Influence Of Delivery Method On Neuroprotection By Bone Marrow Mononuclear Cell Therapy Following Ventral Root Reimplantation With Fibrin Sealant.

The present work compared the local injection of mononuclear cells to the spinal cord lateral funiculus with the alternative approach of local delivery with fibrin sealant after ventral root avulsion (VRA) and reimplantation. For that, female adult Lewis rats were divided into the following groups: avulsion only, reimplantation with fibrin sealant; root repair with fibrin sealant associated with mononuclear cells; and repair with fibrin sealant and injected mononuclear cells. Cell therapy resulted in greater survival of spinal motoneurons up to four weeks post-surgery, especially when mononuclear cells were added to the fibrin glue. Injection of mononuclear cells to the lateral funiculus yield similar results to the reimplantation alone. Additionally, mononuclear cells added to the fibrin glue increased neurotrophic factor gene transcript levels in the spinal cord ventral horn. Regarding the motor recovery, evaluated by the functional peroneal index, as well as the paw print pressure, cell treated rats performed equally well as compared to reimplanted only animals, and significantly better than the avulsion only subjects. The results herein demonstrate that mononuclear cells therapy is neuroprotective by increasing levels of brain derived neurotrophic factor (BDNF) and glial derived neurotrophic factor (GDNF). Moreover, the use of fibrin sealant mononuclear cells delivery approach gave the best and more long lasting results.

Suprachoroidal electrotransfer: a nonviral gene delivery method to transfect the choroid and the retina without detaching the retina.

Photoreceptors and retinal pigment epithelial cells (RPE) targeting remains challenging in ocular gene therapy. Viral gene transfer, the only method having reached clinical evaluation, still raises safety concerns when administered via subretinal injections. We have developed a novel transfection method in the adult rat, called suprachoroidal electrotransfer (ET), combining the administration of nonviral plasmid DNA into the suprachoroidal space with the application of an electrical field. Optimization of injection, electrical parameters and external electrodes geometry using a reporter plasmid, resulted in a large area of transfected tissues. Not only choroidal cells but also RPE, and potentially photoreceptors, were efficiently transduced for at least a month when using a cytomegalovirus (CMV) promoter. No ocular complications were recorded by angiographic, electroretinographic, and histological analyses, demonstrating that under selected conditions the procedure is devoid of side effects on the retina or the vasculature integrity. Moreover, a significant inhibition of laser induced-choroidal neovascularization (CNV) was achieved 15 days after transfection of a soluble vascular endothelial growth factor receptor-1 (sFlt-1)-encoding plasmid. This is the first nonviral gene transfer technique that is efficient for RPE targeting without inducing retinal detachment. This novel minimally invasive nonviral gene therapy method may open new prospects for human retinal therapies.

EPCM-toimitustapamallin kehittäminen Kvaerner Power Oy:n voimalaitostoimituksiin

Tämän diplomityön päämääränäoli laatia suunnitelma voimalaitoksen toimittamiseksi EPCM-toimitustavalla. Työn keskeisiksi tavoitteiksi asetettiin EPCM-toimitustapaan perustuvan toimitustapakonseptin muodostaminen, toimituksiin liittyvien terminologian selkeyttäminen ja harmonisointi sekä voimalaitoksen suhteellisen kustannusrakenteen laatiminen. Työssä tutkittiin voimalaitostoimituksissa tavanomaisesti käytettyjä sopimustyyppejä, kiinteähintaista sopimusta ja kustannusvoittolisäsopimusta sekä harvinaisemmin käytettyjä kattohinta- ja tavoitekustannussopimusta. Vallitsevat toimitustavat, EPC- ja EPCM-toimitustavat sekä monitoimittajatapa, esiteltiin ja niiden vahvuudet sekä heikkoudet selvitettiin. Projektin ominaisuuksilla, markkinoilla ja tilaajan kyvyllä, asiantuntemuksella sekä resursseilla todettiin olevan vaikutusta toimitustavan valintaan. Lisäksi tehtiin katsaus voimalaitostoimitusten kehitystrendeihin ja projektin riskienhallintaan. Voimalaitoksen laitosjärjestelmäjako kehitettiin ja sitä soveltamalla määritettiin voimalaitoksen suhteellinen kustannusrakenne. Kattilaitoksen osuus voimalaitoksen rakentamiskustannuksista todettiin merkittävimmäksi. Laitosjärjestelmien laitekustannusten havaittiin olevan korkeimmat. EPCM-toimitustapamalli kehitettiin teoriaosan pohjalta ja sen rakenne, organisaatio, sopimustyypit ja -suhteetsekä toimituksen riskien jako kuvattiin. Voimalaitoksen rakentamiskustannuksia vertailtiin eri toimitustavoilla ja EPCM-toimitustapamalli todettiin EPC-toimitustapaa edullisemmaksi.

Highly potent delivery method of gp160 envelope vaccine combining lentivirus-like particles and DNA electrotransfer

Particulate antigen assemblies in the nanometer range and DNA plasmids are particularly interesting for designing vaccines. We hypothesised that a combination of these approaches could result in a new delivery method of gp160 envelope HIV-1 vaccine which could combine the potency of virus-like particles (VLPs) and the simplicity of use of DNA vaccines. Characterisation of lentivirus-like particles (lentiVLPs) by western blot, dynamic light scattering and electron microscopy revealed that their protein pattern, size and structure make them promising candidates for HIV-1 vaccines. Although all particles were similar with regard to size and distribution, they clearly differed in p24 capsid protein content suggesting that Rev may be required for particle maturation and Gag processing. In vivo, lentiVLP pseudotyping with the gp160 envelope or with a combination of gp160 and VSV-G envelopes did not influence the magnitude of the immune response but the combination of lentiVLPs with Alum adjuvant resulted in a more potent response. Interestingly, the strongest immune response was obtained when plasmids encoding lentiVLPs were co-delivered to mice muscles by electrotransfer, suggesting that lentiVLPs were efficiently produced in vivo or the packaging genes mediate an adjuvant effect. DNA electrotransfer of plasmids encoding lentivirus-like particles offers many advantages and appears therefore as a promising delivery method of HIV-1 vaccines. Keywords:VLP, Electroporation, Electrotransfer, HIV vaccine, DNA vaccine

Influence of Delivery Method on Neuroprotection by Bone Marrow Mononuclear Cell Therapy following Ventral Root Reimplantation with Fibrin Sealant

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The relationship between socioeconomic status, course delivery method, and student success at a state college: A single institution analysis

In an effort to improve instruction and better accommodate the needs of students, community colleges are offering courses delivered in a variety of delivery formats that require students to have some level of technology fluency to be successful in the course. This study was conducted to investigate the relationship between student socioeconomic status (SES), course delivery method, and course type on enrollment, final course grades, course completion status, and course passing status at a state college. ^ A dataset for 20,456 students of low and not low SES enrolled in science, technology, engineering, and mathematics (STEM) course types delivered using traditional, online, blended, and web enhanced course delivery formats at Miami Dade College, a large open access 4-year state college located in Miami-Dade County, Florida, was analyzed. A factorial ANOVA using course type, course delivery method, and student SES found no significant differences in final course grades when used to determine if course delivery methods were equally effective for students of low and not low SES taking STEM course types. Additionally, three chi-square goodness-of-fit tests were used to investigate for differences in enrollment, course completion and course passing status by SES, course type, and course delivery method. The findings of the chi-square tests indicated that: (a) there were significant differences in enrollment by SES and course delivery methods for the Engineering/Technology, Math, and overall course types but not for the Natural Science course type and (b) there were no significant differences in course completion status and course passing status by SES and course types overall and SES and course delivery methods overall. However, there were statistically significant but weak relationships between course passing status, SES and the math course type as well as between course passing status, SES, and online and traditional course delivery methods. ^ The mixed findings in the study indicate that strides have been made in closing the theoretical gap in education and technology skills that may exist for students of different SES levels. MDC's course delivery and student support models may assist other institutions address student success in courses that necessitate students having some level of technology fluency. ^

Evaluating in vivo delivery of riboflavin with coulomb-controlled iontophoresis for corneal collagen cross-linking: a pilot study.

PURPOSE: To evaluate the efficacy of coulomb-controlled iontophoresis (CCI) for delivery of riboflavin prior to corneal collagen cross-linking (CXL). METHODS: The eyes of 20 8-week-old Lewis rats, subject to epithelium-ON (epi-ON, n = 20 eyes) or epithelium-OFF (epi-OFF, n = 20 eyes) conditions, were used to evaluate the in vivo delivery of two riboflavin solutions: 0.1% riboflavin-20% dextran T500 solution (riboflavin-dextran) and 0.1% riboflavin 5'-phosphate (riboflavin-phosphate). After systemic intramuscular anesthesia, 0.25 mL of the photosensitizing agent was delivered by either instillation or CCI (2.11 mA/cm(2) for 4 or 10 minutes) into either epithelial condition. The CCI probe on the eye without current served as control. Confocal microscopy of flat-mounted corneas was used to analyze intracorneal penetration and fluorometry was used to quantify riboflavin in the aqueous within 30 minutes of treatment. RESULTS: Instillation and CCI allowed for uniform delivery of riboflavin-dextran throughout the stroma after epithelial debridement. Transepithelial delivery of riboflavin-dextran was not efficacious. Riboflavin-phosphate was successfully delivered in both epithelium conditions. Complete saturation of the cornea was achieved using CCI after removing the epithelium, the epi-ON case allowed for limited diffusion. Increasing the time from 4 to 10 minutes greatly increased the amount of riboflavin detected in the cornea and aqueous humor. CONCLUSIONS: Coulomb-controlled iontophoresis is an effective technique for transepithelial delivery of riboflavin-phosphate into the cornea. This drug delivery method would allow clinicians to significantly shorten the time required for the CXL procedure, with or without epithelial debridement. Whether efficient crosslinking can be achieved through an intact epithelium remains to be demonstrated.

A nipple shield delivery system for oral drug delivery to breastfeeding infants : Microbicide delivery to inactivate HIV

A new drug delivery method for infants is presented which incorporates an active pharmaceutical ingredient (API)-loaded insert into a nipple shield delivery system (NSDS). The API is released directly into milk during breastfeeding. This study investigates the feasibility of using the NSDS to deliver the microbicide sodium dodecyl sulfate (SDS), with the goal of preventing mother-to-child transmission (MTCT) of HIV during breastfeeding in low-resource settings, when there is no safer alternative for the infant but to breastfeed. SDS has been previously shown to effectively inactivate HIV in human milk. An apparatus was developed to simulate milk flow through and drug release from a NSDS. Using this apparatus milk was pulsed through a prototype device containing a non-woven fiber insert impregnated with SDS and the microbicide was rapidly released. The total SDS release from inserts ranged from 70 to 100% of the average 0.07 g load within 50 ml (the volume of a typical breastfeed). Human milk spiked with H9/HIVIIIB cells was also passed through the same set-up. Greater than 99% reduction of cell-associated HIV infectivity was achieved in the first 10 ml of milk. This proof of concept study demonstrates efficient drug delivery to breastfeeding infants is achievable using the NSDS.

Nanotechnological approaches using curcumin and Withania somnifera: neuroprotection and antimicrobial activities

Tese de Doutoramento em Biologia de Plantas

The use of the Nursing Activities Score in clinical settings: an integrative review

ABSTRACT Objective analyze how studies have approached the results obtained from the application of the Nursing Activities Score (NAS) based on Donabedian’s model of healthcare organization and delivery. Method CINAHL and PubMed databases were searched for papers published between 2003 and March 2015. Results 36 articles that met the inclusion criteria were reviewed and double-coded by three independent coders and analyzed based on the three elements of Donabedian’s health care quality framework: structure, process and outcome. The most frequently addressed, but not always tested, variables were those that fell into the structure category. Conclusion variables that fell into the process category were used less frequently. Beside NAS, the most frequently used variables in the outcome category were mortality and length of stay. However, no study used a quality framework for healthcare or NAS to evaluate costs, and it is recommended that further research should explore this approach.

Biosafety of Recombinant Adeno-associated Virus Vectors.

It is hoped that the use of gene transfer technology to treat both monogenetic and acquired diseases may soon become a common therapy option in medicine. For gene therapy to achieve this objective, any gene delivery method will have to meet several criteria, including ease of manufacturing, efficient gene transfer to target tissue, long-term gene expression to alleviate the disease, and most importantly safety in patients. Viral vectors are an attractive choice for use in gene therapy protocols due to their relative efficiency in gene delivery. Since there is inherent risk in using viruses, investigators in the gene therapy community have devoted extensive efforts toward reengineering viral vectors for enhance safety. Here we review the approaches and technologies that are being evaluated for the use of recombinant vectors based upon adeno-associated virus (AAV) in the treatment of a variety of human diseases. AAV is currently the only known human DNA virus that is non-pathogenic and AAV-based vectors are classified as Risk Group 1 agents for all laboratory and animal studies carried out in the US. Although its apparent safety in natural infection and animals appears well documented, we examine the accumulated knowledge on the biology and vectorology of AAV, lessons learned from gene therapy clinical trials, and how this information is impacting current vector design and manufacturing with an overall emphasis on biosafety.

In vivo reprogramming of circuit connectivity in postmitotic neocortical neurons.

The molecular mechanisms that control how progenitors generate distinct subtypes of neurons, and how undifferentiated neurons acquire their specific identity during corticogenesis, are increasingly understood. However, whether postmitotic neurons can change their identity at late stages of differentiation remains unknown. To study this question, we developed an electrochemical in vivo gene delivery method to rapidly manipulate gene expression specifically in postmitotic neurons. Using this approach, we found that the molecular identity, morphology, physiology and functional input-output connectivity of layer 4 mouse spiny neurons could be specifically reprogrammed during the first postnatal week by ectopic expression of the layer 5B output neuron-specific transcription factor Fezf2. These findings reveal a high degree of plasticity in the identity of postmitotic neocortical neurons and provide a proof of principle for postnatal re-engineering of specific neural microcircuits in vivo.

Mobile learning and video experience in delivering digital content

In recent years, the worldwide distribution of smartphone devices has been growing rapidly. Mobile technologies are evolving fast, a situation which provides new possibilities for mobile learning applications. Along with new delivery methods, this development enables new concepts for learning. This study focuses on the effectiveness and experience of a mobile learning video promoting the key features of a specific device. Through relevant learning theories, mobile technologies and empirical findings, the thesis presents the key elements for a mobile learning video that are essential for effective learning. This study also explores how previous experience with mobile services and knowledge of a mobile handset relate to final learning results. Moreover, this study discusses the optimal delivery mechanisms for a mobile video. The target group for the study consists of twenty employees of a Sanoma Company. The main findings show that the individual experience of learning and the actual learning results may differ and that the design for certain video elements, such as sound and the presentation of technical features, can have an impact on the experience and effectiveness of a mobile learning video. Moreover, a video delivery method based on cloud technologies and HTML5 is suggested to be used in parallel with standalone applications.