Proteomic analysis reveals mechanisms underlying increased efficacy of bleomycin by photochemical internalization in bladder cancer cells

Photochemical internalization (PCI) is a promising new technology for site-specific drug delivery, developed from photodynamic therapy (PDT). In PCI, light-induced activation of a photosensitizer trapped inside endosomes together with e.g. chemotherapeutics, nucleic acids or immunotoxins, allows cytosolic delivery and enhanced local therapeutic effect. Here we have evaluated the photosensitizer meso-tetraphenyl chlorine disulphonate (TPCS2a/fimaporfin) in a proteome analysis of AY-27 rat bladder cancer cells in combination with the chemotherapeutic drug bleomycin (BML). We find that BLMPCI attenuates oxidative stress responses induced by BLM alone, while concomitantly increasing transcriptional repression and DNA damage responses. BLMPCI also mediates downregulation of bleomycin hydrolase (Blmh), which is responsible for cellular degradation of BLM, as well as several factors known to be involved in fibrotic responses. PCI-mediated delivery might thus allow reduced dosage of BLM and alleviate unwanted side effects from treatment, including pulmonary fibrosis.publishedVersionPaid open acces

δ-Aminolevulinic acid transport in murine mammary adenocarcinoma cells is mediated by beta transporters

δ-aminolevulinic acid, the precursor of porphyrin biosynthesis has been used to induce the endogenous synthesis of the photosensitiser protoporphyrin IX for photodynamic therapy in the treatment of various tumours. The aim of this work was to characterise the δ-aminolevulinic acid transport system in the murine mammary adenocarcinoma cell line LM3 using 14C-δ-aminolevulinic acid, to finally improve δ-aminolevulinic acid incorporation in mammalian cells. Our results showed that δ-aminolevulinic acid is incorporated into these cells by two different mechanisms, passive diffusion which is important at the beginning of the incubation, and active transport. Specificity assays suggested that the transporter involved in δ-aminolevulinic acid incorporation is a BETA transporter, probably GAT-2

Developing a framework of minimum standards for the risk assessment of alien species

1. Biological invasions are a threat to biodiversity, society and the economy. There is an urgent need to provide evidence- based assessments of the risks posed by invasive alien species (IAS) to prioritize action. Risk assessments underpin IAS policies in many ways: informing legislation; providing justification of restrictions in trade or consumer activities; prioritizing surveillance and rapid response. There are benefits to ensuring consistency in content of IAS risk assessments globally, and this can be achieved by providing a framework of minimum standards as a checklist for quality assurance. 2. From a review of existing risk assessment protocols, and with reference to the requirements of the EU Regulation on IAS (1143/2014) and international agreements including the World Trade Organisation, Convention on Biological Diversity and International Plant Protection Convention, coupled with consensus methods, we identified and agreed upon 14 minimum standards (attributes) a riskassessment scheme should include. 3. The agreed minimum standards were as follows: (1) basic species description; (2) likelihood of invasion; (3) distribution, spread and impacts; (4) assessment of introduction pathways; (5) assessment of impacts on biodiversity and ecosystems; (6) Assessment of impact on ecosystem services; (7) assessment of socio-economic impacts; (8) consideration of status (threatened or protected) of species or habitat under threat; (9) assessment of effects of future climate change; (10) completion possible even when there is a lack of information; (11) documents information sources; (12) provides a summary in a consistent and interpretable form; (13) -includes uncertainty; (14) includes quality assurance. In deriving these minimum standards, gaps in knowledge required for completing risk assessments and the scope of existing risk assessment protocols were revealed, most notably in relation to assessing benefits, socio-economic impacts and impacts on ecosystem services but also inclusion of consideration of climate change. 4. Policy implications. We provide a checklist of components that should be within invasive alien species risk assessments and recommendations to develop risk assessments to meet these proposed minimum standards. Although inspired by implementation of the European Union Regulation on invasive alien species, and as such developed specifically within a European context, the derived framework and minimum standards could be applied globally

Porphyrin accumulation induced by 5-aminolaevulinic acid esters in tumour cells growing in vitro and in vivo

The ability of 5-aminolaevulinic acid and some of its esterified derivatives to induce porphyrin accumulation has been examined in CaNT murine mammary carcinoma cells growing in culture and as tumours in vivo. Topical or intravenous administration of 5-aminolaevulinic acid-esters to mice bearing subcutaneous tumours produced lower porphyrin levels in the tumour than an equimolar dose of 5-aminolaevulinic acid. Reducing the dose of intravenous hexyl- or benzyl-ALA and topical hexyl-5-aminolaevulinic acid resulted in a dose-dependent reduction in porphyrin accumulation. A number of normal tissues accumulated higher concentrations of porphyrins than tumour tissue following intravenous administration of 5-aminolaevulinic acid-esters. Esterase activity in these normal tissues was greater than that in tumour tissue. In contrast to the situation in vivo, all of the 5-aminolaevulinic acid-esters examined were at least as effective as 5-aminolaevulinic acid when applied to cloned CaNT cells in vitro, with the drug concentration required for maximum porphyrin accumulation varying with ester chain-length. Tumour cells growing in culture released esterase activity into the medium. These findings suggest that the efficacy of 5-aminolaevulinic esters may vary depending on the esterase activity of the target tissue, and suggest caution when interpreting the findings of in vitro studies using these and similar prodrugs

Methotrexate used in combination with aminolaevulinic acid for photodynamic killing of prostate cancer cells

Photodynamic therapy (PDT) using 5-aminolaevulinic acid (ALA) to drive production of an intracellular photosensitiser, protoporphyrin IX (PpIX), is a promising cancer treatment. However, ALA-PDT is still suboptimal for thick or refractory tumours. Searching for new approaches, we tested a known inducer of cellular differentiation, methotrexate (MTX), in combination with ALA-PDT in LNCaP cells. Methotrexate alone promoted growth arrest, differentiation, and apoptosis. Methotrexate pretreatment (1 mg l−1, 72 h) followed by ALA (0.3 mM, 4 h) resulted in a three-fold increase in intracellular PpIX, by biochemical and confocal analyses. After exposure to 512 nm light, killing was significantly enhanced in MTX-preconditioned cells. The reverse order of treatments, ALA-PDT followed by MTX, yielded no enhancement. Methotrexate caused a similar relative increase in PpIX, whether cells were incubated with ALA, methyl-ALA, or hexyl-ALA, arguing against a major effect upon ALA transport. Searching for an effect among porphyrin synthetic enzymes, we found that coproporphyrinogen oxidase (CPO) was increased three-fold by MTX at the mRNA and protein levels. Transfection of LNCaP cells with a CPO-expressing vector stimulated the accumulation of PpIX. Our data suggest that MTX, when used to modulate intracellular production of endogenous PpIX, may provide a new combination PDT approach for certain cancers

Biological flora of Central Europe: Cyperus esculentus L

This paper presents information on all aspects of the biology of Cyperus esculentus L. (yellow nutsedge) and deals with its taxonomy, morphology, genetic diversity, distribution, habitat requirements, ecology and life cycle, with special emphasis on uses and cultivation, history of introduction, impact and management in Europe. C. esculentus is a tuber geophyte and most likely originates from the Mediterranean and Southwest Asia. It is a variable plant and four wild-type varieties are presently recognized, in addition to a cultivated form. C. esculentus reproduces primarily by its underground tubers, although abundant seeds are produced. In temperate climates, tubers usually sprout in late spring and the plant withers at the beginning of the winter. C. esculentus is only cultivated in the València region in Spain. Invasion foci emerged across Europe at the beginning of the 1980s and at present, C. esculentus is most abundant on arable land and in ruderal habitats, followed by riverine vegetation. In heavily infested regions of Europe, C. esculentus causes substantial yield losses in field crops and although different management strategies are available, C. esculentus remains difficult to control.Follak, S.; Belz, R.; Bohren, C.; Castro, OD.; Guacchio, ED.; Pascual-Seva, N.; Schwarz, M.... (2016). Biological flora of Central Europe: Cyperus esculentus L. Perspectives in Plant Ecology, Evolution and Systematics. 23:33-51. doi:10.1016/j.ppees.2016.09.003S33512

Ortodontiskā ārstēšana pacientiem ar periodontālām saslimšanām

ZobārstniecībaVeselības aprūpeDentistryHealth CareMūsdienās gan jauniešu, gan pieaugušo populācijas uzmanības centrā arvien vairāk nokļūst dentofaciālā estētika. Pieaugušo pacientu skaits, kas vēlas izlīdzināt zobus ar ortodontiskās ārstēšanas palīdzību, pēdējo 2-3 desmitgažu laikā, ir palielinājies. Novecošanās izsaukto periodontālo audu izmaiņu dēļ, daudziem no šiem pacientiem ir kompromitēta periodontālā veselība. Lai izvairītos no tālākas periodontālo audu destrukcijas, jebkurš aktīvs periodontālo audu iekaisums ir jānovērš pirms ortodontisko aparatūru uzlikšanas. Fiksētās ortodontiskās breketes akumulē vairāk zobu aplikumu un baktērijas, līdz ar to pacientam obligāti ir nepieciešamas regulāras kontroles un uzturošās ārstēšanas vizītes, kurās tiktu veikta mutes dobuma higiēna un zobu aplikuma kontrole. Tā kā zobu aplikums un baktēriju akumulācija, ja tā netiek novērsta, var izraisīt tālāku periodontālo audu destrukciju, tad pacienta sadarbība ir ļoti svarīga. Šis pētnieciskais darbs apskata ortodontiskās ārstēšanas iespējas pacientiem ar kompromitētu periodontu.Rezumējot - pacientiem ar kompromitētu periodontālo veselību ortodontiskā ārstēšana ir iespējama, ja tiek ievēroti vairāki priekšnosacījumi un, ja pacients ir ne tikai labi informēts par mutes dobuma higiēnu, bet arī ļoti labi motivēts.Dentofacial aesthetics get more and more focus in daily life nowadays, for both the younger population and the adult population. The last 2-3 decades the number of adult patients who wants to align teeth with orthodontic treatment has increased, and due to aging changes in the periodontium many of them have compromised periodontal health. In order to avoid further destruction of the periodontium, any active periodontal inflammation needs to be eliminated before placement of orthodontic appliances. As fixed orthodontic brackets will accumulate more dental plaque and bacteria, regular follow-up maintenance with oral hygiene treatment and strict plaque control with the patient is mandatory. Patient compliance is essential, as dental plaque and bacteria accumulation will lead to further destruction of the periodontium if not handled. This paper will look at the relationship of orthodontic treatment in patients who already have a compromised periodontium. In short, orthodontic treatment in patients with compromised periodontal health is possible if several precautions are followed, and the patient is well educated and motivated about oral hygiene maintenance

Test retest reliability of computer-based video analysis for the assessment of postural control in individuals with cerebral palsy

BACKGROUND: Deficits in the control of posture is an important aspect of CP, as defined by SCPE and children with CP often exhibit activity limitations that are associated with postural control problems. However, there is a lack of adequate tools for assessing postural control in individuals with CP, and there is limited documentation of the measurement properties of the existing tools. AIM: The first aim of this study was to explore if variables obtained using a computer-based video analysis software could be used to assess postural control. The second aim of this student thesis was to explore the test-retest reliability properties of these variables in individuals with CP and TD individuals 8-29 years of age. METHOD: Thirteen individuals with CP and 24 typically developing (TD) individuals, 8-29 years, were asked to stand still while three videos recorded their movements from the side. The movements in the video recordings were quantified, using a computer-based video analysis. RESULTS: In the explorative part of this study we found that the variables calculated by the computer-based video analysis that best correlated with the clinical assessment of postural control in the individuals with CP was the mean value of the centroid of motion (Cxmean) and its standard deviation (CxSD) in the horizontal axis. The centroid of motion is the spatial centre of all movements in the picture. In further analyses, the Cxmean showed the best correlations with GMFM-66, a clinical assessment tool for postural control, while the correlations were low for CxSD. In the second part, I found that when we included all participants in the analyses, the ICC values of Cxmean ranged between 0.89 and 0.93, and of CxSD ranged between 0.92 and 0.93. The ICC values of Cxmean and CxSD were higher with more narrow confidence intervals when two video recordings, each of 30 seconds duration, were included, than when a third video recording of two minutes was included in the calculations. The ICC values of Cxmean and CxSD were nearly identical when applying ICC(1,1) and ICC(3,1). The standard error of measurement (SEM) for Cxmean ranged from 2.2 (4 %) to 3.1 (6 %), expressing a small degree of measurement error. The smallest detectable difference (SDD) for Cxmean ranged from 6.0 (10 %) to 8.5 (15 %). However, the SEM values for CxSD ranged from 0.3 (14 %) to 0.7 (27 %), and the SDD values from 0.7 (40 %) to 2.0 (76 %). The Bland-Altman plots for Cxmean verifies graphically the consistency of the 3 video recordings. The Bland-Altman plots for CxSD verifies graphically the consistency of the measures in TD group, while it illustrates a spread in the values in the individuals with CP. The difference between the CxSD values from the two recordings were larger in the individuals with CP than in those with TD. CONCLUSION: We found that the variable Cxmean from the computer-based video analysis software describes certain aspects of postural control in individuals with CP, 8-29 years. The test-retest reliability of this variable is good. However, more studies are required to further evaluate this method and to explore if other variables of the computer-based video analysis may better describe other aspects of postural control