Impact of a pharmacy-led smoking cessation clinic in a dermatology centre.

INTRODUCTION: Cigarette smoking is a leading cause of morbidity and mortality, and has a deleterious effect on dermatological conditions, such as skin cancers, hidradenitis suppurativa and psoriasis. The study aimed to evaluate the efficacy of a pharmacist-led smoking cessation clinic in reducing cigarette smoking at a tertiary referral dermatology centre. We described the impact of this clinic to provide guidance on how such a model could be further improved and implemented more widely. METHODS: In this single-centre, retrospective study, 74 currently smoking patients who received counselling at a structured smoking cessation clinic between January 2010 and March 2013 were identified. Information on baseline demographic characteristics and detailed past medical history, including smoking history, was collected. Follow-up was conducted at two weeks and three months. RESULTS: At the first follow-up at two weeks, which was attended by 57 patients, 9 (15.8%) had stopped smoking and 26 (45.6%) showed reduction in the number of cigarette sticks smoked per day, with an average reduction of 4.1 cigarette sticks per day. However, a few patients also reported no change or increased number of cigarette sticks smoked per day following counselling. CONCLUSION: A structured pharmacist-led smoking cessation clinic is effective and can be made a part of the holistic management of dermatological conditions.

Measurement of LRRK2 and Ser910/935 phosphorylated LRRK2 in peripheral blood mononuclear cells from idiopathic Parkinson's disease patients.

A significant number of autosomal dominantly inherited Parkinson's disease (PD) cases are due to mutations in the leucine-rich repeat kinase 2 (LRRK2) gene. In cells, these pathogenic mutations have a number of differing effects on LRRK2 enzymatic activity and protein stability. In particular, five of the six described pathogenic LRRK2 mutations ablate the constitutive phosphorylation of LRRK2 on Ser910 and Ser935, two residues required for binding of LRRK2 to 14-3-3 proteins. This suggests a potential pathogenic role for these residues. However, LRRK2 kinase inhibitors, which have shown early promise as neuroprotective agents, also ablate the phosphorylation of Ser910 and Ser935. Additionally, LRRK2 is phosphorylated on Ser910 and Ser935 following activation of the inflammatory toll-like receptor pathway and inflammatory cytokines are often increased in PD patients. Whether LRRK2 protein or phosphorylation is altered in idiopathic PD is unknown. We therefore measured LRRK2 protein and its phosporylation in peripheral blood mononuclear cells (PBMCs) from 33 idiopathic Parkinson's disease patients and 27 age-matched controls. We found no significant difference in total LRRK2 protein levels in PBMCs from PD patients compared to controls. Furthermore, total LRRK2 protein expression was not effected by age, disease duration, disease severity or levodopa medication. The amount of phosphorylation on LRRK2 at both Ser910 and Ser935 correlated highly with total LRRK2 levels and was also unchanged in PD patients. Therefore, changes in LRRK2 Ser910/Ser935 phosphorylation in PBMCs are unlikely to contribute to idiopathic Parkinson's disease or be of utility as a disease biomarker. However, the invariance of Ser910 and Ser935 phosphorylation in PD PBMC's suggests that these residues could be used as pharmacodynamic biomarkers for the effectiveness of LRRK2 kinase inhibitors in patients.

Macroautophagy in sporadic and the genetic form of Parkinson's disease with the A53T α-synuclein mutation.

BACKGROUND: The A53T mutation in the α-synuclein gene causes autosomal-dominant Lewy body Parkinson's disease (PD). Cultured cell models have linked this mutation to increased cell macroautophagy, although evidence of enhanced macroautophagy in patients with this mutation has not been assessed. OBJECTIVE: To determine whether macroautophagy is increased by the A53T α-synuclein gene mutation in PD patients and cell models. METHODS: Formalin-fixed paraffin-embedded 10 µm-thick tissue sections from the substantia nigra and anterior cingulate cortex of two PD patients with the A53T α-synuclein gene mutation were compared with four sporadic PD cases and four controls obtained from the Sydney Brain Bank. Lewy bodies were isolated from frontal cortex of a case with late stage PD (recruited from South Australian Brain Bank). Immunohistochemistry was performed for α-synuclein and the macroautophagy markers autophagy-specific gene (ATG) 5, ATG6/Beclin1 and ATG8/LC3. SH-SY5Y cells were transfected with wild type or A53T mutant α-synuclein plasmids and observable changes in macroautophagy marker protein levels assessed using Western blotting. RESULTS: α-Synuclein immunoreactive neurites and dots were more numerous in patients with A53T mutations compared with late stage sporadic PD patients, and perinuclear cytoplasmic α-synuclein aggregates were observed in the α-synuclein A53T gene transfected SH-SY5Y cells compared to wild type transfections. All PD patients (with or without A53T mutations) had increased immunohistochemical evidence for macroautophagy compared with controls, and the levels of the ATG5 complex were equally increased in wild type and A53T α-synuclein gene transfected cells compared to controls. CONCLUSION: Despite increased α-synuclein accumulation with A53T mutations, macroautophagy is not increased above that observed in sporadic patients with PD or in cells transfected with wild type α-synuclein, suggesting that mutated α-synuclein protein is not removed by macroautophagy.