Development, implementation and initial feasibility testing of the MediEmo mobile application to provide support during medically assisted reproduction.

STUDY QUESTION: Is it possible to develop a patient smartphone application for medically assisted reproduction (MAR) that is acceptable to patients and fertility staff? SUMMARY ANSWER: Staff and patients responded positively to the MediEmo smartphone application, perceiving it to be acceptable and feasible to implement in a busy clinic. WHAT IS KNOWN ALREADY: Digital tools are increasingly popular to provide practical, administrative and psychological support alongside medical treatments. Apps and other digital tools have been developed for use alongside MAR but there is very limited research on the development or acceptability and feasibility of these tools. STUDY DESIGN, SIZE, DURATION: Mixed methods research. This article outlines the development phase of the MediEmo smartphone app, which was guided by the Medical Research Council development framework for complex interventions. The resulting MediEmo app was then implemented into a single centre for MAR in the UK, acceptability evaluated and feasibility explored among 1106 potential participants undertaking IVF cycles. PARTICIPANTS/MATERIALS, SETTING, METHODS: Consultation and data collection took part at a single mid-sized urban fertility clinic. Development of the MediEmo smartphone application took place during 2013 to 2017. Implementation of the MediEmo took place from June 2017 to September 2020. The MediEmo app comprises three functions (six features) namely medication management (medication timeline, messaging), mood management (emotional tracking, coping support) and functional support (frequently asked questions, symptom checker). Data on age, fertility diagnosis, anti-Müllerian hormone level were collected about the users of the MediEmo in addition to MediEmo usage data and attitudes towards the MediEmo smartphone application. MAIN RESULTS AND THE ROLE OF CHANCE: Informed by the developmental process described, MediEmo is an app combining patient medication diary management and ease of integration into clinic systems with emotional support, emotional tracking and data capture. This study demonstrates acceptability and feasibility of MediEmo, with good uptake (79.8%), mood data sensitivity and reliability and positive feedback. LIMITATIONS, REASONS FOR CAUTION: Single centre, small number of users in questionnaire studies. WIDER IMPLICATIONS OF THE FINDINGS: The findings suggest smartphone apps can contribute to fertility care and that patient engagement is high. Evaluation of any apps introduced into clinical pathways should be encouraged to promote development of the most useful digital tools for fertility patients. STUDY FUNDING/COMPETING INTEREST(S): This research did not receive any specific grant from any funding agency in the public, commercial or not-for-profit sector. Outside of the submitted work, J.B. reports personal speaker fees from Merck KGaA, Darmstadt, Germany, Merck AB an affiliate of Merck KGaA, Darmstadt Germany, Theramex, MedThink China, Ferring Pharmaceuticals A/S, grant from Merck Serono Ltd, outside the submitted work and that she is co-developer of Fertility Quality of Life (FertiQoL) and MediEmo app; N.M and C.Y are minority shareholders and J.B.'s University (Cardiff University) owns one third of shares. None of the shareholders benefitted financially from MediEmo. I.R., C.H. and K.Y.B.N. declare no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.

The association between government healthcare spending and maternal mortality in the European Union, 1981-2010: a retrospective study.

OBJECTIVE: To determine the association between reductions in government healthcare spending (GHS) on maternal mortality in 24 countries in the European Union (EU) over a 30-year period, 1981-2010. DESIGN: Retrospective study. SETTING AND POPULATION: Twenty-four EU countries (a total population of 419 million as of 2010). METHODS: We used multivariate regression analysis, controlling for country-specific differences in healthcare, infrastructure, population size and demographic structure. GHS was measured as a percentage of gross domestic product. Five-year lag-time analyses were performed to estimate longer standing effects. MAIN OUTCOME MEASURES: Maternal mortality rates. RESULTS: An annual 1% decrease in GHS is associated with significant rises in maternal mortality rates [regression coefficient [R] 0.0177, P = 0.0021, 95% confidence interval [95% CI] 0.0065-0.0289]. For every annual 1% decrease in GHS, we estimate 89 excess maternal deaths in the EU, a 10.6% annual increase in maternal mortality. The impact on maternal mortality was sustained for up to 1 year (R 0.0150, P = 0.0034, 95% CI 0.0050-0.0250). The associations remained significant after accounting for economic, infrastructure and hospital resource controls, in addition to out-of-pocket expenditure, private health spending and total fertility rate. However, accounting for births attended by skilled staff removed the significance of these effects. CONCLUSIONS: Reductions in GHS were significantly associated with increased maternal mortality rates, which may occur through changes in the provision of skilled health professionals attending births. Examples of reduced GHS such as the implementation of austerity measures and budgetary reductions are likely to worsen maternal mortality in the EU.

Immunogenicity and Safety of the AS04-adjuvanted Human Papillomavirus-16/18 Cervical Cancer Vaccine in Malaysian Women Aged 18-35 years: A Randomized Controlled Trial.

INTRODUCTION: Cervical cancer is the third most common cancer in women worldwide. The HPV-16/18 AS04- adjuvanted vaccine (Cervarix©) has previously been shown to be highly immunogenic with a clinically acceptable safety profile. This phase IIIb, double-blind, randomized (1:1) and placebo controlled trial (NCT00345878) was designed to evaluate the vaccine immunogenicity against HPV-16 and HPV-18 as well as its safety and reactogenicity in Malaysian women. METHODS: Healthy women aged 18-35 years received intramuscularly three doses of either the vaccine (HPV group) or aluminium hydroxide (ALU group) at 0, 1, and 6 months. Antibody titers were measured by an enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 271 eligible subjects were enrolled and 266 subjects completed the study. Initially seronegative subjects in the HPV group showed 100% seroconversion one month post-dose-3 for anti HPV-16 and anti-HPV-18 antibodies with geometric mean titers of 11107.5 (95% CI: 9727.3-12683.4) EL.U/mL and 4273.5 (95% CI: 3771.8-4841.9) EL.U/mL, respectively. Over 96% of subjects in both groups received all three vaccine doses. Solicited local (pain) and general symptoms (myalgia, fatigue, arthralgia and headache) were commonly reported in both HPV and ALU groups. Eight serious adverse events were reported throughout the study (five in the HPV group; three in the ALU group), all considered by investigators to be unrelated to vaccination. CONCLUSION: The HPV-16/18 AS04-adjuvanted vaccine was immunogenic and generally well tolerated in Malaysian women aged 18-35 years.

Evolution of beam distribution in crossing a Walkinshaw resonance.

The third-integer coupling resonance at ν(x)-2ν(z)=ℓ, known as the Walkinshaw resonance, is important in high-power accelerators. We find that, when the betatron tunes ramp through a Walkinshaw resonance the fractional emittance growth (FEG) is a universal function of the effective resonance strength: G(1,-2,ℓ)√[ε(xi)]|Δ(ν(x)-2ν(z))/Δn|(-1/2), where G(1,-2,ℓ) is the resonance strength; ε(xi) and ε(zi) are the initial horizontal and vertical emittances, respectively; and |Δ(ν(x)-2ν(z))/Δn| is the resonance crossing rate per revolution. At large effective resonance strengths, the FEG reaches an asymptotic maximum value (FEG)(max)~2ε(xi)/ε(zi) for ε(xi)>>1/2ε(zi) or ε(zi)/(2ε(xi)) for ε(xi)<<1/2ε(zi). There is little emittance exchange at ε(xi)=1/2ε(z), which can be used to minimize emittance growth in crossing a Walkinshaw resonance.

Enterovirus 71 encephalomyelitis and Japanese encephalitis can be distinguished by topographic distribution of inflammation and specific intraneuronal detection of viral antigen and RNA.

AIMS: To investigate if two important epidemic viral encephalitis in children, Enterovirus 71 (EV71) encephalomyelitis and Japanese encephalitis (JE) whose clinical and pathological features may be nonspecific and overlapping, could be distinguished. METHODS: Tissue sections from the central nervous system of infected cases were examined by light microscopy, immunohistochemistry and in situ hybridization. RESULTS: All 13 cases of EV71 encephalomyelitis collected from Asia and France invariably showed stereotyped distribution of inflammation in the spinal cord, brainstem, hypothalamus, cerebellar dentate nucleus and, to a lesser extent, cerebral cortex and meninges. Anterior pons, corpus striatum, thalamus, temporal lobe, hippocampus and cerebellar cortex were always uninflamed. In contrast, the eight JE cases studied showed inflammation involving most neuronal areas of the central nervous system, including the areas that were uninflamed in EV71 encephalomyelitis. Lesions in both infections were nonspecific, consisting of perivascular and parenchymal infiltration by inflammatory cells, oedematous/necrolytic areas, microglial nodules and neuronophagia. Viral inclusions were absent. CONCLUSIONS: Immunohistochemistry and in situ hybridization assays were useful to identify the causative virus, localizing viral antigens and RNA, respectively, almost exclusively to neurones. The stereotyped distribution of inflammatory lesions in EV71 encephalomyelitis appears to be very useful to help distinguish it from JE.

Continuous microalgae cultivation in a photobioreactor.

New biomass sources for alternative fuels has become a subject of increasing importance as the nation strives to resolve the economic and strategic impacts of limited fossil fuel resources on our national security, environment, and global climate. Algae are among the most promising non-food-crop-based biomass feedstocks. However, there are currently no commercially viable microalgae-based production systems for biofuel production that have been developed, as limitations include less-than optimal oil content, growth rates, and cultivation techniques. While batch studies are critical for determining basic growth phases and characteristics of the algal species, steady-state studies are necessary to better understand and measure the specific growth parameters. This study evaluated the effects of dilution rate on microalgal biomass productivity, lipid content, and fatty acid profile under steady-state conditions with continuous illumination and carbon dioxide supplemention for two types of algae. Continuous cultures were conducted for more that 3 months. Our results show that the productivity of Chlorella minutissima varied from 39 to 137 mg/L/day (dry mass) when the dilution rate varied from 0.08 to 0.64 day(-1). The biomass productivity of C. minutissima reached a maximum value (137 mg/L/day) at a dilution rate of 0.33 day(-1), while the productivity of Dunaliella tertiolecta varied from 46 to 91 mg/L/day at a dilution rate of 0.17 to 0.74 day(-1). The biomass productivity of D. tertiolecta reached a maximum value of 91 mg/L/day at a dilution rate of 0.42 day(-1). Moreover, the lipid content had no significant change with various dilution rates.

Culture of microalgae Chlorella minutissima for biodiesel feedstock production.

Microalgae are among the most promising of non-food based biomass fuel feedstock alternatives. Algal biofuels production is challenged by limited oil content, growth rate, and economical cultivation. To develop the optimum cultivation conditions for increasing biofuels feedstock production, the effect of light source, light intensity, photoperiod, and nitrogen starvation on the growth rate, cell density, and lipid content of Chlorella minutissima were studied. The fatty acid content and composition of Chlorella minutissima were also investigated under the above conditions. Fluorescent lights were more effective than red or white light-emitting diodes for algal growth. Increasing light intensity resulted in more rapid algal growth, while increasing the period of light also significantly increased biomass productivity. Our results showed that the lipid and triacylglycerol content were increased under N starvation conditions. Thus, a two-phase strategy with an initial nutrient-sufficient reactor followed by a nutrient deprivation strategy could likely balance the desire for rapid and high biomass generation (124 mg/L) with a high oil content (50%) of Chlorella minutissima to maximize the total amount of oil produced for biodiesel production. Moreover, methyl palmitate (C16:0), methyl oleate (C18:1), methyl linoleate (C18:2), and methyl linolenate (C18:3) are the major components of Chlorella minutissima derived FAME, and choice of light source, intensity, and N starvation impacted the FAME composition of Chlorella minutissima. The optimized cultivation conditions resulted in higher growth rate, cell density, and oil content, making Chlorella minutissima a potentially suitable organism for biodiesel feedstock production.

Solubilization of vorinostat by cyclodextrins.

BACKGROUND: Vorinostat (suberoylanilide hydroxamic acid) is the first histone deacetylase inhibitor approved by US FDA for use in oncology. However, as a hydrophobic acid, its limited aqueous solubility poses a problem for parenteral delivery. Such limited solubility may also affect its oral bioavailability. OBJECTIVE: The aim of this study was to evaluate whether cyclodextrins (CDs), common excipients used in pharmaceutical industry, could increase the aqueous solubility of vorinostat. METHODS: The actual aqueous solubility of vorinostat was investigated by phase-solubility method. Molecular simulation was employed to predict the interaction energy and preferred orientation of vorinostat in CD cavities. RESULTS: Phase-solubility studies indicated that the solubility of vorinostat (7·24×10(-1) mm) was substantially increased when complexed with various CDs, in the following order: randomly methylated-ß-cyclodextrin (RM-ß-CD)>hydroxypropyl-ß-cyclodextrin (HP-ß-CD)>α-cyclodextrin>hydroxypropyl-α-cyclodextrin>Hydroxypropyl-γ-cyclodextrin>γ-cyclodextrin. RM-ß-CD 300 mm increased vorinostat solubility to 70·8 mm, almost two orders of magnitude higher than the baseline solubility. Such findings were in good agreement with the results obtained from molecular simulation. CONCLUSION: CDs, particularly RM-ß-CD and HP-ß-CD, increased vorinostat's solubility. Future studies could be focused on the application of HP-ß-CD in parenteral delivery of vorinostat or using RM-ß-CD as an oral absorption enhancer. Molecular simulation appeared to be a useful tool for the selection of appropriate CD as excipient for drug delivery.

Ten-year review of epidemiology, clinical features, and treatment outcome of achalasia in a regional hospital in Hong Kong.

OBJECTIVE: To describe the epidemiology, clinical features, and treatment outcome of achalasia in Chinese patients. DESIGN: Retrospective study. SETTING: Major regional hospital, Hong Kong. PATIENTS: Clinical records of patients with the diagnosis of achalasia from July 1997 to June 2007 were reviewed. RESULTS: Thirty-two patients were diagnosed with achalasia during the study period. The mean age at diagnosis was 50 years (standard deviation, 20 years). The female-to-male ratio was 1.3:1. The main presenting symptoms were dysphagia (78%) and vomiting (50%). Nine laparoscopic and two open Heller's operations had been performed and 16 patients had undergone endoscopic dilatations. Four patients had botulinum toxin injection and four were taking calcium channel blocker (nifedipine) medications. Botulinum toxin injection and medical therapy had poor short- and long-term responses. Laparoscopic myotomy and pneumatic dilatation had comparable good short- and long-term responses. CONCLUSION: Achalasia affected all age-groups but there was a peak at middle age. Pneumatic dilatation and Heller's myotomy (open or laparoscopic approach) appeared able to maintain longer symptom responses than medical therapy and botulinum toxin injection.

L-Dopa-induced release of cerebral monoamines.

L-Dopa markedly increased the efflux of tritiated dopamine and tritiated serotonin from rat brain slices. This action appeared contingent on the decarboxylation of L-dopa to dopamine, since it could be blocked by an inhibitor of L-amino acid decarboxylase. Selective destruction of catecholamine-containing nerve terminals by 6-hydroxydopamine significantly decreased the uptake and release of tritiated dopamine but not that of tritiated serotonin. These observations support the hypothesis that a portion of exogenously administered L-dopa may enter central serotonin terminals and undergo decarboxylation to the amine with resultant displacement of the endogenous indoleamine from vesicular stores.

Dopamine: stimulation-induced release from central neurons.

Dopamine, synthesized in rat brain slices from labeled L-tyrosine or L-dopa, can be released by electrical stimulation of a type known to induce neuronal depolarization. Pretreatment of the animals with 6-hydroxydopamine, which destroys central catecholamine-containing nerve terminals, substantially reduced the release of dopamine synthesized from [(14)C]tyrosine or from a low concentration of [(3)H]dopa, whereas the release of dopamine formed from a high concentration of [(3)H]dopa remained essentially unchanged. The observations that at high concentrations L-dopa may enter noncatecholaminergic cells, undergo decarboxylation to dopamine, and subsequently be liberated in response to depolarization suggest that dopamine may act as a substitute central transmitter, possibly in serotonergic neurons. This mechanism may contribute to L-dopa's clinical effects in parkinsonian patients.

Patterned immobilization of antibodies in mechanically induced cracks.

A new approach of chemically immobilizing antibody within a pattern based on thin-film cracking is presented. An adjustable pattern width is achieved with resolutions varied from nano- to microscale by using loading stress on thin-film coated elastomer substrate in both one and two dimensions. By introduction of solution or chemical vapor deposition approaches, antibodies were covalently immobilized in the channels. To demonstrate the bioactivity, specificity, and response rate of antibody patterned structure, scanning electron microscopy was used to enumerating bacteria. The chemically coupled antibody is found to retain its specificity when incubated with different bacteria solutions. Trichloro(1H,1H,2H,2H-perfluoroctyl)silane coating on nonsensing regions exhibits a distinguished bacteria-resistant function that is beneficial for providing a low intrinsic background signal in detection. This technique shows a great potential for applications in the fields of sensing and tissue engineering.

Functionalization of AlN surface and effect of spacer density on Escherichia coli pili-antibody molecular recognition.

The immobilization of antibodies to sensor surfaces is critical in biochemical sensor development. In this study, Jeffamine spacers were employed to tether Escherichia coli K99 pilus antibody to AlN/sapphire surfaces which may allow the antibody to freely reorient and potentially improving the antigen capture efficiency. Spacer density was one of the key parameters to be optimized in studying its effect on the immobilization of antibody. The spacer density was controlled by functionalizing AlN/sapphire surfaces with a mixed (3-glycidyloxypropyl)trimethoxysilane (GPTMS) and trichloro(1H,1H,2H,2H-perfluoroctyl)silane (FAS) self-assembled monolayer (SAM) through a step-wise method. Contact angle measurement and X-ray photoelectron spectroscopy (XPS) were used to characterize the surface coverage of GPTMS and surface chemical composition. Compared to spacer fully covered samples, the capture efficiency was improved by approximately 28% with optimal Jeffamine ED 600 spacer density, which depends on the spacer properties such as the number of monomer units and its size.

Effect of surface modification of siliconeon Staphylococcus epidermidis adhesion and colonization.

Cerebrospinal fluid (CSF) shunts for the treatment of hydrocephalus are generally made of silicone rubber. The growth of bacterial colonies on the silicone surface leads to frequent CSF shunt complications. A systematic study of the effect of the surface modification of silicone on Staphylococcus epidermidis adhesion and colonization was performed for different incubation times by means of colony counting and scanning electron microscopy (SEM). Silicone was modified with different biopolymers and silanes, including heparin, hyaluronan, octadecyltrichlorosilane (OTS), and fluoroalkylsilane (FAS) to provide a stable and biocompatible surface with different surface functional groups and degrees of hydrophobicity. The modified silicone surfaces were studied by using contact angle measurements, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). After 4 and 8 h of incubation, the FAS- and OTS-coated silicone and the hyaluronan coated OTS/silicone surfaces showed significantly reduced bacterial adhesion and colonization compared to blank silicone by both quantification methods. However, the heparin coated OTS/silicone showed significantly increased bacterial adhesion. These results indicate that the nature of the surface functional group and surface roughness determine the extent of bacterial adhesion and colonization. However, the degree of hydrophobicity of the surface did not appear to play a determining role in bacterial adhesion and colonization.

Stability of and inflammatory response to silicon coated with a fluoroalkyl self-assembled monolayer in the central nervous system.

A self-assembled monolayer (SAM) of fluoroalkyl silane (FAS) (CF(3)(CF(2))(5)(CH(2))(2)SiCl(3)) was deposited on the surface of silicon wafers, aiming to enhance its stability and to reduce the inflammatory response in the central nervous system. Following implantation of the FAS SAM coated silicon in rat brains, the FAS SAM coating failed to reduce the inflammatory response of silicon, because it could not prevent the corrosion of the underlying silicon. The corrosion was hindered for the initial 10 days by the FAS SAM coating, but there was nearly no difference when compared to the uncoated silicon when the implantation periods were extended to 28 and 90 days. The FAS SAM coating was completely removed within 28 and 90 days. Meanwhile, on all the extracted uncoated and FAS SAM coated silicon wafers, there were proteinaceous substances deposited on the surfaces, and the amount of the deposits increased with exposure time.

Effect of cast molded rifampicin/silicone on Staphylococcus epidermidis biofilm formation.

Infection is one of the most common catheter-related complications, especially in shunt systems used to treat hydrocephalus. Staphylococcus epidermidis is directly related to biomaterial infections owing to its ability to form a biofilm on implanted materials. In this study, scanning electron microscopy (SEM) and atomic force microscopy (AFM) were employed to investigate the effect of the antibiotic rifampicin on the colonization and growth of S. epidermidis 35984 on the surface of silicone. A cast molding method was used to load rifampicin into the silicone precursor before it was cured. Bacteria with a diameter of 800-1000 nm and height of 200-500 nm were found to be embedded in the biofilm. Compact multilayer biofilm structures were found on silicone surfaces upon incubation for 4 and 24 h. On the other hand, sparser biofilm structures were observed on rifampicin-loaded surfaces after incubation for the same duration. Deformation of bacteria was observed by AFM. Moreover, different bacterial colony structures on the surfaces of silicone and rifampicin-loaded silicone were observed by AFM and SEM.

Immobilization of polysaccharides on a fluorinated silicon surface.

A self-assembled monolayer (SAM) of fluoroalkyl silane (FAS) was deposited on a silicon surface by chemical vapor deposition (CVD) at room temperature under 1.01x10(5)Pa nitrogen. Using this new approach, the quality and reproducibility of the SAM are better than those prepared either in solution or by vapor phase deposition, and the deposition process is simpler. In this modified CVD process, the silane monomers, instead of the oligomeric species, are the primary reactants. Full coverage of the silicon surface by FAS molecules was achieved within 5 min. Heparin and hyaluronan, two naturally occurring biocompatible polysaccharides, were successfully covalently attached on the FAS SAM/Si surface by photo-immobilization. Atomic force microscopy (AFM) revealed the morphologic changes after the immobilization of heparin and hyaluronan, and X-ray photoelectron spectroscopy (XPS) confirmed the change in chemical compositions. Such combination of coatings is expected to enhance the stability and biocompatibility of the base material.

Effect of surface proteins on Staphylococcus epidermidis adhesion and colonization on silicone.

Shunt infections are one of the most serious complications in shunt implant surgery. Previous studies have suggested that cerebrospinal fluid (CSF) proteins could affect bacterial adhesion and subsequent shunt infection. A systematic study using immobilized protein on the surface of silane-modified silicone was conducted to determine how these modifications influenced Staphylococcus epidermidis adhesion and colonization. A comparison was also made with silicone having physically adsorbed protein. A colony-counting adhesion assay and scanning electron microscopy (SEM) were used to provide quantitative analysis of bacterial adhesion and semi-quantitative analysis of bacterial colonization, respectively. In order to determine the appropriate silanization process for effective protein immobilization, the effect of bovine serum albumin (BSA) immobilized on n-3-(trimethoxysilyl)propyl-ethylenediamine (AEAPS)/silicone, aminopropyltriethoxysilane (APTMS)/silicone, 3-(glycidyloxypropyl)trimethoxysilane (GPTMS)/silicone, and octadecyltrichlorosilane (OTS)/silicone on bacterial adhesion was investigated. Upon identifying that OTS is the most effective silane, different types of proteins, including: BSA, human serum albumin (HSA), gamma-globulin, and fibrinogen were immobilized on OTS/silicone by a photo-immobilization method. Immobilized protein on modified silicone surfaces was found to be stable in saline for 30 days, while physically adsorbed protein showed instability within hours as determined by contact angle measurements and X-ray photoelectron spectroscopy (XPS). For HSA/OTS/silicone, BSA/OTS/silicone, gamma-globulin/OTS/silicone, fibrinogen/OTS/silicon, and physically absorbed BSA on silicone, the contact angles were 78.5 degrees, 80.7 degrees, 78.9 degrees, 81.3 degrees, and 96.5 degrees; and the amount of nitrogen content was found to be 4.6%, 5.0%, 5.6%, 7.2%, and 3.2%, respectively. All protein immobilized on OTS/silicone surfaces significantly reduced bacterial adhesion by around 75% compared to untreated silicone, while physically adsorbed BSA on silicone reduced by only 29.4%, as determined by colony-counting adhesion assay. However, there was no significant difference on bacterial adhesion among the different types of proteins immobilized on OTS/silicone. Minimizing bacterial adhesion and colonization can be attributed to the increased concentration of -NH2 group, and stability and more hydrophilic nature of the protein/OTS/silicone surfaces.

Liposome-dependent delivery of pteridine antifolates: a two-compartment growth inhibition assay for evaluating drug leakage and metabolism.

We have developed a two-compartment growth inhibition assay that can provide information about leakage, metabolism and delivery of liposome-dependent drugs under cell culture conditions, and at drug concentrations that are relevant to drug delivery. Two cell lines are grown in separate compartments separated from each other by a 0.1 micron polycarbonate membrane. The membrane allows free drugs to diffuse rapidly from one compartment to another, and does not allow liposomes to diffuse through. Liposomes are added to the first compartment, which contains target cells. The extent of leakage caused by these cells is determined by the growth inhibition of non-target cells in the second compartment. We show that methotrexate and methotrexate-gamma-aspartate leak rapidly and almost completely when encapsulated in phosphatidylglycerol/cholesterol (67:33) liposomes. In contrast, there is only 42% leakage when the drugs are encapsulated in distearoylphosphatidylglycerol/cholesterol (67:33) liposomes. We also demonstrate that the target cells (CV1-P) may partially degrade encapsulated methotrexate-gamma-aspartate to methotrexate. Therefore, methotrexate-gamma-aspartate may be a lysosomally cleaved pro-drug of methotrexate.

In vitro haemocompatibility and stability of two types of heparin-immobilized silicon surfaces.

Heparin was covalently immobilized onto a silicon surface by two different methods, carbodiimide-based immobilization and photo-immobilization. In the former method, a (3-aminopropyl) trimethoxysilane (APTMS) self-assembled monolayer (SAM) or multilayer was first coated onto the silicon surface as the bridging layer, and heparin was then attached to the surface in the presence of water-soluble carbodiimide. In the latter method, an octadecyltrichlorosilane (OTS) SAM was coated on the silicon surface as the bridging layer, and heparin was modified by attaching photosensitive aryl azide groups. Upon UV illumination, the modified heparin was then covalently immobilized onto the surface. The hydrophilicity of the silicon surface changed after each coating step, and heparin aggregates on APTMS SAM and OTS SAM were observed by atomic force microscopy (AFM). In vitro haemocompatibility assays demonstrated that the deposition of APTMS SAM, APTMS multilayer and OTS SAM enhanced the silicon's haemocompatibility, which was further enhanced by the heparin immobilization. There is no evident distinction regarding the haemocompatibility between the heparin-immobilized surfaces by both methods. However, heparin on silicon with APTMS SAM and multilayer as the bridging layers is very unstable when tested in vitro with a saline solution at 37 degrees C, due to the instability of APTMS SAM and multilayer on silicon. Meanwhile, photo-immobilized heparin on silicon with OTS SAM as the bridging layer showed superb stability.