Parents' perceptions of microneedle-mediated monitoring as an alternative to blood sampling in the monitoring of their infants

OBJECTIVES: Microneedle (MN) arrays could offer a pain-free, minimally invasive approach to monitoring. This is envisaged to be particularly beneficial for younger patients, but parents' views to date are unknown. The aim of this study was to explore parental perceptions of MN-mediated ISF monitoring, as an alternative to the use of conventional blood sampling, and to understand the important factors for technique approval.

METHODS: Semi-structured interviews were conducted with parents with recent experience of a premature birth. Recruitment was through the Northern Ireland premature infant charity, Tinylife. Interviews progressed until data saturation was reached and thematic analysis employed.

KEY FINDINGS: The study included 16 parents. Parental support for MN-mediated monitoring was evident, alongside the unpopularity of traditional blood sampling in neonates. Factors facilitating MN approval included the opportunity for pain reduction, the simplicity of the procedure, the potential for increased parental involvement and the more favourable appearance, owing to the minute size of MNs and similarities with a sticking plaster. Confirmation of correct application, a pain-free patch removal and endorsement from trusted healthcare professionals were important.

CONCLUSION: These findings will inform researchers in the field of MN development and enlighten practitioners regarding parental distress resulting from conventional blood sampling. Further work is necessary to understand MN acceptability among practitioners. This work should assist in the development of an acceptable MN device and facilitate the reduction of parental distress.

Design and physicochemical characterisation of novel dissolving polymeric microneedle arrays for transdermal delivery of high dose, low molecular weight drugs

We describe formulation and evaluation of novel dissolving polymeric microneedle (MN) arrays for the facilitated delivery of low molecular weight, high dose drugs. Ibuprofen sodium was used as the model here and was successfully formulated at approximately 50% w/w in the dry state using the copolymer poly(methylvinylether/maleic acid). These MNs were robust and effectively penetrated skin in vitro, dissolving rapidly to deliver the incorporated drug. The delivery of 1.5mg ibuprofen sodium, the theoretical mass of ibuprofen sodium contained within the dry MN alone, was vastly exceeded, indicating extensive delivery of the drug loaded into the baseplates. Indeed in in vitro transdermal delivery studies, approximately 33mg (90%) of the drug initially loaded into the arrays was delivered over 24h. Iontophoresis produced no meaningful increase in delivery. Biocompatibility studies and in vivo rat skin tolerance experiments raised no concerns. The blood plasma ibuprofen sodium concentrations achieved in rats (263μgml(-1) at the 24h time point) were approximately 20 times greater than the human therapeutic plasma level. By simplistic extrapolation of average weights from rats to humans, a MN patch design of no greater than 10cm(2) could cautiously be estimated to deliver therapeutically-relevant concentrations of ibuprofen sodium in humans. This work, therefore, represents a significant progression in exploitation of MN for successful transdermal delivery of a much wider range of drugs.

High-affinity NO3 --H+ cotransport in the fungus Neurospora:Induction and control by pH and membrane voltate

High-affinity nitrate transport was examined in intact hyphae of Neurospora crassa using electrophysiological recordings to characterize the response of the plasma membrane to NO₃ ^- challenge and to quantify transport activity. The NO₃ ^--associated membrane current was determined using a three electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in hyphae transferred to NO₃ ^--free, N-limited medium for 15 hr, and in hyphae grown in the absence of a nitrogen source after a single 2-min exposure to 100 μM NO₃ ^-. In the latter, induction showed a latency of 40-80 min and rose in scalar fashion with full transport activity mensurable approx. 100 min after first exposure to NO₃ ^-; it was marked by the appearance of a pronounced sensitivity of membrane voltage to extracellular NO₃ ^- additions which, after induction, resulted in reversible membrane depolarizations of (+)54-85 mV in the presence of 50 μM NO₃ ^-; and it was suppressed when NH₄ ⁺, was present during the first, inductive exposure to NO₃ ^-. Voltage clamp measurements carried out immediately before and following NO₃ ^- additions showed that the NO₃ ^--evoked depolarizations were the consequence of an inward-directed current that appeared in parallel with the depolarizations across the entire range of accessible voltages -400 to +100 mV). Measurements of NO₃ ^- uptake using NO₃ ^--selective macroelectrodes indicated a charge stoichiometry for NO₃ ^- transport of 1(+):1(NO₃ ^-) with common K(m) and J(max) values around 25 μM and 75 pmol NO₃ ^- cm^-2sec^-1, respectively, and combined measurements of pH(o) and [NO₃ ^-](o) showed a net uptake of approx. 1 H⁺ with each NO₃ ^- anion. Analysis of the NO₃ ^- current demonstrated a pronounced voltage sensitivity within the normal physiological range between -300 and -100 mV as well as interactions between the kinetic parameters of membrane voltage, pH(o) and [NO₃ ^-](o). Increasing the bathing pH from 5.5 to 8.0 reduced the current and the associated membrane depolarizations 2- to 4-fold. At a constant pH(o) of 6.1, driving the membrane voltage from -350 to -150 mV resulted in an approx. 3-fold reduction in the maximum current and a 5-fold rise in the apparent affinity for NO₃ ^-. By contrast, the same depolarization effected an approx. 20% fall in the K(m) for transport as a function in [H⁺](o). These, and additional results are consistent with a charge-coupling stoichiometry of 2(H⁺) per NO anion transported across the membrane, and implicate a carrier cycle in which NO binding is kinetically adjacent to the rate-limiting step of membrane charge transit. The data concur with previous studies demonstrating a pronounced voltage-dependence to high-affinity NO₃ ^- transport system in Arabidopsis, and underline the importance of voltage as a kinetic factor controlling NO₃ ^- transport; finally, they distinguish metabolite repression of NO₃ ^- transport induction from its sensitivity to metabolic blockade and competition with the uptake of other substrates that draw on membrane voltage as a kinetic substrate.

Cable correction of membrane currents recorded from root hairs of Arabidopsis thaliana L.

Membrane currents were recorded under voltage clamp from root hairs of Arabidopsis thaliana L. using the two-electrode method. Concurrent measurements of membrane voltage distal to the point of current injection were also carried out to assess the extent of current dissipation along the root hair axis. Estimates of the characteristic cable length, λ, showed this parameter to be a function both of membrane voltage and of substrate concentration for transport. The mean value for λ at 0 mV was 103 ± 20 μm (n=17), but ranged by as much as 6-fold in any one cell for membrane voltages from -300 to +40 mV and was affected by 0.25 to 3-fold at any one voltage on raising [K⁺]₀ from 0.1 to 10 mol m^-3. Current dissipation along the length of the cells lead to serious distortions of the current-voltage [I-V) characteristic, including consistent underestimates of membrane current as well as a general linearization of the I-V curve and a masking of conductance changes in the presence of transported substrates. In some experiments, microelectrodes were also placed in neighbouring epidermal cells to record the extent of intercellular coupling. Even with current-passing microelectrodes placed at the base of root hairs, coupling was ≤5% (voltage deflection of the epidermal cell ≤5% that recorded at the site of current injection), indicating an appreciable resistance to current passage between cells. These results demonstrate the feasibility of using root hairs as a 'single-cell model' in electrophysiological analyses of transport across the higher-plant plasma membrane; they also confirmed the need to correct for the cable properties of these cells on a cell-by-cell basis. © 1994 Oxford University Press.

Role of early visual cortex in trans-saccadic memory of object features

Early visual cortex (EVC) participates in visual feature memory and the updating of remembered locations across saccades, but its role in the trans-saccadic integration of object features is unknown. We hypothesized that if EVC is involved in updating object features relative to gaze, feature memory should be disrupted when saccades remap an object representation into a simultaneously perturbed EVC site. To test this, we applied transcranial magnetic stimulation (TMS) over functional magnetic resonance imaging-localized EVC clusters corresponding to the bottom left/right visual quadrants (VQs). During experiments, these VQs were probed psychophysically by briefly presenting a central object (Gabor patch) while subjects fixated gaze to the right or left (and above). After a short memory interval, participants were required to detect the relative change in orientation of a re-presented test object at the same spatial location. Participants either sustained fixation during the memory interval (fixation task) or made a horizontal saccade that either maintained or reversed the VQ of the object (saccade task). Three TMS pulses (coinciding with the pre-, peri-, and postsaccade intervals) were applied to the left or right EVC. This had no effect when (a) fixation was maintained, (b) saccades kept the object in the same VQ, or (c) the EVC quadrant corresponding to the first object was stimulated. However, as predicted, TMS reduced performance when saccades (especially larger saccades) crossed the remembered object location and brought it into the VQ corresponding to the TMS site. This suppression effect was statistically significant for leftward saccades and followed a weaker trend for rightward saccades. These causal results are consistent with the idea that EVC is involved in the gaze-centered updating of object features for trans-saccadic memory and perception.

Is the Schwabe organ a retained larval eye? Anatomical and behavioural studies of a novel sense organ in adult leptochiton asellus (Mollusca, Polyplacophora) indicate links to larval photoreceptors

The discovery of a sensory organ, the Schwabe organ, was recently reported as a unifying feature of chitons in the order Lepidopleurida. It is a patch of pigmented tissue located on the roof of the pallial cavity, beneath the velum on either side of the mouth. The epithelium is densely innervated and contains two types of potential sensory cells. As the function of the Schwabe organ remains unknown, we have taken a cross-disciplinary approach, using anatomical, histological and behavioural techniques to understand it. In general, the pigmentation that characterises this sensory structure gradually fades after death; however, one particular concentrated pigment dot persists. This dot is positionally homologous to the larval eye in chiton trochophores, found in the same neuroanatomical location, and furthermore the metamorphic migration of the larval eye is ventral in species known to possess Schwabe organs. Here we report the presence of a discrete subsurface epithelial structure in the region of the Schwabe organ in Leptochiton asellus that histologically resembles the chiton larval eye. Behavioural experiments demonstrate that Leptochiton asellus with intact Schwabe organs actively avoid an upwelling light source, while Leptochiton asellus with surgically ablated Schwabe organs and a control species lacking the organ (members of the other extant order, Chitonida) do not (Kruskal-Wallis, H = 24.82, df = 3, p < 0.0001). We propose that the Schwabe organ represents the adult expression of the chiton larval eye, being retained and elaborated in adult lepidopleurans.

A Largest Matching Area Approach to Image Denoising

Given the success of patch-based approaches to image denoising,this paper addresses the ill-posed problem of patch size selection.Large patch sizes improve noise robustness in the presence of good matches, but can also lead to artefacts in textured regions due to the rare patch effect; smaller patch sizes reconstruct details more accurately but risk over-fitting to the noise in uniform regions. We propose to jointly optimize each matching patch’s identity and size for gray scale image denoising, and present several implementations.The new approach effectively selects the largest matching areas, subject to the constraints of the available data and noise level, to improve noise robustness. Experiments on standard test images demonstrate our approach’s ability to improve on fixed-size reconstruction, particularly at high noise levels, on smoother image regions.

Microneedle-mediated delivery of donepezil: Potential for improved treatment options in Alzheimer's disease

Transdermal drug delivery is an attractive route of drug administration, however there are relatively few marketed transdermal products. To increase delivery across the skin, strategies to enhance skin permeability are widely investigated, with microneedles demonstrating particular promise. Hydrogel-forming microneedles are inserted into the skin, and following dissolution of a drug loaded reservoir and movement of the drug through the created channels, the microneedle array is removed intact, and can then be readily and safely discarded. This study presents the formulation and evaluation of an integrated microneedle patch containing the Alzheimer's drug, donepezil hydrochloride. The integrated patch consisted of hydrogel-forming microneedles in combination with a donepezil hydrochloride containing film. Formulation and characterisation of plasticised films, prepared from poly(vinylpyrrolidone) or poly (methyl vinyl ether co-maleic anhydride/acid) (Gantrez(®)) polymers, is presented. Furthermore, in vitro permeation of donepezil hydrochloride across neonatal porcine skin from the patches was investigated, with 854.71 μg ± 122.71 μg donepezil hydrochloride delivered after 24 h, using the optimum patch formulation. Following administration of the patch to an animal model, plasma concentrations of 51.8 ± 17.6 ng/mL were obtained, demonstrating the success of this delivery platform for donepezil hydrochloride.

Transcending epithelial and intracellular biological barriers; a prototype DNA delivery device

Microneedle technology provides the opportunity for the delivery of DNA therapeutics by a non-invasive, patient acceptable route. To deliver DNA successfully requires consideration of both extra and intracellular biological barriers. In this study we present a novel two tier platform; i) a peptide delivery system, termed RALA, that is able to wrap the DNA into nanoparticles, protect the DNA from degradation, enter cells, disrupt endosomes and deliver the DNA to the nucleus of cells ii) a microneedle (MN) patch that will house the nanoparticles within the polymer matrix, breach the skin's stratum corneum barrier and dissolve upon contact with skin interstitial fluid thus releasing the nanoparticles into the skin. Our data demonstrates that the RALA is essential for preventing DNA degradation within the poly(vinylpyrrolidone) (PVP) polymer matrix. In fact the RALA/DNA nanoparticles (NPs) retained functionality when in the MN arrays after 28days and over a range of temperatures. Furthermore the physical strength and structure of the MNs was not compromised when loaded with the NPs. Finally we demonstrated the effectiveness of our MN-NP platform in vitro and in vivo, with systemic gene expression in highly vascularised regions. Taken together this 'smart-system' technology could be applied to a wide range of genetic therapies.

Ecology and conservation of felids in the Brazilian pantanal

Habitat conversion and environmental degradation have reached alarming levels in the Pantanal, endangering all its biodiversity. This scenario is complicated by the fact that the biome relies on only a few protected areas, which combined do not exceed 10% of the territory. Felids, as predators, play a vital role in the maintenance of this ecosystem, but require large areas, have low population densities and, typically, are very sensitive to environmental disturbances. Amolar Mountain Ridge is considered an area of extreme importance and high priority for conservation within the biome. There are four species of felids in this region: the jaguar (Panthera onca), the puma (Puma concolor), the ocelot (Leopardus pardalis), and jaguarundi (Puma yagouaroundi). However, little is known about the ecology of these species in this region or the magnitude of interaction between the communities living around the protected areas and the animals. The goal of this study was to increase our knowledge about these felids and understand how people interact with them in order to contribute to their conservation in the network of parks within Amolar. Camera trapping surveys were carried out in two areas of the network, covering approximately 83,000 hectares, in order to identify the species of mammals occurring in the region, those that may be potential prey for the felids, and to obtain basic ecological data about both felids and prey. In addition, we conducted surveys in three riverside schools in order to assess the knowledge, perceptions and attitudes of schoolchildren regarding the four focal felids, and surveys among the adult population to assess their perceptions and attitudes towards the jaguar. We recorded a total of 33 species of mammals from both study areas. The large cats were cathemeral, reflecting the temporal activity of larger prey, whereas the ocelot was nocturnal, mirroring the activity of smaller prey. Jaguar occupancy was influenced by prey abundance, while puma occupancy was influenced by patch density in drier dense forest. Jaguars and pumas may be competitors over temporal and spatial scales, while no resource overlap was found for ocelots. Overall, both adults and children tended to have negative perceptions about the cats, which were related to the fear of being attacked. To increase awareness about the species and to maximize the effectiveness of protective measures in the network of reserves, it is recommended to develop and implement an Environmental Educational Program in the medium- to long-term in order to minimize the fear of these felids and to counsel locals on the role of felids in the maintenance of the Pantanal’s biodiversity.

Shape control of laminated panels using piezoelectric actuators

This paper presents a finite element formulation based on the classical laminated plate theory for laminated structures with integrated piezoelectric layers or patches, acting as actuators.The finite element model is a single layer trinaguular nonconforming plate/shell element with 18 degrees of fredom for the generalized displacements, and one electrical potential degree of freedom for each piezoelectric element elemenet layer or patch. An optimization of the patches position is perfomed to maximize the piezoelectric actuators efficiency as well as,the electric potential distribution is serach to reach the specified strusctura transverse displacement distribution is search to reach the specified structures trsnsverse displacement distribution (shape control). A gradient based algorithm is used for this purpose.Results are presented and discussed.

Non-linear analysis of composite structures with integrated piezoelectric sensors and actuators

This paper deals with the geometrically non linear analysis of thin plate/shell laminated structures with embedded integrated piezoelectric actuors or sensors layers and/or patches.The model is based on the Kirchhoff classical laminated theory and can be applied to plate and shell adaptive structures with arbitrary shape, general mechanical and electrical loadings. the finite element model is a nonconforming single layer triangular plate/shell element with 18 degrees of fredom for the generalized displacements and one eçlectrical potential degree of freedom for each piezoelectric layer or patch. An updated Lagrangian formulation associated to Newton-Raphson technique is used to solve incrementally and iteratively the equilibrium equation.The model is applied in the solution of four illustrative cases, and the results are compared and discussedwith alternative solutions when available.

Design of laminated structures using piezoelectric materials

Composite structures incorporating piezoelectric sensors and actuators are increasingly becoming important due to the offer of potential benefits in a wide range of engineering applications such as vibration and noise supression, shape control and precisition positioning. This paper presents a finit element formulation based on classical laminated plate theory for laminated structures with integrated piezoelectric layers or patches, acting as actuators. The finite element model is a single layer triangular nonconforming plate/shell element with 18 degrees of freedom for the generalized displacements, and one electrical potential degree of freedom for each piezsoelectric elementlayer or patch, witch are surface bonded on the laminate. An optimization of the patches position is performed to maximize the piezoelectric actuators efficiency as well as, the electric potential distribuition is search to reach the specified structure transverse displacement distribuition (shape control). A gradient based algorithm is used for this purpose. The model is applied in the optimization of illustrative laminated plate cases, and the results are presented and discussed.

Caracterização dos mecanismos de transporte epitelial no intestino da dourada, Sparus auratus

Dissertação de mest., Aquacultura e Pescas, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2010

The function of drosophila integrin adhesion complexes in pupal hemocyte migration in vivo

Tese de doutoramento, Ciências Biomédicas (Biologia Celular e Molecular), Universidade de Lisboa, Faculdade de Medicina, 2014