Level Of Growth Inhibition Research Articles

A Novel Method for Fresh-Cut Decontamination: Efficiency of Vaporized Ethyl Pyruvate in Reducing S taphylococcus aureus and E scherichia coli O157:H7 from Fresh Parsley

Vaporized ethyl pyruvate (EP) was evaluated for its efficiency for decontamination of parsley from Escherichia coli O157:H7 and Staphylococcus aureus. Inoculated parsleys (∼104 or ∼106 cfu/g) were treated with 0 (control sample), 100, 400 and 1,000 μL of vaporized EP in 2.6-L closed-lid food containers. Samples were evaluated for bacterial growth and visual acceptance during treatment. In the case of decontamination of E. coli O157:H7, 1,000 μL EP treatments completely inhibited the bacterial population and enabled 100% growth inhibition levels (GILs) in both inoculum levels. In low inoculation of S. aureus, the difference between the counts of the control and 1,000 μL EP-treated samples was higher than 2 logs after the storage. In high inoculum, 400 or 1,000 μL EP-treated parsleys had significantly (P < 0.05) lower S. aureus populations as compared with the control sample. EP treatment retarded parsley decay; however, it caused some yellowness on the parsley leaves. Practical Applications Safety of minimally processed fruits and vegetables is of great concern worldwide as no decontamination treatment is included in their processing. Different methods such as washing with chemical sanitizers, pulsed ultraviolet light, high pressure and ionized radiation have been tested for decontamination of fresh products. In general, these methods achieve less than a 2-log reduction and negatively affect organoleptical properties of the products. Surface structure of the product also influences the effectiveness of surface decontamination processes as pathogens may penetrate into protected sites within plant tissue. Vaporized ethyl pyruvate (EP) has potential to remove these disadvantages because of its volatility which enables it to reach the pathogens that are hidden within plant structures. In this study, EP was effective against both Escherichia coli O157:H7 and Staphylococcus aureus on fresh parsley. As a conclusion, EP was confirmed as an effective agent for decontamination of parsleys and as a successful alternative for other decontamination methods.

The Dilemma of Antimicrobial Treatment of Shiga Toxin-producing Escherichia coli

The Dilemma of Antimicrobial Treatment of Shiga Toxin-producing Escherichia coli

A novel ATM/TP53/p21-mediated checkpoint only activated by chronic γ-irradiation.

Different levels or types of DNA damage activate distinct signaling pathways that elicit various cellular responses, including cell-cycle arrest, DNA repair, senescence, and apoptosis. Whereas a range of DNA-damage responses have been characterized, mechanisms underlying subsequent cell-fate decision remain elusive. Here we exposed cultured cells and mice to different doses and dose rates of γ-irradiation, which revealed cell-type-specific sensitivities to chronic, but not acute, γ-irradiation. Among tested cell types, human fibroblasts were associated with the highest levels of growth inhibition in response to chronic γ-irradiation. In this context, fibroblasts exhibited a reversible G1 cell-cycle arrest or an irreversible senescence-like growth arrest, depending on the irradiation dose rate or the rate of DNA damage. Remarkably, when the same dose of γ-irradiation was delivered chronically or acutely, chronic delivery induced considerably more cellular senescence. A similar effect was observed with primary cells isolated from irradiated mice. We demonstrate a critical role for the ataxia telangiectasia mutated (ATM)/tumor protein p53 (TP53)/p21 pathway in regulating DNA-damage-associated cell fate. Indeed, blocking the ATM/TP53/p21 pathway deregulated DNA damage responses, leading to micronucleus formation in chronically irradiated cells. Together these results provide insights into the mechanisms governing cell-fate determination in response to different rates of DNA damage.

In vitro testing of drug combinations employing nilotinib and alkylating agents with regard to pretransplant conditioning treatment of advanced-phase chronic myeloid leukemia.

The prognosis of patients with advanced-phase chronic myeloid leukemia (CML) remains dismal despite the availability of targeted therapies and allogeneic stem cell transplantation (allo-SCT). Increasing the antileukemic efficacy of the pretransplant conditioning regimen may be a strategy to increase remission rates and duration. We therefore investigated the antiproliferative effects of nilotinib in combination with drugs that are usually used for conditioning: the alkylating agents mafosfamide, treosulfan, and busulfan. Drug combinations were tested in vitro in different imatinib-sensitive and imatinib-resistant BCR-ABL-positive cell lines. A tetrazolium-based MTT assay was used for the assessment and quantification of growth inhibition after exposure to alkylating agents alone or to combinations with nilotinib. Drug interaction was analyzed using the median-effect method of Chou and Talalay, and combination index (CI) values were calculated according to the classic isobologram equation. Treatment of imatinib-sensitive, BCR-ABL-positive K562 and LAMA84 cells with nilotinib in combination with mafosfamide, treosulfan, or busulfan resulted in synergistic (CI < 1), additive (CI ~ 1), and predominantly antagonistic (CI > 1) effects, respectively. In imatinib-resistant K562-R and LAMA84-R cells, all applied drug combinations were synergistic (CI < 1) at higher growth inhibition levels. Our in vitro data warrant further investigation and may provide the basis for nilotinib-supplemented conditioning regimens for allo-SCT in advanced-phase CML.

Autophagy-mediated growth inhibition of malignant glioma cells by the BH3-mimetic gossypol

A major concern with regard to glioma treatment arises from the fact that high-grade gliomas are insensitive to the majority of anticancer therapies. The aim of the present study was to investigate antiproliferation potential of the BH3-mimetic gossypol in four glioma cells with different BRAF mutation status. Gossypol induced similar levels of growth inhibition in all glioma cell lines regardless of the BRAF mutation status. Glioma cells were found to be resistant to gossypol-mediated apoptosis; gossypol caused only a weak increase in the number of G2/M cells. However, gossypol treatment of glioma cells resulted in the progressive emergence of autophagic cells. In addition, the autophagy inhibitor 3-methyladenine (3-MA) partially rescued the growth inhibitory effect of gossypol on DBTRG-05MG cells, suggesting that pro-autophagic processes may be one of the underlying mechanisms for the sensitization of tumor cells to gossypol. Our data suggest that gossypol contributes to a more effective therapeutic strategy for brain tumor patients in which activation of apoptosis does not occur.

Autophagy‐mediated growth inhibition of BH3‐mimetic gossypol in malignant glioma cells (796.2)

In the present study, we evaluated the individual IC50 values of a BH3‐mimetic gossypol for its cell growth inhibitory effects in four glioma cell lines with different BRAF mutational statuses. NMC‐G1, and DBTRG‐05MG cell lines are heterozygous for BRAF‐V600E; T98G, and U‐87MG cell lines are homozygous for wild‐type (WT) BRAF. Gossypol induced similar levels of growth inhibition in all of glioma cell lines. However, gossypol exhibited no significant effect on the phosphorylation of MEK‐ERK in all three cell lines regardless of both BRAF mutation and drug resistance, implying that gossypol activity is independent of MEK‐ERK inhibition. Glioma cells were found to be resistant to the apoptosis mediated by gossypol, which caused only a weak increase in G2/M cells. Rather, gossypol resulted in the progressive emergence of autophagic cells in glioma cells, as determined by GFP‐LC3 puncta/cell counts. Further, autophagy inhibition with 3‐methyladenine partially rescued the growth inhibitory effect of gossypol in DBTRG‐05MG cells, suggesting that the pro‐death autophagic processes may be one of the underlying mechanisms for the sensitization of tumor cells toward gossypol. Collectively, our data suggest that gossypol may contribute to a more effective therapeutic strategy for brain tumor patients that are unable to activate apoptosis.

Autophagy prevents doxorubicin-induced apoptosis in osteosarcoma

Autophagy is a process of selective degradation of cellular components. Autophagy is an adaptive process in the majority of tumor cells; it provides sufficient nutrients by degrading cellular components to enhance the survival of tumors. Osteosarcoma is the most common type of primary malignant bone tumor in children and adolescents. Identification of an improved therapeutic strategy for the treatment of osteosarcoma is urgently required. Osteosarcoma has been primarily treated by chemotherapy and the phenomena of resistance to the therapy has become increasingly common. Doxorubicin (Dox) is a classic chemotherapeutic drug for the treatment of osteosarcoma, and certain studies have suggested that Dox induces autophagy. On the basis of the protective effect of autophagy for tumors, the present study investigated whether U2OS and Saos-2 osteosarcoma cells activate autophagy to reduce Dox-induced apoptosis. Dox was observed to inhibit the growth of U2OS and Saos-2 osteosarcoma cells in a concentration-dependent manner. The results of the western blot analysis demonstrated that Dox induced increased expression levels of the apoptosis-related proteins cleaved caspase-3 and cytochrome c and loss of mitochondrial membrane potential (MMP) in the U2OS and Saos-2 osteosarcoma cells. Furthermore, the results of the western blot analysis also revealed that Dox increased the expression levels of the autophagy-related protein microtubule-associated protein 1 light chain 3 and reduced those of p62 in the U2OS and Saos-2 osteosarcoma cells. In order to determine the effect of autophagy on the apoptosis induced by Dox in the U2OS and Saos-2 osteosarcoma cells, autophagy-related protein (Atg)7 small interfering (si) RNA or the autophagy inhibitor 3-methyladenine (3-MA) alone or combined with Dox was used in U2OS and Saos-2 osteosarcoma cells. The results identified that Atg7 siRNA and the autophagy inhibitor 3-MA significantly elevated the levels of growth inhibition by Dox and markedly increased the expression levels of the apoptosis‑related proteins cleaved caspase-3 and cytochrome c, and reduced the levels of MMP in the U2OS and Saos-2 osteosarcoma cells, which were treated with Dox. These results indicated that autophagy was the protective mechanism used by U2OS and Saos-2 osteosarcoma against Dox-induced apoptosis. The inhibition of autophagy notably increases the levels of apoptosis induced by Dox. This suggested that Dox used in combination with autophagy inhibitors may effectively treat osteosarcoma.

Influence of magnesium on copper phytotoxicity to and accumulation and translocation in grapevines

The phytotoxic effects of excess copper (Cu) on grapevines (Vitis vinifera L. var. Kyoho) were examined, both from macroscopic and microscopic perspectives, by using a fifteen-day hydroponic experiments. The influence of magnesium (Mg) on Cu phytotoxicity to, and accumulation and translocation in grapevines was also observed. For phytotoxicity effect, results showed that a relative low median growth inhibition level of Cu was found for grapevine roots (0.809–3.671μM). Moreover, Cu toxicity was significantly alleviated by Mg treatment at Mg2+ activity between 0.15 and 2.01mM. For accumulation and translocation effects, results indicated that competition for binding sites between Cu and Mg occurred for roots; however, Mg and Cu levels in stems and leaves were not affected by solution metals concentration. At Cu concentration less than 1μM, the translocation of Cu was decreased significantly for the highest Mg treatment; at Cu concentrations greater than 5μM, no obvious change was observed in leaf TF value between Mg treatments, while an increasing trend of stem TF value was observed with increasing Mg. These results suggest that the toxic effect resulted from metals depend not only on the competition of coexistent cations for plasma membrane surface, but also on the transport and distribution of toxic metals in physiological active sites in plants.

Akaline, saline and mixed saline–alkaline stresses induce physiological and morpho‐anatomical changes in Lotus tenuis shoots

Saline, alkaline and mixed saline-alkaline conditions frequently co-occur in soil. In this work, we compared these plant stress sources on the legume Lotus tenuis, regarding their effects on shoot growth and leaf and stem anatomy. In addition, we aimed to gain insight on the plant physiological status of stressed plants. We performed pot experiments with four treatments: control without salt (pH = 5.8; EC = 1.2 dS·m(-1)) and three stress conditions, saline (100 mM NaCl, pH = 5.8; EC = 11.0 dS·m(-1)), alkaline (10 mM NaHCO3, pH = 8.0, EC = 1.9 dS·m(-1)) and mixed salt-alkaline (10 mM NaHCO3 + 100 mM NaCl, pH = 8.0, EC = 11.0 dS·m(-1)). Neutral and alkaline salts produced a similar level of growth inhibition on L. tenuis shoots, whereas their mixture exacerbated their detrimental effects. Our results showed that none of the analysed morpho-anatomical parameters categorically differentiated one stress from the other. However, NaCl- and NaHCO3 -derived stress could be discriminated to different extents and/or directions of changes in some of the anatomical traits. For example, alkalinity led to increased stomatal opening, unlike NaCl-treated plants, where a reduction in stomatal aperture was observed. Similarly, plants from the mixed saline-alkaline treatment characteristically lacked palisade mesophyll in their leaves. The stem cross-section and vessel areas, as well as the number of vascular bundles in the sectioned stem were reduced in all treatments. A rise in the number of vessel elements in the xylem was recorded in NaCl-treated plants, but not in those treated exclusively with NaHCO3.

NGAL Can Alternately Mediate Sunitinib Resistance in Renal Cell Carcinoma

Serum NGAL is highly expressed in patients with advanced renal cancer treated with sunitinib. We investigated the role of NGAL in sunitinib resistance in renal cell carcinoma to identify potential tactics to overcome it. NGAL expression was correlated with sunitinib sensitivity. Vascular endothelial growth factor related upstream Ras, Erk1/2 and STAT1 phosphorylation activity in Caki-1 and NGAL transfected Caki-1 cells after sunitinib treatment was analyzed using Western blot. NGAL and vascular endothelial growth factor-A interaction with sunitinib therapeutic efficacy was monitored in renal cell carcinoma tumor xenografted mice by tumor growth inhibition, serum NGAL and vascular endothelial growth factor-a levels, and microscopic examination of tumor microvascular density. Sunitinib cytotoxicity in various renal cell carcinoma cell lines was reversibly related to NGAL expression. Sunitinib showed the lowest 50% inhibitory concentration (5.53 μM) in Caki-1 cells, which had the lowest NGAL expression of these renal cell carcinoma cell lines. After sunitinib treatment adding NGAL inhibited Ras and Erk1/2 phosphorylation but activated STAT1α phosphorylation in Caki-1 cells and Caki-1 cells transfected with NGAL. In a xenograft mouse model sunitinib significantly inhibited tumor growth in Caki-1 mice. NGAL transfected Caki-1 mice had higher serum NGAL and lower vascular endothelial growth factor-A than Caki-1 mice. Microvascular density was decreased in Caki-1 mice with sunitinib treatment. NGAL in tumor cells may show crosstalk with vascular endothelial growth factor-a and alternative activation in stimulating tumor growth during sunitinib treatment. It may become a therapeutic target to reverse sunitinib resistance in renal cell carcinoma.

Novel Tuberculostatic Agents Suitable for Treatment of Mycobacterium tuberculosis Infections of the Central Nervous System

Aims:&nbsp;To demonstrate the efficacy of five small molecule compounds for inhibiting the growth of&nbsp;Mycobacterium tuberculosis.&nbsp;To present evidence that these compounds will penetrate into the central nervous system.Study Design:&nbsp;Five small molecule compounds bearing a hydrazide group were synthesized utilizing microwave excitation. These compounds were then placed into tissue culture with&nbsp;Mycobacterium tuberculosis&nbsp;at various concentrations for evaluation of bacterial growth inhibition.Place and Duration of Study:&nbsp;The compounds to be tested were prepared at the University of Nebraska Chemistry Department August 2013. The evaluation of antibacterial activity was determined at the Texas A&amp;M Health Science Center during October to December of 2013.Methodology:&nbsp;Applying microwave excitation for generation of hydrazide groups within the structure of small molecule carboxylic acids, five agents were prepared for evaluation of bacterial growth inhibition. These agents were dissolved into tissue culture media at various concentrations. Having various levels of tuberculostatic agents, then tuberculosis bacteria were added to determine level of growth inhibition. Growth inhibition of the bacteria was achieved and measured by compound concentration for comparison and evaluation.Results:&nbsp;Five compounds having a hydrazide functional group greatly inhibited the growth of&nbsp;Mycobacterium tuberculosis.&nbsp;All five agents had molecular weight less than 215 grams/mole and polar surface area of less than 70 Angstroms2. Values of Log P ranged from -0.226 to 0.998. Values of Log BB (Log [Cbrain/Cblood]) ranged from -0.711 to - 0.525, with a range in central nervous system penetration Cbrain/Cblood of 0.195 to 0.299. All compounds showed zero violations of the Rule of 5. Substantial inhibition of bacterial growth was observed at concentrations as low as 30 micrograms/mL, as measured by optical density and colony forming units.Conclusion:&nbsp;These five hydrazide compounds substantially decreased the proliferation of tuberculosis bacteria at concentrations as low as 30 micrograms/mL. In addition, their physicochemical properties are shown to allow high levels of penetration into the central nervous system.

Inhibition of the 26S proteasome as a possible mechanism for toxicity of heavy metal species

In this paper we report on the synthesis of five metal complexes coordinated to the [NN′O] ligand HLiodo (2,4-diiodo-6-((pyridine-2-ylmethylamino)methyl)phenol), namely [AlIII(Liodo)2]ClO4 (1), [CdII(Liodo)Cl]·H2O (2), [HgII(Liodo)2]·4DMSO (3), [PbII(Liodo)NO3] (4), and [SnIV(Liodo)Cl3] (5). Species 1–5 are thoroughly characterized by spectroscopic and spectrometric methods, as well as by elemental analysis. X-ray crystallography results for complex 3 indicate the presence of Hg(II) ion hexacoordinated to two facially oriented [NN′O] ligands, whereas for complex 5 an Sn(IV) ion chelates to one deprotonated ligand and three chlorido coligands. The toxicity of species 1–5 is tested against transformed human prostate epithelial cells CRL2221 and we observe that the five complexes demonstrate high levels of cell growth inhibition in a dose-dependent manner. In order to evaluate the relationship between these species and the proteasome, we test 1–5 against purified 20S, CRL2221 cell extracts, and intact cells, followed by the measurement of the percent chymotrypsin-like activity inhibition levels. Results suggest a good correlation between the toxicity of [HgII(Liodo)2]·4DMSO (3) and proteasome inhibition.

Exploiting the Synergy between Carboplatin and ABT-737 in the Treatment of Ovarian Carcinomas

Platinum drug-resistance in ovarian cancers mediated by anti-apoptotic proteins such as Bcl-xL is a major factor contributing to the chemotherapeutic resistance of recurrent disease. Consequently, concurrent inhibition of Bcl-xL in combination with chemotherapy may improve treatment outcomes for patients. Here, we develop a mathematical model to investigate the potential of combination therapy with ABT-737, a small molecule inhibitor of Bcl-xL, and carboplatin, a platinum-based drug, on a simulated tumor xenograft. The model is calibrated against in vivo experimental data, wherein xenografts established in mice were treated with ABT-737 and/or carboplatin on a fixed periodic schedule. The validated model is used to predict the minimum drug load that will achieve a predetermined level of tumor growth inhibition, thereby maximizing the synergy between the two drugs. Our simulations suggest that the infusion-duration of each carboplatin dose is a critical parameter, with an 8-hour infusion of carboplatin given weekly combined with a daily bolus dose of ABT-737 predicted to minimize residual disease. The potential of combination therapy to prevent or delay the onset of carboplatin-resistance is also investigated. When resistance is acquired as a result of aberrant DNA-damage repair in cells treated with carboplatin, drug delivery schedules that induce tumor remission with even low doses of combination therapy can be identified. Intrinsic resistance due to pre-existing cohorts of resistant cells precludes tumor regression, but dosing strategies that extend disease-free survival periods can still be identified. These results highlight the potential of our model to accelerate the development of novel therapeutics such as BH3 mimetics.

Pharmacokinetic–pharmacodynamic integration and modelling of florfenicol in calves

Florfenicol was administered subcutaneously to 10 calves at a dose of 40mg/kg. Pharmacokinetic-pharmacodynamic (PK-PD) integration and modelling of the data were undertaken using a tissue cage model, which allowed comparison of microbial growth inhibition profiles in three fluids, serum, exudate and transudate. Terminal half-lives were relatively long, so that florfenicol concentrations were well maintained in all three fluids. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration were determined in vitro for six strains each of the calf pneumonia pathogens, Mannhemia haemolytica and Pasteurella multocida. An PK-PD integration for three serum indices provided mean values for P.multocida and M.haemolytica, respectively, of 12.6 and 10.4 for Cmax /MIC, 183 and 152h for AUC0-24h /MIC and 78 and 76h for T>MIC. Average florfenicol concentrations in serum exceeded 4×MIC and 1.5×MIC for the periods 0-24 and 48-72h, respectively. Ex vivo growth inhibition curves for M.haemolytica and P.multocida demonstrated a rapid (with 8h of exposure) and marked (6 log10 reduction in bacterial count or greater) killing response, suggesting a concentration-dependent killing action. During 24-h incubation periods, inhibition of growth to a bacteriostatic level or greater was maintained in serum samples collected up to 96h and in transudate and exudate samples harvested up to 120h. Based on the sigmoidal Emax relationship, PK-PD modelling of the ex vivo time-kill data provided AUC0-24h /MIC serum values for three levels of growth inhibition, bacteriostatic, bactericidal and 4 log10 decrease in bacterial count; mean values were, respectively, 8.2, 26.6 and 39.0h for M.haemolytica and 7.6, 18.1 and 25.0h for P.multocida. Similar values were obtained for transudate and exudate. Based on pharmacokinetic and PK-PD modelled data obtained in this study and scientific literature values for MIC distributions, Monte Carlo simulations over 100000 trials were undertaken to predict once daily dosages of florfenicol required to provide 50% and 90% target attainment rates for three levels of growth inhibition, namely, bacteriostasis, bactericidal action and 4 log10 reduction in bacterial count.

Inhibition of ochratoxigenic moulds by Debaryomyces hansenii strains for biopreservation of dry-cured meat products

The ability of the osmotolerant yeast Debaryomyces hansenii to inhibit Penicillium nordicum, the most common ochratoxigenic mould encountered in dry-cured meat products, was evaluated. The antagonistic effect of ten D. hansenii strains isolated from dry-cured ham was screened in vitro using malt extract media and meat extract peptone media with the water activity (aw) adjusted to 0.97 and 0.90. A significant inhibition of the two tested P. nordicum strains by D. hansenii cells and cell-free supernatants was observed. At 0.97 aw, increasing D. hansenii inoculum concentrations significantly improved the inhibition of mould growth on solid medium, whereas at 0.90 aw this was not always the case. As observed by bright field microscopy, most D. hansenii strains were able to delay P. nordicum spore germination when co-cultured in malt extract broth. D. hansenii FHSCC 253H showed the highest overall in vitro inhibition of ochratoxigenic mould growth, and was therefore chosen for co-cultivation assays in dry-cured ham slices incubated at 0.94 and 0.84 aw simulating ham ripening. Regardless of the experimental conditions tested, lower levels of the inoculated P. nordicum strain were detected in co-cultivation batches compared with batches without D. hansenii. The highest level of mould growth inhibition was observed in batches at 0.94 aw. Ochratoxin A (OTA) production in ham samples was detected by HPLC-MS. Co-culturing of P. nordicum with D. hansenii FHSCC 253H resulted in lower OTA levels compared with control samples without D. hansenii. The decrease of the mycotoxin presence due to D. hansenii FHSCC 253H was more efficient at 0.94 aw (OTA was below the detection limit). In conclusion, D. hansenii is potentially suitable as a biopreservative agent for preventing ochratoxigenic mould growth and OTA accumulation in dry-cured meat products. The inoculation of D. hansenii should be made at the beginning of processing (at the end of post salting) when the aw of the product is still high (near 0.94). This action in addition to application of appropriate hygienic actions and control of temperature and relative humidity throughout ripening is required to reduce health risks due to OTA exposure.

TAT‐modified mixed micelles as biodegradable targeting and delivering system for cancer therapeutics

ABSTRACTA mixed micellar system of novel function was designed and synthesized by co‐assembling TAT (cell penetrating peptide)‐modified poly (ethylene glycol)‐poly(l‐lactide) (PEG‐PLA) copolymer with the drug‐conjugated poly(ethylene glycol)‐b‐poly(l‐lactide‐co‐2‐methyl‐2‐carboxyl‐propylene carbonate) (mPEG‐b‐P(LA‐co‐MCC)) copolymer. UV‐Vis, Matrix‐assisted laser desorption/ionization time‐of‐flight, and XPS were used to ensure the successful modification of the copolymers with TAT and anti‐tumor drugs. The size of spherical nanomicelles increased from 50 to 60 nm as of empty polymeric micelles to 100–150 nm as of drug‐loaded ones, determined by dynamic light scattering and TEM. Daunorubicin was selected as model drug for in vitro evaluations on different cell lines. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay clearly indicated an improved cell growth inhibition of the TAT‐modified mixed micelles. While green fluorescent protein was used as a marker for the mixed micelle, small amount of DMSO was necessary to enhance the accumulation of the mixed micelles in cell lines Caski. Mediated by TAT, mixed micelles containing Apoptin (a tumor‐specific apoptosis drug) showed higher level of tumor cell internalization and growth inhibition than that of mixed micelles without TAT modification. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 4598–4607, 2013

Blockade of NF-κB nuclear translocation results in the inhibition of the invasiveness of human gastric cancer cells

The aim of the present study was to investigate the effect of the nuclear factor-κB (NF-κB) p65 inhibitor, SN50, on the invasiveness and mechanisms of SGC7901 human gastric carcinoma cell xenografts in nude mice. Nude mice were randomly divided into model control and SN50 treatment groups. On days 5, 10 and 15 following treatment, the tumor samples were observed and a selection of parameters were recorded, including the level of tumor growth inhibition, the pathological changes in the tumor specimens, the expression levels of matrix metalloproteinase-9 (MMP-9), proliferating cell nuclear antigen (PCNA), tissue inhibitor of metalloproteinases type-1 (TIMP-1) and vascular endothelial growth factor (VEGF) and the apoptosis indices in the tumor samples. The results demonstrated that treating the tumor with SN50 for 5, 10 and 15 days inhibited carcinoma growth in comparison with the control group. Hematoxylin and eosin (HE) staining indicated that the level of inhibition increased progressively, in correlation with apoptosis. The expression of the MMP-9, PCNA and VEGF proteins was observed to be downregulated, while that of the TIMP-1 protein was shown to be upregulated, using immunohistochemical staining. In conclusion, the NF-κB p65 inhibitor, SN50, inhibited the invasiveness of the gastric cancer cells by downregulating the protein expression of MMP-9, PCNA and VEGF and upregulating the protein expression of TIMP-1. It was further suggested that SN50 may be a molecular target of anti-invasion therapy for gastric cancer, and that the inhibition of the NF-κB p65 signaling pathway may be considered as a potential strategy for treating gastric cancer.

Inhibition of Wee1, AKT, and CDK4 Underlies the Efficacy of the HSP90 Inhibitor XL888 in an In Vivo Model of NRAS-Mutant Melanoma

The HSP90 inhibitor XL888 is effective at reversing BRAF inhibitor resistance in melanoma, including that mediated through acquired NRAS mutations. The present study has investigated the mechanism of action of XL888 in NRAS-mutant melanoma. Treatment of NRAS-mutant melanoma cell lines with XL888 led to an inhibition of growth, G2-M phase cell-cycle arrest, and the inhibition of cell survival in three-dimensional spheroid and colony formation assays. In vitro, HSP90 inhibition led to the degradation of ARAF, CRAF, Wee1, Chk1, and cdc2 and was associated with decreased mitogen-activated protein kinase (MAPK), AKT, mTOR, and c-jun NH2 kinase (JNK) signaling. Apoptosis induction was associated with increased BIM expression and a decrease in the expression of the prosurvival protein Mcl-1. The critical role of increased BIM and decreased Mcl-1 expression in the survival of NRAS-mutant melanoma cell lines was shown through siRNA knockdown and overexpression studies. In an animal xenograft model of NRAS-mutant melanoma, XL888 treatment led to reduced tumor growth and apoptosis induction. Important differences in the pattern of client degradation were noted between the in vivo and in vitro studies. In vivo, XL888 treatment led to degradation of CDK4 and Wee1 and the inhibition of AKT/S6 signaling with little or no effect observed upon ARAF, CRAF, or MAPK. Blockade of Wee1, using either siRNA knockdown or the inhibitor MK1775, was associated with significant levels of growth inhibition and apoptosis induction. Together, these studies have identified Wee1 as a key target of XL888, suggesting novel therapeutic strategies for NRAS-mutant melanoma.

Impact of Polymers on Crystal Growth Rate of Structurally Diverse Compounds from Aqueous Solution

The presence of an effective crystal growth inhibitor in solution is desirable to prolong supersaturation since residual crystalline material in an amorphous formulation resulting from the manufacturing process or formed during storage or dissolution can potentially have a significant impact on the extent and duration of supersaturation. In this study, the effectiveness of a group of chemically diverse polymers, including several recently synthesized cellulose derivatives, on solution crystal growth of three structurally diverse compounds (celecoxib, efavirenz, and ritonavir) was quantified at different extents of supersaturation and compared. Despite the different chemical properties and structures of the model compounds, nonspecific hydrophobic drug-polymer interactions appeared to be important in determining the impact of a given polymer on crystal growth for of all these drug compounds. Specific intermolecular interactions were also found to be important for crystal growth inhibition of celecoxib and efavirenz by the hydrophilic polymer, PVPVA. These interactive forces-hydrophobicity and specific intermolecular interactions-are likely to promote adsorption of the polymer onto the surface of the crystalline drugs, thus influencing crystal growth. The effectiveness of the polymers also depended on the rate of crystallization of the drug molecules. At a similar supersaturation ratio of ∼1.2, ritonavir and celecoxib had slower normalized crystal growth rates (0.20 and 0.91 mg min(-1) m(-2), respectively), while the normalized crystal growth rate of efavirenz was significantly higher (2.97 mg min(-1) m(-2)), resulting in lower levels of crystal growth inhibition by the polymers for efavirenz.

Na+ conductivity of the Na+-driven flagellar motor complex composed of unplugged wild-type or mutant PomB with PomA

PomA and PomB form the stator complex, which functions as a Na(+) channel, in the Na(+)-driven flagellar motor of Vibrio alginolyticus. The plug region of PomB is thought to regulate the Na(+) flow and to suppress massive ion influx through the stator channel. In this study, in order to measure the Na(+) conductivity of the unplugged stator, we over-produced a plug-deleted stator of the Na(+)-driven flagellar motor in Escherichia coli. The over-production of the plug-deleted stator in E. coli cells caused more severe growth inhibition than in Vibrio cells and that growth inhibition depended on the Na(+) concentration in the growth medium. Measurement of intracellular Na(+) concentration by flame photometry and fluorescent analysis with a Na(+) indicator, Sodium Green, revealed that over-production of the plug-deleted stator increased the Na(+) concentration in cell. Some mutations in the channel region of PomB or in the cytoplasmic region of PomA suppressed both the growth inhibition and the increase in intracellular Na(+) concentration. These results suggest that the level of growth inhibition correlates with the intracellular Na(+) concentration, probably due to the Na(+) conductivity through the stator due to the mutations.