Increased potential of a cationic liposome-based delivery system:enhancing stability and sustained immunological activity in pre-clinical development

The combination of dimethyl dioctadecyl ammonium bromide (DDA) and the synthetic cord factor trehalose dibehenate (TDB) with Ag85B-ESAT-6 (H1 fusion protein) has been found to promote strong protective immune responses against Mycobacterium tuberculosis. The development of a vaccine formulation that is able to facilitate the requirements of sterility, stability and generation of a vaccine product with acceptable composition, shelf-life and safety profile may necessitate selected alterations in vaccine formulation. This study describes the implementation of a sterilisation protocol and the use of selected lyoprotective agents in order to fulfil these requirements. Concomitantly, close analysis of any alteration in physico-chemical characteristics and parameters of immunogenicity have been examined for this promising DDA liposome-based tuberculosis vaccine. The study addresses the extensive guidelines on parameters for non-clinical assessment, suitable for liposomal vaccines and other vaccine delivery systems issued by the World Health Organisation (WHO) and the European Medicines Agency (EMEA). Physical and chemical stability was observed following alteration in formulations to include novel cryoprotectants and radiation sterilisation. Immunogenicity was maintained following these alterations and even improved by modification with lysine as the cryoprotective agent for sterilised formulations. Taken together, these results outline the successful alteration to a liposomal vaccine, representing improved formulations by rational modification, whilst maintaining biological activity.

Towards an improved ocular drug delivery system

The ultimate aim of this project was to design new biomaterials which will improve the efficiency of ocular drug delivery systems. Initially, it was necessary to review the information available on the nature of the tear fluid and its relationship with the eye. An extensive survey of the relevant literature was made. There is a common belief in the literature that the ocular glycoprotein, mucin, plays an important role in tear film stability, and furthermore, that it exists as an adherent layer covering the corneal surface. If this belief is true, the muco-corneal interaction provides the ideal basis for the development of sustained release drug delivery. Preliminary investigations were made to assess the ability of mucin to adhere to polymer surfaces. The intention was to develop a synthetic model which would mimic the supposed corneal/mucin interaction. Analytical procedures included the use of microscopy (phase contrast and fluorescence), fluorophotometry, and mucin-staining dyes. Additionally, the physical properties of tears and tear models were assessed under conditions mimicking those of the preocular environment, using rheological and tensiometric techniques. The wetting abilities of these tear models and opthalmic formulations were also investigated. Tissue culture techniques were employed to enable the surface properties of the corneal surface to be studied by means of cultured corneal cells. The results of these investigations enabled the calculation of interfacial and surface characteristics of tears, tear models, and the corneal surface. Over all, this work cast doubt on the accepted relationship of mucin with the cornea. A corneal surface model was designed, on the basis of the information obtained during this project, which would possess similar surface chemical properties (i.e. would be biomimetic) to the more complex original. This model, together with the information gained on the properties of tears and solutions intended for ocular instillation, could be valuable in the design of drug formulations with enhanced ocular retention times. Furthermore, the model itself may form the basis for the design of an effective drug-carrier.

Subunit vaccines distearoylphosphatidylcholine-based liposomes entrapping antigen offer a neutral alternative to dimethyldioctadecylammonium-based cationic liposomes as an adjuvant delivery system

The adjuvanticity of liposomes can be directed through formulation to develop a safe yet potent vaccine candidate. With the addition of the cationic lipid dimethyldioctadecylammonium bromide (DDA) to stable neutral distearoylphosphatidylcholine (DSPC):cholesterol (Chol) liposomes, vesicle size reduces while protein entrapment increases. The addition of the immunomodulator, trehalose 6,6-dibehenate (TDB) to either the neutral or cationic liposomes did not affect the physiochemical characteristics of these liposome vesicles. However, the protective immune response, as indicated by the amount of IFN-? production, increases considerably when TDB is present. High levels of IFN-? were observed for cationic liposomes; however, there was a marked reduction in IFN-? release over time. Conversely, for neutral liposomes containing TDB, although the initial amount of IFN-? was slightly lower than the cationic equivalent, the overall protective immune responses of these neutral liposomes were effectively maintained over time, generating good levels of protection. To that end, although the addition of DSPC and Chol reduced the protective immunity of DDA:TDB liposomes, relatively high protection was observed for the neutral counterpart, DSPC:Chol:TDB, which may offer an effective neutral alternative to the DDA:TDB cationic system, especially for the delivery of either zwitterionic (neutral) or cationic molecules or antigens.

Polycaprolactone fibres as a potential delivery system for collagen to support bone regeneration

Poly(e-caprolactone) (PCL) is biocompatible, non-immunogenic and non-toxic, and slowly degrades, allowing sufficient time for tissue regeneration. PCL has the potential for application in bone and cartilage repair as it may provide the essential structure required for bone regeneration, however, an ideal scaffold system is still undeveloped. PCL fibres were prepared using the gravity spinning technique, in which collagen was either incorporated into or coated onto the 'as-spun' fibres, in order to develop novel biodegradable polymer fibres which will effectively deliver collagen and support the attachment and proliferation of human osteoblast (HOB) cells for bone regeneration. The physical and mechanical characteristics and cell fibre interactions were analysed. The PCL fibres were found to be highly flexible and inclusion of collagen did not alter the mechanical properties of PCL fibres. Overall, HOB cells were shown to effectively adhere and proliferate on all fibre platforms tested, although proliferation rates were enhanced by surface coating PCL fibres with collagen compared to PCL fibres incorporating collagen and PCL-only fibres. These findings highlight the potential of using gravity spun PCL fibres as a delivery platform for extracellular matrix proteins, such as collagen, in order to enhance cell adherence and proliferation for tissue repair.

Capillary siphons and their application in the fuel delivery system of direct methanol fuel cells

The objective of the work is to develop a fuel delivery system for potable direct methanol fuel cell. Currently, one of the most fundamental limitations of direct methanol fuel cells is that the fuel supplied to the anode of the DMFC must be a very dilute aqueous methanol solution (usually 0.5∼1.5 M). If a DMFC is filled with a dilute aqueous methanol solution, the fuel cell operation time per refuel would be very short, which would considerably diminish the advantage of a DMFC over a conventional battery. To overcome this difficulty, a complex fuel delivery system based on the modern micro system technology was proposed by the author. The proposed fuel delivery system would include micro-pumps, a methanol sensor, and a control unit. The fuel delivery system adds considerable costs to the fuel cell system and consume considerable amount of electricity from the fuel cell, which in turn significantly reduces the net power output of the fuel cell. As a result, the DMFC would have tremendous difficulty to compete with the conventional battery technology in terms of costs and power output. ^ This work presents a novel passive fuel delivery system for direct methanol fuel cells. In this particular system, a methanol fuel and an aqueous methanol solution are stored separately in two containers and a wick is disposed between the two containers in a siphon fashion, with the container of the aqueous methanol solution communicating with the anode of the DMFC. Methanol is siphoned from the methanol container to the aqueous solution container in-situ when the methanol in the aqueous methanol solution is consumed during the operation of the fuel cell. Through a proper selection of the wick and the containers, the methanol concentration near the anode of the DMFC could be maintained within a preferable range. ^

Integrated construction project delivery system in the U.S. public sector: An information modeling framework

Integrated project delivery (IPD) method has recently emerged as an alternative to traditional delivery methods. It has the potential to overcome inefficiencies of traditional delivery methods by enhancing collaboration among project participants. Information and communication technology (ICT) facilitates IPD by effective management, processing and communication of information within and among organizations. While the benefits of IPD, and the role of ICT in realizing them, have been generally acknowledged, the US public construction sector is very slow in adopting IPD. The reasons are - lack of experience and inadequate understanding of IPD in public owner as confirmed by the results of the questionnaire survey conducted under this research study. The public construction sector should be aware of the value of IPD and should know the essentials for effective implementation of IPD principles - especially, they should be cognizant of the opportunities offered by advancements in ICT to realize this.^ In order to address the need an IPD Readiness Assessment Model (IPD-RAM) was developed in this research study. The model was designed with a goal to determine IPD readiness of a public owner organization considering selected IPD principles, and ICT levels, at which project functions were carried out. Subsequent analysis led to identification of possible improvements in ICTs that have the potential to increase IPD readiness scores. Termed as the gap identification, this process was used to formulate improvement strategies. The model had been applied to six Florida International University (FIU) construction projects (case studies). The results showed that the IPD readiness of the organization was considerably low and several project functions can be improved by using higher and/or advanced level ICT tools and methods. Feedbacks from a focus group comprised of FIU officials and an independent group of experts had been received at various stages of this research and had been utilized during development and implementation of the model. Focus group input was also helpful for validation of the model and its results. It was hoped that the model developed would be useful to construction owner organizations in order to assess their IPD readiness and to identify appropriate ICT improvement strategies.^

Integrated Construction Project Delivery System in the U.S. Public Sector: An Information Modeling Framework

Integrated project delivery (IPD) method has recently emerged as an alternative to traditional delivery methods. It has the potential to overcome inefficiencies of traditional delivery methods by enhancing collaboration among project participants. Information and communication technology (ICT) facilitates IPD by effective management, processing and communication of information within and among organizations. While the benefits of IPD, and the role of ICT in realizing them, have been generally acknowledged, the US public construction sector is very slow in adopting IPD. The reasons are - lack of experience and inadequate understanding of IPD in public owner as confirmed by the results of the questionnaire survey conducted under this research study. The public construction sector should be aware of the value of IPD and should know the essentials for effective implementation of IPD principles - especially, they should be cognizant of the opportunities offered by advancements in ICT to realize this. In order to address the need an IPD Readiness Assessment Model (IPD-RAM) was developed in this research study. The model was designed with a goal to determine IPD readiness of a public owner organization considering selected IPD principles, and ICT levels, at which project functions were carried out. Subsequent analysis led to identification of possible improvements in ICTs that have the potential to increase IPD readiness scores. Termed as the gap identification, this process was used to formulate improvement strategies. The model had been applied to six Florida International University (FIU) construction projects (case studies). The results showed that the IPD readiness of the organization was considerably low and several project functions can be improved by using higher and/or advanced level ICT tools and methods. Feedbacks from a focus group comprised of FIU officials and an independent group of experts had been received at various stages of this research and had been utilized during development and implementation of the model. Focus group input was also helpful for validation of the model and its results. It was hoped that the model developed would be useful to construction owner organizations in order to assess their IPD readiness and to identify appropriate ICT improvement strategies.

Improving the Utilization of Front-Line Service Delivery System Personnel

There are two types of work typically performed in services which differ in the degree of control management has over when the work must be done. Serving customers, an activity that can occur only when customers are in the system is, by its nature, uncontrollable work. In contrast, the execution of controllable work does not require the presence of customers, and is work over which management has some degree of temporal control. This paper presents two integer programming models for optimally scheduling controllable work simultaneously with shifts. One model explicitly defines variables for the times at which controllable work may be started, while the other uses implicit modeling to reduce the number of variables. In an initial experiment of 864 test problems, the latter model yielded optimal solutions in approximately 81 percent of the time required by the former model. To evaluate the impact on customer service of having front-line employees perform controllable work, a second experiment was conducted simulating 5,832 service delivery systems. The results show that controllable work offers a useful means of improving labor utilization. Perhaps more important, it was found that having front-line employees perform controllable work did not degrade the desired level of customer service.

Dual stimulation of antigen presenting cells using carbon nanotube-based vaccine delivery system for cancer immunotherapy

Although anti−cancer immuno−based combinatorial therapeutic approaches have shown promising results, efficient tumour eradication demands further intensification of anti−tumour immune response. With the emerging field of nanovaccinology, multi−walled carbon nanotubes (MWNTs) have manifested prominent potentials as tumour antigen nanocarriers. Nevertheless, the utilization of MWNTs in co−delivering antigen along with different types of immunoadjuvants to antigen presenting cells (APCs) has not been investigated yet. We hypothesized that harnessing MWNT for concurrent delivery of cytosine−phosphate−guanine oligodeoxynucleotide (CpG) and anti-CD40 Ig (αCD40), as immunoadjuvants, along with the model antigen ovalbumin (OVA) could potentiate immune response induced against OVA−expressing tumour cells. We initially investigated the effective method to co−deliver OVA and CpG using MWNT to the APC. Covalent conjugation of OVA and CpG prior to loading onto MWNTs markedly augmented the CpG−mediated adjuvanticity, as demonstrated by the significantly increased OVA−specific T cell responses in vitro and in C57BL/6 mice. αCD40 was then included as a second immunoadjuvant to further intensify the immune response. Immune response elicited in vitro and in vivo by OVA, CpG and αCD40 was significantly potentiated by their co−incorporation onto the MWNTs. Furthermore, MWNT remarkably improved the ability of co−loaded OVA, CpG and αCD40 in inhibiting the growth of OVA−expressing B16F10 melanoma cells in subcutaneous or lung pseudo−metastatic tumour models. Therefore, this study suggests that the utilization of MWNTs for the co−delivery of tumour−derived antigen, CpG and αCD40 could be a competent approach for efficient tumours eradication.

Evaluation Of An Extended Duct Air Delivery System For Spaces Conditioned By Rooftop Units

Traditional air delivery to high-bay buildings involves ceiling level supply and return ducts that create an almost-uniform temperature in the space. Problems with this system include potential recirculation of supply air and higher-than-necessary return air temperatures. A new air delivery strategy was investigated that involves changing the height of conventional supply and return ducts to have control over thermal stratification in the space. A full-scale experiment using ten vertical temperature profiles was conducted in a manufacturing facility over one year. The experimental data was utilized to validated CFD and EnergyPlus models. CFD simulation results show that supplying air directly to the occupied zone increases stratification while holding thermal comfort constant during the cooling operation. The building energy simulation identified how return air temperature offset, set point offset, and stratification influence the building’s energy consumption. A utility bill analysis for cooling shows 28.8% HVAC energy savings while the building energy simulation shows 19.3 – 37.4% HVAC energy savings.

Methotrexate nanoparticle delivery system for treatment of inflammatory bowel disease in pediatric patients

Purpose: To evaluate the efficacy and safety of methotrexate (MTX) nanoparticles in pediatric patients with inflammatory bowel disease (IBD). Methods: In this randomized, open-label clinical study, 28 pediatric patients with moderate to severe IBD were randomly assigned to treatment (MTX nanoparticles,15 mg/week) or control (azathioprine, AZA, 2 mg/kg/day) group. Nanoparticles were synthesized by adding calcium chloride to sodium alginate solution containing MTX, and was further treated with poly-L-lysine aqueous solution. The nanoparticles were evaluated for particle size, zeta potential and drug encapsulation efficacy. Erythrocyte sedimentation rate, C-reactive protein, aspartate aminotransferase, alanine transaminase, and disease activity scores were used to assess IBD remission. Results: Nanoparticle size, zeta potential and encapsulation efficacy were 164.4 ± 6.9 nm, -32.6 ± 3.7 mV, and 97.8 ± 4.2 %, respectively. After 12 weeks of therapy, the mean Pediatric Crohn\'s Disease Activity Index (PCDAI) scores for control and treatment groups were 22.3 ± 2.14 and 16.8 ± 1.87, respectively, while mean Pediatric Ulcerative Colitis Activity (PUCAI) Index scores were 24.3 ± 1.47 and 18.7 ± 1.92, respectively. Eight patients in the treatment and five patients in the control group achieved remission. Biochemical parameters varied significantly between the groups. Conclusion: MTX nanoparticles are safe and more effective than standard first-line IBD therapy. However, further studies are required to determine the suitability of the formulation for therapeutic use.

Bioadhesive drug delivery system of diltiazem hydrochloride for improved bioavailability in cardiac therapy

Purpose: To prepare and evaluate bioadhesive buccal films of diltiazem hydrochloride (a L-type calcium channel blocker) for overcoming the limitations of frequent dosing, low bioavailability and gastrointestinal discomfort of oral delivery. Methods: Buccal films were prepared by solvent casting technique using sodium carboxymethylcellulose, polyvinyl pyrrolidone K-30 and polyvinyl alcohol. The films were evaluated for weight, thickness, surface pH, swelling index, in vitro residence time, folding endurance, in vitro release, ex-vivo permeation (across porcine buccal mucosa) and drug content uniformity. Results: The drug content of the formulations was uniform with a range of 18.94 ± 0.066 (F2) to 20.08 ± 0.07 mg per unit film (F1). The films exhibited controlled release ranging from 58.76 ± 1.62 to 91.45 ± 1.02 % over a period > 6 h. The films containing 20 mg diltiazem hydrochloride, polyvinyl alcohol (10 %) and polyvinyl pyrrolidone (1 % w/v) i.e. formulation F5, showed moderate swelling, convenient residence time and promising drug release, and thus can be selected for further development of a buccal film for potential therapeutic uses. Conclusion: The developed formulation is a potential bioadhesive buccal system for delivering diltiazem directly to systemic circulation, circumventing first-pass metabolism, avoiding gastric discomfort and improving bioavailability at a minimal dose.