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1.
Sci Rep ; 13(1): 18224, 2023 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-37880284

RESUMO

This study investigates the gamma radiation shielding properties of cement-ball clay matrix composites doped with micro- and nano-sized cadmium oxide (CdO) particles. The linear attenuation coefficient (LAC) was determined using a sodium iodide (NaI) detector and five radioactive point sources with energies ranging from 59.5 to 1408 keV. The LAC values obtained were compared to the XCOM database and found to be in good agreement. The composites' half-value layer (HVL), tenth value layer (TVL), mean free path (MFP), effective atomic number (Zeff), equivalent atomic number (Zeq), and absorption buildup factor (EABF) were determined. The results showed that the addition of CdO particles improved the radiation-shielding behavior of the composites and increasing the weight fraction of CdO particles increased the shielding effectiveness. The results also illustrated that when nano-sized CdO particles were compared to their micro-sized counterparts, there was a significant enhancement in radiation shielding effectiveness. For instance, a composite material composed of 50% cement, 41.7% ball clay, and 3.8% nano CdO at an energy level of 0.0595 MeV exhibited a remarkable 12.2% increase in attenuation, surpassing the performance of the micro-sized sample with an equivalent concentration. Similarly, another composite consisting of 50% cement, 33.3% ball clay, and 16.7% nano CdO demonstrated a significant 15.4% increase in attenuation at the same energy level, when compared to the micro-sized sample. The study demonstrates the potential of CdO-doped cement-ball clay matrix composites for gamma radiation shielding applications.

2.
J Infect Dis ; 223(12 Suppl 2): S236-S240, 2021 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-33330908

RESUMO

Intercellular communication and environmental sensing are most often mediated through ligand-receptor binding and signaling. This is true for both host cells and microbial cells. The ligands can be proteins (cytokines, growth factors, and peptides), modified lipids, nucleic acid derivatives and small molecules generated from metabolic pathways. These latter nonprotein metabolites play a much greater role in the overall function of mucosal immunity than previously recognized, and the list of potential immunomodulatory molecules derived from the microbiome is growing. The most well-studied microbial signals are the nonmetabolite microbe-associated molecular pattern molecules, such as lipopolysaccharide and teichoic acid, that bind to host pattern recognition receptors. Here, we will highlight the immunomodulatory activities of other microbiome-derived molecules, such as short-chain fatty acids, bile acids, uric acid, prostaglandins, histamine, catecholamines, aryl hydrocarbon receptor ligands, and 12,13-diHOME.


Assuntos
Imunidade nas Mucosas , Microbiota/imunologia , Animais , Apresentação de Antígeno , Humanos , Fatores Imunológicos/metabolismo , Inflamassomos/imunologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Transdução de Sinais , Linfócitos T Reguladores/imunologia
3.
J Family Med Prim Care ; 9(6): 3124-3130, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32984184

RESUMO

BACKGROUND: Little is recognized about factors affecting poor sleep hygiene and relations of sleep problems with the quality of life among adults. AIMS: To evaluate sleep hygiene, explore its associated factors, assess the prevalence of excessive daytime sleepiness (EDS) and insomnia, and their relations with sleep hygiene. Besides, correlate sleep problems with life quality. METHODS: A representative sample of primary healthcare population was surveyed using questionnaires comprising sociodemographic characters, personal lifestyles, validated Arabic versions of Epworth sleepiness scales, Athens insomnia scale, short-form health survey questionnaire (SF-12), and developed Arabic sleep hygiene index (SHI). RESULTS: A total of 401 adults participated in the study. The average SHI score was 17.25 ± 7.33. Poorer sleep hygiene was significantly detected in younger age, unmarried, unemployed, smokers, and energy drinks consumers (P < 0.05). Positive significant correlations were correlated with cellphone and video-gaming duration. About 56.61% and 39.90% of participants suffered insomnia symptoms and EDS, respectively. Significant poorer SHI was detected among insomnia and EDS sufferers. Negative significant correlations were observed between scores of both components of SF-12 and EDS, insomnia, and SHI. CONCLUSION: Significant negative associations were detected between SHI, EDS, insomnia, and both components of life quality. The role of sleep hygiene education programs in the promotion of sleep and quality of life need to be considered.

4.
Rev. méd. Chile ; 144(11): 1494-1496, nov. 2016. ilus
Artigo em Espanhol | LILACS | ID: biblio-845474

RESUMO

Gougerot and Carteaud confluent and reticulated papillomatosis (CARP) is an uncommon dermatosis characterized by hyperpigmented scaly macules or papillomatous papules coalescing into confluent patches or plaques centrally with a reticular pattern peripherally. We report a 28-year-old woman presenting at 16 weeks of gestation with an itchy rash that was biopsied and turned out to be consistent with CARP. Options for treatment were discussed but the woman refused to take any systemic therapy and used only moisturizers throughout her pregnancy. The rash subsided spontaneously after delivery.


Assuntos
Humanos , Feminino , Gravidez , Adulto , Papiloma/patologia , Complicações Neoplásicas na Gravidez/patologia , Neoplasias Cutâneas/patologia , Remissão Espontânea , Biópsia , Exantema/patologia
5.
Rev Med Chil ; 144(11): 1494-1496, 2016 Nov.
Artigo em Espanhol | MEDLINE | ID: mdl-28394969

RESUMO

Gougerot and Carteaud confluent and reticulated papillomatosis (CARP) is an uncommon dermatosis characterized by hyperpigmented scaly macules or papillomatous papules coalescing into confluent patches or plaques centrally with a reticular pattern peripherally. We report a 28-year-old woman presenting at 16 weeks of gestation with an itchy rash that was biopsied and turned out to be consistent with CARP. Options for treatment were discussed but the woman refused to take any systemic therapy and used only moisturizers throughout her pregnancy. The rash subsided spontaneously after delivery.


Assuntos
Papiloma/patologia , Complicações Neoplásicas na Gravidez/patologia , Neoplasias Cutâneas/patologia , Adulto , Biópsia , Exantema/patologia , Feminino , Humanos , Gravidez , Remissão Espontânea
6.
Radiat Prot Dosimetry ; 151(1): 117-28, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22155752

RESUMO

The tissue equivalent proportional counter (TEPC) that utilises a gas cavity has been the standard to obtain microdosimetric observations. An alternative is the solid-state microdosimeter that replaces the gas with a solid-state detector with microscopic sensitive volumes. Here, we describe the development of two versions of a personal solid-state microdosimeter for space exploration applications and give test results for iron and proton beams with comparisons to TEPC measurements and Geant4 radiation transport code simulations. In addition, we describe and provide test results of an optical technique to carry out an end-to-end system test and calibration of a silicon solid-state microdosimeter. This technique eliminates the need for an ionising radiation source with its attendant issues on use and transportation and provides an advantage over the TEPC.


Assuntos
Ferro , Prótons , Radiometria/instrumentação , Radiometria/métodos , Calibragem , Desenho de Equipamento , Humanos , Doses de Radiação , Proteção Radiológica , Silício
7.
Radiat Prot Dosimetry ; 143(2-4): 398-401, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21199825

RESUMO

Radiation in space generally produces higher dose rates than that on the Earth's surface, and contributions from primary galactic and solar events increase with altitude within the magnetosphere. Presently, no personnel monitor is available to astronauts for real-time monitoring of dose, radiation quality and regulatory risk. This group is developing a prototypic instrument for use in an unknown, time-varying radiation field. This microdosemeter-dosemeter nucleon instrument is for use in a spacesuit, spacecraft, remote rover and other applications. It provides absorbed dose, dose rate and dose equivalent in real time so that action can be taken to reduce exposure. Such a system has applications in health physics, anti-terrorism and radiation-hardening of electronics as well. The space system is described and results of ground-based studies are presented and compared with predictions of transport codes. An early prototype in 2007 was successfully launched, the only solid-state microdosemeter to have flown in space.


Assuntos
Materiais Biomiméticos , Carga Corporal (Radioterapia) , Radiação Cósmica , Monitoramento de Radiação/instrumentação , Astronave/instrumentação , Contagem Corporal Total/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Miniaturização , Doses de Radiação , Eficiência Biológica Relativa , Medição de Risco/métodos
8.
Radiat Prot Dosimetry ; 140(4): 309-18, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20430854

RESUMO

The gold standard in microdosemeters has been the tissue equivalent proportional counter (TEPC) that utilises a gas cavity. An alternative is the solid-state microdosemeter that replaces the gas with a condensed phase (silicon) detector with microscopic sensitive volumes. Calibrations of gas and solid-state microdosemeters are generally carried out using radiation sources built into the detector that impose restrictions on their handling, transportation and licensing in accordance with the regulations from international, national and local nuclear regulatory bodies. Here a novel method is presented for carrying out a calibration and end-to-end system test of a microdosemeter using low-energy photons as the initiating energy source, thus obviating the need for a regulated ionising radiation source. This technique may be utilised to calibrate both a solid-state microdosemeter and, with modification, a TEPC with the higher average ionisation energy of a gas.


Assuntos
Radiometria/instrumentação , Radiometria/métodos , Algoritmos , Calibragem , Elétrons , Desenho de Equipamento , Humanos , Transferência Linear de Energia , Teste de Materiais , Oscilometria/métodos , Fótons , Física/métodos , Doses de Radiação , Monitoramento de Radiação/métodos , Proteção Radiológica/métodos
9.
Saudi Med J ; 25(5): 663-6, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15138539

RESUMO

We report a patient with pulmonary alveolar microlithiasis who was admitted to King Abdul-Aziz Hospital, Makkah, Kingdom of Saudi Arabia with chest pain, shortness of breath, dry cough and swelling of lower limbs. The patient underwent chest radiographs and CT scan showing multiple diffuse, almost symmetrical bilateral micronodular opacities of calcific density. The diagnosis was confirmed after percutaneous lung biopsy from the patient. Cardiokinetics, diuretics and oxygen were administered with slight improvement.


Assuntos
Calcinose/diagnóstico , Pneumopatias/diagnóstico , Alvéolos Pulmonares , Biópsia , Calcinose/patologia , Diagnóstico Diferencial , Feminino , Humanos , Pulmão/patologia , Pneumopatias/patologia , Pessoa de Meia-Idade , Alvéolos Pulmonares/patologia , Tomografia Computadorizada por Raios X
10.
Eur J Biochem ; 267(13): 4242-52, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10866829

RESUMO

The bacterial histidine permease is a model system for ABC transporters (traffic ATPases). The water-soluble receptor of this permease, HisJ, binds L-histidine and L-arginine (tightly) and L-lysine and L-ornithine (less tightly) in the periplasm, interacts with the membrane-bound complex (HisQMP2) and induces its ATPase activity, which results in ligand translocation. HisJ is a two-domain protein; in the absence of ligand, the cleft between two domains is open and binding of substrate stabilizes the closed conformation. Surprisingly, various liganded HisJ forms display substantial differences in their physicochemical characteristics and capacity to induce the ATPase. This is due to either different effects of the individual ligands on the respective closed structures, or to different equilibria being reached for each ligand between the open liganded form and the closed liganded form [Wolf, A. , Lee, K.C., Kirsch, J.F. & Ames, G.F.-L. (1996) J. Biol. Chem. 271, 21243-21250]. In this work, time-resolved measurements of the decay of intrinsic HisJ fluorescence and of the decay of the anisotropy of the fluorescence, as well as the analysis of the steady-state near UV CD and fluorescence spectra, rule out the model in which the differences between liganded complexes reflect different equilibria. The decay of the anisotropy of the fluorescence shows that liganded complexes differ dramatically in their large-scale conformational dynamics. Differential scanning calorimetry (DSC) curves for the HisJ thermal unfolding are well described by a scheme of equilibrium two-state unfolding of two independent domains, which can be ascribed to the two-domain structure of HisJ. This is true both for apo-HisJ at various pH values, and for HisJ in the presence of its ligands at varying concentrations, at pH 8.3. The DSC and structural data suggest that all ligands interact more extensively with the larger domain. A qualitative model for the HisJ conformational dynamics employing the idea of a twisting movement of the domains is proposed, which explains the difference in the efficacy of the ATPase induction by the various liganded HisJ forms.


Assuntos
Transportadores de Cassetes de Ligação de ATP , Sistemas de Transporte de Aminoácidos Básicos , Proteínas de Bactérias , Proteínas de Transporte/química , Proteínas de Membrana Transportadoras/metabolismo , Proteínas Periplásmicas de Ligação , Termodinâmica , Adenosina Trifosfatases/metabolismo , Varredura Diferencial de Calorimetria , Dicroísmo Circular , Ligantes , Conformação Proteica , Espectrometria de Fluorescência
11.
J Biochem ; 126(3): 545-52, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10467170

RESUMO

beta-Glycosidase from the extreme thermophilic archaeon Sulfolobus solfataricus is a tetrameric protein with a molecular mass of 240 kDa, stable in the presence of detergents, and with a maximal activity at temperatures above 95 degrees C. Understanding the structure-activity relationships of the enzyme under different conditions is of fundamental importance for both theoretical and applicative purposes. In this paper we report the effect of methanol, ethanol, 1-propanol, and 1-butanol on the activity of S. solfataricus beta-glycosidase expressed in Escherichia coli. The alcohols stimulated the enzyme activity, with 1-butanol producing its maximum effect at a lower concentration than the other alcohols. The structure of the enzyme was studied in the presence of 1-butanol by circular dichroism, and Fourier-transform infrared and fluorescence spectroscopies. Circular dichroism and steady-state fluorescence measurements revealed that at low temperatures the presence of the alcohol produced no significant changes in the tertiary structure of the enzyme. However, time-resolved fluorescence data showed that the alcohol modifies the protein microenvironment, leading to a more flexible enzyme structure, which is probably responsible for the enhanced enzymatic activity.


Assuntos
1-Butanol/química , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/metabolismo , Sulfolobus/enzimologia , Dicroísmo Circular , Clonagem Molecular , Escherichia coli/genética , Cinética , Desnaturação Proteica , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Relação Estrutura-Atividade
13.
Biophys Chem ; 79(1): 25-32, 1999 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-10371019

RESUMO

We observed emission from the tyrosine derivative N-acetyl-L-tyrosinamide (NATyrA) when excited with the fundamental output of a femtosecond Ti:Sapphire laser from 780 to 855 nm. The dependence on incident laser power indicates a three-photon process. The emission spectra and intensity decay in glycerol-water (30:70) at 5 degrees C were found to be identical for one- and three-photon excitation. Also the excitation spectrum of three-photon-induced fluorescence of NATyrA corresponds to the one-photon excitation spectrum. The time-zero or fundamental anisotropy spectrum was reconstructed from the frequency-domain anisotropy decays. The three-photon anisotropies are similar or larger than the one-photon anisotropies. These three-photon anisotropies are surprising given the near zero values known for tyrosine with two-photon excitation. The observations indicate that one- and three-photon excitation directly populates the same singlet excited states(s). However, the origin of the anisotropies with multi-photon excitation of tyrosine remain unclear and unpredictable.


Assuntos
Tirosina/análogos & derivados , Polarização de Fluorescência , Técnicas In Vitro , Lasers , Fotoquímica , Fótons , Espectrometria de Fluorescência , Tirosina/química , Tirosina/efeitos da radiação
14.
Biochemistry ; 38(13): 4097-105, 1999 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-10194324

RESUMO

The bilin organization of three cryptomonad biliproteins (phycocyanins 612 and 645 and phycoerythrin 545) was examined in detail. Two others (phycocyanin 630 and phycoerythrin 566) were studied less extensively. Phycocyanin 645 and phycoerythrin 545 were suggested to have one bilin in each monomeric (alphabeta) unit of the dimer (alpha2beta2) isolated from the others, and the remaining six bilins may be in pairs. One pair was found across the monomer-monomer interface of the protein dimer, and two identical pairs were proposed to be within the monomer protein units. For phycocyanin 612, a major surprise was that a pair of bilins was apparently not found across the monomer-monomer interface, but the remaining bilins were distributed as in the other two cryptomonad proteins. The effect of temperature on the CD spectra of phycocyainin 612 demonstrated that two of the bands (one positive and one negative) behaved identically, which is required if they are coupled. The two lowest-energy CD bands of phycocyanin 612 originated from paired bilins, and the two higher-energy bands were from more isolated bilins. The paired bilins within the protein monomers contained the lowest-energy transition for these biliproteins. Using the bilins as naturally occurring reporter groups, phycocyanin 612 was shown to undergo a reversible change in tertiary structure at 40 degrees C. Protein monomers were shown to be functioning biliproteins. A hypothesis is that the coupled pair of bilins within the monomeric units offers important advantages for efficient energy migration, and other bilins transfer energy to this pair, extending the wavelength range or efficiency of light absorption.


Assuntos
Ficocianina/análogos & derivados , Ficocianina/química , Ficoeritrina/química , Pirróis/química , Dicroísmo Circular , Cianobactérias , Dimerização , Transferência de Energia , Modelos Químicos , Ficobilinas , Rodófitas , Espectrometria de Fluorescência , Temperatura , Tetrapirróis
15.
Biochemistry ; 37(19): 6636-44, 1998 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-9578547

RESUMO

Distances between the paclitaxel, colchicine, and exchangeable GTP binding sites on tubulin polymers have been probed using fluorescence spectroscopy. Techniques for measuring fluorescence resonance energy transfer (FRET) between fluorescent or chromophoric ligands for each binding site were employed. 2-Debenzoyl-2-(m-aminobenzoyl)paclitaxel (2-AB-PT) was the fluorophore ligand for the paclitaxel binding site; thiocolchicine, allocolchicine, and MDL 27048 were probes for the colchicine site, and 2'(or 3')-O-(trinitrophenyl)guanosine 5'-triphosphate (TNP-GTP) was the fluorophore ligand for the exchangeable GTP site. The distance between the colchicine and paclitaxel binding sites was determined with two different acceptor ligands in the colchicine site. An average distance distribution of 17 A was found in both cases. Energy transfer between 2-AB-PT bound to the paclitaxel site and TNP-GTP (acceptor) bound to the exchangeable GTP site was observed in the polymer. The average distance distribution between the fluorophores was 16.0 A, but the half-width of the distribution was large (17.9 A), which indicates that energy transfer between more than one donor-acceptor pair occurred in the system. One interpretation of this result is that 2-AB-PT serves as an energy transfer donor for two GTP sites, one contained on the same subunit and one on an adjacent protofilament. No FRET was observed between ligands bound to the colchicine and exchangeable GTP sites, indicating that the result of colchicine binding on the GTP region of beta-tubulin is a long range, allosteric effect. The results from these experiments are interpreted in terms of known structural features of microtubules.


Assuntos
Colchicina/química , Guanosina Trifosfato/química , Paclitaxel/química , Tubulina (Proteína)/química , Animais , Sítios de Ligação/efeitos dos fármacos , Bovinos , Colchicina/metabolismo , Transferência de Energia/efeitos dos fármacos , Guanosina Trifosfato/metabolismo , Guanosina Trifosfato/farmacologia , Ligantes , Modelos Moleculares , Paclitaxel/metabolismo , Espectrometria de Fluorescência , Trinitrobenzenos/química , Trinitrobenzenos/farmacologia , Tubulina (Proteína)/metabolismo
16.
Biochemistry ; 37(1): 417-23, 1998 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-9425063

RESUMO

Phycoerythrin 545 was isolated having an alpha2beta2 (dimer) protein structure at pH 6.0 and 2 g/L protein concentration with eight bilin chromophores. Monomers (alphabeta) were produced by lowering the protein concentration to 0.15 g/L and the pH to 4.5. Dimer dissociation was monitored by dynamic light scattering and gel-filtration column chromatography. Monomers were stable and had bilin optical spectra different from the alpha2beta2 dimers, although they have very similar protein secondary structures. The optical spectra of phycoerythrin 545 showed four types of behavior with temperature: 10-20 degrees C, dimers; 40-50 degrees C, dimers/monomers; 60 degrees C, nearly fully disordered; 70 degrees C, disordered alpha and beta polypeptides. At 40 degrees C, the protein dissociated partially to monomer, which could be totally reversed to dimers at 20-25 degrees C. The visible circular dichroism difference spectrum for the protein dimers minus monomers exhibited positive and negative bands--such spectra may indicate exciton splitting between closely-spaced bilins. Circular dichroism also revealed a spectrum suggesting exciton coupling for the second excited state of the bilins. Ultrafast fluorescence using a two-photon method showed the fastest time for protein dimers to be 2. 4 ps and monomers had a 39-ps lifetime. Phycocyanin 645 was found to have a 550-fs lifetime.


Assuntos
Pigmentos Biliares/química , Eucariotos/química , Ficoeritrina/química , Pigmentos Biliares/metabolismo , Sítios de Ligação , Cromatografia em Gel , Dicroísmo Circular , Dimerização , Transferência de Energia , Luz , Ficoeritrina/metabolismo , Espalhamento de Radiação , Espectrometria de Fluorescência , Temperatura
17.
Biophys Chem ; 67(1-3): 35-41, 1997 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-9397517

RESUMO

We examined the steady state and time-resolved emission of DNA stained with ethidium bromide (EB) when excited with 90 fs pulses from a mode-locked titanium sapphire laser. Over the wavelength range from 840 to 880 nm EB-DNA was found to display two-photon excitation, with a cross-section near 7 x 10(-50) cm4s/photon. Frequency-domain intensity decay measurements revealed similar multi-exponential intensity decays for one- and two-photon excitation. Time-resolved anisotropy decay measurements revealed similar correlation times, but different amplitudes as has been observed previously for two- versus one-photon excitation. These results indicate that two-photon excitation of EB-DNA can be accomplished with the fundamental output of a Ti:sapphire laser without obvious heating or perturbation of the DNA.


Assuntos
DNA/química , Etídio/química , Fótons , Animais , Bovinos , Polarização de Fluorescência , Espectrometria de Fluorescência/métodos
18.
Biophys J ; 72(2 Pt 1): 567-78, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9017187

RESUMO

We examined the fluorescence spectral properties of the DNA stains DAPI (4',6-diamidino-2-phenylindole, hydrochloride) and Hoechst 33342 (bis-benzimide, or 2,5'-bi'1H-benzimidazole2'-(4-ethoxyphenyl)-5-(4-methyl-1-piperazi nyl)) with two-photon (2h nu) and three-photon (3h nu) excitation using femtosecond pulses from a Ti:sapphire laser from 830 to 885 nm. The mode of excitation of DAPI bound to DNA changed from two-photon at 830 nm to three-photon at 885 nm. In contrast, Hoechst 33342 displayed only two-photon excitation from 830 to 885 nm. DAPI-DNA displayed the same emission spectra and decay times for 2h nu and 3h nu excitation. Hoechst 33342-DNA displayed the same intensity decay for excitation at 830 and 885 nm. Both probes displayed higher anisotropies for multiphoton excitation as compared to one-photon excitation with ultraviolet wavelengths, and DAPI-DNA displays a higher anisotropy for 3h nu at 885 nm than for 2h nu at 830 nm. We used 970-nm excitation of DAPI-stained chromosomes to obtain the first three-dimensional images with three-photon excitation. Three-photon excitation of DAPI-stained chromosomes at 970 nm was demonstrated by the power dependence in the fluorescence microscope.


Assuntos
Benzimidazóis , DNA/química , Corantes Fluorescentes , Indóis , Espectrometria de Fluorescência , Fenômenos Químicos , Físico-Química , Cromossomos , Polarização de Fluorescência , Células HeLa , Humanos , Processamento de Imagem Assistida por Computador , Cinética , Lasers , Luz , Metáfase , Microscopia de Fluorescência , Fótons , Espectrofotometria
19.
Biophys J ; 71(6): 3448-53, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8968614

RESUMO

We report the first measurements of protein fluorescence with three-photon excitation, using a mutant of troponin C (TnC) that contains a single tryptophan residue F22W. From the emission intensity dependence on laser power we determine that TnC F22W displays one-, two-, and three-photon excitation at 285, 570, and 855 nm, respectively. The emission spectra and intensity decays are identical for one-, two-, or three-photon excitation. The steady-state and time 0 anisotropies are distinct for each mode of excitation, but the correlation times were the same, suggesting that three-photon excitation of proteins can be accomplished without significant effects of the locally intense illumination. The excitation anisotropy spectrum from 830 to 900 nm displays only negative values, suggesting dominant excitation via the 1Lb state of tryptophan from 830 to 900 nm.


Assuntos
Conformação Proteica , Troponina C/química , Triptofano , Animais , Embrião de Galinha , Galinhas , Polarização de Fluorescência , Concentração de Íons de Hidrogênio , Cinética , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Mutagênese Sítio-Dirigida , Fótons , Mutação Puntual , Proteínas Recombinantes/química , Espectrometria de Fluorescência/métodos
20.
Anal Biochem ; 242(2): 266-70, 1996 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-8937572

RESUMO

The anticancer drug topotecan was detected in human plasma and whole blood using two-photon excitation at 730 or 820 nm. These wavelengths are longer than the main absorption bands of hemoglobin. Two-photon excitation of topotecan was demonstrated by a quadratic dependence of the emission intensity on the incident power, compared to a linear dependence for one-photon excitation at 410 nm. The observed emission centered at 525 nm was shown to be topotecan from the similarity of the emission spectrum and decay times observed for one-photon and two-photon excitation. Topotecan was detected at concentrations as low as 0.05 and 1 microM in plasma and whole blood, respectively. Since skin blood and tissues are translucent at long wavelengths, these results suggest the possibility of homogeneous or noninvasive clinical sensing with two-photon excitation.


Assuntos
Antineoplásicos/sangue , Análise Química do Sangue/métodos , Camptotecina/análogos & derivados , Espectrometria de Fluorescência/métodos , Antineoplásicos/química , Camptotecina/sangue , Camptotecina/química , Técnicas de Química Analítica/métodos , Humanos , Raios Infravermelhos , Lasers , Masculino , Fótons , Plasma/química , Pele/irrigação sanguínea , Topotecan
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