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Rafael Molina

Publications and source records attributed to Rafael Molina.

At least 19 recordsLinked to original sources

Blind deconvolution using a variational approach to parameter, image, and blur estimation.

Following the hierarchical Bayesian framework for blind deconvolution problems, in this paper, we propose the use of simultaneous autoregressions as prior distributions for both the image and blur, and gamma distributions for the unknown parameters (hyperparameters) of the priors and the image formation noise. We show how the gamma distributions on the unknown hyperparameters can be used to prevent the proposed blind deconvolution method from converging to undesirable image and blur estimates and also how these distributions can be inferred in realistic situations. We apply variational methods to approximate the posterior probability of the unknown image, blur, and hyperparameters and propose two different approximations of the posterior distribution. One of these approximations coincides with a classical blind deconvolution method. The proposed algorithms are tested experimentally and compared with existing blind deconvolution methods.

Algorithms↗

Simultaneous genotyping of CYP2C9*2, *3, and 5' flanking region (C-1189T) polymorphisms in a Spanish population through a new minisequencing multiplex single-base extension analysis.

OBJECTIVE: To conduct a descriptive study on the prevalence of relevant cytochrome P450 2C9 (CYP2C9) polymorphisms--the *2, *3, and 5' flanking region (C-1189T)--in a Spanish population using a new minisequencing fluorescent method through a multiplex single base extension (SBE) analysis. METHOD: The method simultaneously and accurately genotypes the CYP2C9 polymorphisms studied and is available as a commercial protocol (SNaPshot). Various strategies, including restriction fragment length polymorphism (RFLP) and Taqman, were used to validate the methodology. RESULTS: The frequencies of alleles CYP2C9*2 (12%) and *3 (6.2%) were similar to those described for other Caucasian populations. The frequency of allele t at the 5' flanking region was 62%, which is close to the percentage reported in Japanese and French populations. The four haplotypes inferred in our samples and their frequencies were consistent with those reported in other studies. CONCLUSION: Our results confirm previously reported Caucasian frequencies for the CYP2C9*2 and *3 alleles and, for the first time, provide data on the frequency of the CYP2C9 5' flanking region (C-1189T), a recently described polymorphism, in a Spanish population. The SBE technique detects unequivocally the three polymorphisms in a single reaction, which makes it suitable for the analysis of CYP2C9 in the many therapeutic situations in which it is involved.

5' Flanking Region↗

Determination of microcystins in fish by solvent extraction and liquid chromatography.

A liquid chromatography electrospray mass spectrometry (LC/ESI/MS) method has been developed to identify and quantify microcystins in fish liver and intestine. Microcystins (MCs) were extracted from 500 mg sample with methanol-water (85:25, v/v) and the extracts concentrated to 250 microl. The parameters were optimized by a full factorial 2(3) design. Neither laborious pre-treatment nor clean up were necessary. MCs were separated using conventional C18 column and an acetonitrile-acidified water (pH 3) gradient. Negative samples (without MCs) were discriminated by liquid chromatography diode array detection (LC/DAD). The limits of detection (LOD) and the limits of quantification (LOQ) resulted equal for MC-RR, MC-YR, and MC-LR and were 0.1 and 0.5 microg g(-1), respectively. MCs recoveries at three levels in spiked samples (0.5-3.0 microg g(-1)) were > 92%, with relative standards deviations (RSDs) < 16% for liver samples and > 68% with RSDs < 18% for intestine samples. The proposed method was applied to determine MC-LR in exposed fish to evaluate the bioaccumulation risk. The results showed the transference of MC-LR from cyanobacterial cells to fish tissues.

Animals↗

FAS/FAS ligand ratio: a marker of oxaliplatin-based intrinsic and acquired resistance in advanced colorectal cancer.

PURPOSE: Oxaliplatin-5-fluorouracil combinations have increased responses in first-line therapy up to 40% in advanced colorectal cancer. Unfortunately, those patients who will respond are unknown and initially sensitive patients become rapidly resistant to current therapies. FAS (CD95) and FAS ligand (FASL; CD95L) have been implicated in chemosensitivity through leading to apoptosis in response to DNA-damaging drugs. Whereas the proapoptotic role of FAS and FASL is well characterized, the function of their soluble forms as predictors of chemosensitivity remains unknown. PATIENTS AND METHODS: Blood samples were obtained from 68 patients with advanced colorectal cancer who received oxaliplatin-5-fluorouracil combinations in first-line therapy. Computed tomographic scans were done every 3 months and responses were evaluated by Response Evaluation Criteria in Solid Tumors criteria. ELISA soluble FAS and soluble FASL analysis were done before treatment and every 3 months until disease progression. Ratios between soluble FAS and soluble FASL were established and its values and variations through time were related to treatment responses. RESULTS: We found a significant increase in soluble FAS levels and a significant decrease in FASL at 3 months compared with baseline (13.2 versus 10.02 ng/mL; P=0.0001; 0.07 versus 0.14 ng/mL; P=0.007, respectively). A significant increase in the soluble FASL levels up to 9 months (fourth to fifth extractions; 0.26 ng/mL) of therapy compared with first to third extractions (0.11 ng/mL; P=0.003) was also found. A random effect regression statistical model determined that >1.2-fold increase in soluble FAS/soluble FASL ratio was a marker of chemosensitivity (P = 0.001). CONCLUSIONS: These data strongly indicate that an increment of soluble FAS/soluble FASL ratio after treatment could be an excellent marker of chemosensitivity in colorectal cancer. On the other hand, a decreased ratio after treatment can be a predictor of chemoresistance despite an initial response.

Adult↗

Probing the determinants of substrate specificity of a feruloyl esterase, AnFaeA, from Aspergillus niger.

Feruloyl esterases hydrolyse phenolic groups involved in the cross-linking of arabinoxylan to other polymeric structures. This is important for opening the cell wall structure making material more accessible to glycoside hydrolases. Here we describe the crystal structure of inactive S133A mutant of type-A feruloyl esterase from Aspergillus niger (AnFaeA) in complex with a feruloylated trisaccharide substrate. Only the ferulic acid moiety of the substrate is visible in the electron density map, showing interactions through its OH and OCH(3) groups with the hydroxyl groups of Tyr80. The importance of aromatic and polar residues in the activity of AnFaeA was also evaluated using site-directed mutagenesis. Four mutant proteins were heterologously expressed in Pichia pastoris, and their kinetic properties determined against methyl esters of ferulic, sinapic, caffeic and p-coumaric acid. The k(cat) of Y80S, Y80V, W260S and W260V was drastically reduced compared to that of the wild-type enzyme. However, the replacement of Tyr80 and Trp260 with smaller residues broadened the substrate specificity of the enzyme, allowing the hydrolysis of methyl caffeate. The role of Tyr80 and Trp260 in AnFaeA are discussed in light of the three-dimensional structure.

Aspergillus niger↗

Induction of NAD(P)H quinone oxidoreductase by vegetables widely consumed in Catalonia, Spain.

Monofunctional inducers (MIs) enhance phase 2 enzymes such as nicotinamide-adenine-dinucleotide-phosphate [NAD(P)H] quinone oxidoreductase (NQO1) without modifying oxidation enzymes. The induction of these protective enzymes appears to be mediated by genetic regulatory elements in their promoter regions known as the antioxidant response element (ARE). The aim of this study was to identify, through an in vitro study, which of the 30 fruits and vegetables commonly consumed in Catalonia, Spain, contain MIs of NQO1. We assayed the capacity of extracts of these fruits and vegetables to induce NQO1 [by more than 1.5-fold: ratio of induction (cells treated/control) >1.5, 8-mg/ml dose] in two murine hepatoma cell lines: Hepa 1c1c7 and BPrC1, a modified cell line that possesses a nonfunctional aryl hydrocarbon receptor nuclear translocator system and is thus nonresponsive to bifunctional inducers. We also used a third cell line, papiloma (PE) murine keratinocytes, a stably transfected cell line with an ARE-luc+ plasmid (AREPE cell line) for verifying induction through the ARE with a simple luminescence screening assay. Broccoli (Hepa 1c1c7, ratio=5.5; BPrC1, ratio=2.3), calcot (Allium cepa L.) (Hepa 1c1c7, ratio=4.7; BPrC1, ratio=.5), green onion (Hepa 1c1c7, ratio=4.6; BPrC1, ratio=2), green cabbage (Hepa 1c1c7, ratio=3.6; BPrC1, ratio=2.7), purple cabbage (Hepa 1c1c7, ratio=3.4; BPrC1, ratio=2), and black cabbage (Hepa 1c1c7, ratio=3; BPrC1, ratio=3) were active NQO1 inducers in both murine hepatoma cell lines. Extracts from broccoli (ratio=3.5), calcot (ratio=4.8), cauliflower (ratio=4.2), cabbage (ratio=2.2), green onion (ratio=3.2), green cabbage (ratio=3.6), black cabbage (ratio=4.5), and purple cabbage (ratio=3.7) were confirmed to contain MIs in the AREPE cell line. These results are very similar to those described for vegetables consumed in the United States, with the exception of calcot, which is common in Catalonia but is not grown or consumed widely in the United States.

Animals↗

The crystal structure of feruloyl esterase A from Aspergillus niger suggests evolutive functional convergence in feruloyl esterase family.

As a component of the array of enzymes produced by micro-organisms to deconstruct plant cell walls, feruloyl esterases hydrolyze phenolic groups involved in the cross-linking of arabinoxylan to other polymeric structures. This is important for opening the cell wall structure, making material more accessible to glycosyl hydrolases. Here, we describe the first crystal structure of the non-modular type-A feruloyl esterase from Aspergillus niger (AnFaeA) solved at 2.5A resolution. AnFaeA displays an alpha/beta hydrolase fold similar to that found in fungal lipases and different from that reported for other feruloyl esterases. Crystallographic and site-directed mutagenesis studies allow us to identify the catalytic triad (Ser133-His247-Asp194) that forms the catalytic machinery of this enzyme. The active-site cavity is confined by a lid (residues 68-80), on the analogy of lipases, and by a loop (residues 226-244) that confers plasticity to the substrate-binding site. The lid presents a high ratio of polar residues, which in addition to a unique N-glycosylation site stabilises the lid in an open conformation, conferring the esterase character to this enzyme. A putative model for bound 5,5'-diferulic acid-linked arabinoxylan has been built, pointing to the more relevant residues involved in substrate recognition. Comparison with structurally related lipases reveals that subtle amino acid and conformational changes within a highly conserved protein fold may produce protein variants endowed with new enzymatic properties, while comparison with functionally related proteins points to a functional convergence after evolutionary divergence within the feruloyl esterases family.

Aspergillus niger↗

[Usefulness of PSA and its fractions in the diagnosis of prostate cancer].

BACKGROUND AND OBJECTIVE: The description of different forms of PSA has opened a new strategy in the diagnosis of prostate cancer. The measurement of the ratio between free PSA and PSA in the group of patients with a PSA level between 4 and 10 ng/ml decreases the number of negative biopsies. The aim of our study was to compare the diagnostic efficacy of PSA and PSA fractions (free PSA [fPSA] and complexed PSA [cPSA]) in the differential diagnosis between Pca and benign prostate hyperplasia (BPH). PATIENTS AND METHOD: We measured the serum levels of PSA, free PSA and cPSA in 56 patients with Pca and 94 patients with BPH. RESULTS: ROC curves were used for the comparison of tests. The biggest area under the curve (AUC) was observed for the ratios fPSA/cPSA and fPSA/PSA (0.718 and 0.712, respectively). When we compared the AUC between PSA and cPSA, then AUC for cPSA was higher than AUC for PSA (0.602 and 0.567, respectively). We observed similar results in the group of patients with PSA levels between 4 and 10 ng/ml. CONCLUSIONS: The diagnostic accuracy of cPSA is higher than that of PSA. Moreover, in the differential diagnosis between prostate cancer and BPH, the use of PSA ratios (fPSA/cPSA or fPSA/PSA) increases the diagnostic accuracy obtained with the measurement of PSA or cPSA.

Biomarkers↗

ProGRP: a new biomarker for small cell lung cancer.

Progastrin-releasing peptide (ProGRP) is a recently identified biomarker of small cell lung cancer (SCLC), a disorder of neuroendocrine tissue differentiation. The upper normal limit of ProGRP in the circulation is 50 pg/ml. Impaired glomerular filtration tends to increase circulating levels and confound the tumor marker significance of modestly elevated values. Excluding patients with renal failure, circulating levels did not exceed 80 pg/ml in benign disease (3% of cases in excess of the upper normal limit) or 120 pg/ml in malignancy other than lung cancer and neuroendocrine tumors (5% of cases in excess of the upper limit). ProGRP serum levels are clearly related to the lung cancer histological type with significantly higher levels observed in SCLC than in nonsmall cell lung cancer (NSCLC). Circulating ProGRP in excess of 120 mg/ml was found in only 4% of cases of NSCLC with another 22% presenting with modestly elevated levels in excess of the upper normal limit. By contrast, abnormal ProGRP results are found in 60-70% and in 75-90% of SCLC patients with local and extensive disease, respectively. ProGRP is a more sensitive biomarker than is neuron-specific enolase (NSE) for SCLC, but thus far has not been found in multivariate analysis to have independent prognostic significance. Preliminary studies suggest ProGRP will have utility in conjunction with NSE in monitoring the therapy of established SCLC.

Biomarkers, Tumor↗

Comparison of several combinations of free, complexed, and total PSA in the diagnosis of prostate cancer in patients with urologic symptoms.

OBJECTIVES: To compare the diagnostic efficacy of prostate-specific antigen (PSA) and the PSA fractions (free PSA [fPSA] and complexed PSA [cPSA]) in the differential diagnosis between prostate cancer and benign prostatic hyperplasia. METHODS: We measured the serum levels of total PSA (tPSA; Hybritech and Bayer), fPSA (Hybritech), and cPSA (Bayer) in 72 patients with prostate cancer and 128 patients with benign prostatic hyperplasia. RESULTS: Receiver operating characteristic curves were used for comparison of these tests. The greatest area under the curve was observed for the fPSA/cPSA ratio and the fPSA/tPSA ratio (0.757 and 0.754, respectively). The substitution of the fPSA/tPSA ratio with the fPSA/cPSA ratio in the diagnostic scheme of prostate cancer improved the diagnostic accuracy, with similar sensitivity and an increment in specificity (41% versus 45%). CONCLUSIONS: The fPSA/cPSA ratio ensures a reduction in negative biopsies in the PSA gray zone. We suggest substituting the fPSA/tPSA ratio with the fPSA/cPSA ratio for patients with a PSA level between 4 and 10 ng/mL.

Aged↗

Parasitic helminths in the digestive tract of six species of owls (Strigiformes) in Spain.

A survey was carried out in Catalonia, Spain to determine the prevalence of parasitic helminths infesting the digestive tract of nocturnal raptors (Strigiformes). One hundred birds belonging to six owl species were examined. The number of birds examined ranged between three and 30. The overall level of infestation was 65% and the most frequent helminths present were nematodes, followed by trematodes, cestodes and acanthocephalans. Strigiformes showing the highest levels of infestation were little owls (Athene noctua), 86.7% of which were infested. Helminths with the highest prevalence were Synhimantus spp., intestinal capillarids, Brachylaima spp., cestodes and Centrorhynchus aluconis.

Animals↗

Bayesian resolution enhancement of compressed video.

Super-resolution algorithms recover high-frequency information from a sequence of low-resolution observations. In this paper, we consider the impact of video compression on the super-resolution task. Hybrid motion-compensation and transform coding schemes are the focus, as these methods provide observations of the underlying displacement values as well as a variable noise process. We utilize the Bayesian framework to incorporate this information and fuse the super-resolution and post-processing problems. A tractable solution is defined, and relationships between algorithm parameters and information in the compressed bitstream are established. The association between resolution recovery and compression ratio is also explored. Simulations illustrate the performance of the procedure with both synthetic and nonsynthetic sequences.

Algorithms↗

Pilot trial of trastuzumab starting with or after the doxorubicin component of a doxorubicin plus paclitaxel regimen for women with HER2-positive advanced breast cancer.

PURPOSE: Combining trastuzumab with doxorubicin and paclitaxel (AT) is attractive because of the activity of AT and survival improvements observed when trastuzumab is added to either agent in HER2-positive metastatic breast cancer. This pilot study evaluates the efficacy and cardiac tolerability of AT followed by paclitaxel with trastuzumab started with AT or paclitaxel alone and investigates pharmacokinetic interactions. EXPERIMENTAL DESIGN: Two cohorts of 16 patients were enrolled. Cohort 1 received three cycles of AT (60/150 mg/m2) plus trastuzumab (4 mg/kg initial dose followed by 2 mg/kg), initiated concomitantly with doxorubicin, followed by nine cycles of paclitaxel (80 mg/m2) plus trastuzumab and then trastuzumab alone. Cohort 2 was treated with the same regimen, but trastuzumab was initiated with paclitaxel after AT. Cardiac function, pharmacokinetic interactions, and efficacy were evaluated. RESULTS: Median baseline left ventricular ejection fraction (LVEF) was 62% (range, 57-74%) and 66% (range, 57-77%) in cohorts 1 and 2, respectively. Most patients had an absolute decrease in LVEF. Congestive heart failure was not observed. LVEF in three patients decreased to <50% but recovered despite continued treatment. Response rates were 87.5% in both cohorts (cohort 1:2 complete response, 12 partial response; cohort 2:3 complete response, 11 partial response). No unexpected side effects were observed. Pharmacokinetics of paclitaxel and its metabolites and of doxorubicin were similar without and with trastuzumab. CONCLUSIONS: Trastuzumab administered with AT followed by weekly paclitaxel alone is highly active whether trastuzumab is initiated with AT or paclitaxel. Congestive heart failure was not observed, and LVEF decreases were reversible. Further studies of this regimen are warranted.

Adult↗

Clinical evaluation of the Elecsys CA 15-3 test in breast cancer patients.

The aim of the present study was to evaluate the clinical performance of the CA 15-3 assay on Elecsys systems in an international multicenter study (11 centers). A total of 1326 single samples (272 apparently healthy individuals, 34 pregnant women, 308 benign diseases, 273 cancers other than breast, 439 breast cancer) and 538 serial samples of 98 breast cancer patients during follow-up were analyzed. 95% of values in healthy individuals were below 25 kU/L, and 88% in benign breast diseases, respectively. In malignant breast disease at primary diagnosis the value distribution of Elecsys CA 15-3, sensitivity at 95% specificity, as well as the areas under the curve in ROC analysis were clearly correlated to tumor stages: UICC I to IV 88 to 25% of values < 25 kU/L, sensitivity 7 to 78%, areas under the curve 0.53 to 0.94. During follow-up, sensitivity/specificity for detection of recurrences were 90%/71%. In metastatic disease clinical progression/response to therapy were indicated in 91%/78% of patients at a specificity of 92%/78%. The findings indicate that the Elecsys CA 15-3 assay is very suitable in routine work for detection of recurrences as well as for therapy control in metastatic breast cancer.

Adolescent↗

S-100 protein serum levels in patients with benign and malignant diseases: false-positive results related to liver and renal function.

S-100 serum concentrations were analyzed in 39 healthy people, 130 patients with benign diseases and 304 patients with malignancies, including 49 patients with locoregional diseases and 255 with advanced diseases. S-100 was determined by a commercial immunoluminometric assay, and 0.20 ng/ml was considered to be the upper limit of normality. In none of the healthy people was S-100 higher than 0.2 ng/ml. Slightly high S-100 concentrations were found in 33 out of 130 patients (25%) with benign diseases (mean 0.21 +/- 0.45 ng/ml). Significantly higher S-100 serum levels were found in patients with liver cirrhosis (63%, 10/16) (p = 0.024) or renal failure (45%, 8/18) (p = 0.03) than in patients with other benign diseases or in healthy people. Abnormal S-100 serum levels were found in 68 of the patients (22.5%) with malignancy (mean 1.01 +/- 5.9 ng/ml). The highest S-100 concentrations were found in patients with malignant melanomas (p = 0.001). Excluding melanoma patients, the S-100 serum levels in patients with malignancies were not related to tumor origin or stage but were clearly related to the site of metastasis, with patients with liver metastases showing higher values than patients with metastases without liver involvement (p = 0.02). No statistical differences were found among patients with liver cirrhosis, primary liver cancer or liver metastases. In conclusion, S-100 is a useful marker for melanoma, but abnormal levels of this tumor marker may be found in benign and malignant diseases associated with liver or renal injury.

Adult↗

Prospective evaluation of squamous cell carcinoma and carcinoembryonic antigen as prognostic factors in patients with cervical cancer.

Carcinoembryonic antigen (CEA) and squamous cell carcinoma(SCC) serum levels were prospectively determined in 159 untreated patients diagnosed with carcinoma of the uterine cervix from 1991 to 2001. The histological analysis showed epidermoid cancer in 117 patients, adenocarcinoma in 26 patients, adenosquamous carcinoma in 12 patients and other histological types in the remaining 4 patients. Tumor marker sensitivity was related to the histological type with abnormal SCC (>2 ng/ml) in 51.3% of squamous tumors in contrast to the 7.1% found in other histologies. By contrast, CEA sensitivity was not related to histology with abnormal values (>5 ng/ml) in 25% of squamous tumors, 19% of adenocarcinomas, 33% of adenosquamous carcinomas and 25% of other histologies. CEA and SCC serum levels were clearly related to tumor stage, parametrial invasion, tumor size and nodal involvement. Elevated pretreatment CEA indicates parametrial invasion with a probability of 82%. Likewise, pretreatment CEA and SCC serum levels were of prognostic value, with a shorter disease-free survival and overall survival in patients with abnormal levels. All patients with adenocarcinomas and abnormal CEA had relapse during follow-up. Multivariate analysis indicated that parametrial invasion, age, tumor size and SCC were independent prognostic factors. In conclusion, CEA and SCC are useful tumor markers in carcinomas of the uterine cervix, with a clear relationship with well-known prognostic factors (parametrial invasion, nodal involvement), and are of prognostic value.

Adenocarcinoma↗

Relationship of p53 with other oncogenes, cytokines and systemic lupus erythematosus activity.

INTRODUCTION: Defects in the regulation of apoptosis of autoreactive lymphocytes are involved in the pathogenesis of systemic lupus erythematosus (SLE). The apoptotic process relies on adequate functioning of numerous molecules, including oncogenes and diverse cytokines. p53 has been implicated in the control of the cell cycle through the stimulation of apoptosis of these autoreactive cells. OBJECTIVE: To study the role of the p53 pathway on the regulation of apoptosis in SLE patients and analyze the relationship of the p53 oncoprotein with disease activity and other oncogenes (bcl-2, Fas) and cytokines (interleukin-10, IL-10, and tumor necrosis factor-alpha, TNF-alpha), implicated in the apoptotic process and the pathogenesis of SLE. PATIENTS AND METHODS: p53 and bcl-2 antigen expression were determined in lyzed lymphocytes from 74 patients with SLE and 30 healthy controls. Serum levels of soluble-Fas (sFas) and cytokines IL-10 and TNF-alpha were studied by enzyme-linked immunonosorbent assay. RESULTS: SLE patients had higher levels of p53 protein (0.16 +/- 0.33 ng/dl) than controls (0.014 +/- 0.02 ng/dl; p = 0.006). Patients with active SLE had higher levels of p53 (0.31 +/- 0.48 ng/dl) than those with inactive disease (0.08 +/- 0.17 ng/dl; p = 0.003) who in turn had higher levels than controls (0.01 +/- 0.02 ng/dl; p = 0.035). A significant correlation was found between p53 levels and the SLE disease activity index (R = +0.24/ p = 0.04), anti-DNA antibodies (R = +0.23/p = 0.048) and IL-10 levels (R = +0.4/p = 0.004). No correlation was found between p53 levels and bcl-2, sFas or TNF-alpha levels. CONCLUSIONS: The p53 oncoprotein may play a role in the pathogenesis and activity of SLE. IL-10 may influence SLE activity by inhibiting the p53 and bcl-2/Fas apoptosis pathway of autoreactive cells.

Adolescent↗