Current Issue : October - December Volume : 2017 Issue Number : 4 Articles : 6 Articles
Background: As cancer metastasis is the deadliest aspect of cancer, causing 90% of human deaths, evaluating the\nmolecular mechanisms underlying this process is the major interest to those in the drug development field. Both\ntherapeutic target identification and proof-of-concept experimentation in anti-cancer drug development require\nappropriate animal models, such as xenograft tumor transplantation in transgenic and knockout mice. In the\nprogression of cancer metastasis, circulating tumor cells (CTCs) are the most critical factor in determining the\nprognosis of cancer patients. Several studies have demonstrated that measuring CTC-specific markers in a clinical setting\n(e.g., flow cytometry) can provide a current status of cancer development in patients. However, this useful technique has\nrarely been applied in the real-time monitoring of CTCs in preclinical animal models.\nMethods: In this study, we designed a rapid and reliable detection method by combining a bioluminescent in vivo\nimaging system (IVIS) and quantitative polymerase chain reaction (QPCR)-based analysis to measure CTCs in\nanimal blood. Using the IVIS Spectrum CT System with 3Dââ?¬â??imaging on orthotropic-developed breast-tumor-bearing mice.\nResults: In this manuscript, we established a quick and reliable method for measuring CTCs in a preclinical animal mode.\nThe key to this technique is the use of specific human and mouse GUS primers on DNA/RNA of mouse peripheral blood\nunder an absolute qPCR system. First, the high sensitivity of cancer cell detection on IVIS was presented by measuring the\nluciferase carried MDA-MB-231 cells from 5 to 5x1011 cell numbers with great correlation (R2 = 0.999). Next, the MDA-MB-\n231 cell numbers injected by tail vein and their IVIS radiance signals were strongly corrected with qPCR-calculated copy\nnumbers (R2 > 0.99). Furthermore, by applying an orthotropic implantation animal model, we successfully distinguished\nxenograft tumor-bearing mice and control mice with a significant difference (p < 0.001), whereas IVIS Spectrum-CT 3Dââ?¬â??\nvisualization showed that blood of mice with lung metastasis contained more than twice the CTC numbers than ordinary\ntumor-bearing mice. We demonstrated a positive correlation between lung metastasis status and CTC numbers\nin peripheral mouse blood.\nConclusion: Collectively, the techniques developed for this study resulted in the integration of CTC assessments into\npreclinical models both in vivo and ex vivo, which will facilitate translational targeted therapy in clinical practice....
Background: Tetanus-reduced dose diphtheria-acellular pertussis (Tdap) vaccination during adolescence was\nintroduced in response to the resurgence of pertussis in various countries. A new Tdap vaccine was manufactured\nin Korea as a countermeasure against a predicted Tdap vaccine shortage. This study was performed to evaluate the\nimmunogenicity, safety, and protection efficacy against Bordetella pertussis of the new Tdap vaccine in a murine\nmodel.\nMethods: Four-week-old BABL/c mice were used for assessment of immunogenicity and protection efficacy. A\nsingle dose of primary diphtheria-tetanus-acellular pertussis (DTaP) vaccine was administered, followed by a single\ndose of Tdap booster vaccine after a 12-week interval. Anti-pertussis toxin (PT), anti-filamentous hemagglutinin\n(FHA), and anti-pertactin (PRN) IgG titers were measured before primary vaccination, and before and after booster\nvaccination. An intranasal challenge test was performed after booster vaccination to determine protection efficacy.\nTo assess safety, mouse weight gain test and leukocytosis promotion test were performed using 4-week-old ddY\nfemale mice.\nResults: Anti-PT and anti-FHA IgG titers after booster vaccination were significantly higher than those before\nbooster vaccination with either the new vaccine or a commercially available Tdap vaccine (P = 0.01 for all\noccasions). After booster vaccination, no significant difference was observed between the two vaccines in antibody\ntiters against pertussis antigens (P = 0.53 for anti-PT IgG, P = 0.91 for anti-FHA IgG, P = 0.39 for anti-PRN IgG). In the\nintranasal challenge test, inoculated B. pertussis was eradicated 7 days after infection. On days 4 and 7 after\ninfection, colony counts of B. pertussis were not significantly different between the new and positive control\nvaccine groups (P = 1.00). Mean body weight changes and leukocyte counts of the new vaccine, positive control,\nand negative control groups were not significantly different 7 days after vaccination (P = 0.87 and P = 0.37,\nrespectively). All leukocyte counts in the new vaccine group were within a mean �± 3 standard deviations range. Conclusions: A murine model involving a single dose primary DTaP vaccination followed by a single dose Tdap\nbooster vaccination can be used for non-clinical studies of Tdap vaccines. The new Tdap vaccine manufactured in\nKorea exhibited comparable immunogenicity, protection efficacy, and safety with a commercially available Tdap\nvaccine....
Bee venom has special pharmacological activity for its enzymes and peptides containment. Melittin is the bee venom’s main constituent. The present work aimed in-vitro estimation of the anti-cancer potentials of bee venom (BV) in addition to evaluation of the synergetic potential of dates extract to bee venom against lung cancer (A549). BV showed a higher toxicity to A549 cells accompanied with synergetic activity of its toxicity in combination with dates extracts where the IC50 was arranged in the order of BV> Mix> Dates compared with doxorubicin as a positive control. Data revealed that BV/dates extract has a synergetic potential to BV that was accompanied with anticancer activity via up/down regulation of pro and anti apoptotic genes compared with control; caspase-3, Bcl-2 and pro-apoptotic gene (p53). Cell cycle analysis revealed that bee venom, dates and their mix induced pre G1 apoptosis and cell cycle arrest at G2/M phase....
Background: Even though hematopoietic stem cell transplantation can be curative in patients with severe\ncombined immunodeficiency, there is a need for additional strategies boosting T cell immunity in individuals\nsuffering from genetic disorders of lymphoid development. Here we show that image-guided intrathymic injection\nof hematopoietic stem and progenitor cells in NOD-scid IL2r�³null mice is feasible and facilitates the generation of\nfunctional T cells conferring protective immunity.\nMethods: Hematopoietic stem and progenitor cells were isolated from the bone marrow of healthy C57BL/6 mice\n(wild-type, Luciferase+, CD45.1+) and injected intravenously or intrathymically into both male and female, young or\naged NOD-scid IL2r�³null recipients. The in vivo fate of injected cells was analyzed by bioluminescence imaging and\nflow cytometry of thymus- and spleen-derived T cell populations. In addition to T cell reconstitution, we evaluated\nmice for evidence of immune dysregulation based on diabetes development and graft-versus-host disease. T cell\nimmunity following intrathymic injection of hematopoietic stem and progenitor cells in NOD-scid IL2r�³null mice\nwas assessed in a B cell lymphoma model.\nResults: Despite the small size of the thymic remnant in NOD-scid IL2r�³null mice, we were able to accomplish\nprecise intrathymic delivery of hematopoietic stem and progenitor cells by ultrasound-guided injection.\nThymic reconstitution following intrathymic injection of healthy allogeneic hematopoietic cells was most\neffective in young male recipients, indicating that even in the setting of severe immunodeficiency, sex and\nage are important variables for thymic function. Allogeneic T cells generated in intrathymically injected NOD-scid\nIL2r�³null mice displayed anti-lymphoma activity in vivo, but we found no evidence for severe auto/alloreactivity in\nT cell-producing NOD-scid IL2r�³null mice, suggesting that immune dysregulation is not a major concern.\nConclusions: Our findings suggest that intrathymic injection of donor hematopoietic stem and progenitor cells is a\nsafe and effective strategy to establish protective T cell immunity in a mouse model of severe combined\nimmunodeficiency....
Orange Albedo (OW/OE) and grape seed extracts (GW/GE) were considered for their medical potentials against carcinoma cells due to their bioflavonoid derivatives. Cytotoxic activity of the OA and GSEs were investigated against breast (MCF-7) and colorectal (HCT-116) cancer cell lines using MTT assay. Moreover, cell cycle distribution by flow cytometry and RT-PCR were conducted. Data recorded revealed that the IC50 value of OW/OE were 1.50 mg/ml and 14.57 mg/ml respectively against MCF-7 cells, 0.50 mg/ml and 0.88 mg/ml respectively against HCT-116 cells. While the IC50 values of GW/GE against MCF-7 were 1.36 mg/ml and 0.17 mg/ml respectively, HCT-116 cancer cell line were 0.21 mg/ml and 0.80 mg/ml respectively. Flow-cytometry analysis showed MCF-7 arrest in the G2/M phase and S phase post treatment with OW/OE. Both OW/OE and GW/GE exert their effects in MCF-7 and HCT-116 cell lines via Bax, p53 and Casp3 up-regulation and Bcl-2 down regulation in a dose dependent manner. These observations suggest the hypothesis that OW/OE and GW/GE could be used as supplementary medication to the current one against breast and colorectal cancers due to their promising effects on cellular proliferation inhibition and apoptosis induction....
Background: Due to the multilineage differentiation ability and paracrine role of adipose-derived stem cells (ASCs)\nfor bladder defect repair, various scaffolds have been applied in combination with ASCs to promote bladder\nregeneration and restore bladder function. However, the low survival rate of ASCs and the difficulty of promoting\nbladder functional recovery are still unsolved. To explore these problems, we investigated the feasibility of a novel\nscaffold seeded with ASCs in a rat model of bladder augmentation.\nMethods: A novel autologous myofibroblast (AM)-silk fibroin (SF) scaffold was harvested after subcutaneously\nprefabricating the bladder acellular matrix grafts (BAMG) and SF by removing the BAMG. The AM-SF scaffolds were\nthen seeded with ASCs (AM-SF-ASCs). Fifty percent supratrigonal cystectomies were performed followed by\naugmenting the cystectomized defects with AM-SF scaffolds or AM-SF-ASCs. The histological and functional\nassessments of bladders were performed 2, 4, and 12 weeks after surgery while the ASCs were tracked in vivo.\nResults: For bladder tissue regeneration, immunofluorescence analysis revealed that AM-SF-ASCs (the experimental\ngroup) promoted better morphological regeneration of the urothelium, vessels, bladder smooth muscle, and nerve\nthan AM-SF scaffolds (the control group). Regarding functional restoration, the AM-SF-ASC group exhibited higher\nbladder compliance and relatively normal micturition pattern compared to the AM-SF group. In addition, a certain\nnumber of surviving ASCs could be found in vivo 12 weeks after implantation, and some of them had differentiated\ninto smooth muscle cells.\nConclusions: The AM-SF scaffolds with ASCs could rapidly promote bladder morphological regeneration and\nimproved bladder urinary function. In addition, the bag-shaped structure of the AM-SF scaffold can improve the\nsurvival of ASCs for at least 12 weeks. This strategy of AM-SF-ASCs has a potential to repair large-scale bladder\ndefects in the clinic in the future....
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