A novel A33 promoter-based conditionally replicative adenovirus suppresses tumor growth and eradicates hepatic metastases in human colon cancer models

Purpose: A33 antigen is a membrane-bound protein expressed in intestinal epithelium that is overexpressed in 95% of primary and metastatic colorectal carcinomas but is absent in most epithelial tissues and tumor types. We hypothesized that A33 promoter might be useful in the design of a conditionally replicative adenovirus for the treatment of colorectal cancer (CRC). Experimental Design: We cloned an A33 promoter fragment (A33Pr) that extends from -105 to +307 bp. Using luciferase activity as a reporter gene, we showed that A33Pr was active inCRC cell lines. We next constructed a conditionally replicative adenovirus named AV22EL where E1A was placed under the control of A33Pr. The tumor-specific oncolytic effect of AV22EL was investigated both in vitro and in vivo. Results: AV22EL induced specific in vitro lysis of human CRC cell lines that expressed A33 and have negligible lytic capacity on cells that lacked or hadminimal A33 expression, including normal human colonic cells. In vivo, a marked reduction of tumor growth and increased long-term survival rates were observed in nude mice xenografted with s.c. CRC tumors. Combination with 5-fluorouracil induced an additive effect in vitro with no toxic effects in vivo. Remarkably, AV22EL completely eliminated established hepatic metastases in >90% of mice and restored hepatic function according to biochemical parameters. Its systemic administration induced E1A expression only in the hepatic metastasis but not in normal organs. Conclusions: These data show that AV22EL is a stringently regulated and potent oncolytic agent for the treatment of CRC.Facultad de Ciencias Veterinaria

Expression of a suicidal gene under control of the human secreted protein acidic and rich in cysteine (SPARC) promoter in tumor or stromal cells led to the inhibition of tumor cell growth

The successful use of transcriptional targeting for cancer therapy depends on the activity of a given promoter inside the malignant cell. Because solid human tumors evolve as a "cross-talk" between the different cell types within the tumor, we hypothesized that targeting the entire tumor mass might have better therapeutic effect. Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein overexpressed in different human cancers malignant melanomas both in the malignant cells compartment as in the stromal one (fibroblasts and endothelial cells). We have shown that expression of the herpes simplex virus - thymidine kinase (TK) gene driven by the SPARC promoter in combination with ganciclovir inhibited human melanoma cell growth in monolayer as well as in multicellular spheroids. This inhibitory effect was observed both in homotypic spheroids composed of melanoma cells alone as well as in spheroids made of melanoma cells and stromal cells. Expression of the TK gene was also efficient to inhibit the in vivo tumor growth of established melanomas when TK was expressed either by the malignant cells. Our data suggest that the use of therapeutic genes driven by SPARC promoter could be a valuable strategy for cancer therapy aiming to target all the cellular components of the tumor mass.Facultad de Ciencias Veterinaria

Expression of a suicidal gene under control of the human secreted protein acidic and rich in cysteine (SPARC) promoter in tumor or stromal cells led to the inhibition of tumor cell growth

The successful use of transcriptional targeting for cancer therapy depends on the activity of a given promoter inside the malignant cell. Because solid human tumors evolve as a "cross-talk" between the different cell types within the tumor, we hypothesized that targeting the entire tumor mass might have better therapeutic effect. Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein overexpressed in different human cancers malignant melanomas both in the malignant cells compartment as in the stromal one (fibroblasts and endothelial cells). We have shown that expression of the herpes simplex virus - thymidine kinase (TK) gene driven by the SPARC promoter in combination with ganciclovir inhibited human melanoma cell growth in monolayer as well as in multicellular spheroids. This inhibitory effect was observed both in homotypic spheroids composed of melanoma cells alone as well as in spheroids made of melanoma cells and stromal cells. Expression of the TK gene was also efficient to inhibit the in vivo tumor growth of established melanomas when TK was expressed either by the malignant cells. Our data suggest that the use of therapeutic genes driven by SPARC promoter could be a valuable strategy for cancer therapy aiming to target all the cellular components of the tumor mass.Facultad de Ciencias Veterinaria

Tumor Associated Stromal Cells Play a Critical Role on the Outcome of the Oncolytic Efficacy of Conditionally Replicative Adenoviruses

The clinical efficacy of conditionally replicative oncolytic adenoviruses (CRAd) is still limited by the inefficient infection of the tumor mass. Since tumor growth is essentially the result of a continuous cross-talk between malignant and tumor-associated stromal cells, targeting both cell compartments may profoundly influence viral efficacy. Therefore, we developed SPARC promoter-based CRAds since the SPARC gene is expressed both in malignant cells and in tumor-associated stromal cells. These CRAds, expressing or not the Herpes Simplex thymidine kinase gene (Ad-F512 and Ad(I)-F512-TK, respectively) exerted a lytic effect on a panel of human melanoma cells expressing SPARC; but they were completely attenuated in normal cells of different origins, including fresh melanocytes, regardless of whether cells expressed or not SPARC. Interestingly, both CRAds displayed cytotoxic activity on SPARC positive-transformed human microendothelial HMEC-1 cells and WI-38 fetal fibroblasts. Both CRAds were therapeutically effective on SPARC positive-human melanoma tumors growing in nude mice but exhibited restricted efficacy in the presence of co-administered HMEC-1 or WI-38 cells. Conversely, co-administration of HMEC-1 cells enhanced the oncolytic efficacy of Ad(I)-F512-TK on SPARC-negative MIA PaCa-2 pancreatic cancer cells in vivo. Moreover, conditioned media produced by stromal cells pre-infected with the CRAds enhanced the in vitro viral oncolytic activity on pancreatic cancer cells, but not on melanoma cells. The whole data indicate that stromal cells might play an important role on the outcome of the oncolytic efficacy of conditionally replicative adenoviruses.Facultad de Ciencias Veterinaria

Universal structure of the edge states of the fractional quantum Hall states

We present an effective theory for the bulk fractional quantum Hall states on the Jain sequences on closed surfaces and show that it has a universal form whose structure does not change from fraction to fraction. The structure of this effective theory follows from the condition of global consistency of the flux attachment transformation on closed surfaces. We derive the theory of the edge states on a disk that follows naturally from this globally consistent theory on a torus. We find that, for a fully polarized two-dimensional electron gas, the edge states for all the Jain filling fractions $\nu=p/(2np+1)$ have only one propagating edge field that carries both energy and charge, and two non-propagating edge fields of topological origin that are responsible for the statistics of the excitations. Explicit results are derived for the electron and quasiparticle operators and for their propagators at the edge. We show that these operators create states with the correct charge and statistics. It is found that the tunneling density of states for all the Jain states scales with frequency as $|\omega|^{(1-\nu)/\nu}$.Comment: 10 page

Gauge Invariance and Finite Temperature Effective Actions of Chern-Simons Gauge Theories with Fermions

We discuss the behavior of theories of fermions coupled to Chern-Simons gauge fields with a non-abelian gauge group in three dimensions and at finite temperature. Using non-perturbative arguments and gauge invariance, and in contradiction with perturbative results, we show that the coefficient of the Chern-Simons term of the effective actions for the gauge fields at finite temperature can be {\it at most} an integer function of the temperature. This is in a sense a generalized no-renormalization theorem. We also discuss the case of abelian theories and give indications that a similar condition should hold there too. We discuss consequences of our results to the thermodynamics of anyon superfluids and fractional quantum Hall systems.Comment: Revtex, multico

Fermionic Chern-Simons theory for the Fractional Quantum Hall Effect in Bilayers

We generalize the fermion Chern-Simons theory for the Fractional Hall Effect (FQHE) which we developed before, to the case of bilayer systems. We study the complete dynamic response of these systems and predict the experimentally accessible optical properties. In general, for the so called $(m, m, n)$ states, we find that the spectrum of collective excitations has a gap, and the wave function has the Jastrow-Slater form, with the exponents determined by the coefficients $m$, and $n$. We also find that the $(m,m,m)$ states, {\it i.~e.~}, those states whose filling fraction is $1\over m$, have a gapless mode which may be related with the spontaneous appearance of the interlayer coherence. Our results also indicate that the gapless mode makes a contribution to the wave function of the $(m,m,m)$ states analogous to the phonon contribution to the wave function of superfluid $\rm{He}_4$. We calculate the Hall conductance, and the charge and statistics of the quasiparticles. We also present an $SU(2)$ generalization of this theory relevant to spin unpolarized or partially polarized single layers.Comment: 55 pages, Urbana Prepin

Spatio-temporal dynamics of dengue 2009 outbreak in Córdoba city, Argentina.

During 2009 the biggest dengue epidemic to date occurred in Argentina, affecting almost half the country. We studied the spatio-temporal dynamics of the outbreak in the second most populated city of the country, Córdoba city. Confirmed cases and the results of an Aedes aegypti monitoring during the outbreak were geolocated. The imported cases began in January, and the autochthonous in March. Thirty-three percent of the 130 confirmed cases were imported, and occurred mainly at the center of the city. The autochthonous cases were more frequent in the outskirts, specially in the NE and SE. Aedes aegypti infestation showed no difference between neighborhoods with or without autochthonous cases, neither between neighborhoods with autochthonous vs. imported cases. The neighborhoods with imported cases presented higher population densities. The majority of autochthonous cases occurred at ages between 25 and 44 years old. Cases formed a spatio-temporal cluster of up to 20 days and 12 km. According to a athematical model that estimates the required number of days needed for transmission according to daily temperature, the number of cases begun to fall when more than 15.5 days were needed. This may be a coarse estimation of mean mosquito survival in the area, provided that the study area is close to the global distribution limit of the vector, and that cases prevalence was very low.Fil: Estallo, Elizabet Lilia. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones Entomológicas de Córdoba; ArgentinaFil: Carbajo, Anibal Eduardo. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Grech, Marta Gladys. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Esquel. Departamento de Biología. Laboratorio de Investigaciones en Ecología y Sistemática Animal; ArgentinaFil: Frias Cespedes, M.. Ministerio de Salud de la Provincia de Córdoba; ArgentinaFil: Lopez, L.. Ministerio de Salud de la Provincia de Córdoba; ArgentinaFil: Lanfri, M. A.. Comision Nacional de Actividades Espaciales; ArgentinaFil: Ludueña Almeida, Francisco F.. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones Entomológicas de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Almiron, Walter Ricardo. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones Entomológicas de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas; Argentin

Co-expression of inducible nitric oxide synthase and arginases in different human monocyte subsets. Apoptosis regulated by endogenous NO

Human monocyte subsets, isolated from cultures of mononuclear cells, or freshly obtained from patients with multiple sclerosis, Graves' disease or pemphigus vulgaris, differed in phenotype, apoptotic features, mRNA levels of arginase II (A-II) and the inducible form of nitric oxide synthase (iNOS). Liver-type arginase I mRNA was present in all subsets. Apoptosis was followed by the expression of T cell intracellular antigen (TIA) and the simultaneous detection of DNA stainability by propidium iodine and annexin V binding. Apoptosis was practically absent both in activated CD14(++)CD33(++)DR(++)CD25(++)CD69(++)CD71(++/+) CD16(-) cells, expressing A-II mRNA and having arginase activity, but not iNOS mRNA, and in not fully mature large CD14(++)CD16(+)CD23(+)DR(++) monocytes, expressing simultaneously both mRNAs and having both enzyme activities. However, differentiated small CD14(+/++)CD16(+)CD69(+)CD25(+/-)CD71(++)CD23(+) DR(++) monocytes, expressing high levels of iNOS mRNA, exhibited apoptotic signs. Amounts of NO synthesised by monocytes co-expressing iNOS and arginase changed with the addition of arginine or an iNOS inhibitor; in that case a correlation of NO production and apoptotic features was observed. Data suggest a regulatory role for endogenous NO in apoptosis of stimulated and differentiated monocytes, and also that iNOS and A-II, when simultaneously present, could control the production of NO as a consequence of their competition for arginine