A Drug Delivery System for Administration of Anti–TNF-α Antibody

Purpose To describe the fabrication, evaluation, and preliminary in vivo safety of a new drug delivery system (DDS) for topical anti–TNF-α antibody administration. Methods: A DDS was fabricated using inverse template fabrication of a hydrophobic three-dimensional porous scaffold (100–300 μm in diameter porosity) loaded with 10% polyvinyl alcohol hydrogel carrying 5 mg/ml (weight/volume) of anti–TNF-α antibody. Drug-loaded DDS was sterilized with 25 kGy of gamma irradiation. Long-term in vitro antibody affinity and release was evaluated at room temperature or 37°C using enzyme-linked immunosorbent assay (ELISA) and protein fluorescence. In vivo clinical and histolopathological assessment was performed by subcutaneous implantation in BALB/c mice for 3 months. Results: Gamma irradiation, repeated dry/wet cycles, and storage at room temperature for 1 year or 37°C for 1 month had no deleterious effects on antibody affinity. Anti–TNF-α release was high during the first minutes of aqueous exposure, followed by stabilization and gradual, low-dose, antibody release over the next 30 days. Histopathologic evaluation of explanted DDS showed a fibrous pseudocapsule and a myxoid acute/chronic inflammation without granuloma formation surrounding the implants. Conclusions: Sustained local delivery of anti–TNF-α antibody is feasible using the described DDS, which provides stability of the enclosed antibody for up to 1 year of storage. Preliminary results show good in vivo tolerance following subcutaneous placement for 3 months. The proposed fabrication and sterilization process opens new possibilities for the delivery of biologic agents to the anterior surface of the eye. Translational Relevance The described DDS will facilitate the treatment of ocular surface diseases amenable to biologic therapy

Training sector 20, social, educational and legal services

2005 versionTitre de l'écran-titre (visionné le 12 janvier 2010)Première éd. en 1996Traduction de: Techniques policières : programme d'études techniques 310.A0"Training sector 20, social, educational and legal services

Ablation of Potassium-Chloride Cotransporter Type 3 (Kcc3) in Mouse Causes Multiple Cardiovascular Defects and Isosmotic Polyuria.

Inactivation of Kcc3 in a mixed 129/Sv×C57BL/6 mouse background has been previously found to increase systemic blood pressure (BP) through presumed neurogenic mechanisms. Yet, while this background is generally not considered ideal to investigate the cardiovascular system, KCC3 is also expressed in the arterial wall and proximal nephron. In the current study, the effects of Kcc3 ablation was investigated in a pure rather than mixed C57BL/6J background under regular- and high-salt diets to determine whether they could be mediated through vasculogenic and nephrogenic mechanisms. Aortas were also assessed for reactivity to pharmacological agents while isolated from the influence of sympathetic ganglia. This approach led to the identification of unforeseen abnormalities such as lower pulse pressure, heart rate, aortic reactivity and aortic wall thickness, but higher diastolic BP, left ventricular mass and urinary output in the absence of increased catecholamine levels. Salt loading also led systolic BP to be higher, but to no further changes in hemodynamic parameters. Importantly, aortic vascular smooth muscle cells and cardiomyocytes were both found to express KCC3 abundantly in heterozygous mice. Hence, Kcc3 inactivation in our model caused systemic vascular resistance and ventricular mass to increase while preventing extracellular fluid volume to accumulate. Given that it also affected the physiological properties of aortas in vitro, vasculogenic mechanisms could therefore account for a number of the hemodynamic abnormalities observed

Social, educational and legal services : police technology : technical training program, 310.A0 /

2005 versionLa couv. porte en outre: Reach for your dreamsGlossair

Na⁺ loading protocol.

<p>Mice were subjected to 7 days of regular diet (0.26% NaCl, ~10 mg/d) followed by 5 days of high-salt diet (8% NaCl, ~300 mg/d). Days -3 to -1 served as an acclimation period. BP and HR measurements (23 per day) were obtained between 16:30 and 18:45. Those obtained during acclimation, the first 8 daily hemodynamic measurements and the outliers (as defined in <i>Material and Methods</i>) were excluded from further analyses. Chow and water intake, diuresis, fecal output and biochemical parameters were measured at different time points in Nalgene diuresis cages.</p

Body measurements.

<p><b>A,</b> Growth curves. Values shown are from 57 <i>Kcc3</i>^−/− and 98 <i>Kcc3</i>^+/+ animals. Additional measurements are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154398#pone.0154398.t003" target="_blank">Table 3</a>. <b>B and C,</b> Macroscopic appearance of the abdominal cavity. Animals were 15 weeks old and from the same litter. Gonadal depots (indicated by arrows) are much smaller in <i>Kcc3</i>^−/− mouse compared to wild-type mouse.</p

Oligonucleotides used for qPCR studies.

<p>Oligonucleotides used for qPCR studies.</p

Renal function.

<p>Renal function.</p

Hemodynamic parameters.

<p>Measurements were obtained by tail cuff sphygmomanometry using the BP-2000 Series II system. Data are presented as means of daily differences ± SEM between <i>Kcc3</i>^−/− and <i>Kcc3</i>^+/+ mice among 4 consecutive days and 4 to 6 animals per group. <b>A,</b> Regular-salt diet. <b>B,</b> High-salt diet. The protocol used is illustrated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154398#pone.0154398.g001" target="_blank">Fig 1</a>. * indicates that the data are significantly different statistically from 0 (<i>p</i> < 0.05) based on a Wilcoxon rank-sum test. DBP, diastolic blood pressure; HR, heart rate; MAP, mean arterial pressure; PP, pulse pressure; SBP, systolic blood pressure. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154398#pone.0154398.t002" target="_blank">Table 2</a> is used to show the original data from which differences in parameters were calculated under the regular diet.</p