Mechanisms for the Intracellular Manipulation of Organelles by Conventional Electroporation

Conventional electroporation (EP) changes both the conductance and molecular permeability of the plasma membrane (PM) of cells and is a standard method for delivering both biologically active and probe molecules of a wide range of sizes into cells. However, the underlying mechanisms at the molecular and cellular levels remain controversial. Here we introduce a mathematical cell model that contains representative organelles (nucleus, endoplasmic reticulum, mitochondria) and includes a dynamic EP model, which describes formation, expansion, contraction, and destruction for the plasma and all organelle membranes. We show that conventional EP provides transient electrical pathways into the cell, sufficient to create significant intracellular fields. This emerging intracellular electrical field is a secondary effect due to EP and can cause transmembrane voltages at the organelles, which are large enough and long enough to gate organelle channels, and even sufficient, at some field strengths, for the poration of organelle membranes. This suggests an alternative to nanosecond pulsed electric fields for intracellular manipulations.National Science Foundation (U.S.) (NSF Graduate Research Fellowship)National Institutes of Health (U.S.) (grant No. R01-GM63857)Aegis Industries, Inc

Enhanced Collateral Growth by Double Transplantation of Gene-Nucleofected Fibroblasts in Ischemic Hindlimb of Rats

BACKGROUND: Induction of neovascularization by releasing therapeutic growth factors is a promising application of cell-based gene therapy to treat ischemia-related problems. In the present study, we have developed a new strategy based on nucleofection with alternative solution and cuvette to promote collateral growth and re-establishment of circulation in ischemic limbs using double transplantation of gene nucleofected primary cultures of fibroblasts, which were isolated from rat receiving such therapy. METHODS AND RESULTS: Rat dermal fibroblasts were nucleofected ex vivo to release bFGF or VEGF165 in a hindlimb ischemia model in vivo. After femoral artery ligation, gene-modified cells were injected intramuscularly. One week post injection, local confined plasmid expression and transient distributions of the plasmids in other organs were detected by quantitative PCR. Quantitative micro-CT analyses showed improvements of vascularization in the ischemic zone (No. of collateral vessels via micro CT: 6.8±2.3 vs. 10.1±2.6; p<0.05). Moreover, improved collateral proliferation (BrdU incorporation: 0.48±0.05 vs. 0.57±0.05; p<0.05) and increase in blood perfusion (microspheres ratio: gastrocnemius: 0.41±0.10 vs. 0.50±0.11; p<0.05; soleus ratio: soleus: 0.42±0.08 vs. 0.60±0.08; p<0.01) in the lower hindlimb were also observed. CONCLUSIONS: These results demonstrate the feasibility and effectiveness of double transplantation of gene nucleofected primary fibroblasts in producing growth factors and promoting the formation of collateral circulation in ischemic hindlimb, suggesting that isolation and preparation of gene nucleofected cells from individual accepting gene therapy may be an alternative strategy for treating limb ischemia related diseases

Development of Lithuanian cartography (since earliest times until the middle of the 20th century)

W artykule omówiono cztery okresy rozwoju kartografii Litwy od czasów antycznych do pierwszej połowy XX wieku, uwzględniając szczególnie powstające w ostatnim okresie mapy tematyczne.The first cartographic views of Lithuanian territory appeared in antique times. Regular mapping of the territory started in 15th century. The long (over 400 years) period until the first half of 20th century can be naturally divided into 4 periods, every of which can be characterized by the specifics of cartographic representations for the period: thematic and contents of maps, ways and means of cartographic visualization, amount and quality of the represented data. The very first period of Lithuanian cartography development can be given a name of the non-professional period. Cartographic issues were created by people without special education. Nevertheless, many informative and quite accurate maps were designed during this period: Sarmatia in terra Europa by Nicolaus Cusanus (1491), Mapa regni Poloniae et Magni Ducatus Lithuaniae by Bernard Wapowski (1526), Magni Ducatus Lithuaniae, Livoniae et Moscoviae descripto by Maciej Strubicz (1589), Poloniae finimarumque locorum descriptio by Wacław Grodecki (1570). The most outstanding map of this period is the Magni Ducatus Lithuaniae, caeterarumque regionum illi adiacentium exacta descriptio by Kristupas Radvila and Tomasz Makowski (1613). The second period relates to the formation of the basics of theoretical cartography. Vilnius Academy (University) was established in 1579 but natural science was not taught in it yet. Only in early 18th century the astronomy and geodesy lecture courses were given. In the second half of 18th century general and physical geography was already taught in Vilnius University (by Karol Wyrwycz), and the astronomic observatory established. At that time first geography manuals were written by the professors of the University (K. Hołowka, 1743, F. Paprocki, 1754, K. Wyrwicz, 1768, 1770). These manuals contains also the basics of geodetic survey and elements of cartography. In 17th and 18th centuries several general maps of Lithuania were designed: Partie de Lithuanie... (1665), La Curlande Duche et Semigale...(1659) by Nicolas Sanson d'Abbeville, Magni Ducatus Lithuania...by F. de Wit (1680), Magni Ducatus Lithuania... by J. Nieprzecki (1749). In late 18th century as a result of undertaken local topographical survey, city plants of Vilnius, Taurage, Kedainiai, Svedasai were drawn. At the very end of 18th century the professor of Vilnius university S.B. Jumdzilas compiled the geobotanic map of Gardinas environs, few years later such a map for Vilnius environs was made by professor Emanuel Gilibert. The period of professional topography began in early 19th century and lasted for almost one hundred years. In 1820 the department of Geodesy was founded in Vilnius University. The specialists and postgraduate students of this department took part in building the triangulation network for Lithuania and topographic survey of Lithuanian towns. The university professor Jan Śniadecki prepared the manual of topography that was for long time used in higher educational establishments of the Russian Empire. In early 19th century professor Joachim Lelewel made an abundant collection of old maps and atlases that became the core of the cartographic collection of the university. In the second half of 19th century during the survey of Lithuania the maps st scale 1:84 000, 1:42 000 and 1:21 000 were compiled. The period of thamatic cartography began in the late 19th century and lasted until the middle of 20th century. In 1876 the first map in Lithuanian language was published. The first geological and geomorphological maps were also compiled in 19th century. The first map in Lithuanian language for the territory of Lithuania was compiled by A. Macijauskas in 1900. Before the World War I cities plans were compiled for Vilnius, Kaunas, Siauliai, Panevetys, Ukmerge. During the interwar thematical cartography gained its pace. Geological map (J. Dalinkevicius, 1928, 1935, 1939), maps of natural resources (M. Kaveckis, 1939), geomorphological map (C. Pakuckas, 1939), map of the soils of Southem Lithuania (V. Ruokis, 1936), archaeological map (P. Tarasenka, 1928), historical maps (O. Maksimaitiene, 1936, 1937), and maps for schools (A. Vireliunas, 1930, P. Sinkunas, 1938) were compiled. In 1923 instrumental topographic survey of Lithuanian territory began. Until 1938 the topographic maps at scale 1:100 000 were compiled for the whole territory together with 92 sheets at scale 1:25 000. The intense development of cartography was interrupted by the rage of the Wprld War II

Semiotyczna ocena litewskich wojskowych map nawigacji powietrznej

Research of semiotic aspects Lithuanian military air navigation charts was based on the semantic, graphic and information load analysis. The aim of semantic analysis was to determine how the conventional cartographical symbols, used in air navigation charts, correspond with carto-linguistic and carto-semiotic requirements. The analysis of all the markings was performed complex and collected by questionnaire were interviewed various respondents: pilots, cartographers and other chart users. The researches seek two aims: evaluate information and graphical load of military air navigation charts. Information load evaluated to calculate all objects and phenomenon, which was in 25 cm² of map. Charts analysis showed that in low flight charts (LFC) average information load are 4 - 5 times richer than in the operational maps. Map signs optimization on LFC has to be managed very carefully, choosing signs that can reduce the load of information and helps for the information transfer process. Graphical load of maps evaluated of aeronautical maps is not great (5 - 12%) and does not require reduction the information load and generalization of charts. Air navigation charts analysis pointed that not all air navigation sings correspond carto-semiotic requirements and must be improved. The authors suggested some new sings for military air navigation chart, which are simpler, equivalent to human psychophysical perception criteria, creates faster communication and less load on the chart.Badania semiotycznych aspektów litewskich wojskowych map żeglugi powietrznej bazowały na semantycznej, graficznej i informacyjnej analizie treści tych map. Celem analizy semantycznej było określenie na ile tradycyjne symbole kartograficzne stosowane na mapach nawigacji lotniczej są zgodne z wymogami języka kartograficznego oraz zasadami stosowania znaków kartograficznych. Powyższe analizy przeprowadzono w sposób kompleksowy, informacje zebrano za pomocą ankiet, przeprowadzając wywiady w różnych środowiskach, w tym m. in. wśród pilotów, kartografów oraz innych użytkowników map. Prowadzone badania miały dwa podstawowe cele: ocena informacyjnego oraz graficznego wypełnienia treścią wojskowych map żeglugi powietrznej. Wypełnienie mapy informacjami oceniono zliczając wszystkie obiekty i zjawiska znajdujące się na obszarze 25 cm² mapy. Analiza ta wykazała, że mapy dla lotów na niskich pułapach (LFC) posiada ją średnio 4-5 razy większy zasób informacji niż mapy operacyjne. Zatem dobór znaków na mapie LFC powinien być wykonywany bardzo starannie, poprzez wybór takich znaków, które mogą zmniejszyć nasycenie mapy informacjami jednocześnie ułatwiając proces ich przekazywania. Nasycenie graficzne ocenianych map lotniczych nie jest zbyt duże (5-12%) i nie wymaga ograniczenia ilości przedstawianych informacji ani generalizacji. Analiza map żeglugi powietrznej wykazała, że nie wszystkie znaki występujące na tego rodzaju mapach spełniają wymagania dotyczące stosowania znaków kartograficznych i powinny być one udoskonalone. Autorzy zaproponowali kilka nowych znaków dla wojskowych map żeglugi powietrznej - są one prostsze, odpowiadają psychofizycznym możliwościom percepcyjnym człowieka, sprzyjają szybszej komunikacji oraz zmniejszają nasycenie informacyjne map

Semiotic evaluation of Lithuania military air navigation charts

Research of semiotic aspects Lithuanian military air navigation charts was based on the semantic, graphic and information load analysis. The aim of semantic analysis was to determine how the conventional cartographical symbols, used in air navigation charts, correspond with carto-linguistic and carto-semiotic requirements. The analysis of all the markings was performed complex and collected by questionnaire were interviewed various respondents: pilots, cartographers and other chart users. The researches seek two aims: evaluate information and graphical load of military air navigation charts. Information load evaluated to calculate all objects and phenomenon, which was in 25 cm² of map. Charts analysis showed that in low flight charts (LFC) average information load are 4 - 5 times richer than in the operational maps. Map signs optimization on LFC has to be managed very carefully, choosing signs that can reduce the load of information and helps for the information transfer process. Graphical load of maps evaluated of aeronautical maps is not great (5 - 12%) and does not require reduction the information load and generalization of charts. Air navigation charts analysis pointed that not all air navigation sings correspond carto-semiotic requirements and must be improved. The authors suggested some new sings for military air navigation chart, which are simpler, equivalent to human psychophysical perception criteria, creates faster communication and less load on the chart.Badania semiotycznych aspektów litewskich wojskowych map żeglugi powietrznej bazowały na semantycznej, graficznej i informacyjnej analizie treści tych map. Celem analizy semantycznej było określenie na ile tradycyjne symbole kartograficzne stosowane na mapach nawigacji lotniczej są zgodne z wymogami języka kartograficznego oraz zasadami stosowania znaków kartograficznych. Powyższe analizy przeprowadzono w sposób kompleksowy, informacje zebrano za pomocą ankiet, przeprowadzając wywiady w różnych środowiskach, w tym m. in. wśród pilotów, kartografów oraz innych użytkowników map. Prowadzone badania miały dwa podstawowe cele: ocena informacyjnego oraz graficznego wypełnienia treścią wojskowych map żeglugi powietrznej. Wypełnienie mapy informacjami oceniono zliczając wszystkie obiekty i zjawiska znajdujące się na obszarze 25 cm² mapy. Analiza ta wykazała, że mapy dla lotów na niskich pułapach (LFC) posiada ją średnio 4-5 razy większy zasób informacji niż mapy operacyjne. Zatem dobór znaków na mapie LFC powinien być wykonywany bardzo starannie, poprzez wybór takich znaków, które mogą zmniejszyć nasycenie mapy informacjami jednocześnie ułatwiając proces ich przekazywania. Nasycenie graficzne ocenianych map lotniczych nie jest zbyt duże (5-12%) i nie wymaga ograniczenia ilości przedstawianych informacji ani generalizacji. Analiza map żeglugi powietrznej wykazała, że nie wszystkie znaki występujące na tego rodzaju mapach spełniają wymagania dotyczące stosowania znaków kartograficznych i powinny być one udoskonalone. Autorzy zaproponowali kilka nowych znaków dla wojskowych map żeglugi powietrznej - są one prostsze, odpowiadają psychofizycznym możliwościom percepcyjnym człowieka, sprzyjają szybszej komunikacji oraz zmniejszają nasycenie informacyjne map

Bioregulatory Systems Medicine: An Innovative Approach to Integrating the Science of Molecular Networks, Inflammation and Systems Biology with the Patient’s Autoregulatory Capacity?

Bioregulatory systems medicine is a paradigm that aims to advance current medical practices. The basic scientific and clinical tenets of this approach embrace an interconnected picture of human health, supported largely by recent advances in systems biology and genomics, and focuses on the implications of multi-scale interconnectivity for improving therapeutic approaches to disease. This article introduces the formal incorporation of these scientific and clinical elements into a cohesive theoretical model of the bioregulatory systems medicine approach. The authors review this integrated body of knowledge and discuss how the emergent conceptual model offers the medical field a new avenue for extending the armamentarium of current treatment and healthcare, with the ultimate goal of improving population health