Anything You Can Do, I Can Do Better: Can Aptamers Replace Antibodies in Clinical Diagnostic Applications?

The mainstay of clinical diagnostics is the use of specialised ligands that can recognise specific biomarkers relating to pathological changes. While protein antibodies have been utilised in these assays for the last 40 years, they have proven to be unreliable due to a number of reasons. The search for the 'perfect' targeting ligand or molecular probe has been slow, though the description of chemical antibodies, also known as aptamers, nearly 30 years ago suggested a replacement reagent. However, uptake has been slow to progress into the clinical environment. In this review, we discuss the issues associated with antibodies and describe some of the applications of aptamers that have relevancy to the clinical diagnostic environment.

Dynamics of active emmetropisation in young chicks--influence of sign and magnitude of imposed defocus.

PURPOSE: Young eyes compensate for the defocus imposed by spectacle lenses by changing their rate of elongation and their choroidal thickness, bringing their refractive status back to the pre-lens condition. We asked whether the initial rate of change either in the ocular components or in refraction is a function of the power of the lenses worn, a result that would be consistent with the existence of a proportional controller mechanism. METHODS: Two separate studies were conducted; both tracked changes in refractive errors and ocular dimensions. Study A: To study the effects of lens power and sign, young chicks were tracked for 4 days after they were fitted with positive (+5, +10 or +15 D) or negative (-5, -10, -15 D) lenses over one eye. In another experiment, biometric changes to plano, +1, +2 and +3 D lenses were tracked over a 24 h treatment period. Study B: Normal emmetropisation was tracked from hatching to 6 days of age and then a defocusing lens, either +6 D or -7 D, was fitted over one eye and additional biometric data collected after 48 h. RESULTS: In study A, animals treated with positive lenses (+5, +10 or +15 D) showed statistical similar initial choroid responses, with a mean thickening 24 µm h(-1) over the first 5 h. Likewise, with the low power positive lenses, a statistically similar magnitude of choroidal thickening was observed across groups (+1 D: 46.0 ± 7.8 µm h(-1); +2 D: 53.5 ± 9.9 µm h(-1); +3 D 53.3 ± 24.1 µm h(-1)) in the first hour of lens wear compared to that of a plano control group. These similar rates of change in choroidal thickness indicate that the signalling response is binary in nature and not influenced by the magnitude of the myopic defocus. Treatments with -5, -10 and -15 D lenses induced statistically similar amounts of choroidal thinning, averaging -70 ± 15 µm after 5 h and -96 ± 45 µm after 24 h. Similar rates in inner axial length changes were also seen with these lens treatments until compensation was reached, once again indicating that the signalling response is not influenced by the magnitude of hyperopic defocus. In study B, after 48 h of +6 D lens treatment, the average refractive error and choroidal changes were found to be larger in magnitude than expected if perfect compensation had taken place, with a + 2.4 D overshoot in refractive compensation. CONCLUSION: Taken together, our results with both weak and higher power positive lenses suggest that eye growth is guided more by the sign than by the magnitude of the defocus, and our results for higher power negative lenses support a similar conclusion. These behaviour patterns and the overshoot seen in Study B are more consistent with the behaviour of a bang-bang controller than a proportional controller.

Demographics and distribution of new entrants to the optometry profession in Australia.

CLINICAL RELEVANCE: An assessment of the total number, demographics and geographic distribution of new entrants to the optometry profession in Australia can assist planning for workforce requirements. BACKGROUND: Over the past decade, the number of registered optometrists in Australia has increased by 30.1 per cent, a rate that is greater than the population growth of the country (12.1 per cent). Concerns have been expressed about the size of the optometry workforce in a context of increasing numbers of graduating optometrists. This paper analyses data obtained from the Australian Health Practitioner Regulation Agency (AHPRA) about new entrants to the profession and their initial practice location during the period 1 July 2010 to 30 June 2018. METHODS: A de-identified dataset was obtained from AHPRA that revealed the following characteristics of new entrants: qualification, gender, year of birth (in five-year bands), registration type, registration endorsement and principal place of practice including its Remoteness Area classification. RESULTS: Data for 1,680 entrants were eligible for analysis; 80 per cent graduated from an Australian university, 12 per cent graduated from the optometry program in New Zealand, and seven per cent were graduates of an overseas university. The remaining two per cent registered via the Trans-Tasman Mutual Recognition Agreement, although the dataset did not include the qualification. The United Kingdom and Republic of Ireland provided the majority of overseas entrants (60 per cent). Most (75 per cent) entrants commenced practice in a major Australian city. Graduates of Australian universities tended to commence practice in the state in which they trained or an adjacent state or territory. Juxtaposed on the data outlined above is the high proportion (42 per cent) of overseas-trained optometrists commencing practice in Western Australia. CONCLUSION: Coincident with the newer optometry programs producing graduates is the increased number of optometrists entering the Australian workforce over the past decade, with the majority commencing practice in major cities. Australia-trained optometrists tend to commence practice in the state where their training was undertaken. New entrants to the optometry profession can be generalised as graduates of an Australian optometry program, female, aged in their early-mid 20s and qualified for therapeutic practice.

Corneal oedema in a unilateral corneal graft patient induced by high Dk mini-scleral contact lens.

INTRODUCTION: Scleral contact lenses are increasingly becoming accepted as the method of choice for visual correction of the irregular cornea. As such, cases have surfaced which demonstrate complications arising from mini-scleral lenses. Identification of these issues and adjusting fitting techniques accordingly is necessary for reducing the risks associated with mini-scleral lens wear. CASE REPORT: A 58 year old Caucasian female was referred for rigid gas permeable contact lens fitting for correction of right irregular astigmatism post penetrating keratoplasty. After four months of successful mini-scleral contact lens wear, the patient experienced a graft rejection episode and treated accordingly, then refit with a new mini-scleral lens. Five months after the lens refit, the patient presented with complaints of hazy vision, and a diagnosis of lens-induced corneal oedema made. DISCUSSION: Increased awareness of the potential complications of mini-scleral lenses is necessary to encourage and enforce mini-scleral lens fitting techniques that meet the requirements of minimum vault but adequate protection of the compromised cornea.

Causes and consequences of a tropical forest gold rush in the Guiana Shield, South America.

Statistical and spatial analyses of both historical time series and remotely sensed data show a link between the spatial distribution and growth of gold production across the Guiana Shield in northeast Amazonia. Results indicate that an exponential rise in production across an expanding area is primarily a delayed response to the 1971-1978 market flotation of international gold prices. The subsequent 10-fold (2-fold) average nominal (real) price increase has provided a compelling economic incentive to mass exploitation of lower-grade gold deposits. The ground-based and remotely sensed distributions of mining activity are strongly attached to these deposits that dominate the region's gold geology. The presence of these gold-bearing formations in conservation and sustainable timber zones has sparked social conflict and environmental degradation across the region. Left unmanaged, more than a quarter-million square-kilometer area of tropical forest zoned for protection and sustainable management could ultimately be compromised by the price-driven boom in gold mining through poorly integrated resource use planning, lack of reclamation effort, and control of illegal operations. Serious public health issues propagated through the unregulated mining environment further erode the financial benefits achieved through gold extraction. This study demonstrates in part how international economic policies successfully stabilizing more conspicuous centers of the global economy can have unintended but profound environmental and social impacts on remote commodity frontiers.

Spatial and temporal patterns of seed attack and germination in a large-seeded neotropical tree species.

It has long been argued that seed dispersal enhances recruitment in tropical trees by allowing offspring to `escape' strong density/distance-dependent attack by insects, pathogens and rodents. Here we examined the effects of canopy openness and parent-offspring distance upon the frequency and timing of Chlorocardium rodiei seed attack and germination within a 15-ha plot of Guyanan tropical rain forest. Seeds were artificially dispersed beneath parent trees, in the understorey away from trees and in gaps. Analysing our data from an 85-week period of regular monitoring, we found that the main spatial gradients, canopy openness and distance to nearest adult conspecific, do not lead to differences in the final number of seeds attacked by infesting scolytid beetles or rodents. The timing of beetle attack, however, varied along the distance gradient and this difference affords seeds at further distances a `window' in which to germinate and produce a seedling before attack. Canopy openness was not a good predictor of rooting success, but distance was strongly associated with root and shoot formation success and the mean time to shoot formation. There was a strong negative effect of distance on the likelihood of a seed being colonised by scolytid beetles prior to removal by rodents and shoot failure was strongly associated with prior infestation. We believe these results bring a key point to bear on the well-established notion of distance-dependent attack on seeds in tropical rainforests, viz. that seed characteristics (size, germination syndrome) and the timing of attack may be more important in explaining patterns of early seedling recruitment than distance. Our studies suggest that advantages accrued through dispersal in species like Chlorocardium will depend heavily on the `race' between seed germination and attack. In the case of Chlorocardium, the `race' can be lost at considerable distances due to its prolonged dormancy and the temporal fluctuations in fruitfall and rainfall which influence attack and germination. The results presented here suggest that the lag between seed attack and germination in tropical trees can regulate the influence of parent-offspring distance on cohort recruitment at this life history stage.

Compensation to positive as well as negative lenses can occur in chicks reared in bright UV lighting.

An earlier report describing a lack of compensation to imposed myopic and hyperopic defocus in chicks reared in UV lighting has led to the belief that the spatial resolving power of the UV cone photoreceptor network in chicks is not capable of decoding optical defocus. However this study used dim light rearing conditions, of less than 10 lx. The purpose of the current study was to determine if emmetropization is possible in young chicks reared under higher luminance, UV lighting conditions. Young, 4 day-old chicks were reared under diurnal near UV (390 nm) illumination set to either 20 or 200 lx while wearing a monocular defocusing lens (+20, +10, -10 or -20 D), for 7 days. Similarly treated control groups were reared under diurnal white lighting (WL) of matching illuminance. The WL and UV LED sources were set to equivalent illuminances, measured in "chick lux", calculated from radiometer readings taken through appropriate narrow band interference filters, and a mathematical model of the spectral sensitivity of the chick visual system. High resolution A-scan ultrasonography was undertaken on days 0 (before lenses were fitted), 2, 4, and 7 to track ocular dimensions and refractive errors were measured by retinoscopy on days 0 and 7. Compensation to negative lenses was unaffected by UV illuminance levels, with near full compensation being achieved under both conditions, as well as under both WL conditions. In contrast, compensation to the positive lenses was markedly impaired in 20 lx UV lighting, with increased instead of decreased axial elongation along with a myopic refractive shift being recorded with the +10D lens. Compensation under both WL conditions was again near normal for the +10D lens. However, with the +20 D lens, myopic shifts in refractive error were observed under both dim UV and WL conditions. The spatial resolving power of the UV cone photoreceptor network in the chick is sufficient to detect optical defocus and guide the emmetropization response, provided illumination is sufficiently high. However, compensation to imposed myopic defocus may be compromised, when either the amount of defocus is very high or illumination low, especially when the wavelength is restricted to the UV range.

Selection of SHV extended-spectrum-beta-lactamase-dependent cefotaxime and ceftazidime resistance in Klebsiella pneumoniae requires a plasmid-borne blaSHV gene.

In Klebsiella pneumoniae, it is common for plasmid-located and chromosome-located bla(SHV) copies to coexist within single cells. The plasmid-borne genes are mainly derived from two separate IS26-mediated mobilizations of bla(SHV). The objective of this study was to test the hypothesis that the presence of a non-extended-spectrum beta-lactamase (non-ESBL) encoding plasmid-borne form of bla(SHV) facilitates the cefotaxime (CTX)-mediated selection of ESBL-expressing mutants, even when there is a chromosomal copy of the same gene. Twenty-one diverse ESBL-negative, bla(TEM)-negative K. pneumoniae clinical isolates were tested for the IS26 insertions characteristic of the two mobilization events. The isolates were then tested for their ability to be selected for ESBL-mediated CTX resistance by serial subculturing with a doubling of the CTX concentration at every subculture. Fourteen isolates possessed neither of the IS26 insertions. None of these became ESBL positive, and all died during the course of the experiment, despite possessing chromosomal bla(SHV) copies. The other isolates all became ESBL positive and grew abundantly up to a CTX concentration of 128 microg/ml. Similar results were obtained with ceftazidime. ESBL expression was associated with the appearance of the expected G-->A mutation at position 1 of codon 238 and also with bla(SHV) copy number amplification. It was concluded that plasmid-borne bla(SHV) greatly facilitates the selection of ESBL expression, even when the same gene is on the chromosome, and that gene dosage effects are likely to contribute to this phenomenon.

bla(SHV) Genes in Klebsiella pneumoniae: different allele distributions are associated with different promoters within individual isolates.

Extended-spectrum beta-lactamases (ESBLs) emerge by point mutation from non-extended-spectrum precursors. The aims of this study were to reveal the basis for variations in resistance levels found in a collection of 21 Klebsiella pneumoniae clinical isolates from Brisbane, Australia. Previous studies have shown that 20 of these isolates possess bla(SHV-11), bla(SHV-2a), and/or bla(SHV-12), and there is an association between the copy numbers of the ESBL-encoding genes and resistance levels. In this study, a real-time PCR method for interrogating the polymorphic sites at codons 238 and 240 was developed, and this confirmed the relationship between mutant gene copy numbers and resistance levels. The bla(SHV) promoter region was cloned from one of the ESBL-expressing isolates, and this showed that bla(SHV) genes exist downstream of two different promoters within this single isolate. These promoters have both been reported previously, and they differ by virtue of the presence or absence of an IS26 insertion. The bla(SHV) copy numbers in cis with the different promoters were measured, and the copy number of the IS26 promoter was correlated with resistance levels. Cloning and analysis of PCR products showed that different bla(SHV) variants existed in cis with individual promoters in individual isolates but that mutant genes were more abundant downstream of the IS26 promoter. There were no ESBL-positive isolates without this promoter. It was concluded that bla(SHV) in cis with the IS26 promoter is located on an amplifiable replicon, and the presence of the IS26 insertion may facilitate the acquisition of an ESBL-positive phenotype.