Sequence Defined Disulfide-Linked Shuttle for Strongly Enhanced Intracellular Protein Delivery

Intracellular protein transduction technology is opening the door for a promising alternative to gene therapy. Techniques have to address all critical steps, like efficient cell uptake, endolysosomal escape, low toxicity, while maintaining full functional activity of the delivered protein. Here, we present the use of a chemically precise, structure defined three-arm cationic oligomer carrier molecule for protein delivery. This carrier of exact and low molecular weight combines good cellular uptake with efficient endosomal escape and low toxicity. The protein cargo is covalently attached by a bioreversible disulfide linkage. Murine 3T3 fibroblasts could be transduced very efficiently with cargo nlsEGFP, which was tagged with a nuclear localization signal. We could show subcellular delivery of the nlsEGFP to the nucleus, confirming cytosolic delivery and expected subsequent subcellular trafficking. Transfection efficiency was concentration-dependent in a directly linear mode and 20-fold higher in comparison with HIV-TAT-nlsEGFP containing a functional TAT transduction domain. Furthermore, β-galactosidase as a model enzyme cargo, modified with the carrier oligomer, was transduced into neuroblastoma cells in enzymatically active form

Influence of Permanent Magnet Characteristic Variability on the Wind Generator Operation

The paper discusses problems concerning the influence of permanent magnet material characteristics on the low-speed permanent magnet generator losses and output characteristics. The variability of the magnet material and its effect on the output parameters of the machine has been quantified. The characteristics of six different grades of neodymium permanent magnets have been measured and compared to the supplier specification data. The simulations of the generator have been carried out using transient finite element analysis. The results show that magnet materials from different suppliers have different characteristics, which have a significant influence on the generator output parameters, such as efficiency and power factor.Peer reviewe

Evaluation of transportan 10 in PEI mediated plasmid delivery assay.

Cell-penetrating peptides (CPPs) are novel high-capacity delivery vectors for different bioactive cargoes. We have evaluated the CPP transportan 10 (TP10) as a delivery vector in different in vitro plasmid delivery assays. Tested methods include: TP10 crosslinked to a plasmid via a peptide nucleic acid (PNA) oligomer, TP10 conjugation with polyethyleneimine (PEI), and addition of unconjugated TP10 to standard PEI transfection assay. We found that without additional DNA condensing agents, TP10 has poor transfection abilities. However, the presence of TP10 increases the transfection efficiency several folds compared to PEI alone. At as low concentrations as 0.6 nM, TP10-PNA constructs were found to enhance plasmid delivery up to 3.7-fold in Neuro-2a cells. Interestingly, the transfection efficiency was most significant at low PEI concentrations, allowing reduced PEI concentration without loss of gene delivery. No increase in cytotoxicity due to TP10 was observed and the uptake mechanism was determined to be endocytosis, as previously reported for PEI mediated transfection. In conclusion, TP10 can enhance PEI mediated transfection at relatively low concentrations and may help to develop future gene delivery systems with reduced toxicity

Adapting The Sniffin' Sticks Olfactory Test To Diagnose Parkinson's Disease In Estonia

The aim of the study was to develop a culturally adapted translation of the 12-item smell identification test from Sniffin' Sticks (SS-12) for the Estonian population in order to help diagnose Parkinson's disease (PD). Methods: A standard translation of the SS-12 was created and 150 healthy Estonians were questioned about the smells used as response options in the test. Unfamiliar smells were replaced by culturally familiar options. The adapted SS-12 was applied to 70 controls in all age groups, and thereafter to 50 PD patients and 50 age- and sex-matched controls. Results: 14 response options from 48 used in the SS-12 were replaced with familiar smells in an adapted version, in which the mean rate of correct response was 87% (range 73-99) compared to 83% with the literal translation (range 50-98). In PD patients, the average adapted SS-12 score (5.4/12) was significantly lower than in controls (average score 8.9/12), p<0.0001. A multiple linear regression using the score in the SS-12 as the outcome measure showed that diagnosis and age independently influenced the result of the SS-12. A logistic regression using the SS-12 and age as covariates showed that the SS-12 (but not age) correctly classified 79.0% of subjects into the PD and control category, using a cut-off of <7 gave a sensitivity of 76% and specificity of 86% for the diagnosis of PD. Conclusions: The developed SS-12 cultural adaption is appropriate for testing olfaction in Estonia for the purpose of PD diagnosis. © 2014 Elsevier Ltd.208830833Soudrya, Y., Lemogne, C., Malinvaud, D., Consoli, S.-M., Bonfils, P., Olfactory system and emotion: common substrates (2011) Eur Ann Otorhinolaryngol Head Neck Dis, 128, pp. 18-23Kovács, T., Mechanisms of olfactory dysfunction in aging and neurodegenerative disorders (2004) Ageing Res Rev, 3, pp. 215-232Chaudhuri, K.R., Odin, P., The challenge of non-motor symptoms in Parkinson's disease (2010) Prog Brain Res, 184, pp. 325-341Bohnen, N.I., Studenski, S.A., Constantine, G.M., Moore, R.Y., Diagnostic performance of clinical motor and non-motor tests of Parkinson disease: a matched case-control study (2008) Eur J Neurol, 15 (7), pp. 685-691Silveira-Moriyama, L., de Jesus Carvalho, M., Katzenschlager, R., Petrie, A., Ranvaud, R., Barbosa, E.R., The use of smell identification tests in the diagnosis of Parkinson's disease in Brazil (2008) Mov Disord, 23, pp. 2328-2334Silveira-Moriyama, L., Schwingenschuh, P., O'Donnell, A., Schneider, S.A., Mir, P., Carrillo, F., Olfaction in patients with suspected Parkinsonism and scans without evidence of dopaminergic deficit (SWEDDs) (2009) JNeurol Neurosurg Psychiatry, 80 (7), pp. 744-748Doty, R.L., Shaman, P., Kimmelman, C., Dann, M.S., University of Pennsylvania Smell Identification Test: a rapid quantitative olfactory function test for the clinic (1984) Laryngoscope, 94, pp. 176-178Doty, R.L., Marcus, A., Lee, W.W., Development of the 12-Item cross-cultural smell identification test (CC-SIT) (1996) Laryngoscope, 106, pp. 353-356Hummel, T., Konnerth, C.G., Rosenheim, K., Kobal, G., Screening of olfactory function using a 4 minute odor identification test: reliability, normative data, and investigations in patients with olfactory loss (2001) Ann Otol Rhinol Laryngol, 110, pp. 976-981Nordin, S., Bramerson, A., Liden, E., Bende, M., The Scandinavian odor-identification test: development, reliability, validity and normative data (1998) Acta Otolaryngol, 118, pp. 226-234Simmen, D., Briner, H.R., Hess, K., Screeningtest des Geruchssinnes mit Riechdiskette (1999) Laryngorhinootologie, 78, pp. 125-130Hummel, C., Zucco, G.M., Iannilli, E., Maboshe, W., Landis, B.N., Hummel, T., OLAF: standardization of international olfactory tests (2012) Eur Arch Otorhinolaryngol, 269 (3), pp. 871-880Cho, J.H., Jeong, Y.S., Lee, Y.J., Hong, S.C., Yoon, J.H., Kim, J.K., The Korean version of the Sniffin' stick (KVSS) test and its validity in comparison with the cross-cultural smell identification test (CC-SIT) (2009) Auris Nasus Larynx, 36, pp. 280-286Shu, C.H., Yuan, B.C., Lin, S.H., Lin, C.Z., Cross-cultural application of the "Sniffin' Sticks" odor identification test (2007) Am J Rhinol, 21 (5), pp. 570-573Silveira-Moriyama, L., Sirisena, D., Gamage, P., Ranjanie Gamage, R., Silva, R., Lees, A.J., Adapting the sniffin' sticks to diagnose PD in Sri Lanka (2009) Mov Disord, 24 (8), pp. 1229-1233Mackay-Sim, A., Grant, L., Owen, C., Chant, D., Silburn, P., Australian norms for a quantitative olfactory function test (2004) JClin Neurosci, 11, pp. 874-879Neumann, C., Tsioulos, K., Merkonidis, C., Salam, M., Clark, A., Philpott, C., Validation study of the "Sniffin' Sticks" olfactory test in a British population: a preliminary communication (2012) Clin Otolaryngol, 37 (1), pp. 23-27Eibenstein, A., Fioretti, A.B., Lena, C., Rosati, N., Ottaviano, I., Fusetti, M., Olfactory screening test: experience in 102 Italian subjects (2005) Acta Otorhinolaryngol Ital, 25 (1), pp. 18-22Boesveldt, S., Verbaan, D., Knol, D.L., Visser, M., van Rooden, S.M., van Hilten, J.J., Acomparative study of odor identification and odor discrimination deficits in Parkinson's disease (2008) Mov Disord, 23 (14), pp. 1984-1990Boesveldt, S., de Muinck Keizer, R.J., Knol, D.L., Wolters, E., Berendse, H.W., Extended testing across, not within, tasks raises diagnostic accuracy of smell testing in Parkinson's disease (2009) Mov Disord, 24 (1), pp. 85-90Hummel, T., Kobal, G., Gudziol, H., Mackay-Sim, A., Normative data for the "Sniffin' Sticks" including tests of odor identification, odor discrimination, and olfactory thresholds: an upgrade based on a group of more than 3,000 subjects (2007) Eur Arch Otorhinolaryngol, 264, pp. 237-243Doty, R.L., Olfactory dysfunction in Parkinson disease (2012) Nat Rev Neurol, 8 (6), pp. 329-339Brand, G., Millot, J.L., Sex differences in human olfaction: between evidence and enigma (2001) QJ Exp Psychol B, 54 (3), pp. 259-270Larsson, M., Nilsson, L.G., Olofsson, J.K., Nordin, S., Demographic and cognitive predictors of cued odor identification: evidence from a population-based study (2004) Chem Senses, 29 (6), pp. 547-554Doty, R.L., Cameron, E.L., Sex differences and reproductive hormone influences on human odor perception (2009) Physiol Behav, 97 (2), pp. 213-228Orhan, K.S., Karabulut, B., Keles, N., Deger, K., Evaluation of factors concerning the olfaction using the sniffin' sticks test (2012) Otolaryngol Head Neck Surg, 146 (2), pp. 240-246Frye, R.E., Schwartz, B.S., Doty, R.L., Dose-related effects of cigarette smoking on olfactory function (1990) JAMA, 263, pp. 1233-1236Katotomichelakis, M., Balatsouras, D., Tripsianis, G., Davris, S., Maroudias, N., Danielides, V., The effect of smoking on the olfactory function (2007) Rhinology, 45, pp. 273-280Bower, J.H., Maraganore, D.M., Peterson, B.J., Ahlskog, J.E., Rocca, W.A., Immunologic diseases, anti-inflammatory drugs, & Parkinson disease: a case-control study (2006) Neurology, 67 (3), pp. 494-496Sedig, L., Leibner, J., Ramjit, A.L., Wu, S.S., Dai, Y., Okun, M.S., Is rhinorrhea an under-recognized intrinsic symptom of Parkinson disease? A prospective pilot study (2010) Int J Neurosci, 120 (4), pp. 258-26

A new fiber-chip coupling concept for reusable multipurpose packaging suitable for up to 45 GHz bandwidth

We designed a series of modules for fiber-chip coupling of InP-based OEICs, which are well suited for small-scale production. Simultaneous coupling of both chip sides to fibers is possible with a new patented set-up