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Inefficient conjunction search made efficient by concurrent spoken delivery of target identity.

Visual search based on a conjunction of two features typically elicits reaction times that increase linearly as a function of the number of distractors, whereas search based on a single feature is essentially unaffected by set size. These and related findings have often been interpreted as evidence of a serial search stage that follows a parallel search stage. However, a wide range of studies has been showing a form of blending of these two processes. For example, when a spoken instruction identifies the conjunction target concurrently with the visual display, the effect of set size is significantly reduced, suggesting that incremental linguistic processing of the first feature adjective and then the second feature adjective may facilitate something approximating a parallel extraction of objects during search for the target. Here, we extend these results to a variety of experimental designs. First, we replicate the result with a mixed-trials design (ruling out potential strategies associated with the blocked design of the original study). Second, in a mixed-trials experiment, the order of adjective types in the spoken query varies randomly across conditions. In a third experiment, we extend the effect to a triple-conjunction search task. A fourth (control) experiment demonstrates that these effects are not due to an efficient odd-one-out search that ignores the linguistic input. This series of experiments, along with attractor-network simulations of the phenomena, provide further evidence toward understanding linguistically mediated influences in real-time visual search processing.

Attention↗

A DNA delivery system targeting dendritic cells for use in immunization against malaria: a rodent model.

DNA-based vaccination has emerged as a promising method of immunisation since the first demonstration of this technology. Improving the antibody responses is desirable for the protective efficacy and hence broad application of these vaccines. We examined the immunogenicity of a Plasmodium-based DNA vaccine that was targeted to antigen presenting cells by fusion to CTLA4. Fusion proteins comprising the extra-cellular domain of CTLA4, the hinge, CH2 and CH3 domains of human IgG1 and MSP-1 gene fragments were expressed in COS-7 cells. Three of the secreted proteins containing the mouse homologue of CTLA4 were shown to bind differently to the human B7-1 molecule expressed on THP-1 cells. Competition binding assays for two fusion proteins showed that binding was specific. When C57BL/6 mice were immunized with plasmids encoding the fusion proteins, antibodies against two denatured and one non-denatured MSP-1 gene fragments were successfully induced. The usefulness of this strategy in future studies of immunisaton against human malaria is discussed.

Animals↗

Cancer cell-targeted drug delivery utilizing oligopeptide transport activity.

To study the drug delivery to tumor by utilization of an oligopeptide transport activity, we examined the accumulation of dipeptides and the peptide-mimetic anti-cancer drug, bestatin, a substrate of oligopeptide transporter PepT1. Firstly, we established HeLa cells stably expressing human peptide transporter (hPepT1) (HeLa-hPepT1). Secondly, we constructed an experimental model by inoculation of HeLa-hPepT1 cells subcutaneously into Balb/c nu/nu mice to demonstrate the contribution of PepT1 to the tissue-selective drug delivery. The accumulations of a hydrolysis-resistant dipeptide [(3)H]carnosine and bestatin in solid tumors formed by HeLa-hPepT1 or HeLa-pcDNA3, which are transfected with vector DNA (pcDNA3) were measured. After I.V. administration, tissue-to-plasma concentration ratios (K(p)) of both compounds, in HeLa-hPepT1 tumor was significantly greater than that of [(14)C]inulin, a marker for extracellular fluid space, those of dipeptides in muscle, or those in HeLa-pcDNA3 tumor. Furthermore, bestatin exhibited growth inhibition of HeLa-hPepT1 in vitro. In vivo, repeated oral administration of bestatin for 28 days suppressed the growth of HeLa-hPepT1 tumor specifically. When HT-1080 cells, which may naturally express oligopeptide transport activity, were transplanted, K(p) of [(3)H]carnosine was significantly increased in comparison with that in muscle. In addition, oligopeptide transport activities among various human cell lines were examined. These results provide the first demonstration for the selective delivery of oligopeptides to tumors by specific oligopeptide transport activity.

Animals↗

[A novel gene delivery system targeting to epidermal growth factor receptor overexpressing cancer cells].

A polypeptide with 33 amino acid residues was designed and synthesized. Its C terminal was composed of multiple lysine residues, which played as a DNA condensing agent, whereas the N terminal was the receptor binding domain of Epidermal Growth Factor(N32-K48). Through a spontaneous self-assembly process with electrostatic interaction, the synthetic peptide combined with a luciferase expression vector, pEBluc, to form an EGF receptor targeting nucleic acid complex. Significant luciferase activity was detected 48 hours after adding this complex directly to the culture medium of the A431 cells. This synthetic peptide could be used to construct a gene transfer system mediated by the endocytosis via EGF receptor. It promoted a very possibility of the gene therapy for the cancers such as glioma, melanoma and squamous carcinoma which are known of epidermal growth factor receptor overexpression.

Carcinoma↗

Optimization of tumor-targeted gene delivery by engineered attenuated Salmonella typhimurium.

BACKGROUND: Attenuated Salmonella typhimurium has been demonstrated as a potential gene delivery vector. Previous findings induce the necessity to optimize tumor selectivity and bacterial dosing in relation to tumor volume and intratumoral therapeutic gene expression. MATERIALS AND METHODS: Attenuated Salmonella VNP20009 and VNP20047 (expressing cytosine deaminase) were systemically administered to tumor-bearing rats. The bacteria were quantified in tumor and normal organs. Conversion of 5-fluorocytosine to 5-fluorouracil was evaluated using thin layer chromatography. RESULTS: Tumor colonization efficiency was dependent on Salmonella density, administration route and tumor volume. Colonization of normal tissues gradually decreased with time, while intratumoral proliferation of bacteria remained high during the follow-up period. The Optimal Therapeutic Dose (OTD) was found to be 5.10(7) cfu/rat. Intratumoral VNP20047-expressed CDase leading to the conversion of 5-FC to 5-FU was detected in vivo. CONCLUSION: Our results indicate the need to define an OTD, probably for each species, when using genetically engineered Salmonella as a tumor- and species-selective vector in cancer therapy.

Animals↗

Targeted gene delivery to human osteosarcoma cells with magnetic cationic liposomes under a magnetic field.

Gene delivery using cationic liposomes results in relatively low transfection, especially under in vivo conditions. This system, however, can overcome some of the problems associated with viral delivery systems. The present study was carried out in order to improve the transfection efficiency of cationic liposomes by preparing magnetic cationic liposomes (MCL). Small MCL approximately 40 nm in diameter and incorporating one or two magnetite particles were prepared with phosphatidylethanolamine and 3beta-[N-(N', N'-dimethylaminoethane)-carbamoyl] cholesterol. The efficiency of MCL in gene delivery was evaluated by using plasmid DNA containing a luciferase reporter gene and human osteosarcoma Saos-2 cells. Without a magnetic field, maximum luciferase activity was observed when DNA was mixed with MCL at a 1:5 ratio and incubated with cells for 6 h. Under a magnetic field, maximum luciferase activity was achieved by 30-min magnetic induction. This improvement in transfection efficiency by magnetic induction was approximately 3.5-fold. The feasibility of this active transgenic system was further shown by measuring apoptosis rates after transfection of the p53 gene to Saos-2 cells. Apoptosis rates increased to 18.9% from 2.4% by magnetic induction. In conclusion, a gene delivery system including MCL and magnetic induction was found to achieve rapid and enhanced gene delivery in vitro. Such a gene delivery system may be applicable under in vivo conditions, and is expected to offer numerous clinical advantages.

Bone Neoplasms↗

Modern Approaches to Cancer Therapy: Immunotherapy and Targeted Drug Delivery.

This article reviews recent achievements in molecular immunology that have given a new insight into the mechanisms of tumor escape from the control of immunocompetent cells and opened the way to the development of radically new immunotherapeutic procedures and further improvement of existing chemotherapeutic techniques. The application of new updated techniques in molecular biology, biochemistry and cellular biotechnology has made it possible to detect molecular changes in tumor cells which help to escape them from immunological control and develop on their basis some novel approaches to immunotherapy of malignant tumors. One of the main challenges to immunotherapy, which is related to recognition of tumor antigens by immunocompetent cells, has been solved through generation of antitumor vaccines based on dendritic cells. The complexity of this procedure is in that these vaccines are strictly individual and must be prepared strictly individually for each concrete patient. However, high sensitivity of the novel procedure which makes it applicable even for patients with grades III and IV of cancer encourages hope that it will afford effective treatment and reduce metastatic growth, especially when used in combination with other concomitant immunochemotherapeutic techniques.

Journal Article↗

Targeted drug delivery.

Achieving adequate dosage of a drug to be effective, without crossing the line into toxicity, can be a problem. This paper reviews ongoing research into methods of delivering drugs only to diseased tissue or cells, minimally affecting surrounding tissue. In particular, the author discusses encapsulating drugs in liposomes (artificial phospholipid microcapsules) which will only bind to certain cells.

Drug Delivery Systems↗

Genetic modification of hematopoietic cells using retroviral and lentiviral vectors: safety considerations for vector design and delivery into target cells.

The recent development of leukemia in three patients following retroviral vector gene transfer in hematopoietic stem cells, resulting in the death of one patient, has raised safety concerns for the use of integrating gene transfer vectors for human gene therapy. This review discusses these serious adverse events from the perspective of whether restrictions on vector design and vector-modified target cells are warranted at this time. A case is made against presently establishing specific restrictions for vector design and transduced cells; rather, their safety should be ascertained by empiric evaluation in appropriate preclinical models on a case-by-case basis. Such preclinical data, coupled with proper informed patient consent and a risk-benefit ratio analysis, provide the best available prospective evaluation of gene transfer vectors prior to their translation into the clinic.

Genetic Therapy↗

Different approaches of gene therapy used in prostate cancer.

Prostate cancer is the second leading malignancy next to lung cancer among American males. Estimated number of new cases of prostate cancer by the year 2004 will be 230,110 and number of deaths due to this disease will be 29,900 (American Cancer Society, Surveillance Research, 2004). Major progress have been made in gene therapy due to significant advancement of biology including specialized fields like Molecular Biology, Cell Biology, Immunology and Molecular Virology which includes development of viral and non-viral vectors and delivery system, of which targeted delivery of suicide genes using tissue specific promoter-enhancer, combined therapy, gene replacement therapy are most notable. In this report, we will review about various aspects of gene therapy including most recent discoveries in this field.

Animals↗

[Adenovirus-mediated delivery of targeted ribonuclease against HBV replication in vitro].

OBJECTIVE: To observe the inhibitory effect of targeted ribonuclease delivered via adenovirus against HBV replication in vitro. METHODS: The shuttle plasmids pDC316 were constructed on the basis of the previous plasmids pcDNA3.1(-)/TRL, pcDNA3.1(-)/TR, pcDNA3.1(-)/TRmut, pcDNA3.1(-)/HBVc, and pcDNA3.1(-)/hEDN by subcloning the target gene sequences of TRL, TR, HBVc, and hEDN, respectively. HEK 293 cells were cotransfected with the pDC316 plasmids respectively in the presence of the rescue plasmid pBHGlox(delta)E1,3Cre to yield the recombinant adenoviral vectors which comprised the above genes. After transfection of HepG2.2.15 cells with the vectors, RAd/TRL expression was detected by indirect immunofluorescence staining and RT-PCR. Radioimmunoassay was used to analyse anti-HBV activity of RAd/TRL. RESULTS: Recombinant RAd vectors were prepared successfully. Effective expression of RAd/TRL in HepG2.2.15 cells resulted in a significant decrease of HBsAg and HBeAg concentration in comparison with the controls. CONCLUSION: Adenoviral vector-mediated targeted ribonuclease can effectively inhibit HBV replication.

Adenoviridae↗

Targeted sustained delivery of tobramycin at the site of a femoral osteotomy.

Complex fractures are difficult to manage because of the increased risk of secondary infection. Traditional treatments include debridment and local administration of antibiotics. Local antibiotic therapy is a safe technique resulting in high local concentration of antibiotics with minimal systemic levels. Local antibiotics effectively control infection in animal models. The length of implantation and the need for removal of the delivery vehicle places the patient at risk for additional surgical procedure as well as delays the fixation procedure. Development of a bioresorbable carrier that can deposit therapeutic concentrations of antibiotics locally without side-effects will provide positive outcomes for the patient. Tricalcium phosphate lysine (TCPL) ceramic capsules containing tobramycin were implanted at the site of a femoral osteotomy delivered therapeutic concentrations of the drug locally and reduced the incidence of infection compared to TCPL capsule which were uncharged by 50%. In addition, the ceramic material was osteoconductive and animals in TCPL + TOB and TCPL carrier alone showed evidence of osteoblast alkaline phosphatase activity for a period of 15 weeks. Neither the carrier nor the carrier containing antibiotics displayed untoward effects on body weight, vital organs and reproductive organs over a 15 week period. The results from this study demonstrated that TCPL can be used as an effective osteoconductive material capable of delivering therapeutic concentrations of antibiotics over 15 week period.

Animals↗

[Study on targeting drug delivery system--the animal behavior in vivo after hepatic arterial embolization of methotrexate microsphere].

The cross-linked MTX-gelatin microsphere was chosen as an experimental model. Pharmacokinetics and biodegradable period of the microsphere were studied after hepatic arterial embolization in vivo in dogs. After hepatic arterial infusion, the concentration of MTX-ms in peripheral vein blood increased rapidly and declined slowly. In contrast, the peak time of the drug concentration of infused MTX solution appeared at once and decreased rapidly. The half-life of MTX-ms in vivo was 2.14 times as much as that of MTX solution. During the assay period of sixteen hours the drug concentration of MTX-ms in hepatic vein blood was constantly higher than that of MTX solution. The follow-up angiography revealed that microsphere is a kind of peripheral embolic agent and its degradable time in vivo is about one month. Both sustained release and embolization effectiveness of microsphere will be significant in treatment of hepatic cancer.

Animals↗

[Study on targeting drug delivery system--the characteristics of methotrexate microsphere and experimental treatment of hepatic tumor in rats by arterial embolization].

Preparation of methotrexate microsphere (MTX-ms) by emulsion-freezing technique was introduced and the experimental results proved that MTX entrapped in the microspheres exhibited good stabilities towards temperature, cobalt-60 radiation and light. The dissolution and inflation rate of the microspheres in pH 7.4 buffer solution at different times measured by Coulter counter was presented. Antitumor activity of MTX-ms after hepatic arterial embolization was examined in a model of liver tumor in Wistar rats. The group of rats treated with MTX-ms showed a rather significant reduction in tumor growth and more extended tumor necrosis as compared with the other groups, e.g. normal saline solution, MTX solution, placebo gelatin-ms and the results demonstrate that the effect of arterial chemoembolization used by MTX-ms is superior to that of the groups either using arterial chemotherapy or arterial embolization alone in treating rat liver cancer.

Animals↗