Vaccination inhibits the human adenoviral transduction in a mouse keratoconjunctivitis model

Adenovirus infections are a major cause of epidemic keratoconjunctivitis (EKC), which can lead to corneal subepithelial infiltrates and multifocal corneal opacity. In the current study, we investigated the use of an E1/E3-deleted adenovirus serotype 5 (Ad5) vector as a vaccine administered intramuscularly (IM) or intranasally (IN) against subsequent challenges with a luciferase-expressing Ad5 (Ad5-Luci) vector via eyedrop. We evaluated the adaptive immune response to Ad5 vector vaccination and confirmed a robust polyfunctional CD8 T cell response in splenic cells. Neutralizing Ad5 antibodies were also measured in the sera of vaccinated mice as well as Ad5 antibody in the eye wash solutions. Upon challenge with Ad5-Luci vector 8 weeks post the primary immunization, transduction was significantly reduced by > 70% in the vaccinated mice, which was slightly better in IM- vs. that in IN-vaccinated animals. Resistance to subsequent challenge was observed 10 months post primary IM vaccination, with sustained reduction up to 60% in the Ad5-Luci vector transduction. Passive immunization of naive mice with antisera from IM to vaccinated mice subsequently challenged with the Ad5-Luci vector resulted in approximately 40% loss in transduction efficiency. Furthermore, the mice that received IM immunization with or without CD8 T cell depletion showed > 40% and 70% reductions, respectively, in Ad8 genomic copies after Ad8 topical challenge. We conclude that Ad-vector vaccination successfully induced an adaptive immune response that prevented subsequent Ad transduction in the cornea and conjunctiva-associated tissues in a mouse model of adenovirus keratoconjunctivitis, and that both cellular and humoral immunity play an important role in preventing Ad transduction.     

Near-infrared fluorescence imaging improves the nodal yield in neck dissection in oral cavity cancer – A randomized study

IntroductionLymph node yield (LNY) in neck dissection has been identified as a prognostic factor in oral cavity cancer. The purpose of this study was to investigate the impact of additional use of optical imaging on LNY in therapeutic ND in oral cancer.MethodsConsecutive patients with oral squamous cell carcinoma with clinical neck metastasis planned for primary tumor resection were randomized to conventional neck dissection or near-infrared fluorescence (NIRF)-guided neck dissection, respectively. In the intervention group, patients were injected with ICG-Nanocoll prior to surgery. Intraoperatively, an optical hand-held camera system was used for lymph node identification. Also, NIRF imaging of the neck specimen was performed, and optical signals were pinned with needle markings to guide the pathological examination. The endpoint of the study was LNY per neck side in levels Ib-III.Results31 patients were included with 18 neck sides in the control group and 18 neck sides in the intervention group for evaluation. During NIRF-guided ND, individual lymph nodes could be identified by a bright fluorescent signal and individual tumor-related drainage patterns could be observed in the neck. The LNY in the intervention group was significantly higher compared to the control group (p = 0.032) with a mean of 24 LN (range: 12–33 LN in levels Ib-III compared to 18 LN (range: 10–36 LN) in the control group, respectively.ConclusionsNIRF-guided ND significantly improved the nodal yield compared to the control group. Intraoperative real-time optical imaging enabled direct visualization of tumor-related drainage patterns within the neck lymphatics.     

Light absorbing properties of indocyanine green (ICG) in solution and after adsorption to the retinal surface — an ex-vivo approach

BACKGROUND: To evaluate potential differences in light absorbing properties and stability of indocyanine green (ICG) adsorbed to the retinal surface and of ICG dissolved in water and balanced salt solution. METHODS: The retina of four human donor eyes was prepared by removing the vitreous from the retinal surface. The inner surface of the specimen was covered with two to three drops of a 0.05% or 0.15% ICG solution respectively. After 1 min, the dye was removed by careful irrigation using BSS plus. The retinal specimens were then investigated by diffuse reflection spectroscopy (UV/VIS/NIR Spectrometer Lambda 900/Perkin Elmer equipped with a PELA-1020 integrating sphere accessory) and their absorption evaluated by the Kubelka-Munk function. To control the sensitivity of the setting, diffuse reflectance spectra of ICG adsorbed to a cellulose membrane and Al(2)O(3) were measured. For comparison, absorption spectra of ICG dissolved in water and BSS plus solution were measured in relation to ICG concentration and time using an UV/VIS/NIR Spectrometer Lambda 900/Perkin Elmer. RESULTS: On the retinal surface, absorption spectra exhibited a steep increase of absorption beginning at 620 nm, with a maximum at 736 nm (0.05%) and a shoulder at 745 (0.15%) and a second maximum at approximately 800 nm for both concentrations. Repeated measurement of the retinal surface 13 days after the ICG exposure revealed no changes in the position of the maxima as compared to the initial measurements. Light absorbing properties of ICG on cellulose or Al(2)O(3) are similar to those seen on the retinal surface with respect to the pattern and location of absorption maxima. In contrast, ICG dissolved in water or BSS plus disclosed variations in absorption characteristics depending on dye concentration, solute and time of measurement. CONCLUSIONS: Absorption characteristics and stability of ICG bound to the retinal surface could be of relevance when investigating potential pathomechanisms of ICG related toxicity, which might be related not only to intraoperative but also to postoperative light exposure of patients after intravitreal use of ICG.     

Bacterial ghosts (BGs)—Advanced antigen and drug delivery system

Bacterial ghosts (BGs) are empty bacterial envelopes of Gram-negative bacteria produced by controlled expression of cloned gene E, forming a lysis tunnel structure within the envelope of the living bacteria. BGs are devoid of cytoplasmic content and possess all bacterial bio-adhesive surface properties in their original state while not posing any infectious threat. BGs are ideally suited as an advanced drug delivery system (ADDS) for toxic substances in tumor therapy. The inner space of BGs can be loaded with either single components or combinations of peptides, drugs or DNA which provides an opportunity to design new types of (polyvalent) drug delivery vehicles. Uptake of BGs loaded with Doxorubicin (Dox) by CaCo2 cells led to effective Dox release from endo-lysosomal compartments and accumulation in the nucleus. Viability and proliferative capacity of the cells were significantly decreased (2–3 orders of magnitude) after internalization of Dox loaded BGs as compared to cells incubated with free Dox. The same effect was observed with leukemia cells. Melanoma cells also revealed a high capability to internalize BGs. These results indicate that BGs are able to target a range of types of cancer. BGs have also been investigated as DNA delivery vectors. Studies show DNA loaded BGs are efficiently phagocytosed and internalized by both professional APCs and tumor cells with up to 82% of cells expressing the plasmid-encoded reporter gene. Our studies with BGs as an ADDS system contribute (i) to optimize drug delivery for the treatment of cancer; (ii) define specific conditions for selection and preparation of BG formulations; (iii) and provide a background for the clinical application of BGs in cancer therapy.     

Therapeutic effect of 5-FC on YCD modified murine P388 leukemia in vivo

Cytosine deaminases (CDs) in bacteria andfungi are found to deaminate prodrug 5 - FC intocytotoxic agent 5 - FU .Yeast CD (YCD) is clonedfrom Saccharomyces cerevisiae.It has been shownpreviously that YCD was more efficient inconversing5 - FC than bacterial CD(b CD) [1-3 ] .As anovel suicide gene therapy system,YCD/ 5 - Fcsystem may be promising in cancer therapy andprevention of graft versus host disease.In thepresent study,we established a P388/ DBA murineleukemia model by infusin…     

原发性肝癌患者术前肝储备功能的预测和术后评价

目的 运用脉动色素浓度法(PDD)测定吲哚氰绿潴留率(ICGR15)及有效肝脏血流量(EHBF)评估原发性肝癌患者术前肝脏的储备功能.方法 对55例原发性肝癌患者术前应用PDD法检测ICGR15和EHBF并根据ICGR15分为3组,并行Child-Pugh评分;根据术后肝功的恢复情况将患者分为肝功能恢复良好(G)、轻度不全(M)和重度不全组(S).分析ICGR15三组中术后肝功不全的发生率以及在不同肝功恢复组中ICGR15、EHBF与Child-Pugh评分比较.结果 术后肝功不全在ICGR15三组中的发生率差异具有统计学意义(P＜0.05);肝功恢复不同组间ICGR15、EHBF同Child-Pugh评分比较具有显著性差异(P＞0.05);在不同的Child-Pugh分级之间,ICGR15及EHBF值差异有统计学意义(P＜0.05).结论 ICGR15、EHBF比传统Child-Pugh评分可以更准确的评估肝储备功能并指导确定手术方案.     

论高校大学生绿色网络环境的营造

介绍了网络文化的迅猛发展对高校大学生的学习方式、生活方式、交往方式产生了巨大冲击,尤其对大学生的身心发展产生了不可忽视的消极影响。指出在高校信息化校园建设中,要加强网络文明的建设,提倡和重视绿色网络,并且从高校大学生的实际出发,努力营造一个健康、文明、有序、安全、具有活力、没有污染的绿色网络环境。