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1.
Anal Bioanal Chem ; 404(5): 1317-26, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22543714

ABSTRACT

This paper discusses gene expression changes in the skin of mice treated by monoenergetic 14 MeV neutron irradiation and the possibility of monitoring the resultant lipid depletion (cross-validated by functional genomic analysis) as a marker of radiation exposure by high-resolution FT-IR (Fourier transform infrared) imaging spectroscopy. The irradiation was performed at the ENEA Frascati Neutron Generator (FNG), which is specifically dedicated to biological samples. FNG is a linear electrostatic accelerator that produces up to 1.0 × 10(11) 14-MeV neutrons per second via the D-T nuclear reaction. The functional genomic approach was applied to four animals for each experimental condition (unirradiated, 0.2 Gy irradiation, or 1 Gy irradiation) 6 hours or 24 hours after exposure. Coregulation of a subclass of keratin and keratin-associated protein genes that are physically clustered in the mouse genome and functionally related to skin and hair follicle proliferation and differentiation was observed. Most of these genes are transiently upregulated at 6 h after the delivery of the lower dose delivered, and drastically downregulated at 24 h after the delivery of the dose of 1 Gy. In contrast, the gene coding for the leptin protein was consistently upregulated upon irradiation with both doses. Leptin is a key protein that regulates lipid accumulation in tissues, and its absence provokes obesity. The tissue analysis was performed by monitoring the accumulation and the distribution of skin lipids using FT-IR imaging spectroscopy. The overall picture indicates the differential modulation of key genes during epidermis homeostasis that leads to the activation of a self-renewal process at low doses of irradiation.


Subject(s)
Leptin/metabolism , Neutrons , Skin/metabolism , Skin/radiation effects , Spectroscopy, Fourier Transform Infrared , Animals , Leptin/analysis , Lipid Metabolism , Lipids/analysis , Male , Mice , Mice, Inbred C57BL , Radiation Dosage
2.
Curr Cancer Drug Targets ; 9(2): 118-30, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19275753

ABSTRACT

The natural polyamines (PA), putrescine (PUT), spermidine (SPD) and spermine (SPM) are ubiquitous constituents of eukaryotic cells. The increase of PA in malignant and proliferating cells attracted the interest of scientists during last decades, addressing PA depletion as a new strategy to inhibit cell growth. Selective enzyme inhibitors were developed for decreasing PA metabolism and to act as chemotherapeutic anticancer agents. Indeed, the complexity of the PA homoeostasis overcomes the PA perturbation by a single enzyme to take effect therapeutically. Recently, an increasing interest has been posed on spermine-oxidase (SMO), the only catabolic enzyme able to specifically oxidise SPM. Interestingly, the absence of SPM is compatible with life, but its accumulation and degradation is lethal. Augmented SMO activity provokes an oxidative stress rendering cells prone to die, and appears to be important in the cell differentiation pathway. Extra-cellular SPM is cytotoxic, but its analogues are capable of inhibiting cell growth at low concentrations, most likely by intracellular SPM depletion. These pivotal roles seem to evoke the biological processes of stress response, wherein balance is mandatory to live or to die. Thus, altering SPM metabolism could allow a multi-tasking therapeutic strategy, addressed not only to inhibit PA metabolism. Several tetramines are presently in early phases (I and II) of clinical trials, and it will be a matter of a few more years to understand whether SPM-related therapeutic approaches would be of benefit for composite treatment protocols of cancer.


Subject(s)
Antineoplastic Agents/pharmacology , Spermine/antagonists & inhibitors , Spermine/metabolism , Animals , Biogenic Polyamines/metabolism , Enzyme Inhibitors/pharmacology , Homeostasis/drug effects , Homeostasis/physiology , Humans , Inflammation/physiopathology , Models, Molecular , Neoplasms/drug therapy , Oxidoreductases Acting on CH-NH Group Donors/antagonists & inhibitors , Oxidoreductases Acting on CH-NH Group Donors/metabolism , Protein Conformation , Spermine/analogs & derivatives , Polyamine Oxidase
3.
Biochim Biophys Acta ; 1773(6): 774-83, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17363080

ABSTRACT

In the aging process and in most degenerative diseases, the oxidant by-products of cellular metabolism lead to oxidative stress. Oxidative stress plays an important role in switching from cell proliferation to its opposite outcome, cell death. The metabolic pathways in charge of the interconversion and degradation of the polyamines are responsible for oxidant by-products. In the past few years, spermine metabolism has been found closely related to DNA oxidation and apoptosis. Moreover, that the ectopical expression of murine spermine oxidase induced DNA damage in the neuroblastoma cell line, and this was uncoupled with any increase in cell mortality, thus suggests an activation of DNA repair. In this work, we provide new evidence showing that only spermine oxidase overactivity can deliver sub-lethal chronic DNA damage and repair without affecting transcriptional and enzymatic levels of the PA key regulatory enzymes ODC and SSAT. Chronic sub-lethal DNA damage is below the cell cycle arrest induction threshold, but is able to activate apurinic/apyrimidinic endonuclease protein (APE1) and gamma H2AX. Of therapeutic interest, the chronic sub-lethal DNA damage and activation of the repair processes are in turn responsible for inducing hypersensitivity after exposure to radiation with no induction of adaptive response to damage.


Subject(s)
Cell Proliferation/radiation effects , DNA Damage/radiation effects , DNA Repair/radiation effects , Oxidative Stress/radiation effects , Oxidoreductases Acting on CH-NH Group Donors/biosynthesis , X-Rays , Aging/metabolism , Aging/radiation effects , Animals , Cell Death/radiation effects , Cell Line, Tumor , DNA Damage/genetics , DNA Repair/genetics , DNA-(Apurinic or Apyrimidinic Site) Lyase/metabolism , Histones/metabolism , Mice , Oxidoreductases Acting on CH-NH Group Donors/genetics , Polyamine Oxidase
4.
Prensa méd. argent ; 92(10): 657-665, dic. 2005. tab
Article in Spanish | LILACS | ID: lil-427093

ABSTRACT

Hemos planteado el presente trabajo, teniendo en cuenta los siguientes aspectos: describir las características epidemiológicas y demográficas en la población estudiada y además caracterizar especialmente los factores higiénicos, sanitarios y culturales que por su heterogeneidad, tuvieron influencia en la capacidad de infección por el Helicobacter Pylori


Subject(s)
Adolescent , Adult , Humans , Aged , Demography , Educational Status , Epidemiologic Studies , Health Status Indicators , Helicobacter pylori , Economic Indexes/statistics & numerical data , Quality of Life , Social Class
5.
Prensa méd. argent ; 92(10): 657-665, dic. 2005. tab
Article in Spanish | BINACIS | ID: bin-411

ABSTRACT

Hemos planteado el presente trabajo, teniendo en cuenta los siguientes aspectos: describir las características epidemiológicas y demográficas en la población estudiada y además caracterizar especialmente los factores higiénicos, sanitarios y culturales que por su heterogeneidad, tuvieron influencia en la capacidad de infección por el Helicobacter Pylori


Subject(s)
Adolescent , Adult , Humans , Aged , Helicobacter pylori/classification , Epidemiologic Studies , Quality of Life , Social Class , Economic Indexes/statistics & numerical data , Health Status Indicators , Quality of Life , Educational Status , Demography
6.
Biochim Biophys Acta ; 1755(1): 15-24, 2005 May 25.
Article in English | MEDLINE | ID: mdl-15907589

ABSTRACT

In mammals, the polyamines affect cell growth, differentiation, and apoptosis; their levels are increased in malignant and proliferating cells, thus justifying an interest in a chemotherapeutic approach to cancer. The flavoprotein SMO is the most recently characterized catabolic enzyme, preferentially oxidizing SPM to SPD, 3-aminopropanal and H(2)O(2). In this report, we describe a novel functional characterization of the recently cloned splice variant isoforms from mouse brain, encoding, among others, the nuclear co-localized spermine oxidase mSMOmu. The over-expression of the active isoforms mSMOalpha and mSMOmu, and the inactive mSMOdelta and mSMOgamma in mouse neuroblastoma cells, demonstrated the first evidence of the direct oxidative DNA damage by the SMO activities, either alone or, in a higher extent, when associated with radiation exposure, thus working as radio sensitizer. These effects were reverted by treatment with 50 muM and 100 muM doses of the inhibitor of SMO activity MDL 72,527. The over-expression of all SMO isoforms failed to influence the expression of the regulating enzymes of polyamines metabolism ODC and SSAT. Dealing with the unbalanced tissue specific SMO activities, these results could indicate a new direction to tailor chemotherapy-associated radiotherapy, improving dose-rate protocol and allowing the modulation of deleterious side effects on healthy tissues.


Subject(s)
Apoptosis , DNA Damage , Guanine/analogs & derivatives , Neuroblastoma/metabolism , Oxidoreductases Acting on CH-NH Group Donors/metabolism , Radiation Tolerance , Animals , Guanine/metabolism , Isoenzymes/metabolism , Mice , Oxidation-Reduction , Polyamines/metabolism , Tumor Cells, Cultured , Polyamine Oxidase
7.
Acta Gastroenterol Latinoam ; 33(4): 199-203, 2003.
Article in Spanish | MEDLINE | ID: mdl-14708472

ABSTRACT

OBJECTIVE: To compare the acid-supressing capacity of omeprazole (OZ) 20 mg tablets vs pantoprazole (PZ) 20 and 40 mg tablets, in healthy volunteers, with 24-h intragastric pH-metry. MATERIAL AND METHODS: Open, randomized, cross-over trial in 10 healthy volunteers; on days 0.8 and 22, 24-h intragastric pH-metry. Day 0, basal, thereafter 7 days with OZ or PZ 20 mg/day; day 8, pH-metry, then "wash out" for 7 days and thereafter 7 more days' therapy with PZ or OZ. On day 22 a 24-h intragastric pH control was performed again. In the last treatment stage, all of them were administered pantoprazole 40 mg/day for 8 days again with a 24-h pH recording at the end. RESULTS: 24-h pH-metry expressed as the time (hours) in which the pH was < or = 4 and the values as mean +/- standard deviation. BASAL 22.12 +/- 1.54, POST-OZ 9.78 +/- 6.72, POST-PZ 20 15.65 +/- 5.65, POST-PZ 40 8.57 +/- 5.93. Statistical evaluation with two way repeated measures ANOVA p < 0.0001. Newman Keuls post-hoc test: (1) vs (2) p < 0.003; (1) vs (3) p < 0.03; (2) vs (4) 0.65. CONCLUSIONS: According to the results it might be stated that both proton pump inhibitors have acid-supressing capacity and omeprazole in equal dosis is more effective than pantoprazole as acid-supressor, with statistically significative differences. There was no difference between 20 mg omeprazole and 40 mg pantoprazole.


Subject(s)
Anti-Ulcer Agents/pharmacology , Benzimidazoles/pharmacology , Gastric Acid/metabolism , Omeprazole/pharmacology , Proton Pump Inhibitors , Sulfoxides/pharmacology , 2-Pyridinylmethylsulfinylbenzimidazoles , Adult , Analysis of Variance , Cross-Over Studies , Female , Humans , Hydrogen-Ion Concentration , Male , Middle Aged , Pantoprazole , Time Factors
8.
Acta gastroenterol. latinoam ; 33(4): 199-203, 2003. tab
Article in Spanish | LILACS | ID: lil-359984

ABSTRACT

OBJECTIVE: To compare the acid-supressing capacity of omeprazole (OZ) 20 mg tablets vs pantoprazole (PZ) 20 and 40 mg tablets, in healthy volunteers, with 24-h intragastric pH-metry. MATERIAL AND METHODS: Open, randomized, cross-over trial in 10 healthy volunteers; on days 0.8 and 22, 24-h intragastric pH-metry. Day 0, basal, thereafter 7 days with OZ or PZ 20 mg/day; day 8, pH-metry, then "wash out" for 7 days and thereafter 7 more days' therapy with PZ or OZ. On day 22 a 24-h intragastric pH control was performed again. In the last treatment stage, all of them were administered pantoprazole 40 mg/day for 8 days again with a 24-h pH recording at the end. RESULTS: 24-h pH-metry expressed as the time (hours) in which the pH was < or = 4 and the values as mean +/- standard deviation. BASAL 22.12 +/- 1.54, POST-OZ 9.78 +/- 6.72, POST-PZ 20 15.65 +/- 5.65, POST-PZ 40 8.57 +/- 5.93. Statistical evaluation with two way repeated measures ANOVA p < 0.0001. Newman Keuls post-hoc test: (1) vs (2) p < 0.003; (1) vs (3) p < 0.03; (2) vs (4) 0.65. CONCLUSIONS: According to the results it might be stated that both proton pump inhibitors have acid-supressing capacity and omeprazole in equal dosis is more effective than pantoprazole as acid-supressor, with statistically significative differences. There was no difference between 20 mg omeprazole and 40 mg pantoprazole.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Benzimidazoles/pharmacology , Enzyme Inhibitors/pharmacology , Gastric Acid , Omeprazole/pharmacology , Proton Pumps/antagonists & inhibitors , Sulfoxides/pharmacology , Analysis of Variance , Benzimidazoles/administration & dosage , Cross-Over Studies , Enzyme Inhibitors/administration & dosage , Gastric Acidity Determination , Hydrogen-Ion Concentration , Manometry , Omeprazole/administration & dosage , Random Allocation , Sulfoxides/administration & dosage , Time Factors
9.
Acta gastroenterol. latinoam ; 33(4): 199-203, 2003. tab
Article in Spanish | BINACIS | ID: bin-4798

ABSTRACT

OBJECTIVE: To compare the acid-supressing capacity of omeprazole (OZ) 20 mg tablets vs pantoprazole (PZ) 20 and 40 mg tablets, in healthy volunteers, with 24-h intragastric pH-metry. MATERIAL AND METHODS: Open, randomized, cross-over trial in 10 healthy volunteers; on days 0.8 and 22, 24-h intragastric pH-metry. Day 0, basal, thereafter 7 days with OZ or PZ 20 mg/day; day 8, pH-metry, then "wash out" for 7 days and thereafter 7 more days therapy with PZ or OZ. On day 22 a 24-h intragastric pH control was performed again. In the last treatment stage, all of them were administered pantoprazole 40 mg/day for 8 days again with a 24-h pH recording at the end. RESULTS: 24-h pH-metry expressed as the time (hours) in which the pH was < or = 4 and the values as mean +/- standard deviation. BASAL 22.12 +/- 1.54, POST-OZ 9.78 +/- 6.72, POST-PZ 20 15.65 +/- 5.65, POST-PZ 40 8.57 +/- 5.93. Statistical evaluation with two way repeated measures ANOVA p < 0.0001. Newman Keuls post-hoc test: (1) vs (2) p < 0.003; (1) vs (3) p < 0.03; (2) vs (4) 0.65. CONCLUSIONS: According to the results it might be stated that both proton pump inhibitors have acid-supressing capacity and omeprazole in equal dosis is more effective than pantoprazole as acid-supressor, with statistically significative differences. There was no difference between 20 mg omeprazole and 40 mg pantoprazole.(AU)


Subject(s)
Comparative Study , Humans , Male , Female , Adult , Middle Aged , Benzimidazoles/pharmacology , Omeprazole/pharmacology , Gastric Acid/metabolism , Enzyme Inhibitors/pharmacology , Proton Pumps/antagonists & inhibitors , Sulfoxides/pharmacology , Benzimidazoles/administration & dosage , Omeprazole/administration & dosage , Cross-Over Studies , Hydrogen-Ion Concentration , Enzyme Inhibitors/administration & dosage , Time Factors , Gastric Acidity Determination , Random Allocation , Analysis of Variance , Manometry , Sulfoxides/administration & dosage
10.
Acta gastroenterol. latinoam ; 33(4): 199-203, 2003.
Article in Spanish | BINACIS | ID: bin-38809

ABSTRACT

OBJECTIVE: To compare the acid-supressing capacity of omeprazole (OZ) 20 mg tablets vs pantoprazole (PZ) 20 and 40 mg tablets, in healthy volunteers, with 24-h intragastric pH-metry. MATERIAL AND METHODS: Open, randomized, cross-over trial in 10 healthy volunteers; on days 0.8 and 22, 24-h intragastric pH-metry. Day 0, basal, thereafter 7 days with OZ or PZ 20 mg/day; day 8, pH-metry, then [quot ]wash out[quot ] for 7 days and thereafter 7 more days therapy with PZ or OZ. On day 22 a 24-h intragastric pH control was performed again. In the last treatment stage, all of them were administered pantoprazole 40 mg/day for 8 days again with a 24-h pH recording at the end. RESULTS: 24-h pH-metry expressed as the time (hours) in which the pH was < or = 4 and the values as mean +/- standard deviation. BASAL 22.12 +/- 1.54, POST-OZ 9.78 +/- 6.72, POST-PZ 20 15.65 +/- 5.65, POST-PZ 40 8.57 +/- 5.93. Statistical evaluation with two way repeated measures ANOVA p < 0.0001. Newman Keuls post-hoc test: (1) vs (2) p < 0.003; (1) vs (3) p < 0.03; (2) vs (4) 0.65. CONCLUSIONS: According to the results it might be stated that both proton pump inhibitors have acid-supressing capacity and omeprazole in equal dosis is more effective than pantoprazole as acid-supressor, with statistically significative differences. There was no difference between 20 mg omeprazole and 40 mg pantoprazole.

11.
Acta gastroenterol. latinoam ; 32(2): 83-85, nov. 2002.
Article in Spanish | LILACS | ID: lil-327728

ABSTRACT

Nowadays technics for Helicobacter pylori detection in stools like culture, and PCR, are expensive and difficult to perform. The aim of this study was to evaluate ELISA test efficacy for detection of H. Pylori antigens in stools comparing this results with standarized technics like histology (Giemsa), ureasa test and UBT C 14. 26 patients were evaluated in this study, ages between 15-75 with upper gastrointestinal symptoms; all of them required gastroduodenal endoscopy, status H. Pylori was determined with methods upon mentioned. 24 hours after endoscopy H. Pylori antigens in stools with the technique Premier Platinum Htsa, Elisa were determined. The detection of H. Pylori antigens in stools accurately identified active H. Pylori infection. The performance characteristics of this non-invasive method was similar in sensibility and specificity to conventional tests


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Antigens, Bacterial , Feces , Helicobacter Infections , Helicobacter pylori , Immunoenzyme Techniques , Enzyme-Linked Immunosorbent Assay , Sensitivity and Specificity
12.
Acta gastroenterol. latinoam ; 32(2): 83-85, nov. 2002.
Article in Spanish | BINACIS | ID: bin-7030

ABSTRACT

Nowadays technics for Helicobacter pylori detection in stools like culture, and PCR, are expensive and difficult to perform. The aim of this study was to evaluate ELISA test efficacy for detection of H. Pylori antigens in stools comparing this results with standarized technics like histology (Giemsa), ureasa test and UBT C 14. 26 patients were evaluated in this study, ages between 15-75 with upper gastrointestinal symptoms; all of them required gastroduodenal endoscopy, status H. Pylori was determined with methods upon mentioned. 24 hours after endoscopy H. Pylori antigens in stools with the technique Premier Platinum Htsa, Elisa were determined. The detection of H. Pylori antigens in stools accurately identified active H. Pylori infection. The performance characteristics of this non-invasive method was similar in sensibility and specificity to conventional tests (AU)


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Antigens, Bacterial/analysis , Feces/microbiology , Helicobacter Infections/diagnosis , Helicobacter pylori/immunology , Immunoenzyme Techniques/methods , Enzyme-Linked Immunosorbent Assay/standards , Sensitivity and Specificity
13.
Acta Gastroenterol Latinoam ; 32(2): 83-5, 2002.
Article in Spanish | MEDLINE | ID: mdl-12553159

ABSTRACT

Nowadays technics for Helicobacter pylori detection in stools like culture, and PCR, are expensive and difficult to perform. The aim of this study was to evaluate ELISA test efficacy for detection of H. Pylori antigens in stools comparing this results with standarized technics like histology (Giemsa), ureasa test and UBT C 14. 26 patients were evaluated in this study, ages between 15-75 with upper gastrointestinal symptoms; all of them required gastroduodenal endoscopy, status H. Pylori was determined with methods upon mentioned. 24 hours after endoscopy H. Pylori antigens in stools with the technique Premier Platinum Htsa, Elisa were determined. The detection of H. Pylori antigens in stools accurately identified active H. Pylori infection. The performance characteristics of this non-invasive method was similar in sensibility and specificity to conventional tests.


Subject(s)
Antigens, Bacterial/analysis , Feces/microbiology , Helicobacter Infections/diagnosis , Helicobacter pylori/immunology , Immunoenzyme Techniques/methods , Adolescent , Adult , Aged , Enzyme-Linked Immunosorbent Assay/standards , Female , Humans , Male , Middle Aged , Sensitivity and Specificity
14.
Med Pediatr Oncol ; 36(1): 104-7, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11464857

ABSTRACT

BACKGROUND: The expression of several genes is modulated during neuroblastoma differentiation. The retinoblastoma family proteins, pRb, p107 and pRb2/p130, act in the repression of proliferation genes, interacting mainly with the E2F transcription factors. PROCEDURE AND RESULTS: In this study, we found that, in neuroblastoma cell lines, pRb and p107 proteins decreased, undergoing progressive dephosphorylation, whereas pRb2/p130 increased at late stages of differentiation. B-myb expression was down-regulated in association with the up-regulation of pRb2/p130, the major partner of E2F on the E2F site of the B-myb promoter in differentiated cells. Transfection of each of the retinoblastoma family genes in neuroblastoma cells was able to induce neural differentiation, to inhibit 3H-thymidine incorporation, and to down-regulate B-myb promoter activity. CONCLUSIONS: In conclusion, our data suggest a major contribution of retinoblastoma proteins, and especially of pRb2/p130, in B-myb promoter regulation and demonstrate the induction of neural differentiation by p107 and pRb2/p130, suggesting a role of these proteins in triggering differentiation-specific genes.


Subject(s)
Cell Cycle Proteins , DNA-Binding Proteins/biosynthesis , Gene Expression Regulation, Neoplastic , Genes, myb , Neoplasm Proteins/physiology , Neuroblastoma/genetics , Nuclear Proteins/physiology , Phosphoproteins/physiology , Proteins , Retinoblastoma Protein/physiology , Trans-Activators/biosynthesis , Animals , Cell Differentiation/drug effects , Dimethyl Sulfoxide/pharmacology , E2F Transcription Factors , Genes, Reporter , Genes, Retinoblastoma , Humans , Luciferases/biosynthesis , Mice , Neoplasm Proteins/genetics , Neuroblastoma/pathology , Nuclear Proteins/genetics , Phosphoproteins/genetics , Promoter Regions, Genetic , Recombinant Fusion Proteins/biosynthesis , Retinoblastoma-Like Protein p107 , Retinoblastoma-Like Protein p130 , Transcription Factors/physiology , Transfection , Tretinoin/pharmacology , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/metabolism
15.
Med Pediatr Oncol ; 36(1): 93-6, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11464913

ABSTRACT

BACKGROUND AND PROCEDURE: Nm23 gene family has been associated with metastasis suppression and differentiation. We studied DR-nm23 during neuroblastoma cells differentiation. DR-nm23 expression increased after retinoic acid induction of differentiation in human cell lines SK-N-SH and LAN-5. RESULTS: In several cell lines, overexpression of DR-nm23 was associated with more differentiated phenotypes. SK-N-SH cells increased vimentin expression, increased deposition of collagen type IV, modulated integrin expression, and underwent growth arrest; the murine neuroblastoma cell line N1E-115 showed neurite outgrowth and a striking enhancement of beta1 integrin expression. Up-regulation of beta1 integrin was specifically responsible for the increase in the adhesion to collagen type I-coated plates. Finally, cells overexpressing DR-nm23 were unable to growth in soft agar. CONCLUSIONS: In conclusion, DR-nm23 expression is directly involved in differentiation of neuroblastoma cells, and its ability to affects the adhesion to extracellular substrates and to inhibit growth in soft agar suggests an involvement in the metastatic potential of neuroblastoma.


Subject(s)
Integrin beta1/biosynthesis , Isoenzymes/physiology , Monomeric GTP-Binding Proteins/physiology , Neoplasm Proteins/physiology , Neuroblastoma/pathology , Nucleoside-Diphosphate Kinase/physiology , Transcription Factors/physiology , Agar , Animals , Cell Adhesion , Cell Differentiation , Collagen/biosynthesis , Collagen/genetics , Culture Media , Gene Expression Regulation, Neoplastic , Humans , Integrin beta1/genetics , Intracellular Signaling Peptides and Proteins , Isoenzymes/biosynthesis , Isoenzymes/genetics , Mice , Monomeric GTP-Binding Proteins/biosynthesis , Monomeric GTP-Binding Proteins/genetics , NM23 Nucleoside Diphosphate Kinases , Neoplasm Metastasis , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Neurites/ultrastructure , Neuroblastoma/enzymology , Neuroblastoma/genetics , Nucleoside-Diphosphate Kinase/biosynthesis , Nucleoside-Diphosphate Kinase/genetics , Phenotype , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/physiology , Transcription Factors/biosynthesis , Transcription Factors/genetics , Transfection , Tumor Cells, Cultured/cytology , Tumor Cells, Cultured/metabolism , Vimentin/biosynthesis , Vimentin/genetics
16.
Cell Death Differ ; 7(9): 843-50, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11042679

ABSTRACT

DR-nm23 belongs to a gene family which includes nm23-H1, originally identified as a candidate metastasis suppressor gene. Nm23 genes are expressed in different tumor types where their levels have been alternatively associated with reduced or increased metastatic potential. Nm23-H1, -H2, DR-nm23 and nm23-H4 all possess NDP kinase activity. Overexpression of DR-nm23 inhibits differentiation and promotes apoptosis in hematopoietic cells. By contrast, it induces morphological and biochemical changes associated with neural differentiation in neuroblastoma cells. In this study, we show that mutations in the catalytic domain and in the serine 61 phosphorylation site, possibly required for protein-protein interactions, impair the ability of DR-nm23 to induce neural differentiation. Moreover, neuroblastoma cells overexpressing wild-type or mutant DR-nm23 are less sensitive to apoptosis triggered by serum withdrawal. By subcellular fractionation, wild-type and mutant DR-nm23 localize in the cytoplasm and prevalently in the mitochondrial fraction. In co-immunoprecipitation experiments, wild-type DR-nm23 binds other members of nm23 family, but mutations in the catalytic and in the RGD domains and in serine 61 inhibit the formation of hetero-multimers. Thus, the integrity of the NDP kinase activity and the presence of a serine residue in position 61 seem essential for the ability of DR-nm23 to trigger differentiation and to bind other Nm23 proteins, but not for the anti-apoptotic effect in neuroblastoma cells. These studies underline the tissue specificity of the biological effects induced by DR-nm23 expression.


Subject(s)
Apoptosis , Cell Differentiation , Monomeric GTP-Binding Proteins/metabolism , Neuroblastoma/pathology , Neurons/cytology , Nucleoside-Diphosphate Kinase/metabolism , Transcription Factors/metabolism , Animals , Catalytic Domain , Cell Differentiation/genetics , Cell Fractionation , Cell Size , Culture Media, Serum-Free , DNA, Complementary/metabolism , Genes, Reporter , Genes, myc , Immunoblotting , In Situ Nick-End Labeling , Mice , Monomeric GTP-Binding Proteins/chemistry , Monomeric GTP-Binding Proteins/genetics , Mutagenesis, Site-Directed , NM23 Nucleoside Diphosphate Kinases , Neuroblastoma/enzymology , Neuroblastoma/genetics , Neuroblastoma/metabolism , Neurons/metabolism , Nucleoside Diphosphate Kinase D , Phosphorylation , Precipitin Tests , Transcription Factors/chemistry , Transcription Factors/genetics , Transfection , Tumor Cells, Cultured
17.
Acta Gastroenterol Latinoam ; 29(4): 251-3, 1999.
Article in Spanish | MEDLINE | ID: mdl-10599400

ABSTRACT

Some papers report helicobacter pylori existence in bile from surgical specimens obtained during gallbladder or bile ducts surgery. The aim of this work was search by PCR, H. Pylori presence in bile specimens from patients suffering of gallbladder stones or by bile ducts stones. Bile samples were obtained by gallbladder punction during cholecystectomy in 26 patients, 19 of them with gallbladder stones and 7 also with gallbladder stones and bile duct stones. Age ranged from 22-69 years old, median 49.6 years old. Samples were sent to specialized biomolecular laboratory to perform PCR techniques. Two of 26 patients (7.6%) had positive reaction for the presence of DNA of H. Pylori in bile samples. Our research suggest that DNA of H. Pylori can be founded in bile samples patients with gallbladders and duct stones in Argentina.


Subject(s)
Cholelithiasis/microbiology , Helicobacter Infections/microbiology , Helicobacter pylori/isolation & purification , Adult , Aged , Bile Duct Diseases/microbiology , DNA, Bacterial/analysis , Humans , Middle Aged
18.
Cancer Res ; 59(14): 3365-8, 1999 Jul 15.
Article in English | MEDLINE | ID: mdl-10416595

ABSTRACT

The transcription factors of the Myb family are expressed in several tissues and play an important role in cell proliferation, differentiation, and survival In this study, the expression of A-myb, B-myb, and c-myb was investigated in a group of 64 neuroblastomas at different dinical stages by a sensitive reverse transcription-PCR tchnique and correlated with patients' survival. All of the myb genes were frequently expressed in neuroblastoma tumors. Interestingly, the expression of B-myb, which was detected in 33 cases, was associated with an increased risk of death (P = 0.027 in a univariate analysis), whereas there was no correlation with A-myb and c-myb expression. In addition, in a multivariate Cox regression analysis that included myb gene expression, MYCN status, age at diagnosis, and tumor staging, MYCN amplification and B-myb expression were independently associated to an increased risk (P < 0.01 and P = 0.015, respectively). In overall survival curves obtained by stratifying the neuroblastoma cases on the basis of MYCN status and B-myb expression, the group of patients without MYCN amplification and positive for B-myb expression had worse survival probability than that without MYCN amplification and nonexpressing B-myb (P < 0.01). In summary, these findings provide the first demonstration that B-myb expression can be a useful prognostic marker in human neuroblastoma. Moreover, B-myb expression has a prognostic value complementary to MYCN amplification and can identify a group of high-risk patients that would not be predicted on the basis of the MYCN status only.


Subject(s)
Cell Cycle Proteins , DNA-Binding Proteins/biosynthesis , Gene Amplification , Gene Expression Regulation, Neoplastic , Genes, myc , Neuroblastoma/genetics , Oncogenes , Trans-Activators/biosynthesis , Child , Child, Preschool , Follow-Up Studies , Humans , Infant , Infant, Newborn , Neuroblastoma/mortality , Neuroblastoma/pathology , Prognosis , Proportional Hazards Models , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , RNA, Neoplasm/biosynthesis , RNA, Neoplasm/genetics , Reverse Transcriptase Polymerase Chain Reaction , Survival Analysis
19.
Acta gastroenterol. latinoam ; 29(4): 251-3, 1999.
Article in Spanish | BINACIS | ID: bin-14377

ABSTRACT

Algunos trabajos describen la presencia de H. Pylori en muestras de bilis obtenidas durante la cirurgía por litiasis en vesícula y vías biliares. El objetivo de este trabajo, ha sido detectar la presencia del ADN del H. Pylori por medio de la Reacción en cadena de la Polimerasa (PCR) en muestras de bilis de pacientes con litasis vesicular y/o de vías biliares. Las muestras de bilis fueron obtenidas de 26 pacientes, 19 con litiasis vesicular y 7 con litiasis vesicular y coledociana, con edades comprendidas entre 22 a 69 años, media de 49,6 años, por punción de vesicular durante la colecistectomia. Las muestras fueron tratadas adecuadamente y preparadas para su investigación por PCR. 2 de 26 casos (7,6 por ciento) fueron positivos para la presencia en bilis del DNA del H. Pylori. Nuestro trabajo sugiere que el DNA del H P puede ser encontrado en muestras de bilis de pacientes portadores de litiasis biliar en la Argentina. (AU)


Subject(s)
Humans , Aged , Middle Aged , Adult , Helicobacter pylori/isolation & purification , Cholelithiasis/microbiology , Helicobacter Infections/microbiology , Polymerase Chain Reaction , Bile Duct Diseases/microbiology , Cholecystectomy , DNA, Bacterial/analysis
20.
Acta gastroenterol. latinoam ; 29(4): 251-3, 1999.
Article in Spanish | LILACS | ID: lil-248008

ABSTRACT

Algunos trabajos describen la presencia de H. Pylori en muestras de bilis obtenidas durante la cirurgía por litiasis en vesícula y vías biliares. El objetivo de este trabajo, ha sido detectar la presencia del ADN del H. Pylori por medio de la Reacción en cadena de la Polimerasa (PCR) en muestras de bilis de pacientes con litasis vesicular y/o de vías biliares. Las muestras de bilis fueron obtenidas de 26 pacientes, 19 con litiasis vesicular y 7 con litiasis vesicular y coledociana, con edades comprendidas entre 22 a 69 años, media de 49,6 años, por punción de vesicular durante la colecistectomia. Las muestras fueron tratadas adecuadamente y preparadas para su investigación por PCR. 2 de 26 casos (7,6 por ciento) fueron positivos para la presencia en bilis del DNA del H. Pylori. Nuestro trabajo sugiere que el DNA del H P puede ser encontrado en muestras de bilis de pacientes portadores de litiasis biliar en la Argentina.


Subject(s)
Humans , Aged , Middle Aged , Adult , Cholelithiasis/microbiology , Helicobacter Infections/microbiology , Helicobacter pylori/isolation & purification , Bile Duct Diseases/microbiology , Cholecystectomy , DNA, Bacterial/analysis , Polymerase Chain Reaction
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