PEGylation of paclitaxel largely improves its safety and anti-tumor efficacy following pulmonary delivery in a mouse model of lung carcinoma

Pulmonary delivery offers an attractive route of administration for chemotherapeutic agents, with the advantages of high drug concentrations locally and low side effects systemically. However, fast clearance mechanisms result in short residence time of small molecule drugs in the lungs. Moreover, the local toxicity induced by antineoplastic drugs is considered a major obstacle for the clinical application of inhaled chemotherapy. In this study, we explored the utility of 6 kDa and 20 kDa polyethylene glycol-paclitaxel (PEG-PTX) conjugates to retain paclitaxel within the lungs, achieve its sustained release locally, and thereby, improve its efficacy and reduce its pulmonary toxicity. The conjugates increased the maximum tolerated dose of paclitaxel by up to 100-fold following intratracheal instillation in healthy mice. PEG-PTX conjugates induced lung inflammation. However, the inflammation was lower than that induced by an equivalent dose of the free drug and it was reversible. Conjugation of paclitaxel to both PEG sizes significantly enhanced its anti-tumor efficacy following intratracheal instillation of a single dose in a Lewis lung carcinoma model in mice. PEG-PTX 20k showed equivalent efficacy as PEG-PTX 6k delivered at a 2.5-fold higher dose, suggesting that the molecular weight of the conjugate plays a role in anti-cancer activity. PEG-PTX 20k conjugate presented a prolonged residency and a sustained paclitaxel release within the lungs. This study showed that PEGylation of paclitaxel offers a potential delivery system for inhalation with improved anti-cancer efficacy, prolonged exposure of lung-resident tumors to the antineoplastic drug and reduced local toxicity

Adjuvantation of Pulmonary-Administered Influenza Vaccine with GPI-0100 Primarily Stimulates Antibody Production and Memory B Cell Proliferation

Adjuvants are key components in vaccines, they help in reducing the required antigen dose but also modulate the phenotype of the induced immune response. We previously showed that GPI-0100, a saponin-derived adjuvant, enhances antigen-specific mucosal and systemic antibody responses to influenza subunit and whole inactivated influenza virus (WIV) vaccine administered via the pulmonary route. However, the impact of the GPI-0100 dose on immune stimulation and the immune mechanisms stimulated by GPI-0100 along with antigen are poorly understood. Therefore, in this study we immunized C57BL/6 mice via the pulmonary route with vaccine consisting of WIV combined with increasing amounts of GPI-0100, formulated as a dry powder. Adjuvantation of WIV enhanced influenza-specific mucosal and systemic immune responses, with intermediate doses of 5 and 7.5 μg GPI-0100 being most effective. The predominant antibody subtype induced by GPI-0100-adjuvanted vaccine was IgG1. Compared to non-adjuvanted vaccine, GPI-0100-adjuvanted WIV vaccine gave rise to higher numbers of antigen-specific IgA- but not IgG-producing B cells in the lungs along with better mucosal and systemic memory B cell responses. The GPI-0100 dose was negatively correlated with the number of influenza-specific IFNγ- and IL17-producing T cells and positively correlated with the number of IL4-producing T cells observed after immunization and challenge. Overall, our results show that adjuvantation of pulmonary-delivered WIV with GPI-0100 mostly affects B cell responses and effectively induces B cell memory

High-Voltage Bias Testing Of Thin-Film Pv Modules

Grid-connected PV systems must withstand high voltage bias in addition to harsh environmental conditions. High leakage currents can lead to electromigration and degradation, and thus become important issues for reliability and safety. This paper describes high voltage bias testing of thin-film PV modules in hot and humid conditions in Florida. Sudden rise in RH caused by sudden drop in temperature during the day resulted in sharp peaks in leakage current. Leakage currents for a typical clear sunny day in summer were approximately double compared to corresponding values in winter. Low-resistance paths created with the passage of time increased the leakage current continuously indicating probable exponential growth in leakage currents. Leakage currents increased proportionately to applied voltage bias, at least up to 600 V, showing potential of the technique as an excellent acceleration test

Modified Bassini′s repair: Our experience in a rural hospital setup

Background: Inguinal hernia is a leading cause of work loss and disability. Recurrence and other complications can occur after hernia repair. The aim of this study wasto evaluate the effectiveness of Modified Bassini′s Herniorraphy in themodern days of surgery. Materials and Methods : This is a retrospective study carried out in the MGM HospitalKamothe from 2005 to 2010. Only unilateral uncomplicated inguinal hernia cases were included.All patients had undergone Modified Bassini′s repair. They were followed for 3 years and the complication and recurrence rates were noted. Result: A total of 254 patients were operated by Modified Bassini′s repair only and 241 patients were followed completely and included in the study. The average age was 52.12 + 17.22 years. The mean operation time was 25 + 5.9 min. The mean hospital stay was 3 + 1.1 days. Post-operative pain was minimal in all patients, and was controlled by simple analgesia. Return to work was after 4 weeks. Hematoma and seroma formation requiring drainage were observed in one and two patients, respectively. Scrotal swelling was observed in two patients, which subsided within 2 weeks. Five patients developed urinary retention. No hydrocele, ischemic orchitis or recurrence was found during the follow-up. Wound infection was noted in one patient, which was treated by dressing and oral antibiotics. Recurrence was noted in two (0.83%) patients in the follow-up period of 3 + 0.44 years. Conclusion: Tissue repairs are still used in economically poor patients who cannot afford mesh, with similar results of prosthetic material repair that are commonly used in modern hospitals

Simplifying influenza vaccination during pandemics:sublingual priming and intramuscular boosting of immune responses with heterologous whole inactivated influenza vaccine

The best approach to control the spread of influenza virus during a pandemic is vaccination. Yet, an appropriate vaccine is not available early in the pandemic since vaccine production is time consuming. For influenza strains with a high pandemic potential like H5N1, stockpiling of vaccines has been considered but is hampered by rapid antigenic drift of the virus. It has, however, been shown that immunization with a given H5N1 strain can prime the immune system for a later booster with a drifted variant. Here, we investigated whether whole inactivated virus (WIV) vaccine can be processed to tablets suitable for sublingual (s.l.) use and whether s.l. vaccine administration can prime the immune system for a later intramuscular (i.m.) boost with a heterologous vaccine. In vitro results demonstrate that freeze-drying and tableting of WIV did not affect the integrity of the viral proteins or the hemagglutinating properties of the viral particles. Immunization experiments revealed that s.l. priming with WIV (prepared from the H5N1 vaccine strain NIBRG-14) 4 weeks prior to i.m. booster immunization with the same virus strongly enhanced hemagglutination-inhibition (HI) titers against NIBRG-14 and the drifted variant NIBRG-23. Moreover, s.l. (and i.m.) immunization with NIBRG-14 also primed for a subsequent heterologous i.m. booster immunization with NIBRG-23 vaccine. In addition to HI serum antibodies, s.l. priming enhanced lung and nose IgA responses, while i.m. priming enhanced lung IgA but not nose IgA levels. Our results identify s.l. vaccination as a user-friendly method to prime for influenza-specific immune responses toward homologous and drifted variants

Development Of Sputtering Systems For Large-Area Deposition Of Cuin1-XGaXSe1-YSY Thin-Film Solar Cells

Optimum magnetic field distribution was obtained empirically by selectively removing nickel-coated soft iron pieces at the rear of the magnetron sputtering sources. In turn, the distribution resulted in a thickness uniformity of ±3% over central 10.2 cm length for molybdenum thin films. A completely modified magnetic array in which the magnetic field at the extremities was boosted by introducing strong magnets at and near the periphery, resulting in thickness uniformity for molybdenum of ±2.24% over 10.2 cm, ±2.4% over 12.7 cm, and ±2.95% over 15.2 cm; and zinc oxide, ±2.46% over 10.2 cm, ±3.84% over 12.7 cm, and ±5.6% over 15.2 cm

Preclinical evaluation of topically-administered PEGylated Fab'lung toxicity

PEGylation is a promising approach to increase the residence time of antibody fragments in the lungs and sustain their therapeutic effects. However, concerns arise as to the potential pulmonary toxicity of antibody fragments conjugated to high molecular weight (HMW)polyethylene glycol (PEG), notably after repeated administrations, and the possibility of PEG accumulation in the lungs. The purpose of this proof-of-concept study is to give insights about the safety of lung administration of a Fab’ anti-IL17A antibody fragment conjugated to two-armed 40kDa PEG (PEG40). The presence of the PEG40 moiety inside alveolar macrophages remained stable for at least 24 hours after intratracheal administration of PEG40-Fab’ to mice. PEG40 was then progressively cleared from alveolar macrophages. Incubation of PEG40 alone with macrophages in vitro did not significantly harm macrophages and did not affect phagocytosis or the production of inflammatory markers. After acute or chronic administration of PEG40-Fab’ to mice, no signs of significant pulmonary toxicity or inflammatory cell accumulation were observed. A vacuolization of alveolar macrophages not associated with any inflammation was noticed when PEG40, PEG40-Fab’, or unPEGylated Fab’ were administered. To conclude this preliminary proof of concept study, acute or repeated pulmonary administrations of PEGylated Fab’ appear safe in rodents

Influenza-specific IgG and hemagluttination inhibition (HAI) titers elicited by unadjuvanted and GPI-0100-adjuvanted mucosal influenza vaccine.

<p>Serum samples from the mice described in the legend to Fig. 1 were collected on day 20 and day 34. (A) Total IgG responses after a single immunization. Average 10log IgG titers ± standard error of the mean (S.E.M.), n  = 6 mice per group. The detection limit of the assay is represented by the dotted line. (B) Total IgG responses after two immunizations. (C) HAI titers after two immunizations. The results are expressed as 2log HAI titers of individual mice. The black line represents the geometric mean virus titer per group. Due to technical reasons only 5 and 4 samples from mice receiving 30 µg GPI-0100 adjuvanted intranasal immunization and unadjuvanted intrapulmonary immunization were available for the HAI assay.</p

Effect of GPI-0100 on lung histology.

<p>On day 0 and on day 20 mice received either HBS buffer (A, C, E, G) or GPI-0100 at the indicated doses (B, D, F, H) via the intranasal (A, B, E, F) or the intrapulmonary (C, D, G, H) route. Lung samples were collected 3 days after the second treatment upon termination. Lung sections were prepared and stained for Ly-6G and Ly-6C (A-D) or CD68 (E-H) to identify neutrophils and macrophages, respectively. Representative pictures of histological analyses of each treatment group are shown. The brown colored cells indicated by an arrow is a neutrophil (N) or a macrophage (M).</p

Effect of GPI-0100 on recruitment of inflammatory cells to the lung.

<p>Quantification of (A) neutrophils and (B) macrophages on micrographs as described in the legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052135#pone-0052135-g006" target="_blank">Figure 6</a>. For each staining section, regions of interest (ROI) were selected using HistoQuest software. The total number of cells within the regions were defined by the nucleus staining. Neutrophils or macrophages are defined as nucleated brown cells. The results are presented as % neutrophils or % macrophages per stained sample from individual mouse.</p