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NAD+—dependent histone deacetylases (sirtuins 1–7) have been shown to be involved in various pathophysiological conditions including their involvement in cardiovascular, cancerous, neurodegenerative, immune dysregulation and inflammatory conditions. This study investigates the inflammomodulatory potential of resveratrol (RES), a sirtuin activator and sirtinol (SIR), a sirtuin inhibitor in lipopolysaccharide (LPS)-induced model of sickness behaviour in mice. Male Swiss albino mice were divided into five groups (n = 6) consisting of saline (SAL), LPS, RES, SIR, and fluoxetine (FLU) respectively, each group except LPS was prepared by intraperitoneally (i.p.) administration of SAL (10 mL/kg), RES (50 mg/kg), SIR (2 mg/kg) and FLU (10 mg/kg). Thirty minutes after the treatments, all the groups, except SAL were administered LPS (2 mg/kg, i.p.). The behavioural assays including, open field test, forced swim test, and tail suspension tests were conducted 1 h after LPS challenge. LPS administration significantly reduced the locomotor activity along with inducing a state of high immobility and that was prevented by pretreatment with RES and SIR. Further, various proinflammatory cytokines (TNF-α, IL-6, and IL-1β), and oxidative stress markers (MDA and GSH) were found to be significantly elevated in the brain homogenates after LPS treatment. SIR pretreatment abrogated the LPS-induced neuroinflammatory and oxidative stress changes, whereas RES was only effective in reducing the oxidative stress and TNF-α levels. The results of this study speculate that the role of SIRT modulators in neuroinflammatory conditions could vary with their dose, regimen and chemical properties. Further studies with detailed molecular and pharmacokinetic profiling will be needed to explore their therapeutic potentials.

To achieve malaria eradication, new preventative agents that act differently to front-line treatment drugs are needed. To identify potential chemoprevention starting points we screened a sub-set of the CSIRO Australia Compound Collection for compounds with slow-actionin vitroactivity againstPlasmodium falciparum. This work identifiedN,N-dialkyl-5-alkylsulfonyl-1,3,4-oxadiazol-2-amines as a new antiplas-modial chemotype (e.g.,196 h IC50550 nM) with a different action to delayed-death slow-action drugs. Structure activity relationship analysis of analogues identified multiple compounds with potent and selectivein vitroactivity against drug-sensitive and multi-drug resistantPlasmodiumparasites (e.g.,31and3296 h IC502,500). However subsequent studies in mice with lead compound1, which had the best microsomal stability of the compounds assessed, demonstrated rapid clearance (T1/2<1.6 h) and poor oralin vivoefficacy. This indicates that improvements in the pharmacokinetic profile ofN,N-dialkyl-5-alkylsulfonyl-1,3,4-oxadiazol-2-amines would be needed for the development of this chemotype for malaria chemoprophylaxis.

Neuroinflammation is associated with the development of depression. Deacetylases SIRT1 and SIRT2 are reported to exert neuroprotective effects in aging, neurogenesis, neurodegeneration and neuroinflammation. Therefore, this study aimed to investigate the effects of SIRT1 and SIRT2 modulators on LPS-induced neuroinflammation and neurodegeneration in vitro. To achieve this, HAPI rat microglial cells were pre-treated with the SIRT1 activator resveratrol (0.1–20 µM), the selective SIRT1 inhibitor EX527 (0.1; 1 µM), the dual SIRT1/SIRT2 inhibitor sirtinol (0.1–20 µM) and the SIRT2 inhibitor AGK2 (0.1; 1 µM), prior to exposure with LPS (5 ng/mL) for 20 h. The reference antidepressant drug fluoxetine and the nonsteroidal anti-inflammatory drug ibuprofen were also evaluated in the same paradigm, both at 1 μM. Resveratrol and sirtinol inhibited TNF-α production to a greater degree than either fluoxetine or ibuprofen. Resveratrol, sirtinol, EX527 and AGK2 significantly reduced PGE2 production by up to 100% in microglia. Then, the supernatant was transferred to treat SH-SY5Y cells for 24 h. In all cases, SIRT modulator pretreatment significantly protected undifferentiated SH-SY5Y human neuroblastoma cells from the insult of LPS-stimulated HAPI supernatant by up to 40%. Moreover, resveratrol and sirtinol also showed significantly better neuroprotection than fluoxetine or ibuprofen by up to 83 and 69%, respectively. In differentiated SH-SY5Y cells, only sirtinol (20, 10 µM) and AGK2 (0.1 µM) pretreatment protected the cells from LPS-stimulated HAPI supernatant. This study suggests that SIRT1 and SIRT2 modulators are effective in inhibiting LPS-stimulated production of TNF-α and PGE2 in HAPI microglial cells and protecting SH-SY5Y cells from inflammation. Thus, we provide proof of concept for further investigation of the therapeutic effect of SIRT1 and SIRT2 modulators and combination with current antidepressant medication as a treatment option.

A series of novel furan-2-yl-1H-pyrazoles and their chemical precursors were synthesized and evaluated for their effectiveness at disrupting α-synuclein (α-syn) aggregation in vitro. The compounds were found to inhibit α-syn aggregation with efficacy comparable to the promising drug candidate anle138b. The results of this study indicate that compounds 8b, 8l, and 9f may qualify as secondary leads for the structure-activity relationship studies aimed to identify the suitable compounds for improving the modulatory activity targeted at α-syn self-assembly related to Parkinson's disease.

Depression is a psychiatric disorder that has a significant health burden on patients and their families. Unfortunately, the current antidepressant medications that mainly target monoamine neurotransmitters have limited efficacy. Recent evidence has indicated that neuroinflammation participates in the genesis and development of depression, and interacts with other factors involved in depression. Therefore, exploring effective anti-inflammatory medications could be beneficial for the development of new treatment options for depression. Sirtuins are a unique class of nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases, which have seven members that can affect multiple downstream targets by deacetylation activity. Among these seven members, SIRT1 and SIRT2 have been shown to participate in the pathophysiology of inflammation in numerous studies. Thus, in this short article, we review the association of SIRT1 and SIRT2 activity and depression, and evidence of the effects of SIRT1 and SIRT2 modulators on inflammation in vitro and depressive-like behaviours in vivo.

Objectives Warfarin remains widely used with a time in therapeutic range (TiTR) above 65% recommended for best outcomes. Patients not achieving or maintaining this warfarin control may be better suited to alternate anticoagulants. Despite this, there is limited data defining a suitable trial time in patients initiating warfarin therapy, therefore the aim of this study was to determine the mean time to stable therapeutic range (TtSTR). Materials and Methods Retrospective data was collected for patients with atrial fibrillation enrolled in a dedicated warfarin program at a private pathology practice within 7 days of warfarin initiation. TiTR at specified timepoints was calculated and median TtSTR determined as defined by TiTR ≥ 65% over three months. Comparisons were made of populations with TtSTR above or below the median. Results The 566 patients included in the study had a mean TiTR of 64.9±16.5% at month three and median TtSTR of six months. Patients with TtSTR≤6 months achieved a mean TiTR of 68.9±12.8% at month two and maintained a TiTR over 75% from month 3 to 24. Patients with a TtSTR>6 months obtained a TiTR of 66.4±10.6% at month nine and continued to achieve lower TiTR throughout the 24 months study period. Conclusions A majority of patients can achieve a stable TiTR above 65% within six months so review at six to nine months is likely to be a good indicator of warfarin control and to determine if patients should continue warfarin or switch to alternate anticoagulant therapy.

A series of novel furan-2-yl-1

3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins) have been associated with conflicting effects within the central nervous system (CNS), with underlying mechanisms remaining unclear. Although differences between individual statins' CNS effects have been reported clinically, few studies to date have compared multiple statins' neuroprotective effects. This study aimed to compare six statins (atorvastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin; 0–100 μM) using an in vitro model of lipopolysaccharide (LPS)-induced neuroinflammation and subsequent neurodegeneration. To achieve this, HAPI microglia were treated with LPS (0.1 μg/mL; 24 h), resulting in increased reactive oxygen species (ROS), nitric oxide, and IL-1β, TNF-α and PGE2 release. Conditioned media (“HAPI-CM”) was then transferred to SH-SY5Y neuroblastoma cells, and effects on cellular viability, mitochondrial membrane permeability, apoptosis, autophagy and ROS production assessed. Of the statins investigated, only atorvastatin, pravastatin and rosuvastatin protected SH-SY5Y cells from LPS-induced decreases in cellular viability; this appeared mediated through reduced caspase 3/7 activation and was associated with decreased IL-1β (atorvastatin, pravastatin) and/or TNF-α (atorvastatin, pravastatin, rosuvastatin). Only pravastatin conferred protection at all tested concentrations. ROS production and autophagic vacuole formation was decreased by all statins, suggesting these two mechanisms are unlikely to be sole mediators of neuroprotection seen with selected statins. Ultimately, our model suggests that despite all statins reducing microglial inflammation, subsequent effects on neuronal viability and cell death signalling pathways varies between statins. Our findings highlight the need to consider individual statins as inducing discrete pharmacological effects within the CNS in future in vitro/in vivo studies and in clinical practice.

Background: Non-adherence to prescribed medicines is linked to adverse health outcomes in people living with chronic health conditions (CHCs). Multiple factors are known to contribute to non-adherence to medicines including polypharmacy, demographic features and disease and health systems. Both non-prescription and prescription medicines contribute to polypharmacy; however, there is limited data on the influence of non-prescription medicines to non-adherence. Aim: Therefore, the aim of the study was to investigate the influence of non-prescription medicines to non-adherence in an Australian population. Methods: Data from the 2016 National Survey of a random sample of Australian adult residents were utilised in this study to investigate factors associated with non-adherence. Descriptive statistics, chi(2), regression and generalised linear models were used to assess the relationships between variables of interest. Narrative response and comments were used to provide further insight. Results: This study recruited 1217 participants to explore factors associated with non-adherence to medicines. Weak but statistically significant correlations were identified showing the number of CHCs, patient's age, number of prescription medicines, number of non-prescription medicines and total number of medicines associated with non-adherence. Discussion: The findings suggest that people living with CHCs and taking multiple medicines, including non-prescription medicines, are likely to be non-adherent to prescription medicines. This study shows the possible involvement of non-prescription medicines in contributing to non-adherence in an Australian population and suggests that future studies with a broader demographic are warranted.

Neurodegenerative disease refers to a range of chronic and progressive disorders that are characterized by dysfunction and loss of neurons. Neurodegeneration involves protein misfolding, oxidative injury, impaired mitochondrial function, neurotrophin deficiency and may also involve neuroinflammation. The sirtuin family of proteins plays a key role in this process suggesting that modulation of sirtuin can modify disease progression. This review examines experimental and clinical evidence relating to the potential role of SIRT1 and SIRT2, and their modulators in neurodegenerative diseases. Both neuroprotective effects and negative effects of SIRT1 activators, SIRT1 inhibitors and SIRT2 activators are discussed in a range of different disease models, including in vitro and in vivo Alzheimer's disease (AD), Parkinson's disease (PD), Huntingdon's disease (HD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS). This highlights the potential of SIRT1 and SIRT 2 modulators as potential therapeutic agents. However, there is a paucity of clinical trials related to the effects of selective SIRT1 modulators, selective SIRT2 modulators or dual SIRT1/2 modulators on neuroinflammation and subsequent neurodegeneration.

Numerous post-translational modifications (PTMs) of the Parkinson's disease (PD) associated α-synuclein (α-syn) protein have been recognised to play critical roles in disease aetiology. Indeed, dysregulated phosphorylation and proteolysis are thought to modulate α-syn aggregation and disease progression. Among the PTMs, enzymatic glycosylation with N-acetylglucosamine (GlcNAc) onto the protein's hydroxylated amino acid residues is reported to deliver protective effects against its pathogenic processing. This modification has been reported to alter its pathogenic self-assembly. As such, manipulation of the protein's O-GlcNAcylation status has been proposed to offer a PD therapeutic route. However, targeting upstream cellular processes can lead to mechanism-based toxicity as the enzymes governing O-GlcNAc cycling modify thousands of acceptor substrates. Small glycopeptides that couple the protective effects of O-GlcNAc with the selectivity of recognition sequences may prove useful tools to modulate protein aggregation. Here we discuss efforts to probe the effects of various O-GlcNAc modified peptides on wild-type α-synuclein aggregation.

Post-translational modifications (PTMs) of proteins are becoming the focus of intense research due to their implications in a broad spectrum of neurodegenerative diseases. Various PTMs have been identified to alter the toxic profiles of proteins which play critical roles in disease etiology. In Alzheimer's disease (AD), dysregulated phosphorylation is reported to promote pathogenic processing of the microtubule-associated tau protein. Among the PTMs, the enzymatic addition of N-acetyl-d-glucosamine (GlcNAc) residues to Ser/Thr residues is reported to deliver protective effects against the pathogenic processing of both amyloid precursor protein (APP) and tau. Modification of tau with as few as one single O-GlcNAc residue inhibits its toxic self-assembly. This modification also has the same effect on the assembly of the Parkinson's disease (PD) associated α-synuclein (ASyn) protein. In fact, O-GlcNAcylation ( O-linked GlcNAc modification) affects the processing of numerous proteins implicated in AD, PD, amyotrophic lateral sclerosis (ALS), and Huntington's disease (HD) in a similar manner. As such, manipulation of a protein's O-GlcNAcylation status has been proposed to offer therapeutic routes toward addressing multiple neurodegenerative pathologies. Here we review the various effects that O-GlcNAc modification, and its modulated expression, have on pathogenically significant proteins involved in neurodegenerative disease.

Background Warfarin reduces stroke risk in atrial fibrillation (AF) patients but requires ongoing monitoring. Time in therapeutic range (TTR) is used as a measure of warfarin control, with a TTR less than 60% associated with adverse patient outcomes. The Sex, Age, Medical history, Treatment, Tobacco use, Race (SAMe-TT 2 R 2 ) score has been identified as a model able to predict warfarin control, but this has been tested in mainly Caucasian populations. Therefore, the aim of this study was to determine the ability of the SAMe-TT 2 R 2 score to predict warfarin control in a Singaporean population consisting of Chinese, Malay, and Indian race. Methods Retrospective data were collected from the National Heart Centre Singapore for AF patients receiving warfarin between January and June 2014. The TTR and the SAMe-TT 2 R 2 score were calculated for each patient. Results The 1137 non-valvular AF patients had a mean TTR of 58.0 ± 34.3% and a median SAMe-TT 2 R 2 score of 3. The categorized SAMe-TT 2 R 2 scores (2 versus >2) showed a significant reduction in mean TTR for the entire population (63.2% versus 55.8%, P = .0004) and also when categorized according to race for Chinese (62.7% versus 56.9%, P = .0075) and Malay (68.4% versus 50.6%, P = .0131) populations. Conclusion The SAMe-TT 2 R 2 tool is effective in predicting warfarin control in a Singaporean population as patients with a score greater than 2 had poor control. The minimum score for non-Caucasian patients is 2; thus, in these patients, the presence of any additional risk factors identified in the SAMe-TT 2 R 2 tool categorizes them as unlikely to achieve adequate warfarin control and possible candidates for alternative anticoagulants.

Objective To investigate older patients' encounters with general practice registrars (GPRs) to inform training and clinical practice. Method Cross-sectional analysis of data from GPR consultations across five regional training providers in Australia. Data were analysed using simple and multiple logistic regression models. Results Our analysis included details of 118 831 consultations, 20 555 (17.6%, 95% CI 17.4–17.8) with patients aged ≥65 years. Older patient encounters had an increased likelihood of including chronic disease (OR 1.77, 95% CI 1.70, 1.86) and more problems (OR 1.24, 95% CI 1.20, 1.27). However, in-consultation information or advice was less likely to be sought (OR 0.92, 95% CI 0.88, 0.97), and consultations were briefer (OR 0.99, 95% CI 0.99, 1.00). Conclusion Our results suggest relatively limited GPR exposure to older patients coupled with less complex consultations than expected. Solutions will need to be carefully constructed not only to increase caseloads, but also to address training and supervision concerns.

Background: Breath alcohol responses may be affected by the presence of carbohydrate (CHO) in a beverage. This study investigated the impact of consuming alcohol with mixers containing various doses of CHO or an artificial sweetener on breath alcohol concentration (BrAC), ratings of intoxication and impairment, and cognitive performance in females. Methods: Twenty-six females (age 25.1 ± 0.7 years, mean ± standard deviation) completed a crossover study involving 4 trials. A dose of alcohol was consumed in each trial mixed with water (W), artificial sweetener (150 ± 1 mg aspartame [AS]), or CHO (15 g sucrose [15CHO] and 50 g sucrose [50CHO]). BrAC was sampled for 210 minutes following beverage ingestion and analyzed for peak BrAC and other parameters using WinNonlin noncompartmental pharmacokinetic modeling (cmax, tmax, area under the curve to the last measured time point [AUClast]). An objective measure of cognitive performance was assessed using a 4-choice reaction time (CRT) task. Estimation of BrAC, self-reported ratings of intoxication, and willingness to drive were recorded. Results: Mean peak BrAC was reduced in a dose–response manner when alcohol was consumed with CHO compared to both W and AS treatments (W: 0.054 ± 0.015%, AS: 0.052 ± 0.011%, 15CHO: 0.049 ± 0.008%, 50CHO: 0.038 ± 0.007%). No difference in peak BrAC was observed between W and AS treatments. WinNonlin parameters revealed significant differences in cmax and AUClast (W: 4.80 ± 1.12 g/dl/h, AS: 4.61 ± 0.92 g/dl/h, 15CHO: 4.10 ± 0.86 g/dl/h, 50CHO: 3.11 ± 0.58 g/dL/h) when CHO-containing beverages were consumed compared to W and AS treatments. No difference in tmax or CRT was observed between treatments. Participants were able to detect subtle differences in peak BrAC and reported greater ability to drive after consuming 50CHO compared to W. However, participant's willingness to drive and CRT did not differ between treatments. Conclusions: Consuming alcohol with CHO-containing mixers attenuates peak BrAC and reduces total alcohol exposure in a dose–response manner compared to drinks containing artificial sweetener or no additives. The effect of adding CHO to alcoholic beverages may translate to reduced risk of alcohol-related harms.

Background Warfarin is widely prescribed to decrease the risk of stroke in atrial fibrillation (AF) patients. Due to patient variability in response, regular monitoring is required, and time in therapeutic range (TTR) used to indicate quality of warfarin control with a TTR>60% is recommended. Recently, an Australian Government review of anticoagulants identified the need to establish current warfarin control and determine the potential place of the newer oral anticoagulants. Aim To determine warfarin control by a pathology practice in Queensland, Australia and identify factors influencing TTR. Methods Retrospective data were collected from Sullivan Nicolaides Pathology, a major pathology practice offering a warfarin care programme in Australia. Patients enrolled in their programme as of September 2014 were included in the study. TTR was calculated using INR test results, and test dates using the Rosendaal method with mean patient TTR were used for analysis and comparison. Exclusions were target therapeutic range outside 2.0–3.0, less than two INR tests and programme treatment time of less than 30 days. Results The eligible 3692 AF patients had 73.6% of INR tests within the therapeutic range. The mean TTR was 81%, with 97% of patients above a TTR of 60%. TTR was not significantly influenced by age, gender or socioeconomic factors. Conclusions The observed mean TTR of over 80% is superior to the minimum recommended threshold of 60%. The TTR achieved by the Queensland pathology practice demonstrates that dedicated warfarin programmes can produce high-quality warfarin care, ensuring the full benefit of warfarin for Australian patients.

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Alzheimer's disease (AD) is a progressive neurodegenerative disorder accounting for 60-80% of dementia cases. For many years, AD causality was attributed to amyloid-β (Aβ) aggregated species. Recently, multiple therapies that target Aβ aggregation have failed in clinical trials, since Aβ aggregation is found in AD and healthy patients. Attention has therefore shifted toward the aggregation of the tau protein as a major driver of AD. Numerous inhibitors of tau-based pathology have recently been developed. Diagnosis of AD has shifted from measuring late stage senile plaques to early stage biomarkers, amyloid-β and tau monomers and oligomeric assemblies. Synthetic peptides and some derivative structures are being explored for use as theranostic tools as they possess the capacity both to bind the biomarkers and to inhibit their pathological self-assembly. Several studies have demonstrated that O-linked glycoside addition can significantly alter amyloid aggregation kinetics. Furthermore, natural O-glycosylation of amyloid-forming proteins, including amyloid precursor protein (APP), tau, and α-synuclein, promotes alternative nonamyloidogenic processing pathways. As such, glycopeptides and related peptidomimetics are being investigated within the AD field. Here we review advancements made in the last 5 years, as well as the arrival of sugar-based derivatives.

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.

Background Australian community pharmacists are well placed to provide medication-related support and contribute to optimization of outcomes for mental health consumers and their caregivers. However, little is known about the actual competencies of community pharmacists to provide this care. To determine how graduates are being prepared to competently assist mental health consumers and their caregivers, an exploration of the mental health content of university pharmacy programs that set the foundation for pharmacists’ professional roles is needed. Aim To investigate the mental health content of accredited Australian pharmacy qualifying programs. Method A review of publically available online profile information for accredited degree programs was conducted, and program coordinators from the 18 accredited pharmacy degree programs providers in Australia were surveyed. Results Mental health education is embedded in core subjects such as pharmacology, pharmacotherapy, and pharmacy practice. Multiple options are employed to deliver mental health teaching, including lectures, workshops, and experiential learning. However, while education is intended to align with pharmacists’ expected level of professional competencies, there is a lack of national standardized outcome-based competency criteria for new graduates and wide-ranging inter-program variations were evident. Conclusion A lack of standardized content in pharmacy qualifying programs that underpin pharmacists’ mental health knowledge and skills might result in variations to practice competencies. Further work is needed to determine how variations impact the way pharmacists deliver care to mental health consumers and their caregivers.

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

Docetaxel was the first chemotherapeutic agent to increase survival time in patients with androgen-resistant prostate cancer. However, it provides only a modest increase in survival and is associated with significant toxicity. Therefore, there is an urgent need to identify potential adjunct therapies. Given the key role of autophagy in both tumour survival and chemoresistance, the impact of autophagy modulation on docetaxel toxicity was tested in vitro. PC-3 and LNCaP cells were pre-treated with the autophagy inhibitor 3-methyladenine (5 mM) and then exposed to various concentrations (0-100 μM) of docetaxel. Cytoxic effects of docetaxel were measured using resazurin reduction to resorufin, whilst autophagy and apoptosis was measured using monodansylcadaverine, annexin V and caspase-3, respectively. Docetaxel produced significant toxicity in PC-3 cells but was not toxic to LNCaP cells. Pre-treatment with the autophagy inhibitor, 3-methyladenine (5 mM) significantly protected PC-3 cells against docetaxel-induced cytotoxicity, increased autophagosome formation and apoptosis measured using monodansylcadaverine, annexin V and caspase-3 fluorescence, respectively. In contrast, 3-methyladenine was toxic by itself in LNCaP cells and also increased autophagic vesicle formation and apoptosis but did not influence docetaxel toxicity in these cells. These paradoxical effects of 3-methyladenine were largely independent of reactive oxygen species production. We show here that modulation of autophagy may influence docetaxel-induced toxicity in prostate cancer cells and these effects may differ between cell lines.

Background Chemotherapeutic pharmacology is traditionally a challenging subject to both teach and learn. This study aimed to assess a suite of e-learning tools regarding the mechanisms of action of chemotherapy drugs, and the subsequent effects on learning in two cohorts (control and intervention) of undergraduate pharmacy students in Australia. Methods The intervention group had access to the e-learning tools in addition to the traditional didactic teaching delivered to the control group. To determine the educational benefit of the e-learning tools, we evaluated student level of understanding demonstrated in the short-answer questions related to chemotherapy mechanisms. Structure of the Observed Learning Outcome (SOLO) taxonomy was used to classify students’ exam responses. Results There was overall improvement in student total exam scores for the intervention group. However, the most significant improvement was evident in the chemotherapy scores for the intervention group who demonstrated a superior level of understanding of mechanisms of action of the three chemotherapy drugs, cytarabine, mitomycin C and trastuzumab. Students in the intervention group performed significantly better than the control cohort with respect to short answer questions on cytarabine and trastuzumab (p Discussion E-learning tools were shown to improve student level of understanding, as scored by the SOLO taxonomy, in the learning and teaching of chemotherapeutic pharmacology.

Introduction Warfarin is widely used for patients with non-valvular atrial fibrillation (NVAF). Variations in warfarin control, as measured by time in therapeutic range (TTR), have been reported across different regions and ethnicities, particularly between Western and Asian countries. However, there is limited data on comparative factors influencing warfarin control in Caucasian and Asian patients. Therefore, the aim of this study was to determine warfarin control and potential factors influencing this in patients with NVAF in Australia and Singapore. Methods Retrospective data was collected for patients receiving warfarin for January to June 2014 in Australia and Singapore. TTR was calculated for individuals with mean patient TTR used for analysis. Possible influential factors on TTR were analysed including age, gender, concurrent co-morbidities, and concurrent medication. Results The mean TTR was significantly higher in Australia (82%) than Singapore (58%). At both sites, chronic kidney disease significantly lowered this TTR. Further factors influencing control were anaemia and age Discussion Warfarin control was significantly higher in Australia compared to Singapore, however chronic kidney disease reduced control at both sites. The different levels of control in these two countries, together with patient factors further reducing control may impact on anticoagulant choice in these countries with better outcomes from warfarin in Australia compared to Singapore.

Warfarin reduces stroke risk in atrial fibrillation (AF) patients. The quality of warfarin control, measured by time in therapeutic range (TTR), impacts outcome and adverse events. One tool evaluating risk of adverse events and potential warfarin control would simplify risk-benefit assessment of warfarin. Recently, HASBLED was demonstrated effective for this purpose, but this was in well-controlled patients with deep vein thrombosis. HASBLED as a predictor of warfarin control has not been validated in other populations including differing indications, warfarin control levels and ethnicities. The aim of this study was to determine whether HASBLED can predict warfarin control in patients with AF in Australia and Singapore. Retrospective data were collected for patients receiving warfarin between January and June 2014 in Australia and Singapore. Patient data were used to calculate HASBLED at the start and end of the study period. TTR was calculated for each patient, and mean TTR used for analysis to stratified HASBLED scores. Of the 4370 patients, there were 3199 in Australia and 1171 in Singapore with mean TTRs of 82% and 58%, respectively. At the start of the study, a HASBLED score ≥3 predicted significantly lower TTR in Singapore, whilst at the end of the study, this score identified patients with poor control in both Australia and Singapore. A HASBLED score ≥3 in patients treated with warfarin can differentiate significantly lower TTRs in Australian and Singapore patients with AF. HASBLED may assess bleed risk and warfarin control, identifying patients at high risk of poor warfarin outcome requiring additional INR monitoring or alternative anticoagulation.

The anti-inflammatory effects of statins (HMG-CoA reductase inhibitors) within the cardiovascular system are well-established; however, their neuroinflammatory potential is unclear. It is currently unknown whether statins’ neurological effects are lipid-dependent or due to pleiotropic mechanisms. Therefore, the assumption that all statin compounds will have the same effect within the central nervous system is potentially inappropriate, with no studies to date having compared all statins in a single model. Thus, the aim of this study was to compare the effects of the six statins (atorvastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin) within a single in vitro model of neuroinflammation. To achieve this, PMA-differentiated THP-1 cells were used as surrogate microglial cells, and LPS was used to induce inflammatory conditions. Here, we show that pretreatment with all statins was able to significantly reduce LPS-induced interleukin (IL)-1βand tumour necrosis factor (TNF)-αrelease, as well as decrease LPS-induced prostaglandin E2 (PGE2). Similarly, global reactive oxygen species (ROS) and nitric oxide (NO) production were decreased following pretreatment with all statins. Based on these findings, it is suggested that more complex cellular models should be considered to further compare individual statin compounds, including translation into in vivo models of acute and/or chronic neuroinflammation.

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, commonly referred to as statins, are widely used in the treatment of dyslipidaemia, in addition to providing primary and secondary prevention against cardiovascular disease and stroke. Statins’ effects on the central nervous system (CNS), particularly on cognition and neurological disorders such as stroke and multiple sclerosis, have received increasing attention in recent years, both within the scientific community and in the media. Current understanding of statins’ effects is limited by a lack of mechanism-based studies, as well as the assumption that all statins have the same pharmacological effect in the central nervous system. This review aims to provide an updated discussion on the molecular mechanisms contributing to statins’ possible effects on cognitive function, neurodegenerative disease, and various neurological disorders such as stroke, epilepsy, depression and CNS cancers. Additionally, the pharmacokinetic differences between statins and how these may result in statin-specific neurological effects are also discussed.

Purpose – The purpose of this paper is to explore the potential approaches to continuing education and training delivery for community pharmacists to equip them to support mental health consumers and carers with illnesses such as depression and anxiety. Design/methodology/approach – Review of national and international literature about community pharmacists’ roles, beliefs and attitudes towards mental health, continuing education delivery for the workforce and training recommendations to equip pharmacy workforce. Findings – Training involving consumer educators was effective in reducing stigma and negative attitudes. Interactive and contextually relevant training appeared to be more effective than didactic strategies. Narratives and role-plays (from the perspective of consumers, carers and health professionals) are effective in promoting more positive attitudes and reduce stigma. Flexible on-line delivery methods with video footage of expert and consumer narratives were preferable for a cost-effective programme accessible to a wide community pharmacy workforce. Originality/value – There is a clear need for mental health education for community pharmacists and support staff in Australia. Training should target reducing stigma and negative attitudes, improving knowledge and building confidence and skills to improve pharmacy staff's perceived value of working with mental health consumers. The delivery mode should maximise uptake.

The effects of pseudomonal virulence factor pyocyanin, and LPS from Pseudomonas aeruginosa and Escherichia coli on urothelial mediator release and cytokine production were examined. RT4 urothelial cells were treated with pyocyanin (1–100 μM) or LPS (1–100 ng/mL) for 24-h. Effects were measured in terms of changes in cell viability, basal and stretch-induced acetylcholine (Ach) and PGE2 release, and inflammatory cytokines (IL-6 and IL-12) production. Twenty-four hour pyocyanin (100 μM) treatment significantly decreased urothelial cell viability, while stretch-induced Ach release response was inhibited. E. coli LPS (100 ng/mL) produced a similar response with an additional significant increase in basal Ach release. All three virulence factors significantly increased urothelial PGE2 release; under basal release for pyocyanin (100 μM), stretch-induced release for pseudomonal LPS (⩾10 ng/mL) and both basal and stimulated release for E. coli LPS (⩾10 ng/mL). IL-6 and IL-12 were not detected in control samples, however 24 h treatment with pyocyanin (100 μM) or LPS (100 ng/mL) resulted in IL-6 release from urothelial cells. The changes in urothelial Ach and PGE2, and release of inflammatory cytokine IL-6 induced by exposure to the bacterial virulence factors may play a role in the symptoms of pain and urinary urgency experienced with urinary tract infections.

The role of autophagy in pyocyanin (PCN)-induced toxicity in the central nervous system (CNS) remains unclear, with only evidence from our group identifying it as a mechanism underlying toxicity in 1321N1 astrocytoma cells. Therefore, the aim of this study was to further examine the role of autophagy in PCN-induced toxicity in the CNS. To achieve this, we exposed 1321N1 astrocytoma and SH-SY5Y neuroblastoma cells to PCN (0-100 μmol/L) and tested the contribution of autophagy by measuring the impact of the autophagy inhibitor 3-methyladenine (3-MA) using a series of biochemical and molecular markers. Pretreatment of 1321N1 astrocytoma cells with 3-MA (5 mmol/L) decreased the PCN-induced acidic vesicular organelle and autophagosome formation as measured using acridine orange and green fluorescent protein-LC3 -LC3 fluorescence, respectively. Furthermore, 3-MA (5 mmol/L) significantly protected 1321N1 astrocytoma cells against PCN-induced toxicity. In contrast pretreatment with 3-MA (5 mmol/L) increased PCN-induced toxicity in SH-SY5Y neuroblastoma cells. Given the influence of autophagy in inflammatory responses, we investigated whether the observed effects in this study involved inflammatory mediators. The PCN (100 μmol/L) significantly increased the production of interleukin-8 (IL-8), prostaglandin E2 (PGE2), and leukotriene B4 (LTB4) in both cell lines. Consistent with its paradoxical role in modulating PCN-induced toxicity, 3-MA (5 mmol/L) significantly reduced the PCN-induced production of IL-8, PGE2, and LTB4 in 1321N1 astrocytoma cells but augmented their production in SH-SY5Y neuroblastoma cells. In conclusion, we show here for the first time the paradoxical role of autophagy in mediating PCN-induced toxicity in 1321N1 astrocytoma and SH-SY5Y neuroblastoma cells and provide novel evidence that these actions may be mediated by effects on IL-8, PGE2, and LTB4 production.

Modulating gut bacteria via regular prebiotics/probiotics consumption may improve the metabolism of acute alcohol ingestion. This study investigated the impact of 8-weeks prebiotics/probiotics supplementation on microbiome changes and responses to acute alcohol consumption.38 participants (21 females, 23.6 ± 3.4 kg mIncreased counts for at least one bacterial strain (L. acidophilus or B. lactis) were observed for all participants on SYN (n = 10) and PRO (n = 10) trials. No difference in CConsumption of probiotics appears to alter faecal counts of supplemental bacterial strains in otherwise healthy individuals. However, translation to any possible beneficial impact on alcohol metabolism remains to be elucidated.

Background: Warfarin is a leading anticoagulant in the management of atrial fibrillation (AF) and deep vein thrombosis (DVT). Drug interactions influence the safety of warfarin use and while extensive literature exists regarding the effect on warfarin control and bleeding incidence with many medicines, there is little evidence on the influence of complementary medicines. The aim of this study was to assess the influence of fish and krill oil supplementation on warfarin control and bleeding incidence in AF and DVT patients. Methods: A retrospective analysis was conducted utilising patient information from a large private pathology clinic. AF and DVT patients receiving long-term warfarin therapy (>30 days) at the clinic and taking fish and krill oil supplements were eligible for study inclusion. Results: Of the 2081 patients assessed, a total of 573 warfarin users met the inclusion criteria with 145 patients in the fish and krill oil group (supplement group) and 428 patients in the control group. Overall, it was found that fish and krill oils did not significantly alter warfarin time in therapeutic range (TTR) or bleeding incidence, even when compared by gender. Conclusion: Omega-3 supplementation with fish and krill oil does not significantly affect long-term warfarin control and bleeding and thromboembolic events when consumed concurrently in patients managed at an anticoagulation clinic.

Warfarin reduces stroke risk in atrial fibrillation (AF) patients but requires ongoing monitoring. Time in therapeutic range (TTR) is used as a measure of warfarin control, with a TTR less than 60% associated with adverse patient outcomes. The Sex, Age, Medical history, Treatment, Tobacco use, Race (SAMe-TTRetrospective data were collected from the National Heart Centre Singapore for AF patients receiving warfarin between January and June 2014. The TTR and the SAMe-TTThe 1137 non-valvular AF patients had a mean TTR of 58.0 ± 34.3% and a median SAMe-TTThe SAMe-TT

This study investigated the influence of exercise-induced dehydration on alcohol pharmacokinetics, subjective ratings of impairment, and risk-taking behaviours. Twelve male volunteers participated in 3 experimental trials completed in a randomised cross over design separated by at least 7 days. In one trial, participants exercised to cause dehydration of ~ 2.5% body weight loss. For the other trials, participants were required to be in a rested and euhydrated state. A set volume of alcohol was then consumed in each trial and participants were monitored over a 4 h period. Blood (BAC) and breath (BrAC) alcohol samples were collected throughout and analysed to calculate pharmacokinetic variables associated with the blood alcohol curve. Total urine production, estimates of BrAC, and subjective ratings of intoxication and impairment were also recorded throughout each trial. No difference was found in the pharmacokinetics of alcohol between any of the trial conditions. BrACs were higher than BACs for 2 h following alcohol consumption, but lower at measures taken 3 and 4 h post ingestion. Participants' ratings of confusion and intoxication were significantly lower, and they were more willing to drive in the dehydration trial compared with one of the euhydration trials. These findings suggest that dehydration or other physiological changes associated with exercise may have an ability to influence the subjective effects of alcohol and increase the likelihood of risk-taking behaviours such as drink-driving. However, further research is required to examine the effects of alcohol under conditions of exercise-induced fluid loss in order to clarify these findings.

The purpose of the current study was to determine whether glycyrrhizin (GL) maintains hepatic glutathione (GSH) levels by inhibiting GSH biliary secretion in normal rats. The effects of glycyrrhizin on hepatic glutathione content, bile flow and biliary secretion of glutathione were examined. Because glutathione is a substrate for multidrug resistance associated protein-2 (Mrp2/ABCC2), the inhibitory effects of GL on Mrp2 in isolated perfused rat liver and in Mrp2-expressing Sf9 membrane vesicles were also examined using the Mrp2 substrate methotrexate (MTX) and estradiol-17-β-glucuronide (E217G). The hepatic content of glutathione in rats following GL perfusion (43.7 µmol/l) in isolated liver perfusion and GL intravenous treatment (25 mg/kg) was significantly higher than that for the control. A marked and dose-dependent decrease in the excretion of glutathione was observed. In addition, the secretion rate of MTX was decreased by 57% in isolated liver perfusion in GL-treated rats. Moreover the ATP-dependent uptake of E217G by Mrp2 membrane vesicles was decreased by 75.9% in the 20 µm GL group and by 60.5% in the 2 µm GL group. In conclusion, glycyrrhizin increases hepatic glutathione content possibly through inhibition of Mrp2 which then reduces the biliary excretion of glutathione. Copyright © 2012 John Wiley & Sons, Ltd.

Central nervous system (CNS) infections due to Pseudomonas aeruginosa are difficult to treat and have a high mortality rate. Pyocyanin, a virulence factor produced by P. aeruginosa, has been shown to be responsible for the majority of P. aeruginosa's pathogenicity in mammalian cells. Several lines of evidence in respiratory cells suggest that this damage is primarily mediated by pyocyanin's ability to generate ROS and deplete host antioxidant defense mechanisms. However, it has yet to be established whether pyocyanin or 1-hydroxyphenazine have potential toxicity to the CNS. Therefore, the aim of this study was to compare the CNS toxicity of pyocyanin and 1-hydroxyphenazine in vitro and to provide insight into mechanisms that underlie this toxicity using 1321N1 astrocytoma cells. To achieve this, we investigated the contribution of oxidative stress and other mediators of cell death including autophagy, senescence and apoptosis. We show that oxidative stress is not a primary mediator of pyocyanin (0-100 μM) and 1-hydroxyphenazine (0-100 μM) induced toxicity in 1321N1 cells. Instead, our results suggest that autophagy may play a central role. The autophagy inhibitor 3-methyladenine (5 mM) protected 1321N1 astrocytoma cells against both pyocyanin and 1-hydroxyphenazine-induced cell injury and increased accumulation of acidic vesicular organelles, a hallmark of autophagy. Furthermore, apoptosis and senescence events may be secondary to autophagy in pyocyanin and 1-hydroxyphenazine-mediated cell injury. In conclusion, this study provides the first evidence on mechanisms underlying the toxicity of both pyocyanin and 1-hydroxyphenazine to astrocytoma cells and provides novel evidence suggesting that this toxicity may be mediated by the formation of acidic vesicular organelles, a hallmark of autophagic cell death.

This paper describes the development of a sensitive high performance liquid chromatography (HPLC) method for quantification of rosiglitazone in sheep plasma and amniotic fluid. Samples were prepared by liquid-liquid extraction using tert-butyl methyl ether, and rosiglitazone was quantitated by HPLC using a C18 column and fluorescence detector with an excitation wavelength of 247 nm and emission wavelength of 367 nm. The mobile phase consisted of ammonium acetate (10 mM, pH 5.2) and acetonitrile (56.5:43.5, v/v) with a flow rate of 1 ml/min. Ketoconazole was used as the internal standard (IS). The plasma calibration curve was linear over the range of 2.5-250 ng/ml (mean r2 = 0.9940 ± 0.0024; n = 6) with accuracy of 99.4-102.8% over the calibration range. The intra-day and inter-day coefficient of variation (%CV, percent coefficient of variation) were in the range of 0.01-8.68% in sheep plasma. Similar performance was achieved for amniotic fluid. The described method was successfully applied to quantitate rosiglitazone concentrations in the pregnant ewe and her fetus. Refereed/Peer-reviewed

The aim of the study was to investigate the effects of baicalin on blood glucose, insulin and cytokine levels. Rat diabetes was induced by intraperitoneal (i.p.) injection of nicotinamide and streptozotocin. Diabetic rats were dosed with i.p. baicalin or oral metformin daily for 8 days. Blood glucose, insulin and hepatic glycogen were determined using conventional methods. The activity of hepatic hexokinase was determined using a coupled assay with glucose-6-phosphate dehydrogenase. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and adiponectin were measured by enzyme-linked immunosorbent assay. Administration of baicalin at 50 or 100 mg/kg significantly decreased plasma glucose levels in a dose dependent manner. The serum insulin level was not increased by baicalin treatment. Administration of baicalin at a high dose (100 mg/kg) resulted in a significant increase of liver glycogen content and a reduction of serum TNF-α. The activity of hepatic hexokinase was significantly increased after dosing baicalin at 25, 50 or 10 mg/kg. Administration of baicalin (50 or 10 mg/kg) or metformin (10 mg/kg) significantly alleviated the morphological injury to the pancreas caused by STZ. The possible mechanisms contributing to the hypoglycemic effect include increasing the hepatic glycogen content and glycolysis, and reducing the serum levels of TNF-α. Copyright © 2010 John Wiley & Sons, Ltd.

The aim of present study was to investigate the effect of baicalin on hypoxia/reoxygenation (H/R) injury in cardiomyocytes and the mechanisms involved, particularly in relation to cytokines. The cardiomyocytes for the H/R groups were placed into a hypoxic chamber for 12 h and then underwent reoxygenation for 1 h. The cells in the TNF-alpha groups were administered 100 ng/mL rrTNF-alpha and incubated for 13 h under normal conditions. The cells in the baicalin pretreatment groups were administered 10 microM baicalin 30 min prior to exposure to H/R or TNF-alpha. It was observed that pretreatment with baicalin (10 microM) significantly reduced the cell damage and death induced by H/R or TNF-alpha. In the culture medium of the H/R cells, the SOD activity increased, while TNF-alpha was decreased by baicalin. The levels of IL-6 in culture medium for H/R or TNF-alpha treated cells were suppressed by baicalin pretreatment. In contrast, the levels of IL-10 in culture medium for H/R or TNF-alpha treated cells were significantly elevated by baicalin. Moreover, baicalin inhibited the nuclear translocation of NF-kappaB induced by H/R or TNF-alpha. In conclusion, baicalin may protect cardiomyocytes from H/R injury through an anti-inflammatory mechanism.

To study the metabolites of glycyrrhetic acid (GA) in rat bile, an analytical method was developed to identify GA and its metabolites by liquid chromatography mass spectrometry (LC-MS). Rat bile was collected after i.v. injection of GA. Three major GA-related peaks were detectable in rat bile by high-performance liquid chromatography (HPLC) analysis at 254 nm. LC-MS spectra showed their protonated molecular ions at m/z 727, 647, and 551. Furthermore, the three metabolites were also confirmed to exist in rat bile on LC-MS total ion chromatogram (TIC). Taken together with the susceptible nature to beta-glucuronidase digestion and alkaline conditions, they were identified to be a novel sulfate-glucuronide diconjugate and the known monoglucuronide and sulfate conjugate, respectively.

Despite the fact that fetal drug exposure is common, the disposition of drugs in the fetus is poorly understood.This study aimed to investigate fetal placental and non-placental disposition of rosiglitazone inthe pregnant ewe. Steady state was reached after day 5 of fetal infusion, and were ∼1.8 fold higherthan maternal concentrations (P < 0.001). The AUC for fetal rosiglitazone concentration throughout theinfusion was inversely correlated with placental and fetal weight. Metabolic activity of the fetal livermicrosomes were ∼25 fold lower than maternal microsomes (P < 0.001). The findings suggest that transplacentaltransfer is the major route through which rosiglitazone is cleared from the fetal compartment,while non-placental hepatic elimination makes only a minor contribution. This supports a limited capacityof the fetus for eliminating this class of drugs, and highlights the potential for drug toxicity whenadministering pharmacotherapy to the mother/fetus in human pregnancy. Refereed/Peer-reviewed

Damage to the peritoneum during abdominal surgery triggers a cascade of events aimed at repairing the damage. As part of this process, fibrin is deposited, which is the precursor to the formation of an adhesion between 2 damaged peritoneal surfaces. This can have a significant impact on morbidity and even mortality as well as large cost implications. Strategies to reduce adhesion formation include improving surgical techniques, optimizing laparoscopy conditions, using pharmacologic interventions targeted at the inflammatory response and/or fibrin deposition, and using agents that provide a physical barrier to adhesion formation. While these strategies have provided some success, none have yet proved totally successful in abolishing adhesions. Further research to ensure that adhesion prevention is optimal is therefore essential.