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We propose an unsupervised regularized inversion method for reconstruction of the 3D refractive index map of a sample in tomographic diffractive microscopy. It is based on the minimization of the generalized Stein's unbiased risk estimator (GSURE) to automatically estimate optimal values for the hyperparameters of one or several regularization terms (sparsity, edge-preserving smoothness, total variation). We evaluate the performance of our approach on simulated and experimental limited-view data. Our results show that GSURE is an efficient criterion to find suitable regularization weights, which is a critical task, particularly in the context of reducing the amount of required data to allow faster yet efficient acquisitions and reconstructions.
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"General-purpose cohorts" in epidemiology and public health are designed to cover a broad scope of determinants and outcomes, in order to answer several research questions, including those not defined at study inception. In this context, the general objective of the CONSTANCES project is to set up a large population-based cohort that will contribute to the development of epidemiological research by hosting ancillary projects on a wide range of scientific domains, and to provide public health information. CONSTANCES was designed as a randomly selected sample of French adults aged 18-69 years at study inception; 202,045 subjects were included over an 8-year period. At inclusion, the selected participants are invited to attend one of the 24 participating Health Prevention Centers (HPCs) for a comprehensive health examination. The follow-up includes a yearly self-administered questionnaire, and a periodic visit to an HPC. Procedures have been developed to use the national healthcare databases to allow identification and validation of diseases over the follow-up. The biological collection (serum, lithium heparinized plasma, EDTA plasma, urine and buffy coat) began gradually in June 2018. At the end of the inclusions, specimens from 83,000 donors will have been collected. Specimens are collected according to a standardized protocol, identical in all recruitment centers. All operations relating to bio-banking have been entrusted by Inserm to the Integrated Biobank of Luxembourg (IBBL). A quality management system has been put in place. Particular attention has been paid to the traceability of all operations. The nature of the biological samples stored has been deliberately limited due to the economic and organizational constraints of the inclusion centers. Some research works may require specific collection conditions, and can be developed on request for a limited number of subjects and in specially trained centers. The biological specimens that are collected will allow for a large spectrum of biomarkers studies and genetic and epigenetic markers through candidate or agnostic approaches. By linking the extensive data on personal, lifestyle, environmental, occupational and social factors with the biomarker data, the CONSTANCES cohort offers the opportunity to study the interplays between these factors using an integrative approach and state-of-the-art methods.
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Bancos de Espécimes Biológicos , Adolescente , Adulto , Idoso , Estudos de Coortes , Bases de Dados Factuais , Humanos , Luxemburgo , Pessoa de Meia-Idade , Inquéritos e Questionários , Adulto JovemRESUMO
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect often presenting with neonatal jaundice and/or hemolytic anemia. G6PD hemolytic events are linked with exposure to a pro-oxidant agent. We here report three cases of initial G6PD crises in breastfed children secondary to maternal consumption of a tonic drink which contains quinine. Quinine was found in breast milk of one of the mothers after she consumed tonic water. CONCLUSION: The amount of quinine that is transmitted through breast milk appears to be sufficient to induce G6PD crises in breastfed children. We hence recommend that consumption of quinine-containing sodas during breastfeeding should be avoided in populations with a high prevalence of G6PD deficiency. What is Known: ⢠G6PD hemolytic events are linked with exposure to a pro-oxidant agent. ⢠Ingestion of fava beans by a mother who was breastfeeding has been reported to induce a neonatal G6PD crisis. What is New: ⢠Maternal consumption of tonic drink which contains quinine appears to be sufficient to induce G6PD crises in breastfed children. ⢠Maternal consumption of quinine-containing sodas during breastfeeding should be avoided in populations with a high prevalence of G6PD deficiency.
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Aleitamento Materno , Bebidas Gaseificadas/toxicidade , Deficiência de Glucosefosfato Desidrogenase/induzido quimicamente , Oxidantes/toxicidade , Quinina/toxicidade , Feminino , Deficiência de Glucosefosfato Desidrogenase/diagnóstico , Humanos , Lactente , Recém-Nascido , MasculinoRESUMO
INTRODUCTION: Hypernatraemia in hospitalised patients is less common and less studied than hyponatraemia, although it also seems to be associated with a poor prognosis. The present study evaluates its prevalence, risk factors and prognosis in an internal medicine department. METHODS: Full hospital stays over 28 months in a 36-bed internal medicine department were analysed retrospectively. Patients with at least one plasma sodium ≥ 150 mmol/l were compared first with all other patients and then individually with sex- and age-matched normonatraemic controls. RESULTS: Plasma sodium ≥ 150 mmol÷l was observed during 49÷1945 hospitalisations (2.6%); it was acquired during hospitalisation in 30 cases (61%). Hypernatraemic patients were significantly older with no gender difference. They were comparable with their matched normonatraemic controls regarding the Charlson comorbidity index, although individual comorbidities varied. They were bedridden in 45% vs 15% for controls (p = 0.001). Nearly one-third of hypernatraemic patients had an increased extracellular fluid volume. Hypernatraemia was associated with higher in-hospital mortality (43% vs 2%, p < 0.001) and longer hospitalisation (median 21 vs 10 days, p = 0.004). CONCLUSION: Hypernatraemia is more likely to occur in older and dependent patients and is associated with poor prognosis. Unlike classical teaching, it is often associated with increased extracellular fluid volume, even outside intensive care units.
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Cardiopatias/epidemiologia , Mortalidade Hospitalar , Hospitalização , Hipernatremia/epidemiologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Estudos de Coortes , Comportamento de Ingestão de Líquido , Líquido Extracelular , Feminino , França/epidemiologia , Humanos , Medicina Interna , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Limitação da Mobilidade , Prevalência , Prognóstico , Estudos Retrospectivos , Fatores de RiscoRESUMO
Antioxidants may affect the outcome of photodynamic therapy (PDT) through the inactivation of reactive oxygen species. Their direct interaction with photosensitizers excited at the triplet state is also worthy of interest. This process is investigated by laser flash photolysis of m-THPC (meso-tetra(3-hydroxyphenyl)chlorin, Foscan) hydroalcoholic solutions added with Trolox (TrOH), a standard antioxidant or Propofol (PfOH, Diprivan(®)), a common anesthetic agent also characterized for its antioxidant properties. Transient UV-visible absorption spectra, kinetics at selected wavelengths and final spectra after extensive laser irradiation show that both compounds react with the m-THPC triplet state, (3)m-THPC, to ultimately restore the photosensitizer in its ground state. For PfOH, this process mainly appears as a single step obeying pseudo-first order kinetics. The bimolecular rate constant for the quenching of (3)m-THPC by PfOH is around 2 × 10(6) M(-1) s(-1), a value increased to some extent by the water content of the solution. A bimolecular reaction between (3)m-THPC and TrOH is observed with a rate constant of similar magnitude and dependence upon water. However, the reaction leads, at least partly, to intermediate species assigned to the TrOË radical and the m-THPC anion radical. Within a few ms, these species back react to yield m-THPC in its ground state. A general mechanism involving an intermediate activated complex with some charge transfer character is proposed. Depending on the redox potentials for the oxidation of the antioxidant, this complex evolves predominantly either toward the formation of radicals (TrOH) or back to the photosensitizer ground state (PfOH). Notably, the kinetics data suggest that Propofol may quench (3)m-THPC at concentrations relevant of clinical situation in PDT involving anesthesia.
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Anestésicos/química , Antioxidantes/química , Cromanos/química , Mesoporfirinas/química , Fármacos Fotossensibilizantes/química , Propofol/química , Anestésicos/uso terapêutico , Antioxidantes/uso terapêutico , Cromanos/uso terapêutico , Humanos , Cinética , Lasers , Mesoporfirinas/uso terapêutico , Neoplasias/tratamento farmacológico , Oxirredução , Fotoquimioterapia , Fotólise , Fármacos Fotossensibilizantes/uso terapêutico , Propofol/uso terapêutico , Espécies Reativas de Oxigênio/metabolismoRESUMO
Photodynamic therapy (PDT) is a cancer treatment modality involving the combination of light, a photosensitizer (PS) and molecular oxygen, which results in the production of cytotoxic reactive oxygen species (ROS). Singlet oxygen ((1)O(2)) is one of the most important of these ROS. Because the lifetime and diffusion of (1)O(2) is very limited, a controllable singlet oxygen generation with high selectivity and localization would lead to more efficient and reliable PDT. The lack of selective accumulation of the PS within tumour tissue is a major problem in PDT. Targeted PDT would offer the advantage to enhance photodynamic efficiency by directly targeting diseased cells or tissues. Many attempts have been made to either selectively deliver light to diseased tissues or increase the uptake of the photoactive compounds by the target cells. The review will survey the literature regarding the multi-level control of (1)O(2) production for PDT applications. The mechanisms of ROS formation are described. The different strategies leading to targeted formation of (1)O(2) are developed. Some active PDT agents have been based on energy transfer between PS by control of the aggregation/ disaggregation. The concept of molecular beacon based on quenching-dequenching upon protease cleavage is capable of precise control of (1)O(2) by responding to specific cancer-associated biomarkers.
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Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Humanos , Neoplasias/tratamento farmacológico , Peptídeo Hidrolases/metabolismo , Fármacos Fotossensibilizantes/uso terapêutico , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Espécies Reativas de Oxigênio/toxicidadeRESUMO
We have assessed the possibility to build a prognosis predictor (PP), based on non-neoplastic mucosa microarray gene expression measures, for stage II colon cancer patients. Non-neoplastic colonic mucosa mRNA samples from 24 patients (10 with a metachronous metastasis, 14 with no recurrence) were profiled using the Affymetrix HGU133A GeneChip. Patients were repeatedly and randomly divided into 1000 training sets (TSs) of size 16 and validation sets (VS) of size 8. For each TS/VS split, a 70-gene PP, identified on the TS by selecting the 70 most differentially expressed genes and applying diagonal linear discriminant analysis, was used to predict the prognoses of VS patients. Mean prognosis prediction performances of the 70-gene PP were 81.8% for accuracy, 73.0% for sensitivity and 87.1% for specificity. Informative genes suggested branching signal-transduction pathways with possible extensive networks between individual pathways. They also included genes coding for proteins involved in immune surveillance. In conclusion, our study suggests that one can build an accurate PP for stage II colon cancer patients, based on non-neoplastic mucosa microarray gene expression measures.
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Neoplasias do Colo/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise por Conglomerados , Neoplasias do Colo/metabolismo , Feminino , Humanos , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Mucosa/metabolismo , Mucosa/patologia , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Prognóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e EspecificidadeRESUMO
Singlet oxygen (1O2), a reactive oxygen species, has been found to be implicated in many cellular events and pathological disorders. Herein, we investigated the reactivity of 1O2 towards the anaesthetic agent propofol (PPF) encapsulated within DMPC liposomes. By time resolved luminescence, the rate constant of 1O2 quenching by PPF was evaluated, depending on the location of the sensitizer, with following values: 1.35+/-0.05x10(7) M(-1) s(-1) for deuteroporphyrin (as embedded source) and 0.8+/-0.04x10(7) M(-1) s(-1) for uroporphyrin (as external source), respectively. The nature of the oxidation product, resulting from the reaction of 1O2 with PPF, was determined using absorption and HPLC techniques. Finally, the in vitro protective effect of PPF towards the 1O2-induced neuronal cell toxicity was evaluated in terms of cell viability.
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Anestésicos Intravenosos/farmacologia , Sequestradores de Radicais Livres/farmacologia , Neurônios/efeitos dos fármacos , Propofol/farmacologia , Oxigênio Singlete/antagonistas & inibidores , Anestésicos Intravenosos/química , Animais , Cápsulas/química , Células Cultivadas , Dimiristoilfosfatidilcolina/química , Sequestradores de Radicais Livres/química , Lipossomos/química , Camundongos , Oxirredução , Propofol/química , Oxigênio Singlete/toxicidadeRESUMO
INTRODUCTION: Ophthalmologists contend with a wide range of painful acute and chronic diseases. However, there is no tool specific to ocular pain that aids the patient in describing and quantifying pain. PURPOSE: Our objective was to develop a tool that would allow the ophthalmologist to identify the patient's pain quickly and precisely in order to measure its intensity and to determine possible causes. METHODS: An interview guide was elaborated after a literature review. Structured interviews were conducted in hospitals by a clinical research associate with patients suffering from painful acute or chronic pathologies. Different types of quantification and description of pain were proposed to patients. A questionnaire was developed and tested. After the analysis of the tests, the ODEON pilot questionnaire (Objectif Douleur En Ophtalmologie et Neuro-ophtalmologie: target: ophthalmic and neuro-ophthalmic pain) was developed. RESULTS: Twenty patients presenting ten different diagnoses were interviewed. Patients preferred to quantify pain with visual analogic or graduated scales. They appreciated the help of pictograms to describe their pain. Eight other patients presenting six different diagnoses tested the questionnaire. They judged the test version valid and easy to use, except for the section on emotional descriptors. An average of approximately 20 minutes was necessary to complete the questionnaire. After the tests, various questions were combined, reformulated, deleted or added. The ODEON pilot questionnaire contains five sections: 1. general health, 2. eyes and eyesight, 3. pain, 4. pain relief, 5. pictograms and sensorial descriptors. The closed- and open-ended questions included in these dimensions make it possible to measure patient pain and help the practitioner with patient management. CONCLUSION: The ODEON pilot questionnaire was developed under the supervision of a pilot committee involving clinicians and methodologists. Patients have indicated acceptance of this self-administered questionnaire during the cognitive debriefing and it is now being validated.
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Oftalmopatias/complicações , Doenças do Nervo Óptico/complicações , Medição da Dor/métodos , Dor/diagnóstico , Inquéritos e Questionários , Humanos , Dor/etiologia , Projetos PilotoRESUMO
Application fields of RT-PCR (reverse transcription-polymerase chain reaction) in clinical diagnosis comprises the assessment of viral load for RNA viruses and the analysis of gene transcription products. RT-PCR is also helpful when large genes have to be sequenced. Developments of quantitative approaches using real-time PCR recently led to a major widening of RT-PCR applications in clinical diagnosis. However, RT reaction is delicate due to its lack of reproducibility and to RNA lability and frequent contamination by DNA. In some cases additional difficulties come from the need to obtain a specific amplification in the presence of homologous sequences which might be present in higher amounts than the sequence of interest. These caveats have to be taken into account, when designing the RT protocol, and when choosing PCR primers and internal and/or external references. This review is aimed at helping the experimental setup of a RT-PCR based assay according to the objectives.
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Medicina Clínica/métodos , Técnicas e Procedimentos Diagnósticos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , HumanosRESUMO
The reaction of singlet oxygen with four vinyl-substituted dicarboxylic porphyrins, vinyldeuteroporphyrin (VD), ethylvinyldeuteroporphyrin (EVD), hydroxyethylvinyldeuteroporphyrin (HVD) and protoporphyrin (PP) in organic solutions is investigated. The main products, the "hydroxyaldehyde" chlorin-type derivatives, are formed with a concentration-dependent photochemical quantum yield that reaches a maximum of 7.4 +/- 1.6 x 10(-3). However, owing to the high turnover of singlet-oxygen production, these chlorin-type compounds are easily prepared photochemically with a chemical yield of 70% and little side product formation. In chemical ionization mass spectrometry, these compounds display an unusual fragmentation with a loss of 16 mass units. This is attributed to the loss of the oxygen bound to the saturated carbon of the modified pyrrole unit. All these compounds sensitize the formation of singlet oxygen with a yield around 0.8. They interact with singlet oxygen with rate constants of 5 x 10(6)-9 x 10(6) M-1 s-1, lower than those measured for vinyl porphyrins. These data are likely to help in the characterization of photoproducts of vinyl porphyrins relevant to photodynamic therapy (PP, HVD). As exemplified with VD and EVD, they also point out the reaction of singlet oxygen as an efficient route to chlorin-type photosensitizers.
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Fármacos Fotossensibilizantes/química , Porfirinas/química , Compostos de Vinila/química , Cromatografia Líquida de Alta Pressão , Deuteroporfirinas/química , Deuteroporfirinas/efeitos da radiação , Isomerismo , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Oxigênio/análise , Oxigênio/química , Fotoquímica , Fármacos Fotossensibilizantes/efeitos da radiação , Porfirinas/efeitos da radiação , Protoporfirinas/química , Protoporfirinas/efeitos da radiação , Oxigênio Singlete , Espectrofotometria , Relação Estrutura-Atividade , Compostos de Vinila/efeitos da radiaçãoRESUMO
Photodynamic therapy (PDT) using meta-tetrahydroxyphenylchlorin (mTHPC) performed on HT29 human colon adenocarcinoma xenografts in nude mice was shown to be enhanced by Trolox, a water-soluble vitamin E analogue. Trolox, injected i.p. at 250 mg/kg body weight 90 min before PDT, delayed tumor doubling time from 13 (PDT only) to 19 days. Enhancement of the tumoricidal effect of PDT by Trolox required the presence of the drug at the photochemical stage since its injection after irradiation is ineffective. HPLC measurements indicated that 1 hr after injection the Trolox concentration in plasma was as high as 0.8 mM. In vivo measurements of mTHPC fluorescence in mice treated by PDT with or without Trolox injection showed that Trolox did not protect mTHPC from photodegradation. Laser flash photolysis studies performed in solution demonstrated that Trolox reduces triplet mTHPC efficiently (reaction rate constant 2 x 10(7) M(-1) * sec(-1)) leading to the formation of radical products. Kinetic considerations suggest that the Trolox-mediated radical pathway can work in relay with singlet oxygen in hypoxic conditions, providing a possible explanation for the observed enhancement of mTHPC-sensitized PDT by Trolox.
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Antineoplásicos/uso terapêutico , Cromanos/farmacologia , Mesoporfirinas/uso terapêutico , Neoplasias Experimentais/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Cromanos/uso terapêutico , Modelos Animais de Doenças , Interações Medicamentosas , Feminino , Células HT29 , Humanos , Mesoporfirinas/farmacocinética , Camundongos , Camundongos Nus , Transplante de Neoplasias , Neoplasias Experimentais/metabolismo , Oxigênio/metabolismo , Fármacos Fotossensibilizantes/farmacocinética , Transplante HeterólogoRESUMO
5,9-Dimethyldibenzo[c,g]carbazole (DMDBC), a potent mouse hepatocarcinogen, has been shown to induce a non-linear increase in mutant frequency in the liver of the transgenic MutaMouse. To gain insight into the mechanisms underlying the mutagenicity of DMDBC in vivo, DNA damage formation and removal were monitored in mouse hepatocytes over 4-144 h after a single skin application of 10 or 90 mg/kg DMDBC. DNA adducts were measured by (32)P-post-labeling. DNA repair was assessed by: (i) the unscheduled DNA synthesis (UDS) assay, which measures [(3)H]thymidine incorporation into hepatocyte DNA undergoing excision repair; (ii) the Comet assay, which detects DNA strand breaks transiently produced between the incision and rejoining steps of the excision repair process. A plateau of approximately 400 DNA adducts/10(8) nucleotides was reached 24 h after treatment with 10 mg/kg and remained unchanged until 144 h. UDS activity was significantly induced at 15 and 24 h, while no DNA strand breaks were observed at any sampling time. These results suggest that DNA repair mechanisms were efficiently induced and the formation of a high degree of DNA damage was avoided at this dose level. Following exposure to 90 mg/kg DMDBC, the number of DNA adducts increased sharply to a maximum at 24 h ( approximately 8000/10(8) nucleotides) and then declined to approximately 500/10(8) nucleotides at 144 h. UDS activity was markedly induced from 15 to 72 h. Low levels of DNA strand breaks were observed at 24 and 48 h. The formation of large numbers of DNA adducts and the emergence of DNA strand breaks despite a strong initial induction of UDS activity suggested that DNA repair mechanisms were saturated at this dose level. This phenomenon could partly account for the non-linear induction of gene mutations previously reported in the liver of the transgenic MutaMouse.
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Carbazóis/toxicidade , Carcinógenos Ambientais/toxicidade , Adutos de DNA/metabolismo , Dano ao DNA , Fígado/efeitos dos fármacos , Animais , Reparo do DNA , Replicação do DNA , Cinética , Fígado/citologia , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Camundongos Transgênicos , Reprodutibilidade dos TestesRESUMO
beta-Propiolactone (BPL) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are two direct alkylating agents that induce multiple genetic lesions and tumors in the rodent stomach. We measured the kinetics of the induction of DNA damage by using the single-cell gel electrophoresis assay (SCGE) and the induction of gene mutations by using the MutaMouse model in the glandular stomach mucosa of mice exposed to a single oral administration of BPL or MNNG. The aims were to determine the optimal sampling time and to investigate the cause-effect relationship between DNA damage and gene mutations. The induction of comets, evaluated in individual cells with the tail moment, was analyzed 1, 2, 4, 24, and 72 hr after a single oral administration of 25 mg/kg BPL or 20 mg/kg MNNG. The effects of both compounds were most intense at the earlier sampling times (1-2 hr), tailing off 4 hr after treatment and becoming undetectable at 72 hr. The lacZ mutant frequency (MF) was measured 3, 7, 14, 28, and 50 days after a single oral administration of 150 mg/kg BPL or 100 mg/kg MNNG, and 3 and 14 days after a single administration of 25 mg/kg BPL or 20 mg/kg MNNG. The MF was strongly enhanced at the highest doses and all sampling times, the most marked effects being observed 14 days (11.1-fold) and 28 days (19.0-fold) after BPL and MNNG administration, respectively. At the lowest doses, only a small increase in MF ( approximately 2.5- to 3.5-fold) was found at both sampling times. Primary DNA damage detected with SCGE shortly after treatment (1-2 hr) was rapidly (3 days) transformed into stable gene mutations that remained detectable for 50 days. These results illustrate the ability and complementarity of the SCGE and MutaMouse models to assess the genotoxicity of direct alkylating agents in the mouse gastric mucosa in vivo.
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Dano ao DNA , Mucosa Gástrica/efeitos dos fármacos , Óperon Lac , Metilnitronitrosoguanidina/farmacologia , Mutação , Propiolactona/farmacologia , Animais , Eletroforese/métodos , Cinética , Camundongos , Camundongos Transgênicos , Mutagênicos/farmacologiaRESUMO
The interactions of cis-di-sulfonated aluminum phthalocyanine (PcS(2)Al) with dimyristoylphosphatidylcholine (DMPC) unilamellar vesicles have been investigated by fluorescence spectroscopy. At pH 7.0, PcS(2)Al incorporates into the vesicles with a high affinity constant (2.7x10(6) M(-1), in terms of phospholipid concentration). The fluorescence changes following rapid mixing of PcS(2)Al with vesicles are biphasic. The first phase is attributed to the entry of PcS(2)Al into the vesicles, as deduced from the linear dependence of the rate upon lipid concentration. More surprisingly, this rate is strongly pH dependent with a marked maximum around pH 7.3, a result interpreted in terms of the coordination state of the aluminum ion in aqueous solutions. At this pH, a hydroxide ion neutralizes the residual positive charge of the metal ion that remains unbalanced after coordination by the phthalocyanine cycle. A water molecule is likely to complete the metal coordination sphere. Only this form, PcAl(+)(OH(-))(OH(2)), with an uncharged core is quickly incorporated into the vesicles. The protonation of OH(-) or the deprotonation of the coordinated H(2)O leading to a positively or negatively charged core, respectively, account for the observed pH effect. Studies on the effect of cholesterol addition and exchange of PcS(2)Al between vesicles and albumin all indicate the absence of transfer of the phthalocyanine between the vesicle hemileaflets, a result expected from the presence of the two negatively charged sulfonated groups at the ring periphery. Instead, the slower kinetic phase is likely due to the movement of the phthalocyanine becoming more buried within the outer leaflet upon the loss of the water molecule coordinated to the aluminum ion.
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Indóis/administração & dosagem , Indóis/farmacocinética , Compostos Organometálicos/administração & dosagem , Compostos Organometálicos/farmacocinética , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/farmacocinética , Transporte Biológico Ativo , Dimiristoilfosfatidilcolina , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Indóis/química , Cinética , Lipossomos , Compostos Organometálicos/química , Fármacos Fotossensibilizantes/química , Espectrometria de FluorescênciaRESUMO
The purpose of this work was to investigate the impact of cell proliferation on liver mutagenesis. The genotoxic hepatocarcinogen 5, 9-dimethyldibenzo[c,g]carbazole (DMDBC) was administered to lacZ transgenic MutaTMMice at a non-hepatotoxic dose of 10 mg/kg, which induces only a slight increase in the liver lacZ mutant frequency (MF). To determine if cell proliferation stimuli enhanced DMDBC mutagenicity, MF was analyzed in mice first receiving DMDBC 10 mg/kg, then approximately 2 weeks later, either carbon tetrachloride (CCl4, a cytotoxic agent inducing regenerative cell proliferation) or phenobarbital (PB, a mitogenic agent inducing direct hyperplasia). In preliminary studies, the extent of cell proliferation induced by CCl4, PB and DMDBC was determined in non-transgenic CD2F1 mice by means of 5-bromodeoxyuridine labeling. The labeling index was significantly increased after CCl4 and PB, while no change was detected with DMDBC. MF was then determined in MutaTMMice 28 days after initial DMDBC treatment. No increase in MF was detected in mice receiving CCl4 or PB alone. A 2- to 3-fold increase in MF was detected in mice treated with 10 mg/kg DMDBC alone. In contrast, MF was markedly increased in mice receiving DMDBC followed by proliferative treatment (15-fold with CCl4 and 25-fold with PB). These results demonstrate that expression of DMDBC-induced mutations in mouse liver largely depends on the induction of cell proliferation (by a cytotoxic or mitogenic stimulus) and illustrate that MutaTMMouse is a valuable tool to investigate the early events of liver carcinogenesis.
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Carbazóis/farmacologia , Óperon Lac , Fígado/citologia , Mitógenos/farmacologia , Animais , Tetracloreto de Carbono/farmacologia , Divisão Celular/efeitos dos fármacos , Análise Mutacional de DNA , Frequência do Gene , Fígado/anatomia & histologia , Fígado/química , Fígado/efeitos dos fármacos , Regeneração Hepática/efeitos dos fármacos , Regeneração Hepática/genética , Masculino , Camundongos , Camundongos Transgênicos , Testes de Mutagenicidade , Mutação , Tamanho do Órgão/efeitos dos fármacos , Fenobarbital/farmacologiaRESUMO
5,9-Dimethyldibenzo[c,g]carbazole (DMDBC) is a synthetic derivative of the environmental pollutant 7H-dibenzo[c,g]carbazole. DMDBC is a potent genotoxic carcinogen specific for mouse liver. Using the MutaMouse lacZ transgenic mouse model and a positive selection assay, we measured lacZ mutant frequency (MF) in the liver 28 days after a single s.c. administration of DMDBC at 3, 10, 30, 90 or 180 mg/kg. MF remained low at 3 and 10 mg/kg, but increased markedly from 30 mg/kg onwards. To investigate the reason for this non-linear response, we examined mechanisms potentially involved in mutation induction in the liver. Genotoxic effects such as DNA adduct formation were detected in 32P-post-labelling studies. Liver sections were examined for microscopic changes and cell proliferation. These parameters, and MF, were studied 2, 4, 7, 14, 21 and 28 days after a single s.c. administration of 10 or 90 mg/kg DMDBC. At 10 mg/kg, a dose found to double the MF on day 28, DNA adducts reached a level of 200-600 adducts per 10(8) nucleotides from day 4 to day 28. No changes in histology or cell proliferation were detected at this low dose. At 90 mg/kg, MF increased gradually from day 7 to day 28 (maximum 44-fold). The DNA adduct level ranged from 400 to 4500 adducts per 10(8) nucleotides on day 2, then stabilized at approximately 400 adducts per 10(8) nucleotides on day 4. An early cytotoxic effect was detected microscopically in centrilobular hepatocytes, and was followed by liver cell proliferation. These data suggest that the marked increase in MF in MutaMouse liver after treatment in vivo with DMDBC at 90 mg/kg may be explained by the induction of replicative DNA synthesis due to a cytotoxic effect, allowing the fixation of persistent DNA adducts into mutations.
Assuntos
Carbazóis/toxicidade , Carcinógenos/toxicidade , Adutos de DNA , Óperon Lac/efeitos dos fármacos , Fígado/efeitos dos fármacos , Mutagênese , Animais , Carbazóis/farmacologia , Carcinógenos/farmacologia , Divisão Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Camundongos TransgênicosRESUMO
Some guanine-rich oligonucleotides inhibit HIV infectivity through interaction with the gp120 glycoprotein. Besides, photoinactivation of viruses attracts attention for blood decontamination. The feasibility of targeting a red light-absorbing chlorin-type photosensitizer to gp120 through covalent coupling with 8-mer phosphodiester oligodeoxynucleotides is investigated. Some conjugates inhibit binding of antibodies directed to gp120. Inhibition is significantly increased upon red light activation. The activity of the conjugates correlates with their ability to self-associate, a process strongly favored by the propensity of the hydrophobic chlorin moiety to dimerize. Thus, the photosensitizer moiety both promotes structures with a higher affinity for gp120 and, upon light activation, can induce site-directed damages to the protein.
Assuntos
Proteína gp120 do Envelope de HIV/metabolismo , Oligonucleotídeos/farmacologia , Transtornos de Fotossensibilidade , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Sequência de Aminoácidos , Dimerização , Relação Dose-Resposta à Radiação , Ensaio de Imunoadsorção Enzimática , Proteína gp120 do Envelope de HIV/efeitos dos fármacos , Proteína gp120 do Envelope de HIV/imunologia , Proteína gp120 do Envelope de HIV/efeitos da radiação , Luz , Dados de Sequência Molecular , Oligonucleotídeos/metabolismo , Porfirinas/fisiologiaRESUMO
The transfer of a dicarboxylic porphyrin from phosphatidylcholine fluid-phase unilamellar vesicles towards albumin is studied focusing on bilayer thickness and pH effects. The kinetics of this process yield the rate constants for the porphyrin flip-flop from the inner to the outer hemileaflet and its exit towards aqueous medium. Phospholipids with monounsaturated 14-22 carbon chains are used. Interplay between bilayer thickness and pH for the control of the rate constants is observed. This results in the amplification, at physiological pH, of the effect of membrane thickness on the flip-flop and exit rates as compared to pH 8.5 and 6.5. These data are explained by the degree of porphyrin burying within the bilayer resulting from a compromise between favorable hydrophobic interactions with the hydrocarbon phase and unfavorable penetration of the polar carboxylic chains. The balance between the two effects depends particularly on the neutralization of one carboxylic chain. Considering the bilayer hydrophobicity profile and the porphyrin size, the optimization of hydrophobic interactions appears dependent on the bilayer thickness. The flip-flop and the exit are governed by neutralization and deprotonation of the carboxylic chains, respectively, the rate of these proton exchanges being dependent on the porphyrin location within the bilayer.