Methylene Blue-Loaded Dissolving Microneedles: Potential Use in Photodynamic Antimicrobial Chemotherapy of Infected Wounds

Photodynamic therapy involves delivery of a photosensitising drug that is activated by light of a specific wavelength, resulting in generation of highly reactive radicals. This activated species can cause destruction of targeted cells. Application of this process for treatment of microbial infections has been termed "photodynamic antimicrobial chemotherapy" (PACT). In the treatment of chronic wounds, the delivery of photosensitising agents is often impeded by the presence of a thick hyperkeratotic/necrotic tissue layer, reducing their therapeutic efficacy. Microneedles (MNs) are an emerging drug delivery technology that have been demonstrated to successfully penetrate the outer layers of the skin, whilst minimising damage to skin barrier function. Delivering photosensitising drugs using this platform has been demonstrated to have several advantages over conventional photodynamic therapy, such as, painless application, reduced erythema, enhanced cosmetic results and improved intradermal delivery. The aim of this study was to physically characterise dissolving MNs loaded with the photosensitising agent, methylene blue and assess their photodynamic antimicrobial activity. Dissolving MNs were fabricated from aqueous blends of Gantrez(®) AN-139 co-polymer containing varying loadings of methylene blue. A height reduction of 29.8% was observed for MNs prepared from blends containing 0.5% w/w methylene blue following application of a total force of 70.56 N/array. A previously validated insertion test was used to assess the effect of drug loading on MN insertion into a wound model. Staphylococcus aureus, Escherichia coli and Candida albicans biofilms were incubated with various methylene blue concentrations within the range delivered by MNs in vitro (0.1-2.5 mg/mL) and either irradiated at 635 nm using a Paterson Lamp or subjected to a dark period. Microbial susceptibility to PACT was determined by assessing the total viable count. Kill rates of >96%, were achieved for S. aureus and >99% for E. coli and C. albicans with the combination of PACT and methylene blue concentrations between 0.1 and 2.5 mg/mL. A reduction in the colony count was also observed when incorporating the photosensitiser without irradiation, this reduction was more notable in S. aureus and E. coli strains than in C. albicans.

SPLI is a Chemoattractant for Oral and Skin Fibroblasts

Objectives: Unlike adult dermal wounds, the oral mucosa demonstrates preferential healing characterized by rapid remodeling and re-epithelialisation, with minimal scar formation. Secretory leukocyte protease inhibitor (SLPI) is an epithelial-derived factor with potential for promoting scarless repair. The aims of this study were to: (i) investigate the directed migratory (chemotaxis) response of oral and skin fibroblasts to various concentrations of SLPI; and (ii) compare migratory speed of the two cell types. Methods: Paired oral and skin fibroblasts were seeded at 2x104 cells in six well plates containing glass coverslips, and cultured in DMEM supplemented with 10% FCS for 48hours. Following a period of serum starvation (18hours in DMEM plus 0.5% BSA), coverslips were incorporated within a Dunn chemotaxis chamber containing DMEM with 0.5% BSA +/- SLPI gradients at 0.5, 1 or 2µM concentrations. Using microscopy, the migratory behaviour of cells was digitally captured every 10mins for 18hours, traced with JCell tracking software and resulting co-ordinates statistically analysed using Mathmatica software. Results: At all concentrations SLPI was a significant chemoattractant (p<0.01) for both cell types. However, skin fibroblasts migrated significantly faster than oral cells at each SLPI concentration, with greatest effect observed at the highest dose (skin: 32.0±0.47µm/hr, oral: 13.6±0.23µm/hr). Conclusion: SLPI is a chemoattractant for both oral and skin fibroblasts, and may play an important role in fibroblast recruitment during wound healing. This work was funded by the R&D Office, N.Ireland.

Myopia, spectacle wear, and risk of bicycle accidents among rural Chinese secondary school students: the Xichang Pediatric Refractive Error Study report no. 7.

OBJECTIVE: To study the effect of myopia and spectacle wear on bicycle-related injuries in rural Chinese students. Myopia is common among Chinese students but few studies have examined its effect on daily activities. METHODS: Data on visual acuity, refractive error, current spectacle wear, and history of bicycle use and accidents during the past 3 years were sought from 1891 students undergoing eye examinations in rural Guangdong province. RESULTS: Refractive and accident data were available for 1539 participants (81.3%), among whom the mean age was 14.6 years, 52.5% were girls, 26.8% wore glasses, and 12.9% had myopia of less than -4 diopters in both eyes. More than 90% relied on bicycles to get to school daily. A total of 2931 accidents were reported by 423 participants, with 68 requiring medical attention. Male sex (odds ratio, 1.55; P < .001) and spectacle wear (odds ratio, 1.38; P = .04) were associated with a higher risk of accident, but habitual visual acuity and myopia were unassociated with the crash risk, after adjusting for age, sex, time spent riding, and risky riding behaviors. CONCLUSION: These results may be consistent with data on motor vehicle accidents implicating peripheral vision (potentially compromised by spectacle wear) more strongly than central visual acuity in mediating crash risk.

Endothelial cell loss and surgically induced astigmatism after sutureless large-incision manual cataract extraction (SLIMCE).

OBJECTIVES:

To describe a modified manual cataract extraction technique, sutureless large-incision manual cataract extraction (SLIMCE), and to report its clinical outcomes.

METHODS:

Case notes of 50 consecutive patients with cataract surgery performed using the SLIMCE technique were retrospectively reviewed. Clinical outcomes 3 months after surgery were analyzed, including postoperative uncorrected visual acuity, best-corrected visual acuity, intraoperative and postoperative complications, endothelial cell loss, and surgically induced astigmatism using the vector analysis method.

RESULTS:

At the 3-month follow-up, all 50 patients had postoperative best-corrected visual acuity of at least 20/60, and 37 patients (74%) had visual acuity of at least 20/30. Uncorrected visual acuity was at least 20/68 in 28 patients (56%) and was between 20/80 and 20/200 in 22 patients (44%). No significant intraoperative complications were encountered, and sutureless wounds were achieved in all but 2 patients. At the 3-month follow-up, endothelial cell loss was 3.9%, and the mean surgically induced astigmatism was 0.69 diopter.

CONCLUSIONS:

SLIMCE is a safe and effective manual cataract extraction technique with low rates of surgically induced astigmatism and endothelial cell loss. In view of its low cost, SLIMCE may have a potential role in reducing cataract blindness in developing countries.

Hope Wounds: sistema de apoio à prática clínica na Diabetes Mellitus

Recentemente assiste-se a um crescimento da utilização das tecnologias de informação em diversas áreas, em particular dos dispositivos móveis. Na área da saúde é possível encontrar soluções inovadoras para apoio ao diagnóstico e tratamento de patologias que afectam milhares de pessoas em todo o mundo, como é o caso do risco de amputação em pacientes diabéticos. Aproveitando a existência de um ambiente de interligação telemática entre instituições de saúde Região do Baixo Vouga, o presente trabalho visa implementar o sistema HOPE Wounds na rotina diária de trabalho de profissionais de saúde, em especial médicos e enfermeiros, na prática da prestação de cuidados a pacientes diabéticos, com complicações de pé diabético. A interacção do dispositivo móvel e o ambiente web faz com que este sistema possibilite aceder e gravar diversa informação clínica, como por exemplo o registo fotográfico de feridas em escala temporal. O sistema HOPE Wounds integrado na Rede Telemática da Saúde permite ainda estabelecer a ponte entre profissionais do Centro de Saúde, da Consulta de Especialidade e do Internamento Hospitalar. A receptividade dos profissionais de saúde deixa antever a possibilidade de extensão a outras áreas médicas.

DNA damage in two bivalve species: Mytilus galloprovincialis and Ruditapes decussatus from the South coast of Portugal

Dissertação de mest., Estudos Marinhos e Costeiros, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2009

Managing Stress and Pain to Prevent Patient Discomfort, Distress and Delayed Wound Healing.

Evidence suggests stress slows the healing of wounds but pain may also play a part. Regular assessment could improve patients' quality of life and recovery time.

In vitro study of the effect of wound dressings on planktonic and biofilms from diabetic foot isolates

The management of wound bioburden has previously been evaluated using various antimicrobial wound dressings on bacterial pathogens isolated from various wounds. In this present study, the antimicrobial effect of silver-impregnated dressings (Acticoat and Silvercel) and honey-impregnated dressing (Medihoney™ Apinate) on both planktonic bacteria and quasi-biofilms by Staphylococcus aureus and Proteus mirabilis were assessed using a 6-well plate and standard agar technique. In the 6-well plate assay, a bacterial suspension of 108 colony forming unit (CFU)/mL was inoculated on each dressing in excess Luria-Bertani broth and incubated at 35 – 37°C for 30 and 60 minutes and 24 hours. After each incubation time, bacteria were recovered in sodium thioglycolate solution (STS) and the CFU/mL determined on LB agar. Dressings were cut into circular shapes (2cm diameter and placed on Mueller Hinton agar plates pre-inoculated with bacterial suspensions to determine their zones of inhibition (ZOI) after 24 hours incubation. None of the dressings was effective to significantly inhibit bacterial growth or biofilm formation at all the times tested. Acticoat and Medihoney™ Apinate produced ZOIs between 1.5 – 15 mm against both Staphylococcus aureus and Proteus mirabilis. It is possible that, dressings augmented with antibiotics can significantly reduce quasi-biofilms on standard agar.

In Vitro Assessment of the Synergy between Polymyxin B (PMB) and Polymyxin B Nonapeptide (PMBN) and Antibiotics on Biofilms from Diabetic Foot Infections

Background: The increasing resistance of Gram-negative bacteria isolated from nosocomial infections and chronic wounds, such as diabetic foot ulcers has renewed research interests in the use of polymyxins in the treatment of multidrug resistant infections. The added resistance conferred by biofilm development in such infections and the absence of novel antibiotics presuppose that polymyxins are the likely drugs of choice in spite of their nephrotoxicity. The effects of PMB and PMBN have been previously assessed on planktonic bacteria isolated from various infections. Methods: This current study assessed the synergy between a PMB/PMBN and two antibiotics (ceftazidime and levofloxacin) in an attempt to develop a strategy for biofilm disruption using the Minimum Biofilm Eradication Concentration Physiology and Genetic assay (MBEC™ P & G, Innovotech Inc, Edmonton, Alberta, Canada) according to manufacturer’s instructions. Klebsiella pneumoniae (K. pneumoniae) and Proteus mirabilis (P. mirabilis) biofilms of initial broth suspensions of 108 colony forming units per mL, cultivated on the pegs of the MBEC device were challenged with 5120 µg/mL of both ceftazidime and levofloxacin in a ten-fold dilution assay and in the presence of 100 and 500 µg/mL PMB and PMBN. Results: From table of results (Table 1), it can be deduced that both ceftazidime and levofloxacin are very effective in inhibiting biofilm development (as shown by percentage inhibition (PI)) when augmented with PMB and PMBN. This is about 100-fold increase in efficacy when compared to the antibiotics used on their own. The percentage reduction (PR) in biofilm was also increased considerably when PMB and PMBN concentrations were increased to 500 µg/mL. PMB was more effective than its less antibacterial derivative PMBN. Levofloxacin was also found to be more effective than ceftazidime when combined with both PMB and PMBN due to its enhanced cell-membrane permeability and as an anti-DNA replication uncoupling agent. Conclusion: The above results indicate that the synergy between antibiotics and cell membrane permeabilising agents may provide alternate strategies towards biofilm eradication

Investigating the rates and impacts of near misses and related incidents among UK cyclists

The paper investigates the occurrence of non-injury incidents among cyclists in the UK, seeking to (i) generate a rate that can be compared with injury rates, (ii) analyse factors affecting incident rates, and (iii) analyse factors affecting the impact of incidents on cyclists. We collected data on non-injury cycling ‘incidents’ (near misses and other frightening and/or annoying incidents) from 1692 online diaries of cycle trip stages1 and incidents, participants having signed up in advance for a specific day. Following data cleaning and coding, a dataset was created covering 1532 diary days and 3994 records of incidents occurring within the UK. Incident rates were calculated and compared to injury risks for cyclists. Cross-tabulation and regression were used to identify factors affecting incident rates and the effect an incident has on the cyclist. Frightening or annoying non-injury incidents, unlike slight injuries, are an everyday experience for most people cycling in the UK. For regular cyclists ‘very scary’ incidents (rated as 3 on a 0–3 scale) are on average a weekly experience, with deliberate aggression experienced monthly. Per mile, non-injury incidents were more frequent for people making shorter and slower trips. People aged over 55 were at lower risk, as were those cycling at the weekend and outside the morning peak. Incidents that involved motor vehicles, especially those involving larger vehicles, were more frightening than those that did not. Near miss and other non-injury incidents are widespread in the UK and may have a substantial impact on cycling experience and uptake. Policy and research should initially target the most frightening types of incident, such as very close passes and incidents involving large vehicles. Further attention needs to be paid to the experiences of groups under-represented among cyclists, such as women making shorter trips.

Application of geographic information system and modelling in health impact assessment for urban road mobility in Vietnam

Transport is an essential sector in modern societies. It connects economic sectors and industries. Next to its contribution to economic development and social interconnection, it also causes adverse impacts on the environment and results in health hazards. Transport is a major source of ground air pollution, especially in urban areas, and therefore contributing to the health problems, such as cardiovascular and respiratory diseases, cancer, and physical injuries. This thesis presents the results of a health risk assessment that quantifies the mortality and the diseases associated with particulate matter pollution resulting from urban road transport in Hai Phong City, Vietnam. The focus is on the integration of modelling and GIS approaches in the exposure analysis to increase the accuracy of the assessment and to produce timely and consistent assessment results. The modelling was done to estimate traffic conditions and concentrations of particulate matters based on geo-references data. A simplified health risk assessment was also done for Ha Noi based on monitoring data that allows a comparison of the results between the two cases. The results of the case studies show that health risk assessment based on modelling data can provide a much more detail results and allows assessing health impacts of different mobility development options at micro level. The use of modeling and GIS as a common platform for the integration of different assessments (environmental, health, socio-economic, etc.) provides various strengths, especially in capitalising on the available data stored in different units and forms and allows handling large amount of data. The use of models and GIS in a health risk assessment, from a decision making point of view, can reduce the processing/waiting time while providing a view at different scales: from micro scale (sections of a city) to a macro scale. It also helps visualising the links between air quality and health outcomes which is useful discussing different development options. However, a number of improvements can be made to further advance the integration. An improved integration programme of the data will facilitate the application of integrated models in policy-making. Data on mobility survey, environmental monitoring and measuring must be standardised and legalised. Various traffic models, together with emission and dispersion models, should be tested and more attention should be given to their uncertainty and sensitivity

Abordagem e maneio médico-cirúrgico de feridas abertas em cães e gatos : caracterização etiológica e estudo de padrões traumáticos

Dissertação de Mestrado Integrado em Medicina Veterinária

Nuclear factor I-C links platelet-derived growth factor and transforming growth factor beta1 signaling to skin wound healing progression.

Transforming growth factor beta (TGF-beta) and platelet-derived growth factor A (PDGFAlpha) play a central role in tissue morphogenesis and repair, but their interplay remain poorly understood. The nuclear factor I C (NFI-C) transcription factor has been implicated in TGF-beta signaling, extracellular matrix deposition, and skin appendage pathologies, but a potential role in skin morphogenesis or healing had not been assessed. To evaluate this possibility, we performed a global gene expression analysis in NFI-C(-/-) and wild-type embryonic primary murine fibroblasts. This indicated that NFI-C acts mostly to repress gene expression in response to TGF-beta1. Misregulated genes were prominently overrepresented by regulators of connective tissue inflammation and repair. In vivo skin healing revealed a faster inflammatory stage and wound closure in NFI-C(-/-) mice. Expression of PDGFA and PDGF-receptor alpha were increased in wounds of NFI-C(-/-) mice, explaining the early recruitment of macrophages and fibroblasts. Differentiation of fibroblasts to contractile myofibroblasts was also elevated, providing a rationale for faster wound closure. Taken together with the role of TGF-beta in myofibroblast differentiation, our results imply a central role of NFI-C in the interplay of the two signaling pathways and in regulation of the progression of tissue regeneration.

Bradykinin in blood and cerebrospinal fluid after acute cerebral lesions: correlations with cerebral edema and intracranial pressure.

Abstract Bradykinin (BK) was shown to stimulate the production of physiologically active metabolites, blood-brain barrier disruption, and brain edema. The aim of this prospective study was to measure BK concentrations in blood and cerebrospinal fluid (CSF) of patients with traumatic brain injury (TBI), subarachnoid hemorrhage (SAH), intracerebral hemorrhage (ICH), and ischemic stroke and to correlate BK levels with the extent of cerebral edema and intracranial pressure (ICP). Blood and CSF samples of 29 patients suffering from acute cerebral lesions (TBI, 7; SAH,: 10; ICH, 8; ischemic stroke, 4) were collected for up to 8 days after insult. Seven patients with lumbar drainage were used as controls. Edema (5-point scale), ICP, and the GCS (Glasgow Coma Score) at the time of sample withdrawal were correlated with BK concentrations. Though all plasma-BK samples were not significantly elevated, CSF-BK levels of all patients were significantly elevated in overall (n=73) and early (≤72 h) measurements (n=55; 4.3±6.9 and 5.6±8.9 fmol/mL), compared to 1.2±0.7 fmol/mL of controls (p=0.05 and 0.006). Within 72 h after ictus, patients suffering from TBI (p=0.01), ICH (p=0.001), and ischemic stroke (p=0.02) showed significant increases. CSF-BK concentrations correlated with extent of edema formation (r=0.53; p<0.001) and with ICP (r=0.49; p<0.001). Our results demonstrate that acute cerebral lesions are associated with increased CSF-BK levels. Especially after TBI, subarachnoid and intracerebral hemorrhage CSF-BK levels correlate with extent of edema evolution and ICP. BK-blocking agents may turn out to be effective remedies in brain injuries.

Predicting trabecular bone microdamage initiation and accumulation using a non-linear perfect damage model

Studies evaluating the mechanical behavior of the trabecular microstructure play an important role in our understanding of pathologies such as osteoporosis, and in increasing our understanding of bone fracture and bone adaptation. Understanding of such behavior in bone is important for predicting and providing early treatment of fractures. The objective of this study is to present a numerical model for studying the initiation and accumulation of trabecular bone microdamage in both the pre- and post-yield regions. A sub-region of human vertebral trabecular bone was analyzed using a uniformly loaded anatomically accurate microstructural three-dimensional finite element model. The evolution of trabecular bone microdamage was governed using a non-linear, modulus reduction, perfect damage approach derived from a generalized plasticity stress-strain law. The model introduced in this paper establishes a history of microdamage evolution in both the pre- and post-yield regions