting enzyme of Trp metabolism along the kynurenine pathway. All types of TICs also expressed higher levels of the Trp uptake system consisting of CD98 and LAT1 with functional consequences. IDO1 expression was regulated via both transcriptional and posttranscriptional mechanisms, depending on the cancer type. Serial transplantation of TICs in mice resulted in gradually increased IDO1. Mitocans, represented by α-tocopheryl succinate and mitochondrially targeted vitamin E succinate (MitoVES), suppressed IDO1 in TICs. MitoVES suppressed IDO1 in TICs with functional mitochondrial complex II, involving transcriptional and posttranscriptional mechanisms. IDO1 increase and its suppression by VE analogues were replicated in TICs from primary human glioblastomas. Our work indicates that IDO1 is increased in TICs and that mitocans suppress the protein.

Modulation of chemotherapy-induced cytotoxicity in SH-SY5Y neuroblastoma cells by caffeine and chlorogenic acid

Abstract Chemotherapy is an important treatment modality for malignancy but is limited by significant toxicity and it susceptibility to numerous drug interactions. While the interacting effects with medications are well known, there is limited evidence on the interaction with commonly consumed food and natural products. The aim of this study was to evaluate the bioactive constituents of coffee (caffeine and chlorogenic acid) on the cytotoxicity of doxorubicin, gemcitabine, and paclitaxel in vitro. Pretreatment with caffeine (100 nM and 10 μM) sensitized SH-SY5Y cells to doxorubicin-induced toxicity and increased apoptosis and sensitized PC3 cells to gemcitabine-induced toxicity. Pretreatment with 10 μM caffeine decreased total cell reactive oxygen species (ROS) production but increased mitochondrial ROS production. In contrast, caffeine (10 nM and 10 μM) protected cells against gemcitabine-induced toxicity and apoptosis. Similarly, 1 μM and 10 μM caffeine protected cells against paclitaxel-induced toxicity and mitochondrial ROS production. Chlorogenic acid had no effect on chemotherapy-induced toxicity in SH-SY5Y cells. In conclusion, this study provides preliminary evidence that caffeine, not chlorogenic acid, modulates the cytotoxicity of doxorubicin, gemcitabine, and paclitaxel in SH-SY5Y cells via different mechanisms.

Effect of meal glycemic load and caffeine consumption on prolonged monotonous driving performance

OBJECTIVE Monotonous driving involves low levels of stimulation and high levels of repetition and is essentially an exercise in sustained attention and vigilance. The aim of this study was to determine the effects of consuming a high or low glycemic load meal on prolonged monotonous driving performance. The effect of consuming caffeine with a high glycemic load meal was also examined. METHOD Ten healthy, non-diabetic participants (7 males, age 51±7yrs, mean±SD) completed a repeated measures investigation involving 3 experimental trials. On separate occasions, participants were provided one of three treatments prior to undertaking a 90min computer-based simulated drive. The 3 treatment conditions involved consuming: (1) a low glycemic load meal+placebo capsules (LGL), (2) a high glycemic load meal+placebo capsules (HGL) and (3) a high glycemic load meal+caffeine capsules (3mgkg-1 body weight) (CAF). Measures of driving performance included lateral (standard deviation of lane position (SDLP), average lane position (AVLP), total number of lane crossings (LC)) and longitudinal (average speed (AVSP) and standard deviation of speed (SDSP)) vehicle control parameters. Blood glucose levels, plasma caffeine concentrations and subjective ratings of sleepiness, alertness, mood, hunger and simulator sickness were also collected throughout each trial. RESULT No difference in either lateral or longitudinal vehicle control parameters or subjective ratings were observed between HGL and LGL treatments. A significant reduction in SDLP (0.36±0.20m vs 0.41±0.19m, p=0.004) and LC (34.4±31.4 vs 56.7±31.5, p=0.018) was observed in the CAF trial compared to the HGL trial. However, no differences in AVLP, AVSP and SDSP or subjective ratings were detected between these two trials (p>0.05). CONCLUSION Altering the glycemic load of a breakfast meal had no effect on measures of monotonous driving performance in non-diabetic adults. Individuals planning to undertake a prolonged monotonous drive following consumption of a morning meal may consider consuming caffeine as a means of improving vehicle control.

A pilot study assessing the value of 3D printed molecular modelling tools for pharmacy student education

BACKGROUND AND PURPOSE Medicinal chemistry and pharmacology are difficult topics to both teach and learn given the complex nature of drug mechanisms and drug-receptor interactions. This highlights the need for innovative teaching methods to deliver this information to students. One such method is through three-dimensional (3D) printing of enzymes and ligands in the teaching of molecular modelling concepts relating to drug-receptor and enzyme interactions be ligands. This type of printing has been shown to be beneficial in several educational settings; however, to our knowledge, its effectiveness in pharmacy, medicinal chemistry and pharmacology learning and teaching is largely unknown. Therefore, the aim of this study was to evaluate pharmacy student perceptions and the educational benefits of 3D printed molecules in molecular modelling with regards to engagement and learning outcomes when used in a drug-target interaction topic. EDUCATIONAL ACTIVITY AND SETTING This aim was achieved through administering students a short questionnaire designed to evaluate their engagement and learning outcomes with students also free to provide comments. FINDINGS This study found that nearly all (>90%) students found the activity was useful in improving both student engagement and learning outcomes. DISCUSSION AND SUMMARY In conclusion, 3D printing may provide an alternative learning activity to help pharmacy students understand the drug-target interaction.

Pyocyanin induces systemic oxidative stress, inflammation and behavioral changes in vivo

Abstract Pyocyanin (PCN) is a virulence factor secreted by Pseudomonas aeruginosa (P. aeruginosa) that has been shown to have numerous toxic effects in both in vitro and in vivo studies. Such toxicities include pro-inflammatory and pro-oxidant mediated responses. It is hypothesized that PCN can cross biological membranes and reach the systemic circulation, but no previous studies have investigated this. The aim of this study was, therefore, to quantify PCN in plasma and assess if systemic responses were occurring after localized intranasal administration in C57BL/6 J mice. This was achieved through the plasma quantification of PCN and assessment of changes to behavior using two commonly used tests, the forced swimming test and the open field test. Furthermore, evidence of systemic oxidative stress and inflammation was measured using malondialdehyde (MDA) and TNF-α post PCN exposure. PCN was found to cross into systemic circulation but in a variable manner. Furthermore, significant increases in plasma TNF-α and MDA (both p < 0.001) were observed along with changes in behavior indicative of systemic inflammatory responses.