A novel three-dimensional culture system for isolation and clonal propagation of neural stem cells using a thermo-reversible gelation polymer

In the present study, we examined the possible utility of a three-dimensional culture system using a thermo-reversible gelation polymer to isolate and expand neural stem cells (NSCs). The polymer is a synthetic biologically inert polymer and gelates at temperatures higher than the gel-sol transition point ( approximately 20 degrees C). When fetal mouse brain cells were inoculated into the gel, spherical colonies were formed ( approximately 1% in primary culture and approximately 9% in passage cultures). The spheroid-forming cells were positive for expression of the NSC markers nestin and Musashi. Under conditions facilitating spontaneous neural differentiation, the spheroid-forming cells expressed genes characteristic to astrocytes, oligodendrocytes, and neurons. The cells could be successively propagated at least to 80 poly-D-lysines over a period of 20 weeks in the gel culture with a growth rate higher than that observed in suspension culture. The spheroids formed by fetal mouse brain cells in the gel were shown to be of clonal origin. These results indicate that the spheroid culture system is a convenient and powerful tool for isolation and clonal expansion of NSCs in vitro.

Numerical study of film cooling scheme on a blunt-nosed body in hypersonic flow

In hypersonic flights, the prediction of aerodynamic heating and the construction of a proper thermal protection system (TPS) are significantly important. In this study, the method of a film cooling technique, which is already the state of the art in cooling gas turbine engine, is proposed for a fully reusable and active TPS. Effectiveness of the film cooling scheme to reduce convective heating rates for a blunt nosed spacecraft flying at Mach number 6.56 and 40 degree angle of attack is investigated numerically. The inflow boundary conditions used the standard values at an altitude of 30 km. Computational domain consists of infinite rows of film cooling holes on the bottom of a blunt-nosed slab. Laminar and several turbulent calculations have been performed and compared each other. The influence of blowing ratios on the film cooling effectiveness is investigated. The results exhibit that the film cooling technique could be an effective method for an active cooling of blunt-nosed bodies in hypersonic flows.

Hydrogel-forming microneedle arrays exhibit antimicrobial properties:potential for enhanced patient safety

We describe, for the first time, the microbial characterisation of hydrogel-forming polymeric microneedle arrays and the potential for passage of microorganisms into skin following microneedle penetration. Uniquely, we also present insights into the storage stability of these hydroscopic formulations, from physical and microbiological viewpoints, and examine clinical performance and safety in human volunteers. Experiments employing excised porcine skin and radiolabelled microorganisms showed that microorganisms can penetrate skin beyond the stratum corneum following microneedle puncture. Indeed, the numbers of microorganisms crossing the stratum corneum following microneedle puncture were greater than 105 cfu in each case. However, no microorganisms crossed the epidermal skin. When using a 21G hypodermic needle, more than 104 microorganisms penetrated into the viable tissue and 106 cfu of Candida albicans and Staphylococcus epidermidis completely crossed the epidermal skin in 24 h. The hydrogel-forming materials contained no microorganisms following de-moulding and exhibited no microbial growth during storage, while also maintaining their mechanical strength, apart from when stored at relative humidities of 86%. No microbial penetration through the swelling microneedles was detectable, while human volunteer studies confirmed that skin or systemic infection is highly unlikely when polymeric microneedles are used for transdermal drug delivery. Since no pharmacopoeial standards currently exist for microneedle-based products, the exact requirements for a proprietary product based on hydrogel-forming microneedles are at present unclear. However, we are currently working towards a comprehensive specification set for this microneedle system that may inform future developments in this regard.

Two-dimensional images of dissolved sulfide and metals in anoxic sediments by a novel diffusive gradients in thin film probe and optical scanning techniques

A novel diffusive gradients in thin film probe developed comprises diffusive gel layer of silver iodide (AgI) and a back-up Microchelex resin gel layer. 2D high-resolution images of sulfide and trace metals were determined respectively on the AgI gel by densitometric analysis and on the Microchelex resin layer with laser-ablation-inductively-coupled plasma mass spectrometry (LA-ICP-MS).We investigated the validity of the analytical procedures used for the determination of sulfide and trace metals. We found low relative standard deviations on replicate measurements, linear trace-metal calibration curves between the LA-ICP-MS signal and the true trace-metal concentration in the resin gel, and a good agreement of the sulfide results obtained with the AgI resin gel and with other analytical methods. The method was applied on anoxic sediment pore waters in an estuarine and marine system. Simultaneous remobilization of sulfide and trace metals was observed in the marine sediment.

Hydrogel-Forming Microneedles Prepared from ‘‘Super Swelling’’ Polymers Combined with Lyophilised Wafers for Transdermal Drug Delivery

We describe, for the first time, hydrogel-forming microneedle arrays prepared from "super swelling" polymeric compositions. We produced a microneedle formulation with enhanced swelling capabilities from aqueous blends containing 20% w/w Gantrez S-97, 7.5% w/w PEG 10,000 and 3% w/w Na2CO3 and utilised a drug reservoir of a lyophilised wafer-like design. These microneedle-lyophilised wafer compositions were robust and effectively penetrated skin, swelling extensively, but being removed intact. In in vitro delivery experiments across excised neonatal porcine skin, approximately 44 mg of the model high dose small molecule drug ibuprofen sodium was delivered in 24 h, equating to 37% of the loading in the lyophilised reservoir. The super swelling microneedles delivered approximately 1.24 mg of the model protein ovalbumin over 24 h, equivalent to a delivery efficiency of approximately 49%. The integrated microneedle-lyophilised wafer delivery system produced a progressive increase in plasma concentrations of ibuprofen sodium in rats over 6 h, with a maximal concentration of approximately 179 µg/ml achieved in this time. The plasma concentration had fallen to 71±6.7 µg/ml by 24 h. Ovalbumin levels peaked in rat plasma after only 1 hour at 42.36±17.01 ng/ml. Ovalbumin plasma levels then remained almost constant up to 6 h, dropping somewhat at 24 h, when 23.61±4.84 ng/ml was detected. This work represents a significant advancement on conventional microneedle systems, which are presently only suitable for bolus delivery of very potent drugs and vaccines. Once fully developed, such technology may greatly expand the range of drugs that can be delivered transdermally, to the benefit of patients and industry. Accordingly, we are currently progressing towards clinical evaluations with a range of candidate molecules.

Development of a doped titania immobilised thin film multi tubular photoreactor

This paper describes a novel doped titania immobilised thin film multi tubular photoreactor which has been developed for use with liquid, vapour or gas phase media. In designing photocatalytic reactors measuring active surface area of photocatalyst within the unit is one of the critical design parameters. This dictate greatly limits the applicability of any semi-conductor photocatalyst in industrial applications, as a large surface area equates to a powder catalyst. This demonstration of a thin film coating, doped with a rare earth element, novel photoreactor design produces a photocatalytic degradation of a model pollutant (methyl orange) which displayed a comparable degradation achieved with P25 TiO2. The use of lanthanide doping is reported here in the titania sol gel as it is thought to increase the electron hole separation therefore widening the potential useful wavelengths within the electromagnetic spectrum. Increasing doping from 0.5% to 1.0% increased photocatalytic degradation by ∼17% under visible irradiation. A linear relationship has been seen between increasing reactor volume and degradation which would not normally be observed in a typical suspended reactor system. © 2012 Elsevier B.V.

Influence of impurities and catalyst surface characteristics on the oxygen charge transfer reaction in the Pt/YSZ system

Spillover processes (i.e. the migration of ionic species from the support to the catalyst and vice versa) are known to play a very important role in catalysis and electrocatalysis. These spillover processes can be influenced by impurities (pre-existing on the catalyst surface) and by the catalyst morphology that may differ as a result of the differences in catalyst manufacturing processes. This work investigates the influence of impurities present in three commercial platinum (Pt) precursors. The resulting platinum films studied here were supported on yttria-stabilised-zirconia (YSZ). It was found that the three different catalyst films contained a range of impurities (determined by ICP-OES) that appear to affect the oxygen charge transfer reaction as studied by cyclic voltammetry (CV). © 2012 Elsevier B.V.

Hydrogel-forming microneedle arrays made from light-responsive materials for on-demand transdermal drug delivery

We describe, for the first time, stimuli-responsive hydrogel-forming microneedle (MN) arrays that enable delivery of a clinically-relevant model drug (ibuprofen) upon application of light. MN arrays were prepared using a polymer prepared from 2-hydroxyethyl methacrylate (HEMA) and ethylene glycol dimethacrylate (EGDMA) by micromolding. The obtained MN arrays showed good mechanical properties. The system was loaded with up to 5% (w/w) ibuprofen included in a light-responsive 3,5-dimethoxybenzoin conjugate. Raman spectroscopy confirmed the presence of the conjugate inside the polymeric MN matrix. In vitro, this system was able to deliver up to three doses of 50 mg of ibuprofen upon application of an optical trigger over a prolonged period of time (up to 160 hours). This makes the system appealing as a controlled release device for prolonged periods of time. We believe that this technology has potential for use in ?on-demand? delivery of a wide range of drugs in a variety of applications relevant to enhanced patient care.

Automated P.falciparum detection system for post-treatment malaria diagnosis using modified annular ring ratio method

The gametocytes of the malaria parasite Plasmodium falciparum are highly resistant to antimalarial drugs. Its presence in the blood can be detected even after a successful malaria treatment. This paper explains a modified Annular Ring Ratio method which successfully locates and differentiates gametocytes of P. falciparum species in thin blood film images. The method can be used as an efficient tool for gametocyte detection for post-treatment malaria diagnosis. It also identifies the presence of any White Blood Cells (WBCs) in the image, and discards other artifacts and non infected cells. It utilizes the information based on structure, color and geometry of the cells and does not require any segmentation or non-illumination correction techniques that are commonly used for cell detection.

Nanotech thin film photovoltaics

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia de Electrónica e Telecomunicações

Kitchen robot prod. line development. Proj. of Bowden cables cutting and thermal forming machine.

This document presents particular description of work done during student’s internship in PR Metal company realized as ERASMUS PROJECT at ISEP. All information including company’s description and its structure, overview of the problems and analyzed cases, all stages of projects from concept to conclusion can be found here. Description of work done during the internship is divided here into two pieces. First part concerns one activities of the company which is robotic chefs (kitchen robot) production line. Work, that was done for development of this line involved several tasks, among them: creating a single-worker montage station for screwing robots housing’s parts, improve security system for laser welding chamber, what particularly consists in designing automatically closing door system with special surface, that protects against destructive action of laser beam, test station for examination of durability of heating connectors, solving problem with rotors vibrations. Second part tells about main task, realized in second half of internship and stands a complete description of machine development and design. The machine is a part of car handle latch cable production line and its tasks are: cutting cable to required length and hot-forming plastic cover for further assembly needs.

Investigations into the determinations of hydride-forming elements

Analytical methods for the determination of trace amounts of germanium, tin and arsenic were established using hydride generation coupled with direct current plasma atomic emission spectrometry. A continuous gas flowing batch system for the hydride generation was investigated and was applied to the determination of germanium(Ge), tin(Sn), antimony(Sb) and lead(Pb) (Preliminary results suggest that it is also applicable to arsenic)As) ). With this system, the reproducibility of signals was improved and the determination was speeded up, compared with the conventional batch type hydride generation system. Each determination was complete within one minute. Interferences from a number of transition metal ions, especially from Pd(II), Pt(IV), Ni(II), Cu(II), Co(II), and Fe(II, III), have proven to be very serious under normal conditions, in the determination of germanium, tin, and arsenic. These interference effects were eliminated or significantly reduced in the presence of L-cystine or L-cysteine. Thus, a 10-1000 fold excess of Ni(II), Cu(II), Co(II), Fe(II), Pt(IV), Pd(II), etc. can be tolerated without interference, In the presence of L-cystine or L-cysteine, compared with absence of interference reducing agent. The methods for the determination of Ge, Sn, and As were examined by the analyses of standard reference materials. Interference effects from the sample matrix, for example, in transition metal-rich samples, copper, iron and steel, were eliminated by L-cystine (for As and Sn) and by LI cysteine (for Ge). The analysis of a number of standard reference materials gave excellent results of As and Sn contents in agreement with the certified values, showing there was no systematic interference. The detection limits for both germanium and tin were 20 pg ml- I . Preliminary studies were carried out for the determination of antimony and lead. Antimony was found to react with NaBH4, remaInIng from the previous determinations, giving an analytical signal. A reversed injection manner, i.e., injection of the NaBH4 solution prior to the analyte solution was used to avoid uncertainty caused by residual NaBH4 present and to ensure that an excess of NaB H4 was available. A solution of 0.4% L-cysteine was found to reduce the interference from selected transition metal ions, Co(II), Cu(II), Ni(II) and Pt(IV). Hydrochloric acid - hydrogen peroxide, nitric acid - ammonium persulphate, and potassium dichromate malic acid reaction systems for lead hydride generation were compared. The potassium dichromate - malic acid reaction medium proved to be the best with respect to reproducibility and minimal interference. Cu(II), Ni(II), and Fe(II) caused strong interference In lead determinations, which was not reduced by L-cysteine or Lcystine. Sodium citrate, ascorbic acid, dithizone, thiosemicarbazide and penicillamine reduced interferences to some extent. Further interference reduction studies were carried out uSIng a number of amino acids, glycine, alanine, valine, leucine and histidine, as possible interference reducing agents in the determination of germanium. From glycine, alanine, valine to leucine, the interference reduction effect in germanium determinations decreased. Histidine II was found to be very promising In the reduction of interference. In fact, histidine proved more efficient than L-cystine and L-cysteine In the reduction of interference from Ni(II) in the determination of germanium. Signal enhancement by easily ionized elements (EIEs), usually regarded as an interference effect in analysis by DCP-AES, was studied and successfully applied to advantage in improving the sensitivity and detection limit in the determination of As, Ge, Sn, Sb, and Pb. The effect of alkali and alkaline-earth elements on the determination of the above five hydride forming elements was studied. With the appropriate EIE, a signal enhancement of 40-115% was achieved. Linear calibration and good reproducibility were also obtained in the presence of EIEs. III

The versatility and utilization of phosphorus based compounds in classic carbon-carbon bond forming and esterification reactions

The phosphonium salt room temperature ionic liquid tetradecyltrihexylphosphonium chloride (THPC) has been employed as an efficient reusable media for the palladium catalyzed Suzuki cross-coupling reaction of aryl halides, including aryl chlorides, under mild conditions. The cross-coupling reactions were found to proceed in THPC containing small amounts ofwater and toluene (single phase) using potassium phosphate and 1% Pd2(dba)3'CHCI3. Variously substituted iodobenzenes, including electron rich derivatives, reacted efficiently in THPC with a variety of arylboronic acids and were all complete within 1 hour at 50°C. The corresponding aryl bromides also reacted under these conditions with the addition of a catalytic amount of triphenylphosphine that allowed for complete conversion and high isolated yields. The reactions involving aryl chlorides were considerably slower, although the addition of triphenylphosphine and heating at 70°C allowed high conversion of electron deficient derivatives. Addition of water and hexane to the reaction products results in a triphasic system, from which the catalyst was then recycled by removing the top (hexanes) and bottom (aqueous) layers and adding the reagents to the ionic liquid which was heated again at 50°C; resulting in complete turnover of iodobenzene. Repetition of this procedure gave the biphenyl product in 82-97% yield (repeated five times) for both the initial and recycled reaction sequences. IL ESTERIFICATIONREACTION A new class oftrialkylphosphorane has been prepared through reaction of a trialkylphosphine with 2-chlorodimethylmalonate in the presence oftriethylamine. These new reagents promote the condensation reaction of carboxylic acids with alcohols to provide esters along with trialkylphosphine oxide and dimethylmalonate. The condensation reaction of chiral secondary alcohols can be controlled to give either high levels of inversion or retention through a subtle interplay involving basicity of the reaction media, solvent, and tuning the electronic and steric nature of the carboxylic acid and stenc nature of the phosphorane employed. A coherent mechanism is postulated to explain these observations involving reaction via an initial acyloxyphosphonium ion.

Le film de famille, héritier de la tradition orale

Ce mémoire veut démontrer que le film de famille ainsi que ses conditions de réception empruntent, et ce de façon instinctive, au fonctionnement communicatif et rassembleur de la tradition orale. La parole, l’oralité, devrait être considérée comme le catalyseur de la mise en commun du souvenir que le film de famille suscite. Elle est le véhicule de l’interprétation du message du film de famille en tant que discours familial. D’ailleurs, si l’on compare l’influence des deux registres sensoriels présents dans le médium cinématographique (le visuel et l’oralité), le film de famille reprend davantage les modalités de l’oralité. Conséquemment, son contenu, sa forme et sa finalité correspondent à la définition d’un cinéma de l’oralité, un cinéma de la parole défini par Germain Lacasse. En raison d’une absence de travaux portant spécifiquement sur le sujet, l’objectif de cette recherche est de rapprocher et de définir davantage ces liens qui se sont tissés entre la tradition orale et le film de famille. Dans ce dessein, l’approche théorique développée est basée sur les théories de la tradition orale, sur la théorie de la mémoire collective de Maurice Halbwach et sur les rapports entre le cinéma et l’oralité. Ainsi, les aspects suivants sont abordés : Le rôle de l’oralité dans la constitution de la mémoire familiale, l’apport de l’oralité dans les médias stimulateurs de mémoire familiale et finalement, la forme et le contenu du film de famille en tant qu’aspects distinctifs du cinéma oral. Quatre extraits de films de famille québécois des années 20 à aujourd’hui y sont également analysés.

Criminal justice system in pondicherry - a comparative critical study of the past and the presen

The Union Territory of Pondicherry prior to its merger with the Indian Union was a French Colony. The erstwhile territory of Pondicherry along with its hamlets, namely, KARAIKKAL, MAHE and YANAM was administered by the French Regime. Before it was established by French in 1 6 74 A.D. it was part of Vijayanagara Empire. Prior to this, Pondicherry was a part of the Kingdom of Chola and Pallava Kings. During French Regime, the laws which were in force in France in relation to administration of civil and criminal justice were extended to the erstwhile Territory of Pondicherry. Thus while Pondicherry stood influenced by the Inquisitorial system since the beginning of the 18th century, the neighboring states forming part of the Indian Union since Independence came under the Influence of the British system, viz. accusatorial system. The territory of Pondicherry, for administrative reasons, came to be merged with the Indian Union in the early 60's. Following the merger, the Indian administration sought to extent its own laws from time to time replacing erstwhile French Laws, however, subject to certain savings. Thus the transitional period witnessed consequential changes in the administration of the territory, including the sphere of judicial system. Since I 963, the Union Territory of Pondicherry was brought under the spell of the Indian Legal System The people in Pondicherry ' thus have had the benefit of experiencing both the svstems. Their experiences will be of much help to those who undertake comparative studies in law. The plus and minus points of the respective systems help one to develop a detachment that helps independent evaluation of the svstents. The result of these studies could be relevant in revitalising our criminal systems.The present system is evaluated in the light of the past system. New dimensions are added by way' of an empirical study also.