Oxygen limitation fails to explain upper chronic thermal limits and the temperature size rule in mayflies.

An inability to adequately meet tissue oxygen demands has been proposed as an important factor setting upper thermal limits in ectothermic invertebrates (especially aquatic species) as well as explaining the observed decline in adult size with increased rearing temperature during the immature stages (a phenomenon known as the temperature size rule, or TSR). We tested this by rearing three aquatic insects (the mayflies Neocloeon triangulifer and two species of the Cloeon dipterum complex) through their entire larval life under a range of temperature and oxygen concentrations. Hyperoxia did not extend upper thermal limits, nor did it prevent the loss of size or fertility experienced near upper chronic thermal limits. At moderate temperatures, the TSR pattern was observed under conditions of hyperoxia, normoxia and hypoxia, suggesting little or no influence of oxygen on this trend. However, for a given rearing temperature, adults were smaller and less fecund under hypoxia as a result of a lowering of growth rates. These mayflies greatly increased the size of their gills in response to lower dissolved oxygen concentrations but not under oxygen-saturated conditions over a temperature range yielding the classic TSR response. Using ommatidium diameter as a proxy for cell size, we found the classic TSR pattern observed under moderate temperature conditions was due primarily to a change in the number of cells rather than cell size. We conclude overall that a failure to meet tissue oxygen demands is not a viable hypothesis for explaining either the chronic thermal limit or TSR pattern in these species.

Design and Characterization of Injectable Poly(Lactic-Co-Glycolic Acid) Pastes for Sustained and Local Drug Release.

PURPOSE: We describe the preparation of injectable polymeric paste (IPP) formulations for local and sustained release of drugs. Furthermore, we include the characterization and possible applications of such pastes. Particular attention is paid to characteristics relevant to the successful clinical formulation development, such as viscosity, injectability, degradation, drug release, sterilization, stability performance and pharmacokinetics. METHODS: Paste injectability was characterized using measured viscosity and the Hagen-Poiseuille equation to determine injection forces. Drug degradation, release and formulation stability experiments were performed in vitro and drug levels were quantified using HPLC-UV methods. Pharmacokinetic evaluation of sustained-release lidocaine IPPs used five groups of six rats receiving increasing doses subcutaneously. An anti-cancer formulation was evaluated in a subcutaneous tumor xenograft mouse model. RESULTS: The viscosity and injectability of IPPs could be controlled by changing the polymeric composition. IPPs demonstrated good long-term stability and tunable drug-release with low systemic exposure in vivo in rats. Preliminary data in a subcutaneous tumor model points to a sustained anticancer effect. CONCLUSIONS: These IPPs are tunable platforms for local and sustained delivery of drugs and have potential for further clinical development to treat a number of diseases.

Enhanced taxane uptake into bladder tissues following co-administration with either mitomycin C, doxorubicin or gemcitabine: association to exfoliation processes.

OBJECTIVE: To investigate the effect of three anticancer drugs (mitomycin c (MMC), doxorubicin or gemcitabine) on bladder wall morphology and the uptake of paclitaxel or docetaxel following coadministration. The primary objective of this study was to measure the uptake of MMC, doxorubicin or gemcitabine with or without exposure of the tissue to amine terminated cationic nanoparticles (CNPs) and to investigate any possible exfoliation effects of the three drugs on intact bladder tissue. The secondary objective was to investigate the uptake of taxane drugs (docetaxel, DTX) and paclitaxel, (PTX) from surfactant micelle formulations in the presence of MMC, doxorubicin or gemcitabine. MATERIALS AND METHODS: Sections of fresh pig bladder tissue were incubated in Franz diffusion cells with the urothelial side exposed to solutions of doxorubicin, MMC and gemcitabine containing radioactive drug for 90 min. Some tissue samples were simultaneously exposed to each of the three drugs in combination with the surfactant micelle formulations of PTX (Taxol) or DTX (Taxotere). Tissue sections were then cryostat sectioned for drug quantitation by liquid scintillation counting or fixed for scanning electron microscopy and haematoxylin and eosin staining. RESULTS: All three drugs caused exfoliation of the urothelial layer of bladder tissues. Drug uptake studies showed that all three drugs effectively penetrated the lamina propria through to the muscular layer over a 2-h incubation and these levels were unaffected by pre-treatment with CNPs. The uptake levels of the taxane drugs PTX and DTX were significantly enhanced following simultaneous treatment of bladders with MMC, doxorubicin or gemcitabine. CONCLUSION: The exfoliation effects of MMC, doxorubicin and gemcitabine allow for good tissue penetration of these drugs with no additional effect from CNP treatment of bladders. The observed exfoliation effect of these amine-containing drugs probably arises from a cationic interaction with the mucus and urothelium cell layer in a manner similar to that previously reported for CNPs. These studies suggest that the lack of long-term clinical efficacy of these drugs may not arise from poor intravesical drug penetration but may result from a rapid diffusion of the drugs into the deeper vascularised muscular region with rapid drug clearance. The enhanced uptake of PTX or DTX following co-administration with MMC, doxorubicin or gemcitabine probably arises from the removal of the urothelial barrier by exfoliation allowing for improved taxane partitioning into superficial layers. These effects may allow for dual drug intravesical strategies offering greatly improved taxane uptake and potential additive drug effects for improved efficacy.

Magnetically-actuated drug delivery device (MADDD) for minimally invasive treatment of prostate cancer: An in vivo animal pilot study.

BACKGROUND: The vast majority of prostate cancer presents clinically localized to the prostate without evidence of metastasis. Currently, there are several modalities available to treat this particular disease. Despite radical prostatectomy demonstrating a modest prostate cancer specific mortality benefit in the PIVOT trial, several novel modalities have emerged to treat localized prostate cancer in patients that are either not eligible for surgery or that prefer an alternative approach. METHODS: Athymic nude mice were subcutaneously inoculated with prostate cancer cells. The mice were divided into four cohorts, one cohort untreated, two cohorts received docetaxel (10 mg/kg) either subcutaneously (SC) or intravenously (IV) and the fourth cohort was treated using the magnetically-actuated docetaxel delivery device (MADDD), dispensing 1.5 µg of docetaxel per 30 min treatment session. Treatment in all three therapeutic arms (SC, IV, and MADDD) was administered once weekly for 6 weeks. Treatment efficacy was measured once a week according to tumor volume using ultrasound. In addition, calipers were used to assess tumor volume. RESULTS: Animals implanted with the device demonstrated no signs of distress or discomfort, neither local nor systemic symptoms of inflammation and infection. Using an independent sample t-test, the tumor growth rate of the treated tumors was significant when compared to the control. Post hoc Tukey HSD test results showed that the mean tumor growth rate of our device cohort was significantly lower than SC and control cohorts. Moreover, IV cohort showed slight reduction in mean tumor growth rates than the ones from the device cohort, however, there was no statistical significance in tumor growth rate between these two cohorts. Furthermore, immunohistochemistry demonstrated an increased cellular apoptosis in the MADDD treated tumors and a decreased proliferation when compared to the other cohorts. In addition, IV cohort showed increased treatment side effects (weight loss) when compared to the device cohort. Finally, MADDD showed minimal expression of CD45 comparable to the control cohort, suggesting no signs of chronic inflammation. CONCLUSIONS: In conclusion, this study showed for the first time that MADDD, clearly suppressed tumor growth in local prostate cancer tumors. This could potentially be a novel clinical treatment approach for localized prostate cancer.

Physiological responses to short-term thermal stress in mayfly (Neocloeon triangulifer) larvae in relation to upper thermal limits.

Understanding species' thermal limits and their physiological determinants is critical in light of climate change and other human activities that warm freshwater ecosystems. Here, we ask whether oxygen limitation determines the chronic upper thermal limits in larvae of the mayfly Neocloeon triangulifer, an emerging model for ecological and physiological studies. Our experiments are based on a robust understanding of the upper acute (∼40°C) and chronic thermal limits of this species (>28°C, ≤30°C) derived from full life cycle rearing experiments across temperatures. We tested two related predictions derived from the hypothesis that oxygen limitation sets the chronic upper thermal limits: (1) aerobic scope declines in mayfly larvae as they approach and exceed temperatures that are chronically lethal to larvae; and (2) genes indicative of hypoxia challenge are also responsive in larvae exposed to ecologically relevant thermal limits. Neither prediction held true. We estimated aerobic scope by subtracting measurements of standard oxygen consumption rates from measurements of maximum oxygen consumption rates, the latter of which was obtained by treating with the metabolic uncoupling agent carbonyl cyanide-4-(trifluoromethoxy) pheylhydrazone (FCCP). Aerobic scope was similar in larvae held below and above chronic thermal limits. Genes indicative of oxygen limitation (LDH, EGL-9) were only upregulated under hypoxia or during exposure to temperatures beyond the chronic (and more ecologically relevant) thermal limits of this species (LDH). Our results suggest that the chronic thermal limits of this species are likely not driven by oxygen limitation, but rather are determined by other factors, e.g. bioenergetics costs. We caution against the use of short-term thermal ramping approaches to estimate critical thermal limits (CTmax) in aquatic insects because those temperatures are typically higher than those that occur in nature.

Identification of major factors in Australian primary care pharmacists' practice environment that have a bearing on the implementation of professional models of practice.

Objective The aim of the present study was to describe an environmental framework for pharmacists in primary care in Australia and determine the major factors within that environment that have the greatest bearing on their capacity to implement patient-focused models of professional practice. Methods A draft framework for pharmacists' practice was developed by allocating structures, systems and related factors known to the researchers or identified from the literature as existing within pharmacists' internal, operational and external environments to one of five domains: Social, Technological, Economic, Environmental or Political [STEEP]. Focus groups of pharmacists used an adapted nominal group technique to assess the draft and add factors where necessary. Where applicable, factors were consolidated into groups to establish a revised framework. The three major factors or groups in each domain were identified. The results were compared with the enabling factors described in the profession's vision statement. Results Seventy-eight individual factors were ultimately identified, with 86% able to be grouped. The three dominant groups in each of the five domains that had a bearing on the implementation of professional models of practice were as follows: (1) Social: the education of pharmacists, their beliefs and the capacity of the pharmacist workforce; (2) Technological: current and future practice models, technology and workplace structures; (3) Economic: funding of services, the viability of practice and operation of the Pharmaceutical Benefits Scheme; (4) Environmental: attitudes and expectations of stakeholders, including consumers, health system reform and external competition; and (5) Political: regulation of practice, representation of the profession and policies affecting practice. Conclusions The three dominant groups of factors in each of the five STEEP environmental domains, which have a bearing on pharmacists' capacity to implement patient-focused models of practice, correlate well with the enabling factors identified in the profession's vision statement, with the addition of three factors in the Environmental domain of stakeholder attitudes, health system reform and external competition. What is known about the topic? The extensive range of patient-focused professional programs developed for application by pharmacists in primary care in Australia has yet to be widely implemented. What does this paper add? Factors both within and beyond the pharmacists' immediate practice environment that have a bearing on the uptake of professional programs have been identified and prioritised using a structured thematic approach. What are the implications for practitioners? The results demonstrate the need for a multifactorial approach to the implementation of professional models of practice in this setting.

Tissue Permeability Effects Associated with the Use of Mucoadhesive Cationic Nanoformulations of Docetaxel in the Bladder.

PURPOSE: Recently, efficacy studies in mice have shown that amine-terminated cationic (CNP) nanoparticulate carriers of DTX offer an improved formulation of the drug for intravesical delivery. It is hypothesized that this improved efficacy may arise from a carrier mediated bladder exfoliation process that removes the urothelial barrier allowing for increased drug uptake into bladder tissue. The objective of this study was to investigate exfoliation processes in fresh pig's bladders (ex vivo) exposed to three cationic polyglycerols with increasing degrees of amination (denoted 350, 580 and 780). The study also compared the tissue depth profile of DTX uptake into these tissues using these different carriers. MATERIALS AND METHODS: Aminated polyglycerols were synthesized and characterized in the laboratory with low (CNP-360), medium (CNP-580) and high (CNP-780) levels of amine content. CNP-based DTX solutions and commercial DTX solutions in polysorbate 80 (Taxotere®) were doped with (3)H-radiolabeled DTX and prepared by solvent evaporation from acetonitrile, followed by drying and reconstitution in pH 6.4 buffer. Sections of fresh pig's bladder tissue were clamped into Franz diffusion cells and the urothelial side was exposed to the DTX solutions for 2 h. Tissue sections were then frozen for sectioning by cryotome sectioning and subsequently processed for drug analysis by liquid scintillation counting. Alternatively tissue sections were fixed in 2% glutaraldehyde and 2% paraformaldehyde in 0.1 M sodium cacodylate buffer for the purposes of scanning electron microscopy (SEM). RESULTS: Exposure of the urothelial surface to the amine-terminated polyglycerol solutions resulted in the exfoliation of bladder tissues in a time- and concentration-dependent manner. Exfoliation was significantly more pronounced when using CNPs with a medium or high levels of amination whereas only minor levels of exfoliation were seen with low levels. Following incubation of tissues in Tween-based commercial formulations (Taxotere) of DTX (0.5 mg/mL) the drug was detectable at low levels (10-40 µg/g tissue) in all depths of tissue. Similar drug uptake was observed using the CNP-360 formulation. However drug uptake levels were increased to 60-100 µg/g tissue when samples were incubated with either the CNP-580 or CNP-780 formulations. CONCLUSION: The use of cationic polyglycerols with higher levels of amine termination allows for an enhanced uptake of DTX into bladder tissues as compared to commercial (Taxotere) formulations. These increased drug levels probably arise from exfoliation processes resulting in a temporary elimination of the urothelial permeability barrier and increased drug penetration into the tissue.

The use of tissue sealants to deliver antibiotics to an orthopaedic surgical site with a titanium implant.

BACKGROUND: Orthopaedic surgery is associated with unacceptable infection rates that respond poorly to systemic antibiotics. The objective of this study was to use an animal model for orthopaedic implant infection to examine the ability of a new-generation fibrin tissue sealant to effectively deliver antibiotics to the surgical site. METHODS: The antibiotics cefazolin, fusidic acid or 5-fluorouracil were blended into Vitagel tissue sealant. The release rate of the drugs was measured using HPLC methods and bioactivity was measured by the zone of inhibition method with pathogenic Staphylococcus aureus. The antibiotic activity of the drug-loaded sealant was then tested in rats using infected orthopaedic surgical sites (titanium clip on spine). Efficacy was evaluated by residual bacterial counts on clips, clinical observations of infection, and histological findings. RESULTS: The drugs were released in a controlled manner over 2-4 days. All three antibiotics demonstrated strong antibacterial activity when released from the sealants. None of the treated animals demonstrated systemic illness. Post mortem dissection revealed a well-encapsulated abscess surrounding the titanium clip with erosion of the bony process. Using an inoculum of 1-5 × 10(3) CFU, treatment with antibiotic-loaded fibrin sealant demonstrated reduced infective swelling and reduced bacterial counts on surgical clip swabs compared to control rats or rats treated with antibiotic only. This model allowed for almost 100 % infectivity with a 0 % mortality rate due to infection, mimicking the clinical features of human implant infection. CONCLUSION: The results support the use of antibiotic-loaded commercially available fibrin sealants to prevent infection after implant surgery.

An evaluation of the Australian Community Pharmacy Agreement from a public policy perspective: industry policy cloaked as health policy?

BACKGROUND: A series of Community Pharmacy Agreements (Agreements) between the Federal government and a pharmacy-owners' body, the Pharmacy Guild of Australia (PGA) have been influential policy in Australian community pharmacy (CP) since 1990. While ostensibly to support the public's access and use of medicines, the core elements of the Agreements have been remuneration for dispensing and rules that limit the establishment of new pharmacies. Criticism has focused on the self-interest of pharmacy owners, the exclusion of other pharmacy stakeholders from the Agreement negotiations, the lack of transparency, and the impact on competition. The objective of this paper is to determine the true nature of the policy by examining the evolution of the CPA from a policy theory perspective. METHODS: A qualitative evaluation of all seven Agreement documents and their impact was undertaken using policy theories including a linear policy development model, Multiple Streams Framework, Incremental Theory, the Advocacy Coalition Framework, the Theory of Economic Regulation, the Punctuated Equilibrium Framework, and Elite Theory. The Agreements were evaluated using four lenses: their objectives, evidentiary base, stakeholders and beneficiaries. RESULTS: The PGA has acted as an elite organisation with long-standing influence on the policy's development and implementation. Notable has been the failure of other pharmacy stakeholders to establish broad-based advocacy coalitions in order to influence the Agreements. The incremental changes negotiated every 5 years to the core elements of the Agreements have supported the publics' access to medication, provided stability for the government, and security for existing pharmacy owners. Their impact on the evolution of pharmacists' scope of practice and through that, on the public's safe and appropriate use of medication, has been less clear. CONCLUSIONS: The Agreements can be characterised predominantly as industry policy benefiting pharmacy owners, rather than health policy. An emerging issue is whether incremental change will continue to be an adequate policy response to the social, political, and technological changes that are affecting health care, or whether policy disruption is likely to arise.

A Qualitative Evaluation of the Australian Community Pharmacy Agreement.

The Australian Federal Government's Community Pharmacy Agreement (Agreement), initiated in 1990 and renegotiated every five years with a pharmacy owners' organisation, is the dominant policy directing community pharmacy. We studied the experience with the Agreements of 38 purposively selected individual pharmacists and others of diverse backgrounds, using in-depth, semi-structured interviews. Although perceived to lack transparency in negotiation and operation, as well as paucity of outcome measures, the Agreements have generally supported the viability of community pharmacies and on balance, contributed positively to the public's access to medicines. There were, however, contradictory opinions regarding the impact of the policy's regulation of pharmacy locations, including the suggestion that they provide existing owners with an undue commercial advantage. A reported shortcoming of the Agreements was their impact on pharmacists' abilities to expand their scopes of practice and assist patients to make better use of medicines, in part due to the funding being almost totally focused on supply-related functions. The support for programs such as medication management services was perceived to be limited, and opportunities for diversification in pharmacy practice appeared constrained. Future pharmacy policy developed by the government could be more inclusive of a diverse range of stakeholders, seek to better utilise pharmacists' expertise, and have a greater focus on health outcomes.

Abandoned coal mine drainage and its remediation: impacts on stream ecosystem structure and function.

The effects of abandoned mine drainage (AMD) on streams and responses to remediation efforts were studied using three streams (AMD-impacted, remediated, reference) in both the anthracite and the bituminous coal mining regions of Pennsylvania (USA). Response variables included ecosystem function as well as water chemistry and macroinvertebrate community composition. The bituminous AMD stream was extremely acidic with high dissolved metals concentrations, a prolific mid-summer growth of the filamentous alga, Mougeotia, and > 10-fold more chlorophyll than the reference stream. The anthracite AMD stream had a higher pH, substrata coated with iron hydroxide(s), and negligible chlorophyll. Macroinvertebrate communities in the AMD streams were different from the reference streams, the remediated streams, and each other. Relative to the reference stream, the AMD stream(s) had (1) greater gross primary productivity (GPP) in the bituminous region and undetectable GPP in the anthracite region, (2) greater ecosystem respiration in both regions, (3) greatly reduced ammonium uptake and nitrification in both regions, (4) lower nitrate uptake in the bituminous (but not the anthracite) region, (5) more rapid phosphorus removal from the water column in both regions, (6) activities of phosphorus-acquiring, nitrogen-acquiring, and hydrolytic-carbon-acquiring enzymes that indicated extreme phosphorus limitation in both regions, and (7) slower oak and maple leaf decomposition in the bituminous region and slower oak decomposition in the anthracite region. Remediation brought chlorophyll concentrations and GPP nearer to values for respective reference streams, depressed ecosystem respiration, restored ammonium uptake, and partially restored nitrification in the bituminous (but not the anthracite) region, reduced nitrate uptake to an undetectable level, restored phosphorus uptake to near normal rates, and brought enzyme activities more in line with the reference stream in the bituminous (but not the anthracite) region. Denitrification was not detected in any stream. Water chemistry and macroinvertebrate community structure analyses capture the impact of AMD at the local reach scale, but functional measures revealed that AMD has ramifications that can cascade to downstream reaches and perhaps to receiving estuaries.

The application of layered double hydroxide clay (LDH)-poly(lactide-co-glycolic acid) (PLGA) film composites for the controlled release of antibiotics.

Many sites of bacterial infection such as in-dwelling catheters and orthopedic surgical sites require local rather than systemic antibiotic administration. However, currently used controlled release vehicles, such as polymeric films, release water-soluble antibiotics too quickly, whereas nonporous bone cement, used in orthopedics, release very little drug. The purpose of this study was to investigate the use of nanoparticulates composed of layered double hydroxide clays to bind various antibiotics and release them in a controlled manner. Mg-Al (carbonate) layered double hydroxides were synthesized and characterized using established methods. These clay particles were suspended in solutions of the antibiotics tetracycline, doxorubicin (DOX), 5-fluorouracil, vancomycin (VAN), sodium fusidate (SF) and antisense oligonucleotides and binding was determined following centrifugation and quantitation of the unbound fraction by UV/Vis absorbance or HPLC analysis. Drug release from layered double hydroxide clay/drug complexes dispersed in polymeric films was measured by incubation in phosphate-buffered saline (pH 7.4) at 37 °C using absorbance or HPLC analysis. Antimicrobial activity of drug released from film composites was determined using zonal inhibition studies against S. epidermidis. All drugs bound to the clay particles to various degrees. Generally, drugs released with a large burst phase of release (except DOX) with little further drug release after 4 days. Dispersion of drug/clay complexes in poly(lactic-co-glycolic acid) films resulted in a reduced burst phase of release and a slow continuous release for many weeks with effective antimicrobial amounts of VAN and SF released at later time points. Layered double hydroxide clays may be useful for controlled release applications at sites requiring long-term antibiotic exposure as they maintain the drug in a non-degraded state and release effective amounts of drug over long time periods. LDH clay/drug complexes are amenable to homogenous dispersion in polymeric films where implant coating may be optimal or required.

Controlled and localized antibiotics delivery using magnetic-responsive beads for synergistic treatment of orthopedic infection.

Antibiotic-loaded bone cement beads have been a reliable passive delivery system for the localized treatment of osteomyelitis; however, low, and unregulated drug release rates limit the ability of this system to maintain therapeutic concentrations. This problem is further amplified by drug-resistant pathogens that might invade or evolve under these conditions. Furthermore, currently available bone cements are incompatible with some antibiotics. The proposed device resembles conventional bone cement beads but contains an on-demand drug delivery magnetic sponge that provides actively controlled release of antibiotics. The slightly porous structure facilitates some drug diffusion while further drug release may be controlled remotely via magnetic actuation. Additionally, a combination of silver nitrate and gentamicin are used in the device as these agents are shown to display a synergistic antibacterial activity in vitro using checkerboard and time-kill assays. The device releases gentamicin and silver in both actuation and diffusion modes over 7 days. The in vitro bacterial studies demonstrate the efficacy of the released agents alone, and synergistically in combination, against Methicillin-resistant Staphylococcus aureus and Escherichia coli. The proposed device offers a facile fabrication process which allows control of the release profile by engineering hole configurations or manipulating magnetic field strength to provide the most effective therapy.

Characterization of polymethylmethacrylate microspheres loaded with silver and doxycycline for dental materials applications.

OBJECTIVES: The manufacturing of polymethylmethacrylate(PMMA) microspheres loaded with doxycycline(DOX) and/or silver sulfate(Ag2SO4) to be incorporated into glass ionomer cement(GIC). METHODS: PMMA microspheres were manufactured with Ag2SO4(1-5%) and/or DOX(5-15%). Particle size, encapsulation efficiency and drug release were measured by light microscope, ICP, and HPLC. Microspheres were added to a dental GIC(20%w/w). Drug release and DTS were investigated. Minimum inhibitory concentration and antibacterial effects of PMMA microspheres into GIC materials were tested. RESULTS: The median diameter of 50 µm was obtained for microspheres. DOX was encapsulated at an efficiency of 8.3% using a theoretical loading of 15%DOX + 5%Ag2SO4. The Ag2SO4 encapsulation efficiency was 0.63% using a theoretical loading of 5%AgSO4. All groups showed burst release within the first day and continued released up to 15 days, with 60-83% of DOX and approximately 30% of silver. For GIC, approximately 15% of DOX and 0.18% of silver were released in a 7-day period. Microbiological results showed an antimicrobial effect against S. mutans when the lead formulation of microspheres was added. The DTS was reduced by the inclusion of microspheres. SIGNIFICANCE: PMMA microspheres containing DOX and Ag2SO4 offer a sustained antimicrobial activity for dental applications and promising potential for the biomedical field.

Synthesis and characterization of carboxylic acid conjugated, hydrophobically derivatized, hyperbranched polyglycerols as nanoparticulate drug carriers for cisplatin.

Hyperbranched polyglycerols (HPGs) with hydrophobic cores and derivatized with methoxy poly(ethylene glycol) were synthesized and further functionalized with carboxylate groups to bind and deliver cisplatin. Low and high levels of carboxylate were conjugated to HPGs (HPG-C(8/10)-MePEG(6.5)-COOH(113) and HPG-C(8/10)-MePEG(6.5)-COOH(348)) and their structures were confirmed through NMR and FTIR spectroscopy and potentiometric titration. The hydrodynamic diameter of the HPGs ranged from 5-10 nm and the addition of COOH groups decreased the zeta potential of the polymers. HPG-C(8/10)-MePEG(6.5)-COOH(113) bound up to 10% w/w cisplatin, whereas HPG-C(8/10)-MePEG(6.5)-COOH(348) bound up to 20% w/w drug with 100% efficiency. Drug was released from HPG-C(8/10)-MePEG(6.5)-COOH(113) over 7 days at the same rate, regardless of the pH. Cisplatin release from HPG-C(8/10)-MePEG(6.5)-COOH(348) was significantly slower than HPG-C(8/10)-MePEG(6.5)-COOH(113) at pH 6 and 7.4, but similar at pH 4.5. Release of cisplatin into artificial urine was considerably faster than into buffer. Carboxylated HPGs demonstrated good biocompatibility, and drug-loaded HPGs effectively inhibited proliferation of KU-7-luc bladder cancer cells.

Analysis of the demographics and characteristics of the Australian pharmacist workforce 2013-2018: decreasing supply points to the need for a workforce strategy.

OBJECTIVES: An adequate workforce is necessary for health care delivery. The last official analysis of the Australian pharmacist workforce was in 2014 and the results of recent studies are contradictory. The objective of this work was to determine current demographic details and recent trends of the pharmacy workforce and assess the impact of changes in student numbers and migration policy. METHODS: Longitudinal and descriptive analysis was undertaken of National Health Workforce Datasets and registrant data available from the Australian Health Practitioner Regulation Agency and the Pharmacy Board of Australia from 2013 to 2018. KEY FINDINGS: There was an increase in females and a trend towards hospital practice but no change in the geographic distribution of pharmacists over the period. However, the pharmacist workforce grew more slowly than comparable health professions and while the youngest pharmacist cohort (20-34 years) remains the largest, the next oldest cohort increased at a greater rate. The youngest cohort reported a decrease in intention to remain working in pharmacy. CONCLUSIONS: A fall in student numbers and changes to immigration policy have contributed to a low growth rate and ageing of the pharmacist workforce compared with other professions. Whether these factors along with the intentions of young pharmacists will result in a shortage is dependent on developments in demand for pharmacists and a workforce strategy is required to monitor these developments.

A polymeric paste-drug formulation for local treatment of upper tract urothelial carcinoma.

BACKGROUND: Intravesical instillation of chemo- or immunotherapy is commonly used in bladder cancer. Upper tract urothelial carcinoma (UTUC) shares similar pathological features, but current formulations are not suitable for direct instillation to the upper urinary tract. OBJECTIVE: To evaluate in vivo applicability, characteristics and toxicity of ST-UC, a mucoadhesive polymeric paste formulation of gemcitabine, for upper urinary tract instillation. MATERIAL AND METHODS: Three pigs received 10 ml of ST-UC (100 mg/ml gemcitabine) retrogradely into 1 renal pelvis for pharmacokinetic studies. Four days later, a second injection into the contralateral renal pelvis was followed by serial euthanasia of the pigs and nephroureterectomy after 1, 3, and 6 hours. Adverse effects were monitored. Urine, serum, and tissue gemcitabine concentrations were measured, along with histologic examination of the upper urinary tract. RESULTS: Retrograde instillation of ST-UC was well tolerated with mild, completely receding hydronephrosis. Urine gemcitabine concentrations were highest in the first 3-hour collection interval. Hundred percent of gemcitabine was recovered in the urine within 24 hours. Serum peak concentrations (cmax) of gemcitabine were low at 5.5 µg/ml compared to the 10 to 30 µg/ml levels observed after a single intravenous dose of 1,000 mg/m2 gemcitabine. The formulation was still traceable after one hour and gemcitabine tissue concentrations are supportive of this extended drug exposure. No major histopathological changes were observed. The main limitation of this study is the lack of antitumor activity data. CONCLUSION: This preclinical evaluation of ST-UC demonstrated feasible instillation in the renal pelvis, no significant safety concerns, and sustained release of gemcitabine.

Calcium pyrophosphate dihydrate crystal-induced inhibition of neutrophil apoptosis: involvement of Bcl-2 family members.

INTRODUCTION: The inflammation associated with calcium pyrophosphate dihydrate (CPPD) crystal-induced arthritis arises from the activation of neutrophils with crystals in the synovial joint. Furthermore, constitutive neutrophil apoptosis is inhibited by this interaction with CPPD so that the lifetime of the cells and the duration of the inflammatory response are extended. The objective of this study was to investigate the role of bcl-2 protein family members in the CPPD-induced prosurvival response. METHODS: Apoptosis was measured using DNA fragmentation and Caspase 3 assays. The expression and activation levels of the bcl-2 protein family members A1, Mcl-1, Bcl-xl, Bim, Bad and Bax-alpha were measured using western blot analysis. RESULTS: The prosurvival proteins Mcl-1 and Bcl-xl were both found to be strongly expressed but unaffected by CPPD-induced neutrophil activation over 3 h. The expression of proapoptotic proteins Bim and Bax-alpha was found to decrease over the time course of a 3 h incubation of neutrophils with CPPD crystals (but not the bacterial chemoattractant fMLP). Furthermore, expression of the unphosphorylated (active, proapoptotic) form of Bim was dominant in control cells at 0.5 h, whereas the status of this protein switched to the phosphorylated form following cell activation by both CPPD and fMLP. For CPPD (but not fMLP) this phosphorylation effect reversed over a 3 h incubation. CONCLUSION: Upon stimulation by CPPD crystals, the expression of both Bim and Bax-alpha decreased after 3 h suggesting a reduced proapoptotic effect of these proteins so that the static expression of the prosurvival proteins Bcl-xl and Mcl-1 might allow for a temporary shift in the balance to a prosurvival state of the cells. Because a sudden (but transient) increase in the phosphorylated form of Bim was observed in CPPD-stimulated neutrophils it is possible that this species might act as a signaling intermediate, resulting in the observed downregulation of Bax-alpha.

The inhibition of collagenase induced degradation of collagen by the galloyl-containing polyphenols tannic acid, epigallocatechin gallate and epicatechin gallate.

Collagen based cosmetic fillers require repeat treatments due to collagenase derived degradation of the filler in the intradermal injection site. The objective of this study was to investigate the inhibition of this degradation by the galloyl-containing compounds tannic acid, epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and gallic acid (GA). A gel permeation chromatography assay was developed to quantitate the collagenase induced reductions in collagen molecular weight. The binding of the compounds to collagen was measured using HPLC. The stabilization of collagen was measured using Differential Scanning Calorimetry (DSC). Tannic acid, EGCG and ECG (but not GA) were found to strongly inhibit collagen degradation at concentrations in the low micromolar range. The compounds bound strongly to collagen and stabilized collagen. It is concluded that tannic acid, EGCG and ECG bind to collagen via extensive hydrogen bonding augmented by some hydrophobic interactions and prevent the free access of collagenase to active sites on the collagen chains.

Strategies to promote access to medications during the COVID-19 pandemic.

BACKGROUND: During the COVID-19 pandemic, vulnerable and older people with chronic and complex conditions have self-isolated in their homes, potentially limiting opportunities for consultations to have medications prescribed and dispensed. OBJECTIVE: The aim of this article is to describe initiatives to ensure ongoing access to medications during the COVID-19 pandemic. DISCUSSION: Cooperation between wholesalers and purchase limits in pharmacies have helped to ensure supply of essential medications. Therapeutic substitution by pharmacists is permitted for specific products authorised by the Therapeutic Goods Administration. Prescribers are permitted to issue digital image prescriptions, and implementation of electronic prescribing has been fast-tracked. Expanded continued dispensing arrangements introduced during the bushfire crises have been temporarily extended. Pharmacists are permitted to provide medication management reviews via telehealth. A Home Medicines Service has been introduced to facilitate delivery of medications to people who are vulnerable or elderly. Anticipatory prescribing and medication imprest systems are valuable for access to end-of-life medications within residential aged care.