Biomedical subjects

S L Jacques

Publications and source records attributed to S L Jacques.

At least 19 recordsLinked to original sources

PubMed2002

Selective photothermal interaction using an 805-nm diode laser and indocyanine green in gel phantom and chicken breast tissue.

Laser immunotherapy, a novel therapy for breast cancer, utilises selective photothermal interaction to raise the temperature of tumour tissue above the cell damage threshold. Photothermal interaction is achieved with intratumoral injection of a laser-absorbing dye followed by non-invasive laser irradiation. When tumour heating is used in combination with immunoadjuvant to stimulate an immune response, antitumour immunity can be achieved. In this study, the selective photothermal effect was investigated using gel phantom and chicken breast tissue. An 805-nm diode laser and indocyanine green (ICG) were used. An ICG-containing gelatin phantom was constructed to simulate targeted tumour tissue. The target gel was buried inside chicken breast tissue and the tissue-gel construct was irradiated by the laser. Temperatures at different locations in the construct were measured during the laser irradiation. For comparison, the thermal effect of an Nd:YAG laser on the tissue-gel construct was also investigated. Selective heating of target gel containing 0.27% ICG and buried 1 cm below the chicken tissue surface was achieved with the 805-nm diode laser using a power of 0.85 W and beam radius of 1 cm. The target gel experienced a temperature increase of more then 6 degrees C whereas the surrounding chicken breast tissue experienced only a minor temperature increase. The feasibility of this experimental set-up has been shown. It will be used in the future to optimise treatment parameters such as laser power, laser beam radius, and dye concentration.

Animals↗

PubMed2002

Iterative reconstruction algorithm for optoacoustic imaging.

Optoacoustic imaging is based on the generation of thermoelastic stress waves by heating an object in an optically heterogeneous medium with a short laser pulse. The stress waves contain information about the distribution of structures with preferential optical absorption. Detection of the waves with an array of broadband ultrasound detectors at the surface of the medium and applying a backprojection algorithm is used to create a map of absorbed energy inside the medium. With conventional reconstruction methods a large number of detector elements and filtering of the signals are necessary to reduce backprojection artifacts. As an alternative this study proposes an iterative procedure. The algorithm is designed to minimize the error between measured signals and signals calculated from the reconstructed image. In experiments using broadband optical ultrasound detectors and in simulations the algorithm was used to obtain three-dimensional images of multiple optoacoustic sources. With signals from a planar array of 3x3 detector elements a significant improvement was observed after about 10 iterations compared to the simple radial backprojection. Compared to conventional methods using filtered backprojection, the iterative method is computationally more intensive but requires less time and instrumentation for signal acquisition.

Journal Article↗

PubMed2001

Homoserine dehydrogenase from Saccharomyces cerevisiae: kinetic mechanism and stereochemistry of hydride transfer.

Homoserine dehydrogenase (HSD), which is required for the synthesis of threonine, isoleucine and methionine in fungi, is a potential target for novel antifungal drugs. In order to design effective inhibitors, the kinetic mechanism of Saccharomyces cerevisiae HSD and the stereochemistry of hydride transfer were examined. Product inhibition experiments revealed that yeast HSD follows an ordered Bi Bi kinetic mechanism, where NAD(P)H must bind the enzyme prior to aspartate semialdehyde (ASA) and homoserine is released first followed by NAD(P)+. H-(1,2,4-triazol-3-yl)-D,L-alanine was an uncompetitive inhibitor of HSD with respect to NADPH (K(ii)=3.04+/-0.18 mM) and a noncompetitive inhibitor with respect to ASA (K(is)=1.64+/-0.36 mM, K(ii)=3.84+/-0.46 mM), in agreement with the proposed substrate order. Both kinetic isotope and viscosity experiments provided evidence for a very rapid catalytic step and suggest nicotinamide release to be primarily rate limiting. Incubation of HSD with stereospecifically deuterated NADP[2H] and subsaturating amounts of aspartate semialdehyde revealed that the pro-S NADPH hydride is transferred to the aldehyde. The pH dependence of steady state kinetic parameters indicate that ionizable groups with basic pKs may be involved in substrate binding, consistent with the observation of Lys223 at the enzyme active site in the recently determined 3D structure [B. DeLaBarre, P.R. Thompson, G.D. Wright, A.M. Berghuis, Nat. Struct. Biol. 7 (2000) 238-244]. These findings provide the requisite foundation for future exploitation of fungal HSD in inhibitor design.

Homoserine Dehydrogenase↗

PubMed2001

Characterization of yeast homoserine dehydrogenase, an antifungal target: the invariant histidine 309 is important for enzyme integrity.

Fungal homoserine dehydrogenase (HSD) is required for the biosynthesis of threonine, isoleucine and methionine from aspartic acid, and is a target for antifungal agents. HSD from the yeast Saccharomyces cerevisiae was overproduced in Escherichia coli and 25 mg of soluble dimeric enzyme was purified per liter of cell culture in two steps. HSD efficiently reduces aspartate semialdehyde to homoserine (Hse) using either NADH or NADPH with kcat/Km in the order of 10(6-7) M(-1) x s(-1) at pH 7.5. The rate constant of the reverse direction (Hse oxidation) was also significant at pH 9.0 (kcat/Km approximately 10(4-5) M(-1) x s(-1)) but was minimal at pH 7.5. Chemical modification of HSD with diethyl pyrocarbonate (DEPC) resulted in a loss of activity that could be obviated by the presence of substrates. UV difference spectra revealed an increase in absorbance at 240 nm for DEPC-modified HSD consistent with the modification of two histidines (His) per subunit. Amino acid sequence alignment of HSD illustrated the conservation of two His residues among HSDs. These residues, His79 and His309, were substituted to alanine (Ala) using site directed mutagenesis. HSD H79A had similar steady state kinetics to wild type, while kcat/Km for HSD H309A decreased by almost two orders of magnitude. The recent determination of the X-ray structure of HSD revealed that His309 is located at the dimer interface [B. DeLaBarre, P.R. Thompson, G.D. Wright, A.M. Berghuis, Nat. Struct. Biol. 7 (2000) 238-244]. The His309Ala mutant enzyme was found in very high molecular weight complexes rather than the expected dimer by analytical gel filtration chromatography analysis. Thus the invariant His309 plays a structural rather than catalytic role in these enzymes.

Amino Acid Sequence↗

PubMed2000

Substrate-assisted catalysis of the PAR1 thrombin receptor. Enhancement of macromolecular association and cleavage.

Platelet activation and aggregation are mediated by thrombin cleavage of the exodomain of the PAR1 receptor. The specificity of thrombin for PAR1 is enhanced by binding to a hirudin-like region (Hir) located in the receptor exodomain. Here, we examine the mechanism of thrombin-PAR1 recognition and cleavage by steady-state kinetic measurements using soluble PAR1 N-terminal exodomains. We determined that the primary role of the PAR1 Hir sequence is to reduce the kinetic barriers to formation of the docked thrombin-PAR1 complex rather than to form high affinity ground-state interactions. In addition, the exosite I-bound Hir motif facilitates the productive interaction of the PAR1 (38)LDPR/SFL(44) sequence with the active site of thrombin. This locking process is the most energetically unfavorable step of the overall reaction. The subsequent irreversible steps of peptide bond cleavage are rapid and allosterically enhanced by the presence of the docked Hir sequence. Furthermore, the C-terminal exodomain product of thrombin cleavage, corresponding to the activated receptor, binds tightly to thrombin. This would suggest that an additional role of the Hir sequence in the thrombin-activated receptor is to sequester thrombin to the platelet surface and modulate cleavage of other platelet receptors such as the PAR4 thrombin receptor, which lacks a functional Hir sequence.

Amino Acid Sequence↗

PubMed2000

Imaging superficial tissues with polarized light.

OBJECTIVE: Polarized light can be used to obtain images of superficial tissue layers such as skin, and some example images are presented. This study presents a study of the transition of linearly polarized light into randomly polarized light during light propagation through tissues. STUDY DESIGN/MATERIALS AND METHODS: The transition of polarization was studied in polystyrene microsphere solutions, in chicken muscle (breast) and liver, and in porcine muscle and skin. The transition is discussed in terms of a diffusion process characterized by an angular diffusivity (radians(2)/mean free path) for the change in angular orientation of linearly polarized light per unit optical path traveled by the light. RESULTS: Microsphere diffusivity increased from 0.031 to 0.800 for diameters decreasing from 6.04 microm to 0.306 microm, respectively. Tissue diffusivity varied from a very low value (0.0004) for chicken liver to an intermediate value (0.055) for chicken and porcine muscle to a very high value (0.78) for pig skin. CONCLUSION: The results are consistent with the hypothesis that birefringent tissues randomize linearly polarized light more rapidly than nonbirefringent tissues. The results suggest that polarized light imaging of skin yields images based only on photons backscattered from the superficial epidermal and initial papillary dermis because the birefringent dermal collagen rapidly randomizes polarized light. This anatomic region of the skin is where cancer commonly arises.

Humans↗

PubMed2000

A study of aminolevulinic acid-induced protoporphyrin IX fluorescence kinetics in the canine oral cavity.

BACKGROUND AND OBJECTIVE: 5-Aminolevulinic acid-induced protoporphyrin IX is a promising photosensitizer that could enhance the spectroscopic contrast between normal and diseased oral tissues. Knowledge of the pharmacokinetics and effects on tissue type are important for diagnostic and therapeutic procedures. STUDY DESIGN/MATERIALS AND METHODS: Dogs randomly were administered five doses of 5-aminolevulinic acid: 5, 25, 50, 75, and 100 mg/kg. The fluorescence was recorded from buccal mucosa, gums, tongue, and facial skin using a fiberoptic probe connected to an optical multichannel analyzer. Blood samples were collected for hematologic and serum biochemical analysis. Pharmacokinetic parameters of interest were estimated using a compartmental model. RESULTS: Protoporphyrin fluorescence at all sites reached a peak in 2-6 hours, and returned to baseline in 24-31 hours, depending on the dose. Plasma protoporphyrin peaked earlier than oral tissues. CONCLUSION: The rate of synthesis of protoporphyrin, and its conversion to heme products are dose dependent. Different tissues have different pharmacokinetic response.

Aminolevulinic Acid↗

PubMed2000

Source of error in calculation of optical diffuse reflectance from turbid media using diffusion theory.

Diffusion theory and similarity relations were used to calculate the optical diffuse reflectance of an infinitely narrow laser beam incident upon a semi-infinite turbid medium. The results were analyzed by comparison with the accurate results from Monte Carlo simulations. Because a large number of photon packets were traced, the variance of the results from Monte Carlo simulations was small enough to reveal the detailed defects of the diffusion theory and the similarity relations, which are broadly used in photomedicine. We demonstrated that both diffusion theory and similarity relations provide very accurate results when the photon sources are isotropic and buried more deeply than one transport mean free path in turbid media. We found that the key factor affecting the accuracy of the diffusion theory application was the conversion from the infinitely narrow laser beam to an isotropic point source in turbid media.

Computer Simulation↗

PubMed2000

Modeling photon transport in transabdominal fetal oximetry.

The possibility of optical oximetry of the blood in the fetal brain measured across the maternal abdomen just prior to birth is under investigation. Such measurements could detect fetal distress prior to birth and aid in the clinical decision regarding Cesarean section. This paper uses a perturbation method to model photon transport through an 8-cm-diam fetal brain located at a constant 2.5 cm below a curved maternal abdominal surface with an air/tissue boundary. In the simulation, a near-infrared light source delivers light to the abdomen and a detector is positioned up to 10 cm from the source along the arc of the abdominal surface. The light transport [W/cm2 fluence rate per W incident power] collected at the 10 cm position is Tm = 2.2 x 10(-6) cm(-2) if the fetal brain has the same optical properties as the mother and Tf = 1.0 x 10(-6) cm(-2) for an optically perturbing fetal brain with typical brain optical properties. The perturbation P=(Tf - Tm)/Tm is -53% due to the fetal brain. The model illustrates the challenge and feasibility of transabdominal oximetry of the fetal brain.

Abdomen↗

PubMed1998

Crystallization and preliminary X-ray diffraction studies of homoserine dehydrogenase from Saccharomyces cerevisiae.

Recombinant homoserine dehydrogenase from Saccharomyces cerevisiae has been crystallized in three different forms. Crystals of the apo-enzyme belong to the tetragonal space group P4 and have unit-cell-dimensions a = b = 130 and c = 240 A. The resolution limit for these crystals is 3.9 A. Crystals of homoserine dehydrogenase grown in the presence of the co-factor NAD+ have the tetragonal space group P41212 or its enantiomorph P43212. The unit-cell dimensions for these crystals are a = b = 80.4 and c = 250.2 A, and the observed resolution limit is 2.2 A. Protein crystals grown in the presence of the product L-homoserine and the inert NAD+ analogue 3-aminopyridine adenine dinucleotide belong to the monoclinic space group P21 with unit-cell parameters a = 58.8, b = 104.2, c = 120.7 A, beta = 91.9 degrees. This last crystal form has a diffraction limit of 2.7 A resolution.

Crystallization↗

PubMed1998

Path integral description of light transport in tissue.

The early photons that arrive at a collector through a large thickness of tissue have potential for imaging internal organ structure, function, and status with improved image resolution relative to late arriving photons which have been diffusely scattered. Imaging algorithms require a theory to calculate early photon arrival for comparison with experimental data. The path integral description of light transport describes the movement of photons as particles undergoing collisions in a scattering medium based on the Brownian motion formalism of Feynman and Hibbs (unconstrained path) which applies the principle of least action. Including the additional constraint that photons have a constant velocity of c yields paths that conserve the speed of light (constrained path). This paper outlines the basic derivation of the path integral method and compares the constrained and unconstrained paths.

Animals↗

PubMed1998

Light distributions from point, line and plane sources for photochemical reactions and fluorescence in turbid biological tissues.

Light distributions in biological tissues are summarized in simple expressions for spherical, cylindrical and planar geometries due to point sources, line sources and planar sources. The goal is to provide workable tools for computing light distributions that govern the amount and distribution of photochemical reactions in experimental solutions, films and biological tissues. Diffusion theory expressions are compared with Monte Carlo simulations. Analytic expressions that mimic accurate Monte Carlo simulations are presented. Application to fluorescence measurements and prediction of necrotic zones in photodynamic therapy are outlined.

Animals↗

PubMed1997

CONV--convolution for responses to a finite diameter photon beam incident on multi-layered tissues.

A convolution program (CONV) solving responses to a collimated finite diameter photon beam perpendicularly incident on a multi-layered tissue has been coded in ANSI Standard C, hence, the program can be executed on various computers. The program, employing an extended trapezoidal rule for integration, convolves the responses to an infinitely narrow photon beam computed by a companion program (MCML). Dynamic data allocation is used for CONV as well as MCML, therefore, the number of tissue layers and grid elements of the grid system can be varied at run time. The potential error due to not scoring the first photon-tissue interactions separately is illustrated. The program, including the source code, has been in the public domain since 1992 and can be downloaded from the web site at http:(/)/biomed.tamu.edu/-lw.

Computer Simulation↗

PubMed1997

Perturbation theory for diffuse light transport in complex biological tissues.

A perturbation theory for the forward problem of optical transport in turbid media is developed. It is applicable to media with scattering and absorption in homogeneties and steady-state and modulated light. Absorbing perturbations can be described by a volume distribution of virtual sources that primarily causes a monopole perturbation light field. Scattering objects have an additional contribution that, in the limiting case of sharply bounded objects, is represented by a surface distribution of virtual sources and causes a dipolelike perturbation pattern. Using the concept of virtual sources, we discuss a possible ambiguity between the perturbations from scattering and absorbing inhomogeneities and the implications for the source-detector placement in inverse problems. We show that the surface effects due to sharp boundaries of scattering objects pose both a numerical problem and a chance to improve the resolution of inverse algorithms.

Absorption↗

PubMed1997

Diagnostic potential of laser-induced autofluorescence emission in brain tissue.

Laser-induced autofluorescence measurement of the brain was performed to assess its spectroscopic properties and to distinguish brain tumors from the normal tissues. The excitation-induced emission spectra were plotted on a 2-dimensional map, the excitation-emission matrix, to determine the excitation wavelengths most sensitive for the spectroscopic identification of brain tumors. The excitation-emission matrices of various types of human brain tumors and normal brain samples lead to the selection of three fluorescence peaks at 470, 520, and 630 nm, corresponding excitation light at 360, 440, and 490 nm, respectively for comparing the autofluorescence signatures of brain tissue. The fluorophores most likely related to each of these peaks are NAD(P)H, various flavins, and porphyrins, respectively. In vivo studies of rat gliomas showed that "NAD(P)H", "flavin", and "porphyrin" fluorescence were lower in gliomas than in normal brain. This finding suggests that there are certain relationship between brain tissue autofluorescence intensity and metabolic activity. In vitro human normal brain tissue fluorescence signals were lower in gray matter than in white matter and "NAD(P)H" fluorescence were lower in all measured human brain tumors than in normal brain. "Flavin" and "porphyrin" fluorescence in the neoplastic tissues was lower or higher than normal tissue depending on their nature. In conclusion, the fluorescence spectroscopic diagnostic system might be able to distinguish brain tumors from the normal brain tissue. The results of this study need to be verified and the investigation extended to human brain tumors in the operating room.

Brain↗

PubMed1996

Laser induced bubble formation in the retina.

BACKGROUND AND OBJECTIVE: The immediate thermodynamic effects of absorption of a laser pulse in the retina are theoretically studied to understand underlying physical damage mechanisms at threshold fluences. Damage is most likely to occur at threshold levels in the retinal pigment epithelium due to the strong absorption by the melanosomes. METHODS: The retinal pigment epithelium is modeled as an aqueous environment with absorption occurring at small spherical sites with absorption coefficients representative of melanosomes. For laser pulse durations of less than 10(-6) seconds, heat conduction is negligible during energy deposition and the resulting large energy density in the melanosomes will cause vaporization of the medium immediately surrounding a melanosome. RESULTS: We developed expressions for calculating the size of bubbles produced as a function of pulse characteristics and melanosome properties. We show that for pulse durations between 10(-6) and 10(-9) seconds, bubble formation will occur for laser fluences that are smaller than those required to cause Arrhenius-type thermal damage. CONCLUSION: Bubble formation is likely to be the mechanism of threshold damage to the retina for laser pulses durations in the time regime between 10(-6) and 10(-9) seconds.

Absorption↗

PubMed1996

Light transport in tissue: Accurate expressions for one-dimensional fluence rate and escape function based upon monte carlo simulation.

BACKGROUND AND OBJECTIVE: Surface laser irradiation of tissue often produces a fluence rate that varies only with depth. Modeling of laser-induced fluorescence involves an expression for the fraction of fluorescence emitted per unit depth that escapes from the medium. We present accurate expressions for fluence rate and escape function for the one-dimensional case based upon Monte Carlo simulation results. STUDY DESIGN/MATERIAL AND METHODS: Expressions were proposed for fluence rate, phi (z)/E0 = C1exp (-k1z/delta)-C2exp(-k2z/delta), and escape function, G(z) = C3exp(-k3z/delta), that varied solely with depth relative to effective penetration depth, z/delta. The scalar (C) and exponential (k) coefficient values were found by curve fitting the expressions to Monte Carlo simulation results. RESULTS: The coefficients varied as smooth functions of diffuse reflectance, Rd, for the range Rd = 0.01-0.8, and were independent of scattering anisotropy in the range g = 0.7-0.9. Simple expressions approximated the relationship of each coefficient to Rd. CONCLUSION: The proposed expressions have accuracy comparable to Monte Carlo simulations, over an essentially unrestricted range of diffuse reflectance values. The expressions may be combined accurately to portray laser-induced fluorescence measurements of a turbid medium.

Fluorescence↗

PubMed1996

Pulsed laser ablation of soft tissues, gels, and aqueous solutions at temperatures below 100 degrees C.

BACKGROUND AND OBJECTIVE: It is desirable for laser microsurgical procedures to remove tissue accurately and with minimal thermal and mechanical damage to adjacent non-irradiated tissues. Pulsed laser ablation can potentially remove biological tissue with microprecision if appropriate irradiation conditions are applied. The major goal of this study was to determine whether laser ablation is possible at temperatures below 100 degrees C. Another aim was to test thermoelastic and recoil stress magnitudes and to estimate their effects on phantom and biological tissue. STUDY DESIGN/MATERIALS AND METHODS: Pulsed laser ablation of water (aqueous solution of potassium chromate) and water containing soft tissues (collagen gel and pig liver) irradiated under confined stress conditions was studied. The ablation mechanism and stages of the ablation process were determined based on time-resolved measurements of laser-induced acoustic waves with simultaneous imaging of the ablation process by laser-flash photography. RESULTS: This study reveals the important role of tensile thermoelastic stress, which produces efficient cavitation that drives material ejection at temperatures substantially below 100 degrees C. Ablation thresholds for the aqueous solution, collagen gel, and liver were 20, 38, and 55 J/cm3, respectively, which correspond to temperature jumps of 5, 10, and 15 degrees C. Two distinct stages of material ejection were observed: (1) initial removal of small volumes of material due to the rupture of single subsurface bubbles, (2) bulk material ablation in the form of jets produced by intense hydrodynamic motions formed upon collapse of large bubbles after coalescence of smaller bubbles. The duration of material ejection upon short-pulse ablation may vary from microseconds to submilliseconds, and depended on the mechanical properties of materials and the incident laser fluence. CONCLUSION: Nanosecond laser ablation of water, gels, and soft tissue under confined-stress conditions of irradiation may occur at temperatures below 100 degrees C. This ablation regime minimizes thermal injury to adjacent tissues and involves thermoelastic stress and recoil pressure magnitudes, which may be tolerated by tissues adjacent to an ablated crater.

Animals↗