José Mariano Ruiz de Almodóvar

University of Granada | UGR · Centro de Investigación Biomédica (CIBM)

Peroxidase-mimicking activity of manganese ferrite nanoparticles was studied, based on the oxidation of TMB (3,3’,5,5’-tetramethylbenzidine) by H2O2 at pH 4 using acetate and citrate buffers. The aim of this study was to examine this reaction not only by enzymology (Michaelis-Menten kinetics model) but also by surface science methods of heterogeneo...

We have previously shown that the combination of radiotherapy with human umbilical-cord-derived mesenchymal stromal/stem cells (MSCs) cell therapy significantly reduces the size of the xenotumors in mice, both in the directly irradiated tumor and in the distant nonirradiated tumor or its metastasis. We have also shown that exosomes secreted from MS...

Previously we have shown that the combination of radiotherapy with human-umbilical-cord-derived mesenchymal stem-cell therapy significantly reduces the size of the xenotumours in mice, both in the directly irradiated tumour and in the distant non-irradiated tumour or in its metastasis. We have also shown that exosomes secreted from mesenchymal stem...

In this paper, we summarize published articles and experiences related to the attempt to improve radiotherapy outcomes and, thus, to personalize the radiation treatment according to the individual characteristics of each patient. The evolution of ideas and the study of successively published data have led us to envisage new biophysical models for t...

Background: We have recently shown that radiotherapy may not only be a successful local and regional treatment but, when combined with MSCs, may also be a novel systemic cancer therapy. This study aimed to investigate the role of exosomes derived from irradiated MSCs in the delay of tumor growth and metastasis after treatment with MSC + radiothera...

The outcome of radiotherapy treatment might be further improved by a better understanding of individual variations in tumor radiosensitivity and normal tissue reactions, including the bystander effect. For many tumors, however, a definitive cure cannot be achieved, despite the availablity of more and more effective cancer treatments Therefore, any...

Poly (ADP-ribose) polymerase (PARP) inhibitors are particularly efficient against tumors with defects in the homologous recombination repair pathway. Nonetheless poly(ADP-ribosylation) (PARylation) modulates prometastasic activities and adaptation of tumor to a hostile microenvironment. Modulation of metastasis-promoting traits is possible through...

Glioblastoma multiforme (GBM) is the most common primary brain tumour in adults and one of the most aggressive cancers. PARP-1 is a nuclear protein involved in multiple facets of DNA repair and transcriptional regulation. In this study we dissected the action of PARP inhibition in different GBM cell lines with either functional or mutated PTEN that...

Tumor expression of estrogen receptor (ER) is an important marker of prognosis, and is predictive of response to endocrine therapy in breast cancer. Several studies have observed that epigenetic events, such methylation of cytosines and deacetylation of histones, are involved in the complex mechanisms that regulate promoter transcription. However,...

We have investigated the capacity of activated carbon cloth to support the growth and differentiation of human mesenchymal umbilical-cord stromal stem cells. Our results demonstrate that this scaffold provides suitable conditions for the development of cell-derived matrix proteins and facilitates the growth of undifferentiated stem cells with the a...

2D-DIGE (Differential In-Gel Electrophoresis). HUVEC were solubilized in 2D-DIGE sample buffer (40 mM Tris, 7 M Urea, 2 M Thiourea, 1% ASB-14), sonicated and then the concentration was determined using the RC hDC Protein Assay (Bio- Rad). Fifty µg of protein was then labelled with 400 pmol of CyDye DIGE Fluor minimal dyes (GE Healthcare) and incuba...

PARP inhibitors decrease VEGF-induced tube formation in HUVECs in vitro and in vivo. Cells were collected and seeded in Matrigel-coated 48-well plates and then incubated in the absence (Control) or presence of VEGF and DPQ (40 µM) or PJ34 (20 µM). After 48 h, the morphological changes of the cells and any tubes formed were observed and recorded und...

Western-blot (A) and immunofluorescence (B) of PARP activity inhibition in G361cells treated with the PARP-1 inhibitor, DPQ. (TIF)

Treatment with the PARP inhibitor olaparib decreased metastatic spread of melanoma cells. C57BL/6 mice were inoculated with B16-F10-luc cells a treated with the PARP inhibitor olaparib (50 mg/kg) as explained in Methods. Results obtained on the 17th day are shown for quantitation. *P<0.05 olaparib versus control using the Mann-Whitney u-test. (TIF)

Cell proliferation and apoptosis are not affected by PARP inhibition in metastasis. Mitosis and apoptosis were evaluated in histological metastasis slides using morphological criteria and evaluated in a blind observation by two different pathologists. (TIF)

Quantitation of western-blots. All western blots shown have been quantified using Quantity One software from Bio-Rad. (PDF)

Development of subcutaneous xenografts of melanoma is reduced by treatment with the PARP inhibitor DPQ. (A) C57BL/6 albino mice (Jackson Laboratories, Philadelphia, USA), were inoculated with B16-F10-luc cells as explained in Methods. Localization and intensity of luciferase expression was monitored by in vivo bioluminescence imaging. Quantitation...

Results for all the different markers are presented. Samples were analyzed in a blind fashion by two different pathologists. (TIF)

PARP inhibition reduced the expression of Vimentin and Snail1 and up-regulates E-cadherin murine melanoma cells. Cells were treated with either of the PARP inhibitors DPQ (40 µM) (not shown), PJ34 at 10 µM or KU0058948 (100 nM) during 22 hours. IF (A), western-blot (B) or qPCR (C) were performed to evaluate the impact of PARP inhibition on EMT mark...

IHC evaluation of PARP-1 and EMT markers in human melanoma tissue array. Expression of PARP-1 and EMT markers in nodular and metastatic human melanoma. PARP-1 expression correlates with vimentin in nodular and metastatic melanoma. Snail1 and E-cadherin expression do not correlate with PARP-1 positivity. (TIF)

PARP inhibition can induce anti-neoplastic effects when used as monotherapy or in combination with chemo- or radiotherapy in various tumor settings; however, the basis for the anti-metastasic activities resulting from PARP inhibition remains unknown. PARP inhibitors may also act as modulators of tumor angiogenesis. Proteomic analysis of endothelial...

Background In cervical postoperative radiotherapy, the target volume is usually the same as the extension of the previous dissection. We evaluated a protocol of selective irradiation according to the risk estimated for each dissected lymph node level. Methods Eighty patients with oral/oropharyngeal cancer were included in this prospective clinical...

ESTRO 31 S607 geneticin, stably transfected cells were obtained. To test stress-induced regulation of the CMV promoter, cells were exposed to IR (6 Gy) and CDDP (3microg/ml). Expression of reporter gene was determined by fluorescence microplate reader and fluorescence microscopy at day 3 post-treatment. Solid TS/A tumors were induced by subcutaneou...

In response to nutrient stress, cells start an autophagy program that can lead to adaptation or death. The mechanisms underlying the signaling from starvation to the initiation of autophagy are not fully understood. In the current study we show that the absence or inactivation of PARP-1 strongly delays starvation-induced autophagy. We have found th...

To examine direct and bystander radiation-induced effects in normal umbilical-cord stromal stem cell (HCSSC) lines and in human cancer cells. The UCSSC lines used in this study were obtained in our laboratory. Two cell lines (UCSSC 35 and UCSSC 37) and two human melanoma skin-cancer cells (A375 and G361) were exposed to ionizing radiation to measur...

Purpose: Posttranslational modifications are known to be altered in cancer cells, and loss of selected histone acetylation and methylation marks has recently been shown to predict patient outcome in human carcinoma. Materials: Since January 2002 to June 2005, 107 women with breast cancer and 108 control subjects were recruited. A sensitive SYBR gre...

DNA-damage assays, quantifying the initial number of DNA double-strand breaks induced by radiation, have been proposed as a predictive test for radiation-induced toxicity. Determination of radiation-induced apoptosis in peripheral blood lymphocytes by flow cytometry analysis has also been proposed as an approach for predicting normal tissue respons...

Numerous hypermethylated genes have been reported in breast cancer, and the silencing of these genes plays an important role in carcinogenesis, tumor progression and diagnosis. These hypermethylated promoters are very rarely found in normal breast. It has been suggested that aberrant hypermethylation may be useful as a biomarker, with implications...

e13648 Background: To determine whether estrogen receptor (ER) (+) and ER (-) status relates to epigenetic changes in breast cancer-related genes and to correlate with molecular breast cancer subtypes. Methods: From January 2002 to June 2005, we quantified methylation levels ERS1 gene in serum of 92 pts breast cancer. A PCR quantitative technique w...

e13619 Background: The CpG islands of many genes, which are usually unmethylated in normal tissue, are methylated in cancer. The aim of the study was to investigate the association between gene hypermethylation and the major clinicopathological features of breast cancer, diagnosis, and treatment response. Methods: We quantified methylation levels o...

Background: The CpG island methylator phenotype is associated with distinct clinicophatological characteristics as Estrogen Receptor (ESR1) positive and amplification of HER2 in breast cancer.Objetive: To evaluate epigenetics differences in tumor-related genes to ERS1 and HER2/neu status in primary breast cancer.Material and Methods: We quantified...

Activated carbon cloth was shown to be a good support for the growth and differentiation of mesenchymal stem cells (MSCs). By using this approach, we improved the cell culture procedure and reduced the laboratory time required to obtain a very large number of MSCs suitable for utilization or differentiation.

Poly(ADP-ribose) polymerases (PARPs) are defined as cell signaling enzymes that catalyze the transfer of ADP-ribose units from NAD+ to a number of acceptor proteins. PARP-1, the best characterized member of the PARP family, which currently comprises 18 members, is an abundant nuclear enzyme implicated in cellular responses to DNA injury provoked by...

Autophagy is a lysosome-dependent degradative pathway frequently activated in tumor cells treated with chemotherapy or radiation. PARP-1 has been implicated in different pathways leading to cell death and its inhibition potentiates chemotherapy-induced cell death. Whether PARP-1 participates in the cell's decision to commit to autophagy following D...

Resumen Lo que más preocupa a los profesionales de la medicina son los resultados de los procedimientos diagnósticos y terapéuticos que empleamos y proponemos. Nos gustaría que el diagnóstico fuese perfecto; esto es: que el método utilizado para diagnosticar la enfermedad fuese absolutamente fiable, sensible y específico y que la terapia fuese segu...

The aim of the present study was to investigate the association between gene hypermethylation and main clinicopathological features of breast cancer, including diagnosis and treatment response. A sensitive SYBR green methylation-specific PCR technique was used to analyze the utility of circulating DNA with CpG island hypermethylation of ESR1, APC,...

This 14-year-long study makes a novel contribution to the debate on the relationship between the in vitro radiosensitivity of peripheral blood lymphocytes and normal tissue reactions after radiation therapy. The aims were (1) to prospectively assess the degree and time of onset of skin side effects in 40 prospectively recruited consecutive patients...

Post-translational modification of proteins by poly(ADP-ribosyl)ation is involved in the regulation of a number of biological functions. While an 18 member superfamily of poly(ADP-ribose) polymerases (PARP)s has been described PARP-1 accounts for more than 90% of the poly(ADP-ribosyl)ating capacity of the cells. PARP-1 act as a DNA nick sensor and...

ATM and PARP-1 are two of the most important players in the cell's response to DNA damage. PARP-1 and ATM recognize and bound to both single and double strand DNA breaks in response to different triggers. Here we report that ATM and PARP-1 form a molecular complex in vivo in undamaged cells and this association increases after gamma-irradiation. AT...

Poly(ADP-ribose) polymerase (PARP)-1, an enzyme that catalyzes the attachment of ADP ribose to target proteins, acts as a component of enhancer/promoter regulatory complexes. In the present study, we show that pharmacologic inhibition of PARP-1 with 3,4-dihydro-5-[4-(1-piperidinyl)butoxyl]-1(2H)-isoquinolinone (DPQ) results in a strong delay in tum...

Fanconi anemia is a genetically heterogeneous recessive disease characterized mainly by bone marrow failure and cancer predisposition. Although it is accepted that Fanconi cells are highly sensitive to DNA crosslinking agents, their response to ionizing radiation is still unclear. Using pulsed-field gel electrophoresis, we have observed that radiat...

3-nitrotyrosine (NO2-Tyr) is thought to be a specific marker of cell injury during oxidative damage. We have evaluated the role of poly(ADP-ribose)polymerase-1 (PARP-1) in protein nitration after treatment of immortalized fibroblasts parp-1+/+ and parp-1-/- with the alkylating agent 2'-methyl-2'-nitroso-urea (MNU). Both cell lines showed increased...

p53 deficiency confers resistance to doxo (doxorubicin), a clinically active and widely used antitumour anthracycline antibiotic. The purpose of the present study was to investigate the reversal mechanism of doxo resistance by the potent PARP [poly(ADP-ribose) polymerase] inhibitor ANI (4-amino-1,8-naphthalimide) in the p53-deficient breast cancer...

Radiotherapy outcomes might be further improved by a greater understanding of the individual variations in normal tissue reactions that determine tolerance. Most published studies on radiation toxicity have been performed retrospectively. Our prospective study was launched in 1996 to measure the in vitro radiosensitivity of peripheral blood lymphoc...

Poly (ADP-ribose) polymerase-1 (PARP-1)-deficient mice are protected against septic shock, type I diabetes, stroke and inflammation. It is now accepted that inflammation and related events, such as activation of NF-kappaB, are key components in the initiation and progression of epithelial cancer and in particular in the neoplastic transformation of...

In this work we estimate the therapeutic gain that could be obtained using a radiotherapy programme in which doses were based on a radiosensitivity test that was able to predict the final response of normal tissues to radiation for each particular patient. To date, no good radiosensitivity assay has been demonstrated and by way of example we use an...

This study was undertaken to investigate whether hypermethylation in p16(INK4a) gene promoter could serve as plasma biomarker of bladder cancer. We examined the p16(INK4a) status using methylation-specific PCR in 86 cancer patients and 49 controls (31 healthy people and 18 patients with benign urological diseases). The p16(INK4a) methylation was fo...

Whenever radiation therapy is given with curative intent there is the risk of serious damage to normal tissue. This risk increases with the dose of radiation, as does the probability of local tumour control. In the attempt to cure, the doses reach a level that inevitably causes some undesirable adverse effects, ranging from undetectable, or minimal...

Reporting of the outcome of radiotherapy is not satisfactory without a description of the treatment-related side effects. The purposes of this paper were: (1) to evaluate the frequency and the severity of collateral skin reactions in a group of breast cancer patients; (2) to report the acute reactions using some current scoring systems and to compa...

The purpose of this study was to determine whether the distribution of sensitivities in breast cancer patients, measured using a DNA damage assay on lymphocytes, is likely to provide sufficient discrimination to enable the reliable identification of patients with abnormal sensitivities. Radiosensitivity (x) was assessed in 226 samples of lymphocyte...

The tumour suppressor protein p53 plays a key role in the cell's decision to arrest the cell cycle or undergo apoptosis following a genotoxic insult. p53 is stabilized and activated after DNA damage, however the cascade of events signalling from DNA lesions to p53 stabilization and activation is still controversial. Poly (ADP-ribosylation) of diffe...

We have investigated whether the protective role of the G2 checkpoint has increasing importance when the p53-dependent G1 checkpoint is inactivated. We have studied the differential effect of caffeine by clonogenic assays and flow cytometry in three human tumour cell lines with different functionality of p53 protein. The radiosensitizing effect of...

Apoptosis is a type of cellular death but also directly regulates tumorigenesis through different gene expression. This phenomenon is often used as end-point in studies of radio- and chemosensitivity of cancer cells. Restriction DNA fragments have been separated quickly, efficiently and successfully by capillary gel electrophoresis (CGE). In this s...

Apoptosis and necrosis are two different forms of cell death that can be induced by cytotoxic stress, such as ionizing radiation. We have studied the importance of apoptotic death induced after treatment with 6 Gy of gamma-irradiation in a panel of eight human tumour cell lines of different radiosensitivities. Three different techniques based on th...

The success of radiotherapy in eradicating tumours depends on the total radiation dose, but what limits this dose is the tolerance of the normal tissues within the treatment volume. Studies involving fibroblast survival have demonstrated the theoretical feasibility of a predictive assay of radiation sensitivity, but such an assay is still far from...

The success of radiotherapy in eradicating tumours depends on the total radiation dose, but what limits this dose is the tolerance of the normal tissues within the treatment volume. Studies involving fibroblast survival have demonstrated the theoretical feasibility of a predictive assay of radiation sensitivity, but such an assay is still far from...