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      1. Author :
        Nguyen, Leslie; Zhong, Wei-Zhu; Painter, Cory L; Zhang, Cathy; Rahavendran, Sadayappan V; Shen, Zhongzhou
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        Journal of pharmaceutical and biomedical analysis
      6. Products :
      7. Volume :
        53
      8. Issue :
        3
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; Bioware; Chromatography, Liquid; Cyclin-Dependent Kinase 4; Drug Stability; Female; Humans; MDA-MB-231-D3H1 cells; Mice; Mice, SCID; Neoplasm Transplantation; Neoplasms, Experimental; Piperazines; Protein kinase inhibitors; Pyridines; Sensitivity and Specificity; Tandem Mass Spectrometry; Transplantation, Heterologous
      12. Abstract :
        Phase II attrition of clinical candidates in the drug development cycle is currently a major issue facing the pharmaceutical industry. To decrease phase II attrition, there is an increased emphasis on validation of mechanism of action, development of efficacy models and measurement of drug levels at the site of action. PD 0332991, a highly specific inhibitor of cyclin-dependent kinase 4 (CDK-4) is currently in clinical development for the treatment of solid tumor. A clinical presurgical study will be required to better understand how PD 0332991 affects signaling pathways and how the intratumoral concentration of PD 0332991 correlates with plasma PK parameters and molecular alterations in breast cancer tissues after PD 0332991 treatment. Before conducting such a clinical study, it is important to evaluate PD 0332991 levels in tumor tissue samples from a xenograft mouse model for the determination of drug exposure at the site of action. Therefore, the objectives of this study were (1) to develop and validate a sensitive LC-MS/MS method to quantify PD 0332991 in mouse tumor tissues from MDA-MB-231-Luc human breast tumor xenografts in SCID-beige mice; (2) to quantify PD 0332991 levels in mouse tumor tissues after oral administration of PD 0332991 at 10 and 100mg/kg using the validated LC-MS/MS method. Both liquid-liquid extraction (LLE) and supported liquid extraction (SLE) in a 96-well format were developed and evaluated to achieve optimal extraction recovery with minimal matrix effects. The newly developed SLE method is more efficient (speed and ease) and demonstrates comparable recovery (93.1-100% at three different concentrations) compared to the traditional LLE method. The validated LC-MS/MS for PD 032291 in mouse tumor tissue homogenate method exhibited a linear dynamic range of 0.1-100 ng/mL with inter-day accuracy and precision within 15%. The validated method was successfully applied to measure PD 0332991 levels in tumor tissues in MDA-MB-231-Luc human breast tumor xenografts in SCID beige mice. The mean tumor concentrations at 6h post-oral PD 0332991 administration at 10 and 100mg/kg were 1793 (+/-1008) and 25,163 (+/-3959) ng/g, respectively.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/20236782
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8987
      1. Author :
        Li, Min; Rigby, Kevin; Lai, Yuping; Nair, Vinod; Peschel, Andreas; Schittek, Birgit; Otto, Michael
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Antimicrobial agents and chemotherapy
      6. Products :
      7. Volume :
        53
      8. Issue :
        10
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Anti-Bacterial Agents; Bioware; Blotting, Southern; Chromatography, Thin Layer; Computational Biology; Cytochromes c; Genetic Complementation Test; Humans; Microscopy, Electron, Scanning; Microscopy, Immunoelectron; Mutagenesis; Peptides; Phospholipids; Polymerase Chain Reaction; Staphylococcus aureus; Xen36
      12. Abstract :
        Antimicrobial peptides (AMPs) form an important part of the innate host defense. In contrast to most AMPs, human dermcidin has an anionic net charge. To investigate whether bacteria have developed specific mechanisms of resistance to dermcidin, we screened for mutants of the leading human pathogen, Staphylococcus aureus, with altered resistance to dermcidin. To that end, we constructed a plasmid for use in mariner-based transposon mutagenesis and developed a high-throughput cell viability screening method based on luminescence. In a large screen, we did not find mutants with strongly increased susceptibility to dermcidin, indicating that S. aureus has no specific mechanism of resistance to this AMP. Furthermore, we detected a mutation in a gene of unknown function that resulted in significantly increased resistance to dermcidin. The mutant strain had an altered membrane phospholipid pattern and showed decreased binding of dermcidin to the bacterial surface, indicating that dermcidin interacts with membrane phospholipids. The mode of this interaction was direct, as shown by assays of dermcidin binding to phospholipid preparations, and specific, as the resistance to other AMPs was not affected. Our findings indicate that dermcidin has an exceptional value for the human innate host defense and lend support to the idea that it evolved to evade bacterial resistance mechanisms targeted at the cationic character of most AMPs. Moreover, they suggest that the antimicrobial activity of dermcidin is dependent on the interaction with the bacterial membrane and might thus assist with the determination of the yet unknown mode of action of this important human AMP.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/19596877
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        9983
      1. Author :
        Pozo, J. L. del; Rouse, M. S.; Mandrekar, J. N.; Sampedro, M. F.; Steckelberg, J. M.; Patel, R.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Antimicrobial Agents and Chemotherapy
      6. Products :
      7. Volume :
        53
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen, Xen30, Xen5, Xen41
      12. Abstract :
        Bacterial biofilms are resistant to conventional antimicrobial agents. Prior in vitro studies have shown that electrical current (EC) enhances the activities of aminoglycosides, quinolones, and oxytetracycline against Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus epidermidis, Escherichia coli, and Streptococcus gordonii. This phenomenon, known as the bioelectric effect, has been only partially defined. The purpose of this work was to study the in vitro bioelectric effect on the activities of 11 antimicrobial agents representing a variety of different classes against P. aeruginosa, methicillin-resistant Staphylococcus aureus (MRSA), and S. epidermidis. An eight-channel current generator/controller and eight chambers delivering a continuous flow of fresh medium with or without antimicrobial agents and/or EC to biofilm-coated coupons were used. No significant decreases in the numbers of log10 CFU/cm2 were seen after exposure to antimicrobial agents alone, with the exception of a 4.57-log-unit reduction for S. epidermidis and trimethoprim-sulfamethoxazole. We detected a statistically significant bioelectric effect when vancomycin plus 2,000 microamperes EC were used against MRSA biofilms (P = 0.04) and when daptomycin and erythromycin were used in combination with 200 or 2,000 microamperes EC against S. epidermidis biofilms (P = 0.02 and 0.0004, respectively). The results of these experiments indicate that the enhancement of the activity of antimicrobial agents against biofilm organisms by EC is not a generalizable phenomenon across microorganisms and antimicrobial agents.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/18725436
      14. Call Number :
        137347
      15. Serial :
        5991
      1. Author :
        Pozo, J. L. del; Rouse, M. S.; Mandrekar, J. N.; Steckelberg, J. M.; Patel, R.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Antimicrob Agents Chemother
      6. Products :
      7. Volume :
        53
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Aza Compounds/pharmacology, Biofilms/drug effects/*growth & development, Electricity/*adverse effects, Pseudomonas/drug effects/*growth & development, Quinolines/pharmacology, Staphylococcus/drug effects/*growth & development, Tobramycin/pharmacology IVIS, Xenogen, Xen30
      12. Abstract :
        The activity of electrical current against planktonic bacteria has previously been demonstrated. The short-term exposure of the bacteria in biofilms to electrical current in the absence of antimicrobials has been shown to have no substantial effect; however, longer-term exposure has not been studied. A previously described in vitro model was used to determine the effect of prolonged exposure (i.e., up to 7 days) to low-intensity (i.e., 20-, 200-, and 2,000-microampere) electrical direct currents on Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis biofilms. Dose- and time-dependent killing was observed. A maximum of a 6-log(10)-CFU/cm(2) reduction was observed when S. epidermidis biofilms were exposed to 2,000 microamperes for at least 2 days. A 4- to 5-log(10)-CFU/cm(2) reduction was observed when S. aureus biofilms were exposed to 2,000 microamperes for at least 2 days. Finally, a 3.5- to 5-log(10)-CFU/cm(2) reduction was observed when P. aeruginosa biofilms were exposed to electrical current for 7 days. A higher electrical current intensity correlated with greater decreases in viable bacteria at all time points studied. In conclusion, low-intensity electrical current substantially reduced the numbers of viable bacteria in staphylococcal or Pseudomonas biofilms, a phenomenon we have labeled the “electricidal effect.”
      13. URL :
        http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18955534
      14. Call Number :
        137350
      15. Serial :
        7845
      1. Author :
        Dai, T.; Tegos, G. P.; Burkatovskaya, M.; Castano, A. P.; Hamblin, M. R.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2009
      5. Publication :
        Antimicrobial Agents and Chemotherapy
      6. Products :
      7. Volume :
        53
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, Xenogen, Xen5, Xen44
      12. Abstract :
        An engineered chitosan acetate bandage preparation (HemCon) is used as a hemostatic dressing, and its chemical structure suggests that it should also be antimicrobial. We previously showed that when a chitosan acetate bandage was applied to full-thickness excisional wounds in mice that had been infected with pathogenic bioluminescent bacteria (Pseudomonas aeruginosa, Proteus mirabilis, and Staphylococcus aureus), it was able to rapidly kill the bacteria and save the mice from developing fatal infections. Wound healing was also stimulated. In the present study, we asked whether a chitosan acetate bandage could act as a topical antimicrobial dressing when it was applied to third-degree burns in mice contaminated with two of these bacterial species (P. aeruginosa and P. mirabilis). Preliminary experiments established the length of burn time and the number of bacteria needed to produce fatal infections in untreated mice and established that the chitosan acetate bandage could adhere to the infected burn for up to 21 days. In the case of P. aeruginosa infections, the survival rate of mice treated with the chitosan acetate bandage was 73.3% (whereas the survival rate of mice treated with a nanocrystalline silver dressing was 27.3% [P = 0.0055] and that of untreated mice was 13.3% [P < 0.0002]). For P. mirabilis infections, the comparable survival rates were 66.7%, 62.5%, and 23.1% respectively. Quantitative bioluminescent signals showed that the chitosan acetate bandage effectively controlled the growth of bacteria in the burn and prevented the development of systemic sepsis, as shown by blood culture. These data suggest that chitosan acetate bandage is efficacious in preventing fatal burn infections.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/19015341
      14. Call Number :
        137209
      15. Serial :
        5713
      1. Author :
        Ragas, X.; Sanchez-Garcia, D.; Ruiz-Gonzalez, R.; Dai, T.; Agut, M.; Hamblin, M. R.; Nonell, S.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        J Med Chem
      6. Products :
      7. Volume :
        53
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Xen31, Xen 31, MRSA, S. aureus, IVIS, Bioluminescence, Animals; Bacterial Infections/*drug therapy; Burns/drug therapy/microbiology; Candida/drug effects; Cations; Gram-Negative Bacteria/drug effects; Gram-Positive Bacteria/drug effects; Male; Methicillin-Resistant Staphylococcus aureus; Mice; Mice, Inbred BALB C; *Photochemotherapy; Photosensitizing Agents/*chemical synthesis/chemistry/pharmacology; Porphyrins/*chemical synthesis/chemistry/pharmacology; Solubility; Staphylococcal Infections/drug therapy/microbiology; Structure-Activity Relationship
      12. Abstract :
        Structures of typical photosensitizers used in antimicrobial photodynamic therapy are based on porphyrins, phthalocyanines, and phenothiazinium salts, with cationic charges at physiological pH values. However, derivatives of the porphycene macrocycle (a structural isomer of porphyrin) have barely been investigated as antimicrobial agents. Therefore, we report the synthesis of the first tricationic water-soluble porphycene and its basic photochemical properties. We successfully tested it for in vitro photoinactivation of different Gram-positive and Gram-negative bacteria, as well as a fungal species (Candida) in a drug-dose and light-dose dependent manner. We also used the cationic porphycene in vivo to treat an infection model comprising mouse third degree burns infected with a bioluminescent methicillin-resistant Staphylococcus aureus strain. There was a 2.6-log(10) reduction (p < 0.001) of the bacterial bioluminescence for the PDT-treated group after irradiation with 180 J.cm(-2) of red light.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/20936792
      14. Call Number :
        PKI @ kd.modi @ 1
      15. Serial :
        10555
      1. Author :
        Ragas, X.; Sanchez-Garcia, D.; Ruiz-Gonzalez, R.; Dai, T.; Agut, M.; Hamblin, M. R.; Nonell, S.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        J Med Chem
      6. Products :
      7. Volume :
        53
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Xen31, Xen 31, MRSA, S. aureus, IVIS, Bioluminescence, Animals; Bacterial Infections/*drug therapy; Burns/drug therapy/microbiology; Candida/drug effects; Cations; Gram-Negative Bacteria/drug effects; Gram-Positive Bacteria/drug effects; Male; Methicillin-Resistant Staphylococcus aureus; Mice; Mice, Inbred BALB C; *Photochemotherapy; Photosensitizing Agents/*chemical synthesis/chemistry/pharmacology; Porphyrins/*chemical synthesis/chemistry/pharmacology; Solubility; Staphylococcal Infections/drug therapy/microbiology; Structure-Activity Relationship
      12. Abstract :
        Structures of typical photosensitizers used in antimicrobial photodynamic therapy are based on porphyrins, phthalocyanines, and phenothiazinium salts, with cationic charges at physiological pH values. However, derivatives of the porphycene macrocycle (a structural isomer of porphyrin) have barely been investigated as antimicrobial agents. Therefore, we report the synthesis of the first tricationic water-soluble porphycene and its basic photochemical properties. We successfully tested it for in vitro photoinactivation of different Gram-positive and Gram-negative bacteria, as well as a fungal species (Candida) in a drug-dose and light-dose dependent manner. We also used the cationic porphycene in vivo to treat an infection model comprising mouse third degree burns infected with a bioluminescent methicillin-resistant Staphylococcus aureus strain. There was a 2.6-log(10) reduction (p < 0.001) of the bacterial bioluminescence for the PDT-treated group after irradiation with 180 J.cm(-2) of red light.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/20936792
      14. Call Number :
        PKI @ kd.modi @ 5
      15. Serial :
        10556
      1. Author :
        Luo, Z R; Huang, T; Li, W; Shen, B Z
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        Panminerva medica
      6. Products :
      7. Volume :
        52
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Animals; B16-F10-luc-G5 cells; Bioware; Diagnostic Imaging; Luminescence; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Molecular Dynamics Simulation
      12. Abstract :
        AIM The aim of this study was to evaluate the veracity and sensitivity of in-vivo imaging system (IVIS) for inspection of tumor dynamic morphology. METHODS Mouse melanoma cells (B16-F10-luc-G5) in 100 mL media were seeded into a 96-well plate by 1:2 serial dilution from 10000 cells (well #1) to 78 cells (well #8). The plate was imaged using IVIS system to evaluate its sensitivity for luminescence. Ten Bablc mice with tumor cells were injected subcutaneously (1 x 10(5) in 100 mL) and tumor luminescence was detected by IVIS at Day 0, Day 3, Day 5, Day 7 and Day 9. RESULTS As few as 78 tumor cells were detectable by IVIS. A strong correlation between number of tumor cells and bioluminescence (R2=0.99) was also demonstrated. Tumor luminescence were observed in all mice by IVIS at all days, and there was significant difference (P<0.01) between each two days from Day 0 to Day 9. Moreover, tumor dynamic morphology could be monitored by IVIS when it is invisible. CONCLUSION Compared with conventional methods, with high veracity and sensitivity, IVIS system should be recommended as an effective method for inspection of tumor dynamic morphology.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/20228722
      14. Call Number :
        PKI @ catherine.lautenschlager @
      15. Serial :
        8995
      1. Author :
        Adachi, T.; Kawakami, E.; Ishimaru, N.; Ochiya, T.; Hayashi, Y.; Ohuchi, H.; Tanihara, M.; Tanaka, E.; Noji, S.
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2010
      5. Publication :
        Dev Growth Differ
      6. Products :
      7. Volume :
        52
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS, B16-F10-luc-G5, B16F10-luc-G5, B16-F10-luc, B16F10-luc, Animals; Base Sequence; Cell Line, Tumor; Collagen/*chemistry; DNA Primers; *Gene Silencing; Mice; RNA, Small Interfering/*administration & dosage/*chemistry; Reverse Transcriptase Polymerase Chain Reaction
      12. Abstract :
        Silencing gene expression by small interfering RNAs (siRNAs) has become a powerful tool for the genetic analysis of many animals. However, the rapid degradation of siRNA and the limited duration of its action in vivo have called for an efficient delivery technology. Here, we describe that siRNA complexed with a synthetic collagen poly(Pro-Hyp-Gly) (SYCOL) is resistant to nucleases and is efficiently transferred into cells in vitro and in vivo, thereby allowing long-term gene silencing in vivo. We found that the SYCOL-mediated local application of siRNA targeting myostatin, coding a negative regulator of skeletal muscle growth, in mouse skeletal muscles, caused a marked increase in the muscle mass within a few weeks after application. Furthermore, in vivo administration of an anti-luciferase siRNA/SYCOL complex partially reduced luciferase expression in xenografted tumors in vivo. These results indicate a SYCOL-based non-viral delivery method could be a reliable simple approach to knockdown gene expression by RNAi in vivo as well as in vitro.
      13. URL :
        http://www.ncbi.nlm.nih.gov/pubmed/20874713
      14. Call Number :
        PKI @ kd.modi @ 11
      15. Serial :
        10352
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