Ga Jacobson

Toenail onychomycosis: an important global disease burden

Onychomycosis is a fungal infection of the nail plate or nail bed. It does not usually cure itself and it can trigger more infectious lesions in other parts of the body. The reported prevalence of onychomycosis is increasing in Western countries, presumably due to lifestyle changes and the ageing of the population. Approximately 10% of the general population, 20% of the population aged >60 years, up to 50% of people aged >70 years and up to one‐third of diabetic individuals have onychomycosis. Care should be taken for the accurate diagnosis and timely treatment of toenail onychomycosis to prevent complications. Current treatment options have relatively limited therapeutic success, particularly long‐term. Oral medications are associated with high recurrence rates and treatment failure, and are not suitable for many cases due to potential adverse effects. Topical medications are recommended only for mild to moderate cases. The cost of therapies may also be prohibitive in some cases. In the light of these issues, more research is warranted for the investigation and development of more effective and economical options for the treatment and prophylaxis of toenail onychomycosis. In patient populations such as diabetic individuals, where onychomycosis can provoke lower extremity complications, professional podiatry care of toenails and feet should be encouraged.

Bicycle injuries: road trauma is not the only concern.

Data on bicycle injury presentations at a public hospital emergency department for 1991–95 inclusive were analysed to determine the nature and extent of bicycle injuries in the community. There were 599 bicycle injury presentations during the study period, representing more than 2.0% of all injury‐related presentations to the emergency department. The main outcome measures were severity and type of injury. Distribution by gender, age, helmet use (1991 and 1992 only), location and mechanism was examined. Rider‐only injuries (falls or collisions with stationary objects) accounted for 79.3% of all presentations with only 5.2% due to collisions on a public road or footpath with other moving traffic. Of all injuries where a location was determined, 61.6% occurred in off‐road locations. There was little difference in the overall proportions of hospital admissions from injuries on‐road (12.4%) and off‐road (9.0%). Cyclists injured in on‐road collisions with traffic had a higher proportion of hospital admissions (40.0%) than those injured on‐road by other mechanisms (7.6%). Children under 10 years of age who had been riding without a helmet suffered a much higher proportion of injuries to the head (53.2% of all injuries) than older cyclists riding without a helmet (19.4%). The majority (83.1%) of head injuries in children under 10 years of age occurred off‐road and helmet use was lowest in this group (28.6%). Given previous evidence that helmet use can prevent head injuries, strategies to increase helmet use among cyclists, particularly young children, while riding both on and off‐road, should be given a high priority. Rider‐only injuries are also an important public health issue.

High-performance liquid chromatographic assay for the simultaneous determination of ipratropium bromide, fenoterol, salbutamol and terbutaline in nebulizer solution

A reversed-phase ion-pair high-performance liquid chromatography assay was developed for the simultaneous determination of ipratropium bromide, fenoterol hydrobromide, salbutamol sulphate and terbutaline sulphate in nebulizer solution. Chromatographic separation was achieved with a Nova-Pak C18 4 microns 10 cm x 8 mm i.d. Radial-pak cartridge inside a Waters RCM 8 x 10 compression module using ternary gradient analysis. Detection was performed using UV detection at 220 nm. The standard curves were linear over the following ranges: ipratropium bromide 20.8-250.0 micrograms ml-1, fenoterol hydrobromide 27.8-500.0 micrograms ml-1, salbutamol sulphate 34.7-2500.0 micrograms ml-1 and terbutaline sulphate 69.5-2500 micrograms ml-1. Inter-day and intra-day relative standard deviations for each compound ranged from 4.5-5.2% and 3.5-3.9%, respectively. The assay procedure was developed to allow the accurate determination of constituents in various combinations of nebulizer solution, as well as for stability indicating purposes. This provides a convenient means of testing long-term compatibility and stability following the post-manufacture mixing of commonly used nebulized preparations.

Zebrafish - on the move towards ophthalmological research

Millions of people are affected by visual impairment and blindness globally, and the prevalence of vision loss is likely to increase as we are living longer. However, many ocular diseases remain poorly controlled due to lack of proper understanding of the pathogenesis and the corresponding lack of effective therapies. Consequently, there is a major need for animal models that closely mirror the human eye pathology and at the same time allow higher-throughput drug screening approaches. In this context, zebrafish as an animal model organism not only address these needs but can in many respects reflect the human situation better than the current rodent models. Over the past decade, zebrafish have become an established model to study a variety of human diseases and are more recently becoming a valuable tool for the study of human ophthalmological disorders. Many human ocular diseases such as cataract, glaucoma, diabetic retinopathy, and age-related macular degeneration have already been modelled in zebrafish. In addition, zebrafish have become an attractive model for pre-clinical drug toxicity testing and are now increasingly used by scientists worldwide for the discovery of novel treatment approaches. This review presents the advantages and uses of zebrafish for ophthalmological research.

Proteoglycans in the central nervous system: role in development, neural repair, and Alzheimer's disease.

Proteoglycans (PGs) are major components of the cell surface and extracellular matrix and play critical roles in development and maintenance of the central nervous system (CNS). PGs are a family of proteins, all of which contain a core protein to which glycosaminoglycan side chains are covalently attached. PGs possess diverse physiological roles, particularly in neural development, and are also implicated in the pathogenesis of neurodegenerative diseases such as Alzheimers disease (AD). The main functions of PGs in the CNS are reviewed as are the roles of PGs in brain injury and in the development or treatment of AD.

Effective reversed-phase ion pair high-performance liquid chromatography method for the separation and characterization of intact low-molecular-weight heparins

A simple, selective, and efficient reversed-phase ion pair high-performance liquid chromatography (RPIP-HPLC) method was developed for the separation of various commercially available intact low-molecular-weight heparins (LMWHs). The developed method uses a C(18) column (150 x 4.6 mm) with diode array detection at 230 nm, flow rate at 1.0 ml/min, and a mobile phase containing acetonitrile/water (32:68%), tetrabutylammonium hydroxide (15 mM), and ammonium acetate (50 mM) at pH 7.0. The performance of this method was assessed in terms of selectivity, linearity, intra- and interday precision, and accuracy. The novel application of RPIP-HPLC with evaporative light scattering detection (ELSD) for the analysis of intact LMWHs was demonstrated. Intact LMWHs were analyzed with superior resolution and peak shape. Different chromatographic profiles were obtained for different LMWHs showing significant structural diversity. This method clearly showed chemical changes that occurred to LMWH under the stress condition. This method can be applied for the separation, identification, characterization, and pharmaceutical stability analysis of various LMWHs.

LC-MS method for the determination of albuterol enantiomers in human plasma using manual solid-phase extraction and a non-deuterated internal standard

A sensitive enantioselective liquid chromatography-mass spectrometry (LC-MS) assay using a manual solid-phase extraction (SPE) procedure, a non-deuterated internal standard and an ion trap LC-MS was developed to measure (R)- and (S)-albuterol in plasma. Sample extraction from plasma was achieved by a manual SPE extraction procedure with methoxyphenamine added as the internal standard. Chiral separation was achieved using a teicoplanin-based stationary phase and a mobile phase consisting of methanol, acetic acid and 28% (w/v) ammonia (1000:5:1, v/v/v). Samples were analyzed by selected reaction monitoring of product ions from the protonated molecular ions. The detection limit of the assay was 0.1 ng/ml with a conservative lower limit of quantification of 0.25 ng/ml for each enantiomer. Recovery of albuterol enantiomers from plasma spiked at 10 ng/ml of racemate was determined to be 89+/-5.8% (mean+/-S.D.). Reproducibility at 10 ng/ml of racemate assessed by the coefficient of variation was found to be 6.5% (n=5). Instrument precision (measured as coefficient of variation) was 1.4% (n=5). The correlation coefficient r(2) determined from the calibration curve over the range 0.5-50.0 ng/ml racemate in plasma was 0.998. This assay allows adequate sensitivity, recovery and reproducibility for the application to studies of inhaled albuterol.

Effects of heparin and enoxaparin on APP processing and Aβ production in primary cortical neurons from Tg2576 mice

Background Alzheimers disease (AD) is caused by accumulation of Aβ, which is produced through sequential cleavage of β-amyloid precursor protein (APP) by the β-site APP cleaving enzyme (BACE1) and γ-secretase. Enoxaparin, a low molecular weight form of the glycosaminoglycan (GAG) heparin, has been reported to lower Aβ plaque deposition and improve cognitive function in AD transgenic mice. Methodology/Principal Findings We examined whether heparin and enoxaparin influence APP processing and inhibit Aβ production in primary cortical cell cultures. Heparin and enoxaparin were incubated with primary cortical cells derived from Tg2576 mice, and the level of APP and proteolytic products of APP (sAPPα, C99, C83 and Aβ) was measured by western blotting. Treatment of the cells with heparin or enoxaparin had no significant effect on the level of total APP. However, both GAGs decreased the level of C99 and C83, and inhibited sAPPα and Aβ secretion. Heparin also decreased the level of β-secretase (BACE1) and α-secretase (ADAM10). In contrast, heparin had no effect on the level of ADAM17. Conclusions/Significance The data indicate that heparin and enoxaparin decrease APP processing via both α- and β-secretase pathways. The possibility that GAGs may be beneficial for the treatment of AD needs further study.

Elevated plasma glutathione peroxidase concentration in acute severe asthma: Comparison with plasma glutathione peroxidase activity, selenium and malondialdehyde

Objective. To investigate plasma glutathione peroxidase concentration, glutathione peroxidase activity, plasma selenium and oxidative stress in acute severe asthma. Material and methods. The study was case‐control in design, with cases presenting to the emergency department with acute severe asthma and controls randomly selected from a larger cross‐sectional study. Plasma malondialdehyde (MDA) was used as a measure of oxidative stress and plasma selenium was measured using ICP‐MS. Glutathione peroxidase (GPx) activity was analysed using a colorimetric GPx assay and plasma GPx level was measured by enzyme‐linked immunosorbent assay (ELISA). Results. Fifteen cases [mean (range) predicted peak expiratory flow rate (PEFR) of 43% (20–69)] and 15 matched controls were recruited. MDA levels (mean±SD) were higher in acute asthma subjects (1.30±0.56 µmol/L) than in controls (0.86±0.53 µmol/L; p<0.05). There were no differences between cases and controls for selenium (99±34 µg/L versus 109±17 µg/L) or for GPx activity (39±25 nmol min−1 mL−1 versus 38±24 nmol min−1 mL−1), however, GPx plasma levels measured by ELISA were higher in cases than controls (22.5±10.8 mg/L versus 13.8±7.3 mg/L; p<0.05). Conclusions. Patients with acute severe asthma demonstrated increased MDA levels but no differences in plasma selenium levels or GPx activity. GPx levels measured by ELISA were elevated in severe asthma. These results are consistent with an adaptive up‐regulation of GPx to protect against oxidative stress.

Investigation of the Effect of Heating on the Chemistry and Antifactor Xa Activity of Enoxaparin

The objective of this study was to investigate the effects of heating on the chemistry, physical properties and antifactor Xa activity of enoxaparin. Samples of enoxaparin heated at 70 degrees C lost 27% of their initial AFXa activity after 8 h, then activity increased to 94% of the initial activity over the next 4 h. Activity then decreased to 84% of control after 48 h and further to 80% of control over 22 days. The initial activity loss correlated with desulfation as demonstrated by sulfate and amine analysis. Fragmentation of oligosaccharides occurred, as demonstrated by reducing capacity and capillary electrophoresis analysis. Individual enoxaparin fractions obtained by high performance size exclusion chromatography were analysed. Early eluting fractions, containing aggregated oligosaccharides, increased in concentration following heating. Up to 65% of sulfate was lost from some fractions, containing hexa- and octa-saccharides, after 8 h, corresponding with decreased activity. Low mass oligosaccharide fractions increased in concentration and had increased activity between 8 and 12 h. Reversed-phase ion-interaction HPLC analysis supported these findings. Deca-, dodeca- and tetradeca-saccharides were resistant to thermal degradation. Desulfation, fragmentation and aggregation occur during the heating of enoxaparin and result in the initial rapid loss, recovery and subsequent gradual loss of activity.